Plasma fractions for improved myelination

Abstract

Methods and compositions are described for restoring myelin levels in conditions associated with myelin degeneration, such as myelopathy associated with aging and age-related neurodegenerative and / or neuroinflammatory diseases or post-surgical recovery. The compositions used in the methods include plasma-derived plasma and plasma fractions that are effective in restoring myelin levels and / or improving neuronal conductance.

Description

[Technical Field]

[0001] The present invention relates to the prevention and treatment of certain age-related diseases. The present invention relates to the use of blood products, such as plasma and plasma fractions, to improve and accelerate recovery from surgery, including conditions and indications associated with surgery. The present invention further relates to the use of blood products, such as plasma and plasma fractions, to alleviate chronic pain or neurological disorders and treat indications associated with wound healing. The present invention relates to the use of blood products, such as plasma and plasma fractions, to treat and / or prevent age-related conditions, such as neurocognitive disorders and neurodegenerative disorders. Furthermore, the present invention relates to the use of blood products, such as plasma and plasma fractions, to restore myelin levels in conditions associated with myelin degeneration, for example, in aging and age-related neuroinflammatory diseases. Restoring myelin levels in such diseases improves nerve conductance. Accordingly, the present invention relates to methods for improving nerve myelination and / or nerve conductance by administering plasma or a plasma fraction to a subject with a condition associated with myelin degeneration. [Background technology]

[0002] The following is provided as background information and is not admitted to be prior art to the present invention.

[0003] Aging is an important risk factor for multiple human diseases, including cognitive impairment, cancer, arthritis, vision loss, osteoporosis, diabetes, cardiovascular disease, and stroke. In addition to the normal synapse loss during natural aging, synapse loss is an early pathological event common to many neurodegenerative conditions and is the primary correlate of the neurological and cognitive impairment associated with these conditions. Thus, aging remains the single most dominant risk factor for dementia-related neurodegenerative diseases such as Alzheimer's disease (AD) (Bishop, N.A. et al., Neural mechanisms of aging and cognitive decline. Nature 464(7288), 529-535(2010); Heeden, T. et al., Insights into the aging mind: a view from cognitive neuroscience. Nat. Rev. Neurosci. 5(2), 87-96(2004); Mattson, M.P. et al., Aging and neuronal vulnerability. Nat. Rev. Neurosci. 7(4), 278-294(2006)).

[0004] Aging affects all tissues and functions of the body, including the central nervous system, and declines in functions such as cognition can have a serious impact on quality of life. Treatments for cognitive decline and neurodegenerative disorders have had limited success in preventing and reversing impairment. Therefore, it is important to identify new therapies to maintain cognitive integrity by preventing, counteracting, or reversing the effects of aging.

[0005] Surgery is often associated with complications from pain, cardiopulmonary disease, infection, thromboembolic disease, and postoperative wound healing. Furthermore, wounds, whether from the surgery itself (e.g., incision) or from accident, violence, or illness followed by surgical intervention, require time to heal. Such complications are often exacerbated by age. The surgical stress response and subsequent demands on organ function can result in additional complications, often mediated by trauma-induced endocrine-metabolic changes and activation of cascades (cytokines, complement, arachidonic acid metabolites, nitric oxide, and free oxygen radicals) (Kehlet H., et al., Br. J. Anesthesia, 78:606-17 (1997)). During the surgical stress response, the sympathetic nervous system is activated (Starkweather A, Topics in Pain Management, 32(8): 1-11 (2017)) (Pinto PR, J Pain Res, 10:1087-98 (2017)). There is increased pituitary hormone secretion, leading to catabolic energy mobilization, which results in salt and water retention. Secretion of adrenocorticotropic hormone (ACTH) increases, and norepinephrine and sympathetic nervous activity increase. This leads to cardiovascular responses such as tachycardia and hypertension, and glucagon release, causing hyperglycemia. Increased growth hormone and cortisol inhibit the differentiation of monocytes into macrophages, which in turn inhibits T-cell signaling / histamine production and reduces immune cell migration (ibid.).

[0006] Current treatments for postoperative recovery include postoperative pain relief and multilevel interventions (ibid.). Pain management is important in many types of surgical recovery, and acute pain is expected (Pinto PR, J Pain Res, 10:1087-98 (2017)). Postoperative pain is associated with a significant proportion of patients undergoing general surgical procedures (Couceiro TC, Rev Bras Anestesiol, 59(3): 314-20 (2009)). Pain impairs healing and recovery, and therefore adversely affects clinical outcomes (ibid.). Hip and knee replacements are particularly associated with both chronic (e.g., osteoarthritis) and acute pain (ibid.). Therefore, analgesics are commonly used in postoperative recovery, both during the procedure and while patients recover at home.

[0007] Enhanced Recovery After Surgery (ERAS) is one type of multilevel intervention (Starkweather A, supra). ERAS focuses on a wide range of surgical procedures, including colorectal surgery, orthopedic surgery, gynecological surgery, urological surgery, head and neck cancer surgery, bladder cancer surgery, liver disease surgery, rectal / pelvic disease surgery, colon disease surgery, pancreaticoduodenectomy surgery, gastroenterectomy surgery, and bariatric and gynecological oncology surgery (ibid.). Multilevel strategies include enhancing preoperative techniques (counseling, fluid / carbohydrate administration, short-term fasting), perioperative techniques (short-acting anesthetics, normothermia, antibiotic prophylaxis, thromboembolic prophylaxis, salt / water overload prevention, vomiting prevention), and postoperative techniques (early oral feeding, exercise, nonopioid analgesics, and discharge assistance) (ibid.).

[0008] However, current therapies have not eliminated postoperative morbidity and mortality. Multi-step techniques, by their very nature, are time- and resource-consuming, and no single technology or drug treatment can match such a diverse regimen. Due to these shortcomings, new treatments are needed to improve postoperative recovery.

[0009] Degeneration of cerebral white matter is a key component of aging, as the ability to repair and replace healthy cells, which promote normal myelin regeneration processes, diminishes over time. Maintaining the integrity of the myelin sheath is important for ensuring proper axonal function and efficient signaling in neurons, and white matter loss contributes to cognitive impairment, particularly memory consolidation, in many neurodegenerative conditions. Due to myelin degeneration, there is a need for new therapies to restore myelin levels, improve nerve myelination, and / or improve nerve conductance in conditions associated with nerve myelin degeneration. Summary of the Invention

[0010] The present invention is based on the generation and use of blood products for the treatment and / or prevention of age-related disorders, such as cognitive impairment conditions, age-related dementia, and neurodegenerative diseases. The present invention recognizes, inter alia, the need for new therapies for the treatment and / or prevention of cognitive impairment, age-related dementia, and neurodegenerative diseases. The blood and plasma-derived compositions of the present invention address the failures and shortcomings of current therapies by utilizing plasma fractions that exhibit efficacy in the treatment and / or prevention of cognitive impairment, age-related dementia, and neurodegenerative diseases. Furthermore, the present invention relates to proteins identified in plasma fractions that may themselves exhibit efficacy as therapeutic or prophylactic agents for cognitive impairment and age-related dementia, or that are targets for investigation by additional agents.

[0011] The present invention is also based on the production and use of blood products to treat symptoms and conditions that affect surgical recovery, including, for example, pain and wound healing. The present invention recognizes, inter alia, the need for new therapies for the treatment of undesirable conditions associated with post-surgical recovery and the improvement of such recovery. The blood and plasma-derived compositions of the present invention address the failures and shortcomings of current therapies by utilizing plasma fractions that exhibit efficacy for treating undesirable conditions associated with post-surgical recovery and improving such recovery.

[0012] Furthermore, the present invention relates to the use of blood products such as plasma and plasma fractions to restore myelin levels in conditions associated with myelin degeneration, such as age-related neurocognitive and neurodegenerative disorders or myelopathy associated with post-surgical recovery. Restoring myelin levels in such diseases improves nerve conductance. Accordingly, certain embodiments of the present invention relate to methods of improving nerve myelination and / or improving nerve conductance by administering plasma or plasma fractions to a subject with a condition associated with myelin degeneration, such as, but not limited to, age-related neurocognitive and neurodegenerative disorders or myelopathy associated with post-surgical recovery.

[0013] The present invention is also based on the production and use of blood products to treat symptoms and conditions associated with acute and chronic pain. The present invention recognizes, among other things, the need for new therapies to alleviate pain. While therapies exist to treat acute and chronic pain, many of the therapies, such as opioid analgesics, exhibit high rates of addiction, abuse, and associated morbidity and mortality.

[0014] The present invention also recognizes that differences in protein content between different plasma fractions (e.g., fractions, effluents, "plasma fractions," plasma protein fractions, human albumin solutions) may contribute to the prevention and / or amelioration of certain cognitive impairments and alleviation of neurodegenerative diseases. By way of example and not limitation, embodiments of the present invention demonstrate that simply the high albumin concentration of Human Albumin Solution (HAS) preparations is not the driving force behind the cognitive improvements associated with Plasma Protein Fraction (PPF) preparations with lower albumin concentrations.

[0015] Blood and plasma from young donors have shown improvement and reversal of existing effects of brain aging, including at molecular, structural, functional, and cognitive levels (Saul A. Villeda, et al. Young blood reverses age-related impairments in cognitive function and synaptic plasticity in mice. Nature Medicine 20 659-663 (2014)). The present invention relates to plasma fractions and effluents, some of which are traditionally used to treat shock in patients, and the discovery that plasma fractions and effluents are effective as treatments for age-associated cognitive impairment.

[0016] Thus, according to aspects of the present invention, there are provided methods for treating age-related cognitive impairment, age-related dementia, and / or neurodegenerative diseases using a blood product fraction of plasma. Aspects of the method include administering the plasma fraction to an individual suffering from or at risk of developing age-related cognitive impairment or a neurodegenerative disease. Further aspects of the method include administering a plasma fraction derived from a pool of donors within a specific age range to an individual suffering from or at risk of developing age-related cognitive impairment. Reagents, devices, and kits useful for practicing the method are also provided.

[0017] In one embodiment, the plasma fraction can be one of several plasma fractions obtained from a blood fractionation process, such as the Cohn fractionation process described below. In another embodiment, the plasma fraction is a solution containing normal human albumin, alpha globulins, beta globulins, gamma globulins, and other proteins, either individually or in complexes. The plasma fraction may be of the type referred to herein as a "plasma fraction." In another embodiment, the plasma fraction may be of the type known to those skilled in the art as a "plasma protein fraction" (PPF). In another embodiment, the plasma fraction may be a "human albumin in solution" (HAS) fraction. In yet another embodiment, the plasma fraction may have substantially all of the clotting factors removed so as to preserve the efficacy of the fraction with a reduced risk of thrombosis. Embodiments of the invention may also include administering fractions derived from, for example, a young donor or a young donor pool. Another embodiment of the invention may include monitoring cognitive improvement in subjects treated with the plasma fraction.

[0018] All publications and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference. [Brief explanation of the drawings]

[0019] The accompanying drawings illustrate embodiments of the invention and, together with this description, serve to explain the invention. These drawings are offered by way of example and not by way of limitation. It is emphasized that various features of the drawings may not be to scale.

[0020] [Figure 1] Rearing latency of 3- or 13-month-old NSG mice treated with control, PPF1, or HAS1 is shown when placed in an open field chamber for 15 minutes. [Figure 2] Shown is the locomotor speed of 3- or 13-month-old NSG mice treated with control, PPF1, or HAS1 placed in an open field chamber for 15 minutes. [Figure 3] Shown is the distance traveled by 3- or 13-month-old NSG mice treated with control, PPF1, or HAS1 when placed in an open field chamber for 15 minutes. [Figure 4]1 shows the time spent in the novel arm in a cued Y-maze test by 3- or 13-month-old NSG mice treated with control, PPF1, or HAS1. [Figure 5] Figure 1 shows the ratio of time spent in the novel arm to the familiar arm (novel:familiar ratio) in 3- or 13-month-old NSG mice treated with control, PPF1, or HAS1 in a cued Y-maze test. [Figure 6] 1 shows the movement speed in a cued Y-maze test of 3- or 13-month-old NSG mice treated with control, PPF1, or HAS1. [Figure 7] 1 shows the distance traveled in a cued Y-maze test by 3- or 13-month-old NSG mice treated with control, PPF1, or HAS1. [Figure 8] (A) Percentage of freezing time in a contextual fear conditioning test of memory for 3- and 13-month-old NSG mice treated with control, PPF1, or HAS1. (B) Percentage of freezing time in an auditory cued fear conditioning test of memory for 3- and 13-month-old NSG mice treated with control, PPF1, or HAS1. [Figure 9] Quantification of the percent freezing time during the last 90 seconds of a cued fear conditioning test for memory in 3- and 13-month-old NSG mice treated with control, PPF1, or HAS1. [Figure 10] (A) Chart of Barnes maze latency, testing spatial memory. The latency for 3- and 13-month-old NSG mice treated with control, PPF1, or HAS1 to reach the target hole is shown. (B) Quantification of the average of the last three trials shown in (A). [Figure 11]A, quantification of the number of cells staining positive for doublecortin (Dcx), a marker of newborn neurons, in the dentate gyrus of 3- and 13-month-old NSG mice treated twice weekly with control, PPF1, or HAS1 for up to 6 months. B, quantification of the number of cells staining positive for Ki67, a marker of proliferating cells, in the dentate gyrus of 3- and 13-month-old NSG mice treated twice weekly with control, PPF1, or HAS1 for up to 6 months. [Figure 12] The number of cells staining positive for Dcx was quantified in 13-month-old NSG mice treated with control, PPF1, 1x concentrated HAS1, or 5x concentrated HAS1 three times a week for 5 weeks. [Figure 13] The number of cells staining positive for Ki67 was quantified in 13-month-old NSG mice treated with control, PPF1, 1x concentrated HAS1, or 5x concentrated HAS1 three times a week for 5 weeks. [Figure 14] (A) shows the number of rearings in the open field chamber of NODscid mice treated intravenously with either saline (control) or PPF1 via tail vein injection twice a week starting at 6 months of age. Rearing was measured over a 15-minute span after placing the mice in the open field chamber. (B) shows the locomotor speed in the open field chamber of mice treated intravenously with either saline (control) or PPF1 via tail vein injection twice a week starting at 6 months of age. Speed ​​was measured over a 15-minute span after placing the mice in the open field chamber. (C) shows the distance traveled in the open field chamber of mice treated intravenously with either saline (control) or PPF1 via tail vein injection twice a week starting at 6 months of age. Speed ​​was measured over a 15-minute span after placing the mice in the open field chamber. [Figure 15]Barnes maze latency and hippocampus-dependent spatial learning and memory are shown. The latency to reach the target hole is shown for aged NSG mice (12 months old) treated with 150 μL of saline control, young plasma, effluent I, or effluent II / III. [Figure 16] Figure 1 shows the effects of young human plasma, PPF1, and saline control on hippocampus-dependent spatial learning and memory in male aged NSG mice (12 months old). Mice were treated with 150 μL of purified young human plasma (young plasma), PPF1, or saline three times per week (iv) for four weeks, and twice per week during weeks 5 and 6, the weeks in which the reported tests were performed. The latency to reach the hole in the Barnes maze is shown for each treatment group. [Figure 17] Figure 1 shows the effects of young human plasma, PPF1, and saline control on the mean latency to find the target hole in the Barnes maze during the last three trials of each test day. Aged NSG mice (12 months old) were treated with 150 μL of purified young human plasma (young plasma), PPF1, or saline three times (iv) per week for 4 weeks, followed by twice per week during weeks 5 and 6, the weeks in which testing was performed. [Figure 18] The effects of young human plasma, PPF1, and saline control on cell viability as determined by BrdU detection are shown. Aged NSG mice (12 months old) were treated with 150 μL of purified young human plasma (young plasma), PPF1, or saline three times per week (iv) for 4 weeks, followed by treatment twice per week during weeks 5 and 6, the weeks when behavioral testing was performed. Hippocampal slices were analyzed after sacrifice. [Figure 19] 1 shows the effect of control, PPF1, and HAS1 on the proliferation of neurospheres in cortical cultures. Shown are example images of neurospheres in cortical cultures after 21 days of in vitro culture, imaged with Tuj1, DAPI, or Tuj1 and DAPI. [Figure 20] 1 shows the effects of control, PPF1, and HAS1 on net neurite length in cortical cultures. [Figure 21]The effects of vehicle, PPF1, and HAS1 on sphere and process growth in cortical cultures are shown. Yellow shading highlights spheres and pink shading highlights neurites as determined by the IncuCyte software algorithm (Essen BioScience, Inc., Ann Arbor, MI). [Figure 22] A shows the number of neurospheres quantified as a percentage of vehicle in the cortex of E14-15 mouse embryos suspended in Neurobasal Medium supplemented with B27 and 2 mM Glutamax (vehicle), PPF1 (10% of a 5% stock solution), or HAS1 (10% of a 5% stock solution).B shows the neurite length quantified as a percentage of vehicle in the cortex of E14-15 mouse embryos suspended in Neurobasal Medium supplemented with B27 and 2 mM Glutamax (vehicle), PPF1 (10% of a 5% stock solution), or HAS1 (10% of a 5% stock solution). (C) Neurite branch points were quantified as a percentage of vehicle in the cortex of E14-15 mouse embryos suspended in Neurobasal medium supplemented with B27 and 2 mM Glutamax (vehicle), PPF1 (10% of a 5% stock solution), or HAS1 (10% of a 5% stock solution). (D) Neurosphere size was quantified as a percentage of vehicle in the cortex of E14-15 mouse embryos suspended in Neurobasal medium supplemented with B27 and 2 mM Glutamax (vehicle), PPF1 (10% of a 5% stock solution), or HAS1 (10% of a 5% stock solution). [Figure 23] Quantification of the number of neurospheres staining positive for Sox2 treated with control vehicle (Neurobasal medium supplemented with B27 and 2 mM Glutamax), PPF1 (10% of a 5% stock solution), or HAS1 (10% of a 5% stock solution) is shown. Sox2 staining is an indicator of the neurogenic potential of the neurospheres. [Figure 24]Chronic constriction injury (CCI) experiments were performed on 23-month-old wild-type mice. 24 hours before CCI surgery or sham surgery was performed, with administration of either PPF1, gabapentin, recombinant human albumin (rhAlb), or vehicle control in a 7-day continuous pulse-dose regimen. Behavior was assessed from week 2 to week 5, and tissue collection for histological examination was performed at week 5. [Figure 25] Figure 1 shows the location of CCI surgery performed on a 22-month-old wild-type mouse. Ligatures were placed on the sciatic nerve as shown. This figure is adapted from Suter MR, et al., Anesthesiology Res and Practice (2011), which is incorporated herein by reference in its entirety. [Figure 26] Figure 24 shows data from the von Frey mechanical allodynia test using wild-type mice treated with CCI or sham surgery. Von Frey filament stimulation of the weakened hind paw via the sciatic nerve was useful for analyzing pain-related behaviors. The pressure with which the mice withdrew their hind paw was measured and plotted. This figure shows that mice treated with PPF1 after CCI surgery were significantly less painful (able to tolerate greater pressure) than mice treated with vehicle control after CCI surgery. Mice treated with vehicle after sham surgery also showed significantly less pain than mice treated with vehicle control after CCI surgery. This indicates that PPF1 has a positive effect on mechanical nociceptive deficits. [Figure 27] Figure 2 shows data from histological examination of the hippocampus performed on wild-type mice described in Figure 24. Neurogenesis was measured using the doublecortin (DCX) marker. Mice treated with PPF1 after CCI surgery had significantly more hippocampal neurogenesis than mice treated with vehicle after CCI surgery. Mice that underwent sham surgery and were treated with vehicle tended to have increased neurogenesis compared to mice treated with vehicle after CCI surgery. Thus, PPF1 demonstrated the ability to restore neurogenesis after chronic nerve injury. [Figure 28]Figure 24 shows data from histological examination of the hippocampus performed on wild-type mice described in Figure 24. CD68 expression was quantified, and it was shown that mice that underwent CCI surgery and vehicle administration had significantly higher numbers of CD68-positive cells in the hippocampus than mice that underwent CCI surgery and PPF1 administration. Similar differences were observed between mice that underwent CCI surgery and vehicle administration and mice that underwent sham surgery and vehicle administration. This indicates that PPF1 helps suppress neuroinflammation caused by chronic nerve injury. [Figure 29] Figure 2 shows data from the von Frey mechanical allodynia test performed on 22-month-old C57BL / 6J mice undergoing CCI surgery or sham surgery and tested according to the timeline shown in Figure 24. The pressure at which the mice withdrew their hind paws was assessed and is expressed as the number of weeks after CCI or sham surgery. The data show that mice treated with PPF1 after CCI surgery had significantly increased tolerance to mechanical nociception at all time points assessed compared with mice treated with vehicle after CCI surgery. Conversely, mice treated with gabapentin only showed significant improvement in mechanical nociception at 2 weeks after CCI surgery, remaining similar to vehicle-treated mice at other time points. Sham-operated mice showed significantly increased responses to mechanical nociception at 3 and 5 weeks after surgery. Together, these data demonstrate that PPF1 improves peripheral pain for a longer period than standard treatment (gabapentin). [Figure 30]Figure 2 shows data from the hot plate test performed on 22-month-old wild-type mice that underwent CCI surgery or sham surgery and were tested according to the timeline described in Figure 24. The assay was performed as described by Woolfe and Macdonald (Woolfe G. and Macdonald AD, J. Pharmacol. Exp. Ther. 80:300-07 (1944), incorporated herein by reference in its entirety). The hot plate was set to a temperature of 55°C. Mice were placed in the transparent cylinder for 30 minutes to allow for acclimation. The cylinder was placed on the hot plate and a timer was started. The first observation of nocifensive behavior (e.g., hind paw licking, jumping) was recorded as the duration. Hot plate nocifensive duration is shown 5 weeks after CCI surgery or sham surgery. PPF1-treated mice were significantly less sensitive to hot plate stimulation than mice given CCI surgery and vehicle control, demonstrating the rescue effect of PPF1. [Figure 31] 24 shows data from hot plate testing performed on wild-type mice that underwent CCI surgery or sham surgery and were tested according to the timeline shown in FIG. 24. Hot plate nocifensive duration is shown 5 weeks after CCI surgery or sham surgery. PPF1- and rhAlb-treated mice were significantly less sensitive to hot plate stimulation than mice that received CCI surgery plus vehicle control. [Figure 32] Figure 2 shows data from von Frey mechanical allodynia testing performed on C57BL / 6J mice that underwent CCI or sham surgery and were tested according to the timeline shown in Figure 24. Mice treated with PPF1 after CCI surgery exhibited significantly increased mechanical nociception at all time points assessed compared to vehicle-treated mice after CCI surgery. Conversely, mice treated with rhAlb exhibited a similar response to mechanical allodynia as vehicle-treated mice at all time points. [Figure 33]Figure 24 shows data from sciatic nerve histological analysis (approximately 1000 μm distal to the last node) of myelin basic protein (MBP) expression in C57BL / 6J mice that underwent CCI or sham surgery and were analyzed after tissue collection 35 days later. As shown in Figure 24, mice administered PPF1 after CCI surgery and analyzed after tissue collection 35 days later showed significantly increased MBP intensity, suggesting increased myelin expression compared to vehicle-treated animals. Sham-operated mice also showed increased MBP compared to CCI-injured mice. [Figure 34] Figure 24 shows data from sciatic nerve histological analysis (approximately 1000 μm distal to the last node) of S-100 protein (expressed by Schwann cells) in C57BL / 6J mice that underwent CCI or sham surgery and were analyzed after tissue collection 35 days later. The data show that mice treated with PPF1 after CCI surgery had significantly increased S-100 intensity compared to vehicle-treated animals, suggesting an increase in Schwann cells (cells that produce myelin in peripheral nerves). Sham-operated mice also had increased S-100 intensity compared to vehicle-treated mice after CCI injury. [Figure 35] Selected images from histological analysis of sciatic nerves identifying the location (approximately 1000 μm distal to the last node) used for quantification in Figures 33 and 34, and representative intensities of S-100 protein (indicated in Schwann cells) and myelin basic protein in C57BL / 6J mice that underwent CCI surgery as shown in Figure 24, were treated with either vehicle or PPF1, and were used for quantitative analysis of sciatic nerves from day 35 onwards. [Figure 36]Figure 24 shows spinal cord histological analysis data (performed on spinal cord tissue collected from lumbar regions L4-L6) from C57BL / 6J mice that underwent CCI or sham surgery and were analyzed 35 days after tissue collection. The data show that mice treated with PPF1 after CCI surgery had significantly reduced BDNF levels in the dorsal horn of the spinal cord, suggesting reduced microglial activity in the spinal cord. Because BDNF is a pro-inflammatory cytokine released by activated microglia, these findings suggest that PPF1 reduces a fundamental regulator of pain states in the spinal cord, normalizing its levels to those of sham-operated (non-CCI) mice. [Figure 37] Figure 24 shows spinal cord histological analysis data (performed on spinal cord tissue collected from lumbar regions L4-L6) from C57BL / 6J mice that underwent CCI or sham surgery and were analyzed 35 days after tissue collection. The data show that mice treated with PPF1 after CCI surgery had significantly reduced CD68 intensity within the dorsal horn of the spinal cord, suggesting reduced microglial activity within the spinal cord. Because CD68 protein is expressed by activated microglia, PPF1 reduces the activity of a fundamental cell type involved in inducing pain states within the spinal cord, normalizing it to the level of sham-operated (non-CCI-injured) mice. The data shown in Figures 36 and 37 indicate that PPF1 centrally regulates pain states resulting from sciatic nerve injury, improving or preventing pain signaling between peripheral nerves and the brain, also known as central sensitization. [Figure 38] Selected images from histological analysis of the spinal cord identifying the location of the spinal cord horns used for quantification in Figure 37 (performed on spinal cord tissue collected from lumbar regions L4-L6) and representative intensity of CD68 protein (expressed by activated microglia) from C57BL / 6J mice that underwent CCI surgery and were treated with either vehicle or PPF1, as shown in Figure 24, and used for quantitative analysis of spinal cord tissue from day 35 onwards. [Figure 39]Selected images from histological analysis of the spinal cord identifying the location of the spinal cord horns used for quantification in Figure 36 (performed on spinal cord tissue collected from lumbar regions L4-L6) and representative intensity of BDNF protein (released by activated microglia) from C57BL / 6J mice that underwent CCI surgery and were treated with either vehicle or PPF1, as shown in Figure 24, and used for quantitative analysis of spinal cord tissue from day 35 onwards. [Figure 40] Chronic constriction injury (CCI) experiments were performed on 22-month-old wild-type mice. Two weeks before administration of either PPF1, rhAlb, or vehicle control in a 7-day continuous pulse administration regimen, CCI surgery or sham surgery were performed by ligation. Behavior was assessed weekly from week 2 to week 7, and tissue collection for histological examination was performed at week 7. [Figure 41] Figure 4 shows data from the von Frey mechanical allodynia test using C57BL / 6J mice that underwent CCI or sham surgery and were tested according to the timeline shown in Figure 40. Mice treated with PPF1 2 weeks after CCI surgery showed a significant increase in tolerance to mechanical nociception beginning 1 week after cessation of PPF1 treatment, which was maintained throughout the test. This finding suggests that PPF1 treatment initiates a process of longitudinally decreasing sensitivity to mechanical allodynia, with improved tolerance not evident until 1 week after treatment (in contrast to treatments such as opioid analgesics, which only provide benefit during treatment) and persisting for at least 28 days. Conversely, mice treated with rhAlb exhibit a similar response to mechanical allodynia as vehicle-treated mice at all time points. [Figure 42] 40. Data from the hot plate test performed on wild-type mice that underwent CCI surgery or sham surgery and were tested according to the timeline described in FIG. 40. The duration of hot plate nocifensive responses 5 weeks after CCI surgery or sham surgery is shown. PPF1-treated mice are significantly less sensitive to hot plate stimulation than mice that received CCI surgery plus vehicle control. [Figure 43]40. Data from the hot plate test performed on wild-type mice that underwent CCI surgery or sham surgery and were tested according to the timeline described in FIG. 40. The duration of hot plate nocifensive responses 7 weeks after CCI surgery or sham surgery is shown. PPF1-treated mice are significantly less sensitive to hot plate stimulation than mice that received CCI surgery plus vehicle control. [Figure 44A] Figure 1 shows that plasma fraction treatment reduces neuroinflammation and enhances neurogenesis, and shows a schematic diagram of the plasma fractionation process. Data shown are mean ± SEM, *p<0.05, **p<0.01, ***p<0.001, Veh=vehicle. [Figure 44B] A schematic diagram of the study design shows that plasma fraction treatment reduces neuroinflammation and enhances neurogenesis. Wild-type male mice aged 22 to 24 months were treated with PPF1 and analyzed 10 days (CD68 / Iba-1) or 6 weeks (BrdU / DCX). Data shown are mean ± SEM; *p<0.05, **p<0.01, ***p<0.001; Veh=vehicle). [Figure 44C] Plasma fraction treatment reduced neuroinflammation and enhanced neurogenesis. Quantification of CD68 and Iba-1 immunoreactivity in the hippocampus suggests that PPF1 treatment reduced microglia. Data shown are mean ± SEM; *p<0.05, **p<0.01, ***p<0.001; Veh=vehicle. [Figure 44D] Figure 1 shows that plasma fraction treatment reduces neuroinflammation and enhances neurogenesis. Quantification of BrdU and DCX immunoreactivity in the hippocampus suggests that PPF1 treatment improved cell survival and neurogenesis. Data shown are mean ± SEM; *p<0.05, **p<0.01, ***p<0.001; Veh=vehicle. [Figure 45A]Figure 1 shows a representative image of the hippocampus from an 11-month-old mouse, illustrating age-related myelin loss in the hippocampus. The box on the right highlights the CA1 ROI indicated in the image. Scale bars = 500 μm, 100 μm, and 20 μm. [Figure 45B] Figure 1 shows a representative image of the hippocampus from a 24-month-old mouse, illustrating age-related myelin loss in the hippocampus. The box on the right highlights the CA1 ROI indicated in the image. Scale bars = 500 μm, 100 μm, and 20 μm. [Figure 45C] Figure 1 shows age-related myelin loss in the hippocampus, demonstrating that myelin coverage in the hippocampus and cortex remains unchanged between 11-month-old and 24-month-old mice. Data shown are mean ± SEM, Mann-Whitney test, **p<0.003. [Figure 45D] Figure 1 shows age-related myelin loss in the hippocampus, with the mean optical density of MBP signal significantly increased in the hippocampus and CA1 in 11-month-old mice compared to 24-month-old mice. Data shown are mean ± SEM, Mann-Whitney test, **p<0.003. [Figure 45E] Representative images of PDGFRa+ cells in the hippocampus of 11-month-old and 24-month-old mice showing age-related myelin loss in the hippocampus. Scale bars = 500 μm, 100 μm, and 20 μm. [Figure 45F] Figure 1 shows age-related myelin loss in the hippocampus, and demonstrates that the density of PDGFRa+ cells in the hippocampus did not change with age. Data shown are mean ± SEM, Mann-Whitney test, **p<0.003. [Figure 46A] We demonstrate that the Hhcy and cisplatin models do not exhibit a deficiency in myelin content, and present a protocol for inducing Hhcy in 12-week-old mice using a folate-deficient diet for 10 weeks. [Figure 46B]This demonstrates that the AHhcy and cisplatin models do not exhibit a deficit in myelin content; no differences were observed in myelin coverage, hippocampal MBP optical density, or OPC density as measured by hippocampal PDGFRa. All data shown are means ± SEM. [Figure 46C] Illustrating that the Hhcy and cisplatin models do not exhibit a deficiency in myelin content, a schematic diagram of the protocol for inducing cognitive impairment in 7-month-old mice by IP administration of 2.3 mg / kg cisplatin is shown. [Figure 46D] This demonstrates that the Hhcy and cisplatin models do not exhibit a deficit in myelin content, with no differences observed in myelin coverage, hippocampal optical density, or OPC density as measured by hippocampal PDGFRa. All data shown are means ± SEM. [Figure 47A] FIG. 1 shows that myelin content in the hippocampus and cortex is increased in aged mice treated with PPF1, and a schematic diagram of the experimental protocol is shown. [Figure 47B] Aged mice treated with PPF1 show increased myelin content in the hippocampus and cortex. Representative images of the hippocampal ROI (inset) and dentate gyrus show increased MBP expression in PPF1-treated mice. Scale bar = 200 μm. [Figure 47C] The results show that myelin content in the hippocampus and cortex was increased in aged mice treated with PPF1, and that myelin coverage and MBP optical density in the hippocampus and CA1 were increased in PPF1-treated mice (Spearman correlation test, R = 0.7182, *p = 0.0162; Mann-Whitney test, ****p < 0.0001, ***p < 0.0002, *p = 0.03). [Figure 47D] Representative images of MBP expression in the cortex (inner dotted line) and ROI (outer dotted line) show increased myelin content in the hippocampus and cortex in aged mice treated with PPF1. Scale bar = 200 μm. [Figure 47E]This shows that aged mice treated with PPF1 have increased myelin content in the hippocampus and cortex, and increased MBP expression is observed in the cortex of PPF1-treated mice (Spearman correlation test, R=0.7182, *p=0.0162; Mann-Whitney test, ****p<0.0001, ***p<0.0002, *p=0.03). [Figure 47F] Aged mice treated with PPF1 showed increased myelin content in the hippocampus and cortex, but no difference in PDGFRa+ OPC density was observed in the hippocampus (Spearman correlation test, R=0.7182, *p=0.0162; Mann-Whitney test, ****p<0.0001, ***p<0.0002, *p=0.03). [Figure 47G] Aged mice treated with PPF1 showed increased myelin content in the hippocampus and cortex, and a significant correlation was observed between MBP expression and Y-maze performance (percentage of time in the novel arm) in PPF1-treated mice. DETAILED DESCRIPTION OF THE INVENTION

[0021] 1. Introduction The present invention relates to the identification and discovery of methods and compositions for the treatment and / or prevention of cognitive impairment, including age-related dementia and neurodegenerative diseases. Described herein are methods and compositions for treating subjects suffering from such disorders, which are aspects of the present invention. The methods and compositions described herein are useful for preventing cognitive impairment, age-related dementia, and neurodegenerative diseases; alleviating symptoms of cognitive impairment, age-related dementia, and neurodegenerative diseases; slowing the progression of age-related cognitive impairment, age-related dementia, and neurodegenerative diseases; and / or reversing the progression of age-related cognitive impairment, age-related dementia, and neurodegenerative diseases.

[0022] The present invention also relates to the identification and discovery of methods and compositions for treating undesirable conditions associated with post-surgical recovery and improving such recovery. By "improving such recovery," it is meant that a subject's post-surgical recovery may be accelerated, i.e., the subject may be able to be transferred or discharged from inpatient care in a shorter time than would be the case without the intervention of embodiments of the present invention. "Undesirable condition" refers to conditions and symptoms such as, by way of example and not limitation, pain, cardiopulmonary disease, infection, thromboembolic disease, inflammation, and delayed wound healing. Described herein are aspects of the present invention, methods and compositions for treating and improving a subject suffering from an undesirable condition associated with post-surgical recovery, and methods and compositions for improving such recovery. Also described herein are dosing regimens for inducing improvement and improving recovery in a subject suffering from an undesirable condition associated with post-surgical recovery. The methods and compositions described herein are useful for preventing complications from post-surgical recovery, ameliorating symptoms of preventing complications from post-surgical recovery, and promoting post-surgical recovery. The methods and compositions of the present invention may be utilized or administered preoperatively (before surgery), perioperatively (during surgery), or post-operatively (after surgery).

[0023] Another aspect of the present invention is directed to treating chronic pain / neuropathy more generally, and not solely to treating chronic pain / neuropathy associated with post-surgical recovery. The methods and compositions described herein may be used to treat chronic pain and neuropathy. By "treating chronic pain and neuropathy," we mean a slight, moderate, or significant reduction in the level of chronic pain experienced by a subject administered a composition of the present invention, as assessed by subjective or objective means. Such means may include, by way of example and not limitation, self- or medical professional-administered tests such as x-rays, MRIs, CT scans, patient pain ratings or descriptions, range of motion, reflexes, muscle strength, sensitivity (e.g., the time it takes a subject to remove a limb from pressure or other stimulation), blood tests for inflammatory markers, electromyography (EMG), and nerve conduction velocity.

[0024] A further aspect of the present invention is the use of blood products such as plasma and plasma fractions to restore myelin levels in conditions associated with myelin degeneration, such as age-related neurocognitive and neurodegenerative disorders or myelopathy associated with post-surgical recovery, which results in improved nerve conductance.

[0025] Accordingly, certain embodiments of the present invention provide methods for restoring myelin levels and / or improving neural conductance, comprising administering an effective amount of a plasma fraction to a subject diagnosed with a condition associated with myelin degeneration. The condition associated with myelin degeneration may be a neurodegenerative and / or neuroinflammatory condition, such as an age-related neurocognitive condition, a neurodegenerative condition, or a neuroinflammatory condition. In some cases, the condition associated with myelin degeneration is a myelopathy associated with post-surgical recovery.

[0026] In some cases, the plasma fraction is a plasma protein fraction (PPF), which may be commercially available. PPF has a total protein content of at least 83% but less than 95% albumin and 17% or less globulin. PPF also has 1% or less gamma globulin.

[0027] The plasma fraction may be derived from plasma obtained from a pool of humans, for example, aged 0 to 40 years (e.g., 0, 1, 5, 10, 15, 20, 25, 30, 35, 40 years).

[0028] Plasma fractions can be prepared from mammalian blood products, particularly human blood products.

[0029] The subject may be a mammal, particularly a human.

[0030] One embodiment of the present invention involves the use of a plasma fraction, such as one or more fractions or effluents obtained from a blood fractionation process, such as the Cohn fractionation process described below, as a treatment. One embodiment of the present invention involves the use of a plasma fraction (a solution containing normal human albumin, alpha and beta globulins, gamma globulins, and other proteins, individually or in complexes, hereafter referred to as the "plasma fraction"). Another embodiment of the present invention involves the use of a plasma protein fraction (PPF) as a treatment. Another embodiment of the present invention involves the use of a human albumin solution (HAS) fraction as a treatment. Yet another embodiment involves the use of an effluent from a blood fractionation process, such as Effluent I or Effluent II / III described below. Further embodiments include plasma fractions from which substantially all coagulation factors have been removed to reduce the risk of thrombosis while retaining efficacy (see, e.g., U.S. Patent Application Nos. 62 / 236,710 and 63 / 376,529, which are incorporated herein by reference in their entireties).

[0031] Before describing the present invention in detail, it is to be understood that this invention is not limited to the particular methods or compositions described, as these may, of course, vary. Also, it is to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only by the appended claims.

[0032] The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein should be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention. Further, the dates of publication provided may be different from the actual publication dates, which may need to be independently confirmed.

[0033] When a range of values ​​is provided, each intervening value between the upper and lower limit of that range is also understood to be specifically disclosed, to the tenth of the unit of the lower limit, unless the context clearly indicates otherwise. Each smaller range between any stated or intervening value in a stated range and any other stated or intervening value in that stated range is encompassed within the invention. The upper and lower limits of such smaller ranges may independently be included or excluded from the range, and each range where either or both limits are included in such smaller ranges, or where neither limit is included in such smaller ranges, is also encompassed within the invention, unless there is a specifically excluded limit in the stated range. When a stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the invention.

[0034] However, the claims may be drafted to exclude any optional element, and therefore, this statement is intended to serve as a basis precedent for using exclusive terminology such as "solely," "only," and the like, or for using a "negative" limitation in connection with the recitation of claim elements.

[0035] As will be apparent to those skilled in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has individual components and features that may be readily separated from or combined with the features of any of the other several embodiments without departing from the scope and spirit of the invention. Any recited method may be carried out in the order of events recited or in any other order that is logically possible.

[0036] 2.Definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, some potentially preferred methods and materials are now described. All publications mentioned herein are incorporated by reference to disclose and describe the methods and / or materials in connection with the cited publications. In case of conflict, it is understood that the present disclosure supersedes any disclosure content of the incorporated publication.

[0037] It should be noted that, as used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a cell" includes a plural of such cells, reference to "the peptide" includes reference to one or more peptides and equivalents thereof known to those skilled in the art, e.g., polypeptides, and so forth.

[0038] In describing the methods of the present invention, the terms "host," "subject," "individual," and "patient" are used interchangeably and refer to any mammal in need of such treatment according to the methods of the present disclosure. Such mammals include, for example, humans, sheep, cows, horses, pigs, dogs, cats, non-human primates, mice, and rats. In certain embodiments, the subject is a non-human mammal. In some embodiments, the subject is a livestock animal. In other embodiments, the subject is a pet. In some embodiments, the subject is a mammal. In certain instances, the subject is a human. Other subjects can include domestic pets (e.g., dogs and cats), livestock (e.g., cows, pigs, goats, horses, etc.), rodents (e.g., mice, guinea pigs, and rats, e.g., in animal models of disease), and non-human primates (e.g., chimpanzees and monkeys). Thus, subjects of the present invention include, but are not limited to, mammals, such as humans and other primates, such as chimpanzees and other apes and monkey species; in certain embodiments, the subject is a human. The term subject is also intended to include an individual or organism of any age, weight, or other physical characteristic, and the subject may be an adult, child, infant, or newborn.

[0039] "Young" or "young individual" refers to an individual who is 40 years of age or younger, e.g., 35 years of age or younger, e.g., 30 years of age or younger, e.g., 25 years of age or younger, or 22 years of age or younger. In some cases, the individual from whom the blood product, including young plasma, is derived is 10 years of age or younger, e.g., 5 years of age or younger, e.g., 1 year of age or younger. In some cases, when a plasma product is collected from the umbilical cord of a newborn, the subject is a newborn and the source of the plasma product is the umbilical cord. Thus, "young" and "young individual" can refer to a subject aged 0-40 years, e.g., 0, 1, 5, 10, 15, 20, 25, 30, 35, or 40 years. In other cases, "young" and "young individual" can refer to biological age (as opposed to chronological age), such as an individual who does not exhibit levels of inflammatory cytokines in plasma exhibited in older individuals. Conversely, these "young" and "young individual" may refer to biological age (as opposed to chronological age), such as an individual exhibiting a high level of anti-inflammatory cytokines in plasma compared to the levels in relatively older individuals. By way of example and not limitation, the inflammatory cytokine is eotaxin, and the fold difference between young subjects or young individuals and older individuals is at least 1.5-fold. Similarly, the fold difference of other inflammatory cytokines between older and younger individuals may be used to refer to biological age (see U.S. Patent Application No. 13 / 575,437, incorporated herein by reference). Typically, the individual is healthy. For example, the individual does not have a hematological malignancy or autoimmune disease at the time of collection.

[0040] "An individual suffering from or at risk of suffering from age-related cognitive impairment" refers to an individual who has passed more than about 50% of their life expectancy, for example, more than 60%, for example, more than 70%, for example, more than 75%, 80%, 85%, 90%, 95%, or even more than 99% of their life expectancy. The age of the individual depends on the species of interest. Thus, this percentage is based on the expected life expectancy of the species of interest. For example, in humans, such individuals may be 50 years or older, e.g., 60 years or older, 70 years or older, 80 years or older, 90 years or older, and usually up to 100 years of age, e.g., 90 years of age, i.e., between about 50 and 100 years of age, e.g., 50...55...60...65...70...75...80...85...90...95...100 years or older, or any age between 50 and 1000, suffering from an age-related condition, e.g., cognitive impairment, as described further below, associated with the natural aging process. and individuals of any age who have not yet developed cognitive impairment due to an age-related disease, such as individuals of any age who have not yet developed cognitive impairment, but who have not yet begun to show symptoms of cognitive impairment, such as individuals of about 50 years or older, e.g., 60 years or older, 70 years or older, 80 years or older, 90 years or older, and usually up to 100 years of age, i.e., between about 50 and 100 years of age, e.g., 50...55...60...65...70...75...80...85...90...95...100 years of age, as well as individuals of any age who suffer from cognitive impairment due to an age-related disease, as described below, and individuals of any age who have been diagnosed with an age-related disease generally associated with cognitive impairment, but who have not yet begun to show symptoms of cognitive impairment. Corresponding ages for non-human subjects are known and are intended to apply herein.

[0041] As used herein, "treatment" refers, in appropriate context, to either (i) the prevention of a disease or disorder, or (ii) the reduction or elimination of symptoms of a disease or disorder. Treatment may be effected prophylactically (before the onset of the disease) or therapeutically (after the onset of the disease). The effect may be prophylactic, in the sense of completely or partially preventing the disease or its symptoms, and / or therapeutic, in the sense of partially or completely curing the disease and / or adverse effects that may result from the disease. Thus, the term "treatment" as used herein covers any treatment of age-related diseases or disorders in mammals and includes (a) preventing the onset of the disease in a subject who may be predisposed to the disease but has not yet been diagnosed as having it, (b) arresting the disease, i.e., halting its development, or (c) relieving the disease, i.e., causing regression of the disease. Treatment may result in a variety of different physical manifestations, such as modulation of gene expression, tissue or organ regeneration, etc. Therapeutic agents may be administered before, during, or after the onset of the disease. Of particular interest is the treatment of ongoing disease, where the treatment stabilizes or reduces the patient's undesired clinical symptoms.Such treatment can be performed before the function of the affected tissue is completely lost.The therapy can be administered during the symptomatic stage of the disease, or in some cases, after the symptomatic stage of the disease.

[0042] As used herein, "treatment" refers, in appropriate context, to either (i) the prevention of a disease or disorder, or (ii) the reduction or elimination of symptoms of a disease or disorder. Treatment may be effected prophylactically (before the onset of the disease) or therapeutically (after the onset of the disease). The effect may be prophylactic, in the sense of completely or partially preventing the disease or its symptoms, and / or therapeutic, in the sense of partially or completely curing the disease and / or adverse effects that may result from the disease. Thus, the term "treatment," as used herein, covers any treatment of conditions associated with postoperative recovery in mammals and includes (a) preventing the onset of the condition in a subject, (b) arresting the condition, i.e., halting the condition, or (c) alleviating the condition, i.e., causing regression of the condition. Treatment may result in a variety of different physical manifestations, such as modulation of gene expression, tissue or organ regeneration, reduced inflammation, etc. A therapeutic agent may be administered before, during, or after the onset of the condition. The therapy may be administered during the symptomatic stage of the condition, or optionally after the symptomatic stage of the condition.

[0043] In some embodiments, the age-related condition to be treated is age-related dysfunction in an individual's cognitive abilities. Cognitive abilities, or "cognition," refers to mental processes, including attention and concentration, learning complex tasks and concepts, memory (acquiring, retaining, and recalling new information in the short and / or long term), information processing (handling information gathered by the five senses), visuospatial function (visual perception, depth perception, use of mental imagery, copying drawings, constructing objects or shapes), language production and comprehension, verbal fluency (word finding), problem solving, decision-making, and executive function (planning and prioritization). "Cognitive decline" refers to the progressive decline of one or more of these abilities, such as a decline in memory, language, thinking, judgment, etc. "Cognitive impairment" and "cognitive impairment" refer to a decrease in cognitive ability compared to a healthy individual, e.g., an age-matched healthy individual, or compared to the individual's performance at a previous time, e.g., 2 weeks, 1 month, 2 months, 3 months, 6 months, 1 year, 2 years, 5 years, or 10 years or more ago. "Age-related cognitive impairment" refers to cognitive impairments commonly associated with aging, including, for example, cognitive impairments associated with the natural aging process, e.g., mild cognitive impairment (MCI), and cognitive impairments associated with age-related disorders, i.e., disorders that occur with increasing frequency as the age progresses, e.g., neurodegenerative conditions such as Alzheimer's disease, Parkinson's disease, frontotemporal dementia, Huntington's disease, amyotrophic lateral sclerosis, multiple sclerosis, glaucoma, myotonic dystrophy, and vascular dementia.

[0044] Blood products, including plasma componentsIn practicing the method, a blood product containing a plasma component is administered to an individual in need thereof, e.g., an individual suffering from or at risk of suffering from cognitive impairment and / or age-related dementia or a post-operative condition. Thus, a method according to an embodiment of the present invention involves administering a blood product containing a plasma component from an individual (a "donor individual" or "donor") to an individual (a "recipient individual" or "recipient") who is at least at risk of suffering from or suffering from cognitive impairment and / or age-related dementia or a post-operative condition. A "blood product containing a plasma component" refers to any blood-derived product (e.g., whole blood, plasma, or a fraction thereof) that contains plasma. The term "plasma" is used in its conventional sense to refer to the straw-colored / pale yellow liquid component of blood, which is composed of approximately 92% water, 7% proteins, e.g., albumin, gamma globulin, antihemophilic factor, and other clotting factors, and 1% inorganic salts, sugars, fats, hormones, and vitamins. Non-limiting examples of plasma-containing blood products suitable for use in the present methods include whole blood treated with anticoagulants (e.g., EDTA, citrate, oxalate, heparin, etc.), blood products produced by filtering whole blood to remove white blood cells ("leukoreduction"), blood products consisting of plasmapheretically-derived or apheretically-derived plasma, fresh frozen plasma, blood products consisting essentially of purified plasma, and blood products consisting essentially of plasma fractions. In some cases, the plasma product used is a non-whole blood plasma product, meaning that the product is not whole blood and therefore lacks one or more components found in whole blood, e.g., red blood cells, white blood cells, etc., at least to the extent that these components are present in whole blood. In some cases, the plasma product is substantially, if not completely, acellular, in which case the cell content may be 5% or less by volume, e.g., 1% or less by volume, e.g., 0.5% or less by volume, and in some cases, acellular plasma fractions are compositions that are completely devoid of cells, i.e., they are cell-free.

[0045] Collection of blood products, including plasma components Embodiments of the methods described herein include administering blood products, including plasma components, that may be derived from a donor, such as a human volunteer. The term "human-derived" may refer to such products. Methods for collecting blood products, including plasma, from donors are well known in the art (see, e.g., AABB TECHNICAL MANUAL, (Mark A. Fung, et al., eds., 18th ed. 2014), which is incorporated herein by reference).

[0046] In one embodiment, donations are obtained by venipuncture. In another embodiment, venipuncture is a single venipuncture. In another embodiment, saline replacement is not used. In one embodiment, the process of plasma exchange is used to obtain blood products including plasma. Plasma exchange can involve removing a volume of plasma adjusted to the volume and returning cellular components to the donor. In this embodiment, sodium citrate is used during plasma exchange to prevent cellular coagulation. The volume of plasma collected from the donor is preferably 690-880 mL after citrate administration, and is preferably in line with the donor's body weight.

[0047] 3. Plasma Fraction During World War II, a need arose for a stable plasma expander that could be used when soldiers lost large amounts of blood on the battlefield. As a result, methods for preparing freeze-dried plasma were developed. However, the need for sterile water for reconstitution made the use of freeze-dried plasma difficult in combat situations. Dr. E. J. Cohn suggested that albumin could be used as an alternative and prepared a ready-to-use, stable solution that could be immediately administered for the treatment of shock (see Johan Vandersande, Current Approaches to the Preparation of Plasma Fractions in (Biotechnology of Blood) 165 (Jack Goldstein ed., 1st ed. 1991)). Dr. Cohn's plasma fraction purification procedure utilizes cold ethanol for its denaturing effect and uses changes in pH and temperature to achieve separation.

[0048] One embodiment of the methods described herein involves administering a plasma fraction to a subject. Fractionation is a process in which a specific subset of proteins is separated from plasma. Fractionation techniques are known in the art and are based on steps developed by Cohn et al. in the 1940s (E. Cohn, Preparation and properties of serum and plasma proteins. IV.A system for the separation into fractions of the protein and lipoprotein components of biological tissues and fluids. 68 J Am Chem Soc 459 (1946), incorporated herein by reference). This process involves several steps, each of which uses a specific ethanol concentration, as well as changes in pH, temperature, and osmolality, to selectively precipitate proteins. The precipitate is then separated by centrifugation or sedimentation. The original "Cohn fractionation process" involved separating proteins by precipitation into five fractions designated Fraction I, Fraction II+III, Fraction IV-1, Fraction IV-4, and Fraction V. Albumin was the first identified endpoint (fraction V) product of this process.According to embodiments of the present invention, each fraction (or the effluent from a preceding separation step) contains or may contain a therapeutically useful protein fraction (see Thierry Burnouf, Modern Plasma Fractionation, 21(2) Transfusion Medicine Reviews 101 (2007); Adil Denizli, Plasma fractionation: conventional and chromatographic methods for albumin purification, 4 J. Biol. & Chem. 315, (2011); and T. Brodniewicz-Proba, Human Plasma Fractionation and the Impact of New Technologies on the Use and Quality of Plasma-derived Products, 5 Blood Reviews 245 (1991); and U.S. Pat. Nos. 3,869,431, 5,110,907, 5,219,995, 7,531,513, and 8,772,461, which are incorporated herein by reference). The above experimental parameters may be adjusted to obtain a particular protein fraction.

[0049] In more recent years, fractionation has reached greater complexity and thus constitutes a further embodiment of the present invention. This recent increase in complexity has occurred through the introduction of chromatography techniques resulting in the isolation of new proteins from existing fractions such as cryoprecipitate, cryo-poor plasma, and Cohn fractions; increased IgG recovery through the integration of chromatography and ethanol fractionation processes; and viral reduction / inactivation / removal (ibid.). Anion exchange chromatography can be used to capture proteins at physiological pH and ionic strength, preserving the functional activity of the protein and / or protein fraction. Heparin and monoclonal antibodies are also used in affinity chromatography. Gel filtration fractionation, salt fractionation, and polyethylene glycol fractionation are also used (Hosseini M, Iran J, Biotech, 14(4):213-20(2016)). Those skilled in the art will recognize that the above parameters and techniques can be adjusted to obtain fractions containing specifically desired plasma proteins.

[0050] Plasma fractions can also be ammonium sulfate-based (Odunuga OO, Biochem Compounds, 1:3 (2013); Wingfield PT, Curr Protoc Protein Sci, appx. 3(2001) incorporated herein by reference). In addition to obtaining specific blood fractions, ammonium sulfate-based fractions have been used to deplete abundant proteins from plasma (Saha S, et al., J. Proteomics Bioinform, 5(8)(2012)) incorporated herein by reference).

[0051] In one embodiment of the present invention, plasma is fractionated in an industrial setting. Frozen plasma is thawed at 1°C to 4°C. Continuous refrigerated centrifugation is applied to the thawed plasma to isolate the cryoprecipitate. The collected cryoprecipitate is frozen at or below -30°C and stored. The cryoprecipitate-poor ("cryopoor") plasma is immediately processed (e.g., by primary chromatography) for capture of labile coagulation factors, such as factor IX complex and its components, as well as protease inhibitors, such as antithrombin and C1 esterase inhibitor. Subsequent steps of continuous centrifugation and isolation of the precipitate can be applied. Such techniques are known to those skilled in the art and are described, for example, in U.S. Pat. Nos. 4,624,780, 5,219,995, and 5,288,853, and U.S. Patent Application Nos. 20140343255 and 20150343025, the disclosures of which are incorporated herein by reference in their entireties.

[0052] In one embodiment of the present invention, the plasma fraction may include a plasma fraction containing a significant concentration of albumin. In another embodiment of the present invention, the plasma fraction may include a plasma fraction containing a significant concentration of IgG or intravenous immunoglobulin (IGIV) (e.g., Gamunex-C®). In another embodiment of the present invention, the plasma fraction may include an IGIV plasma fraction, such as Gamunex-C®, that has been substantially depleted of immunoglobulin (IgG) by methods well known to those skilled in the art, such as protein A-mediated depletion (see Keshishian, H., et al., Multiplexed, Quantitative Workflow for Sensitive Biomarker Discovery in Plasma Yields Novel Candidates for Early Myocardial Injury, Molecular & Cellular Proteomics, 14 at 2375-93 (2015)). In a further embodiment, the plasma fraction may be one from which substantially all coagulation factors have been removed, such that the efficacy of the fraction is maintained with a reduced risk of thrombosis. For example, the plasma fraction may be the plasma fraction described in U.S. Patent Application No. 62 / 376,529, filed August 18, 2016, the disclosure of which is incorporated herein by reference in its entirety.

[0053] 4. Albumin products Those skilled in the art will recognize that albumin plasma products ("APP") are broadly classified into two categories: plasma protein fraction (PPF) and human albumin solution (HAS). PPF is derived from a process that results in a higher yield than HAS but has a lower minimum albumin purity than HAS (>83% for PPF and >95% for HAS) (Production of human albumin solution: a continually developing colloid, P. Matejtschuk et al., British Journal of Anaesthesia 85(6):887-95, at 888 (2000)). In some cases, PPF has an albumin purity of 83% to 95% or alternatively 83% to 96%. Albumin purity can be determined by electrophoresis or other quantitative assays, such as mass spectrometry. Furthermore, PPF has been noted by some to be disadvantageous due to the presence of protein "contaminants" such as PKA (ibid.). As a result, PPF preparations have fallen out of favor as albumin plasma products and have even been removed from the pharmacopoeias of certain countries (Id.). Contrary to these concerns, the present invention makes effective use of these "contaminants." In addition to the aforementioned PKA, as well as alpha globulin, beta globulin, and gamma globulin, the methods of the present invention utilize additional proteins or other factors in the "contaminants" that promote processes such as neurogenesis, neuronal survival, and improved cognitive or motor function and reduced neuroinflammation.

[0054] Those skilled in the art will recognize that there are or have been several commercial sources of PPF ("commercial PPF preparations"), including Plasma-Plex™ PPF (Armour Pharmaceutical Co., Tarrytown, NY), Plasmanate™ PPF (Grifols, Clayton, NC), Plasmatein™ (Alpha Therapeutics, Los Angeles, CA), and Protenate™ PPF (Baxter Labs, Inc., Deerfield, IL).

[0055] Those skilled in the art will also recognize that there are or have been several commercial sources of HAS ("commercial HAS preparations"), including Albuminar™ (CSL Behring), AlbuRx™ (CSL Behring), Albutein™ (Grifols, Clayton, NC), Buminate™ (Baxalta, Inc., Bannockburn, IL), Flexbumin™ (Baxalta, Inc., Bannockburn, IL), and Plasbumin™ (Grifols, Clayton, NC).

[0056] A. Plasma Protein Fraction (Human) (PPF) According to the United States Food and Drug Administration ("FDA"), "Plasma Protein Fraction (Human)," or PPF, is the official name for a product defined as "a sterile solution of proteins composed of albumins and globulins derived from human plasma" (Code of Federal Regulations "CFR" 21 CFR 640.90, which is incorporated herein by reference). The source of PPF is plasma recovered from whole blood prepared as specified in 21 CFR 640.1-640.5, which are incorporated herein by reference, or source plasma prepared as specified in 21 CFR 640.60-640.76, which are incorporated herein by reference.

[0057] PPF is tested in accordance with 21 CFR 640.92 (hereby incorporated by reference) to determine that it meets the following criteria: (a) the final product is a 5.0±0.30 percent solution of protein; and (b) The total protein in the final product shall consist of at least 83 percent albumin and not more than 17 percent globulins. Not more than 1 percent of the total protein shall be gamma globulins. Protein composition shall be determined by methods approved for each manufacturer by the Director of the Center for Biologics Evaluation and Research of the Food and Drug Administration.

[0058] As used herein, "plasma protein fraction" or "PPF" refers to a sterile solution of proteins composed of albumin and globulins derived from human plasma, having an albumin content of at least 83%, globulins (including α1 globulin, α2 globulin, β globulin, and γ globulin) and other plasma proteins of not more than 17%, and γ globulins of not more than 1% as determined by electrophoresis (Hink, JH, Jr., et al., Preparation and Properties of a Heat-Treated Human Plasma Protein Fraction, VOX SANGUINIS 2(174)(1957)). PPF may also refer to a solid form having a similar composition when suspended in a solvent. The total globulin fraction can be determined by subtracting albumin from total protein (Busher, J., Serum Albumin and Globulin, CLINICAL METHODS: THE HISTORY, PHYSICAL, AND LABORATORY EXAMINATIONS, Chapter 10, Walker HK, Hall WD, Hurst JD, eds. (1990)).

[0059] B. Albumin (Human) (HAS) According to the FDA, "Albumin (Human)" (also referred to herein as "HAS") is the official name of a product defined as a "sterile solution of albumin derived from human plasma" (Code of Federal Regulations "CFR" 21 CFR 640.80, which is incorporated herein by reference). The source of Albumin (Human) is plasma recovered from whole blood prepared as specified in 21 CFR 640.1-640.5, which are incorporated herein by reference, or source plasma prepared as specified in 21 CFR 640.60-640.76, which are incorporated herein by reference. Other requirements for Albumin (Human) are listed in 21 CFR 640.80-640.84, which are incorporated herein by reference.

[0060] Albumin (human) is tested per 21 CFR 640.82 to determine if it meets the following criteria: (a) Protein Concentration. The final product shall correspond to one of the following concentrations: 4.0±0.25 percent, 5.0±0.30 percent, 20.0±1.2 percent, and 25.0±1.5 percent protein solutions. (b) Protein Composition. At least 96 percent of the total protein in the finished product must be albumin, as determined by a method approved for each manufacturer by the Director of the Center for Biologics Evaluation and Research of the Food and Drug Administration.

[0061] As used herein, "albumin (human)" or "HAS" refers to a sterile solution of proteins composed of albumin and globulins derived from human plasma, with at least 95% albumin content and no more than 5% globulins (including α1 globulin, α2 globulin, β globulin, and γ globulin) and other plasma proteins. HAS may also refer to a solid form with a similar composition when suspended in a solvent. The total globulin fraction may be determined by subtracting albumin from the total protein.

[0062] As will be appreciated by those skilled in the art, the PPF and HAS fractions may be in lyophilized or other solid forms. Such preparations can be used with appropriate additives to form, for example, tablets, powders, granules, or capsules. Solid forms can be formulated into injectable preparations by dissolving, suspending, or emulsifying them in aqueous or non-aqueous solvents, such as vegetable oils or other similar oils, synthetic fatty acid glycerides, esters of higher fatty acids, or propylene glycol, and, if desired, with conventional additives such as solubilizers, isotonicity agents, suspending agents, emulsifiers, stabilizers, and preservatives.

[0063] 5. Fractions with reduced coagulation factors Another embodiment of the present invention uses plasma fractions in which substantially all of the coagulation factors have been removed, such that the efficacy of the fraction is maintained with a reduced risk of thrombosis. Conveniently, blood products may be derived from a young donor or young donor pool and can be devoid of IgM to provide an ABO-compatible young blood product. Currently, transfused plasma is matched to ABO blood types because the presence of naturally occurring antibodies against A and B antigens can result in transfusion reactions. IgM is believed to be a contributing factor to transfusion reactions when ABO-incompatible plasma is given to patients. Removal of IgM from blood products or fractions helps avoid transfusion reactions in subjects receiving the blood products and plasma fractions of the present invention.

[0064] Thus, in one embodiment, the present invention relates to a method of treating or preventing an age-related condition, such as cognitive impairment or neurodegeneration, in a subject. The method comprises administering to the subject a blood product or blood fraction derived from whole blood of an individual or pool of individuals, wherein the blood product or blood fraction substantially lacks (a) at least one clotting factor and / or (b) IgM. In some embodiments, the individual(s) from whom the blood product or blood fraction is derived are young individuals. In some embodiments, the blood product substantially lacks at least one clotting factor and IgM. In certain embodiments, the blood product substantially lacks fibrinogen (Factor I). In further embodiments, the blood product substantially lacks red blood cells and / or white blood cells. In further embodiments, the blood product is substantially acellular. In other embodiments, the blood product is derived from plasma. Such embodiments of the present invention are further supported by U.S. Patent Application No. 62 / 376,529, filed August 18, 2016, the entire contents of which are incorporated herein by reference.

[0065] 6. Protein-enriched plasma protein products Further embodiments of the invention utilize plasma fractions that have reduced albumin concentrations but increased amounts of globulins and other plasma proteins (some termed "contaminants") compared to PPF. These embodiments, like PPF, HAS, Effluent I, and Effluent II / III, are all effectively devoid of coagulation factors. Such plasma fractions are hereafter referred to as "protein-enriched plasma protein products." For example, one embodiment of the invention may utilize a protein-enriched plasma protein product consisting of 82% albumin and 18% α-, β-, and γ-globulins and other plasma proteins. Another embodiment of the invention may utilize a protein-enriched plasma protein product consisting of 81% albumin and 19% α-, β-, and γ-globulins and / or other plasma proteins. Another embodiment of the invention may utilize a protein-enriched plasma protein product consisting of 80% albumin and 20% α-, β-, and γ-globulins and / or other plasma proteins. Further embodiments of the invention may use a protein-enriched plasma protein product consisting of 70-79% albumin and a corresponding 21-30% of α-, β-, and γ-globulins and other plasma proteins. Further embodiments of the invention may use a protein-enriched plasma protein product consisting of 60-69% albumin and a corresponding 31-40% of α-, β-, and γ-globulins and other plasma proteins. Further embodiments of the invention may use a protein-enriched plasma protein product consisting of 50-59% albumin and a corresponding 41-50% of α-, β-, and γ-globulins and other plasma proteins. Further embodiments of the invention may use a protein-enriched plasma protein product consisting of 40-49% albumin and a corresponding 51-60% of α-, β-, and γ-globulins and other plasma proteins. Further embodiments of the invention may use a protein-enriched plasma protein product consisting of 30-39% albumin and a corresponding 61-70% of α-, β-, and γ-globulins and other plasma proteins. A further embodiment of the present invention may use a protein-enriched plasma protein product consisting of 20-29% albumin and a corresponding 71-80% of alpha, beta, and gamma globulins and other plasma proteins.Further embodiments of the invention may use a protein-enriched plasma protein product consisting of 10-19% albumin and a corresponding 81-90% of alpha, beta, and gamma globulins and other plasma proteins. Further embodiments of the invention may use a protein-enriched plasma protein product consisting of 1-9% albumin and a corresponding 91-99% of alpha, beta, and gamma globulins and other plasma proteins. Further embodiments of the invention may use a protein-enriched plasma protein product consisting of 0-1% albumin and 99-100% of alpha, beta, and gamma globulins and other plasma proteins.

[0066] The above-described embodiments of the invention may have a total gamma globulin concentration of 0-5%.

[0067] The specific concentration of a protein in a plasma fraction can be determined using techniques well known to those skilled in the relevant art, including, by way of example and not limitation, electrophoresis, mass spectrometry, ELISA analysis, and Western blot analysis.

[0068] 7. Preparation of Plasma Fractions Methods for preparing PPF and other plasma fractions are well known to those skilled in the art. According to one embodiment of the present invention, blood used to prepare human plasma protein fractions can be collected in flasks containing citrate or anticoagulant citrate dextrose solution (or other anticoagulant) to prevent coagulation, and fractions I, II+III, IV, and PPF can be further separated according to the method disclosed by Hink et al. (See Hink, JH, Jr., et al., Preparation and Properties of a Heat-Treated Human Plasma Protein Fraction, VOX SANGUINIS 2(174) (1957), incorporated herein by reference). According to this method, the mixture can be collected at 2-8°C. The plasma can then be separated by centrifugation at 7°C, removed, and stored at -20°C. The plasma can then be thawed at 37°C and fractionated, preferably within 8 hours of removal from -20°C storage.

[0069] Plasma can be separated from Fraction I using 8% ethanol at a protein concentration of 5.1 to 5.6 percent, pH 7.2, and a temperature of -2 to -2.5°C. Cold 53.3% ethanol (176 mL / L of plasma) can be added along with acetate buffer (200 mL of 4 M sodium acetate, 230 mL of glacial acetic acid, and H2O up to 1 L) using a jet at a rate of, for example, 450 mL / min, while the plasma temperature is lowered to -2°C. Fraction I can be separated and isolated from the effluent (Effluent I) by ultracentrifugation. Fibrinogen can be obtained from Fraction I according to methods well known to those skilled in the art.

[0070] Fraction II+III can be separated from Effluent I by adjusting the effluent to 21 percent ethanol at a protein concentration of 4.3 percent and a temperature of -6°C at pH 6.8. While lowering the temperature of Effluent I to -6°C, 95 percent cold ethanol (176 mL / L of Effluent I) can be added, for example, using a jet at a rate of 500 mL / min, along with 10 M acetic acid used for pH adjustment. The resulting precipitate (Fraction II+III) can be removed by centrifugation at -6°C. Gamma globulin can be obtained from Fraction II+III using methods well known to those skilled in the art.

[0071] Fraction IV-1 can be isolated from Effluent II+III ("Effluent II / III") by adjusting the effluent to 19% ethanol at a protein concentration of 3% and a pH of 5.2 at -6°C. While Effluent II / III is maintained at -6°C for 6 hours, HO and 10 M acetic acid, used for pH adjustment, can be added using a jet. The precipitated Fraction VI-1 can be allowed to settle at -6°C for 6 hours and then separated from the effluent by centrifugation at the same temperature. At a protein concentration of 2.5% and a pH of 4.65, a temperature of -7°C, and a stable plasma protein fraction can be recovered from Effluent IV-1 by adjusting the ethanol concentration to 30%. This can be achieved by adjusting the pH of Effluent IV-1 with cold acid-alcohol (2 parts 2 M acetic acid and 1 part 95% ethanol). While maintaining the temperature at -7°C, 170 mL of cold ethanol (95%) is added for every liter of adjusted Effluent IV-1. Precipitating proteins can be removed by centrifugation at -7°C after settling for 36 hours.

[0072] The recovered protein (stable plasma protein fraction) can be dried (e.g., by lyophilization) to remove alcohol and HO. The resulting dried powder can be dissolved in sterile distilled water, e.g., using 15 liters of water per kg of powder, and the solution adjusted to pH 7.0 with 1 M NaOH. A final protein concentration of 5 percent can be achieved by adding sterile distilled water containing sodium acetyltryptophan, sodium caprylate, and NaCl to a final concentration of 0.004 M acetyltryptophan salt, 0.004 M caprylate, and 0.112 M sodium. Finally, the solution can be filtered at 10°C to obtain a clear solution, followed by heat treatment at 60°C for at least 10 hours to inactivate pathogens.

[0073] Those skilled in the art will recognize that each of the various fractions and effluents described above can be used in conjunction with the methods of the present invention to treat conditions associated with disease or post-surgical recovery. For example, and not by way of limitation, Effluent I or Effluent II / III can be utilized to treat conditions associated with disease or post-surgical recovery, such as cognitive impairment and neurodegenerative disorders, or to promote post-surgical recovery, and are embodiments of the present invention.

[0074] The above-described methods for preparing plasma fractions and plasma protein fractions (PPF) are merely exemplary and relate to embodiments of the present invention. Those skilled in the art will recognize that these methods can be modified. For example, in various embodiments and methods of the present invention, various variations of plasma fractions and plasma protein fractions can be produced by adjusting, among other things, pH, temperature, and ethanol concentration. In another example, a further embodiment of the present invention contemplates the use of nanofiltration for the removal / inactivation of pathogens in plasma fractions and plasma protein fractions.

[0075] Further embodiments of the present invention contemplate methods and compositions that use and / or include additional plasma fractions. For example, the present invention demonstrates that a particular concentration of albumin is not required to, inter alia, improve cognitive activity or treat conditions associated with post-operative recovery or promote post-operative recovery. Thus, fractions with reduced albumin concentrations, e.g., fractions with 83% or less albumin, are contemplated by the present invention.

[0076] 8. Treatment Aspects of the methods of the invention described herein include treating a subject with a plasma-containing blood product, e.g., a plasma fraction, e.g., as described above. One embodiment includes treating a human subject with a plasma-containing blood product. Those skilled in the art will recognize that methods of treating a subject with plasma-containing blood products are known in the art. By way of example and not limitation, one embodiment of the methods of the invention described herein includes administering fresh frozen plasma to a subject for the treatment and / or prevention of conditions associated with cognitive impairment and / or age-related dementia or post-surgical recovery. In one embodiment, the plasma-containing blood product is administered immediately, e.g., within about 12-48 hours after collection from the donor, to an individual suffering from or at risk for a condition associated with cognitive impairment and / or age-related dementia or post-surgical recovery. In another embodiment, the product can be stored under refrigeration, e.g., at 0-10°C. In another embodiment, the fresh frozen plasma is stored frozen (cryopreserved) at -18°C or below. The fresh frozen plasma is thawed prior to administration and administered to the subject upon thawing, 60-75 minutes after the thawing process begins. Each subject preferably receives a single unit of fresh frozen plasma (200-250 mL). This fresh frozen plasma is preferably derived from donors within a predetermined age range. In one embodiment of the present invention, the fresh frozen plasma is donated (derived) from a young individual. In another embodiment of the present invention, the fresh frozen plasma is donated (derived) from a donor of the same gender. In another embodiment of the present invention, the fresh frozen plasma is donated (derived) from a donor within the age range of 18-22 years.

[0077] In an embodiment of the invention, the compositions of the invention (e.g., plasma-containing blood products, such as plasma fractions) are administered intravenously. The compositions of the invention may also be delivered intraperitoneally. In other embodiments of the invention, the compositions of the invention may be delivered orally, subcutaneously, or topically. Topical preparations for treating and enhancing wounds are known as gels, creams, ointments, gauzes, patches, and the like, and the compositions of the invention may be formulated as such (see, e.g., Kahn Aw, et al., Pharmacogn Mag, 9 (Suppl 1):S6-S10 (2013); U.S. Pat. No. 5,641,483; U.S. Pat. No. 4,885,163; U.S. Pat. No. 8,313,764).

[0078] In one embodiment of the present invention, blood products, including plasma, are screened by blood type after donation. In another embodiment of the present invention, blood products, including plasma, are screened for infectious agents, such as HIV I&II, HBV, HCV, HTLV I&II, and anti-HBc, in accordance with the requirements of 21 CFR 640.33 and recommendations contained in FDA guidance documents.

[0079] In yet another embodiment of the invention, the subject is treated with a "plasma fraction." In one embodiment of the invention, the plasma fraction is PPF or HAS. In a further embodiment of the invention, the plasma fraction is one of a commercial PPF preparation or a commercial HAS preparation. In another embodiment of the invention, the plasma fraction is PPF or HAS derived from a pool of individuals in a particular age range, such as young individuals, or is a modified PPF or HAS fraction (e.g., PPF or HAS from which one or more specific proteins have been partially or substantially removed) that has been subjected to further fractionation or processing. In another embodiment of the invention, the plasma fraction is an IGIV plasma fraction that is substantially depleted of immunoglobulins (IgG). A blood fraction that is "substantially depleted" or "substantially removed" of a particular protein, such as IgG, refers to a blood fraction that contains less than about 50%, e.g., less than 45%, 40%, 35%, 30%, 25%, 20%, 15%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.25%, 0.1%, an undetectable level, or any integer between these values, of the amount occurring in a reference product or whole blood plasma, as measured using standard assays well known in the art.

[0080] Aspects of the methods of the invention described herein include treating a subject with a plasma-containing blood product, e.g., plasma or a plasma fraction, e.g., as described above. One embodiment includes treating a human subject with a plasma-containing blood product. Those skilled in the art will recognize that methods of treating a subject with plasma-containing blood products are recognized in the art. By way of example and not limitation, one embodiment of the methods of the invention described herein includes administering fresh frozen plasma to a subject to treat a condition associated with post-surgical recovery. In one embodiment, the plasma-containing blood product is administered immediately, e.g., within about 12 to 48 hours after collection from the donor, to an individual suffering from an undesirable condition associated with post-surgical recovery.

[0081] In another embodiment, the product may be stored under refrigeration, for example, at 0-10°C. In another embodiment, the fresh frozen plasma is stored frozen (cryopreserved) at -18°C or below. The fresh frozen plasma is thawed prior to administration and administered to the subject immediately, 60-75 minutes after the thawing process begins. Each subject preferably receives a single unit of fresh frozen plasma (200-250 mL). This fresh frozen plasma is preferably derived from donors within a predetermined age range. In one embodiment of the invention, the fresh frozen plasma is donated (derived from) a young individual. In another embodiment of the invention, the fresh frozen plasma is donated (derived from) a donor of the same gender. In another embodiment of the invention, the fresh frozen plasma is donated (derived from) a donor within the age range of 18-22 years. In one embodiment, the subject is treated twice weekly with 3-4 days between infusions. In one embodiment of the invention, treatment is continued until a specific endpoint is reached.

[0082] In one embodiment of the present invention, blood products, including plasma, are screened by blood type after donation. In another embodiment of the present invention, blood products, including plasma, are screened for infectious agents, such as HIV I&II, HBV, HCV, HTLV I&II, and anti-HBc, in accordance with the requirements of 21 CFR 640.33 and recommendations contained in FDA guidance documents.

[0083] In yet another embodiment of the present invention, the subject is treated with a "plasma fraction." In one embodiment of the present invention, the plasma fraction is PPF or HAS. In a further embodiment of the present invention, the plasma fraction is one of a commercial PPF preparation or a commercial HAS preparation. In another embodiment of the present invention, the plasma fraction is PPF or HAS derived from a pool of individuals of a particular age range, such as young individuals, or is a modified PPF or HAS fraction (e.g., PPF or HAS from which one or more specific proteins have been partially or substantially removed) that has been subjected to further fractionation or processing. In another embodiment of the present invention, the plasma fraction is an IGIV plasma fraction that is substantially depleted of immunoglobulins (IgG). A blood fraction that is "substantially depleted" or "substantially removed" of a particular protein, such as IgG, refers to a blood fraction that contains less than about 50%, e.g., less than 45%, 40%, 35%, 30%, 25%, 20%, 15%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.25%, 0.1%, an undetectable level, or any integer between these values, of the amount occurring in a reference product or whole blood plasma, as measured using standard assays well known in the art.

[0084] 9. Monitoring Another aspect of the present invention relates to a method for monitoring the effects of a drug on a subject for the treatment of cognitive impairment and / or age-related dementia, comprising comparing cognitive function before and after treatment. Those skilled in the art will recognize that there are well-known methods for assessing cognitive function. For example, but not by way of limitation, the method may include assessment of cognitive function based on a medical history, family history, physical and neurological examinations by a clinician specializing in dementia and cognitive function, laboratory tests, and neuropsychological evaluation. Further embodiments contemplated by the present invention include assessment of state of consciousness using, for example, the Glasgow Coma Scale (EMV), mental status tests including the Abbreviated Mental Test Score (AMTS) or the Mini-Mental State Examination (MMSE) (Folstein et al., J. Psychiatr. Res 1975;12:1289-198), a global assessment of higher-order functions, and estimation of intracranial pressure by funduscopy, etc.

[0085] In one embodiment, peripheral nervous system testing can be used to assess cognitive function, including smell, visual field and vision, eye movements and pupils (sympathetic and parasympathetic), facial sensory function, strength of the muscles of the face and shoulder girdle, hearing, taste, pharyngeal movements and reflexes, tongue movements (which can be tested individually (e.g., visual acuity can be tested with a Snellen eye chart, and reflexes including the masseter, biceps, and triceps tendons, hamstrings, Achilles tendon reflex, and plantar reflex (i.e., Babinski sign) are tested with the use of a reflex hammer)), muscle strength, muscle tone, and signs of rigidity, frequently on an MRC scale of 1-5.

[0086] 10. Administration In practicing the methods of the present invention, a plasma fraction is administered to a subject. In one embodiment, the plasma fraction is administered by intravenous infusion. The rate of infusion can vary, but in one embodiment of the present invention, the infusion rate is 5-8 mL / min. Those skilled in the art will recognize that the infusion rate can depend on the subject's condition and response to administration.

[0087] In embodiments in which an effective amount of an active agent is administered to an adult mammal, the amount or dosage is effective if it is administered for a suitable period, such as one week or more, for example, two weeks or more, for example, three weeks or more, one month or more, two months or more, three months or more, four months or more, five months or more, six months or more, one year or more, etc., and results in a demonstrated reduction in the condition of the adult mammal, such as cognitive impairment, or a delay in cognitive impairment and / or cognitive improvement. For example, an effective dose is one that, when administered over a suitable period, slows cognitive decline by, for example, about 20% or more, e.g., 30% or more, 40% or more, or 50% or more, and in some cases, 60% or more, 70% or more, 80% or more, or 90% or more. For example, administering an effective amount of plasma to a patient suffering from a condition associated with natural aging or age-related disorders or post-operative recovery can halt cognitive decline. In some cases, an effective amount or dose of a blood product not only slows or halts the progression of the condition, but also induces a reversal of the condition, i.e., results in improved cognitive performance. For example, in some cases, an effective amount is an amount that, when administered for a suitable period, usually at least about 1 week, possibly about 2 weeks or more, depending on the individual, about 3 weeks, 4 weeks, 8 weeks or more, improves the cognitive performance of an individual suffering from age-related cognitive impairment, for example, 1.5 times, 2 times, 3 times, 4 times, 5 times, in some cases 6 times, 7 times, 8 times, 9 times, or 10 times or more, compared to the cognition before administration of blood products or fractions.In some cases, an effective amount or effective dose of an active agent not only slows or stops the progression of the disease state, but also induces the reversal of the condition, i.e., leads to the improvement of cognitive function.For example, in some cases, an effective amount is an amount that, when administered for a suitable period, improves the symptoms of an individual suffering from cognitive decline or impairment, for example, 1.5 times, 2 times, 3 times, 4 times, 5 times, in some cases 6 times, 7 times, 8 times, 9 times, or 10 times or more, compared to the untreated individual before administration of the drug.

[0088] In other embodiments, the blood plasmin fraction or plasma fraction is administered according to one or more dosing regimens described in U.S. Patent Application No. 62 / 490,519, the entire disclosure of which is incorporated herein by reference. Accordingly, one embodiment of the present invention comprises treating a subject diagnosed with a condition associated with cognitive impairment or post-surgical recovery by administering to the subject an effective amount of plasma or plasma fraction, wherein the plasma or plasma fraction is administered to result in improved cognitive function or neurogenesis or improved wound healing, presence of markers, reduced pain, or reduced inflammation after the mean or median half-life of the plasma protein or plasma fraction protein is reached, compared to a most recent dose (referred to herein as "pulse dosing" or "pulse-dosed"). (See U.S. Patent No. 10,357,513 and U.S. Patent Application Nos. 15 / 961,618 and 62 / 701,411, the disclosures of which are incorporated herein by reference in their entireties.)

[0089] Other embodiments of the invention include administering an effective amount of plasma or a plasma fraction and then monitoring the subject for improved function, wound healing, the presence of markers, reduced pain, or reduced inflammation.

[0090] Other embodiments of the invention include administering plasma or a plasma fraction according to a dosing regimen of at least two consecutive days and monitoring the subject for improved cognitive function or HSC marker levels at least three days after the last administration. Further embodiments of the invention include administering plasma or a plasma fraction according to a dosing regimen of at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 consecutive days and monitoring the subject for cognitive function, wound healing, presence of markers, reduced pain, or reduced inflammation at least three days after the last administration. Yet another embodiment of the invention includes administering plasma or a plasma fraction according to a dosing regimen of at least two consecutive days and monitoring for cognitive improvement, improved function, wound healing, presence of markers, reduced pain, or reduced inflammation after the last administration date, for the average half-life of proteins in the plasma or plasma fraction. Another embodiment of the present invention involves administering plasma or a plasma fraction according to a dosing regimen of 2 to 14 consecutive days, where each interval between doses may be 0 to 3 days.

[0091] In some cases, pulse dosing according to the present invention involves administration of a first dose set, e.g., as described above, followed by a dosing-free period, e.g., a "drug-free period," followed by administration of another dose or dose set. The length of this "drug-free" period can vary, but in some embodiments is 7 days or more, e.g., 10 days or more, e.g., 14 days or more, and in some cases the drug-free period extends from 15 to 365 days, e.g., 30 to 90 days, e.g., 30 to 60 days. Thus, embodiments of the present methods involve non-chronic (i.e., non-sustained) dosing, e.g., non-chronic administration of a plasma product. In some embodiments, the pattern of pulse dosing followed by a drug-free period is repeated as many times as desired, and in some cases, the pattern continues for one year or more, e.g., two years or more, up to and including the lifetime of the subject. Another embodiment of the invention involves administering plasma or a plasma fraction according to a 5-day consecutive dosing regimen, which includes a 2-3 day drug-free period followed by 2-14 consecutive days of dosing.

[0092] Biochemically, an "effective amount" or "effective dose" of an active agent refers to an amount of active agent that prevents, antagonizes, reduces, decreases, or inhibits cognitive impairment or age-related dementia by about 20% or more, e.g., 30% or more, 40% or more, or 50% or more, and in some cases 60% or more, 70% or more, 80% or more, or 90% or more, and in some cases about 100%, i.e., to a negligible extent, and in some cases reverses its progression or reverses undesirable conditions associated with post-operative recovery.

[0093] 11. Plasma Protein Fractions In practicing the methods of the present invention, a plasma fraction is administered to a subject. In one embodiment, the plasma fraction is a plasma protein fraction (PPF). In a further embodiment, the PPF is selected from a commercial PPF preparation.

[0094] In another embodiment, PPF contains 88% normal human albumin, 12% alpha and beta globulins, and 1% or less gamma globulins, as determined by electrophoresis. This embodiment, used in the practice of the methods of the invention, includes, for example, the use of PPF as a 5% solution buffered with sodium carbonate and stabilized with 0.004 M sodium caprylate and 0.004 M acetyltryptophan. Additional formulations, such as those that modify the concentration of solvents and stabilizers as well as the percentage of PPF in solution (e.g., from about 1% to about 10%, from about 10% to about 20%, from about 20% to 25%, from about 25% to 30%), may also be utilized in the practice of the methods of the invention.

[0095] 12. Plasma fractions of specific donor ages One embodiment of the present invention involves administering a plasma fraction or plasma fraction derived from the plasma of individuals within a particular age range. A further embodiment of the present invention involves administering a plasma protein fraction derived from the plasma of individuals within a particular age range. One embodiment involves administering PPF or HAS derived from the plasma of young individuals. In another embodiment of the present invention, the young individuals are of a single particular age or a particular age range. In yet another embodiment, the average age of the donors is less than the age of the subject or less than the average age of the subjects being treated.

[0096] Certain embodiments of the present invention involve pooling blood or plasma from individuals of a particular age range and fractionating the plasma as described above to obtain a plasma protein fraction product, such as PPF or HAS. In alternative embodiments of the present invention, the plasma protein fraction or a particular plasma protein fraction is obtained from a particular individual falling within a specified age range. In another embodiment of the present invention, the plasma fraction, plasma fraction, or a particular plasma protein fraction product is obtained from a pool of younger individuals, where "young" may be determined by chronological age or biological age as described above, and the age(s) of the individuals may be of a particular age or age range.

[0097] 13. Indications The methods and blood products and fractions, including plasma, are useful for treating, e.g., preventing, age-related conditions, such as impairment of an individual's cognitive abilities, e.g., cognitive impairment, including, but not limited to, age-related dementia, immunological conditions, cancer, and physical and functional decline. Individuals suffering from or at risk of developing age-related cognitive impairment and who may benefit from treatment with the blood products, including plasma, of the present invention, e.g., according to the methods disclosed herein, include individuals aged about 50 years or older, e.g., 60 years or older, 70 years or older, 80 years or older, 90 years or older, and 100 years or older, i.e., between about 50 and 100 years old, e.g., 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or about 100 years old, Cognitive impairments associated with the natural aging process include individuals suffering from mild cognitive impairment (MCI), and individuals about 50 years of age or older, e.g., 60 years of age or older, 70 years of age or older, 80 years of age or older, 90 years of age or older, and typically under 100 years of age, i.e., about 50-90 years of age, e.g., 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or about 100 years of age, who have not yet begun to show symptoms of cognitive impairment. Examples of cognitive impairments due to natural aging include:

[0098] A. Mild cognitive impairment Mild cognitive impairment (MCI) is a moderate disruption of cognition that manifests as problems with memory or other mental functions, such as planning, following instructions, or decision-making, over time, while overall mental function and everyday activities remain intact. Thus, although significant neuronal death does not generally occur, neurons in the aging brain are susceptible to sublethal age-related changes in structure, synaptic integrity, and molecular processing at synapses, all of which impair cognitive function.

[0099] Individuals suffering from or at risk of developing age-related cognitive impairment and who may benefit from treatment with the blood products or fractions, including plasma, of the present invention, e.g., according to the methods disclosed herein, include individuals of any age who suffer from cognitive impairment resulting from an age-related disorder, as well as individuals of any age who have been diagnosed with an age-related disorder commonly associated with cognitive impairment, but who have not yet begun to exhibit symptoms of cognitive impairment. Examples of such age-related disorders include:

[0100] B. Alzheimer's disease Alzheimer's disease is a progressive and unstoppable loss of cognitive function associated with excessive numbers of senile plaques in the cerebral cortex and subcortical gray matter, as well as neurofibrillary tangles composed of β-amyloid and tau proteins. Common forms affect people over the age of 60, and its incidence increases with age. Alzheimer's disease accounts for more than 65% of dementia in older adults.

[0101] The cause of Alzheimer's disease is unknown. The disease is familial in approximately 15-20% of cases. The remaining so-called sporadic cases have some genetic determinant. The disease has an autosomal dominant genetic pattern in most early-onset cases and some late-onset cases, with variable penetrance in later life. Environmental factors are the focus of active investigation.

[0102] During the course of the disease, synaptic and ultimately neuronal loss occurs in the cerebral cortex, hippocampus, and subcortical structures (including selective cell loss in the nucleus basalis of Meynert), the locus coeruleus, and the dorsal raphe nucleus. Cerebral glucose utilization and perfusion are reduced in some brain regions (parietal and temporal cortices in early disease, prefrontal cortex in late disease). The pathogenesis of Alzheimer's disease involves the formation of neuritic or senile plaques (composed of neurites, astrocytes, and glial cells around an amyloid core) and neurofibrillary tangles (composed of paired helical filaments). Senile plaques and neurofibrillary tangles occur with normal aging but are much more common in people with Alzheimer's disease.

[0103] C. Parkinson's disease Parkinson's disease (PD) is an idiopathic, slowly progressive, degenerative CNS disorder characterized by slowed movements, muscle rigidity, resting tremor, and postural instability. Originally considered primarily a motor disorder, PD is now recognized to also affect cognition, behavior, sleep, autonomic function, and sensory function. The most common cognitive impairments include impairments in attention and concentration, working memory, executive function, language production, and visuospatial function.

[0104] In primary Parkinson's disease, pigmented neurons in the substantia nigra, locus coeruleus, and other brainstem dopaminergic cell groups are lost. The cause is unknown. Loss of nigral neurons that project to the caudate nucleus and putamen leads to depletion of the neurotransmitter dopamine in these areas. Onset is generally after age 40, with an increased incidence in older populations.

[0105] Secondary parkinsonism results from the loss or disruption of dopamine action in the basal ganglia due to other idiopathic degenerative diseases, drugs, or exogenous toxins. The most common cause of secondary parkinsonism is the ingestion of antipsychotic drugs or reserpine, which block dopamine receptors, resulting in parkinsonism. Less common causes include carbon monoxide or manganese poisoning, hydrocephalus, structural lesions (tumors, infarctions affecting the midbrain or basal ganglia), subdural hematomas, and degenerative disorders including striatonigral degeneration.

[0106] D. frontotemporal dementia Frontotemporal dementia (FTD) is a condition that results from the progressive deterioration of the frontal lobes of the brain. Over time, degeneration may progress to the temporal lobes. Second only to Alzheimer's disease (AD) in prevalence, FTD accounts for 20% of presenile dementia cases. Symptoms are divided into three groups based on the function of the frontal and temporal lobes.

[0107] Symptoms of behavioral FTD (bvFTD) include lethargy and loss of spontaneity on the one hand, and disinhibition on the other. In progressive non-fluent aphasia (PNFA), speech fluency is disrupted due to articulation difficulties and phonological and / or grammatical errors, but comprehension of language is preserved. In semantic dementia (SD), patients maintain fluency with normal phonology and grammar, but naming and comprehension of language become increasingly difficult. Other cognitive symptoms common to all FTD patients include impairments in executive function and concentration. Other cognitive abilities, including perception, spatial ability, memory, and executive functioning, generally remain intact. FTD can be diagnosed by observing atrophy of the frontal and / or anterior temporal lobes, as revealed by structural MRI scans.

[0108] FTD exists in several forms, all of which can be treated or prevented using the present methods and compositions. For example, one form of frontotemporal dementia is semantic dementia (SD). SD is characterized by loss of semantic memory in both verbal and nonverbal domains. Patients with SD often complain of word-finding difficulties. Clinical signs include fluent aphasia, anomia, impaired comprehension of word meanings, and associative visual agnosia (an inability to match semantically related pictures or objects). While cases of "pure" semantic dementia with few subsequent behavioral symptoms have been described, progression of the disease often results in behavioral and personality changes similar to those seen in frontotemporal dementia. Structural MRI imaging shows a characteristic pattern of temporal lobe atrophy (predominantly left), with inferior lesions greater than superior lesions and greater anterior temporal lobe atrophy than posterior temporal lobe atrophy.

[0109] As another example, another form of frontotemporal dementia is Pick's disease (PiD, also known as PcD). The defining feature of this disease is the accumulation of tau protein in neurons, forming silver-staining spherical aggregates known as "Pick bodies." Symptoms include loss of speech (aphasia) and dementia. Patients with orbitofrontal dysfunction can become aggressive and socially inept. They may steal or exhibit compulsive or repetitive stereotyped behaviors. Patients with dorsomedial or dorsolateral frontal dysfunction may exhibit apathy, blunted affect, or reduced spontaneity. Patients may exhibit a lack of self-monitoring, abnormal self-awareness, and an inability to comprehend meaning. Patients with gray matter loss in the bilateral posterolateral orbitofrontal cortex and right anterior insula may exhibit altered eating behavior, such as a pathological sweet preference. Patients with more focal gray matter loss in the anterior lateral orbitofrontal cortex may develop binge eating. Although some symptoms may initially be alleviated, the disease progresses and patients often die within 2 to 10 years.

[0110] E. Huntington's disease Huntington's disease (HD) is a hereditary, progressive neurodegenerative disorder characterized by the development of emotional, behavioral, and psychiatric disorders, loss of intellectual or cognitive function, and movement abnormalities (dyspraxia). Typical signs of HD include the onset of chorea (involuntary, rapid, irregular, jerky movements that may affect the face, arms, legs, or trunk) and cognitive decline, including a gradual loss of thought processing and acquired intellectual skills. Impairments in memory, abstract thinking, and judgment, a loss of sense of time, place, or identity (disorientation), increased agitation, and personality changes (disorganization) may occur. Symptoms typically become apparent during the fourth or fifth decade, but the age at onset can vary, ranging from early childhood to late adulthood (e.g., the seventies or eighties).

[0111] HD is transmitted in families as an autosomal dominant trait. The disorder results from an abnormally long sequence, or "repeat," of coded instructions in a gene on chromosome 4 (4p16.3). The progressive loss of nervous system function associated with HD results from the loss of neurons in certain regions of the brain, including the basal ganglia and cerebral cortex.

[0112] F. Amyotrophic lateral sclerosis Amyotrophic lateral sclerosis (ALS) is a rapidly progressive and always fatal neurological disease that affects motor neurons. Muscle weakness and atrophy and signs of anterior horn cell dysfunction are most often initially seen in the hands and less commonly in the feet. The site of onset is random, and progression is asymmetric. Muscle cramps are common and may precede weakness. Patients rarely survive for 30 years; 50% die within 3 years of onset, 20% survive for 5 years, and 10% survive for 10 years.

[0113] Diagnostic features include onset in mid- or late adulthood and progressive generalized movement disorders without sensory abnormalities. Nerve conduction velocities remain normal until late in the disease. Recent studies have also documented symptoms of cognitive dysfunction, particularly declines in immediate verbal memory, visual memory, language, and executive function.

[0114] In ALS patients, reductions in cell body area, synapse number, and total synaptic length have been reported, even in neurons that appear normal. It has been suggested that functional impairment may occur due to continued synapse loss once active zone plasticity reaches its limit. Promotion of new synapse formation or prevention of synapse loss may preserve neuronal function in these patients.

[0115] G. Multiple sclerosisMultiple sclerosis (MS) is characterized by a variety of symptoms and signs of CNS dysfunction, with recurring remissions and exacerbations. The most common presenting symptoms are paresthesias in one or more limbs, the trunk, or one side of the face; weakness or ataxia in the legs or hands; or visual disturbances, such as partial blindness and pain in one eye (retrobulbar optic neuritis), dim vision, or a scotoma. Common cognitive dysfunctions include impairments in memory (acquiring, retaining, and recalling new information), attention and concentration (especially divided attention), information processing, executive function, visuospatial function, and verbal fluency. Common early symptoms include ophthalmoplegia resulting in double vision (diplopia), transient weakness in one or more limbs, slight stiffness or unusual fatigability in the hands and feet, minor gait disturbances, difficulty with bladder control, dizziness, and mild emotional disturbances, all of which indicate diffuse CNS damage and often occur months or years before the disease is recognized. Excessive fever can accentuate symptoms and signs.

[0116] The course is highly variable and unpredictable and is remittent in most patients. Initially, there may be months or years of remission between episodes, especially if the disease begins with retrobulbar optic neuritis. However, some patients have frequent attacks and become rapidly incapacitating, and in a minority the course may be rapidly progressive.

[0117] H. glaucoma Glaucoma is a common neurodegenerative disease affecting retinal ganglion cells (RGCs). Evidence supports the existence of a compartmentalized degenerative program in synapses and dendrites containing RGCs. Recent evidence has also demonstrated a correlation between cognitive impairment and glaucoma in older adults (Yochim BP, et al. Prevalence of cognitive impairment, depression, and anxiety symptoms among older adults with glaucoma. J Glaucoma. 2012;21(4):250-254).

[0118] I. Myotonic dystrophyMyotonic dystrophy (DM) is an autosomal dominant, multisystem disorder characterized by dystrophic muscle weakness and myotonia. The molecular defect is a trinucleotide (CTG) repeat expansion in the 3' untranslated region of the myotonin protein kinase gene on chromosome 19q. Symptoms can occur at any age, and there is a wide range of clinical severity. Myotonia is prominent in the hand muscles, and ptosis is common even in mild cases. Severe cases develop significant peripheral muscle weakness and are often accompanied by cataracts, premature hair loss, axe-like facies, cardiac arrhythmias, testicular atrophy, and endocrine abnormalities (e.g., diabetes mellitus). Mental retardation is common in severe congenital forms, while milder adult forms of the disorder exhibit age-related decline in frontal and temporal cognitive function, particularly language and executive function. Severely affected individuals die by their early 50s.

[0119] J. Dementia Dementia refers to a class of disorders whose symptoms affect thinking and social skills severely enough to interfere with daily functioning. Other cases of dementia include vascular dementia, discussed below, and dementia with Lewy bodies, in addition to the dementia observed in the later stages of age-related disorders discussed above.

[0120] In vascular dementia or "multi-infarct dementia," cognitive impairment is caused by problems with the brain's blood supply, typically through a series of minor strokes, or sometimes by one major stroke that precedes or follows other, less severe strokes. Vascular lesions can be the result of cerebrovascular disease, such as small vessel disease, or focal lesions, or both. Patients with vascular dementia present with acute or subacute cognitive impairment after an acute cerebrovascular event, followed by progressive cognitive decline. Cognitive impairment is similar to that observed in Alzheimer's disease, including impairments in language, memory, complex visual processing, or executive function, but the associated brain changes are due to chronically reduced cerebral blood flow, which ultimately leads to dementia, rather than AD pathology. Neuroimaging with single-photon emission computed tomography (SPECT) and positron emission tomography (PET) can be used in conjunction with mental status evaluation to confirm a diagnosis of multi-infarct dementia.

[0121] Dementia with Lewy bodies (DLB, also known by various other names, including dementia with Lewy bodies, diffuse Lewy body disease, cortical Lewy body disease, and senile dementia of the Lewy type) is a type of dementia characterized anatomically by the presence of Lewy bodies (clusters of alpha-synuclein and ubiquitin proteins) in neurons, detectable on histological examination of postmortem brain. Its main feature is cognitive decline, particularly of executive function. Attention and short-term memory are variable.

[0122] Persistent or recurrent visual hallucinations with vivid and detailed descriptions are often early diagnostic symptoms. DLB is often confused with Alzheimer's disease and / or vascular dementia in its early stages, but whereas Alzheimer's disease has a fairly gradual onset, DLB often has a rapid or acute onset. DLB symptoms also include motor symptoms similar to those of Parkinson's disease. DLB is distinguished from the dementia that sometimes occurs in Parkinson's disease by the time frame in which dementia symptoms appear relative to Parkinson's disease symptoms. Parkinson's disease with dementia (POD) is diagnosed when the onset of dementia occurs more than one year after the onset of Parkinson's disease. DLB is diagnosed when cognitive symptoms begin simultaneously with or within one year of Parkinson's disease symptoms.

[0123] K. progressive supranuclear palsy Progressive supranuclear palsy (PSP) is a brain disorder that causes severe, progressive problems with gait and balance control, along with complex eye movements and thinking problems. One of the disease's hallmark symptoms is an inability to properly aim the eyes, due to lesions in the area of ​​the brain that coordinates eye movements. Some individuals describe this effect as blurred vision. Affected individuals often exhibit mood and behavior changes, including depression and apathy, as well as progressive mild dementia. The disorder's long name indicates that the disease begins slowly, continues to worsen (progressive), and causes weakness (paralysis) by damaging specific areas of the brain (supranuclear regions) above the pea-sized structures called nuclei that control eye movement. PSP was first described as a distinct disorder in 1964, when three scientists published a paper distinguishing the condition from Parkinson's disease. PSP is sometimes called Steele-Richardson-Olszewski syndrome, reflecting a combination of the names of the scientists who defined the disorder. PSP gradually worsens, but no one dies from it.

[0124] L. ataxiaPeople with ataxia have problems with coordination because the parts of the nervous system that control movement and balance are affected. Ataxia can affect fingers, hands, arms, legs, body, speech, and eye movements. The term ataxia is often used to describe symptoms of inability to coordinate that may be related to infection, injury, other disease, or degenerative changes in the central nervous system. Ataxia is also used to describe a group of specific degenerative disorders of the nervous system called hereditary and sporadic ataxias, which are the primary focus of the National Ataxia Foundation.

[0125] M. multiple system atrophy Multiple system atrophy (MSA) is a neurodegenerative disorder associated with the degeneration of nerve cells in specific areas of the brain. This cell degeneration causes problems with movement, balance, and other autonomic functions of the body, such as bladder control or blood pressure regulation.

[0126] The cause of MSA is unknown, and no specific risk factors have been identified. Approximately 55% of cases occur in men, with the typical age of onset being in the late 50s to early 60s. MSA often presents with some of the same symptoms as Parkinson's disease. However, patients with MSA typically respond poorly, if at all, to dopaminergic medications used for Parkinson's disease.

[0127] N. frailtyFrailty syndrome ("frailty") is a geriatric syndrome characterized by functional and physical decline, including reduced mobility, muscle weakness, physical slowness, lack of stamina, low physical activity, malnutrition, and involuntary weight loss. Such decline is often associated with cognitive dysfunction and the consequences of diseases such as cancer. However, frailty can also occur in the absence of disease. Frail individuals are at increased risk for poor outcomes due to fractures, falls, disability, comorbidities, and premature death (C. Buigues, et al. Effect of a Prebiotic Formulation on Frailty Syndrome: A Randomized, Double-Blind Clinical Trial, Int. Mol. Sci. 2016, 17, 932). Furthermore, frail individuals are at increased risk for high medical costs (ibid.).

[0128] Common symptoms of frailty can be determined by certain types of tests. For example, unintentional weight loss involves a loss of at least 10 lbs or more than 5% of body weight in the previous year, muscle weakness can be determined by low grip strength in the bottom 20% (adjusted for gender and BMI) at baseline, physical slowness can be based on the time it takes to walk a 15-foot distance, lack of endurance can be determined by an individual's self-reported fatigue, and low physical activity can be measured using standardized questionnaires (Z. Palace et al., The Frailty Syndrome, Today's Geriatric Medicine 7(1), at 18(2014)).

[0129] In some embodiments, the methods and compositions are useful for slowing down the progression of age-related cognitive impairment. In other words, the cognitive ability of an individual declines more slowly after treatment with the methods of the present disclosure than before or without treatment with the methods of the present disclosure. In some such cases, the treatment method includes measuring the progression of cognitive decline after treatment and determining that the progression of cognitive decline is reduced. In some such cases, this determination is made by comparing with a reference subject, for example, the rate of cognitive decline in the individual before treatment, for example, determined by previously measuring cognition at two or more time points before administration of the blood product.

[0130] The present method and composition can also be used to stabilize the cognitive ability of individuals, such as individuals suffering from age-related cognitive decline or individuals at risk of suffering from age-related cognitive decline.For example, an individual may exhibit some age-related cognitive impairment, and the progression of cognitive impairment observed before treatment with the method of the present disclosure is stopped after treatment with the method of the present disclosure.As another example, an individual may be at risk of developing age-related cognitive decline (for example, the individual may be over 50 years old or may have been diagnosed with age-related disorders), and the cognitive ability of the individual is not substantially changed after treatment with the method of the present disclosure compared to before treatment with the method of the present disclosure.In other words, cognitive decline cannot be detected.

[0131] The present method and composition can also be used to reduce the cognitive impairment of individuals suffering from age-related cognitive impairment.In other words, the cognitive ability of individuals after treatment with the present method is improved.For example, the cognitive ability of individuals after treatment with the present method is improved by, for example, 2 times or more, 5 times or more, 10 times or more, 15 times or more, 20 times or more, 30 times or more, or 40 times or more, for example, 50 times or more, 60 times or more, 70 times or more, 80 times or more, 90 times or more, or 100 times or more, compared with the cognitive ability observed in individuals before treatment with the present method.In some cases, treatment with the present method and composition can restore the cognitive ability of individuals suffering from age-related cognitive decline to, for example, the level that the individuals had when they were about 40 years old or younger.In other words, cognitive impairment is suppressed.

[0132] The methods and blood products and fractions, including plasma, may also be used to treat undesirable conditions associated with post-surgical recovery and to promote post-surgical recovery. Such conditions and indications include, by way of example and not limitation, pain and wound healing. The methods and compositions of the present invention may also be used to treat acute pain and chronic pain in diseases or conditions not necessarily associated with post-surgical recovery. The methods and compositions may also be used to treat wound healing not necessarily associated with post-surgical recovery. The methods and compositions may also be used to promote or stimulate remyelination, treating diseases associated with myelination, such as multiple sclerosis.

[0133] The present methods and blood products and fractions, including plasma, can also be used to treat indications related to the nervous system. By way of example and not limitation, such conditions include central nervous system conditions such as central neuropathic pain, spinal cord injury, myelopathy, and central neuropathic pain associated with post-operative recovery. There are 170,000 new cases of spinal cord injury annually, with a prevalence of approximately 300,000, of which 40-75% present as central neuropathic pain (Jadad A et al., AHRQ Evidence Report Summaries, Agency for Healthcare Research and Quality; (1998-2005); word-wide-website: nscisc.uab.edu / Public / Facts%202016.pdf; world-wide-website: nscisc.uab.edu / PublicDocuments / fact_figures_docs / Facts%202012%20Feb%20Final.pdf). One-third of patients experience severe pain, and only one-third achieve a 50% or greater reduction in pain with treatment (Charbonneau R, CMAJ, 189(2): E48-E49 (2017); and Hadjipavlou G, et al., BJA Education, 16 (8): 264-68 (2016). The incidence of myelopathy is 605 per million, and although surgical options exist, there are no pharmacological treatments, indicating an unmet need in the field (Nouri A, et al., Spine, 40 (12): E675-93 (2015); The Lancet Neurology, editorial 18 (7): p615 (2019)).

[0134] By way of example, and not limitation, these disorders also include plexus / nerve root disorders, such as plexopathies, cervical radiculopathy, and sciatica (lumbar radiculopathy). The prevalence of plexopathies is 2-3 cases per 100,000 people. Current options include antiepileptic and antidepressant medications for neuropathic pain management, representing an unmet need. The prevalence of cervical radiculopathy is 100 cases per 100,000 men and 60 cases per 100,000 women (McCartney S, et al., Br.J.Gen.Pract.,68(666):44-46 (2018)). The annual prevalence of sciatica is 1-5%, and although many cases resolve spontaneously, sciatica responds less to treatment the longer the duration of the condition. Treatment options include surgical procedures, standard pain medications, and steroids, highlighting the need for new therapies (Lewis R et al., Health Technology Assessment - The Clinical Effectiveness and Cost-Effectiveness of Management Strategies for Sciatica: Systematic Review and Economic Model, No. 15.39 NIHR Journals Library (2011)).

[0135] Other indications include disorders of the peripheral nervous system. Non-limiting examples include peripheral neuropathy, peripheral neuropathy associated with postoperative recovery, carpal tunnel syndrome, chemotherapy-induced peripheral neuropathy, compression and trauma, diabetic neuropathy, peripheral neuropathy associated with shingles (postherpetic neuralgia), complex regional pain syndrome, and trigeminal neuralgia. Peripheral neuropathy is a disorder of the peripheral nerves that affects at least 20 million people in the United States. Approximately 60% of diabetic patients experience diabetic neuropathy, a type of peripheral neuropathy (see word-wide-website:healthcommunities.com / neuropathy / overview-of-neuropathy.shtml). Carpal tunnel syndrome affects 3-6% of adults, and treatments include splinting, steroids, and surgery (LelBlanc Ke, et al., Am Fam Physician, 83(8): 952-58 (2011)). Chemotherapy-induced peripheral neuropathy occurs in 40-60% of patients within three months of chemotherapy and up to three months after chemotherapy, affecting 650,000 patients annually. Peripheral neuropathy can lead to chemotherapy dose reduction or discontinuation and impact quality of life (QOL), but no medications or supplements have been shown to prevent this condition (JAMA Oncology, 5 (5): 750, (2019)). Compression and trauma-related peripheral neuropathy occurs in 2-3% of trauma patients, accounting for 3 million trauma cases in the United States. Surgery is often effective, but new pharmacological agents are needed (see American Association for the Surgery of Trauma - Trauma Facts, available at the word-wide-website: aast.org / trauma-facts; Novak CB, Medscape - Peripheral Nerve Injuries, (Oct. 5, 2018), available at https: / / emedicine.medscape.com / article / 1270360-overview).

[0136] An additional indication of the peripheral nervous system for which the present methods and blood products and fractions, including plasma, find therapeutic use is diabetic neuropathy. In the United States, the diabetic population is approximately 30 million, with neuropathy affecting 8-26% of patients (see Risson V, et al., Incidence and Prevalence of Painful Diabetic Neuropathy and Postherpetic Neuralgia in Major 5 European Countries, the United States and Japan, Value in Health (20): A339-A811 PSY18 (2017), available at the word-wide-website: valueinhealthjournal.com / article / S1098-3015(17)31179-8 / pdf). FDA-approved options for diabetic neuropathic pain include pregabalin, duloxetine, fluoxetine, and tapentadol, but not all of these medications are effective in many patients, and none of them directly address nerve damage.

[0137] Peripheral neuropathy associated with shingles (postherpetic neuralgia) may also be treated with the methods and blood products of the present invention. 20% of shingles patients experience postherpetic neuralgia, accounting for 1 million cases per year in the United States (see world-wide-website: emedicine.medscape.com / article / 1143066-overview#a6 word-wide-website: cdc.gov / shingles / hcp / clinical-overview.html). Gabapentin and pregabalin are approved for the treatment of this condition, but the pain is often difficult to treat (Sacks GM, Am J Manag Care 19 (1 Suppl): S207-13 (2013)).

[0138] Additional peripheral neuropathy indications, such as complex regional pain syndrome and trigeminal neuralgia, may be treated with the methods and compositions of the present invention. They occur in 5.5 to 26 cases per 100,000 population. They are associated with severe pain and disability, and responses to treatment are variable, representing a high unmet need. (See Complex Regional Pain Syndrome Fact Sheet, National Institutes of Health-National Institute of Neurological Disorders and Stroke, available at the following word-wide website: ninds.nih.gov / Disorders / Patient-Caregiver-Education / Fact-Sheets / Complex-Regional-Pain-Syndrome-Fact-Sheet.) Trigeminal neuralgia occurs in 4.2 to 28.9 cases per 100,000 population. It significantly impacts quality of life, becomes resistant to treatment over time, and requires patients to try many different medications (Wu N, et al., J Pain, 18 (Suppl 4): S69, (2017)). The only approved medication is carbamazepine. Therefore, there is an unmet need to treat the pain experienced by these patients.

[0139] Additional indications that can be treated with the methods and compositions of the present invention include central post-stroke pain, central pain in multiple sclerosis, post-traumatic headache, Dejerine-Roussy syndrome, optic neuritis, mitochondrial optic neuropathy, ischemic optic neuropathy, neuromyelitis, hereditary optic neuropathy, alcoholic neuropathy, Guillain-Barré syndrome, chronic inflammatory demyelinating polyneuropathy (CIDP), multifocal motor neuropathy (MNN), paraneoplastic autonomic neuropathy, peripheral neuropathy associated with sarcoidosis, peripheral neuropathy associated with rheumatoid arthritis, peripheral neuropathy associated with systemic lupus erythematosus, peripheral neuropathy associated with Sjogren's syndrome, peripheral neuropathy associated with celiac disease, Bell's palsy, and peripheral neuropathy associated with Lyme disease. Examples of peripheral neuropathy include peripheral neuropathy caused by hepatitis B, peripheral neuropathy associated with leprosy, peripheral neuropathy associated with hepatitis C, peripheral neuropathy associated with HIV / AIDS, peripheral neuropathy associated with amyloidosis, anti-MAG associated peripheral neuropathy, peripheral neuropathy associated with cryoglobulinemia, peripheral neuropathy associated with POEMS, toxin-induced peripheral neuropathy, peripheral neuropathy associated with kidney disease, peripheral neuropathy associated with vasculitis, peripheral neuropathy associated with vitamin and nutrient deficiencies, Charcot-Marie-Tooth disease (CMT), idiopathic peripheral neuropathy, fibromyalgia, and paraneoplastic peripheral neuropathy.

[0140] The present methods and blood products and fractions, including plasma, can also be used to treat indications related to wound healing. By way of example and not limitation, the wound may be an abrasion, an abrasion, an incision, a laceration, or a puncture wound. Such indications include both chronic and acute wounds. By way of example and not limitation, wound indications include acute wounds such as surgical wounds, trauma, and burns, as well as chronic wounds such as diabetic ulcers, pressure ulcers, venous ulcers, and arterial ulcers, although any type of chronic or acute wound can be treated by the methods and compositions of the present invention.

[0141] Diabetic ulcers affect more than 2.2 million people in the United States, representing 6.4% of the global incidence (Chun D, ​​et al., J Clin Med, 8:748 (2019)). Despite several treatment options, such as wound excision and medical dressings, many patients endure infection and ultimately require amputation, highlighting the need for new treatments, particularly pharmacological ones.

[0142] Pressure ulcers occur in 1.8% of all hospitalized patients, amounting to hundreds of thousands of cases annually (Bauer K, et al., Ostomy Wound Manage, 62 (11): 30-38 (2016)). As with diabetic ulcers, treatment options exist, such as wound debridement and medical dressings, but many patients experience infection, and ulcers can lead to death.

[0143] Venous ulcers, primarily occurring in the legs, place a significant burden on the elderly and affect approximately 1% of the global population (Nelzen O, Phlebolymphology, 15 (4)(2008)). Venous ulcers are difficult to heal and are more prone to recurrence than other chronic ulcers. As with diabetic and pressure ulcers, treatment options exist, such as wound debridement and medical dressings, but their recurrence highlights the need for new treatments, particularly pharmacological therapies. Arterial ulcers occur in approximately one-quarter of venous ulcers (see Gabriel A, Vascular Ulcers, (2018) available at https: / / emedicine.medscape.com / article / 1298345-overview#a6). Treatment options include wound debridement and medical dressings, but approved pharmacological agents are lacking.

[0144] Approximately 1.3 million patients develop surgical wounds annually (see MediWound - Innovating Solutions for Wound & Burn Care (2019) at 19 available at http: / / ir.mediwound.com / static-files / cd547017-d1ed-460e-8cb2-0550b1e18a29). Surgical wounds are cuts or incisions in the skin, typically made with a scalpel during surgery, but can also result from the placement of drains during surgery. Surgical wound healing is an important outcome of surgery. Postoperative wound disruption or separation of wound layers with fascial disruption can be a serious complication (see "Hospital Harm Improvement Resource-Wound Disruption" (2016) available at world-wide-website:patientsafetyinstitute.ca / en / toolsResources / Hospital-Harm-Measure / Documents / Resource-Library / HHIR%20Wound%20Disruption.pdf). Also, surgical wounds take significantly longer to heal in older patients than in younger patients (Gerstein AD, Dermatol Clin, 11(4): 749-57(1993)).

[0145] Traumatic wounds primarily include cuts, lacerations, punctures, and abrasions, which cause damage to the skin and underlying tissues. Traumatic wounds are typically categorized into three groups: acute wounds, incisions, and penetrating wounds. Acute wounds occur when the skin is torn or broken, are jagged in appearance, and usually contain foreign material such as glass, metal, gravel, sand, or dirt. Incisions occur when a sharp object penetrates the skin and underlying subcutaneous tissue. Penetrating wounds are the deepest and most severe of the three types. Stab wounds and gunshot wounds are typical examples. (See Traumatic Wounds, available on the Word-wide website: woundcarecenters.org / article / wound-types / traumatic-wounds and Leaper DJ, BMJ, 332 (7540): 532-35 (2006)). While some physical treatment options exist (e.g., sutures), the need for pharmacological intervention remains.

[0146] The World Health Organization (WHO) estimates that burns cause 180,000 deaths each year. Nonfatal burns are also a major cause of morbidity, including prolonged hospitalization (word-wide-website: who.int / news-room / fact-sheets / detail / burns). Typical treatments include surgical management and bandaging. Pharmacological treatments focus on analgesia, infection control, sedation, volume repletion, anticoagulation, and nutrition (Green A, et al., Clinical Pharmacist, 2:249-54 (2010)). The methods and compositions of the present invention may fulfill an unmet need for pharmacological interventions that promote healing of injuries to the skin and subcutaneous tissue.

[0147] The present methods and blood products and fractions, including plasma, can be used to treat conditions and indications associated with post-operative recovery at different times. By way of example and not limitation, administration to a subject can occur pre-operatively, peri-operatively (during the procedure), or post-operatively.

[0148] In one embodiment of the present invention, the method and blood products and fractions, including plasma, can be used to treat pain. By way of example and not limitation, such pain can include acute pain or chronic pain. In other embodiments of the present invention, the method and blood products and fractions, including plasma, can also be used to treat central pain or central neurological disorders. Central pain includes neurological conditions caused by damage or dysfunction of the central nervous system (CNS), such as the brain, brainstem, or spinal cord. It can affect most of the body or be limited to a specific area. The pain can be constant or intermittent. The intensity of the pain can be moderate to severe. Such pain can also be affected by touch, movement, emotion, and temperature changes. Pain may develop immediately after the inciting event or may be delayed for months or years (Central Pain Information Page - National Institute of Neurological Disorders and Stroke, Central Pain Syndrome Information Page is available on the Word-wide website: ninds.nih.gov / disorders / all-disorders / central-pain-syndrome-information-page; see Colloca L, et al., Nat Rev Dis Primers, 3:17002 (2017)). Further embodiments of the present invention include using the methods and blood products and fractions, including plasma, to treat spinal cord injury (SCI), myelopathy, neuropathy, plexopathy, cervical radiculopathy, sciatica (lumbar radiculopathy); central post-stroke pain, central pain in multiple sclerosis, post-traumatic headache, Dejerine-Roussy syndrome, optic neuritis, mitochondrial optic neuropathy, ischemic optic neuropathy, neuromyelitis optica, and hereditary optic neuropathies.

[0149] Another embodiment of the present invention is that the methods and blood products and fractions, including plasma, may also be used to treat peripheral pain or peripheral neuropathy. Peripheral neuropathy can refer to several conditions involving damage to the peripheral nervous system. Over 100 peripheral neuropathies have been identified, depending on which type of nerve is damaged, including motor, sensory, autonomic, etc. (Central Page Information Page - National Institute of Neurological Disorders and Stroke, Peripheral Neuropathy Fact Sheet available on the Word-wide website: ninds.nih.gov / Disorders / Patient-Caregiver-Education / Fact-Sheets / Peripheral-Neuropathy-Fact-Sheet; and Colloca L, et al., Nat Rev Dis Primers, 3:17002 (2017)).Further embodiments of the present invention are directed to the treatment of peripheral neuropathy associated with carpal tunnel syndrome, chemically induced peripheral neuropathy, compression and trauma, diabetic neuropathy, peripheral neuropathy associated with shingles (postherpetic neuralgia), complex regional pain syndrome, trigeminal neuralgia, alcoholic neuropathy, Guillain-Barre syndrome, chronic inflammatory demyelinating polyneuropathy (CIDP), multifocal motor neuropathy (MNN), paraneoplastic autonomic neuropathy, peripheral neuropathy associated with sarcoidosis, peripheral neuropathy associated with rheumatoid arthritis, peripheral neuropathy associated with systemic lupus erythematosus, peripheral neuropathy associated with Sjogren's syndrome, peripheral neuropathy associated with celiac disease, Bell's palsy, peripheral neuropathy associated with Lyme disease, peripheral neuropathy associated with leprosy, B The present invention also includes using the methods and blood products and fractions, including plasma, to treat peripheral neuropathy associated with hepatitis B, hepatitis C, HIV / AIDS, amyloidosis, anti-MAG, cryoglobulinemia, POEMS, toxin-induced peripheral neuropathy, renal disease, vasculitis, vitamin and nutrient deficiency, Charcot-Marie-Tooth disease (CMT), idiopathic peripheral neuropathy, and fibromyalgia and paraneoplastic peripheral neuropathy.

[0150] One embodiment of the present invention is that the method and blood products and fractions, including plasma, can be used to treat wounds by promoting wound healing. Other embodiments of the present invention include using the method and blood products and fractions, including plasma, to treat chronic or acute wounds. Still other embodiments of the present invention include treating diabetic ulcers, pressure ulcers, venous ulcers, arterial ulcers, surgical wounds, and traumatic wounds.

[0151] 14. Methods for diagnosing and monitoring improvement in neurocognitive-related disorders In some cases, among various methods for diagnosing neurocognitive-related diseases and monitoring their progression and improvement, the following types of assessments are used alone or, if desired, in combination with an individual suffering from a neurodegenerative disease. The following types of methods are provided as examples and are not limited to the methods listed. If desired, any convenient method for monitoring disease may be used in the practice of the present invention. These methods are also contemplated by the methods of the present invention.

[0152] A. Global cognition Method embodiments of the present invention further include methods for monitoring the effects of a drug or treatment on a subject for the treatment of cognitive impairment and / or age-related dementia, comprising comparing cognitive function before and after treatment. Those skilled in the art will recognize that there are well-known methods for assessing cognitive function. For example, and without limitation, the methods may include assessment of cognitive function based on medical history, family history, physical and neurological examinations by clinicians specializing in dementia and cognitive function, laboratory tests, and neuropsychological evaluation. Further embodiments contemplated by the present invention include assessment of state of consciousness using, for example, the Glasgow Coma Scale (EMV), mental status tests including the Abbreviated Mental Test Score (AMTS) or the Mini-Mental State Examination (MMSE) (Folstein et al., J. Psychiatr. Res 1975;12:1289-198), global assessment of higher-order function, and estimation of intracranial pressure by funduscopy, etc.

[0153] In one embodiment, peripheral nervous system testing can be used to assess cognitive function, including smell, visual field and vision, eye movements and pupils (sympathetic and parasympathetic), facial sensory function, strength of the muscles of the face and shoulder girdle, hearing, taste, pharyngeal movements and reflexes, tongue movements (which can be tested individually (e.g., visual acuity can be tested with a Snellen eye chart, and reflexes including the masseter, biceps, and triceps tendons, hamstrings, Achilles tendon reflex, and plantar reflex (i.e., Babinski sign) are tested with the use of a reflex hammer)), muscle strength, muscle tone, and signs of rigidity, frequently on an MRC scale of 1-5.

[0154] 15. Reagents, Devices, and Kits Also provided are reagents, devices, and kits thereof for practicing one or more of the above methods. The reagents, devices, and kits thereof can vary widely.

[0155] Reagents and devices of interest include, for example, anticoagulants, cryopreservatives, buffers, isotonic solutions, etc., as described above with respect to the methods of preparing blood products, including plasma, for transfusion into a subject in need thereof.

[0156] The kit may also include blood collection bags, tubing, needles, centrifuge tubes, etc. In yet other embodiments, the kits described herein include two or more containers of plasma product, such as plasma protein fractions, e.g., three or more, four or more, five or more, e.g., six or more containers of plasma product. In some cases, the number of individual containers of plasma product in the kit may be nine or more, twelve or more, fifteen or more, eighteen or more, twenty-one or more, twenty-four or more, thirty or more, e.g., thirty-six or more, e.g., forty-eight or more. Each container may be associated with identifying information that includes various data regarding the plasma product contained therein, which may include one or more of the age of the donor of the plasma product, processing details regarding the plasma product, e.g., whether the plasma product has been processed to remove proteins above an average molecular weight (e.g., those described above), blood type details, etc. In some cases, each container in the kit includes identifying information for the plasma contained therein, including information regarding the donor age of the plasma product, e.g., the identifying information identifies data regarding the age of the plasma product donor (where such identifying information can be the donor's age at the time of collection). In some cases, each container in the kit contains plasma product derived from a donor of substantially the same age. That is, all of the containers contain product derived from donors of substantially, if not exactly the same, age. Substantially the same age means that the various donors from whom the plasma products in the kit are obtained differ from each other, in some cases, by 5 years or less, e.g., 4 years or less, e.g., 3 years or less, e.g., 2 years or less, e.g., 1 year or less, e.g., 9 months or less, 6 months or less, 3 months or less, e.g., 1 month or less. The identifying information may be present on any convenient container component, e.g., a label, RFID chip, etc. The identifying information may be human-readable, computer-readable, etc., as desired. The container may have any convenient configuration. The volume of the container can vary, but in some cases the volume ranges from 10 mL to 5000 mL, e.g., 25 mL to 2500 mL, e.g., 50 mL to 1000 mL, e.g., 100 mL to 500 mL. The container can be rigid or flexible and can be made from any convenient material, e.g., a polymeric material, including a medical-grade plastic material. In some cases, the container has a bag or pouch configuration.In addition to the container, such kits may further include an administration device, e.g., as described above. The components of such kits may be provided in any suitable packaging, e.g., a box or similar structure configured to contain the container and other kit components.

[0157] In addition to the above components, the kit further includes instructions for practicing the method. These instructions may be present in the kit in various forms, one or more of which may be present in the kit. One form in which these instructions may be present is information printed on a suitable medium or substrate, such as a sheet or sheets of paper on which the information is printed, in the kit's packaging, in a package insert, etc. Yet another means is a computer-readable medium on which the information is recorded, such as a diskette, CD, portable flash drive, etc. Yet another means may be a website address that can be used to access information on a remote site via the Internet. Any convenient means may be present in the kit.

[0158] 16. Experimental Procedure The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the present invention, and are not intended to represent that the experiments below represent all or the only experiments performed. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperatures, etc.), but some experimental error and deviation should be accounted for. Unless otherwise indicated, parts are parts by weight, molecular weight is weight average molecular weight, temperature is in degrees Celsius, and pressure is near atmospheric.

[0159] General methods in molecular and cellular biochemistry are described in Molecular Cloning: A Laboratory Manual, 3rd Ed. (Sambrook et al., Harvard Laboratory Press 2001), Short Protocols in Molecular Biology, 4th Ed. (Ausubel et al. eds., John Wiley & Sons 1999), Protein Methods (Bollag et al., John Wiley & Sons 1996), Nonviral Vectors for Gene Therapy (Wagner et al. eds., Academic Press 1999), Viral Vectors (Kaplift & Loewy eds., Academic Press 1995), Immunology Methods Manual (I. Lefkovits ed., Academic Press 1997), and Cell and Tissue Culture: Laboratory Precedures in Biotechnology (Doyle & Griffiths, John Wiley & Sons Genetic engineering techniques, such as those described in the "Genetic Engineering of Genes," can be found in standard textbooks, such as "Genetic Engineering of Genes," ...

[0160] Example 1 - Treatment of age-related cognitive disorders A. Materials and Reagents USP saline was purchased from Hospira (Lake Forest, IL). Injections were performed using a 27.5G or 30G needle, with a volume of 150 μL per injection. Commercially available PPF ("PPF1"), such as the commercial PPF preparation described above in a 5% solution, was stored at 4° C. Commercially available HAS ("HAS1"), such as the commercial HAS preparation described above in a 5% solution, was stored at 4° C.

[0161] B. Animal Supply and Housing The mouse strains NOD.CB17-Prkdcscid / NcrCrl ("NODscid," strain code 394, Charles River, MA) (Bosma, M. et al., The scid mouse mutant. 137 Curr Top Microbiol Immunol 197 (1988)) and NODscid gamma ("NSG," strain code 005557, The Jackson Laboratory, Bar Harbor, ME) were used. Each mouse was ear-punched and assigned a unique identification number. All mice were individually housed under specific pathogen-free conditions with a 12-hour light / 12-hour dark cycle. All animal handling and use followed standard IACUC-approved guidelines.

[0162] C. Administration Unless otherwise stated below, NSG and NODscid mice were injected intravenously via tail vein injection (150 μL per injection) with USP saline, 5% PPF1, or 5% HAS1 twice a week for up to 6 months.

[0163] D. Open field The open field test was used to assess the exploratory behavior of subject mice. The open field test is an empty testing arena, usually circular or square. Mice are placed in a 50 cm x 50 cm open field arena for 15 minutes, and their activity level is measured. Rearing time was measured by tracking the duration the forelimbs remained in contact with the walls of the box. Total distance covered and speed were also measured throughout the test period. A CleverSys TopScan V3.0 (Reston, VA) was used to track the behavior of mice in the open field. The open field chamber was constructed by CleverSys.

[0164] EY Maze Mice were allowed to explore two arms (start + non-novel) of the Y-maze for 5 min. After 1 h, mice were allowed to explore all three arms and the total time and number of entries into these arms were recorded.

[0165] F.Barnes Maze Mice were trained for four consecutive days in a modified Barnes maze, with a maximum of 120 seconds to find the exit hole (Barnes, CA, Memory deficits associated with senescence: A neurophysiological and behavioral study in the rat, J. COMPARATIVE AND PHYSIOLOGICAL PSYCHOLOGY, 93(1):74-104 (1979); see also Faizi, M. et al., The Thy1-hAPP (Lond / Swe+) mouse model of Alzheimer's disease displays broad behavioral deficits in sensorimotor, cognitive, and social function., BRAIN BEHAV. 2(2):142-54 (2012) for a description of the modified maze). The exit hole remained the same for the four trials on a training day but was changed between training days. The latency to find the exit hole was recorded for each cohort of mice on each of the four training days.

[0166] G. DCX-positive cells and Ki67-positive cells Doublecortin (DCX) is a microtubule-associated protein expressed by neural progenitor cells and also by immature neurons within embryonic and adult cortical structures. Neural progenitor cells express DCX when they are actively dividing. The protein is downregulated after two weeks. Because of this association, it is useful as a marker of neurogenesis.

[0167] Brain tissue processing and immunohistochemistry were performed on free-floating sections using well-described techniques (Luo, J. et al. Glia-dependent TGF-β signaling, acting independently of the TH17 pathway, is critical for the initiation of murine autoimmune encephalomyelitis. J. CLIN. INVEST. 117, 3306-3315 (2007)). Mice were anesthetized and perfused with 0.9% saline. Brains were removed and fixed in 4% paraformaldehyde-phosphate buffer at pH 7.4 at 4°C, then submerged in 30% sucrose for cryoprotection. Brains were then sectioned at 30 μm using a cryomicrotome at -22°C. Sections were stored in cryoprotective medium. The primary antibodies used were goat anti-Dcx (Santa Cruz Biotechnology, 1:500 for twice-weekly dosing experiments or 1:200 for three-weekly dosing experiments) or rabbit anti-Ki67 (1:500, Abcam). Primary antibody staining was revealed using diaminobenzidine (DAB, Sigma-Aldrich) or fluorescently conjugated secondary antibodies, followed by biotinylated secondary antibodies and the ABCkit (Vector). To estimate the total number of Dcx-positive cells per dentate gyrus, immunopositive cells within the granule cell and subgranule cell layers of the dentate gyrus were counted and averaged in three coronal brain sections through the hippocampus.

[0168] H. Barnes maze testing of aged NSG mice treated with young plasma, effluent I, or effluent II / III. Aged NSG mice (12 months old) were divided into groups (n = 14 in total) and given 150 μL of saline, young plasma, effluent I, or effluent II / III via tail vein injection before the start of behavioral testing. Each separate group was divided into three cohorts, and behavioral testing began in different weeks for each cohort.

[0169] I. Barnes maze and cell viability (BrdU staining) of aged NSG mice treated with young plasma or PPF1 three times a week. Aged (12-month-old) male NSG mice were treated intravenously with 150 μL of purified young human plasma (young plasma), PPF1, or saline via tail vein injection three times per week for four weeks. This regimen was changed to twice per week during weeks 5 and 6, which were the behavioral testing weeks.

[0170] Prior to treatment, mice were divided into three cohorts of 13–15 mice each, and each cohort received intraperitoneal (ip) BrdU injections 5 days prior to the initiation of treatment with young plasma, PPF1, or saline.

[0171] During weeks 5 and 6, behavioral testing was conducted to measure the latency of each mouse to reach the target hole in a Barnes maze test. Each test session lasted a maximum of 120 seconds. Finding the target hole was recorded using software that determined when the mouse's nose entered the area defined as the target hole.

[0172] At the end of behavioral testing, animals were sacrificed and six sections per hippocampus were quantified using bright-field microscopy to determine the presence of BrdU-positive cells within the granule cell layer of the dentate gyrus. To obtain an estimate of the total number of BrdU-positive cells, the average number of BrdU-positive cells was multiplied by 72, the total number of sections per hippocampus for each animal, for representative sections across different regions of the hippocampus.

[0173] J. Neurosphere and Cortical Culture Assays 1. Tuj1 and DAPI staining Mouse C57 E14,15 cortices (Lonza:M-CX-300) were suspended in 12 mL of neurobasal medium supplemented with B27 and 2 mM Glutamax (Sigma-Aldrich). 200 μL was added to each well of a 96-well plate pre-coated with collagen I (Corning, Inc.). After 16 hours, the seeding medium was replaced with pre-warmed (37°C) control medium (neurobasal medium supplemented with B27 and 2 mM Glutamax (Gibco)). On day 4 of in vitro culture ("days in vitro" or "DIV"), the culture medium was replaced with fresh control medium, control medium and 10% PPF1, control medium and 10% HAS1, vehicle and 10% PPF1, or vehicle and 10% HAS1. Cultures were maintained for 21 days, with 75% of the medium replaced with fresh medium every 3 days. At 21 DIV, cultures were washed three times with PBS and then fixed with 4% paraformaldehyde for 20 minutes at room temperature (RT). After fixation, cultures were washed twice with PBS and then permeabilized with 0.1% Triton X100 for 5–20 minutes. After permeabilization, cultures were blocked with 3% bovine serum albumin (Sigma-Aldrich) for 60 minutes at RT. After 60 minutes, the blocking solution was aspirated, and cultures were labeled with anti-Tuj1 antibody (AbCam-1:500) overnight at 4°C. After labeling, cultures were washed three times with PBS + 0.1% BSA and then stained with A647-conjugated donkey anti-mouse antibody (1:1000) overnight at 4°C. Cultures were then washed twice with PBS and labeled with Hoechst 33342 (1:1000) for 20 minutes. After Hoechst labeling, the samples were washed three times with PBS. 25 fields were acquired for each well using a GE InCell Analyzer 2000 (GE Healthcare Life Sciences) at 10x magnification. The results are shown in Figure 19.

[0174] 2. Net neurite length Net neurite length was determined from the cultures described in the previous section. Neurite analysis was performed using a custom algorithm created with the GE InCell Investigator Developer Toolbox. Results from control and vehicle-treated samples were nearly identical and were combined for statistical analysis. Results are shown in Figure 20.

[0175] 3. Number and size of cortical spheres, length of processes, and branching Mouse C57 E14,15 cortices (Lonza:M-CX-300) were suspended in 12 mL of neurobasal medium supplemented with B27 and 2 mM Glutamax (Sigma-Aldrich). 200 μL was added to each well of a 96-well plate pre-coated with polylysine and laminin. After 4 days, 50% of the medium was replaced with fresh medium, and cells were treated with the test article (vehicle, PPF1, or HAS1) to a final concentration of 10%. This process was repeated 3 days later. On day 7 of treatment, phase-contrast images of cells were taken at 10x magnification using an IncuCyte (Ann Arbor, MI) and analyzed using the standard "Neurite and Cell-Body" algorithm. Six replicates were analyzed, with four images taken per replicate. Standard errors are shown. Significance is indicated as P<0.5 using a two-tailed t-test. Results are shown in Figures 21 and 22.

[0176] 4. Staining Neurospheres with Sox2 Mouse C57 E14,15 cortical neurons (Lonza:M-CX-300) were suspended at 100–200K cells / mL in neurobasal medium supplemented with B27 and 2 mM Glutamax (Sigma-Aldrich). 200 μL was added to each well of a 96-well plate pre-coated with collagen I (Corning, Inc.). After 16 h, the seeding medium was replaced with pre-warmed (37°C) control medium (neurobasal medium supplemented with B27 and 2 mM Glutamax (Gibco)). On day 4 of in vitro culture ("days in vitro" or "DIV"), the culture medium was replaced with fresh control medium, control medium containing HAS vehicle (vehicle), control medium plus 10% PPF1, or control medium plus 10% HAS1. Cultures were maintained for 21 days, with 75% of the medium replaced with fresh medium every 3–4 days. At 21 DIV, cultures were washed three times with PBS and then fixed with 4% paraformaldehyde for 20 minutes at room temperature (RT). After fixation, cultures were washed twice with PBS and then permeabilized with 0.1% Triton 100X for 5–20 minutes. After permeabilization, cultures were blocked with 3% bovine serum albumin (Sigma-Aldrich) for 60 minutes at RT. After 60 minutes, the blocking solution was aspirated, and cultures were labeled with anti-Tuj1 antibody (AbCam - 1:500) and rabbit anti-SOX2 (AbCam: 1:5000) overnight at 4°C. After labeling, cultures were washed three times with PBS + 0.1% BSA and then stained with donkey anti-mouse 647 (AbCam) and sheep anti-rabbit Texas Red (1:1000) overnight at 4°C. Cultures were then washed twice with PBS and labeled with Hoechst (1:1000) for 20 minutes. After Hoechst labeling, samples were washed three times with PBS. Using 10x magnification on an InCell Analyzer 2000 (GE Healthcare Life Sciences), 20 or 25 fields were acquired for each well. Analysis of neurospheres and neurites was performed using custom algorithms created with the GE InCell Investigator Developer Toolbox. Results from control and vehicle-treated samples were nearly identical and therefore combined for statistical analysis.The results are shown in FIG.

[0177] K. In vivo experimental results 1. Open field test using 3-month-old and 13-month-old NSG mice NSG mice aged 3 months (young) or 13 months (old) were placed in an open field chamber for 15 minutes. Rearing time (Figure 1), speed (Figure 2), and distance (Figure 3) were measured. Figure 1 shows that 13-month-old mice had less rearing time than 3-month-old mice, but PPF1- and HAS1-treated mice were not significantly different from young mice. Figure 2 shows that 13-month-old saline-treated (control) and PPF1-treated mice were significantly slower than 3-month-old mice. However, HAS1-treated mice were significantly faster than saline-treated mice and were not significantly different from young mice. Figure 3 shows that aged saline-treated (control) and HAS1-treated mice had lower locomotor activity than young mice, and PPF1-treated mice covered a longer distance than saline-treated mice. All data shown are mean ± sem. * P<0.05, ** P<0.01, *** P<0.001, t-test, n=20, 18, 18, 19 (SAL=saline).

[0178] 2. Y-maze test using 3-month-old and 13-month-old NSG mice Young (3-month-old) and aged (13-month-old) NSG mice were tested in a cued Y-maze as a test of memory. Figure 4 shows that all mice spent significantly more time in the novel (N) arm than the non-novel (F) arm. Figure 5 shows that HAS1-treated aged mice had significantly impaired memory for the non-novel arm compared to young mice, while PPF1-treated mice tended to have improved memory for the non-novel arm. Figure 6 shows that aged saline- and PPF1-treated mice were significantly slower than young mice, but HAS1-treated mice were not significantly different from young mice. Figure 7 shows that aged saline- and PPF1-treated mice covered shorter distances than young mice, but HAS1-treated mice were not significantly different from young mice. All data shown are mean ± sem. * P<0.05, ** P<0.01, *** *P<0.001, paired t test, n=20, 18, 18, 19 (SAL=saline).

[0179] 3. Fear conditioning test for memory using 3-month-old and 13-month-old NSG mice Young (3-month-old) and aged (13-month-old) NSG mice were tested in a fear conditioning memory test. Figure 8A shows that 13-month-old mice tended to freeze for shorter periods of time than 3-month-old mice, while HAS1-treated mice freezed for approximately the same amount of time as 3-month-old mice. Figure 8B shows that 13-month-old control-treated mice performed poorly and freezed for the shortest periods of time in the auditory cue memory test. HAS1-treated mice tended to freeze for longer periods of time, indicating improved memory for the sound. Figure 9 shows quantification of the final 90 seconds of the memory cue test, showing that HAS1-treated mice tended to freeze for longer periods of time, indicating improved memory. n = 20, 16, 17, 19 (SAL = saline).

[0180] 4. Barnes maze test for spatial memory using 3-month-old and 13-month-old NSG mice Young (3-month-old) and aged (13-month-old) NSG mice were tested in the Barnes maze test for spatial memory. Figure 10A shows that 3-month-old mice performed best, taking the shortest time to reach the target hole by the last trial. Figure 10B shows quantification of the average of the last three trials, indicating that target hole memory was significantly impaired in aged saline- and HAS1-treated mice compared with young mice, but not in PPF1-treated mice. ** P<0.01, *** P<0.001, unpaired t-test, n=20, 18, 18, 19 (SAL=saline).

[0181] 5. Immunostaining using 3-month-old and 13-month-old NSG mice Brain sections were stained for doublecortin (Dcx), a marker of newborn neurons, and Ki67, a marker of proliferating cells, from 3- and 13-month-old NSG mice treated twice weekly with saline, PPF1, or HAS1. Dcx- and Ki67-positive cells were counted in the dentate gyrus of young and aged NSG mice. Figures 11A and 11B show that all aged mice had dramatically lower numbers of Dcx- and Ki67-positive cells, respectively. PPF1- and HAS1-treated mice tended to have increased numbers of Dcx- and Ki67-positive cells compared with saline-treated mice.

[0182] 6. Immunostaining using 3-month-old and 13-month-old NSG mice treated with PPF1 and HAS1 three times a week Brain sections from 13-month-old mice were stained for doublecortin (Dcx), a marker of newborn neurons, and Ki67, a marker of proliferating cells. Mice were treated with saline, PPF1, 1x concentrated HAS1, or 5x concentrated HAS1 three times per week. Dcx- and Ki67-positive cells were counted in the dentate gyrus. Figure 12 shows that mice treated with PPF1 tended to have increased neurogenesis (as indicated by Dcx staining) compared with saline-treated control animals. It also shows that more concentrated HAS1 tended to increase neurogenesis compared with saline-treated animals.

[0183] Figure 13 shows that cell proliferation (as indicated by Ki67 staining) was significantly increased in mice treated with PPF1 compared to saline-treated control animals, and that more concentrated HAS1 tended to increase neurogenesis compared to saline-treated animals. * P<0.05, unpaired t-test versus saline group, all data shown are mean±sem.

[0184] 7. Open field test with NODscid mice NODscid mice were treated intravenously with either saline or PPF1 via tail vein injection twice weekly starting at 6 months of age. Each group initially contained 20 mice. Mice were placed in an open field chamber for 15 minutes, and locomotor activity was recorded. Figure 14A shows that PPF1-treated mice tended to have increased rearing activity compared to saline-treated mice. Figures 14B and 14C show that PPF1-treated mice also tended to have improved speed and distance covered, respectively, compared to saline-treated mice.

[0185] 8. Barnes maze using aged (12-month-old) NSG mice treated with young plasma, effluent I, and effluent II / III Aged NSG mice (12 months old) were divided into groups (all with a size of n = 14) and received 150 μL of saline, young plasma, effluent I, or effluent II / III via tail vein injection before the start of behavioral testing. Each separate group was further divided into three cohorts, and behavioral testing began in each cohort on a different week. Mice were tested in a modified Barnes maze (as described above) to assess spatial learning and memory. Figure 15 shows that treatment with young plasma, effluent I, or effluent II / III tended to significantly improve the latency of aged NSG mice to reach the target hole.

[0186] 9. Barnes maze and cell viability using aged NSG mice treated with young plasma and PPF1 As described above, aged male NSG mice (12 months old) were treated with 150 μL of purified young human plasma (young plasma), PPF1, or saline three times per week (iv) for 4 weeks and twice per week during weeks 5 and 6, the weeks in which the reported studies were performed.

[0187] Figure 16 shows the latency to reach the hole in the Barnes maze for each treatment cohort. Treatment with PPF1 significantly improved spatial memory in aged mice compared to controls, and treatment with young plasma tended to improve spatial memory compared to controls (n: saline = 12, PPF1 = 14, young plasma = 11). * P<0.05, mean±sem, unpaired T-test.

[0188] Figure 17 shows the mean latency to find the target hole in the last three trials of each test day. Again, treatment with PPF1 significantly improved spatial memory in aged mice compared to controls, and treatment with young plasma tended to improve spatial memory compared to controls. * P<0.05, mean±sem, unpaired T-test.

[0189] Figure 18 shows the effect of young human plasma and PPF1 on cell viability, as determined by the number of BrdU-positively labeled cells (i.e., proliferating cells) within the granular layer of the dentate gyrus in aged (12-month-old) NSG mice. BrdU was administered (ip) for 5 days prior to the start of intravenous injections of young plasma, PPF1, or saline control, as described above. A significant increase in cell viability was observed in both young human plasma- and PPF1-treated mice compared to saline control. Statistical significance was determined using one-way ANOVA with Dunnett's multiple comparison post-hoc analysis between PPF1 and young human plasma compared to saline treatment (n: saline = 13, PPF1 = 13, young plasma = 11, **** P>0.0001, unpaired T-test between PPF1 or young human plasma and saline treatment).

[0190] L. In vitro neurosphere and cortical culture assay results Figure 19 shows that PPF1 and HAS1 differentially regulate neurosphere proliferation in cortical cultures. E14-15 C57 mouse cortices were cultured on collagen I-coated 96-well plates in culture medium containing vehicle alone, PPF1 (10%), or HAS1 (10%). Example images of neurospheres in cortical cultures after 21 days of in vitro culture, imaged for Tuj1 (neuron-specific class III beta-tubulin), DAPI (4',6-diamidino-2-phenylindole), or both Tuj1 and DAPI, are shown. Figure 19 shows that PPF1 increases the amount of neurospheres expressing either Tuj1 or DAPI. The increased Tuj1 expression indicates that PPF1-treated cortical cultures produce more neurospheres that differentiate into a more neuronal-like phenotype.

[0191] Figure 20 shows three cultures of C57 mouse E14-15 cortical neurons (Lonza:M-CX-300) suspended at 100-200K cells / mL in neurobasal medium supplemented with B27, 2 mM Glutamax (Sigma-Aldrich), coated on collagen I-coated 96-well plates in culture medium containing vehicle, PPF1 (10%), or HAS1 (10%). Net neurite length, indicative of neurogenesis, was increased in PPF1-treated cultures compared to control or HAS1-treated cultures.

[0192] Figure 21 shows three cultures of E14-15 cortical neurons (Lonza:M-CX-300) from C57 mice, suspended at 100-200K cells / mL in neurobasal medium supplemented with B27 and 2 mM Glutamax (Sigma-Aldrich) coated on collagen I-coated 96-well plates in culture medium containing vehicle, PPF1 (10%), or HAS1 (10%). The IncuCyte software algorithm, available from Essen BioSciences (Ann Arbor, MI), detected cortical culture spheres (highlighted in yellow) and processes (highlighted in pink). More spheres and processes were observed in PPF1-treated cultures, and increased sphere size and process branching were also observed in PPF1-treated cultures. Scale bars are 300 μm.

[0193] Figures 22A-D show sphere number, process length, process branch points, and sphere size, respectively. Quantification was performed using the IncuCyte software algorithm available from Essen BioSciences (Ann Arbor, MI). Standard errors are shown. Significance using a two-tailed t-test is indicated. Figure 22A shows that PPF1-treated cultures had an increased number of spheres compared to vehicle- or HAS1-treated cultures (P=0.0006, PPF1 vs. vehicle; P=0.0007, PPF1 vs. HAS1). Figure 22B shows that PPF1-treated cultures displayed increased process length compared to vehicle- or HAS1-treated cultures (P=4e -8, PPF1 vs. vehicle, P=0.002, PPF1 vs. HAS1, and P=0.018, HAS1 vs. vehicle). Figure 22C shows that PPF1-treated cultures produced more process branch points compared to vehicle- or HAS1-treated cultures (P=0.002, PPF1 vs. vehicle, P=0.004, PPF1 vs. HAS1). Figure 22D shows that PPF1-treated cultures were associated with increased sphere size compared to vehicle- or HAS1-treated cultures (P=0.002, PPF1 vs. vehicle, P=0.004, PPF1 vs. HAS1). Collectively, the results of this data indicate that treatment with PPF1 (and, to a lesser extent, treatment with HAS1) is associated with features indicative of increased cell growth and process formation in cortical cultures.

[0194] Figure 23 shows the number of neurospheres staining positive for Sox2, a transcription factor that plays an important role in the maintenance of embryonic and neural stem cells. Quantification was performed using an algorithm in the GE InCell Investigator Toolbox. PPF1-treated cultures significantly increased the number of neurospheres staining positive for Sox2, indicating that PPF1 treatment is associated with an increase in the number of cells with neurogenic potential.

[0195] Example 2 - Pain relief and post-operative recovery 1. Pain Model a) Pain - Pre-injury treatment (1) Neuropathic nerve damage changes A chronic pain model using chronic constriction injury (CCI) was used to determine the level of pain experienced by 22-month-old C57BL / 6J mice treated with (1) PPF1 after CCI, (2) vehicle after CCI surgery, or (3) vehicle after sham surgery. Using this model, the nervous system is conditioned into a persistent state of high reactivity that lowers the pain threshold long after the initial injury occurs (see, e.g., Safakhah, HA et al., Journal of Pain, 10:1457-66 and Suter MR, et al., Anesthesiology Res and Practice (2011), both of which are incorporated herein by reference in their entirety). Sciatic nerve

[0196] As measured by electrophoresis, PPF1 is a PPF containing approximately 88% normal human albumin (based on total protein), 12% alpha and beta globulins, and less than 1% gamma globulins. Unless otherwise specified, PPF1 is administered in vivo as a 5% solution (w / v, 50 g / L). PPF2 is also a PPF but differs significantly from PPF1. PPF2 meets the same protein content and concentration specifications as PPF1.

[0197] Figure 24 shows the timeline of the CCI experiment. 23-month-old wild-type mice underwent CCI surgery or sham surgery by ligation 24 hours before receiving either PPF1 or vehicle control at 150 μL / day (intravenously in the tail vein) in a 7-day continuous pulse-dose regimen. Behavior was assessed at 4 weeks, and tissue collection for histological examination was performed at 5 weeks.

[0198] Figure 25 shows the location of CCI surgery performed on a 23-month-old wild-type mouse. Ligatures were placed on the sciatic nerve as shown. Figure adapted from Suter MR et al., Anesthesiology Res and Practice (2011), incorporated herein by reference in its entirety.

[0199] Figure 26 shows data from the von Frey mechanical allodynia test for wild-type mice 4 weeks after CCI surgery or sham surgery, as detailed in Figure 24. To determine the animals' tolerance to mechanical pressure, the hind paws were stimulated with von Frey filaments of different thicknesses via the subject's sciatic nerve. The pressure with which the mice withdrew their hind paws was measured and plotted in Figure 26. The figure shows that mice treated with PPF1 after CCI surgery were significantly less painful (able to tolerate greater pressure) than mice treated with vehicle control after CCI surgery. Sham-operated animals also showed significantly less pain than mice treated with vehicle control after CCI surgery. The main finding is that PPF1 has a positive effect on the mechanical nociceptive deficit induced by CCI surgery. *** P<0.001 PPF1-treated CCI vs. vehicle-treated CCI; * *P<0.05 Sham-operated vehicle vs. CCI vehicle; one-way ANOVA with Tukey post-hoc analysis.

[0200] Figure 27 shows data from histological examination of the hippocampus performed on wild-type mice described in Figure 24. Neurogenesis was measured using the doublecortin (DCX) marker. Mice treated with PPF1 after CCI surgery had significantly more neurogenesis in the dentate gyrus of the hippocampus than mice treated with vehicle. Although both groups were treated with vehicle after surgery, sham-operated mice tended to have greater neurogenesis than CCI-operated mice. Thus, PPF1 demonstrated the ability to restore neurogenesis after chronic nerve injury. * *P<0.05 PPF1-treated CCI vs. vehicle-treated CCI, unpaired T-test.

[0201] Figure 28 shows data from histological examination of the hippocampus performed on wild-type mice described in Figure 24. The inflammatory marker, measured by the expression of CD68, was quantified. The results show that mice given vehicle treatment after CCI surgery had significantly more CD68-positive cells in the hippocampus than mice treated with PPF1 after CCI surgery. Animals treated with PPF1 have similar inflammation levels to the sham-operated group, indicating that PPF1 helps ameliorate neuroinflammation caused by chronic nerve injury. * *P<0.05 PPF1-treated CCI vs. vehicle-treated CCI, vehicle-treated sham-operated vs. vehicle-treated CCI, one-way ANOVA with Tukey post-hoc analysis.

[0202] Figure 29 shows data from the von Frey mechanical allodynia test performed on C57BL / 6J mice undergoing CCI surgery or sham surgery, using the timeline shown in Figure 24. 22-month-old mice were administered either PPF1 or vehicle control at 150 μL / day (intravenously via the tail vein) for 7 consecutive days in a continuous pulse administration regimen. Another group received gabapentin at 75 mg / kg (intraperitoneally) daily for 7 consecutive days. All treatments began 24 hours after CCI surgery or sham surgery. To determine the animals' tolerance to mechanical pressure, the subject's hind paw, weakened via the sciatic nerve, was stimulated with von Frey filaments of different thicknesses. The pressure at which the mice withdrew their hind paw was assessed and reported as the number of weeks after CCI surgery or sham surgery in Figure 29. This figure demonstrates that mice administered PPF1 after CCI surgery had significantly increased mechanical nociception at all time points assessed compared to mice treated with vehicle after CCI surgery. Conversely, gabapentin-treated mice only showed significant improvement in mechanical nociception at 2 weeks after CCI surgery, and were similar to vehicle-treated mice at other time points. Sham-operated mice showed highly significant responses to mechanical nociception at 3 and 5 weeks after surgery. Taken together, these data demonstrate that PPF1 improves peripheral pain for a longer period than standard treatment (gabapentin). *** , *****P<0.001, *P<0.0001 PPF1 vs. vehicle control, ANOVA with Tukey post hoc analysis * P<0.05 gabapentin vs. vehicle control, ANOVA with Tukey post hoc analysis; * , ** *P<0.05, *P<0.01 sham vs. vehicle control, ANOVA with Tukey post-hoc analysis.

[0203] Figure 30 shows data from a hot plate test performed on wild-type mice treated as shown in Figure 24, as described by Woolfe and Macdonald (Woolfe G. and Macdonald AD, J. Pharmacol. Exp. Ther. 80:300-07 (1944), which is incorporated by reference in its entirety). The hot plate was set to a temperature of 55°C. Mice were placed in the transparent cylinder for 30 minutes for acclimation. The cylinder was placed on the hot plate and a timer was started. The first observation of nocifensive behavior (e.g., hind paw licking, jumping) was recorded as the latency. If nocifensive behavior was observed, the animal was removed at a predetermined time interval, e.g., 30 seconds, to prevent tissue damage. Because repeated exposure to the test has been shown to alter sensitivity, mice were only tested at 2 and 5 weeks after CCI surgery. Figure 30 shows hot plate nocifensive latency 5 weeks after CCI surgery or sham surgery. PPF1 treatment was significantly less sensitive to hot plate stimulation than mice given CCI surgery and vehicle control, indicating a rescue effect by PPF1. ** P<0.01 sham surgery vs. CCI surgery; **** *P<0.0001 PPF1 vs. vehicle-treated CCI-operated mice, ANOVA with Tukey post-hoc analysis.

[0204] (2) Prevention of neuroinflammation in the spinal cord Another study similar to the one described above was conducted in 22-month-old C57BL / 6J mice. Cohorts of mice were treated with (1) PPF (PPF2) after CCI surgery, (2) vehicle after CCI surgery, (3) recombinant human albumin (rhAlb) after CCI surgery, or (4) vehicle after sham surgery. Mice were administered PPF2, recombinant human albumin, or vehicle control at 150 μL / day (intravenously via the tail vein) via a pulse dosing regimen for 7 consecutive days. All treatments were initiated 24 hours after CCI or sham surgery.

[0205] Figure 31 shows data from the hot plate test (described above) 35 days after CCI surgery treated according to the timeline in Figure 24. Mice given PPF2 treatment were significantly less sensitive to hot plate stimulation than mice given CCI surgery plus vehicle control. Mice treated with recombinant human albumin were significantly less sensitive to hot plate stimulation than mice given CCI surgery plus vehicle control, but not as sensitive as mice treated with PPF2. * P<0.05 rhAlb vs. vehicle-treated CCI-operated mice; *** *P<0.001 PPF2 vs. vehicle-treated CCI-operated mice, ANOVA with Tukey post-hoc analysis.

[0206] Figure 32 shows data from the von Frey mechanical allodynia test using these same mice at different time intervals, both before (baseline) and after CCI surgery. The pressure assessed when the mice withdrew their hind paw was expressed as the number of weeks after CCI or sham surgery. This figure demonstrates that mice administered PPF2 after CCI surgery had significantly increased mechanical nociception at all time points assessed compared with mice treated with vehicle or recombinant human albumin (rhAlb) after CCI surgery. This indicates that PPF (PPF2) improved pain for a longer period than either the control vehicle or albumin, the major protein component of PPF. Therefore, these effects appear to be mediated not through albumin but through other proteins present in PPF.* P<0.05, ** P<0.01, *** P<0.001, **** *P<0.0001 vs. vehicle control, ANOVA with Tukey post-hoc analysis.

[0207] Figure 33 shows the relative levels of myelin basic protein (MBP, detected by Abcam, ab40390 anti-rabbit antibody) in the distal sciatic nerve 5 weeks after the last dose of PPF (PPF1) in another similar experiment performed on 22-month-old mice as described above. * P<0.05, *** *P<0.001 vs. vehicle control, ANOVA with Tukey post-hoc analysis.

[0208] Figure 34 shows the relative levels of the S-100 Schwann cell marker in these mice. In both cases, the relative levels of these markers were elevated in PPF-treated mice undergoing CCI surgery compared to vehicle-control-treated mice undergoing CCI surgery. These results indicate that PPF promotes sciatic nerve repair mechanisms by increasing the expression of myelin and S-100 proteins. PPF also induces myelination repair mechanisms. ** P<0.01, *** *P<0.001 vs. vehicle control, ANOVA with Tukey post-hoc analysis.

[0209] FIG. 35 is a qualitative fluorescent microscopic representation of the data shown in FIGS.

[0210] Figures 36 and 37 show that BDNF and CD68, respectively, were detected in the dorsal horn of the spinal cord of treated mice 24 hours after CCI injury. Brain-derived neurotrophic factor (BDNF, detected by Abcam, anti-rabbit antibody ab108319) is secreted by activated microglia and has been shown to enhance spinal nociception (detection of painful stimuli) through synaptic facilitation and central sensitization-like mechanisms. Peripheral trauma-induced neuropathic pain is often accompanied by increased spinal expression of BDNF (Garraway SM, et al. Neural Plast. Article ID 9857201 (2016)). CD68 levels (detected by BioRad, anti-rat antibody MCA1957GA) were also measured. CD68 is a marker of activated microglia. Figures 36 and 37 show that PPF treatment 24 hours after CCI injury significantly reduces both BDNF and CD68 markers in the dorsal horn of the spinal cord, indicating inhibition of deleterious downstream events associated with the development of neuropathic pain and prevention of microglial activation. ** P<0.01, *** *P<0.001 vs. vehicle control, ANOVA with Tukey post-hoc analysis.

[0211] Figures 38 and 39 are fluorescence microscopy images of the data shown in Figures 39 and 37, respectively. The rectangle highlights the dorsal horn of the spinal cord analyzed at the L4-L6 lumbar segment. The images on the right side of the figures are higher magnification images of the rectangular areas on the left side of each figure.

[0212] b) Pain - Treatment 14 days after injury Figure 40 shows the protocol used for 22-month-old C57BL / 6J mice. Baseline von Frey hindpaw withdrawal thresholds for measuring mechanical allodynia were obtained 3–4 days before CCI surgery or sham surgery. Cohorts of mice were treated with: (1) PPF1 (PPF1) 14 days after CCI surgery, (2) vehicle 14 days after CCI surgery, (3) recombinant human albumin (rhAlb) 14 days after CCI surgery, or (4) vehicle 14 days after sham surgery. Mice were administered PPF1, recombinant human albumin, or vehicle control at 150 μL / day (intravenously in the tail vein) via a pulse administration regimen for 7 consecutive days. All treatments began 14 days after CCI surgery or sham surgery.

[0213] Figure 41 shows von Frey hindpaw withdrawal thresholds at baseline, 14, 21, 28, 35, 42, and 49 days after CCI surgery. Significant deficits were observed in all groups except the sham-operated group at day 14, indicating central sensitization in all CCI groups 2 weeks post-injury. This result persisted until 7 days (day 28) after PPF treatment was discontinued, indicating that PPF does not simply provide analgesia in this model. Instead, PPF treatment produces a mechanical effect that is not observed with vehicle or recombinant human albumin (rhAlb). This demonstrates that PPF significantly reduces pain that is well established before PPF treatment (which necessarily includes a central component) compared to vehicle controls. ** P<0.01, *** P<0.001, **** *P<0.0001 vs. vehicle control, ANOVA with Tukey post-hoc analysis.

[0214] Figures 42 and 43 show hot plate latency values ​​at 35 days (Figure 19) and 49 days (Figure 20) after CCI surgery. Both results demonstrate that PPF-treated mice have long-term reduced hot plate pain sensitivity, further supporting the observation that PPF acts through a mechanical effect rather than simply providing an analgesic effect. ** P<0.01, ANOVA with Tukey post hoc analysis.

[0215] Example 3 - Improvement of nerve myelination This example demonstrates that human plasma fractions such as PPF1, with enhanced safety and tolerability, can reverse age-related decline and neuroinflammation in the CNS. This example also demonstrates that plasma fraction-based therapeutic approaches can restore myelinogenic potential in models of aging and age-related disease, thereby addressing one or more age-related disease conditions, such as neuroinflammation and neurodegeneration.

[0216] In vivo mouse models were used to examine the impairment of myelination and the therapeutic utility of PPF1. Myelin basic protein (MBP) coverage was compared in both the hippocampus and cortex across various models, including aging, hyperhomocysteinemia (Hhcy), and cisplatin-induced cognitive impairment. Hippocampal PDGFRa-expressing oligodendrocyte progenitor cells (OPCs) were also quantified to recapitulate remyelination capacity. The effect of PPF1 on myelination was examined in aged mice, and increased myelin content was observed not only in the cortex but also in the hippocampus, which correlated with behavioral performance. Therefore, this example demonstrates that plasma fractions offer a multimodal therapeutic approach with the potential to restore myelin levels in models of aging and age-related disease, thereby ameliorating age-related behavioral decline.

[0217] histology Brains were harvested after saline perfusion, and fixed hemibrains were sectioned at 30 μm thickness. Free-floating sections were blocked with appropriate serum before incubation with primary antibodies at the following concentrations: MBP 1:1000, Abcam; OLIG2 1:1000, Invitrogen; PDGFRa 1:500, R&D Systems; CD68 1:1000, AbD Serotec; Iba-1 1:2500, Wako; DCX 1:2000, Millipore; BrdU 1:500 (antigen retrieval), Abcam. Y-maze—The assay chamber consisted of a three-arm maze with a start arm, a cued familiar arm, and a familiar arm. During a 5-minute training session, mice were allowed to explore the start and familiar arms of the maze. After a 4-hour delay, mice are tested with access to all three arms for 5 minutes, and duration and entries into each arm are recorded.

[0218] Image analysis MBP coverage and mean optical density were analyzed for the hippocampus, subregion CA1, and cortex using manual ROI and thresholding using Image Pro software (Media Cybernetics). OPC density was calculated by counting PDGFRα cells colocalized with Olig2 within the hippocampal ROI. Hyperhomocysteinemia (Hhcy) was induced in 12-week-old mice by feeding them Teklad Custom Diet TD.97345 (Envigo), which was deficient in folic acid, vitamins B6, and B12 and supplemented with methionine. For cisplatin studies, 7-month-old mice were administered 2.3 mg / kg cisplatin (232120, Calbiochem) via intraperitoneal (IP) injection.

[0219] Plasma fraction treatment reduced neuroinflammation and enhanced neurogenesis (Figures 44A-44D). Figure 44A shows a schematic diagram of the plasma fraction separation process. Figure 44B shows a schematic diagram of the study design in which 22- to 24-month-old wild-type male mice were administered PPF1 and analyzed 10 days (CD68 / IbA-1) or 6 weeks (BrdU / DCX). PPF1 treatment caused a reduction in microglia, as shown by quantification of CD68 and IbA-1 immunoreactivity in the hippocampus (Figure 44C). Figure 44D shows that PPF1 treatment improves cell survival and neurogenesis. All data shown are mean ± SEM. * p<0.05, ** p<0.01, *** p<0.001, Veh: vehicle.

[0220] Representative hippocampal images from 11- and 24-month-old mice show age-related myelin loss in the hippocampus (Figures 45A and 45B). The box on the right highlights the CA1 ROI shown in the image. Figure 45C shows that myelin coverage in the hippocampus and cortex did not change between 11- and 24-month-old mice. Figure 45D shows that the mean optical density of MBP signal was significantly increased in the hippocampus and CA1 in 11-month-old mice compared to 24-month-old mice. Figure 45E shows that PDGF-Ra in the hippocampus of 11- and 24-month-old mice. + Representative images of cells are shown. Figure 45F shows PDGFRa in the hippocampus. + Quantification of cell density is shown, and PDGFRa in the hippocampus + The data show that cell density did not change with age. Data shown are mean ± SEM, Mann-Whitney test. ** p<0.003, scale bars = 500 μm, 100 μm, 20 μm.

[0221] The Hhcy and cisplatin models do not show a deficit in myelin content. Specifically, Figure 46A shows the protocol for inducing HHcy in 12-week-old mice by feeding a folate-deficient diet for 10 weeks. Figure 46B shows that no differences were observed in myelin coverage, hippocampal MBP optical density, or OPC density measured by PDGFRa in the hippocampus. Figure 46C shows a schematic diagram of the protocol for inducing cognitive impairment in 7-month-old mice by IP administration of 2.3 mg / kg cisplatin. Figure 46D shows that no differences were observed in myelin coverage, hippocampal optical density, or OPC density measured by PDGFRa in the hippocampus. All data shown are mean ± SEM.

[0222] Aged mice treated with PPF1 show increased myelin content in the hippocampus and cortex (Figures 47A-47G). Figure 47A shows a schematic diagram of the experimental protocol in which 22-month-old mice were treated with PPF1 for 7 days and tissues were harvested 10 days later. Images of the hippocampal ROI (inset) and representative dentate gyrus show increased MBP expression in PPF1-treated mice (Figure 47B). PPF1 treatment increased myelin coverage and MBP optical density in the hippocampus and CA1 (Figure 47C). Figure 47D shows representative images of MBP expression in the cortex and ROI (outer dotted line). Figure 47E shows that increased MBP expression was observed in the cortex after PPF1 treatment. Figure 47F shows the relationship between PDGFRa and MBP expression in the cortex. + Figure 47G shows that no difference in OPC density was observed in the hippocampus. A significant correlation was observed between MBP expression and Y-maze performance (percent time in the novel arm) with PPF1 treatment. Spearman's correlation test, R = 0.7182. * p=0.0162, Mann-Whitney test, **** p<0.0001, *** p<0.0002, * p=0.03, scale bar=200 μm.

[0223] These data indicate that increasing myelin content provides a viable strategy for preventing age-related cognitive decline. Notably, administration of PPF1 improves various aspects of neuronal health in aged wild-type mice, including increasing myelin content.

[0224] In at least some of the above-described embodiments, one or more elements used in an embodiment may be used interchangeably in another embodiment unless such substitution is technically feasible. It will be apparent to those skilled in the art that various other omissions, additions, and modifications may be made to the methods and structures described above without departing from the scope of the subject matter described in the claims. All such modifications and alterations are intended to fall within the scope of the subject matter defined in the appended claims.

[0225] In general, those skilled in the art will understand that the terms used herein, particularly in the appended claims (e.g., the body of the appended claims), are intended as “open” terms throughout (e.g., the term “including” should be interpreted as “including but not limited to,” the term “having” should be interpreted as “having at least,” the term “includes” should be interpreted as “includes but is not limited to,” etc.). If a specific number recitation is intended in the appended claims, such intention will be expressly recited in the claim; in the absence of such recitation, those skilled in the art will further understand that no such intention exists. For example, as an aid to understanding, the appended claims may include the introductory phrases “at least one” and “one or more” to guide the recitation of the claims. However, the use of such phrases should not be construed as suggesting that introducing a claim recitation with the indefinite article "a" or "an" limits any particular claim that includes the claim recitation so introduced to embodiments that include only one such recitation, even if the same claim also includes the introductory phrase "one or more" or "at least one" and an indefinite article such as "a" or "an" (e.g., "a" and / or "an" should be construed to mean "at least one" or "one or more"). The same applies to the use of definite articles used to introduce claim recitations. Furthermore, even when a specific number is explicitly recited in an introduced claim recitation, those skilled in the art will understand that such a recitation should be construed to mean at least the recited number (e.g., the mere recitation of "two recitations" without any other modifier means at least two recitations, or more than two recitations).Furthermore, where conventional language similar to "at least one of A, B, and C, etc." is used, such syntax is typically intended in the sense that one of ordinary skill in the art would understand the conventional language (e.g., "a system having at least one of A, B, and C" includes, but is not limited to, systems having A only, B only, C only, A and B together, A and C together, B and C together, and / or A, B, and C together, etc.). Furthermore, where conventional language similar to "at least one of A, B, or C, etc." is used, such syntax is typically intended in the sense that one of ordinary skill in the art would understand the conventional language (e.g., "a system having at least one of A, B, or C" includes, but is not limited to, systems having A only, B only, C only, A and B together, A and C together, B and C together, and / or A, B, and C together, etc.). Those skilled in the art will further understand that virtually any disjunctive word and / or phrase presenting two or more alternative terms, wherever it appears in the specification, claims, or drawings, should be understood to contemplate the possibility of including one of the terms, either of the terms, or both terms. For example, the phrase "A or B" is understood to include the possibilities of "A" or "B" or "A and B."

[0226] Furthermore, when features or aspects of the present disclosure are described in terms of a Markush group, those skilled in the art will understand that the present disclosure is also being described in terms of any individual member or subgroup of members of the Markush group.

[0227] As will be understood by those skilled in the art, for all purposes, including for providing a written description, all ranges disclosed herein encompass all possible subranges and combinations of such subranges. Any ranges recited are fully descriptive and can be readily understood to be broken down into at least equal halves, thirds, quarters, fifths, tenths, etc. As a non-limiting example, each range discussed herein can be readily broken down into a lower third, a middle third, an upper third, etc. It will further be understood by one of ordinary skill in the art that all terms, such as "up to," "at least," "greater than," "less than," and the like, refer to ranges that include the recited numbers and that can subsequently be broken down into subranges as discussed above. Finally, it will be understood by one of ordinary skill in the art that a range includes each individual member. Thus, for example, a group having 1 to 3 substituents refers to the group having 1, 2, or 3 substituents. Similarly, a group having 1 to 5 substituents refers to the group having 1, 2, 3, 4, or 5 substituents, and so forth.

[0228] Although the foregoing invention has been described in some detail by way of example for clarity of understanding, it will be readily apparent to those skilled in the art in light of the teachings of this invention that certain changes and modifications can be made thereto without departing from the spirit or scope of the appended claims.

[0229] Accordingly, the foregoing merely illustrates the principles of the present invention. It will be appreciated that those skilled in the art will be able to devise various configurations that embody the principles of the present invention and are within its spirit and scope, even if not explicitly described or presented herein. Furthermore, all examples and conditional language recited herein are intended primarily to aid the reader in understanding the inventive principles and concepts that the inventors contribute to advancing the art, and should not be construed as being limited to such specifically recited examples and conditions. Furthermore, all statements herein reciting principles, aspects, and embodiments of the present invention, as well as specific examples thereof, are intended to encompass both structural and functional equivalents. Furthermore, such equivalents are intended to include both currently known equivalents and future-developed equivalents, i.e., any elements developed to perform the same function, regardless of structure. Furthermore, nothing disclosed herein is intended to be open to the public, regardless of whether such disclosure is explicitly recited in the claims.

[0230] The scope of the invention is not intended to be limited to the exemplary embodiments shown and described herein. Rather, the scope and spirit of the present invention is embodied by the appended claims. In the claims, 35 U.S.C. 112(f) or 35 U.S.C. 112(6) is expressly defined to be invoked with respect to a limitation in a claim only if the precise phrase "means for" or the precise phrase "step for" is recited at the beginning of such limitation in the claim; if such precise phrases are not used in the limitation in the claim, 35 U.S.C. 112(f) or 35 U.S.C. 112(6) is not invoked.

[0231] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority by the filing date of U.S. Application No. 17 / 985,721, filed November 11, 2022, the disclosure of which is incorporated herein by reference.

Claims

1. 1. A method of restoring myelin levels and / or improving nerve conductance, comprising: A method comprising administering an effective amount of a plasma fraction to a subject diagnosed with a condition associated with myelin degeneration.

2. 2. The method of claim 1, wherein the plasma fraction is a plasma protein fraction (PPF).

3. 3. The method of claim 2, wherein the plasma protein fraction is a commercially available plasma protein fraction.

4. 3. The method of claim 2, wherein the plasma protein fraction has a total protein content of at least 83 percent and less than 95 percent albumin and no more than 17 percent globulins.

5. 5. The method of claim 4, wherein the plasma protein fraction contains 1% or less of gamma globulin.

6. 10. The method of claim 1, comprising restoring myelin levels in the subject.

7. 10. The method of claim 1, comprising improving neural conductance in the subject.

8. 2. The method of claim 1, wherein the condition associated with myelin degeneration is an age-related neurodegenerative and / or neuroinflammatory disease.

9. 10. The method of claim 1, wherein the condition associated with myelin degeneration is myelopathy associated with post-surgical recovery.

10. 2. The method of claim 1, wherein the plasma fraction is derived from plasma obtained from a pool of young individuals.

11. 10. The method of claim 1, wherein the plasma fraction is produced from mammalian blood products.

12. 12. The method of claim 11, wherein the mammalian blood product is a human blood product.

13. The method of claim 1 , wherein the subject is a mammal.

14. 14. The method of claim 13, wherein the mammal is a human.

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