Dosage regimens for the treatment of cognitive and movement disorders with blood plasma and blood plasma derivatives - Patents.com

Abstract

Methods and compositions for treating and / or preventing age-related conditions are described. The compositions used in the methods include blood plasma and blood plasma fractions derived from blood plasma that are effective in treating and / or preventing age-related conditions, such as cognitive impairment. The methods involve pulsed dosing regimens of blood plasma or blood plasma fractions.

Description

[Technical field]

[0001] The present invention relates to the prevention and treatment of age-related diseases. The present invention relates to the use of blood products, such as blood plasma fractions, for treating and / or preventing age-related conditions, such as cognitive impairment, movement disorders and neuroinflammation, using various dosing regimens. [Background technology]

[0002] The following is provided as background information only 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, blindness, 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 most important contributing factor to the neurological and cognitive impairments associated with these conditions. As such, aging remains the single most prevalent risk factor for dementia-related neurodegenerative diseases such as Alzheimer's disease (AD) (Bishop, NA et al., Neural mechanisms of ageing and cognitive decline. Nature, 464(7288), 529-535(2010); Heeden, T. et al., Insights into the ageing mind: a view from cognitive neuroscience. Nat. Rev. Neurosci. 5(2), 87-96(2004); Mattson, MP, et al., Ageing 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 neurodegeneration and decline in functions such as cognitive or motor skills can seriously affect quality of life. Treatments for cognitive decline, motor disorders, and neurodegenerative diseases have had some success in preventing and reversing disorders. Therefore, it is important to identify new treatments to maintain cognitive integrity by protecting against, countering, or reversing the effects of aging. Furthermore, as new treatments are developed, dosing norms must be considered to optimize the effectiveness of these treatments.

[0005] Parabiotic experiments between old and young mice have shown that cognitive function can be improved in old mice by heterochronic blood exchange with young mice, but a recent report found that a single exchange of young blood did not enhance neurogenesis in old mice (Rebo, J. et al. A single heterochronic blood exchange reveals rapid inhibition of multiple tissues by old blood. Nat. Comm. (2016)). Furthermore, it is suspected that cognitive function and neurogenesis resulting from the injection of young plasma are linked. Therefore, a dosing paradigm using blood plasma or blood plasma fractions to stimulate neurogenesis and improve cognitive function has not yet been described. Summary of the Invention

[0006] The present invention is based on the production and use of blood products for treating and / or preventing age-related diseases such as, for example, cognitive impairment conditions, age-related dementia, motor function disorders, neuroinflammation and neurodegenerative diseases. The present invention recognizes, inter alia, the need for new therapeutic approaches for the treatment and / or prevention of cognitive impairment, age-related dementia, motor disorders, neuroinflammation and neurodegenerative diseases. Derived from blood and blood plasma, the present compositions of the present invention relate to a solution for the failures and shortcomings of current therapeutic approaches via the use of blood plasma fractions that show efficacy in the treatment and / or prevention of cognitive impairment, age-related dementia, motor disorders, neuroinflammation and neurodegenerative diseases.

[0007] The present invention recognizes that blood plasma proteins have an average half-life of 2-3 days. The present invention uses a blood plasma or plasma fraction dosing regimen that optimizes neurogenesis, cell survival, reduced neuroinflammation, and improved cognitive or motor function in treated subjects. The dosing regimen of the present invention has been found to induce all of these processes in subjects (neurogenesis, cell survival, improved cognition, reduced neuroinflammation, improved motor function), all of which are found to be active even weeks after the last dose.

[0008] An embodiment of the invention includes treating a subject diagnosed with a cognitive disorder by administering an effective amount of blood plasma or a plasma fraction to the subject. Another embodiment of the invention includes administering an effective amount of blood plasma or a plasma fraction and then monitoring the subject for improvement in cognitive function. Another embodiment of the invention includes treating a subject diagnosed with a cognitive disorder by administering an effective amount of blood plasma or a plasma fraction to the subject, where the blood plasma or plasma fraction is administered in a manner that results in improved cognitive function or neurogenesis.

[0009] An embodiment of the invention includes treating a subject diagnosed with a neurodegenerative movement disorder, such as, for example and not by way of limitation, Parkinson's disease, by administering to the subject an effective amount of blood plasma or a plasma fraction. Another embodiment of the invention includes administering an effective amount of blood plasma or a plasma fraction and subsequently monitoring the subject for improvement in motor function. Another embodiment of the invention includes treating a subject diagnosed with a neurodegenerative movement disorder by administering to the subject an effective amount of blood plasma or a plasma fraction, where the blood plasma or plasma fraction is administered in a manner that results in improvement in motor function or neurogenesis.

[0010] An embodiment of the invention includes treating a subject diagnosed with neuroinflammation or a neuroinflammation-related disorder by administering an effective amount of blood plasma or a plasma fraction to the subject. Another embodiment of the invention includes administering an effective amount of blood plasma or a plasma fraction and then monitoring the subject for a reduction in neuroinflammation. Another embodiment of the invention includes treating a subject diagnosed with neuroinflammation or a neuroinflammation-related disorder by administering an effective amount of blood plasma or a plasma fraction to the subject, wherein the blood plasma or plasma fraction is administered in a manner that results in a reduction in neuroinflammation.

[0011] Another embodiment of the invention includes administering blood plasma or plasma fractions via a dosing regimen of at least 2 consecutive days. Further embodiments of the invention include administering blood plasma or plasma fractions via a dosing regimen of at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 consecutive days (referred to herein as "pulsed dosing", "pulsed administration", "pulse dosing", "pulse administration" or "pulsed administered"). Yet another embodiment of the invention includes administering blood plasma or plasma fractions via a dosing regimen of at least 2 consecutive days and after the date of the last administration. Another embodiment of the invention includes administering blood plasma or plasma fractions via a dosing regimen of 2 to 14 non-consecutive days, where each interval between administrations may be 0 to 3 days, respectively. Another embodiment of the invention includes monitoring the subject for improved cognitive or motor function, reduced neuroinflammation, or improved neurogenesis at least 3 days after the date of the last administration. Another embodiment of the invention includes monitoring the subject for improved cognitive or motor function, reduced neuroinflammation, or improved neurogenesis past the average half-life of the protein in blood plasma or plasma fractions.

[0012] In some cases, pulsatile dosing according to the invention includes administration of a first set of doses, e.g., as described above, followed by a dosing-free period, e.g., a "drug-free period", followed by administration of another dose or set of doses. The duration of this "drug-free period" may vary, but in some embodiments is 7 days or more, e.g., 10 days or more, including 14 days or more, and in some cases the dosing-free period ranges from 15 to 365 days, e.g., 30-90 days and 30-60 days. As such, method embodiments include non-chronic (i.e., non-continuous) dosing, e.g., non-chronic administration of blood plasma products. In some embodiments, the pattern of pulsatile dosing followed by a drug-free period is repeated as many times as desired, where in some cases the pattern is continued for one year or more, e.g., two years or more, up to and including the life of the subject. Another embodiment of the invention involves administering blood plasma or plasma fractions via a dosing regimen of 5 consecutive days with 2-3 dosing-free days followed by 2-14 consecutive days of dosing.

[0013] The present invention also recognizes that differences in protein content between various blood plasma fractions (e.g., fractions, eluates, plasma protein fractions, human albumin solutions) may be involved in preventing and / or ameliorating certain cognitive or motor disorders and alleviating neurodegenerative diseases. By way of example and not limitation, embodiments of the present invention demonstrate that the mere high albumin concentration of recombinant human albumin or human albumin solution (HAS) formulations is not the driving force behind the improved cognition, improved motor function, reduced neuroinflammation, cell survival, or neurogenesis associated with plasma protein fraction (PPF) formulations with lower albumin concentrations.

[0014] Blood and blood plasma from young donors have been shown to improve and reverse the existing effects of brain aging, including at the 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 in part to blood plasma fractions and eluates that have traditionally been used to treat shock in patients, and to the discovery that they are effective as a method of treatment for age-related cognitive impairment, motor function decline, and neuroinflammation or neurodegeneration-related diseases.

[0015] According to aspects of the invention, methods of treating age-associated cognitive impairment, age-related dementia, movement disorders, neuroinflammation and / or neurodegenerative diseases using blood product fractions of blood plasma are then provided. Method aspects include administering blood plasma fractions to individuals suffering from or at risk of developing age-associated cognitive impairment, movement disorders, neuroinflammation or neurodegenerative diseases. Additional method aspects include administering blood plasma fractions derived from a pool of donors of a particular age range to individuals suffering from or at risk of developing age-associated cognitive impairment, movement disorders, neuroinflammation or neurodegenerative diseases. Further method aspects include administering blood plasma or plasma fractions using a pulsed dosing regimen. Also provided are reagents, equipment and kits thereof used to practice the subject methods.

[0016] In embodiments, the blood plasma fraction may be one of several blood plasma fractions obtained from a blood fractionation method, such as, for example, the Cohn fractionation method described below. In another embodiment, the blood plasma fraction may be of the type referred to herein as a "plasma fraction", which is a solution composed of normal human albumin, alpha and beta globulins, gamma globulins, and other proteins, either individually or in complexes. In another embodiment, the blood plasma fraction may be of the type of blood plasma fraction known to those skilled in the art as a "plasma protein fraction" (PPF). In another embodiment, the blood plasma fraction may be a "human albumin solution" (HAS) fraction. In yet another embodiment, the blood plasma fraction may be one in which substantially all of the clotting factors have been removed to preserve the efficacy of the fraction with reduced risk of thrombosis. An embodiment of the invention may also include administering a fraction derived from, for example, a young donor or a pool of young donors. Another embodiment of the invention may include monitoring subjects treated with the blood plasma fraction for improved cognition, improved motor function, reduced neuroinflammation, or increased neurogenesis.

[0017] An embodiment of the present invention includes treating a subject diagnosed with cognitive impairment, neurodegenerative movement disorder, or neuroinflammation-related disease by administering an effective amount of blood plasma or plasma fraction to the subject. Another embodiment of the present invention includes administering an effective amount of blood plasma or plasma fraction and then monitoring the subject for improved cognitive function, improved motor function, reduced neuroinflammation, or increased neurogenesis. Another embodiment of the present invention includes administering an effective amount of blood plasma or plasma fraction via a dosing regimen of at least two consecutive days and monitoring the subject for improved cognitive function, improved motor function, reduced neuroinflammation, or increased neurogenesis at least two days after the date of the last administration. Further embodiments of the present invention include administering an effective amount of blood plasma or plasma fraction via a dosing regimen of at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 days and monitoring the subject for improved cognitive function, improved motor function, reduced neuroinflammation, or increased neurogenesis at least three days after the date of the last administration. Yet another embodiment of the invention includes administering an effective amount of blood plasma or a plasma fraction via a dosing regimen of at least two consecutive days and monitoring for improved cognition, improved motor function, reduced neuroinflammation or increased neurogenesis after the average half-life of the protein in the blood plasma or plasma fraction is reached after the date of the last administration.

[0018] An embodiment of the invention includes treating a subject diagnosed with cognitive impairment, motor dysfunction, neuroinflammation, or reduced neurogenesis by administering an effective amount of blood plasma or plasma fraction to the subject, and the subject is subjected to an exercise regimen after administration. Another embodiment of the invention includes subjecting the subject to an exercise regimen prescribed for the subject. Another embodiment of the invention includes subject exercising at a higher intensity and / or with a higher frequency than the subject who exercised prior to administration. Another embodiment of the invention includes subject exercising at a similar intensity and / or frequency to the subject who exercised prior to administration.

[0019] An embodiment of the present invention includes treating a subject diagnosed with cognitive impairment, motor dysfunction, neuroinflammation, or reduced neurogenesis by administering an effective amount of blood plasma or plasma fraction to the subject, the subject undergoing, to be undergoing, or having undergone stem cell therapy. Another embodiment of the present invention includes administering an effective amount of blood plasma or plasma fraction to the subject, the subject undergoing, to be undergoing, or having undergone stem cell therapy, the stem cells used in the therapy can be embryonic stem cells, non-embryonic stem cells, induced pluripotent stem cells (iPSCs), umbilical cord blood stem cells, amniotic fluid stem cells, etc. Another embodiment of the present invention includes treating a subject diagnosed with traumatic spinal cord injury, stroke, retinal disease, Huntington's disease, Parkinson's disease, Alzheimer's disease, hearing loss, heart disease, rheumatoid arthritis, or severe burns, the subject undergoing, to be undergoing, or having undergone stem cell therapy, with an effective amount of blood plasma or plasma fraction.

[0020] 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 description of the drawings]

[0021] [Figure 1] A shows the distance traveled in the open field test in mice treated with PPF1 using a pulse dosing and 3x / weekly dosing regimen, and B shows the time spent in the center in the open field test in mice treated with PPF1 using a pulse dosing and 3x / weekly dosing regimen. [Diagram 2] FIG. 1 shows body weight over time for mice treated with PPF1 using pulse dosing and 3× / weekly dosing regimens. [Diagram 3] FIG. 13 shows the number of DCX-labeled cells within the granular layer of the dentate gyrus in mice treated with PPF1 using a pulse or 3× / week dosing regimen. [Figure 4]FIG. 1 shows the number of BrdU-labeled cells within the granular layer of the dentate gyrus in mice treated with PPF1 using a pulse or 3× / week dosing regimen. [Diagram 5] FIG. 1 shows the number of DCX-labeled cells in the granular layer of the dentate gyrus in mice treated with PPF1, young human plasma ("YP") or old human plasma ("OP") using a pulse or 3x / week dosing regimen. [Figure 6] FIG. 1 shows the number of BrdU-labeled cells in the granular layer of the dentate gyrus in mice treated with PPF1, YP or OP using a pulse or 3× / week dosing regimen. [Figure 7] FIG. 1 shows the latency to find the target hole per trial per day for mice pulsed with PPF1 or YP. [Figure 8] FIG. 1 shows the number of DCX-labeled cells within the granular layer of the dentate gyrus in groups of mice treated with either young human plasma (YP), old human plasma (OP) or PPF1 using a pulsed regimen. [Figure 9] FIG. 1 shows the number of BrdU-labeled cells within the granular layer of the dentate gyrus in groups of mice treated with either young human plasma (YP), old human plasma (OP) or PPF1 using a pulsed regimen. [Figure 10] Figure 1 shows the percentage of total entries into familiar or novel lanes out of total entries into each lane by treatment group in the Y-maze test.12-month-old mice were pulse-treated with PPF1 or 5x concentrated PPF1. [Figure 11] Figure 1 shows the ratio of entries into a novel lane to a familiar lane in a Y-maze test.12-month-old mice were pulse-treated with PPF1 or 5x concentrated PPF1. [Figure 12] FIG. 1 shows the number of BrdU-labeled cells per hippocampal slice in 12-month-old mice pulsed with PPF1 or 5× concentrated PPF1. [Figure 13]FIG. 1 shows the number of DCX-labeled cells per hippocampal slice in 12-month-old mice pulsed with PPF1 or 5× concentrated PPF1. [Figure 14] FIG. 1 shows the number of DCX-labeled cells in the granular layer of the dentate gyrus in 10.5-month-old NSG mice pulse-dosed with PPF1 or saline using one of the following dosing regimens: (1) for 5 consecutive days [PPF1-5d]; (2) for 7 consecutive days [PPF1-7d]; (3) for 5 consecutive days with an additional 5 consecutive days of dosing occurring 6 weeks after completion of the first dosing ("booster") [PPF1-5d-B]; or (4) for 7 consecutive days with an additional 7 consecutive days of dosing occurring 6 weeks after completion of the first dosing ("booster") [PPF1-7d-B]. [Figure 15] FIG. 1 shows the number of BrdU-labeled cells in the granular layer of the dentate gyrus in 10.5-month-old NSG mice pulsed with PPF1 or saline using one of the following dosing regimens: (1) for 5 consecutive days [PPF1-5d]; (2) for 7 consecutive days [PPF1-7d]; (3) for 5 consecutive days with an additional 5 consecutive days of dosing occurring 6 weeks after completion of the first dosing ("booster") [PPF1-5d-B]; or (4) for 7 consecutive days with an additional 7 consecutive days of dosing occurring 6 weeks after completion of the first dosing ("booster") [PPF1-7d-B]. [Figure 16] FIG. 1 shows the number of EdU-labeled cells in the granular layer of the dentate gyrus in 10.5-month-old NSG mice pulsed with PPF1 or saline using one of the following dosing regimens: (1) for 5 consecutive days [PPF1-5d]; (2) for 7 consecutive days [PPF1-7d]; (3) for 5 consecutive days with an additional 5 consecutive days of dosing occurring 6 weeks after completion of the first dosing ("booster") [PPF1-5d-B]; or (4) for 7 consecutive days with an additional 7 consecutive days of dosing occurring 6 weeks after completion of the first dosing ("booster") [PPF1-7d-B]. [Figure 17] FIG. 13 shows the number of DCX-labeled cells in the granular layer of the dentate gyrus in 3- or 6-month-old NSG animals treated with PPF1 or saline in the presence or absence of a running wheel. [Figure 18]FIG. 13 shows the number of Ki67 positively labeled cells in the granular layer of the dentate gyrus in 3- or 6-month-old NSG animals treated with PPF1 or saline in the presence or absence of a running wheel. [Figure 19] FIG. 13 shows the number of BrdU positively labeled cells in the granular layer of the dentate gyrus in 3- or 6-month-old NSG animals treated with PPF1 or saline in the presence or absence of a running wheel. [Figure 20] FIG. 1 shows the number of wheel revolutions at given times in 11-month-old NSG mice pulsed with either PPF1 or saline control, where the shaded area indicates the dark period and the boxed area indicates when the hot plate test was administered. [Figure 21] A, Number of BrdU labeled cells in the granular layer of the dentate gyrus in three treatment groups of 10.5 month old NSG mice treated with young plasma, recombinant human albumin ("rhalbumin") and saline control. B, Number of DCX labeled cells in the granular layer of the dentate gyrus in three treatment groups of 10.5 month old NSG mice treated with young plasma, recombinant human albumin ("rhalbumin") and saline control. [Figure 22] FIG. 1 shows the extent of increased neuronal network activity in dissociated mixed neurons from mouse E16 cortex treated with control, PPF1, HAS1 or rhAlbumin. [Figure 23] FIG. 1 shows four regimens of administration of 8 week old (young) NSG mice receiving clarified aged human plasma (aged plasma) or saline. [Figure 24]A shows VCAM-1 positive labeling in the hippocampus of 8 week old (young) NSG mice treated with twice weekly dosing of old plasma 48 hours after the last dose. B shows VCAM-1 positive labeling in the hippocampus of 8 week old (young) NSG mice treated with three times weekly dosing of old plasma 48 hours after the last dose. C shows VCAM-1 positive labeling in the hippocampus of 8 week old (young) NSG mice treated with pulsed dosing of old plasma 48 hours after the last dose. D shows VCAM-1 positive labeling in the hippocampus of 8 week old (young) NSG mice treated with pulsed dosing of old plasma 21 days after the last dose. [Diagram 25] A, number of DCX positive cells in the dentate gyrus of 8 week old (young) NSG mice treated with twice weekly doses of aged plasma 48 hours after the last dose. B, number of DCX positive cells in the dentate gyrus of 8 week old (young) NSG mice treated with three weekly doses of aged plasma 48 hours after the last dose. C, number of DCX positive cells in the dentate gyrus of 8 week old (young) NSG mice treated with pulsed dosing of aged plasma 21 days after the last dose. [Figure 26] Barnes maze escape time course for 8 week old (young) NSG mice treated with aged plasma and saline, showing the time to reach and enter the escape hole. Mice were treated with aged human plasma or saline for 7 consecutive days and examined 4 weeks after the last injection. [Figure 27] Figure 1 shows the mean escape latency in the last three Barnes maze trials on day 4 of testing of 8 week old (young) NSG mice treated with aged human plasma or saline for 7 consecutive days. Testing was performed 4 weeks after injection. [Figure 28] Figure 1 shows the difference in escape latency between Barnes maze trials 1 and 3 in 8 week old (young) NSG mice treated with aged human plasma or saline for 7 consecutive days. Testing occurred 4 weeks after the last injection. [Figure 29]FIG. 1 shows quantitative polymerase chain reaction (qPCR) results quantifying mRNA levels of DCX, vesicular glutamate receptor (vglut1), synapsin 1 (syn1), beta III tubulin (tuj1) and brain-derived neurotrophic factor (bdnf) in 8-week-old (young) NSG mice treated with aged human plasma or saline for 7 consecutive days. [Diagram 30] FIG. 13 shows the dosing regimen for 8 week old (young) NSG mice treated with 35 mg / kg kainic acid or saline, followed by daily treatment with PPF1 or saline for 5 consecutive days. [Diagram 31] A shows the percentage of CD68-positive area in the CA1 region of the hippocampus of the routinely treated mice shown in Figure 28. B shows the percentage of GFAP-positive area in the CA1 region of the hippocampus of the routinely treated mice shown in Figure 28. [Diagram 32] Figure 1 shows the number of cells stained for BrdU in the dentate gyrus of 6-month-old NSG mice pulsed for 7 consecutive days with PPF1 or saline control along with concomitant administration of BrdU. The first two columns constitute the cohort analyzed 7 days after the last treatment of PPF1 / saline control and BrdU, the second two constitute the cohort analyzed 14 days after the last treatment of PPF1 / saline control and BrdU. [Diagram 33] FIG. 1 shows an increase in proliferating cells (Ki67+) in the dentate gyrus of 6-month-old NSG mice 10 days after completion of a pulse regimen with PPF1. [Diagram 34] FIG. 1 shows sections of the dentate gyrus and subventricular zone in 6-month-old NSG mice 10 days after completion of a pulse dosing regimen with PPF1. [Diagram 35]A, Cell fate of cells in the dentate gyrus of 6-month-old NSG mice treated with either PPF1 or saline control by a 7-day pulse regimen. BrdU was administered for 5 consecutive days immediately prior to the start of the pulse regimen. NeuN+ colocalization with BrdU indicates the extent to which neural progenitor cells become neurons. The extent of GFAP+ colocalization with BrdU indicates the extent to which neural progenitor cells become astrocytes. B, Results from an experiment similar to FIG. 35A, but NSG mice were 12 months old. [Diagram 36] A shows the cell fate of cells in the dentate gyrus of 3-month-old NSG mice treated with either aged plasma or saline control by a 7-day pulse regimen. BrdU was administered for 5 consecutive days immediately prior to the start of the pulse regimen. NeuN+ colocalization with BrdU indicates the extent to which neural progenitor cells become neurons. B shows results from the experiment detailed in FIG. 36A, where the extent of GFAP+ colocalization with BrdU indicates the extent to which neural progenitor cells become astrocytes. [Figure 37] FIG. 1 shows the number of cFos positive cells in (A) the whole brain, (B) the cortex, and (C) the isocortex of 18-month-old mice treated with a 7-day pulse regimen of PPF1 or saline. [Figure 38] FIG. 1 shows the number of cFos positive cells in (A) the frontal cortex, (B) the orbital cortex, (C) the intralimbic cortex, and (D) the prelimbic cortex of 18-month-old mice treated with a 7-day pulse dosing regimen of PPF1 or saline. [Figure 39] FIG. 1 shows the number of cFos positive cells in (A) the accessory olfactory nucleus and (B) the olfactory tubercle of 18-month-old mice treated with a 7-day pulse regimen of PPF1 or saline. [Diagram 40] FIG. 1 shows voxel statistics-based visualization of regional cortical activation in the frontal cortex (FRP), orbital cortex (ORB), intralimbic cortex (ILA), and prelimbic cortex (PL) and accessory olfactory nucleus (AON) of 18-month-old mice treated with a 7-day pulse dosing regimen of PPF1 or saline. [Diagram 41]A shows the percentage of CD68 immunoreactive area in the hippocampus of 22-month-old C57BL / 6J wild-type mice treated with a 7-day pulse regimen of PPF1 or saline control. B shows the percentage of Iba-1 immunoreactive area in the hippocampus of 22-month-old C57BL / 6J wild-type mice treated with a 7-day pulse regimen of PPF1 or saline control. C shows the percentage of GFAP immunoreactive area in the hippocampus of 22-month-old C57BL / 6J wild-type mice treated with a 7-day pulse regimen of PPF1 or saline control. [Diagram 42] A shows the percent change in BrdU staining in PPF1-treated, 23-month-old wild-type C57BL / 6J mice compared to saline controls after 6, 9, and 12 weeks of dosing with a 7-day pulsed dosing regimen for consecutive days, and B shows the percent change in DCX staining in PPF1-treated, 23-month-old wild-type C57BL / 6J mice compared to saline controls after 6, 9, and 12 weeks of dosing with a 7-day pulsed dosing regimen for consecutive days. [Diagram 43] A and B show the results of body weight measurements of 4-4.5 month old male alpha-synuclein mice (line 61) (a model of Parkinson's disease) treated with a pulsed dosing regimen of PPF1 or vehicle control for 7 consecutive days. [Diagram 44] Figure 14. Nesting results in 4-4.5 month old male alpha-synuclein mice (line 61) (model for Parkinson's disease) treated with a pulsed dosing regimen for 7 consecutive days with PPF1 or vehicle control. [Diagram 45] A and B show the results of improvement of pasta chewing and associated movement, respectively, in 4-4.5 month old male alpha-synuclein mice (line 61) (a model of Parkinson's disease) treated with a pulsed dosing regimen for 7 consecutive days with PPF1 or vehicle control. [Diagram 46]FIG. 1 shows wire suspension test results in 4-4.5 month old male alpha-synuclein mice (line 61) (model of Parkinson's disease) treated with a pulsed dosing regimen for 7 consecutive days with PPF1 or vehicle control. [Figure 47] A shows the different beam shapes and sizes used in five different beam walking tests of increasing difficulty. B shows the results of five different beam walking tests in 4-4.5 month old male alpha-synuclein mice (line 61) (a model of Parkinson's disease) treated with a pulsatile dosing regimen for 7 consecutive days with PPF1 or vehicle control. Beam walking tests were performed 72 hours after the last treatment dose. C shows the results of five different beam walking tests in 4-4.5 month old male alpha-synuclein mice (line 61) (a model of Parkinson's disease) treated with a pulsatile dosing regimen for 7 consecutive days with PPF1 or vehicle control. Beam walking tests were performed 3 weeks after the last treatment dose. [Figure 48] A-F show histological results of striatal and hippocampal staining in 4-4.5 month old male alpha-synuclein mice (line 61) (a model of Parkinson's disease) treated with a continuous 7 day pulsed dosing regimen with PPF1 or vehicle control. Histological markers examined included CD68, Iba-1 and NeuN. [Figure 49] FIG. 1 shows Barnes maze escape latency in 12-month-old NSG mice treated with a pulsed dosing regimen for 7 consecutive days with PPF1, HAS1 or vehicle control. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0022] 1. Introduction The present invention relates to the identification and discovery of methods and compositions for the treatment and / or prevention of cognitive and motor disorders, including age-related dementia or decline in motor function, and / or neurodegenerative diseases. Described herein are methods and compositions for the treatment of subjects suffering from such disorders, which are aspects of the present invention. Described herein are also dosing regimens that result in reduced neurogenesis or neuroinflammation and / or improved cognition or movement in subjects suffering from cognitive or motor disorders. The methods and compositions described herein are useful in preventing cognitive and motor disorders, age-related dementia, neuroinflammation, and / or neurodegenerative diseases, improving symptoms of cognitive and motor disorders, age-related dementia, neuroinflammation, and / or neurodegenerative diseases, slowing down the progression of age-related cognitive or motor disorders, age-related dementia, neuroinflammation, and / or neurodegenerative diseases, and / or reversing the progression of age-related cognitive and motor disorders, age-related dementia, neuroinflammation, and / or neurodegenerative diseases. The practice of the invention includes the use of blood plasma fractions as a treatment, for example, one or more fractions or eluates obtained from a blood fractionation process, such as the Cohn fractionation process described below. An embodiment of the invention includes the use of a plasma fraction (hereinafter a solution composed of normal human albumin, alpha and beta globulins, gamma globulins, and other proteins, either individually or in combination, referred to as a "plasma fraction"). Another embodiment of the invention includes the use of a plasma protein fraction (PPF) as a treatment. Another embodiment of the invention includes the use of a human albumin solution (HAS) fraction as a treatment. Yet another embodiment includes the use of an eluate from a blood fractionation process, such as eluate I or eluate II / III, described below. Additional embodiments include blood plasma fractions that have been stripped of substantially all clotting factors to retain efficacy while reducing the risk of thrombosis (see, for example, U.S. Patent Application Publication Nos. 62 / 236,710 and 63 / 376,529, which are incorporated herein by reference in their entirety).

[0023] Before describing the invention in detail, it is to be understood that the invention is not limited to the particular methods or compositions described, as such may, of course, vary. It is also 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 invention will be limited only by the appended claims.

[0024] The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein is to 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.

[0025] Where a range of values ​​is provided, each intervening value between the lower and upper endpoints of the range, to one tenth of the unit of the lower endpoint, is also specifically disclosed unless the context clearly dictates otherwise. Each smaller range between a stated or intervening value in a stated range and any other stated or intervening value in that stated range is encompassed within the invention. Depending on the specifically excluded endpoints in the stated range, the upper and lower endpoints of these smaller ranges may be independently included or excluded in the range, and each range in which one, neither, or both endpoints are included in the smaller range is also encompassed within the invention. When a stated range includes one or both endpoints, ranges excluding either or both of those included endpoints are also included in the invention.

[0026] It is noted that the claims may be drafted to exclude any element. As such, this statement is intended to serve as a predicate for the use of exclusive terminology such as "solely," "only," and the like in conjunction with recitation of claim elements or the use of a "negative" limitation.

[0027] As will be apparent to one of skill in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has distinct components and features which may be readily separated from or combined with the features of any of the other several embodiments without departing from the scope or spirit of the invention. Recited methods can be carried out in the order of events recited or in any other order which is logically possible.

[0028] 2.Definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of this invention, some possible and preferred methods and materials are described herein. All publications mentioned herein are incorporated by reference to disclose and describe the methods and / or materials with which the publications are cited. It is understood that the present disclosure supersedes the disclosure of the incorporated publication to the extent that there is a conflict.

[0029] 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 plurality of such cells, and reference to "the peptide" includes one or more peptides and equivalents thereof known to those skilled in the art, e.g., polypeptides.

[0030] In describing the methods of the invention, the terms "host", "subject", "individual" and "patient" are used interchangeably and refer to a mammal in need of such treatment according to the disclosed methods. 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 farm animal. In some embodiments, the subject is a pet. In some embodiments, the subject is a mammal. In certain embodiments, the subject is a human. Other subjects can include domestic pets (e.g., dogs and cats), farm animals (e.g., cows, pigs, goats, horses, etc.), rodents (e.g., as animal models of disease, e.g., mice, guinea pigs, and rats), and non-human primates (e.g., chimpanzees and monkeys). As such, subjects of the invention include, but are not limited to, mammals, e.g., humans and other primates, e.g., chimpanzees and other ape and monkey species, and in certain embodiments, the subject is a human. The term subject is also intended to include a human or organism of any age, weight or other physical characteristic, where the subject may be an adult, a child, an infant or a neonate.

[0031] By "young" or "young individual" is meant an individual who is a chronological age of 30 years or less, e.g., 40 years or less, including 25 years or less or 22 years or less, e.g., 35 years or less. In some cases, the individual who serves as the source of the blood product, including young plasma, is one who is 10 years or less, e.g., 5 years or less, including 1 year or less. In some cases, the subject is a newborn and the source of the plasma product is umbilical cord blood, where the plasma product is collected from the newborn's umbilical cord blood. As such, "young" and "young individual" may refer to a subject who is between 0 and 40 years old, e.g., 0, 1, 5, 10, 15, 20, 25, 30, 35, or 40 years old. In other examples, "young" and "young individual" may refer to a biological age (as opposed to a chronological age), such as an individual who does not exhibit levels of inflammatory cytokines in plasma exhibited by relatively older individuals. Conversely, "young" and "young individual" may refer to biological age (as opposed to chronological age), such as an individual who exhibits elevated levels of anti-inflammatory cytokines in plasma compared to 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 individuals and older individuals is at least 1.5-fold. Similarly, the fold difference between older and younger individuals in other inflammatory cytokines 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, e.g., the individual does not have a hematological malignancy or autoimmune disease at the time of collection.

[0032] By "individuals suffering from or at risk of suffering from age-related cognitive impairment" is meant individuals who have approximately more than 50% of the life expectancy, e.g., more than 60%, e.g., more than 70%, e.g., more than 75%, 80%, 85%, 90%, 95% or even more than 99% of the life expectancy. The age of the individual depends on the species. Thus, the ratio is based on the expected life expectancy of the species. For example, in humans, such individuals include individuals aged 50 years or older, e.g., 60 years or older, 70 years or older, 80 years or older, 90 years or older, and usually less than 100 years, e.g., 90 years, i.e., between about 50 and 100 years, e.g., 50...55...60...65...70...75...80...85...90...95...100 years or older, any age between 50 and 100 years, who have not yet shown symptoms of an age-related condition, e.g., cognitive impairment, as described further below. and usually less than 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; individuals of any age who suffer from cognitive impairment due to an age-related disease as described further below; and individuals of any age who have been diagnosed with an age-related disease that usually involves cognitive impairment, but in which the individual has not yet begun to show symptoms of cognitive impairment. The corresponding ages for non-human subjects are known and are intended to apply herein.

[0033] As used herein, "treatment" refers to either (i) prevention of a disease or disorder, or (ii) reduction or elimination of symptoms of a disease or disorder. Treatment may be achieved prophylactically (prior to the onset of the disease) or therapeutically (subsequent to the onset of the disease). The effect may be preventative in that the disease or its symptoms are completely or partially prevented, and / or may be therapeutic in that the disease and / or the deleterious effects caused by the disease are partially or completely cured. Thus, the term "treatment" as used herein covers the treatment of age-related diseases or disorders in mammals, including (a) preventing the disease from occurring in a subject who may be susceptible to the disease but has not yet been diagnosed as having it, b) inhibiting the disease, i.e., arresting 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 altering gene expression, rejuvenating tissues or organs, etc. The therapeutic agent 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 alleviates undesirable clinical symptoms in the patient. Such treatment may be performed prior to complete loss of function in the affected tissue. The subject therapies may be administered during, and in some cases after, the symptomatic stage of the disease.

[0034] In some embodiments, the age-related condition to be treated is an age-related impairment of cognitive ability in an individual. By cognitive ability or "cognition" is meant mental functions including attention and concentration, mastering complex tasks and concepts, memory (acquiring, retaining, and retrieving new information in short and / or long terms), information processing (acting on information gathered by the senses), visuospatial functions (visual perception using mental imagery, copy-drawing, constructing objects or shapes, depth perception), producing and understanding language, speech fluency (word finding), problem solving, decision making, and executive functions (planning and prioritizing). By "cognitive decline" is meant the progressive decline in one or more of these abilities, such as the decline in memory, language, thinking, judgment, etc. By "impairment in cognitive ability" and "cognitive impairment" is meant a decline in cognitive ability compared to a healthy individual, e.g., an age-matched healthy individual, or compared to the individual's ability 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. By "age-related cognitive impairment" is meant, for example, cognitive impairment associated with the natural aging process, e.g., impairment of cognitive ability normally associated with aging, including mild cognitive impairment (MCI), and age-related disorders that are disorders that are seen with increasing frequency with increasing aging, e.g., cognitive impairment associated with neurodegenerative conditions, e.g., Alzheimer's disease, Parkinson's disease, frontotemporal dementia, Huntington's disease, amyotrophic lateral sclerosis, multiple sclerosis, glaucoma, myotonic dystrophy, vascular dementia, etc.

[0035] In some embodiments, the age-related condition to be treated is an age-related impairment of motor skills in an individual. By motor skills, we mean the motor process, including the ability to perform complex muscle and nerve activities that produce movements, such as fine motor skills (e.g., writing, tying shoelaces) that produce small and precise movements, and gross motor skills (e.g., walking, running, kicking) for larger movements. By "motor decline" we mean the progressive decline in one or more of these abilities, such as fine or gross motor skills. By "impaired movement" and "motor disorder" we mean the decline in motor abilities / skills compared to healthy individuals, e.g., age-matched healthy individuals, or compared to the abilities of that individual at a previous time point, e.g., 2 weeks, 1 month, 2 months, 3 months, 6 months, 1 year, 2 years, 5 years, or 10 years or more ago. By "age-related movement disorder" is meant, for example, movement disorders or decline associated with the natural aging process, as well as movement disorders or decline associated with aging, i.e. disorders that are seen with increasing frequency with increasing aging, e.g., movement disorders or decline associated with neurodegenerative conditions such as Parkinson's disease, amyotrophic lateral sclerosis, and the like, that are typically associated with aging.

[0036] In some embodiments, the age-related condition to be treated is an age-related increase in neuroinflammation in an individual. By "neuroinflammation" is meant the biochemical and cellular response of the nervous system to injury, infection, or neurodegenerative disease. Such response is directed to reducing triggers by involving the immune system of the central nervous system to defend against potential harm. Neurodegeneration occurs in the central nervous system and is characterized by the loss of neuronal structure and function. Neuroinflammatory diseases or conditions or diseases associated with neuroinflammation include, by way of example and without limitation, neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, multiple sclerosis, and the like.

[0037] Blood products containing plasma components In practicing the subject methods, a blood product comprising plasma is administered to an individual in need thereof, e.g., an individual suffering from or at risk of suffering from cognitive or movement disorders, neuroinflammation, and / or age-related dementia. As such, methods according to embodiments of the present invention include administering a blood product comprising a plasma component derived from an individual ("donor individual" or "donor") to an individual ("recipient individual" or "recipient") at least at risk of suffering from or suffering from cognitive or movement disorders, neuroinflammation, neurodegenerative and / or age-related dementia. By "blood product comprising plasma component" is meant a product derived from blood (e.g., whole blood, blood plasma or a fraction thereof) comprising plasma. The term "plasma" is used in the conventional sense to refer to the straw-colored / light yellow liquid component of blood, which is composed of approximately 92% water, 7% proteins such as, e.g., albumin, gamma globulin, antihemophilic factors and other clotting factors, and 1% mineral salts, sugars, fats, hormones and vitamins. Non-limiting examples of plasma-containing blood products suitable for use in the subject methods include whole blood treated with an anticoagulant (e.g., EDTA, citrate, oxalate, heparin, etc.), blood products made by filtering whole blood to remove leukocytes ("leukocyte depletion"), blood products consisting of plasma derived from plasma exchange or from apheresis, fresh frozen plasma, blood products consisting essentially of purified plasma, and blood products consisting essentially of plasma fractions. In some cases, the plasma product employed is a non-whole blood plasma product, by which is meant that the product is not whole blood, such that it lacks one or more components found in whole blood, such as, for example, 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, where in such examples the cellular components may be 5% by volume, e.g., 1% or less, including 0.5% or less, where in some cases the acellular plasma fraction is a composition completely devoid of cells, i.e., they are cell-free.

[0038] Collection of blood products containing plasma components The embodiments of the methods described herein include administration of blood products containing plasma components that can be derived from donors, including human volunteers. The term "human-derived" can refer to such products. Methods for harvesting blood products, including plasma, from donors are well known in the art (see, for example, AABB TECHNICAL MANUAL, (Mark A. Fung, et al., eds., 18th ed. 2014), which is incorporated herein by reference).

[0039] In one embodiment, the blood donation is obtained by venipuncture. In another embodiment, the venipuncture is a single venipuncture. In another embodiment, volume replacement with saline is not employed. In a preferred embodiment, the process of plasma exchange is used to obtain the blood product comprising plasma. Plasma exchange can include removing a weight-adjusted volume of plasma and returning the cellular components to the donor. In a preferred embodiment, sodium citrate is used during plasma exchange to prevent clotting of cells. The volume of plasma taken from the donor is preferably 690-880 mL after citrate administration, and is preferably adjusted for the weight of the donor.

[0040] 3. Plasma Fraction During World War II, when soldiers lost large amounts of blood, a need arose for a stable plasma expander that could be employed on the battlefield. As a result, methods were developed to prepare freeze-dried plasma. However, the use of freeze-dried plasma was difficult in combat situations because reconstitution required sterile water. As an alternative, Dr. E. J. Cohn proposed that albumin could be used to prepare a ready-to-use stable solution that could be immediately introduced to treat shock (see Johan, Current Approaches to the Preparation of Plasma Fractions in (Biotechnology of Blood) 165 (Jack Goldstein ed., 1st ed. 1991)). Dr. Cohn's procedure for purifying plasma fractions utilized cold ethanol for its denaturing effect, but employed changes in pH and temperature to achieve separation.

[0041] An embodiment of the method described herein includes administering a plasma fraction to a subject. Fractionation is a process in which specific protein subsets are separated from plasma. Fractionation techniques are known in the art and rely on a process developed by Cohn, et al. during 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). Several steps are involved in this process, each step including a specific ethanol concentration and pH, temperature and osmolarity shifts that result in selective protein precipitation. The precipitate is also separated via centrifugation or sedimentation. The original "Cohn fractionation process" involved the separation of proteins via precipitation into five fractions named fraction I, fraction II+III, fraction IV-1, fraction IV-4 and fraction V. Albumin was the originally identified end-point (fraction V) product of this process.According to embodiments of the invention, each fraction (or eluate from a previous 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. Patent Nos. 3,869,431, 5,110,907, 5,219,995, 7,531,513, and 8,772,461, all of which are incorporated herein by reference). Adjustments of the above experimental parameters can be made to obtain specific protein fractions.

[0042] More recently, fractionation has reached further complexity and as such includes additional embodiments of the present invention. This recent increase in complexity has occurred through the introduction of chromatography resulting in the isolation of new proteins from existing fractions such as cryoprecipitate, undercooled plasma, and Cohn fractions, increased IgG recovery by integrating chromatography with ethanol fractionation, and viral reduction / inactivation / removal (Id.). Anion exchange chromatography can be utilized to capture proteins at physiological pH and ionic strength. This preserves the functional activity of the protein and / or protein fraction. Heparin and monoclonal antibodies are also used in affinity chromatography. Those skilled in the art will recognize that the parameters described above may be adjusted to obtain fractions containing the specifically desired plasma protein.

[0043] In an embodiment of the invention, blood plasma is fractionated in an industrial environment. Frozen plasma is thawed at 1°C to 4°C. Continuous refrigerated centrifugation is applied to the thawed plasma and cryoprecipitate is isolated. The collected cryoprecipitate is frozen and stored at -30°C or lower. The cryoprecipitate-poor ("cryopoor") plasma is immediately processed for capture (e.g., via primary chromatography) of protease inhibitors such as antithrombin and C1 esterase inhibitor as well as labile coagulation factors such as factor IX complex and its components. Continuous centrifugation and precipitation isolation can be applied in subsequent steps. 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, 5,288,853, and U.S. Patent Application Nos. 20140343255 and 20150343025, the disclosures of which are incorporated herein by reference in their entirety.

[0044] In an embodiment of the invention, the plasma fraction may include a plasma fraction containing a substantial concentration of albumin. In another embodiment of the invention, the plasma fraction may include a plasma fraction containing a substantial concentration of IgG or intravenous immunoglobulin (IGIV) (e.g., Gamunex-C®). In another embodiment of the invention, the plasma fraction may include an IGIV plasma fraction, such as Gamunex-C®, that has been substantially depleted of immunoglobulin (IgG) by methods 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 an additional embodiment, the blood plasma fraction may have substantially all clotting factors removed to retain the efficacy of the fraction with reduced risk of thrombosis. For example, the plasma fraction may be a plasma fraction as described in U.S. Patent No. 62 / 376,529, filed August 18, 2016, the disclosure of which is incorporated by reference in its entirety.

[0045] 4. Albumin preparations To those skilled in the art, there are two general categories of albumin plasma preparations ("APPs"): plasma protein fractions ("PPFs") and human albumin solutions ("HASs"). PPFs are derived from a process that produces a higher yield than HASs, but have a lower minimum albumin purity than HASs (>83% for PPFs and >95% for HASs). (Production of Human Albumin Solution: a continually developing colloid, P. Matejtschuk, et al., British J. of Anaesthesia, 85(6):887-95, at 888 (2000)). In some cases, PPFs have an albumin purity between 83%-95%, or alternatively between 83%-96%. Albumin purity can be determined by electrophoresis or other quantitative assays, such as, for example, by mass spectrometry. Additionally, some have noted that PPFs have shortcomings due to the presence of protein "contaminants" such as, for example, PKA (Id.). As a result, PPF preparations have lost popularity as albumin plasma preparations and have even been delisted from the pharmacopoeias of certain countries (Id.). Contrary to these concerns, the present invention makes beneficial use of these "contaminants". In addition to alpha, beta and gamma globulins as well as PKA as discussed above, the methods of the present invention utilize additional proteins or other factors within the "contaminants" that promote processes such as, for example, neurogenesis, neuronal survival, improved cognitive or motor function, and reduced neuroinflammation.

[0046] Those of skill in the art will recognize that there are or have been several commercially available sources of PPF (e.g., "commercially available PPF formulations"). These include Plasma-Plex™ PPF (Armour Pharmaceutical Co., Tarrytown, NY), Plasmanate™ PPF (Grifols, Clayton, NC), Plasmatein™ (Alpha Therapeutics, Los Angeles, CA), and Protectate™ PPF (Baxter Labs, Inc. Deerfield, IL).

[0047] Those of skill in the art will also recognize that there are or have been several commercially available sources of HAS ("commercially available HAS formulations"). These include Albuminar™ (CSL Behring), AlbuRx™ (CSL Behring), Albutein™ (Grifols, Clayton, NC), Buminate™ (Baxatla, Inc., Bannockburn, IL), Flexbumin™ (Baxatla, Inc., Bannockburn, IL), and Plasbumin™ (Grifols, Clayton, NC).

[0048] a. Plasma Protein Fraction (Human) (PPF) According to the U.S. Food and Drug Administration ("FDA"), "Plasma Protein Fraction (Human)" or PPF is the proper 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, incorporated herein by reference). The source material for PPF is plasma recovered from whole blood prepared as prescribed in 21 CFR 640.1-640.5, incorporated herein by reference, or source plasma prepared as prescribed in 21 CFR 640.60-640.76, incorporated herein by reference.

[0049] The PPF is reviewed to determine whether it meets the following criteria according to 21 CFR 640.92, which is incorporated herein by reference: (a) the final formulation should be a 5.0±0.30 percent solution of the protein; and (b) The total protein in the final product should consist of at least 83 percent albumin and not more than 17 percent globulins. Not more than 1 percent of the total protein should be gamma globulins. The protein composition will 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.

[0050] As used herein, "plasma protein fraction" or "PPF" refers to a sterile solution of proteins composed of albumin and globulins derived from human plasma, with an albumin content of at least 83%, with no more than 17% globulins (including alpha 1, alpha 2, beta and gamma globulins) and other plasma proteins, and with no more than 1% gamma globulins 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 can also refer to a solid form with 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)).

[0051] B. Albumin (human) (HAS) According to the FDA, "Albumin (Human)" (also referred to herein as "HAS") is the proper name for a drug product defined as a "sterile solution of albumin derived from human plasma" (Code of Federal Regulations 21 CFR 640.80, incorporated herein by reference). The source material for Albumin (Human) is plasma recovered from whole blood prepared as prescribed in 21 CFR 640.1-640.5, incorporated herein by reference, or source plasma prepared as prescribed in 21 CFR 640.60-640.76, incorporated herein by reference. Other requirements for Albumin (Human) are listed in 21 CFR 640.80-640.84, incorporated herein by reference.

[0052] Albumin (Human) is examined to determine that it meets the following criteria per 21 CFR 640.82: (a) Protein Concentration. The final formulation should conform to one of the following concentrations of protein in solution: 4.0±0.25 percent, 5.0±0.30 percent, 20.0±1.2 percent, and 25.0±1.5 percent. (b) Protein Composition.--At least 96 percent of the total protein in the final product should be albumin, as determined by methods approved for each manufacturer by the Director of the Center for Biologics Evaluation and Research of the Food and Drug Administration.

[0053] 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 5% or less globulins (including α1, α2, β and γ globulins) and other plasma proteins. HAS can also refer to a solid form with a similar composition when suspended in a solvent. The total globulin fraction can be determined by subtracting albumin from the total protein.

[0054] As can be appreciated by those skilled in the art, the PPF fraction and the HAS fraction can be lyophilized or in other solid forms. Such preparations can be used with appropriate additives to make, for example, tablets, powders, granules, or capsules. The 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 aliphatic acid glycerides, esters of higher aliphatic acids, or propylene glycol, if desired, with conventional additives such as, for example, solubilizers, isotonicity agents, suspending agents, emulsifiers, stabilizers, and preservatives.

[0055] 5. Depleted coagulation factor fractions Another embodiment of the invention uses blood plasma fractions from which substantially all clotting factors have been removed to retain the efficacy of the fraction with reduced risk of thrombosis. Conveniently, the blood product can be derived from a young donor or a pool of young donors and can be made to lack IgM to provide an ABO-compatible young blood product. Currently, the transfused plasma is matched for ABO blood type, since the presence of naturally occurring antibodies against A and B antigens can result in transfusion reactions. IgM is believed to be involved in transfusion reactions when patients are given ABO-incompatible plasma. Removal of IgM from blood products or blood fractions helps to eliminate transfusion reactions in subjects receiving the blood products or blood plasma fractions of the invention.

[0056] Thus, in one embodiment, the present invention relates to a method of treating or preventing age-related conditions, such as, for example, cognitive or motor disorders, neuroinflammation or neurodegeneration, in a subject. The method comprises administering to the subject a blood product or blood fraction derived from whole blood from an individual or a pool of individuals, wherein the blood product or blood fraction is substantially devoid of (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 is substantially devoid of at least one clotting factor and IgM. In certain embodiments, the blood product is substantially devoid of fibrinogen (Factor I). In additional embodiments, the blood product is substantially devoid of 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, which is incorporated herein by reference in its entirety.

[0057] 6. Processing of protein-enriched plasma protein preparations Additional embodiments of the invention use plasma fractions with reduced albumin concentration compared to PPF, but increased amounts of globulins and other plasma proteins (what some refer to as "contaminants"). The embodiments are effectively devoid of all clotting factors, as are PPF, HAS, Eluent I and Eluent II / III. Such plasma fractions are hereafter referred to as "protein-enriched plasma protein preparations." For example, an embodiment of the invention may use a protein-enriched plasma protein preparation composed of 82% albumin and 18% alpha, beta and gamma globulins and other plasma proteins. Another embodiment of the invention may use a protein-enriched plasma protein preparation composed of 81% albumin and 19% alpha, beta and gamma globulins and / or other plasma proteins. Another embodiment of the invention may use a protein-enriched plasma protein preparation composed of 80% albumin and 20% alpha, beta and gamma globulins and / or other plasma proteins. Additional embodiments of the invention may use a plasma protein preparation enriched with proteins composed of 70-79% albumin and 21-30% alpha, beta and gamma globulins and other plasma proteins. Additional embodiments of the invention may use a plasma protein preparation enriched with proteins composed of 60-69% albumin and 31-40% alpha, beta and gamma globulins and other plasma proteins. Additional embodiments of the invention may use a plasma protein preparation enriched with proteins composed of 50-59% albumin and 41-50% alpha, beta and gamma globulins and other plasma proteins. Additional embodiments of the invention may use a plasma protein preparation enriched with proteins composed of 40-49% albumin and 51-60% alpha, beta and gamma globulins and other plasma proteins. Additional embodiments of the invention may use a plasma protein preparation enriched with proteins composed of 30-39% albumin and 61-70% alpha, beta and gamma globulins and other plasma proteins. Additional embodiments of the invention may use a protein-enriched plasma protein preparation composed of 20-29% albumin and 71-80% alpha, beta and gamma globulins and other plasma proteins.Additional embodiments of the invention may use a protein-enriched plasma protein preparation consisting of 10-19% albumin and 81-90% alpha, beta and gamma globulins and other plasma proteins. Additional embodiments of the invention may use a protein-enriched plasma protein preparation consisting of 1-9% albumin and 91-99% alpha, beta and gamma globulins and other plasma proteins. Further embodiments of the invention may use a protein-enriched plasma protein preparation consisting of 0% albumin and 100% alpha, beta and gamma globulins and other plasma proteins.

[0058] The embodiments of the invention described above may also have a total gamma globulin concentration of 1-5%.

[0059] The specific concentrations of proteins in the plasma fractions may be determined using techniques well known to those of skill in the relevant art, including, by way of example and without limitation, electrophoresis, mass spectrometry, ELISA analysis and Western blot analysis.

[0060] 7. Preparation of Plasma Fractions Methods for preparing PPF and other plasma fractions are well known to those skilled in the art. An embodiment of the present invention allows blood used to prepare human plasma protein fractions to be collected in a flask with citrate or anticoagulant citrate dextrose solution for inhibition of coagulation, and then separated into fractions I, II+III, IV, and PPF as disclosed in 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), which is incorporated herein by reference). According to this method, the mixture can be collected at 2-8°C. The plasma can then be subsequently 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 after removal from storage at -20°C.

[0061] Plasma can be separated from fraction I using 8% ethanol at a protein concentration of 5.1 to 5.6 percent at pH 7.2 and a temperature of -2 to -2.5°C. Cold 53.3 percent ethanol (176 mL / L of plasma) can be added with acetate buffer (200 mL of 4 M sodium acetate, 230 mL of glacial acetic acid quantum salts to 1 L with H2O) using a nozzle at a rate of, for example, 450 mL / min while lowering the plasma temperature to -2°C. Fraction I can be separated and removed from the eluate (eluate I) via centrifugation. Fibrinogen can be obtained from fraction I as per methods well known to those skilled in the art.

[0062] Fraction II+III can be separated from Eluate I via adjusting the eluate to 21 percent ethanol with a protein concentration of 4.3 percent at pH 6.8 and a temperature of -6° C. Cold 95 percent ethanol (176 mL / L of Eluate I) can be added with 10 M acetic acid used for pH adjustment, using a nozzle at a rate of, for example, 500 mL / min while lowering the temperature of Eluate I to -6° C. 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.

[0063] Fraction IV-1 can be separated from eluate II+III ("eluate II / III") via adjusting the eluate to 19 percent ethanol at a protein concentration of 3 percent at pH 5.2 and a temperature of -6°C. H2O and 10M acetic acid used for pH adjustment can be added using a nozzle while maintaining eluate II / III at -6°C for 6 hours. Precipitating fraction IV-1 can be allowed to precipitate at -6°C for 6 hours and then separated from the eluate by centrifugation at the same temperature. Stable plasma protein fractions can be recovered from eluate IV-1 via adjusting the ethanol concentration to 30 percent at a protein concentration of 2.5 percent at pH 4.65 and a temperature of -7°C. This can be achieved by adjusting the pH of eluate IV-1 with cold acid-alcohol (2 parts 2M acetic acid and 1 part 95 percent ethanol). Add 170 mL of cold ethanol (95%) for every liter of adjusted eluate IV-1 while maintaining the temperature at -7°C. Precipitating proteins are allowed to precipitate for 36 hours and then removed by centrifugation at -7°C.

[0064] The recovered proteins (stable plasma protein fraction) can be dried (e.g., by lyophilization) to remove alcohol and H2O. The resulting dry powder can be dissolved in sterile distilled water, for example, using 15 liters of water / kg of powder, and adjusting the solution to pH 7.0 with 1 M NaOH. A final concentration of 5 percent protein can be achieved by adding sterile distilled water containing sodium acetyl tryptophanate, sodium caprylate, and NaCl, adjusting to a final concentration of 0.004 M acetyl tryptophanate, 0.004 M caprylate, and 0.112 M NaCl. Finally, the solution can be filtered at 10°C to obtain a clear solution, and then heat treated at 60°C for at least 10 hours for inactivation of pathogens.

[0065] Those skilled in the art will recognize that each of the various fractions and eluates described above may be used in conjunction with the methods of the present invention to treat diseases. For example, and not by way of limitation, Elution I or Elution II / III may be utilized to treat diseases such as cognitive disorders, movement disorders, and neurodegenerative disorders, which are embodiments of the present invention.

[0066] The preceding methods of preparing plasma and plasma protein fractions (PPF) are merely exemplary and comprise merely embodiments of the present invention. Those skilled in the art will recognize that these methods may vary. For example, pH, temperature, and ethanol concentration, among others, may be adjusted to produce different variations of plasma and plasma protein fractions in various embodiments and methods of the present invention. In another example, additional embodiments of the present invention contemplate the use of nanofiltration to remove / inactivate pathogens from plasma and plasma protein fractions.

[0067] Additional embodiments of the present invention contemplate methods and compositions using and / or including additional plasma fractions. For example, the present invention demonstrates, among other things, that a particular concentration of albumin is not critical for improving cognitive or motor activity. Thus, fractions with reduced albumin concentrations, e.g., those fractions having less than 83% albumin, are contemplated by the present invention.

[0068] 8.Treatment Aspects of the methods of the invention described herein include treating a subject with a blood product comprising plasma, e.g., a blood plasma fraction as described above. An embodiment includes treating a human subject with a blood product comprising plasma. One skilled in the art will recognize that methods of treating a subject with a blood product comprising plasma are recognized in the art. By way of example and not limitation, one embodiment of the methods of the invention described herein comprises administering fresh frozen plasma to a subject for the treatment of cognitive or motor disorders, neuroinflammation, neurodegenerative and / or age-related dementia. In one embodiment, the blood product comprising plasma is administered immediately, e.g., within 12-48 hours, after collection from a donor to an individual suffering from or at risk of cognitive or motor disorders, neuroinflammation, neurodegenerative and / or age-related dementia. In such an example, the formulation may be stored under refrigeration, e.g., at 0-10°C. In another embodiment, the fresh frozen plasma is stored frozen (low temperature) at -18°C or below. Prior to administration, the fresh frozen plasma is thawed and, once thawed, is administered to the subject 60-75 minutes after the thawing process begins. Each subject preferably receives a single unit of fresh frozen plasma (200-250 mL), the fresh frozen plasma being derived from a donor in a pre-determined age range. In one embodiment of the invention, the fresh frozen plasma is provided by (derived from) a younger individual. In another embodiment of the invention, the fresh frozen plasma is provided by (derived from) a donor of the same sex. In another embodiment of the invention, the fresh frozen plasma is provided by (derived from) a donor in the age range between 18-22 years.

[0069] In an embodiment of the invention, blood products including plasma are tested after donation for blood type. In another embodiment of the invention, blood products including plasma are tested for infectious disease pathogens such as, for example, HIV I and II, HBV, HCV, HTLV I and II, anti-HBc as per the requirements and recommendations of 21 CFR 640.33 contained in FDA guidance documents.

[0070] In yet another embodiment of the invention, the subject is treated with a plasma fraction. In an embodiment of the invention, the plasma fraction is PPF or HAS. In a further embodiment of the invention, the plasma fraction is one of the commercially available PPF formulations or commercially available HAS formulations. In another embodiment of the invention, the plasma fraction is PPF or HAS derived from individuals of a certain age range, e.g., a pool of young individuals, or is a modified PPF or HAS fraction that has been subjected to additional fractionation or processing (e.g., PPF or HAS from which one or more specific proteins have been partially or substantially removed). In another embodiment of the invention, the plasma fraction is an IGIV plasma fraction that has been substantially depleted of immunoglobulins (IgG). A blood fraction having a particular protein such as IgG "substantially depleted" or "substantially removed" refers to a blood fraction that contains less than about 50% of the amount present in a reference formulation or whole blood plasma, e.g., less than 45%, 40%, 35%, 30%, 25%, 20%, 15%, 5%, 4%, 3%, 2%, 1%, 0.5%, .25%, .1%, an undetectable level, or any integer value between these values, as measured using standard assays well known in the art.

[0071] 9. Administration Aspects of the methods of the invention described herein include, for example, treatment of a subject with a blood product comprising plasma, e.g., blood plasma or plasma fraction, as described above. Embodiments include treatment of a human subject with a blood product comprising plasma. One skilled in the art will recognize that methods of treatment of a subject with a blood product comprising plasma are recognized in the art. By way of example and not limitation, one embodiment of the methods of the invention described herein comprises administering fresh frozen plasma to a subject for the treatment and / or prevention of cognitive or motor disorders, neuroinflammation, neurodegenerative and / or age-related dementia. In one embodiment, the blood product comprising plasma is administered immediately, e.g., within 12-48 hours, after collection from a donor to an individual suffering from or at risk of cognitive or motor disorders, neuroinflammation, neurodegenerative and / or age-related dementia. In such an example, the formulation may be stored under refrigeration, e.g., at 0-10°C. In another embodiment, the fresh frozen plasma is stored frozen (low temperature) at -18°C or below. Prior to administration, the fresh frozen plasma is thawed and, once thawed, is administered to the subject 60-75 minutes after the thawing process begins. Each subject preferably receives a single unit of fresh frozen plasma (200-250 mL), the fresh frozen plasma being derived from a donor in a pre-determined age range. In one embodiment of the invention, the fresh frozen plasma is provided by (derived from) a younger individual. In another embodiment of the invention, the fresh frozen plasma is provided by (derived from) a donor of the same sex. In another embodiment of the invention, the fresh frozen plasma is provided by (derived from) a donor in the age range between 18-22 years.

[0072] In an embodiment of the invention, blood products including plasma are tested after donation for blood type. In another embodiment of the invention, blood products including plasma are tested for infectious disease pathogens such as, for example, HIV I and II, HBV, HCV, HTLV I and II, anti-HBc as per the requirements and recommendations of 21 CFR 640.33 contained in FDA guidance documents.

[0073] In yet another embodiment of the invention, the subject is treated with a plasma fraction. In an embodiment of the invention, the plasma fraction is PPF or HAS. In a further embodiment of the invention, the plasma fraction is one of the commercially available PPF formulations or commercially available HAS formulations. In another embodiment of the invention, the plasma fraction is PPF or HAS derived from individuals of a certain age range, e.g., a pool of young individuals, or is a modified PPF or HAS fraction that has been subjected to additional fractionation or processing (e.g., PPF or HAS from which one or more specific proteins have been partially or substantially removed). In another embodiment of the invention, the plasma fraction is an IGIV plasma fraction that has been substantially depleted of immunoglobulins (IgG). A blood fraction having a particular protein such as IgG "substantially depleted" or "substantially removed" refers to a blood fraction that contains less than about 50% of the amount present in a reference formulation or whole blood plasma, e.g., less than 45%, 40%, 35%, 30%, 25%, 20%, 15%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.25%, 0.1%, undetectable levels, or any integer value between these values, as measured using standard assays well known in the art.

[0074] An embodiment of the present invention includes treating a subject diagnosed with cognitive or motor disorders, neurodegeneration or neuroinflammation by administering an effective amount of blood plasma or plasma fraction to the subject. Another embodiment of the present invention includes administering an effective amount of blood plasma or plasma fraction and then monitoring the subject for improvement of cognitive or motor disorders, or reduction of neuroinflammation, or increase in neurogenesis. Another embodiment of the present invention includes treating a subject diagnosed with cognitive or motor disorders, neurodegeneration or neuroinflammation by administering an effective amount of blood plasma or plasma fraction to the subject, where the blood plasma or plasma fraction is administered in a manner that results in improvement of cognitive or motor disorders, reduction of neuroinflammation, or improvement of neurogenesis after reaching the mean or median half-life of the blood plasma protein or plasma fraction protein compared to the most recently administered dose (referred to herein as "pulsed dosing" or "pulsed"). Another embodiment of the invention includes administering blood plasma or plasma fractions via a dosing regimen of at least two consecutive days and monitoring the subject for improved cognitive or motor function, reduced neuroinflammation, or improved neurogenesis at least three days after the date of the last administration. Further embodiments of the invention include administering blood plasma or plasma fractions via a dosing regimen of at least three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, or fourteen consecutive days and monitoring the subject for improved cognitive or motor function, reduced neuroinflammation, or improved neurogenesis at least three days after the date of the last administration. Yet another embodiment of the invention includes administering blood plasma or plasma fractions via a dosing regimen of at least two consecutive days and after the date of the last administration and monitoring for improved cognitive or motor function, reduced neuroinflammation, or improved neurogenesis past the time the average half-life of the protein in the blood plasma or plasma fraction is reached. Another embodiment of the invention involves administering blood plasma or plasma fractions via a dosing regimen of 2 to 14 non-consecutive days, each interval between doses being optionally 0 to 3 days.

[0075] In some cases, pulsatile dosing according to the invention includes administration of a first set of doses, e.g., as described above, followed by a dosing-free period, e.g., a "drug-free period", followed by administration of another dose or set of doses. The duration of this "drug-free period" may vary, but in some embodiments is 7 days or more, e.g., 10 days or more, including 14 days or more, and in some cases the dosing-free period ranges from 15 to 365 days, e.g., 30-90 days and 30-60 days. As such, method embodiments include non-chronic (i.e., non-continuous) dosing, e.g., non-chronic administration of blood plasma products. In some embodiments, the pattern of pulsatile dosing followed by a drug-free period is repeated as many times as desired, and in some cases, the pattern is continued for one year or more, e.g., two years or more, up to and including the life of the subject. Another embodiment of the invention includes administering blood plasma or plasma fractions via a continuous five-day dosing regimen with a 2-3 day drug-free period followed by 2-14 consecutive days of dosing.

[0076] Biochemically, by "effective amount" or "effective dose" of an active agent is meant an amount of active agent that inhibits, antagonizes, reduces, mitigates, or arrests the progression of cognitive or motor impairment, neuroinflammation, neurodegeneration, or age-related dementia by about 20% or more, e.g., 30% or more, 40% or more, or 50% or more, in some cases, 60% or more, 70% or more, 80% or more, or 90% or more, in some cases, about 100%, i.e., to a negligible amount, and in some cases, reverses it.

[0077] 10. Plasma Protein Fractions In carrying out the method of the present invention, a plasma fraction is administered to a subject.In an embodiment, the plasma fraction is a plasma protein fraction (PPF).In a further embodiment, the PPF is selected from commercially available PPF preparations.

[0078] In another embodiment, PPF is composed of 88% normal human albumin, 12% alpha and beta globulins, and 1% or less gamma globulins as measured by electrophoresis. Further embodiments of this embodiment used to practice the methods of the invention include, for example, an embodiment as a 5% solution of PPF buffered with sodium carbonate and stabilized with 0.004M sodium caprylate and 0.004M acetyl tryptophan. Additional formulations may be utilized in practicing the methods of the invention, including those that vary the percentage of PPF in solution (e.g., about 1% to about 10%, about 10% to about 20%, about 20% to 25%, about 25% to 30%) as well as the concentrations of solvents and stabilizers.

[0079] 11. Plasma fractions of specific donor ages Additional embodiments of the present invention include administering plasma protein fractions derived from the plasma of individuals of a particular age range.Embodiments include 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 of a particular age range.In yet another embodiment, the average age of the donor is less than that of the subject or is less than the average age of the subject to be treated.

[0080] A particular embodiment of the invention involves pooling blood or blood plasma from individuals of a particular age range and fractionating the blood plasma as described above to obtain a plasma protein fraction preparation such as PPF or HAS. In an alternative embodiment of the invention, the plasma protein fraction or a particular plasma protein fraction is obtained from a particular individual that fits into a particular age range.

[0081] 12. Indications The subject methods and blood products and blood fractions, including plasma, are used to treat, including preventing, age-related conditions, such as impairments in an individual's cognitive or motor abilities, including, but not limited to, cognitive impairments, including, but not limited to, age-related dementia, immune conditions, cancer, and physical or functional decline, and motor disorders, such as, but not limited to, Parkinson's disease. For example, individuals suffering from or at risk of developing age-related cognitive or motor disorders, neuroinflammation and / or neurodegeneration that would benefit from treatment with a subject blood product, including plasma, 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 of age, e.g., 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or about 100 years of age, and who are of normal age and have a normal aging. These individuals include individuals suffering from cognitive or motor impairments, neuroinflammation and / or neurodegeneration associated with the natural aging process, e.g., mild cognitive impairment (MCI), and individuals who are 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 usually under 100 years of age, i.e., between about 50 and 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 or motor impairments, neuroinflammation and / or neurodegeneration. Examples of cognitive or motor, neuroinflammation and / or neurodegeneration disorders that are due to natural aging include:

[0082] (a) Mild cognitive impairment (MCI) Mild cognitive impairment is a mild disruption of cognition that manifests as problems with memory or other mental functions, such as planning, following instructions, or making decisions, that deteriorate over time, while overall mental function and daily activities are intact.Thus, although significant neuronal death does not usually occur, neurons in the aging brain are susceptible to sublethal age-related changes in synaptic structure, synaptic integrity, and synaptic molecular processing, all of which impair cognitive function.

[0083] For example, subjects suffering from or at risk of developing age-related cognitive impairment who would benefit from treatment with a subject plasma-containing blood product or blood fraction according to the methods disclosed herein include subjects of any age who suffer from cognitive impairment due to an age-related disorder, and also subjects of any age who have been diagnosed with an age-related disorder that is usually accompanied by cognitive impairment, where the individual has not yet begun to show symptoms of the cognitive impairment.

[0084] (b) Alzheimer's disease Alzheimer's disease is a progressive and unstoppable loss of cognitive function associated with an excessive number of senile plaques in the cerebral cortex and subcortical gray matter that contain beta-amyloid and neurofibrillary tangles composed of tau protein. The common form affects people over 60 years of age, and its incidence increases with advancing age. It accounts for more than 65% of dementia in the elderly.

[0085] The cause of Alzheimer's disease is unknown. The disease runs in families in approximately 15-20% of cases. The remaining so-called sporadic cases have some genetic determinant. The disease has an autosomal dominant inheritance pattern in most early-onset cases and some late-onset cases, but with variable later-life penetrance. Environmental factors are the focus of active investigation.

[0086] During the course of the disease, synapses and ultimately neurons are lost within 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 several regions of the brain (parietal and temporal lobes in early stage disease, prefrontal cortex in later stage disease). Senile plaques or senile plaques (composed of neurites, astrocytes, and glial cells around an amyloid core) and neurofibrillary tangles (composed of paired helical filaments) play a role in the pathogenesis of Alzheimer's disease. Senile plaques and neurofibrillary tangles occur during normal aging but are much more frequent in people with Alzheimer's disease.

[0087] (c) Parkinson's disease Parkinson's disease (PD) is an idiopathic, slowly progressive degenerative CNS disorder characterized by slow and reduced movement (bradykinesia), muscle stiffness, resting tremor (dystonia), muscle paralysis, and postural instability. Originally considered primarily a movement disorder, PD is now recognized to also cause depression and emotional changes. PD can 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. PD is characterized by symptoms related to reduced motor function that usually precede those associated with cognitive impairment, which aid in the diagnosis of the disease.

[0088] In early Parkinson's disease, pigmented neurons of the substantia nigra, locus coeruleus, and other brainstem dopaminergic cell groups degenerate. The cause is unknown. Loss of substantia nigra neurons projecting to the caudate and nucleus medialis results in depletion of the neurotransmitter dopamine in these areas. Onset is generally after age 40, with an increased incidence in older age groups.

[0089] Parkinson's disease is newly diagnosed in approximately 60,000 Americans each year and currently affects approximately 1 million Americans. Although PD itself is not fatal, its complications are the 14th leading cause of death in the U.S. Currently, PD cannot be cured, and treatments are generally prescribed to manage symptoms and, in late-stage, severe cases, surgery.

[0090] Treatment options for PD include the administration of medications to help manage movement disorders. These options increase or replace dopamine, a neurotransmitter that is low in the brain in PD patients. Such drugs include carbidopa / levodopa (to make more dopamine in the brain), apomorphine, pramipexole, ropinirole, and rotigotine (dopamine agonists), selegiline and rasagiline (MAO-B inhibitors that prevent the breakdown of dopamine), entacapone and tolcapone (catechol-O-methyltransferase [COMT] inhibitors that make more levodopa available in the brain), benztropine and trihexyphenidyl (anticholinergics), and amantadine (to manage tremors and stiffness). Exercise stress / physical therapy is also commonly prescribed to help maintain physical and mental function.

[0091] However, current treatment options treat the symptoms of PD but are not curative and do not prevent disease progression. Furthermore, current drugs tend to lose efficacy in late-stage PD. Levodopa, the most commonly prescribed medication, generally produces adverse effects within 5-10 years of starting the drug. These adverse effects can be severe, resulting in motor fluctuations and unpredictable swings in motor control between doses, as well as spasms / twitching (dyskinesias) that are difficult to manage and even disabling, similar to the symptoms of PD itself. Thus, there remains a need for new therapies with new mechanisms of action that can be administered with or in combination with current PD drugs.

[0092] (d) Parkinsonism Secondary parkinsonism (also called atypical parkinsonism or parkinson plus) 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 antipsychotics or reserpine, which produce parkinsonism by blocking dopamine receptors. Less common causes include carbon monoxide or manganese poisoning, hydrocephalus, structural lesions (tumors, infarcts affecting the midbrain or basal ganglia), subdural hematomas, and degenerative diseases including nigrostriatal degeneration. Certain diseases such as progressive supranuclear palsy (PSP), multiple system atrophy (MSA), corticobasal degeneration (CBD) and dementia with Lewy bodies (DLB) may be classified as "parkinsonism" because they may present with parkinsonism symptoms before the presenting symptoms required for a specific diagnosis can be constituted.

[0093] (e) 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 dementias. Symptoms are classified into three groups based on the function of the frontal and temporal lobes affected.

[0094] Symptom-specific behavioral variants of FTD (bvFTD) include fatigue and loss of spontaneity in one hand and disinhibition in the other, disruption of speech fluency due to difficulties in articulation, progressive non-verbal aphasia (PNFA) in which phonological and / or syntactic errors are observed but word comprehension is preserved, and semantic dementia (SD) in which patients remain fluent with normal phonology and syntax but have increasing difficulties with naming and word comprehension. Other cognitive symptoms common to all FTD patients include impairments in executive function and concentration. Other cognitive abilities, including perception, spatial abilities, memory, and application, remain intact. FTD can be diagnosed by structural MRI scans showing atrophy of the frontal and / or anterior temporal lobes.

[0095] There are many forms of FTD, any of which may be treated or prevented using the subject 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 non-verbal areas of ability. SD patients often present with word-finding difficulties. Clinical signs include fluent aphasia, amnesic aphasia, impaired comprehension of word meanings, and associated visual agnosia (inability to match semantically related pictures or objects). As the disease progresses, cases are described as "pure" semantic dementia with fewer late-onset behavioral symptoms, but often with behavioral and personality changes similar to those seen in frontotemporal dementia. Structural MRI imaging analysis shows a characteristic pattern of atrophy in the temporal lobes (left predominant), with greater involvement of the inferior than the superior, and greater atrophy of the anterior than the posterior temporal lobe.

[0096] As another example, another form of frontotemporal dementia is Pick's disease (PiD, also PcD). The defining feature of the disease is the accumulation of tau protein in neurons, depositing silver-staining globular aggregates known as "Pick bodies". Symptoms include loss of speech (aphasia) and dementia. Patients with orbitofrontal insufficiency can become violent and socially inept. They may also exhibit furtive movements and compulsive or repetitive stereotyped behaviors. Patients with dorsomedial or dorsolateral frontal lobe insufficiency may also exhibit lack of interest, anhedonia, or reduced spontaneous actions. Patients show lack of self-monitoring, abnormal self-awareness, and an inability to fully comprehend meaning. Patients with loss of grey matter in the bilateral posterior orbitofrontal cortex and right anterior insula may also exhibit altered eating behavior, e.g., a pathological sweet tooth. Patients with more localized grey matter loss in the anterior lateral orbitofrontal cortex may develop binge eating. While some symptoms can be alleviated initially, the disease progresses and patients often die within two to ten years.

[0097] (f) Huntington's disease Huntington's disease (HD) is an inherited, progressive neurodegenerative disorder characterized by the occurrence of emotional, behavioral, and psychiatric abnormalities, loss of intellectual or cognitive function, and behavioral abnormalities (movement disorders). Classical signs of HD include chorea - the occurrence of involuntary, rapid, irregular, jerky-like movements that may affect the face, arms, legs, or trunk - as well as cognitive decline, including slow loss of thought processing and acquired intellectual abilities. There may also be impairment of memory, abstract thinking, and judgment, improper perception of time, place, and identity (disorientation), increased agitation, and personality changes (personality disruption). Symptoms usually become evident during the fourth to fifth decades of life, but the age of onset is variable, ranging from early childhood to late adulthood (seventh or eighth decade).

[0098] HD is transmitted in families as an autosomal dominant trait. The disease occurs as a result of an abnormally long sequence or "repeat" of coded instructions within a gene on chromosome 4 (4p16.3). The progressive loss of nervous system function associated with HD results from the loss of neurons in specific regions of the brain, including the basal ganglia and cerebral cortex.

[0099] (g) Amyotrophic lateral sclerosis Amyotrophic lateral sclerosis (ALS) is a rapidly progressive, invariably fatal neurological disease that attacks motor neurons. Signs of muscle weakness and atrophy and anterior horn cell dysfunction are first noticed, most often in the hands and less often in the legs. Site of onset is random and progression is asymmetric. Muscle spasms are common and may precede weakness. Rarely, patients survive for 30 years, but 50% die within 3 years of onset, 20% survive for 5 years, and 10% survive for 10 years.

[0100] Diagnostic features include onset in middle age or old age and progressive, generalized motor involvement in the absence of sensory abnormalities. Nerve conduction velocities are normal until late in the disease. Recent studies have revealed presentation of cognitive impairment as well, particularly declines in immediate verbal memory, visual memory, language, and executive function.

[0101] Reductions in cell body area, number of synapses, and total synaptic length have been reported even in normal-appearing neurons of ALS patients. It has been suggested that continued loss of synapses may lead to functional damage once active zone plasticity reaches its limit. Promoting the formation of new synapses or preventing synapse loss may preserve neuronal function in these patients.

[0102] (h) Multiple sclerosis Multiple sclerosis (MS) is characterized by a variety of symptoms and signs of CNS dysfunction, with remissions and recurrent exacerbations. The most common symptomatic symptoms are sensory disturbances in one or more limbs, one side of the trunk or face, weakness or clumsiness of the legs or hands, or visual disturbances, such as partial blindness and pain in one eye (retrobulbar optic neuritis), dimness or scotoma in the field of vision. Common cognitive disorders include impairments in memory (acquiring, retaining and retrieving new information), attention and concentration (particularly divided attention), information processing, executive function, visuospatial function and verbal fluency. Common early symptoms are ophthalmoplegia resulting in double vision (diplopia), temporary weakness in one or more limbs, mild stiffness or abnormal fatigability of the limbs, mild difficulty walking, difficulty with bladder control, dizziness, and mild emotional lability, all of which indicate diffuse CNS involvement and often occur months or years before the disease is recognized. Extremely high temperatures may accentuate symptoms and signs.

[0103] The course is highly variable and unpredictable, and in most patients it is remittent. Initially, months or years of remission may separate episodes of symptoms, especially when the disease begins with retrobulbar optic neuritis. However, some patients have frequent attacks that can be rapidly incapacitating, and the course can progress rapidly over a short period of time.

[0104] (i) Glaucoma Glaucoma is a common neurodegenerative disease that affects retinal ganglion cells (RGCs). Evidence supports the existence of a compartmentalized degenerative program in synapses and dendrites that contain RGCs. Recent evidence also shows a correlation between cognitive impairment and glaucoma in the elderly (Yochim BP, et al. Prevalence of cognitive impairment, depression, and anxiety symptoms among older adults with glaucoma. J.Glaucoma. 2012; 21(4): 250-254).

[0105] (j) Myotonic dystrophy Myotonic dystrophy (DM) is an autosomal dominant, multisystem disease characterized by dystrophic muscle weakness and myotonia. The molecular defect is an expanded trinucleotide (CTG) repeat in the 3' untranslated region of the myotonin protein kinase gene on chromosome 19q. Symptoms can occur at any age and range in clinical severity. Myotonia is prominent in the hand muscles, and ptosis is common even in mild cases. In severe cases, marked peripheral muscle weakness occurs and is often accompanied by cataracts, premature hair loss, axe-shaped facies, arrhythmias, testicular atrophy, and endocrine abnormalities (e.g., diabetes). Mental retardation is common in the severe congenital form, while age-related decline in frontal and temporal lobe cognitive functions, especially language and executive functions, is observed in the mild adult form of the disease. Severely affected individuals die by their early 50s.

[0106] (k)Dementia Dementia describes a class of diseases with symptoms that affect thinking and social skills significantly enough to interfere with daily functioning. In addition to the dementia seen in the later stages of age-related diseases discussed above, other examples of dementia include vascular dementia and Lewy body dementia, which are described below.

[0107] In vascular dementia or "multi-infarct dementia", cognitive impairment is caused by problems with the blood supply to the brain, usually a series of small strokes, or sometimes one large stroke preceding or followed by other small strokes. Vascular lesions can be the result of diffuse cerebrovascular disease, such as small vessel disease or focal lesions or both. Patients suffering from vascular dementia present with cognitive impairment acutely or subacutely after an acute cerebrovascular event, after which progressive cognitive decline is observed. The associated changes in the brain are not due to AD pathology, but rather to chronic reduction in blood flow in the brain that ultimately results in dementia, but the cognitive impairment, including impairment in language, memory, complex visual processing or executive functions, is similar to that seen in Alzheimer's disease. Neuroimaging by single photon emission computed tomography (SPECT) and positron emission tomography (PET) may be used to confirm the diagnosis of multi-infarct dementia, in conjunction with an evaluation that includes a mental status test.

[0108] Dementia with Lewy bodies (also known by various other names including DLB, dementia with Lewy bodies, diffuse Lewy body disease, cortical Lewy body disease, and Lewy-type age-related dementia) is a form of dementia characterized anatomically by the presence of Lewy bodies (clusters of alpha-synuclein and ubiquitin proteins) in neurons that can be detected by postmortem brain histology. Its main feature is cognitive decline, especially of executive functions. Attention and short-term memory may wax and wane.

[0109] Persistent or recurrent visual hallucinations with vivid, detailed pictures are often the first diagnostic symptom. DLB is often confused with Alzheimer's disease and / or vascular dementia in its early stages, although DLB often has a rapid, acute onset, whereas Alzheimer's disease usually begins quite slowly. DLB symptoms also include motor symptoms similar to those of Parkinson's disease. DLB is differentiated from the dementia that sometimes occurs in Parkinson's disease by the time frame in which the dementia symptoms appear in relation to the Parkinson's symptoms. Parkinson's disease with dementia (POD) is the diagnosis when the onset of dementia occurs more than one year after the onset of Parkinson's disease. DLB is diagnosed when cognitive symptoms begin at the same time as or within one year of Parkinson's symptoms.

[0110] (l) Progressive supranuclear palsy Progressive supranuclear palsy (PSP) is a brain disorder that causes severe and progressive problems with controlling walking and balance, along with complex eye movements and thinking problems. One of the traditional signs of the disease is the inability to look properly, which occurs due to lesions in the area of ​​the brain that coordinates eye movements. Some people describe this effect as blurred vision. Affected people often exhibit mood and behavior changes, including depression and lethargy, as well as progressive mild dementia. The disease's long name indicates that the disease begins slowly, continues to worsen (progressive), and causes weakness (paralysis) by damaging specific parts of the brain above (supranuclear) pea-sized structures called nuclei that control eye movements. PSP was first described as a distinct disease in 1964, when three scientists published a paper that distinguished the condition from Parkinson's disease. It is sometimes called Steele-Richardson-Olszewski syndrome, reflecting the combined name of the scientists who defined the disease. Although PSP progressively worsens, no one has died from PSP itself.

[0111] (m)Ataxia People with ataxia have problems with coordination because the parts of the nervous system that control movement and balance are affected. Ataxia may affect the fingers, hands, arms, legs, body, speech, and eye movements. The word ataxia is often used to describe symptoms of non-coordination that may be related to infection, trauma, other diseases, or degenerative changes in the central nervous system. Ataxia is also used to refer to a group of specific degenerative disorders of the nervous system called hereditary and sporadic ataxias, which are the focus of the American Ataxia Foundation.

[0112] (n) Multiple system atrophy Multiple system atrophy (MSA) is a degenerative neurological disorder. MSA is associated with the degeneration of nerve cells in certain 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.

[0113] 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 presents with some of the same symptoms as Parkinson's disease; however, patients with MSA have a minimal, if any, response to dopamine medications commonly used for Parkinson's disease.

[0114] (o) Flail Frailty syndrome ("frailty") is a geriatric syndrome characterized by functional and physical decline, including reduced mobility, muscle weakness, slow movement, poor endurance, low physical activity, undernutrition, and involuntary weight loss. Such decline is often associated with disease consequences, such as cognitive dysfunction and cancer. However, frailty can also occur in the absence of disease. Individuals with frailty have a high risk of negative outcomes in terms of fractures, accidental falls, disability, comorbidities, and premature mortality (C. Buigues, et al. Effect of a Prebiotic Formulation on Frailty Syndrome: A Randomized, Double-Blind Clinical Trial, Int. J. Mol. Sci. 2016, 17, 932). Furthermore, individuals with frailty have a high incidence of high health care costs. (Ibid.).

[0115] Common symptoms of frailty can be determined by certain types of tests. For example, unintentional weight loss includes a loss of at least 10 lb or more than 5% of body weight in the previous year, muscle wasting can be determined by a decrease in grip strength of at least 20% from baseline (adjusted for gender and BMI), bradykinesia can be based on the time it takes to walk a distance of 15 feet, poor endurance can be determined by a person's own report of feeling exhausted, 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)).

[0116] In some embodiments, the subject methods and compositions are used to slow down the progression of age-related cognitive, motor, neuroinflammation, or other age-related disorders or conditions. In other words, cognitive, motor, neuroinflammation, or other abilities or conditions in an individual decline more slowly following treatment with the disclosed methods than before treatment with the disclosed methods or in the absence of treatment. In some such examples, the subject methods of treatment include measuring the progression of decline in cognitive, motor, neuroinflammation, or other age-related abilities or symptoms after treatment and determining that the progression of decline is reduced. In some such examples, the determination is made by comparing the rate of decline in the individual prior to treatment to a reference, e.g., as determined by previously measuring cognitive, motor, neuroinflammation, or other age-related abilities or conditions at two or more time points prior to administration of the subject blood product.

[0117] The subject methods and compositions are also used in stabilizing the cognitive, motor, neuroinflammatory or other abilities or conditions of an individual, for example, an individual suffering from or at risk of suffering from age-related cognitive decline. For example, the individual may exhibit some age-related cognitive impairment, and the progression of the cognitive impairment observed prior to treatment with the disclosed methods is halted following treatment with the disclosed methods. As another example, the individual may be at risk of developing age-related cognitive decline (e.g., the individual may be over 50 years old or may have been diagnosed with an age-related disorder), and the cognitive performance of the individual is substantially unchanged following treatment with the disclosed methods compared to before treatment with the disclosed methods, i.e., no cognitive decline is detectable.

[0118] The subject methods and compositions can also be used to alleviate cognitive, motor, neuroinflammatory or other age-related disorders in individuals suffering from age-related disorders. In other words, the affected ability is improved in the individual following treatment with the subject methods. For example, the cognitive or motor ability in the individual is increased by 2-fold or more, 5-fold or more, 10-fold or more, 15-fold or more, 20-fold or more, 30-fold or more, or 40-fold or more, including 50-fold or more, 60-fold or more, 70-fold or more, 80-fold or more, 90-fold or more, or 100-fold or more, following treatment with the subject methods, compared to the cognitive or motor ability observed in the individual prior to treatment with the subject methods.

[0119] In some cases, treatment with the subject methods and compositions restores cognitive, motor or other abilities in an individual suffering from age-related cognitive or motor decline, e.g., to the level the individual had when he or she was about 40 years of age or younger. In other words, cognitive or motor impairment is abrogated.

[0120] 13. Methods of diagnosing and monitoring disorders In some cases, among various methods of diagnosing and monitoring disease progression and improvement in cognitive, movement, neurodegenerative and / or neuroinflammatory diseases, the following types of assessments are used alone or in combination with subjects suffering from neurodegenerative diseases as desired. The following types of methods are presented as examples and are not limited to the cited methods. Any convenient method of monitoring disease may be used in practicing the present invention as desired. These methods are also considered by the methods of the present invention.

[0121] (a) General awareness The method embodiment of the present invention further includes a method of monitoring the effect of a drug or treatment on a subject to treat cognitive impairment and / or age-related dementia, the method includes comparing cognitive function before and after treatment. Those skilled in the art recognize that there are well-known methods of evaluating cognitive function. For example, and not by way of limitation, the method may include evaluation of cognitive function based on medical history, family history, physical and neurological examination, clinical examination, and neuropsychological evaluation by a clinician who specializes in dementia and cognitive function. Additional embodiments contemplated by the present invention include, for example, evaluation of consciousness using the Glasgow Coma Scale (EMV), mental status examination including Abbreviated Mental Status Score (AMTS) or Mini Mental Status Examination (MMSE) (Folstein, et al., J. Psychiatr. Res, 1975, 12:1289-198), global evaluation of higher function, for example, estimation of intracranial pressure by fundus examination. In one embodiment, monitoring the effect on cognitive impairment and / or age-related dementia includes an improvement of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 points using the Alzheimer's Disease Assessment Scale-Cognitive Subscale (ADAS-COG).

[0122] In one embodiment, cognitive function may be assessed using tests of the peripheral nervous system including any one of smell, visual field and vision, eye movements and pupils (sympathetic and parasympathetic), facial sensory function, strength of the facial and scapular girdle muscles, hearing, taste, pharyngeal movements and gag reflex, tongue movement (e.g., visual acuity can be assessed by Snellen charts, and a reflex hammer used to assess reflexes including masseter, biceps and triceps tendons, hamstring tendons, ankle reflexes and plantar (i.e., Babinski sign)), muscle strength, muscle tone and signs of stiffness, often on an MRC scale of 1 to 5.

[0123] (b) Parkinson's disease The method embodiments of the present invention further include a method of monitoring the effect of a drug or treatment on a subject to treat a movement disorder, the method comprising comparing motor function before and after treatment. Those skilled in the art will recognize that there are well-known methods of assessing motor function. For example, and not by way of limitation, the method may include assessment of motor function based on medical history, family history, physical and neurological examination, clinical testing and neurodegenerative evaluation by a clinician who specializes in neurodegenerative and movement disorders. Additional embodiments contemplated by the present invention include the adoption of a rating scale as discussed below.

[0124] Several rating scales are available to assess the progression of PD. The most widely used scales include the Unified Parkinson's Disease Rating Scale (UPDRS, introduced in 1987) (J. Rehabil Res. Dev., 2012, 49(8):1269-76), and the Hoehn and Yahr scale (Neruology, 1967, 17(5):427-42). Additional scales include the Schwab and England Activities of Daily Living (ADL) scale as well as the Movement Disorder Society (MDS) updated UPDRS scale (MDS-UPDRS).

[0125] The UPDRS scale assesses 31 items that contribute to three subscales: (1) intellectual activity, behavior, and mood, (2) activities of daily living, and (3) motor examination. The Hoehn and Yahr scale categorizes PD into five stages with careful substages: 0-no signs of disease, 1-symptoms on only one side, 1.5-symptoms on one side but involving the neck and spine, 2-symptoms on both sides and no balance problems, 2.5-mild symptoms on both sides with recovery on "pull test", 3-mild to moderate disease with balance problems, 4-severe disability but with ability to walk or stand independently, and 5-requires wheelchair or is bedridden without support. The Schwab and England scale categorizes PD into several proportions (from 100%-completely independent to 10%-completely dependent).

[0126] General motor function can be assessed using widely used scales including the General Motor Function Scale (GMF), which examines three components: dependency, pain and instability (Aberg, AC, et al. (2003), Disabil. Rehabil. 2003, May, 6; 25(9): 462-72.). Motor function can also be assessed using home monitoring or wearable sensors. For example, gait (speed of movement, variability, leg stiffness) can be sensed by accelerometer, posture (trunk tilt) by gyroscope, leg movements by accelerometer, hand movements by accelerometer and gyroscope, tremor (magnitude, frequency, duration, asymmetry) by accelerometer, falls by accelerometer, freezing of gait by accelerometer, dyskinesia by accelerometer, gyroscope and inertial sensors, bradykinesia (duration and frequency) by accelerometer and gyroscope, and aphasia (pitch) by microphone. (Pastorino, M, et al., Journal of Physics: Conference Series, 450(2013), 012055).

[0127] (c) Multiple sclerosis In addition to improved monitoring of cognition-related symptoms, the progression or improvement of neurodegeneration associated with multiple sclerosis (MS) can be monitored using techniques well known to those skilled in the art. By way of example and not limitation, monitoring can be performed via techniques such as cerebrospinal fluid (CSF) monitoring, magnetic resonance imaging (MRI) to detect the development of lesions and demyelinating plaques, evoked potential testing, and ambulatory monitoring.

[0128] Analysis of the CSF may be performed, for example, via lumbar puncture, to obtain pressure, appearance, and CSF content. Normal values ​​are usually in the following ranges: pressure (70-180 mmH20), appearance clear and colorless, total protein (15-60 mg / 100 mL), IgG 3-12% of total protein, glucose 50-80 mg / 100 mL, cell count 0-5 white blood cells and no red blood cells, and chloride (110-125 mEq / L). Abnormal results may indicate the presence or progression of MS.

[0129] MRI is another technique that may be performed to monitor disease progression and improvement.Typical criteria for monitoring MS with MRI include the appearance of patchy areas of abnormal white matter in the cerebral hemispheres and periventricular areas, lesions in the cervical and thoracic regions of the spinal cord as well as the cerebellum and / or brain stem.

[0130] Evoked potentials may be used to monitor the progression and improvement of MS in a subject. Evoked potentials measure the slowing of electrical stimuli, for example, in visual evoked responses (VER), brainstem auditory evoked responses (BAER), and somatosensory evoked responses (SSER). Abnormal responses serve to indicate that there is a slowing in the velocity of conduction in the central sensory pathways.

[0131] Gait monitoring can also be used to monitor disease progression and improvement in MS subjects. MS is often accompanied by impaired mobility and abnormal gait, some of which is due to fatigue. Monitoring may be performed, for example, with the use of ambulatory monitoring devices worn by the subject. (Moon, Y., et al., Monitoring gait in multiple sclerosis with novel wearable motion sensors, PLOS One, 12(2): e0171346 (2017)).

[0132] (d) Huntington's disease In addition to monitoring improvements in cognition-related symptoms, the progression or improvement of neurodegeneration associated with Huntington's disease (HD) can be monitored using techniques well known to those of skill in the art. By way of example and not limitation, monitoring can be performed via techniques such as motor function, behavior, functional assessment, and imaging analysis.

[0133] Examples of motor functions that may be monitored as indicators of disease progression or improvement include chorea and dystonia, rigidity, bradykinesia, oculomotor dysfunction, and changes in gait / balance. Techniques for performing monitoring of these metrics are well known to those skilled in the art. (See Tang, C, et al., Monitoring Huntington's disease progression through preclinical and early stages, Neurodegener Dis Manag, 2(4):421-35(2012)).

[0134] The psychiatric effects of HD present an opportunity to monitor disease progression and improvement. For example, psychiatric evaluations may be performed to determine whether a subject suffers from depression, irritability, agitation, anxiety, anesthesia, and paranoid psychosis. (Id.)

[0135] Functional assessment may also be employed to monitor disease progression or improvement. Global functional scores have been reported (ibid.), which often result in a loss of 1 point per year in some HD patients.

[0136] MRI or PET may also be employed to monitor disease progression or improvement. For example, in HD there is a loss of striatal-projecting neurons, and changes in the number of these neurons may be monitored in subjects. Techniques for measuring neuronal changes in HD subjects include imaging dopamine D2 receptor binding. (Id.)

[0137] (e) ALS In addition to monitoring improvements in cognition-related symptoms, the progression or improvement of neurodegeneration associated with amyotrophic lateral sclerosis (ALS) can be monitored using techniques well known to those skilled in the art. By way of example and not limitation, monitoring can be performed via techniques such as functional assessment, assessing muscle strength, measuring respiratory function, measuring lower motor neuron (LMN) loss, and measuring upper motor neuron (UMN) dysfunction.

[0138] Functional assessment can be performed using functional measures known to those skilled in the art, such as the ALS Functional Rating Scale (ALSFRS-R), which evaluates symptoms related to bulbar, limb and respiratory function. The rate of change is useful in predicting disease progression or improvement as well as survival. Another assessment measure includes the Combined Assessment of Function and Survival (CAFS), which ranks the subject's clinical outcome by combining survival time with the change in ALSFRS-R. (Simon, NG, et al., Quantifying Disease Progression in Amyotrophic Lateral Sclerosis, Ann. Neurol. 76:643-57 (2014)).

[0139] Muscle strength may be assessed or quantified through the use of composite manual muscle testing (MMT) scores, which involve averaging measurements obtained from several muscle groups using the Medical Research Council (MRC) muscle strength scale. (Id.) Handheld dynamometry (HHD), among other techniques, may also be used. (Id.)

[0140] Respiratory function can be performed using a portable spirometry unit used to obtain baseline forced vital capacity (FVC) to predict disease progression or improvement. Additionally, maximum respiratory pressure, nasal inspiratory pressure (SNIP) and supping FVC may be measured and used to monitor disease progression or improvement. (Id.)

[0141] Loss of lower motor neurons is another advantage that can be utilized to monitor disease progression or improvement in ALS. Neurophysiological indices may be determined by measuring the compound muscle action potential (CMAP) in motor nerve conduction studies, parameters of which include CMAP magnitude and F-wave frequency. (Id. and de Carvalho, M., et al., Nerve conduction studies in amyotrophic lateral sclerosis. Muscle Nerve, 23:344-352, (2000)). Lower motor neuron unit number (MUNE) may be estimated similarly. MUNE estimates the number of remaining motor axons supplying the muscle to the maximum CMAP response through estimation of the contribution of individual motor units and is used to determine disease progression or improvement (Simon, et al., supra). Additional techniques to determine LMN loss include examining nerve excitability, electrical impedance myography, and using muscle ultrasound to detect changes in muscle thickness. (Id.)

[0142] Dysfunction of upper motor neurons is another advantage that can be used to monitor disease progression or improvement in ALS. Techniques for determining dysfunction include MRI or PET scans of the brain and spinal cord, transcranial magnetic stimulation, and measuring biomarker levels in cerebrospinal fluid (CSF).

[0143] (f) Glaucoma In addition to monitoring the improvement of cognition-related symptoms, the progression or improvement of glaucoma-related neurodegeneration can be monitored using techniques well known to those skilled in the art.By way of example and not limitation, monitoring can be performed through techniques such as measuring intraocular pressure, evaluating the optic disc or optic nerve head for damage, visual field testing for peripheral visual field loss, and imaging the optic disc and retina for tomographic analysis.

[0144] (g) Progressive supranuclear palsy (PSP) In addition to monitoring the improvement of cognition-related symptoms, the progress or improvement of neurodegeneration associated with progressive supranuclear palsy (PSP) can be monitored using techniques well known to those skilled in the art.By way of example and not limitation, monitoring can be performed through techniques such as functional assessment (activities of daily living or ADL), motor assessment, psychiatric symptom assessment, and volumetric and functional magnetic resonance imaging (MRI).

[0145] The level of functioning of a subject in terms of independence, partial dependence on others, or complete dependence can be useful for determining progression or improvement in the disease. (See Duff, K, et al., Functional impairment in progressive supranuclear palsy, Neurology, 80:380-84, (2013)). The Progressive Supranuclear Palsy Rating Scale (PSPRS) is a rating scale that includes 28 metrics in six categories: daily activities (by medical history), behavior, bulbar, eye movement, limb movement, and gait / midline. Performance is a score ranging from 0 to 100. Six items are scored 0 to 2, and 22 items are scored 0 to 4 for a possible total of 100. PSPRS scores are practical measures and are stringent predictors of patient survival. They are also sensitive to disease progression and are useful for monitoring disease progression or improvement. (Golbe LI, et al., A clinical rating scale for progressive supranuclear palsy, Brain, 130:1552-65, (2007)).

[0146] Functional activity can also be quantified in PSP subjects using the ADL category derived from the UPDRS (Unified Parkinson's Disease Rating Scale). (Duff, et al., supra). Similarly, the Schwab and England Activities of Daily Living score (SE-ADL) can be used to assess independence. (Id.). Additionally, the UPDRS motor function category is useful as a reliable criterion for assessing disease progression in PSP patients. The motor category may contain, for example, 27 different assessment criteria for quantifying motor function in PSP patients. Examples of these include resting tremor, stiffness, finger tapping, posture, and gait. Subject disease progression or improvement may be assessed by performing a baseline neuropsychological assessment completed by trained medical personnel using the Neuropsychiatric Inventory (NPI) to determine the frequency and severity of behavioral abnormalities (e.g., delusions, hallucinations, agitation, depression, anxiety, intense elation, anesthesia, disinhibition, irritability, and abnormal motor behavior). (Id.)

[0147] Functional MRI (fMRI) can be employed to monitor disease progression or improvement as well. fMRI is a technique that uses MRI to measure changes in brain activity in specific regions of the brain, usually based on blood flow in those regions. Blood flow is considered to correlate with activation of brain regions. Patients with neurodegenerative diseases such as PSP may be subjected to physical or intelligence tests before or while being scanned in an MRI scanner. By way of example and not limitation, the test can be the well-established force control paradigm in which the hand most affected in PSP and the patient when asked to exert force at maximal voluntary contraction (MVC) are measured by fMRI immediately after the test is performed. (Burciu, RG, et al., Distinct patterns of brain activity in progressive supranuclear palsy and Parkinson's disease, Mov. Disord. 30(9):1248-58(2015)).

[0148] Volumetric MRI is a technique in which an MRI scanner measures volumetric differences in regional brain volumes. This may be done, for example, by contrasting various disorders or by measuring the volumetric differences in brain regions in patients over time. Volumetric MRI may be employed to determine disease progression or improvement in neurodegenerative diseases such as PSP. The technique is well known to those skilled in the art. (Messina, D, et al., Patterns of brain atrophy in Parkinson's disease, progressive supranuclear palsy and multiple system atrophy, Parkinsonism and Related Disorders, 17(3):172-76(2011)). Examples of brain regions that may be measured include, but are not limited to, intracranial volume, cerebral cortex, cerebellar cortex, thalamus, caudate lobe, pallidum, globus pallidus, hippocampus, amygdala, lateral ventricle, third ventricle, fourth ventricle, and brain stem.

[0149] (h) Neurogenesis The present invention also contemplates treating or improving neurogenesis in subjects with reduced or impaired neurogenesis, which may manifest itself, for example, through reduced cognitive or motor function, or through association with neuroinflammation.Embodiments of the present invention include, by way of example and not limitation, administering blood plasma, plasma fraction or PPF to subjects with reduced or impaired neurogenesis using a pulsed dosing regimen.

[0150] The present invention also contemplates determining the level of neurogenesis before, during and / or after administration of blood plasma, plasma fraction or PPF. Non-invasive techniques for evaluating neurogenesis have been reported. (Tamura, Y. et al., J.Neurosci. (2016), 36(31): 8123-31). Positron emission tomography (PET) used with the tracer [18F]FLT in combination with the BBB transporter inhibitor probenecid allows accumulation of the tracer in neurogenesis areas in the brain. Such imaging allows evaluation of neurogenesis in patients being treated for neurodegenerative diseases.

[0151] (i) Neuroinflammation The present invention also contemplates treating or ameliorating neuroinflammation in subjects with increased neuroinflammation, which may manifest itself, for example, through reduced cognitive or motor function, or through reduced neurogenesis or association with neuroinflammation.Embodiments of the present invention include, by way of example and not limitation, administering blood plasma, plasma fractions or PPF to subjects with neuroinflammation using a pulsatile dosing regimen.

[0152] Embodiments of the invention also contemplate determining the level of neurogenesis before, during and / or after administration of blood plasma, plasma fractions or PPF. For example, 11Non-invasive techniques for assessing neuroinflammation have been reported, such as TSPO positron emission tomography (TSPOPET) using C-PK11195 and other such tracers (see Vivash, L, et al., J. Nucl. Med. 2016, 57:165-68; and Janssen, B, et al., Biochim. et Biophys. Acta, 2016, 425-41, which are incorporated herein by reference). Invasive techniques for assessing neuroinflammation include collecting cerebrospinal fluid and detecting the expression levels of neuroinflammatory markers or factors, such as, but not limited to, prostaglandin E2, cyclooxygenase-2, TNF alpha, IL-6, IFN gamma, IL-10, eotaxin, beta-2 microglobulin, VEGF, glial cell line-derived neurotrophic factor, chitotriosidase-1, MMP-9, CXC motif chemokine 13, terminal complement complex, chitinase-3-like protein 1, and osteopontin. (See Vinther-Jensen, T, et al., Neruol Neurimmunol Neuroinflamm, 2016, 3(6):e287; and Mishra, et al., J.Neuroinflamm., 2017, 14:251, which are incorporated herein by reference).

[0153] 14. Combination of stem cell and pulsed drug therapy An embodiment of the invention includes treating a subject diagnosed with cognitive impairment, impaired motor function, neuroinflammation, or reduced neurogenesis by administering an effective amount of blood plasma or a plasma fraction to a subject who is undergoing, will undergo, or has undergone stem cell therapy. Another embodiment of the invention includes administering an effective amount of blood plasma or a plasma fraction to a subject who is undergoing, will undergo, or has undergone stem cell therapy, and the stem cells used in the therapy can be embryonic stem cells, non-embryonic stem cells, induced pluripotent stem cells (iPSCs), umbilical cord blood stem cells, amniotic fluid stem cells, etc.

[0154] Stem cell therapies and techniques for performing such therapies are known to those of skill in the art (Andres, RH, et al., Brain, 2011, 134; 1777-89; Daadi, MM, et al., Cell Transplant, 2013, 22(5):881-92; Horie, N, et al., Stem Cells, 2011, 29(2):doi:10.1002 / stem.584; Thomsen, GM, et al., Stem Cells, 2018, doi:10.1002 / stem.2825; U.S. Patent Application Nos. 09 / 973,198; 12 / 258,210; 12 / 596,884; and 13 / 290,439, all of which are incorporated herein by reference). Another embodiment of the invention includes treating a subject diagnosed with traumatic spinal cord injury, stroke, retinal disease, Huntington's disease, Parkinson's disease, Alzheimer's disease, hearing loss, heart disease, rheumatoid arthritis, severe burns or in need of a bone marrow transplant and who is undergoing, will undergo, or has undergone stem cell therapy with an effective amount of blood plasma or plasma fraction.

[0155] 15. Method of screening compositions Also provided are methods of screening compositions for activity in treating cognitive or motor disorders, reducing neuroinflammation, or increasing neurogenesis. Such methods are contemplated by the present invention, including those described in the experimental examples below. Compositions that may be screened by embodiments of the present invention include biological compositions (e.g., proteins, protein combinations, antibodies, small molecule antagonists), plasma fractions, or other blood compositions. Results from the methods of screening compositions include, but are not limited to, inflammation / neuroinflammation marker results in the hippocampus (e.g., dentate gyrus) or other CNS regions, cell proliferation results in the hippocampus or other CNS regions, cell survival in the hippocampus or other CNS regions, cell fate (e.g., astrocytes, new neurons) of proliferating neural progenitor cells (NPCs) in the hippocampus or other CNS regions, and neurogenesis in the hippocampus or other CNS regions.

[0156] Another embodiment of the method of screening a composition for activity in treating cognitive or motor disorders, reducing neuroinflammation, or increasing neurogenesis includes determining the acute effect of the composition on hippocampal inflammation and proliferation in a rodent or another animal model, comprising daily administration of BrdU for 5-7 consecutive days with concomitant daily administration of the composition to be screened or a control (administered in a pulsatile manner) for 5-7 consecutive days. After the conclusion of the pulsatile dosing of the composition to be screened, up to 10 days (i.e., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 days), the number of cells in the dentate gyrus is determined by BrdU staining, and the percent of the area showing CD68 staining (indicative of inflammation) is determined.

[0157] Another embodiment of the method of screening a composition for activity in treating cognitive or motor disorders, reducing neuroinflammation, or increasing neurogenesis includes administering BrdU for 5 consecutive days (once a day) prior to commencing a 5-7 day pulsed dosing regimen of the composition or control to be screened in a rodent or another animal model, and 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks later, hippocampal cell survival is determined as the number of cells in the dentate gyrus that stain with BrdU, neurogenesis is determined as the number of cells in the dentate gyrus that stain with doublecortin (DCX), and the cell fate of neural progenitor cells to become either astrocytes (age-associated) or neurons (age-unassociated) is determined by co-localization of BrdU with GFAP or with NeuN, respectively.

[0158] Another embodiment of a method for screening a composition for activity in treating cognitive or motor disorders, reducing neuroinflammation, or increasing neurogenesis includes administering BrdU simultaneously (and daily) with the composition to be screened or a control for 5-7 days, followed by determining the extent of neurogenesis by DCX staining in the hippocampus, or the cell fate of proliferating NPCs as described above.

[0159] Another embodiment of a method for screening a composition for activity in treating cognitive or motor disorders, reducing neuroinflammation, or increasing neurogenesis includes administering a pulsed dosing regimen of the composition to be screened or a control and determining improvement in cognitive or motor function in a rodent or another animal model as described in the Examples below.

[0160] 16. Reagents, Equipment and Kits Also provided are reagents, devices and kits thereof for practicing one or more of the methods described above. The subject reagents, devices and kits thereof may vary widely.

[0161] Reagents and devices of interest include those mentioned above with respect to the methods of preparing blood products, including plasma, for infusion into a subject in need thereof, e.g., anticoagulants, cryopreservatives, buffers, isotonic solutions, etc.

[0162] The kit may also include blood collection bags, tubing, needles, centrifuge tubes, etc. In yet other embodiments, the kit as described herein includes two or more containers of blood plasma products, such as, for example, plasma protein fractions, e.g., three or more, four or more, five or more containers, including six or more of blood plasma products. In some cases, the number of different containers of blood plasma products in the kit may be 9 or more, 12 or more, 15 or more, 18 or more, 21 or more, 24 or more, 30 or more, including 36 or more, e.g., 48 or more. Each container may be associated with identifying information that includes various data about the blood plasma product contained therein, which may include one or more of the age of the donor of the blood plasma product, processing details regarding the blood plasma product, e.g., whether the plasma product has been processed to remove proteins above an average molecular weight (e.g., as described above), blood type details, etc. In some cases, each container in the kit includes identifying information for the blood plasma contained therein, including information about the age of the donor of the blood plasma product, for example, the identifying information provides identifying age-related data of the donor of the blood plasma product (such identifying information may be the age of the donor at the time of collection). In some cases, each container in the kit contains a blood plasma product derived from a donor of substantially the same age, i.e., the containers contain products derived from donors that are substantially the same age, although not all of the containers are the same. By substantially the same age, it is meant that the various donors from whom the blood plasma products of the kit are obtained differ in age, in some cases, by 5 years or less, e.g. 4 years or less, e.g. 2 years or less, e.g. 3 years or less, including 1 year or less, e.g. 9 months or less, 6 months or less, 3 months or less, including 1 month or less. The identifying information can be present on any convenient component of the container, such as, for example, a label, an RFID chip, etc. The identifying information can be human readable or computer readable as desired. The container can have any convenient shape. While the volume of the container may vary, in some cases the volume ranges from 10 mL to 5000 mL, such as from 25 mL to 2500 mL, such as from 50 mL to 1000 mL, including from 100 mL to 500 mL.The containers may be rigid or flexible and may be manufactured from any convenient material, e.g., polymeric materials, including medical grade plastic materials. In some cases, the containers have the shape of a bag or pouch. In addition to the containers, such kits may further include administration equipment, e.g., as described above. The components of such kits may be provided in suitable packaging, e.g., a box or similar structure configured to hold the containers and the other kit components.

[0163] In addition to the above components, the subject kits further include instructions for practicing the subject methods. These instructions may be present in the subject kits in a variety of forms, one or more of which may be present in the kit. One form in which these instructions may be present is as information printed on a suitable medium or substrate, such as one or more sheets of paper on which the information is printed, such as on the kit's packaging, on a package insert, etc. Yet another means is a computer readable medium on which the information is recorded, such as a disk, CD, portable flash drive, etc. Yet another means that may be present is a website address that may be used via the internet to access the information at the transferred site. Any convenient means may be present in the kit.

[0164] 17. Exercise load Exercise loads can be characterized as aerobic or non-aerobic activities and can include high calorie burning activities and moderate calorie burning activities. Exercise loads can include strength training (e.g., weight training or isometric loading). Exercise loads can also include, for example, running, biking, walking, dancing, marching, swimming, yoga, tai chi, balance loads, leg bands, skipping rope, surfing, rowing, arm or leg rotations and flexions, gardening, cleaning, vigorous sports such as bowling, aerobics, pilates, and martial arts.

[0165] The exercise stress plan may include performing a single exercise stress at a specific frequency or performing a combination of exercise stresses at a specific frequency. The frequency may be 1, 2, 3, 4, 5, 6 or 7 times per week. The frequency may vary from week to week. The exercise stress plan may be at the same level of intensity and / or frequency as the subject performed before administration of the composition of the present invention. The exercise stress plan may be at a higher level of intensity and / or frequency compared to the level performed by the subject before administration of the composition of the present invention. The exercise stress plan may be suggested or prescribed by a medical or fitness professional, or the exercise stress plan may be initiated by the subject himself or herself. EXAMPLES

[0166] 18. Experimental Examples (a) Example 1 Clarified young human plasma (young plasma) or commercially available PPF ("PPF1") was administered to aged immunodeficient mice (NOD.Cg-Prkdscid Il2rgtm 1 Wj1 / SzJ, "NSG" strain). PPF1 is PPF with approximately 88% normal human albumin (based on total protein), 12% alpha and beta globulins, and less than 1% gamma globulins, as determined by electrophoresis. Except where noted, PPF1 is administered in vivo in the examples herein using a 5% solution (w / v, 50 g / L). All mice were homogenized across treatment groups according to four different criteria: nesting score in their own cage, initial body weight, distance traveled in the open field, and % time in the center of the open field. Following group assignment, mice were injected intraperitoneally (IP) with BrdU (5-bromo-2'-deoxyuridine) formulated in PBS (phosphate buffered saline) at a final concentration of 10 mg / mL administered at 150 mg / kg for 5 days. Following this, mice were injected intravenously (IV) with 150 μL of PPF1 three times per week for 4 weeks. Behavioral testing occurred during weeks 5 and 6, when mice received two injections per week to avoid injections on parallel test days. Mice were euthanized 24 hours after the last IV injection for a total of 16 injections over the 6-week period. Two additional cohorts of mice were injected intravenously (IV) with 150 μL of PPF1 or saline for 7 consecutive days (pulsed). Behavioral testing occurred during weeks 5 and 6, with the same number of injections per week, 3 per group.

[0167] Behavioral tests were analyzed using CleverSys software (Reston, VA). CleverSys TopSCan V3.0 was used to track mouse behavior in the no-maze, Barnes maze, open field, and Y-maze. The Barnes maze was constructed by CleverSys. The grip strength meter was designed and manufactured by Columbus Instruments (Columbus, OH). The Y-maze and open field chambers were constructed according to the specifications of San Diego Instruments (San Diego, CA). Histological analysis of hippocampal slices was performed with a Leica (Buffalo Grove, IL) imaging microscope model DM5500B equipped with a DCF7000T brightfield / fluorescence microscope camera.

[0168] Behavioral testing FIG. 1A shows that the group pulse-treated with PPF1 tended to increase the distance traveled in the open field test compared to both the saline control group and the group treated with PPF1 three times a week. This result indicates a trend toward increased mobility in the pulse-treated group. FIG. 1B shows that the group pulse-treated with PPF1 tended to increase the proportion of time spent in the center of the open field compared to both the saline control group and the group treated with PPF1 three times a week. This result indicates a trend toward decreased anxiety in the pulse-treated group.

[0169] body weight Figure 2 shows the effect of PPF1 on body weight. Both PPF1 treatment groups (pulse dosing or three times weekly) showed no adverse effects from the injections.

[0170] histology Figure 3 shows the number of DCX positively labeled cells in the granular layer of the dentate gyrus. There was a significant increase in neurogenesis between the 3x / week treatment group and the pulsatile PPF1 treatment group compared to the saline group. All data shown are means ± sem. **p<0.01. One-way ANOVA with Dunnett's multiple comparison post-hoc analysis (n: saline=8, PPF1 pulsed=10, PPF1 3x / week=10). Figure 4 shows the number of BrdU positively labeled cells in the granular layer of the dentate gyrus of three differently treated groups of mice. There was a significant increase in cell survival between the 3x / weekly treated group and the pulsed PPF1 treated group compared to the saline group. All data shown are means ± sem. **** p<0.0001, * p<0.05. One-way ANOVA with Dunnett's multiple comparison post-hoc analysis (n: saline=8, PPF1 pulse=10, PPF1 3× / week=10).

[0171] Analysis of hippocampal slices was performed on a Leica (Buffalo Grove, IL) imaging microscope model DM5500B equipped with a DCF7000T brightfield / fluorescence microscope camera. Ki67 staining was Abcam (ab15580) at 1:500 and the secondary antibody was goat anti-rabbit (Alex Fluor555) (ab150090) at 1:300.

[0172] (b) Example 2 Clarified young human plasma (YP), aged human plasma (OP) or commercial PPF ("PPF1") was administered to aged immunodeficient mice (NOD.Cg-Prkdscid Il2rgtm 1 Wj1 / SzJ, "NSG" strain). All mice were homogenized across treatment groups according to four different criteria: scoring for nesting in their cage, initial body weight, distance traveled in the open field, and % of time spent in the center of the open field. Following group determination, mice were injected intraperitoneally (IP) with BrdU formulated in PBS (phosphate buffered saline) at a final concentration of 10 mg / mL administered at 150 mg / kg for 5 days. Following this, mice were injected intravenously (IV) with 150 μL of either clarified young human plasma or PPF1 (1) three times per week for 6 weeks ("3x / week"), (2) three times per week for 1 week only ("3x"), or (3) for 7 days for 1 week. Additional groups of mice received saline pulses for 7 days. Six weeks after the start of dosing with young or aged plasma, PPF1, or vehicle, all mice were sacrificed.

[0173] histology FIG. 5 shows the number of DCX positively labeled cells in the granular layer of the dentate gyrus in nine separate treatment groups of mice treated with either young human plasma (YP), aged human plasma (OP), PPF1 or saline. Mice treated with PPF1 showed increased neurogenesis compared to other groups, both pulsed or treated three times a week. All data shown are means ± sem. * p<0.05. ANOVA with Dunnett's post hoc analysis. PPF1 treatment (pulsed or 3x / week) and saline treatment (n: saline=4, PPF1 pulsed=5, PPF1 3x / week=5, PPF1 3x=4, YP pulsed=6, YP3x / week=6, YP3x=4, AP pulsed=6, AP3x=6).

[0174] FIG. 6 shows the number of BrdU positively labeled cells within the granular layer of the dentate gyrus in nine separate treatment groups of mice treated with either young human plasma (YP), aged human plasma (OP), PPF1 or saline. Mice treated with PPF1 showed a significant increase in cell survival compared to the other groups, with pulsed PPF1-treated mice showing a greater significant difference than mice treated with 3x weekly PPF1. All data shown are mean ± sem. ** p<0.01, * p<0.05. ANOVA with Dunnett's post hoc analysis. PPF1 treatment (pulsed or 3x / week) and saline treatment (n: saline=4, PPF1 pulsed=5, PPF1 3x / week=5, PPF1 3x=4, YP pulsed=6, YP3x / week=6, YP3x=4, AP pulsed=6, AP3x=6).

[0175] (c) Example 3 Clarified young human plasma (YP), aged human plasma (OP) or commercial PPF ("PPF1") was administered to aged immunodeficient mice (NOD.Cg-Prkdscid Il2rgtm 1 Wj1 / SzJ, "NSG" strain). Mice were treated intravenously (IV) daily for 7 days for a weekly dosing regimen.

[0176] All mice were homogenized across treatment groups according to four different criteria: scoring for nesting in their own cage, initial body weight, distance traveled in the open field, and % time spent in the center of the open field. Following group determination, mice were injected intraperitoneally (IP) with BrdU formulated in PBS (phosphate buffered saline) at a final concentration of 10 mg / mL administered at 150 mg / kg for 5 days. All mice were injected intravenously (IV) with 150 μL of young or old plasma, PPF1, or saline for 7 consecutive days (referred to as pulse dosing). Three weeks after pulse dosing was completed, mice were injected intraperitoneally (IP) with EdU (5-ethynyl-2'-deoxyuridine) formulated in PBS (phosphate buffered saline) at a final concentration of 10 mg / mL administered at 30 mg / kg for 5 days. At week 8 (6 weeks following the end of pulse dosing), the Barnes maze was performed.

[0177] Behavioral tests were analyzed using CleverSys software (Reston, VA). CleverSys TopSCan V3.0 was used to track mouse behavior in the Barnes maze. Analysis of hippocampal slices was performed on a Leica (Buffalo Grove, IL) imaging microscope model DM5500B equipped with a DCF7000T brightfield / fluorescence microscope camera.

[0178] Behavioral testing Figure 7 shows the time taken to find the target hole (latency) per trial per day for each treatment group when tested in the Barnes maze. Mice treated with pulsed administration of PPF1 showed a significant decrease in latency to trial for several individual test segments, indicating improved cognitive performance. * p<0.05. Mean ± sem, independent t-test (n: saline=13, PPF1=13, AP=14, YP=14).

[0179] histology FIG. 8 shows the number of DCX positively labeled cells in the granular layer of the dentate gyrus in four separate treatment groups treated with either young human plasma (YP), aged human plasma (OP), PPF1 or saline. There was a significant increase in neurogenesis with pulsed PPF1 and pulsed young human plasma compared to saline treatment. All data shown are mean±sem. **** p<0.0001, ** p<0.01. One-way ANOVA with Dunnett's multiple comparison post-hoc analysis. (n: Saline=14, PPF1=14, AP=14, YP=15).

[0180] Figure 9 shows the number of BrdU-labeled cells in the granular layer of the dentate gyrus of mice treated with either young human plasma (YP), aged human plasma (OP), PPF1 or saline. There was a significant increase in cell survival with pulsed PPF1 and pulsed YP compared to saline treatment. All data shown are means ± sem. ****p<0.0001. Mean ± SEM, one-way ANOVA with Dunnett's multiple comparison post-hoc analysis. (n: Saline=14, PPF1=14, AP=14, YP=15).

[0181] (d) Example 4 Commercially available PPF ("PPF1") was administered to aged immunodeficient mice (NOD.Cg-Prkdscid Il2rgtm 1 Wj1 / SzJ, "NSG" strain). Twelve-month-old mice were treated with intravenous tail vein (IV) dosing daily for 7 days on a one-week dosing regimen. After treatment, mice were left in their own cage environment for 4.5 weeks prior to behavioral testing. All injections and behavioral testing were performed over the course of 7 weeks for each cohort, for a total period of 9 weeks. All mice received BrdU IP for 5 days prior to the first dose. Mice were sacrificed one day after the conclusion of the last behavioral test.

[0182] Behavioral tests were analyzed using CleverSys software (Reston, VA). CleverSys TopSCan V3.0 was used to track mouse behavior in the Y-maze.

[0183] Behavioral testing FIG. 10 shows the percentage of total entries into familiar or novel lanes out of total entries into each lane by treatment group in the Y-maze test. Twelve-month-old mice were pulse-treated with saline, PPF1, or 5× concentration of PPF1. Both pulse-treated PPF1 and PPF1(×5) mice showed a significant increase in novel lane entries compared to the amount of novel lane entries by saline-treated mice, indicating improvement in cognition. All data shown are mean±sem. * p<0.05. Paired t-test.

[0184] Figure 11 shows the ratio of entries into the novel lane to the familiar lane in the Y-maze for each treatment group. Twelve-month-old mice were pulse-treated with saline, PPF1, or a 5x concentration of PPF1. Both pulse-treated PPF1 and PPF1 (x5) mice showed a trend toward increased entries into the novel lane compared to saline-treated mice. All data shown are mean ± sem.

[0185] histology Figure 12 shows the number of BrdU positively labeled cells in all hippocampal slices. Mice treated with pulsed doses of PPF1 showed a trend towards increased cell survival compared to saline and PPF1 (x5) treated mice. All data shown are mean ± sem.

[0186] Figure 13 shows the number of DCX positively labeled cells in all hippocampal sections. Mice pulsed with PPF1 and PPF1(x5) showed a trend towards increased neurogenesis compared to saline-treated mice. All data shown are mean ± sem.

[0187] (e) Example 5 Commercially available PPF ("PPF1") was administered to aged (10.5 months old) immunodeficient mice (NOD.Cg-Prkdscid Il2rgtm 1 Wj1 / SzJ, "NSG" strain). All mice were homogenized across treatment groups according to four different criteria: scoring for nesting in their cage, initial body weight, distance traveled in the open field, and % of time spent in the center of the open field. Following group determination, mice were injected intraperitoneally (IP) with BrdU formulated in PBS (phosphate buffered saline) at a final concentration of 10 mg / mL administered at 150 mg / kg for 5 days. Following this, mice were injected intravenously (IV) with PPF1 for either (1) 5 consecutive days [PPF1-5d], (2) 7 consecutive days [PPF1-7d], (3) 5 consecutive days and an additional 5 consecutive days of enhanced (B) dosing occurring 6 weeks after completion of the first dosing [PPF1-5d-B], or (4) 7 consecutive days and an additional 7 consecutive days of enhanced (B) dosing occurring 6 weeks after completion of the first dosing [PPF1-7d-B]. Additional groups were injected with saline for 7 consecutive days and an additional 7 consecutive days of enhanced (B) dosing occurring 6 weeks after completion of the first dosing [SAL-7d-B]. After 5 weeks of pulse dosing, mice were injected IP for 5 days with EdU (5-ethynyl-2'-deoxyuridine) formulated in PBS at a final concentration of 10 mg / mL dosed at 30 mg / kg. All mice were sacrificed 12 weeks after completion of pulse dosing with PPF1 or vehicle.

[0188] Analysis of hippocampal slices was performed with a Leica (Buffalo Grove, IL) imaging microscope model DM5500B equipped with a DCF7000T brightfield / fluorescence microscope camera. Figure 14 shows the number of DCX positively labeled cells in the granular layer of the dentate gyrus in PPF1 and saline treated mice. These results show a significant improvement in the group treated for 5 consecutive days followed by enhanced dosing, which is comparable to the group treated for 7 consecutive days. All data shown are mean ± sem. PPF1-7d, PPF1-5d-B vs saline *p<0.05. ANOVA with Dunnett's post hoc analysis. (n: Saline=5, PPF1-5d=8, PPF1-7d=7, PPF1-5d-B=8, PPF1-7d-B=7).

[0189] FIG. 15 shows the number of BrdU positively labeled cells in the granular layer of the dentate gyrus in mice treated with PPF1 and saline. These results indicate that stimulation of five consecutive days of treatment followed by the boosted dose results in a substantial increase in proliferating cells compared to five consecutive days or seven consecutive days without the boosted dose. Furthermore, the boosted dose treatment significantly increases cell survival overall. All data shown are means ± sem. PPF1-5d-B, PPF1-7d-B vs. saline *** p<0.001, * p<0.05. ANOVA with Dunnett's post hoc analysis. PPF1-5d vs. PPF1-5d-B, +p<0.05, unpaired t-test. (n: saline=7, PPF1-5d=8, PPF1-7d=7, PPF1-5d-B=8, PPF1-7d-B=7).

[0190] Figure 16 shows the number of EdU positively labeled cells in the granular layer of the dentate gyrus in mice treated with young plasma, PPF1 and saline. These results indicate that the effect observed with the boosted dose is not due to an increase in the total number of proliferating cells present, but due to enhanced survival mechanisms elicited by the boosted dose. All data shown are mean ± sem. (n: saline = 4, PPF1-5d = 7, PPF1-7d = 6, PPF1-5d-B = 7, PPF1-7d-B = 6).

[0191] (f) Example 6 Commercially available PPF ("PPF1") was administered to adult (3 and 6 months old) immunodeficient mice (NOD.Cg-Prkdscid Il2rgtm 1 Wj1 / SzJ, "NSG" strain). All mice were homogenized across treatment groups according to four different criteria: scoring for nesting in their cage, initial body weight, distance traveled in the open field, and % time spent in the center of the open field. Following group determination, mice were injected intraperitoneally (IP) with BrdU formulated in PBS (phosphate buffered saline) at a final concentration of 10 mg / mL administered at 150 mg / kg for 5 days. Following this, mice were injected intravenously (IV) with saline or PPF1 for 7 consecutive days (pulse dosing). A subset of mice from both saline and PPF1 treatments were provided with a running wheel in their cage. Mice were sacrificed 3, 10, or 42 days after completion of pulse dosing.

[0192] Figure 17 shows the number of DCX positively labeled cells in the granular layer of the dentate gyrus in 3-month-old NSG animals treated with PPF1 or saline treatment with or without a running wheel. All data shown are mean ± sem. Saline vs. Wheel + PPF1 42 days, Wheel 42 days, **** p<0.0001, * p<0.05. ANOVA with Dunnett's post hoc analysis. Wheel running vs. 42 days after PPF1. *** p<0.0001, unpaired t-test. (n: saline 3 days=8, PPF1 3 days=8, PPF1 10 days=7, vehicle 42 days=8, PPF1 42 days=8, wheel 42 days=8, wheel+PPF1 42 days=8). Figure 17 also shows the number of DCX positively labeled cells in the granular layer of the dentate gyrus in 6-month-old NSG animals treated with PPF1 or saline treatment with or without a wheel. All data shown are mean±sem. Wheel+PPF1 42 days vs. saline 42 days, wheel 42 days, **** p<0.0001, ** p<0.01. ANOVA with Dunnett's post hoc analysis. Wheel running vs. 42 days after PPF1. ***p<0.0001, unpaired t-test. After 42 days of PPF1 vs. after 42 days of saline. * p<0.05, unpaired t-test. (n: 3 days after saline = 7, 3 days after PPF1 = 8, 10 days after PPF1 = 6, 42 days after saline = 8, 42 days after PPF1 = 6, 42 days after running wheel = 8, 42 days after running wheel + PPF1 = 9).

[0193] Figure 18 shows the number of Ki67 positively labeled cells in the granular layer of the dentate gyrus in 3-month-old NSG animals treated with PPF1 or saline treatment with or without a running wheel. All data shown are mean ± sem. After 42 days of running wheel + PPF1 vs. saline, *** p<0.001, ANOVA with Dunnett's post hoc analysis. (n: 3 days saline=6, 3 days PPF1=6, 10 days PPF1=7, 42 days saline=8, 42 days PPF1=8, 42 days wheel=8, 42 days wheel + PPF1=8).

[0194] Figure 18 also shows the number of Ki67 positively labeled cells in the granular layer of the dentate gyrus in 6-month-old NSG animals treated with PPF1 or saline treatment with or without a running wheel. All data shown are mean ± sem. Saline vs. Wheel + PPF1 42 days, Wheel 42 days, *** p<0.001, * p<0.05. ANOVA with Dunnett's post hoc analysis. (n: 3 days saline=7, 3 days PPF1=7, 10 days PPF1=8, 42 days saline=8, 42 days PPF1=7, 42 days wheel=7, 42 days wheel+PPF1=9).

[0195] Figure 19 shows the number of BrdU positively labeled cells in the granular layer of the dentate gyrus in 3 and 6 month old NSG animals treated with PPF1 or saline treatment with or without running wheels. All data shown are mean ± sem. After 42 days of running wheels + PPF1 vs vehicle, *** p<0.001, ANOVA with Dunnett's post-hoc analysis.**** p<0.0001, *** p<0.001, ** p<0.01, * p < 0.05, ANOVA with Dunnett's post hoc analysis).

[0196] These data show that there is a significant increase in neurogenesis in PPF1 and running wheel compared to vehicle after 6 weeks of dosing in 3 month old NSG mice. Additionally, there is a significant increase in neurogenesis in PPF1 and running wheel compared to wheel alone after 6 weeks of dosing in 3 month old NSG mice. There is also a significant increase in neurogenesis in PPF1 and running wheel compared to vehicle after 6 weeks of dosing in 6 month old NSG mice. These results also show a significant increase in neurogenesis in PPF1 and running wheel compared to wheel alone after 6 weeks of dosing in 6 month old NSG mice. Additionally, there is a significant increase in progenitor cell proliferation in PPF1 and running wheel compared to vehicle after 6 weeks of dosing in both 3 month old and 6 month old NSG mice.

[0197] These findings in adult NSG mice at 6 months of age indicate a potential synergistic effect of exercise and PPF1 administration that resulted in a significant enhancement of neurogenesis compared to either exercise or PPF1 treatment separately, supporting the potential utility of PPF1 treatment in conjunction with an exercise regimen in a clinical setting. Furthermore, these data reveal a significant capacity for neurogenesis in the brain that may be accessed via multiple independent or overlapping mechanisms.

[0198] (g) Example 7 PPF1 or saline control was administered to two treatment groups of 11-month-old immunodeficient mice (NOD.Cg-Prkdscid Il2rgtm 1 Wj1 / SzJ, "NSG" strain). All mice received IV injections of 150 μL of PPF1 or saline per dose for seven consecutive days. Running wheels (MedAssociates) were placed in the cages of mice, named runners (n=8, n=8 for PPF1 and saline) beginning in the seventh week of the study. The number of wheel revolutions was recorded during the day and night for five consecutive days.

[0199] FIG. 20 shows the number of wheel revolutions in a given period with the shaded area indicating the dark phase. The statistical significance of total running for both treated and untreated groups in the light-dark cycle was assessed using an unpaired t-test. Rhythm expression profiles were extracted and characterized using regional analysis of time and frequency for the 13 time-point series separately for each mouse from treated and untreated groups with five 13 time-point series per mouse. Duration, phase and magnitude were parameters defined for each rhythm and compared between the two groups using an unpaired two-tailed t-test. Mice treated with PPF1 ran significantly more than untreated mice, an indication of improved locomotor activity. Mice were subjected to the hot plate test to check for normal pain sensation in the paws. The loss of sensation should have had an effect prior to the behavioral readout. The hot plate test led to a mild increase in activity after return to the running wheel cage environment as evidenced by the spike in wheel revolutions shown in the boxed section of FIG. 20.

[0200] (h) Example 8 Recombinant human albumin ("rhAlbumin, Albumedix, Ltd, Nottingham, UK), clarified young human plasma ("YP") or saline control were administered to 10.5 month old immunodeficient mice (NOD.Cg-Prkdscid Il2rgtm 1 Wj1 / SzJ, "NSG" strain). All mice received 50mg / kg BrdU IP on week 1 prior to 7 days of pulse dosing. rhAlbumin and YP were diluted to 50mg / mL in water for injection (WFI, 0.9% saline). All mice received 7 consecutive days of IV injections of 150μL rhAlbumin, YP or saline per dose. Six weeks after the final day of treatment, mice were sacrificed.

[0201] FIG. 21A shows the amount of cell survival in all three treatment groups as determined by the number of BrdU-labeled cells in the dentate gyrus ("DG"). Young plasma significantly increased cell survival compared to saline and rhalbumin, whereas rhalbumin had no significant effect on cell survival. All data shown are means±sem (paired t-test). *** p<0.001).

[0202] FIG. 21B shows the amount of DCX staining in all three treatment groups as determined by the number of DCX positive cells in the dentate gyrus ("DG"). Young plasma significantly increased neurogenesis compared to saline and rhalbumin, whereas rhalbumin was associated with decreased neurogenesis compared to saline controls. All data shown are means±sem (paired t-test). ** p<0.01, *** p<0.001).

[0203] (i) Example 9 Dissociated mixed neurons derived from mouse E16 cortex were plated and grown in 48-well multielectrode array plates (Axion Biosystems). Each well contains 16 electrodes that make physical contact with plated neurons and measure subtle changes in cell membrane properties. This setup allows the assessment of a variety of different parameters to obtain information about neuronal spiking activity and firing behavior at the single electrode level, as well as the degree of neuronal connectivity by assessing the synchrony of neuronal firing properties across multiple electrodes within a well.

[0204] Neuronal cultures were maintained in the presence of treatment conditions from day 1 onwards. Treatment conditions included Neurobasal medium containing 10% (v / v) recombinant human albumin (rhAlbumin, Albumedix, Ltd, Nottingham, UK), PPF1 or HAS1 plus B27 supplement. PBS constituted the control. Neuronal activity was measured on days 7 and 14 of culture.

[0205] FIG. 22 shows that PPF1 treatment for 7 days results in increased neuronal network activity compared to control, rhalbumin or HAS1 treatment. HAS1 is a commercially available HAS that is >95% human albumin (by total protein) in a 5% solution (w / v, 50 g / L) prepared by cold alcohol fractionation and derived from pooled human plasma from donors. Both PPF1 and HAS1 came as 5% solutions (w / v, 50 g / L) and were diluted 1:10 in Neurobasal medium + B27 supplement. The effect of PPF1 on neuronal network activity persists up to 14 days in culture, indicating that PPF1 is associated with enhanced neuronal network maturation. Data shown are mean ± sem (by unpaired t-test) * p<0.05).

[0206] (j) Example 10 Clarified aged human plasma (OP) or sterile saline was administered to 8-week-old (young) immunodeficient mice (NOD.Cg-Prkdscid Il2rgtm 1 Wj1 / SzJ, "NSG" strain). For each experiment, mice were homogenized across treatment groups by body weight. All mice were injected IP with 150 mg / kg BrdU in sterile PBS for 5 consecutive days. BrdU injections were followed by IV administration of aged plasma with different treatment regimens at 150 μL per dose. All regimens are outlined in FIG. 23.

[0207] Model 1 included two injections per week for a total of 10 injections over 5 weeks. Histological analysis was performed 48 hours after the last plasma dose. Model 2 included three injections per week for a total of 10 injections over 4 weeks with histological analysis 48 hours after the last plasma dose. In model 3, mice received daily injections for 7 consecutive days with histological analysis 48 hours after the last dose. In model 4, mice received daily injections for 7 consecutive days with analysis 21 days after the last dose. Brains of mice treated with aged plasma were analyzed for markers of endothelial inflammation, VCAM-1 in the hippocampus and the number of newborn neurons labeled by doublecortin (DCX) positive cells in the dentate gyrus. VCAM-1 was processed for immunohistochemistry on 30 μm floating sections, imaged on a Hamamatsu NanoZoomer HT (Hamamatsu) and analyzed using Image Pro Software (Media Cybernetics). DCX-positive cells in the dentate gyrus were counted live under a Leica wide-field microscope (Leica).

[0208] Analysis of percent VCAM-1 positive area in the hippocampus (Figure 24) showed that endothelial inflammation was significantly elevated 48 hours after the last plasma administration, with a trend toward increased significance with twice weekly dosing (Figure 24A), three times weekly dosing (Figure 24B) and pulse dosing (Figure 24C). 21 days after the last aged plasma administration, VCAM-1 was no longer significantly elevated (Figure 24D).

[0209] Since the effect on doublecortin could only be observed after a period of 3-4 weeks, the number of DCX positive cells was analyzed in the dentate gyrus in norms 1, 2 and 4. The analysis showed that there was no effect of aged plasma on neurogenesis with a twice weekly (Fig. 25A) or three times weekly (Fig. 25B) dosing norm, but pulsed dosing for 7 consecutive days (Fig. 25C) resulted in a significant decrease in the number of DCX positive cells. This data suggests that only pulsed dosing of aged human plasma had a significant effect on neurogenesis.

[0210] (k) Example 11 Eight-week-old NSG mice treated with aged human plasma (origin 65-68 years) for seven consecutive days were tested using a modified Barnes maze 4 weeks after the last injection of aged plasma. Figure 26 shows the time course for escape in the Barnes maze, showing the time to reach and enter the escape hole for NSG mice treated with aged plasma and saline. There was no significant difference in the time to escape between groups, but mice treated with aged plasma on day 4 performed less well than saline controls. This data indicates impaired learning and memory in a spatial memory task related to hippocampal function. All data shown are mean ± sem. Two-way ANOVA, Sidak post-hoc test.

[0211] Figure 27 shows the mean escape latencies in the last three Barnes maze trials on day 4. Mice treated with aged plasma showed a trend towards increased escape latencies, indicating impaired memory function. All data shown are mean ± sem. (paired t-test).

[0212] Figure 28 shows the difference in escape times between trial 1 and trial 3 of the Barnes maze, indicating that these trials can be used as a metric for learning within a day. Mice treated with aged plasma had significantly less difference in escape times between these trials, revealing a reduced learning ability. All data shown are means ± sem. (by unpaired t-test) * p<0.05).

[0213] Figure 29 shows the results of qPCR used to quantify the mRNA levels of various markers related to neurogenesis and synaptic function. The relative expression level of doublecortin (DCX), a marker of new neurons, was decreased in agreement with the histological analysis of the same marker. In addition, there was a trend towards decreased levels of vglut1 (vesicular glutamate transporter 1), a marker of glutamatergic synapses, synaptic markers syn1 (synapsin 1), tuj1 (beta III tubulin), and bdnf (brain-derived neurotrophic factor). These decreases indicate an overall impaired synaptic and neuronal network in the brains of mice injected with aged plasma. All data shown are mean ± sem. (Paired t-test) * p<0.05).

[0214] (l) Example 12 Young (8 weeks old) immunodeficient mice (NOD.Cg-Prkdscid Il2rgtm 1 Wj1 / SzJ, "NSG" strain) were homogenized across treatment groups by body weight. Mice were injected subcutaneously (sc) with 35 mg / kg kainic acid (Sigma) in sterile saline or saline control. Peripheral administration of kainic acid produced acute seizure activity, inflammation in the hippocampus, and in a subset of mice, neuronal loss in the CA1 region of the hippocampus. Two hours after kainic acid injection, mice were administered 150 μL of PPF1 or saline intravenously. Administration of PPF1 or saline continued daily for a total of 5 days (FIG. 30). Tissues were harvested for analysis on the sixth day. Inflammatory changes were analyzed in the CA1 region of the hippocampus after immunofluorescent staining for microglial activation (CD68) and astrocytic activation (GFAP). The sections were subjected to immunohistochemical processing on 30-μm floating sections, and then imaged using a Hamamatsu NanoZoomer HT (Hamamatsu) and analyzed using Image Pro Software (Media Cybernetics).

[0215] Analysis of the percent CD68 positive area in the CA1 region of the hippocampus shows that kainic acid administration results in an increase in CD68 immunoreactivity, suggesting microglial activation (Figure 31A). Five days of PPF1 administration results in a significant decrease in the percentage of CD68 positive area, thus resulting in a decrease in microglial activation. Similarly, analysis of the percentage of CFAP positive area (Figure 31B) shows a significant increase after kainic acid administration, which is significantly decreased after PPF1 dosing. The data suggest that PPF1 has an acute anti-inflammatory effect in the brains of mice receiving kainic acid. * p<0.05. One-way ANOVA with Dunnett's multiple comparison post-hoc analysis.

[0216] (m) Example 13 Six-month-old NSG mice were injected daily for one week (7 days) with either PPF1 or saline control IV at a dose of 150 μL (10 mg / mL). All mice were treated with 50 mg / kg BrdU once daily on the same day they received PPF1 or saline control. Mice were then split into two cohorts. The first cohort was sacrificed the day immediately following 7 days of co-treatment with BrdU and PPF1. The second cohort was sacrificed 7 days later and received daily BrdU for an additional 7 days.

[0217] FIG. 32 shows the number of stained cells in the dentate gyrus for cohorts 1 and 2 (left to right). Both cohorts showed increased cell proliferation in the dentate gyrus compared to saline controls ( *** p<0.001, independent t-test).

[0218] (n) Example 14 Three-month-old or six-month-old NSG mice were injected daily for one week (7 days) with either PPF1 or saline vehicle by IV. Mice were then sacrificed 3, 10, or 42 days after the seven-day daily dose was administered. Brains were stained with Ki67, a nuclear marker present only in proliferating cells that labels neural stem and progenitor cells in the blades of the dentate gyrus. Figure 33 shows that 10 days after the end of a continuous seven-day pulse regimen with PPF1, six-month-old mice exhibited an increase in total progenitor cells (Ki67 positive or "Ki67+") in the dentate gyrus. Figure 34 shows Ki67 staining (bright areas) in the dentate gyrus in NSG mice 10 days after the end of a continuous seven-day pulse regimen with PPF1. This indicates that one possible mechanism of action for PPF1 in increasing overall cell survival and neurogenesis 42 days after dosing cessation may be through an increase in total progenitor cells (neural stem cells).

[0219] (o) Example 15 Commercially available PPF ("PPF1") or saline control was administered to two different populations of 6- and 12-month-old immunodeficient mice (NOD.Cg-Prkdscid Il2rgtm 1 Wj1 / SzJ, "NSG" strain). All mice received 50 mg / kg BrdU the first week prior to a 7-day pulse dose of test agent. All mice received an IV injection of 150 μL of PPF1 or saline per dose. One cohort from each treatment group was used to study proliferation and was sacrificed 6 weeks after the last administered dose.

[0220] FIG. 35A shows that a cohort of 6-month-old mice treated with PPF1 showed a significant increase in the number of progenitor cells differentiating into neurons (NeuN+) compared to saline controls, and a decrease in the number of progenitor cells differentiating into astrocytes (GFAP+) compared to controls. All data shown are means ± sem. (Paired t-test) ** p<0.01).

[0221] FIG. 35B shows that a cohort of 12-month-old mice treated with PPF1 showed a significant increase in the number of progenitor cells differentiating into neurons (NeuN+) compared to saline controls, and a non-statistically significant difference in the number of progenitor cells differentiating into astrocytes (GFAP+) compared to controls. All data shown are means ± sem. (by unpaired t-test) ** p<0.01).

[0222] (p) Example 16 Clarified aged human plasma (aged plasma) or sterile saline was administered to 3-month-old mice (NOD.Cg-Prkdscid Il2rgtm 1 Wj1 / SzJ, "NSG" strain). For each experiment, mice were homogenized across treatment groups by body weight. All mice received IP injections of 150 mg / kg BrdU in sterile saline for 5 consecutive days. BrdU injections were followed by daily IV administrations of 150 μL of aged plasma or sterile saline per dose for 7 consecutive days, and were histologically analyzed 4 weeks after the last dose.

[0223] Figure 36 shows the cell fate of proliferating neural progenitor cells labeled with BrdU 4 weeks after the last injection. In mice injected with aged plasma, surviving BrdU-labeled cells differentiate significantly less into neurons than into astrocytes. This indicates that aged human 36B plasma alters the cell fate of neural progenitor cells to the astrocyte lineage in young mice (Figure 37B) and negatively affects the number of newborn neurons in the dentate gyrus (Figure 36A) (n=12 per group). All data shown are means ± sem. (by unpaired t-test) *** p<0.001, **** p<0.0001).

[0224] (q) Example 17 cortical activation Aged (18 months old) C57BL / 6 mice received IV injections of 150uL of PPF1 or 0.9% saline daily for 7 days. Two and a half hours (2.5 hours) after the last test agent administration, mice were sacrificed by transcardial perfusion with 0.9% saline followed by 4% formaldehyde under deep anesthesia with ketamine and xylazine. Brains were sectioned, postfixed, and processed by the iDisco procedure to visualize cFos-positive cells via light sheet fluorescence microscopy (LSFM) at a voxel resolution of 2x2x3 micrometers. Imaged brains were assigned as three-dimensional volumes, and activated cFos-positive cells were detected computationally. Statistical comparisons between groups were performed by negative binomial regression corrected for multiple comparisons by false discovery rate (* indicates p-value less than 0.05).

[0225] Analysis of mouse brains showed an overall increase in the number of cFos positive cells in the whole brain volume as well as the cortex and isocortex in 18-month-old mice treated with PPF1 (Figure 37). Using binomial regression corrected for multiple comparisons, the difference between these increases in the overall number of positive cFos did not reach significance. However, analysis of more defined cortical regions, such as the frontal cortex, orbital cortex, intralimbic cortex, and prelimbic cortex, showed a significant increase in the number of cFos positive cells (Figure 38), indicating elevated neuronal activity. Enhanced activity in prefrontal cortical regions correlated with improved cognitive performance, suggesting that PPF1 treatment results in improved cognition in aged C57BL / 6 mice. Similar significant increases in the number of cFos positive cells were also found in the accessory olfactory nucleus and olfactory tubercle (Figure 39). These regions are associated with processing olfactory information, and enhanced activity suggests increased olfactory function. Voxel statistic-based visualization of cFos activation in red showed increased cFos signal in the cortex of mice treated with PPF1 (FIG. 40).

[0226] (r) Example 18 Commercially available PPF ("PPF1") or saline control was administered to 22-month-old wild-type (WT) mice (C57BL / 6J, "WT", strain code 0664, Jackson Labs, Bar Harbor, ME). All mice received 50 mg / kg BrdU in the first week prior to a 7-day pulse dosing. Subsequently, all mice received IV injections of 150 μL of PPF1 or saline per dose for 7 consecutive days. Ten days after the last PPF1 or saline injection, mice were sacrificed and brains processed for histology.

[0227] Figure 41A shows the percentage of CD68 immunoreactive area in the hippocampus (n=10, 10). Figure 41B shows the percentage of Iba-1 immunoreactive area in the hippocampus (n=10, 10). Figure 41C shows the percentage of GFAP immunoreactive area in the hippocampus (n=10, 10). All data shown are means ± sem. (by unpaired t-test) * p<0.05, ** p<0.01). These results show a significant decrease in the microglial markers CD68 and Iba-1 in the hippocampus of aged mice treated with PPF1.

[0228] (s) Example 19 Commercially available PPF ("PPF1") or saline control was administered to 23-month-old wild type (C57BL / 6J, "WT", strain code 0664, Jackson Labs, Bar Harbor, ME). All mice received 50 mg / kg BrdU for the first week prior to pulse dosing for 7 consecutive days. All mice then received IV injections of 150 μL of PPF1 or saline per dose for 7 consecutive days. One cohort from each treatment group was used to examine histological markers of neuroinflammation and were sacrificed 6 weeks after the last dose administered.

[0229] Figure 42A shows the percent change in BrdU expression, an indicator of cell survival, in the hippocampus compared to saline controls at 6, 9, and 12 weeks after dosing. Mice were treated with a pulsed dosing regimen for 7 consecutive days.

[0230] Figure 42B shows the percent change in doublecortin (DCX), an indicator of neurogenesis in the hippocampus, compared to saline controls at 6, 9, and 12 weeks after dosing. Mice were treated with a pulsed dosing regimen for 7 consecutive days.

[0231] (t) Example 20 Thirty male alpha-synuclein transgenic mice (line 61, wild type background C57BL / 6J), aged 4-4.5 months, were divided into two groups of 15 mice and treated with either PPF1 or vehicle for 7 consecutive days. PPF1 treatment was administered IV at 5 μL per gram of body weight. Alpha-synuclein mice serve as a transgenic model of Parkinson's disease and overexpress the alpha-synuclein protein. This transgenic model, unlike NSG mice, is not immunodeficient.

[0232] The day after the last treatment with PPF1 or vehicle, all mice were subjected to behavioral and motor function tests, such as nest building, pasta chewing, wire suspension, rotarod, and beam walking. Pasta chewing, wire suspension, and beam walking were performed twice at the end of the test. Tests were performed in a randomized order.

[0233] Mice were weighed weekly. Figures 43A and 43B show that there was no significant difference between PPF1-treated and vehicle-treated alpha-synuclein transgenic mice ("Tg").

[0234] Figure 44 shows the results for nest building. Mice were individually housed in cages containing wood chip bedding and one pressed cotton square (nesting material). No other nesting material (e.g., wood wool) was present. The day before nest condition was assessed, nesting material was introduced approximately 2-3 hours before the onset of the dark period, and nesting behavior was assessed approximately 2-3 hours after the onset of the light period the day after the experiment. The length of time between the introduction of the cotton square and the assessment of nest condition was the same for all experiments. Manipulation of nest material and construction of the nest were assessed according to a 5-point scale (Deacon, RM 2006, Assessing nest building in mice. Nat. Protoc. 1:1117-19.). As shown in Figure 44, there was a trend toward increased nest building behavior in PPF1-treated mice compared to vehicle-treated mice.

[0235] Figures 45A and 45B show that 3 weeks after the last treatment, there was a significant increase in pasta chewing in the PPF1-treated group compared to the vehicle-treated group, indicating an improvement in locomotion (Figure 45B). The test was developed to study motor disorders in small rodents. Mice were placed in the experimental room at least 2 hours before the test. The cage lid, water bottle and food pellets were removed and a small piece of dry spaghetti (approximately 5 mm) was placed in the cage. A microphone was placed on the spaghetti piece. Recording began as soon as the mouse started to eat. The number of bites per chewing episode and the frequency of bites were assessed and chewing patterns were analyzed using Avisoft SASLab Pro software. All data shown are mean ± sem. (An unpaired t-test was used to determine the mean ± sem.) * p<0.05).

[0236] FIG. 46 shows the results of the wire mesh suspension test to evaluate neuromuscular abnormalities of motor strength. Three weeks after the last treatment, there was a significant increase in the latency to fall in the PPF1-treated group compared to the vehicle-treated group. To perform the test, a wire mesh cage lid was used with duct tape placed around the perimeter to prevent the mouse from leaving the ledge. The mouse was placed on the cage lid. The lid was gently shaken three times to force the mouse to grasp the wire mesh, and then the lid was turned upside down. The lid was held at a height of approximately 50-60 cm on a soft underlay high enough to prevent the mouse from jumping off, but not so high that it would fall and cause injury. The latency to fall was quantified and a 300 second cutoff time was used. Wild-type mice can normally hang upside down for several minutes.

[0237] Figures 47A, 47B and 47C show the results of the beam walking test. Figure 47A shows the different beam shapes and sizes (square or cylindrical beams) used in five different trials of increasing difficulty. Figure 47B shows the results of five trials 72 hours after the last treatment. Figure 47C shows the results of five trials 3 weeks after the last treatment. Mice treated with PPF1 showed significantly higher success in traversing the beam in trial 5 of test 1 (72 hours after treatment) and trial 4 of test 2 (3 weeks after treatment). All data shown are means ± sem. (by binomial test) ** p<0.01).

[0238] Figures 48A-48F show histological results of staining of the striatum and hippocampus. Figure 48A shows staining of CD68 in the striatum. Figure 48B shows staining of CD68 in the hippocampus. Figure 48C shows staining of Iba-1 in the striatum. Figure 48D shows staining of Iba-1 in the hippocampus.

[0239] FIG. 48E shows NeuN staining in the striatum. FIG. 48F shows NeuN staining in the hippocampus. These figures show that mice treated with PPF1 demonstrated reduced microgliosis (neuroinflammation) in both the striatum and hippocampus by Iba-1 and CD68, and increased neuronal survival by NeuN in the striatum and hippocampus. All data shown are means ± sem. (Paired t-test)* p<0.05, ** p<0.01, *** p<0.001).

[0240] (u) Example 21 PPF1, HAS1 or saline control was administered to 12-month-old mice (NOD.Cg-Prkdscid Il2rgtm 1 Wj1 / SzJ, "NSG" strain). HAS1 is a commercially available HAS prepared by cold alcohol fractionation and derived from pooled human plasma from donors, which is >95% human albumin (relative to total protein) in a 5% solution (w / v, 50 g / L). Except where noted, HAS1 was administered in vivo using 5% in the examples herein. Mice were injected IV with PPF1, HAS1 or sterile saline at 150 μL per dose daily for 7 consecutive days and analyzed behaviorally 4 weeks after the last dose.

[0241] FIG. 49 shows the time it took for mice to enter the escape hole for PPF1, HAS1 and vehicle treated mice to escape the Barnes maze. PPF1 treated mice found the escape hole significantly faster than vehicle treated mice. The data show that PPF1 effectively improves cognition in aged NSG animals while HAS1 has no effect on hippocampus-dependent memory. All data shown are mean±sem. (Paired t-test) * p<0.05).

[0242] (v) Example 22 Clinical norms using the PPF (1) Mild to moderate AD Men and women aged 60 years or older with mild to moderate AD will be randomly assigned to receive 100 mL or 250 mL PPF1 ("pulse dosing") once daily for 5 days during weeks 1 and 13 of the study for a total duration of 6 months. Between the two 5-day dosing periods, subjects will reside in an inpatient observation unit to facilitate safety assessments, and all subjects will have a screening visit, a baseline visit, a treatment visit, a follow-up visit, and an end-of-study visit / early termination visit. Safety and tolerability assessments will occur at every visit. Neurocognitive and motor assessments will be performed at baseline and at regular interim visits following dosing.

[0243] The primary endpoints are the safety, tolerability, and feasibility of each dosing regimen. Safety will be measured by the occurrence of treatment-emergent adverse events. Tolerability will be measured by the number of subjects completing after 8 weeks having received at least 5 infusions and the number of subjects completing after 24 weeks having received at least 10 infusions. Study feasibility will be measured by the number of subjects completing 5 and 10 infusions. Secondary endpoints will assess possible effects on cognition using a variety of established cognitive assessment measures, including the Alzheimer's Disease Assessment Scale-Cognitive Subscale. Exploratory endpoints include evaluation of changes in magnetic resonance imaging as well as changes in the composition and distribution of blood-based biomarkers.

[0244] (2) Mild to moderate AD Two groups of subjects diagnosed with mild to moderate AD will be randomized to active treatment in a double-blind fashion. All subjects will receive a single infusion per day on 5 consecutive days of randomized dosing in weeks 1 and 13 for a total study period of 6 months. Subjects will be randomized to one of two dose levels of PPF1, 100 mL and 250 mL. Dosing groups will also be stratified by gender. Administration time will be 2-2.5 hours, with flow rates set according to administration-specific guidelines to ensure the entire dose is administered.

[0245] Subjects will participate in an optimal CSF biomarker study. Such subjects will undergo two lumbar punctures to obtain CSF, the first prior to the first dose and the second following the final dose. Neurocognitive and motor assessments will be performed at baseline and at regular interim assessments following dosing.

[0246] The safety, tolerability, and feasibility of each dosing regimen will be determined. Cognitive scores will be determined and summarized for tests including the Mini-Mental State Examination (MMSE), Alzheimer's Disease Assessment Scale-Cognitive Subscale 11-item (ADAS-Cog / 11), Grooved Pegboard Test, Category Fluency Test (CFT), Clinical Dementia Rating Scale-Sum of Boxes (CDR-SOB), Alzheimer's Disease Cooperative Study-Activities of Daily Living (ADCS-ADL), Alzheimer's Disease Cooperative Study-Clinical Global Impression of Change (ADCS-CGIC), Neuropsychiatric Inventory Questionnaire (NPI-Q), and Savonix Neurocognitive Assessment and Digit Span.

[0247] (3) Parkinson's disease Subjects with Parkinson's disease and cognitive impairment are randomized into two groups, active treatment and placebo. Subjects receive a daily infusion of active treatment or placebo treatment for 5 consecutive days ("pulse dosing") during the first week of the study. During week 13, both groups receive active treatment for 5 consecutive days, and the study duration is approximately 7 months. Administration time is 2-2.5 hours, and flow rate is set according to administration-specific guidelines to ensure the entire dose is administered.

[0248] The safety, tolerability and feasibility of each dosing regimen will be determined. Cognitive and motor function will be summarized across tests including MoCA, attention, working, memory and episodic memory persistence and performance in CDR-CCB, MDS-UPDRS3, MDS-UPDRS2, SE-ADL, and CISI-PD.

[0249] It is to be understood that the invention is not limited to the particular embodiments described, as such may vary. The scope of the present invention will be limited only by the appended claims, and it is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

[0250] Where a range of values ​​is provided, each intervening value between the upper and lower end of that range, to the tenth of the unit of the lower end unless the context clearly dictates otherwise, and any other stated or intervening value in that stated range, is encompassed within the invention. Subject to any specifically excluded end in a stated range, the upper and lower ends of these smaller ranges may independently be included in the smaller ranges and are also encompassed within the invention. Where the stated range includes one or both of the ends, ranges excluding either or both of those included ends are also included in the invention.

[0251] 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 also be used in the practice and testing of the present invention, representative illustrative methods and materials are described herein.

[0252] The publications and patents cited herein are incorporated by reference as if each publication or patent was specifically and individually indicated to be incorporated by reference herein and as if incorporated by reference to disclose and describe the methods and / or materials for which the publications are cited. The citation of a publication is for its disclosure prior to the filing date and should not be construed as an admission that the present invention is not entitled to antedate such publication by reason of prior invention. Further, the dates of publication provided may be different from the actual publication dates which may need to be independently confirmed.

[0253] As used in this specification and the appended claims, it is noted that the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. It is further noted that the claims may be drafted to exclude any element. As such, this reference is intended to serve as a predicate for the use of exclusionary terminology such as "solely," "only," and the like in conjunction with recitation of claim elements or the use of a "negative limitation."

[0254] As will be apparent to one of skill in the art upon reading this disclosure, each of the individual embodiments depicted and illustrated herein has separate components and features which may be readily separated from or combined with the features of any of the other several embodiments without departing from the scope or spirit of the invention. Recited methods can be carried out in the order of events recited or in any other order which is logically possible.

[0255] Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be readily apparent to those skilled in the art that, in view of the teachings of this invention, certain changes and modifications may be made thereto without departing from the spirit or scope of the appended claims.

[0256] Thus, the foregoing merely illustrates the principles of the invention. It is well understood that those skilled in the art can devise various arrangements which, although not expressly described or shown herein, embody the principles of the invention and are included within its spirit and scope. Furthermore, all examples and conditional language recited herein are intended primarily to aid the reader in understanding the invention and the concepts contributed by the inventor to the broadening of the art, and should not be construed as being limited to such specifically recited examples and conditions. Furthermore, all references herein reciting principles, aspects, and embodiments of the invention as well as specific examples thereof are intended to encompass their structural and functional equivalents.

[0257] Additionally, it is intended that such equivalents include both currently known equivalents as well as equivalents developed in the future, i.e., elements developed that perform the same function, regardless of structure. Thus, the scope of the present 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.

[0258] Related Applications Pursuant to 35 U.S.C. 35(e), this application claims priority to the filing dates of U.S. Provisional Patent Application No. 62 / 490,519, filed April 26, 2017, U.S. Provisional Patent Application No. 62 / 584,571, filed November 10, 2017, U.S. Provisional Patent Application No. 62 / 623,468, filed January 29, 2018, and U.S. Provisional Patent Application No. 62 / 641,194, filed March 9, 2018, the disclosures of which are incorporated herein by reference.

Claims

1. A blood plasma preparation for treating a subject diagnosed with cognitive impairment with a pulsed dosing regimen, comprising: an effective amount of a plasma protein fraction (PPF) which is a sterile solution of proteins composed of albumin and globulin derived from human plasma; The PPF contains 83% or more but less than 95% albumin based on the total protein contained in the PPF, 17% or less globulin and other plasma proteins based on the total protein contained in the PPF, and 1% or less gamma globulin based on the total protein contained in the PPF; Blood plasma preparations.

2. The blood plasma preparation of claim 1, wherein the PPF is a commercially available PPF.

3. The blood plasma preparation of claim 1 or 2, wherein the PPF is derived from plasma from a pool of young individuals.

4. The blood plasma preparation according to any one of claims 1 to 3, wherein the PPF is produced from a mammalian blood product.

5. The blood plasma product of claim 4, wherein the mammalian blood product is a human blood product.

6. The blood plasma preparation according to any one of claims 1 to 5, wherein the subject is a mammal.

7. The blood plasma preparation of claim 6, wherein the mammal is a human.

8. The blood plasma preparation of any one of claims 1 to 7, wherein the pulsed dosing regimen comprises administration of the PPF for 5 to 7 consecutive days.

9. The blood plasma preparation of any one of claims 1 to 8, wherein the subject is subjected to an exercise stress regimen after complete administration of the pulsed dosing regimen.

10. 1. A kit for use in treating a subject for cognitive impairment with a pulsed dosing regimen, comprising: A kit comprising the blood plasma preparation according to any one of claims 1 to 5.

11. The kit of claim 10, wherein the pulsed dosing regimen comprises administration of the PPF for 5 to 7 consecutive days.

12. 12. The kit of claim 10 or 11, wherein the subject is subjected to an exercise stress regimen after complete administration of the pulsed dosing regimen.

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