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Glass laminate structures for head-up display system

A technology of protein expression and detection method, which is applied in the field of detection of protein expression in live motor neurons, and can solve problems such as obstacles to the establishment of fundamental treatments for SMA

Active Publication Date: 2016-12-21
TOKYO WOMENS MEDICAL UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] As described above, in the prior art, there are various problems in order to use SMN protein as a useful biomarker for SMA with onset infancy, and the establishment of a fundamental treatment for SMA has become a major obstacle.
[0013] In addition, using blood samples such as whole blood (peripheral blood) or a substance obtained by hemolyzing erythrocytes in whole blood, there has been no report so far that a difference in the expression level of SMN protein between a normal person and an infantile-onset SMA patient can be detected. A reliable and reliable detection method for SMN protein

Method used

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  • Glass laminate structures for head-up display system
  • Glass laminate structures for head-up display system
  • Glass laminate structures for head-up display system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0094] Example 1: Detection of SMN Protein Expression Using Lymphoblasts

[0095] (1) Lymphoblastization treatment

[0096] Blood samples obtained from normal persons were sampled in heparin tubes, superimposed on Ficoll solution, centrifuged at 2000 rpm for 15 minutes, and peripheral blood mononuclear cells were separated and sampled. Next, lymphoblasticization was performed by using the transformation method with Epstein-Barr virus.

[0097] Blood samples obtained from type I SMA patients were also treated in the same manner as blood samples obtained from normal individuals.

[0098] (2) Determination of SMN protein expression

[0099] In a centrifuge tube, the normal human-derived peripheral blood mononuclear cell lymphoblasts obtained in the above (1) were sampled, and the cells were fixed with 4% p-formaldehyde-phosphate buffer. Next, the cells were washed with a phosphate buffer solution, centrifuged at 500×g for 5 minutes, and the supernatant was removed. As a cell ...

Embodiment 2

[0116] Embodiment 2: Use the detection of the SMN protein expression that has hemolysis method

[0117] (1) Hemolysis treatment

[0118] Blood was collected from a normal person, and 2 mL of blood was sampled in a heparin tube and mixed by inversion. Next, take 0.5 mL of blood in a centrifuge tube as a hemolysis / fixation solution, and add BD Phosflow TM Lyse / Fix Buffer (BD Biosciences), after standing at 37°C for 10 minutes, centrifuge at 2300rpm for 8 minutes, remove the supernatant, wash the cells with phosphate buffer, and obtain a sample containing nucleated cells derived from blood .

[0119] Carriers and type I SMA patients were also prepared samples containing nucleated cells derived from blood by the hemolysis method in the same manner as normal persons.

[0120] (2) Determination of SMN protein expression

[0121] Add BD Phosflow to normal human nucleated cells obtained from the above (1) TM Perm Buffer II (BD Biosciences) was used as a cell membrane permeation...

Embodiment 3

[0129] Example 3: Detection of SMN protein expression per cluster after classification using surface antigen markers

[0130] Prepare samples according to the staining protocol below.

[0131] 1. Put 2 mL of peripheral blood obtained from a normal person into a tube, and add reagents to 2 mL of peripheral blood to make 100 μL of Hoechst 33342 (Molecular Probes) diluted to 5 μg / mL with PBS and Fc receptor blocking reagent (ClearBack, MTG -001,MBL) 80 μL.

[0132] 2. Plug the vial and rotate slowly at room temperature for 15 minutes in the dark.

[0133] 3. Put 500 μL of the thus treated peripheral blood into each of four 15 mL conical tubes.

[0134] 4. Add 5 μL of fluorochrome-labeled antibody (purchased from BioLegend, BD Biosciences, and MERCK) to each tube according to the following Table 5.

[0135] [table 5]

[0136]

[0137] Cloning of monoclonal antibodies:

[0138] CD66a / c / e(ASL-32),CD66b(G10F5),CD34(581),CD25(M-A251),CD33(WM53),

[0139] CD123(6H6),CD3(UCHT1)...

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Abstract

A glass laminate structure comprising a non- strengthened external glass sheet, a strengthened internal glass sheet, and at least one polymer interlayer intermediate the external and internal glass sheets. The internal glass sheet can have a thickness ranging from about 0.3 mm to about 1.5 mm, the external glass sheet can have a thickness ranging from about 1.5 mm to about 3.0 mm, and the polymer interlayer can have a first edge with a first thickness and a second edge opposite the first edge with a second thickness greater than the first thickness. Other embodiments include external and internal strengthened glass sheets as well as an external strengthened glass sheet and an internal non- strengthened glass sheet.

Description

technical field [0001] The present invention relates to a method for detecting the expression of live motor neuron (SMN) protein. Background technique [0002] Spinal muscular atrophy (SMA) is a muscular atrophy caused by lesions of the anterior horn cells of the spinal cord, and exhibits lower motor neuron symptoms characterized by decreased muscle strength and muscle atrophy of the trunk or limbs. SMA is divided into types I to IV according to the age of onset and severity. Type I with onset at 6 months after birth: severe type (also called Vejo's palsy), type II with onset at 1 year and 6 months: intermediate type (also known as Dubowitz disease), and type III with onset after 1 year and 6 months: mild type (also known as Kugelberg-Welander disease) is SMA with onset in early childhood. On the other hand, type IV that develops after the age of 20 is adult-onset SMA. [0003] Type I SMA accounts for about 30% of SMAs, with onset under 6 months of age. Its symptoms are e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N21/64G01N33/48G01N33/49C07K14/435
CPCG01N33/48G01N21/6428G01N2021/6439G01N33/56972G01N2800/2878G01N33/6875G01N33/6896C07K14/435G01N33/582G01N2333/47
Inventor 斋藤加代子荒川玲子荒川正行野本明男
Owner TOKYO WOMENS MEDICAL UNIV