Factor VIII-RAG From Human Endothelial Cells
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Abstract:
Factor VIII related antigen (VIII-RAG) can be demonstrated in cultured human endothelial cells (EC) from umbilical cord by immunofluorescence. 75Se-Methionine tracer added onto culture medium showed that radioactive 75Se was incorporated into VIII-RAG as evidenced by Laurell Crossed Immunoelectrophoresis (CIE) and autoradiograph. This indicates that EC synthesizes VIII-RAG in culture, confirming the previous findings of Jaffe et al (1973) and Shearn et al (1977). 75Se- VIII in medium had similar CIE pattern as normal plasma VIII, normal ristocetin cofactor activity (RCoF), but no procoagulant (VIII-C) activity. However, 75Se-VIII in cell sonicate (EC-VIII) migrated faster on electrophoresis and had no RCoF or VIII-C activities. The EC-VIII was compared to factor VIII dissociated into subunits by cleavage of SH bonds. It can be concluded that VIII-RAG is synthesized as monomers in EC and polymers are formed on secretion. Hence, defect in VIII-RAG may arise from defective synthesis or defective assembly mechanism in the EC.Keywords:
Cleavage (geology)
Immunoelectrophoresis
Neonatal tetanus infection and umbilical cord infection (omphalitis) are the main causes of morbidity and mortality in infants throughout the country, especially in Southeast Asia due to improper umbilical cord care. Umbilical cord care is carried out after the neonate is born until the umbilical cord is released with great care, attention and care to prevent infection in the umbilical cord. The purpose of this study was to describe the results of implementing the implementation of umbilical cord care with the open method in neonates. This type of research method uses a case study by applying the open method of umbilical cord care to determine the timing of the release of the umbilical cord. The results of the condition of the neonate's umbilical cord Ny. A and Mrs. T has not been released, before the umbilical cord is treated with an open method. While the condition of the neonate's umbilical cord, Ny. A and Mrs. T has been released, after the open method of umbilical cord treatment with the release of the umbilical cord in the neonate Ny. A was released on the 4th day, quickly (umbilical cord detachment time <5 days) and the neonate Mrs. T loose on day 5, normal (time of umbilical cord detachment 5-7 days). Based on the results of the final evaluation, there are differences in the development of the umbilical cord release time before and after being given the open method of umbilical cord care.
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To investigate the influencing factors for the time of umbilical cord separation in full-term newborns.The time of umbilical cord separation was recorded in 337 full-term newborns. Single factor and multifactor unconditioned logistic regression were performed to investigate the influencing factors of umbilical cord separation. Fourteen possible factors associated with the time of umbilical cord separation, including sex, gestational age, body weight, position of umbilical cord ligature, length of umbilical cord stump, umbilical cord diameter, cleanness of umbilical cord paster, hand cleanness of medical staff and family members and umbilical infection, were involved.The single factor correlative analysis demonstrated that the position of umbilical cord ligature, length of umbilical cord stump, umbilical cord diameter, cleanness of umbilical cord paster, and umbilical infection were influencing factors for the time of umbilical cord separation (P<0.05). The multifactor unconditioned logistic regression analysis demonstrated four major influencing factors for umbilical cord separation: position of umbilical cord ligature, length of umbilical cord stump, cleanness of umbilical cord paster, and umbilical infection.The following factors contribute to early separation of umbilical cord: the proper position of umbilical cord ligature (<0.5 cm to umbilical ring), the umbilical cord stump of <0.5 cm, keeping the umbilical cord paster clean and the prevention of umbilical infection.
Ligature
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Immunological and physicochemical properties were compared for water-soluble acid neurospecific antigens: D of the bull and alpha of the rat. During immunoelectrophoresis in agar gel antigen D migrates as two immunologically indentical fractions having a position of prealbumins and alpha-globulins of blood serum. Antigen D from the bull brain is incompletely immunologically identical with antigen alpha from the rat brain. The molecular weight of antigen D determined under conditions of Sephadex G-100 column gel chromatography is 73000.
Sephadex
Immunoelectrophoresis
Immunodiffusion
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Summary About 12 antigens were demonstrated in saline extracts of homogenized rat brain by immunodiffusion and immunoelectrophoresis. A brain organ antigen was discovered which appears to be species restricted. One antigen was found to be shared by similar extracts of bovine, guinea pig, human or rabbit brains. Immunoelectrophoretic analyses of the brain extracts showed this organ antigen had an α2 or β1 mobility, depending on the species from which it was derived. About nine antigens were species-specific globulins and could be partially separated from one another by diethylaminoethyl cellulose chromatography. All of these antigens were shared by the brain and different rat organs. However, one major antigen was shared by all the rat organs tested.
Immunoelectrophoresis
Immunodiffusion
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A successful isolation of Mesenchymal Stem Cells (MSCs) for its clinical application is very important.Isolation of MSCs from neonatal sources like human Umbilical Cord Wharton's Jelly (hUCWJ) have been reported in many studies and are considered primitive but isolation of MSCs from human umbilical cord blood (hUCB) is open to discussion. MATERIALS AND METHODSThe MSCs from hUCWJ and hUCB were isolated by explant culture and Ficoll density gradient respectively.The media used for the two sources were Dulbecco's modified Eagles medium, GlutaMax and fetal bovine serum (FBS) for hUCWJ, and Iscove modified Dulbecco's medium and FBS for hUCB.Further, the isolated cells were studied as per criteria of International Society of Cellular Therapy to define the cells as MSCs.Finally, we compare the feasibility and ease of isolating the neonatal sources. RESULTSUnlike hUCWJ, defining hUCB cells to MSCs as per criteria is limited because the number of cells decreases once it reaches 50% confluence and after 2 weeks, they show differing morphology with an appearance of large dimension of multinucleated cells identifying them as osteoclast-like cells thus restricting further study.The cells derived from both the sources grew into long, spindle shaped cells with prominent nuclei but hUCB has limited proliferation and fulfilled only the first criterion to define as MSCs while the cells isolated from hUCWJ continue to grow for multiple passages and fulfil all the defining criteria. CONCLUSIONNeonatal derived hUCWJ is more ideal source for isolation of MSCs than hUCB with the potential to use it in regenerative medicine.
Wharton's jelly
Cord lining
Placenta cord banking
Cord blood
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It is generally accepted that there is a need for standardized antigens in the diagnostic of exogenous allergic alveolitis. For this reason, ten antigens in a pigeon intestinal extract were classified in major, minor and intermediate antigens according to the distribution of specific antibodies in sera of asymptomatic and ill persons by use of crossed immunoelectrophoresis. Using a reference antiserum with antibodies against all ten detected antigens it is possible to prove a reproducible antigen composition in following preparations.
Immunoelectrophoresis
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Immunoelectrophoresis
Antibody response
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Summary 1 )About 12 antigens were demonstrated in saline extracts of homogenized rat brain by immunodiffusion and immunoelectrophoresis. 2 )A brain organ antigen was discovered which appears to be species restricted. 3 )One antigen was found to be shared by similar extracts of bovine, guinea pig, human or rabbit brains. Immunoelectrophoretic analyses of the brain extracts showed this organ antigen had an α 2 or β 1 mobility, depending on the species from which it was derived. 4 )About nine antigens were species-specific globulins and could be partially separated from one another by diethylaminoethyl cellulose chromatography. All of these antigens were shared by the brain and different rat organs. However, one major antigen was shared by all the rat organs tested.
Immunoelectrophoresis
Immunodiffusion
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By means of the antigen-antibody crossed electrophoresis procedure of Laurell, 68 antigens were demonstrated in Candida albicans . This is about four times the number of antigens described earlier by means of classical immunoelectrophoresis. The procedures for obtaining this result are described, including the preparation of antigen, the immunization of rabbits, and the method of N. M. G. Harboe for the production of purified and concentrated rabbit antibodies suitable for quantitative immunoelectrophoresis. The immunoplates were stained by means of the sensitive Coomassie brilliant blue R. The various quantitative immunoelectrophoretic methods offer considerable possibilities for qualitative and quantitative characterization of antigens, even in complex mixtures, and are therefore well suited for the investigation of microbial antigens.
Coomassie Brilliant Blue
Immunoelectrophoresis
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The development of the human umbilical cord blood transplants with hematopoietic repopulating cells has enabled some problems associated with bone marrow transplants to be solved. Frozen umbilical cord blood banks should facilitate the finding of suitable stem cell donors. However, further experience is necessary to develop the optimal method for collection, separation, storage and cryopreservation of umbilical cord blood. We report our experience in the organization of a Cord Blood Bank.
Cord blood
Placenta cord banking
Hematopoietic stem cell
Blood units
Cord lining
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