Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P30536 (PBS)
 9,886 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

One hundred sixty-two embryos were collected from superovulated crossbred beef cattle 7 to 8 d after the onset of estrus. Embryos were frozen in modified Dulbecco's phosphate-buffered saline supplemented with 20% heat-inactivated fetal calf serum (PBS + FCS) and dimethylsulfoxide (DMSO), which was added in three steps to a final concentration of 1.5 M. Embryos were placed in .25 ml of 1.5 M DMSO in PBS + FCS in 1-ml glass ampules and cooled at 1.0 C/min from ambient temperature to -7 C, seeded and then cooled at .3 C/min to -19, -26, -33, -38, -43, -50 or -57 C before immersion (plunging) in liquid nitrogen. Ampules were thawed in 25 C water, and DMSO was removed in six steps at .25 M increments. 10 min/step. After removal of DMSO, embryos were cultured 24 h in PBS + FCS and then fixed and stained. Just after thawing, embryos for which slow cooling was terminated at -50 C were of lower (P less than .05) morphological quality than other groups. After removal of cryoprotectant, embryos from both the -19 and -50 C treatments had deteriorated more (P less than .05) than had embryos from other treatments. After 24-h culture, embryos slow-cooled to -19, -26 and -50 C had a lower rate of survival (P less than .05) than did embryos from -33, -38, -43 and -57 C temperatures. Embryos slow-cooled to -33, -38 and -43 C showed a higher percentage of healthy nuclei than did embryos slow-cooled to -19, -26 and -50 C.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Effect of slow cooling end point temperature on survival of frozen bovine embryos. 404 43

We describe the development of a simple and highly sensitive double antibody sandwich enzyme-linked immunosorbent assay (ELISA) for measuring IgG and IgM anticardiolipin antibodies (ACA). Microtitre plates were coated with cardiolipin at a concentration of 45 micrograms/ml by evaporation under nitrogen. Non-specific binding of diluted sera was eliminated by blocking of plates with 10% fetal calf serum in phosphate buffered saline (PBS/FCS) for 2 h. Then sera (100 microliters) at a dilution of 1:100 were incubated in the wells for 1 h. Affinity purified goat anti-human IgG or IgM (100 microliters) at a concentration of 1 microgram/ml was subsequently added and allowed to incubate for 1 h; detection of ACA was achieved using an alkaline phosphatase conjugated rabbit anti-goat IgG reagent by reading the colorimetric yield at 405 nm after incubation with substrate. Reference serum pools were established to study reproducibility of the assay throughout its sensitivity range, and Standard curves were established. The quantitative normal range was 0-9.0 Anticardiolipin ELISA Units (AEU) for IgG and 0-8.0 (AEU) for IgM-ACA. A strong correlation was found between the ELISA and radioimmunoassay methods for measuring ACA of both IgG and IgM classes. Results from 65 patients with systemic lupus erythematosus (SLE) and 45 patients with seropositive rheumatoid arthritis are also reported. The advantages of the ELISA method for quantitative determination of ACA levels, should make it a useful and reliable method for clinical and experimental monitoring of patients with SLE and associated autoimmune disorders.
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PMID:Measurement of anti-cardiolipin antibodies by an enzyme-linked immunosorbent assay (ELISA): standardization and quantitation of results. 408 54

Mutants have been isolated which correspond to every step concerned with the biosynthesis of the aromatic amino acids in Bacillus subtilis. Each mutant has been characterized, and the lesion it bore was analyzed by deoxyribonucleic acid transformation and PBS-1 mediated transduction. The biochemical analysis revealed that each of the mutations appears to have affected a single enzyme, except for two groups of pleiotropic mutations. All aroF mutants (chorismic acid synthetase) lack dehydroquinic acid synthetase (aroB) activity. The gene that specifies aroB is closely linked to the gene coding for the aroF enzyme. Both genes are a part of the aro cluster. Mutants lacking chorismate mutase activity also lack d-arabino-heptulosonic acid-7-phosphate synthetase and shikimate kinase activity, presumably as a result of these three activities forming a multi-enzyme complex. Another mutant, previously undescribed, had been isolated. The affected gene codes for the tyrosine and phenylalanine aminotransferase activity. All of the mutations have been located on the B. subtilis genome except those in the genes specifying shikimate kinase activity and tyrosine-phenylalanine aminotransferase activity.
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PMID:Gene-enzyme relationships of aromatic acid biosynthesis in Bacillus subtilis. 420 Aug 44

The use of three diluents (i.e., 0.01 m phosphate-buffered saline, PBS; PBS with 0.2% gelatin, PBS/GEL; and PBS with 0.4% bovine plasma albumin) and three methods (i.e., the standard tube macro-procedure, TUBE; the manual microtechnique, MANUAL; and the semiautomatic microtechnique, AUTO) were statistically compared for their reproducibility and sensitivity in determining hemagglutinin (HA) and hemagglutination-inhibition (HI) antibody titers. In the HA test, analyses of between-cell variances of the different methods showed the AUTO microtiter procedure to be more reproducible than the standard TUBE method. The MANUAL microtiter procedure was the least reproducible. In the HI test, the TUBE method was the most reproducible. No significant difference in the reproducibility of the diluents was observed in either the HA or HI test. When a comparison of the sensitivity of test methods and diluents was made for determining HA titers, the AUTO microtiter procedure and PBS/GEL diluent appeared to be the method and diluent of choice. Evaluation of another instrument, the autopipetter, which standardizes the volume of diluent to be added in the microtechnique, suggests that the reproducibility of the AUTO microtiter procedure might be further increased.
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PMID:Statistical evaluation of diluents and automatic diluting and pipetting machines in influenza serology. 554 4

An inhibition enzyme-linked immunosorbent assay (IELISA) was used to detect the presence of schistosome antigens obtained from cercariae, adult worms, and eggs of the parasite. Using appropriate titers of Schistosoma mansoni infected mouse serum (IMS), it was possible to detect less than 10 ng/ml of schistosome antigen when added to phosphate-buffered saline (PBS, pH 7.2) or normal human serum (NHS). The sensitivity of the test was highly contingent on the number of experimental variables including antibody titer and antigenic source. The results of specificity studies were complicated. Although there was no cross-reactivity detected with other unrelated antigen preparations, extensive cross-reactivity between various schistosome species and "stage-specific" antigens was observed. The IELISA, utilizing IMS, can quantitate the degree of antigenic cross-reactivity, i.e., genus-specific and cross-reacting antigenic determinants. Soluble egg antigen (SEA) preparations obtained from S. mansoni and S. japonicum actually "cross-reacted" more than cercarial- and egg-derived antigens obtained from the same species (S. mansoni). This test also showed a 32-fold increase in specificity for the quantitative detection of specific antigenic determinants when monoclonal antibodies were used to restrict the heterogeneity of the measured response. The technique proved satisfactory for the quantification of parasitic burden in mice and the detection of active infections in humans. Circulating antigen disappeared with a t 1/2 of 72-96 hr after successful treatment.
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PMID:Schistosoma mansoni: detection and characterization of antigens and antigenemia by inhibition enzyme-linked immunosorbent assay (IELISA). 618 95

Anterior pituitaries from primiparous lactating C3H/He mice cultured in the medium containing 0.1% dimethyl sulfoxide (DMSO) for 48 hours and pituitaries from lactating mice given subcutaneous injections of 0.05 ml DMSO twice daily for two days and once on the morning of the third day were used in the studies of the in vitro and the in vivo effects of DMSO, respectively. Phosphate buffer saline was used in the control. Synthesis and release of growth hormone and prolactin were estimated by the incorporation of [14C]leucine into each hormone during three hours' incubation of the pituitaries pre-exposed to DMSO or PBS. The values in the medium represented released hormone and sum of the values in the medium and the pituitary represented the synthesized hormone. DMSO stimulated synthesis of GH and synthesis and release of prolactin in vitro. Meanwhile, in the vivo study, synthesis of GH and prolactin were lower in the DMSO-injected mice than in the control. The results suggest that the effects of DMSO on the pituitary secretion of GH and prolactin are adverse in vitro and in vivo. In vivo exposure of pituitary to DMSO resulted in the suppression of lactation.
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PMID:The in vitro and in vivo effects of dimethyl sulfoxide on the pituitary secretion of growth hormone and prolactin in mice. 619 45

We examined the localization of five pregnancy-associated proteins such as pregnancy specific beta 1-glycoprotein (SP1), human chorionic gonadotropin (HCG), human placental lactogen (HPL), alpha 2-PA-glycoprotein (SP3) and pregnancy-associated plasma protein-A (PAPP-A) in the placentae of early and term pregnancies by means of the PAP method in immunohistochemical technology. It was found that the degree of staining of these proteins did not reflect their concentration in serum at each gestational age. Therefore, the concentrations of these proteins in the solution extracted from the placenta were measured. 1) Placental tissues were fixed with a 10% neutral formaldehyde solution and these were embedded into paraffin blocks. These specimens were used for the PAP method. The localizations of five pregnancy-associated proteins from 10 placentae at the gestational age of 7 to 10 weeks were compared with those at the gestation age of 38 to 41 weeks from 7 patients. The staining degree of SP1 in free villi was pale in both early and term pregnancies. HCG was stained deeply in early pregnancy but pale at term pregnancy. HPL was stained deeply at both gestational ages. Horseradish peroxidase reaction products from SP1, HCG and HPL were located chiefly in the syncytiotrophoblast. SP3 was not stained on the placental tissue. PAPP-A was stained to a greater extent in the cytotrophoblast in early pregnancy but pale in the syncytiotrophoblast at term pregnancy. 2) The concentrations of SP1, HCG, HPL, SP3 and PAPP-A were measured in the placental tissues in both early and term pregnancies. Placental tissues were obtained from 24 normal pregnancy patients aborted artificially at 8 to 11 weeks gestation and from 28 patients terminated by normal deliveries at 38 to 40 weeks gestation. The tissue was homogenated with 3 volumes of 1/2 PBS (0.005M phosphate-buffered 0.07M sodium chloride, pH 7.4). The supernatant was removed after centrifugation and stored at -20 degrees C until assayed. The concentration of SP1 was measured by single radial immunodiffusion. That of HCG was assayed by a directed Latex agglutination test (Gestate Slide Eiken). The concentrations of HPL, SP3 and PAPP-A were quantified by rocket immunoelectrophoresis. No significant difference in SP1 levels was shown between early and term pregnancies, and the SP1 level was 4.3 +/- 1.3 (mean +/- 1 S.D.) mg/dl at term pregnancy. The HCG level was 1,100 +/- 300 IU/ml in early pregnancy and at least 20-fold higher than that at term pregnancy.(ABSTRACT TRUNCATED AT 400 WORDS)
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PMID:[Placental tissue concentration of pregnancy-associated proteins in early and term pregnancies]. 619 24

We studied two patients with an axonal type of polyneuropathy, epidermolysis, and IgM kappa plasma cell dyscrasia. The IgM kappa was deposited in the dermis, was absorbed from the serum by axonal micelle preparations, and was precipitated with chondroitin sulfate in highly purified agarose in 0.15 M NaCl with 0.01 M phosphate buffer, pH 7.8. In contrast, we found none of these abnormalities in three patients with IgM plasma cell dyscrasia and demyelinating neuropathy. Of 78 other macroglobulinemic serum samples from patients without neuropathy, 7 precipitated with a sulfated polysaccharide. This reaction occurred at low ionic strength, 0.05 M barbital buffer, pH 8.1, but did not occur in the higher ionic strength of 0.01 M phosphate with 0.15 M NaCl (PBS). The interaction of the IgM with chondroitin sulfate at relatively high ionic strength could cause both the axonal polyneuropathy and the epidermolysis.
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PMID:Monoclonal IgM kappa antibody precipitating with chondroitin sulfate C from patients with axonal polyneuropathy and epidermolysis. 629 26

The possibility that prostaglandin E2 (PGE2) increases endometrial vascular permeability and initiates decidualization in sensitized rat uteri by stimulation of adenosine 3':5'-cyclic monophosphate (cAMP) synthesis was investigated. Immature rats, pretreated so that they were sensitized for the decidual cell reaction, were used. Following the unilateral intrauterine injection of 50 microliters phosphate-buffered saline containing gelatin (PBS-G), a deciduogenic stimulus, uterine concentrations of both PGE and cAMP were elevated as early as 1 min after the intrauterine treatment. To determine if uterine stimuli which increase endometrial vascular permeability also increase uterine cAMP concentrations, rats, treated with or without indomethacin, an inhibitor of PG synthesis, received unilateral intrauterine injections of 50 microliters PBS-G with and without 10 micrograms PGE2 and were killed 15 min later. Uterine cAMP concentrations were elevated in all injected horns except in those of indomethacin-treated rats receiving PBS-G intraluminally, thus paralleling the expected changes in endometrial vascular permeability. As indicated by radioactivity levels in the stimulated horn 15 min after the i.v. injection of 125I-labeled bovine serum albumin, the intrauterine injection of dibutyryl cAMP, with or without theophylline, did not increase endometrial vascular permeability in indomethacin-treated animals. In contrast, cholera toxin, an activator of adenylate cyclase activity, markedly elevated permeability and induced decidualization. Except for the lack of a permeability response to the cAMP analogue, these data are consistent with the hypothesis that the effect of PGE2 on endometrial vascular permeability is mediated by cAMP.
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PMID:Prostaglandin E2, adenosine 3':5'-cyclic monophosphate and changes in endometrial vascular permeability in rat uteri sensitized for the decidual cell reaction. 631 67

The aggregates of washed control and NANAse-treated human red blood cells (RBC) were characterized by scanning electron microscopy (SEM). Aggregation of RBC was performed under well-defined laminar shear conditions (78.6 S-1) in a horizontally arranged coaxial cylinder system. Generally, with increasing La3+ concentration in the PBS the aggregates became larger and the contact between the RBC "more intensive". This was evidenced by more cells per aggregate, an enlargement of contact areas, a change of cell shape in the contact region and signs of fusion phenomena. Especially at high La3+ concentrations on the cell surface of treated and untreated RBC some deposits were seen by SEM which are possibly La3+- phosphate precipitates. Neuraminidase treatment promoted the described La3+ effects. Concanavalin A-induced RBC agglutinates were different from the more spherical former ones and showed a network-like morphology. The conclusions drawn on the basis of light and scanning electron microscopical investigations are confirmed by quantitative values of an aggregation index estimated in parallel.
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PMID:Scanning electron microscopical characterization of La3+- and concanavalin A-induced aggregations of untreated and neuraminidase-treated human erythrocytes. 733 76


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