Role of calcium in the histamine release induced by D-galactosamine from rat mast cells

Rat peritoneal mast cells were isolated and purified by differential centrifugation in Ficoll. Cells pooled from three to four rats were suspended at approximately 10(6) cells/ml in a buffered salt solution and incubated for 1 h at 37 degrees C in 300 microliter volumes in the absence or presence (9 X 10(-4) M) of calcium chloride. Addition of D-galactosamine hydrochloride (DGM; 2.8 X 10(-4)M) caused (in addition to basal release) a mean +/- SEM percent histamine release of 15.7 +/- 5.2 in the presence of Ca++ and 19 +/- 4.9 in the absence of Ca++ (p greater than 0.05). It is suggested that D-galactosamine does not require extracellular Ca++ for the release of histamine from the rat mast cell.     

Effect of periodate oxidation of glycoconjugates of lymphocyte membranes on the immune response in vitro]

Mouse spleen cells treated with sodium periodate for 10 min. at 4 degrees C are stimulated to undergo blastogenesis and to incorporate thymidine. The effect of such treatment on the antibody response in vitro induced by Sheep red blood cells has been evaluated. Periodate-induced proliferation is accompanied by a marked inhibition of the immune response to this antigen. At concentrations leading to mitogenesis, no cytotoxic effect of periodate was observed and treated cells survived well on tissue culture. Cell recoveries from samples treated with periodate at the optimal mitogenic dose, were markedly enhanced when harvested at different days after culturing wheras lower antibody forming cells numbers wereconsistently observed during the culture period.     

Dissociation-constants of metat-ion-complexes with alkaline phosphatase from pig kidney.

Using metal-ion buffers it was possible to remove Zn2+, Mg2+ and Mn2+ ions of pig kidney alkaline phosphatase reversibly. The dissociation constants obtained are KEMg: 4 X 10(-7) M, KEMn: 4 X 10(-8) M and KEZn: 8 X 10(-13) M (22 degrees C, pH: 9.6, mu: 0.07).     

Parameters affecting the maximum cell concentration of Tetrahymena

In cultures with efficient aeration a maximum cell concentration (MCC) of 6 X 10(5) cells/ml (defined medium) and 5.5 X 10(6) cells/ml (broth) can be reached. By culturing within Millicells with excess supply of medium and efficient removal of waste products a physical limit for MCC of about 13 X 10(6) cells/ml is reached.     

Diffusion loading conditions determine recovery of protein synthesis in electroporated P3X63Ag8 cells

Using the suspension cell line P3X63Ag8 we have studied the impact of the composition of the diffusion medium on cellular protein synthesis under standard electroporation conditions in TBS-Na. This buffer contains the high saline concentration usually present in electroporation-mediated DNA transfection. Electroporation in the presence of TBS-Na resulted in an immediate shut-off of protein synthesis, even though both FITC-dextran (Mr 40 kD) and Semliki Forest virus core protein (Mr 33 kD) were incorporated efficiently into the cytoplasm across the electropores at 0 degrees C. Subsequent resealing of the pores was completed after a 5-min incubation at 37 degrees C. When compared with control cells, overall protein synthesis of electroporated cells recovered slowly to resume a 30% activity after 1 h of incubation at 37 degrees C. We have determined optimal conditions for diffusion loading (which necessitates the presence of ATP, GTP, amino acids, K+, Mg2+, and Ca2+) and resealing (in the presence of K+, Mg2+, and Ca2+), leading to a full and lasting recovery of protein synthesis within 5 min after pore closure.     

Regulation of ammonia uptake in aspergillus nidulans.

The ammonia uptake in A. nidulans was found to be linear for about 20 min, and was proportional up to 1.5 mg/ml dry cell density. The transport of ammonia does not involve energy. Normal and biotin deficient A. nidulans showed an identical Km-values of 10.26 X 10(-5) M ammonia for uptake. The uptake of ammonium ion has been shown to be regulated by the intracellular concentration of ammonia.     

Role of calcium in the histamine release induced by D-galactosamine from rat mast cells

Summary Rat peritoneal mast cells were isolated and purified by differential centrifugation in Ficoll. Cells pooled from three to four rats were suspended at approximately 10⁶ cells/ml in a buffered salt solution and incubated for 1 h at 37°C in 300 l volumes in the absence or presence (9×10^–4 M) of calcium chloride. Addition of D-galactosamine hydrochloride (DGM; 2.8×10^–4 M) caused (in addition to basal release) a mean ±SEM percent histamine release of 15.7±5.2 in the presence of Ca⁺⁺ and 19±4.9 in the absence of Ca⁺⁺ (p>0.05). It is suggested that D-galactosamine does not require extracellular Ca⁺⁺ for the release of histamine from the rat mast cell.A preliminary analysis of these results was presented at the International Symposium on calcium entry blockers and tissue protection, Rome, 15–16 March 1984.     

Differences in luteinizing hormone release stimulated in rat anterior pituitary cells by leukotriene C4 and by gonadotropin-releasing hormone in vitro

Continuous administration of leukotriene C4 (LTC4, 10(-10) M) to superfused rat anterior pituitary cells increased LH release for 40 min only, whereas in a parallel experiment gonadotropin-releasing hormone (GnRH, 10(-9) M) evoked a continuous increase in hormone secretion. In contrast to GnRH, LTC4 did not desensitize rat anterior pituitary cells. The secretory action resulting from the administration of LTC4 (10(-10) M) was abolished for 40 min after previous stimulation. The results documented the dual action of LTC4 on LH exocytosis.     

Survival of rabbit embryos after rapid freezing and thawing

Summary Frozen storage of rabbit embryos at the 16-cell stage in 2.0 M dimethylsulfoxide (DMSO) in phosphate-buffered saline (PBS) was achieved by a 2-step procedure. After storage for 10 days at–196°C they were revived by rapidly thawing at 500°C/min. On transfer of these embryos to pseudopregnant foster mothers, 50% survived to term. The difference in in vivo survival between frozen-thawed and frozen-thawed-cultured embryos was not significant.Acknowledgment. The authors thank the Director for the facilities. VHR is supported by Council of Scientific and Industrial Research (India).     

Persistence of added retinoids in organ culture media during induction of mucous metaplasia and glandular morphogenesis in hamster cheek pouches

The retinoid concentration (determined colorimetrically) did not change significantly in retinyl acetate-supplemented (6 micrograms/ml) Eagle's Minimal Essential Medium containing 10% fetal calf serum when stored at -20 or 4 degrees C over 7 days. After the medium was incubated at 37 degrees C for 48 h, 37-49% of the retinoid remained, whether or not tissue (neonatal Syrian hamster cheek pouch) was present, and irrespective of explant age. The normal retinoid level in the tissue was approximately 0.25 micrograms per gram. Therefore, neonatal hamster cheek pouches, incubated in medium with the addition of 6 micrograms of retinyl acetate per ml of medium and undergoing mucous metaplasia and some mucous gland morphogenesis, were continually being exposed to retinoid levels which, though gradually decreasing, remained well above their normal physiological level.     

Is glucagon involved in cold acclimatization?

Cold acclimatization in rats at 5 degrees C for 2 weeks caused a significant elevation of plasma glucagon concentration, accompanied by increased plasma FFA and glucose levels. Acute cold exposure at 5 degrees C for 5 or 60 min did not affect these parameters in plasma.     

Improved separation at low temperature of glycoproteins by Con A-Sepharose affinity chromatography in the presence of sodium dodecyl sulfate (SDS)

The Con A-Sepharose affinity chromatography of glycoproteins was even more effective at 4 degrees C than that at room temperature (26-28 degrees C) in the presence of sodium dodecyl sulfate (SDS). Application of this methodology to the separation of several glycoproteins from SDS-solubilized membrane proteins in rat cerebellum, including a glycoprotein characteristic of the Purkinje cells, was successful.     

基于MATLAB的AA—HPA—AMPS—AM聚合物阻垢剂合成及其性能研究

通过溶液聚合方法,以过硫酸铵为引发剂,丙烯酸、丙烯酸羟丙酯、2-丙烯酰胺-2-甲基丙磺酸、丙烯酰胺为单体,合成AA-HPA-AMPS-AM聚合物.固定聚合物浓度为4 mg/L,分别考查反应时间、反应温度（X1）、引发剂用量（X2）、单体配比（AA：HPA以X3计、AMPS：AM以X4计）对聚合物阻CaCO3垢性能的影响,并设计了正交实验,得出当聚合物在80℃、引发剂用量为10%、M（AA）：M（HPA）为0.25、M（AMPS）：M（AM）为0.4时,阻CaCO3垢率最高为52.9%.为探究此聚合物是否有更高的阻垢性能,应用MATLAB对正交实验结果进行分析及验证,得知：聚合物阻CaCO3垢效率y与各影响因素X之间基本满足y=146.349 7-0.926 7x1-49.765 8x3-28.220 9x4.最佳聚合条件为x1=75、x3=0.15、X4=0.15,y为68.35%.     

Lack of participation of cytoplasmic choline in the synthesis of acetylcholine in the striatal synaptosomes of the rat]

Synaptosomes prepared from Rat striatum were loaded with (14C) choline at 0 degrees C in the course of a 60 min. preincubation. After the removal, by washing, of the extracellular molecules, synaptosomes were incubated 5 min. at 37 degrees C in the presence of (3H) choline. The measurement of the amounts of (3H) ACh and of (14C) ACh formed allowed us to conclude that the acetylation of the molecules of choline was directly coupled to their passage through the synaptosomal membrane.     

Effect of actinomycin D on Trypanosoma cruzi

Viable metacyclic forms of T. cruzi, Y strain, treated with an adequate dose of actinomycin D (50 micrograms Act-D/ml/10(7) parasites/ml for 72 h at 28 degrees C) showed the following properties: 1) they lost their ability to replicate in culture medium, in blood and in tissues of normal mice and were no longer able to incorporate tritiated thymidine; 2) they could not penetrate into Vero cells and could not replicate inside normal macrophages; 3) they retained their immunogenicity and the ability to protect mice against a virulent infection; 4) they did not induce histological lesions as described in chronic experimental Chagas' disease.     

The effects of time of equilibration with cryoprotectants at 0 degree C prior to freezing on the survival of mouse embryos frozen by the two-step method

Mouse embryos were frozen by the two-step method after equilibration for 0.1-60 min with cryoprotectants at 0 degree C. No survival or a very low survival was obtained after equilibration for only 0.1 min. The morulae showed the highest survival rates when equilibration time was 5-30 min with 2 M DMSO, 20-30 min with 2 M glycerol, 5-10 min with 2 M ethylene glycol and 20-30 min with 2 M propylene glycol, respectively.     

Inhibition of lymphocyte blastogenesis by diazepam, meprobamate, and other psychotropic drugs]

Imipramine, diazepam, chlordiazepoxide, and meprobamate inhibit incorporation of 3H-thymidine in human lymphocytes in culture. For the first three drugs, 50% inhibition occurs at a concentration of 5 X 10(-5) M, and for meprobamate at a concentration of 10(-3 M. Ethanol has no action up to 10(-1) M.     

The effects of time of equilibration with cryoprotectants at 0°C prior to freezing on the survival of mouse embryos frozen by the two-step method

Summary Mouse embryos were frozen by the two-step method after equilibration for 0.1–60 min with cryoprotectants at 0°C. No survival or a very low survival was obtained after equilibration for only 0.1 min. The morulae showed the highest survival rates when equilibration time was 5–30 min with 2 M DMSO, 20–30 min with 2 M glycerol, 5–10 min with 2 M ethylene glycol and 20–30 min with 2 M propylene glycol, respectively.     

Alteration in thermal stability of ribosomes from Drosophila melanogaster with age.

Thermal analysis of high salt (0.5 M) washed ribosomal monomers from young and old male Drosophila melanogaster revealed an 8 degrees C downshift in the mean temperature of denaturation (Tm). Moreover, there was observed a marked loss in the ability of ribosomes extracted from older flies to reassociate upon cooling. These observations suggest that age-dependent alterations in the structural integrity of the rRNA-r protein complex could, at least in part, be responsible for the diminished capacity for protein synthesis in this species with advancing age.     

Antithyroid effect of a food or drug preservative: 4-hydroxybenzoic acid methyl ester

Summary 4-hydroxybenzoic acid methyl ester (methylparaben) inhibits organification of iodide by isolated hog thyroid cells. The concentration which produces a 50% inhibition was about 2.0×10^–4M. A similar inhibition was observed in nonstimulated and TSH- or dibutyryl cyclic AMP-stimulated cells. Neither iodide uptake nor cyclic AMP generation were altered by methylparaben.