Histamine secretion from permeabilized mast cells by calcium

A transient increase in the permeability of the mast cell membrane was caused by the exposure of the cells to low concentrations of saponin, 5 or 10 micrograms/ml. These concentrations had very little effect in the absence of calcium but caused 35 to 50% histamine release, having the character of a secretory response, when 0.25 mM or more calcium was added to the medium. The dose-response curve was steep between 25 microM and 250 microM calcium and tended to flatten with higher concentrations. The release was associated with a pronounced increase in calcium uptake, which was faster than the histamine release. The membrane changes were slight as indicated by only 7 to 12% leakage of lactate dehydrogenase and by the absence of any detectable change in the electron micrographs. The transient nature of the membrane change is shown by the following experiment. When the cells were first exposed to saponin in the absence of calcium, the amount of histamine released by the subsequent incubation with calcium varied inversely with the time interval that elapsed before calcium was added. If calcium was added after 15 minutes no histamine release occurred. When calcium uptake was studied in the same manner, the stimulation of calcium uptake in saponin-treated cells also declined progressively with increasing intervals after the exposure to saponin when calcium was added. Stimulation of both histamine release and calcium uptake was inhibited by antimycin A, the inhibition curves with 10(-9)M to 10(-7)M antimycin A being similar. The effect on the calcium uptake by itself could explain the inhibition of histamine release. But the release was also inhibited by the calmodulin antagonists, W-7 and mepacrine, suggesting that the influx of calcium in the permeabilized cells acts primarily through calmodulin-mediated enzyme activation.     

Synthesis and biological evaluation of longanlactone analogues as neurotrophic agents

Longanlactone analogues were synthesized using a route featuring Friedel-Crafts acylation, Sonogashira coupling and 1,3-dipolar cycloaddition reactions. Structure–activity relationships were investigated for neurotrophic activity. Compound 6 was found to have the most potent neurotrophic activity among all the synthesized analogues in Neuro2a cells as evidenced by a battery of in vitro/cell based assays for assessment of neurogenic and potential neurotrophic activity including neurite outgrowth assay and real time PCR for popular markers of augmented neurotrophic activity. Compound 6 might serve as a template for further development of highly effective neurotrophic molecules.     

P190-B, a Rho-GTPase-activating protein, is differentially expressed in terminal end buds and breast cancer.

Microdissection and differential display PCR were used to identify genes preferentially expressed in the highly proliferative terminal end buds (TEBs) in the mammary gland of 45-day-old virgin rats. One clone exhibited 87% homology to the human p190-B gene encoding a novel Rho-Gap. Using in situ hybridization, p190-B was detected in both the TEBs and the terminal ducts, with the highest expression observed in the outer layer of TEBs. During normal mammary gland development, p190-B mRNA expression was highest in the virgin mammary gland and decreased during late pregnancy and lactation. Interestingly, increased levels of p190-B mRNA relative to the normal mammary gland were seen in a subset of murine mammary tumors that appeared to be less well differentiated and potentially more aggressive. Transient transfection of a p190-B expression construct into MCF-10A human mammary epithelial cells resulted in disruption of the actin cytoskeleton, which suggests a role for p190-B in regulating the signaling pathways that influence cell migration and invasion. These results suggest that p190-B may be required for virgin mammary gland development, and its aberrant expression may occur in breast cancer.     

Structural and mass spectral studies on some multiply bonded diosmium tetracarboxylate compounds

The compounds Os₂(O₂CCH₃)₄Cl₂,1 and Os(O₂CC₂H₅)₄Cl₂, 2, have been structurally characterized. Both compounds crystallize in space group P2₁/n. For 1 the unit cell parameters are a = 6.546(1) Å, b = 8.950(1) Å, c = 12.533(1) Å, β = 90.17(1)° and Z = 2; for 2 they are a = 6.792(2) Å, b = 10.519(3) Å, c = 13.372(4) Å, β = 89.27(3)° and Z = 2. Both molecules have approximate D_4th symmetry with important dimensions as follows: for 1, Os≡Os 2.314(1) Å, OsCl 2.448(2) Å; for 2, Os≡Os 2.316(2) Å, OsCl 2.430(5) Å. The mass spectra of these compounds as well as that of Os₂(O₂CC₃H₇)₄Cl₂,3, are reported and discussed.     

The anthocyanin pigmentation pattern in Hibiscus sabdariffa L. and its mode of inheritance,with special reference to the variation intermedius

The anthocyanin pigmentation patterns of stem, epicalyx, calyx, fading corolla, colour of anthers and petal spot character were studied in Hibiscus sabdariffa L.The stem and epicalyx pigmentation are controlled by the c and r genes. The two genes C and R are involved so far as calyx pigmentation, fading corolla colour and the anther and spot colours are concerned but an additional factor f is assumed to explain the segregation of the calyx pigmentation and fading corolla colour. Thus, a total of three genes (c, r. f) is distinguished, segregating independently.The intermedius type resembles the green-light-red rype of altissima in stem and epicalyx pigmentation but shows distinct differences in calyx pigmentation and fading corolla colour.     

Indian aconites

Of the two dozen species of aconite native to India, thirteen are known to possess drug value. Some of the latter are in commercial trade and offer valuable substitutes for the more commonly used species of Europe. Their nomenclature is revised in this article, with proper consideration given to their habitats and chemical constituents in order to facilitate identification and standardization of this drug.     

Morphological changes in an acidophilic bacterium induced by heavy metals

The Acidiphilium strains inhabit acidic mine regions where they are subjected to occasional environmental stresses such as high and low temperatures, exposure to various heavy metals, etc. Change in morphology is one of the strategies that bacteria adopt to cope with environmental stresses; however, no study on this aspect has been reported in the case of Acidiphilium sp. This work is an attempt using the acidophilic heterotrophic bacterium Acidiphilium symbioticum H8. It was observed that the maximum alterations in size occurred when the bacterium was exposed to sub-inhibitory concentrations of Cu and Cd. Loosely packed coccobacillus-type normal cells formed characteristic chains of coccoidal lenticular shape with constrictions at the junctions between them in the presence of Cd; Cu induced transformation of cells to become round shaped; Ni caused the cells to aggregate, but Zn showed no effect. Respective metal depositions on the cell surface were confirmed by scanning electron microscopy equipped with energy dispersive X-ray analysis. Cell bound Ca²⁺ ions were replaced by these metal ions and measured by inductively coupled plasma mass spectrometry from the culture filtrate. Cell shape changed only after the addition of sub-inhibitory concentrations of the metals, but in growth inhibitory concentrations it was similar to the normal cells.     

Systematic analysis of the effect of multiple templates on the accuracy of comparative models of protein structure

Background  

Although multiple templates are frequently used in comparative modeling, the effect of inclusion of additional template(s) on model accuracy (when compared to that of corresponding single-template based models) is not clear. To address this, we systematically analyze two-template models, the simplest case of multiple-template modeling. For an existing target-template pair (single-template modeling), a two-template based model of the target sequence is constructed by including an additional template without changing the original alignment to measure the effect of the second template on model accuracy.