Changes taking place in the electrophoretic haemolymph protein pattern during metamorphosis of Polytela gloriosae (Lepidoptera)

The haemolymph proteins of the larva, pupa and adult of Polytela gloriosae have been fractioned by Polyacrylamide gel disc electrophoresis. In the haemolymph of the fifth instar larval stage a total of ten protein fractions have been detected. The concentration of the protein fractions 2, 3, 4, 9 and 10 shows oscillations in their concentration in the early fifth instar, middle fifth instar and late fifth instar larval stage. In all 11 protein fractionswere detected in the haemolymph of different stages of the pupa. The protein bands 1, 7 and 10 of the pupa appear newly in the haemolymph as these bands were not found in the haemolymph of the larvae. The protein fraction 9 of larva was not found in the pupa. In the haemolymph of adult insect sexual difference was observed in the haemolymph protein pattern. In the haemolymph of adult female a total of 10 protein fractions were detected while from the male haemolymph a total of 8 protein fractions were detected. The pupal band 7 was not found in the adults of both the sexes. In the haemolymph of larva and adult one pigmented protein fraction was observed. No pigmented protein fraction was found in the haemolymph of pupa. Iron - containing protein fraction and the acid mucopolysaccharides were not found in the haemolymph. The protein fractions 3, 4, 5, 6 and 7 of adult haemolymph were darkly stained by the Schiff reagent and, thus, they are the fractions of glycoprotein. One protein fraction of lipoprotein was also found in the haemolymph.     

Evidence for variation in the number of functional gene copies at the AmaR locus in chinese hamster cell lines

The hypothesis of functional hemizygosity has been examined for the α-amanitin resistant (Ama^R, a codominant marker) locus in a series of Chinese hamster cell lines. Ama^R mutants were obtained from different cell lines, e.g., CHO, CHW, M3-1 and CHO-Kl, at similar frequencies. After fractionation of different RNA polymerase activities in the extracts by chromatographic procedures, the sensitivity of the mutant RNA polymerase II towards α-amanitin was determined. While all of the RNA polymerase II activity in mutant CHO and CHO-Kl lines became resistant to α-amanitin inhibition, only about 50% of the activity is highly resistant in Ama^R mutants of CHW and M3-1 cell lines. The remaining activity in the latter cell lines shows α-amanitin sensitivity similar to that seen with the wild-type enzyme. This behaviour is similar to that observed with a 1:1 mixture of resistant and sensitive enzymes from CHO cells. These results, therefore, strongly indicate that while only one functional copy of the gene affected by α-amanitin is present in CHO and CHO-Kl cells, two copies of this gene are functional in the CHW and M3-1 cell lines.     

Evidence for functional hemizygosity at the Emtr locus in CHO cells through segregation analysis

The hypothesis of functional hemizygosity at the emetine-resistant (Emt^r, a non-X-linked recessive marker) locus in Chinese hamster ovary (CHO) cells has been examined by segregation analysis. The frequencies and the rates of segregation of the Emt^r and Thg^r (thioguanine-resistant, an X-linked recessive mutation) markers were determined from hybrids constructed between an Emt^r-Thg^r CHO cell line and various other Chinese hamster lines (V79, M3-1, CHO, GM7S, CHW and CHL). Thg^r segregants were obtained at similar frequencies (10^?2–10^?3) from all the hybrids. The frequency of segregation of the Emt^r marker, however, was similar to that of Thg^r only in the CHO × CHO hybrids and was much lower (10^?4–10^?6) than the CHO × other Chinese hamster hybrids. Similar results were obtained when the segregation rates for the two markers from various hybrids were determined. These results are consistent with the hypothesis that in CHO cells, the gene responsible for Emt^r is present in a single (functional) copy, whereas two copies of this gene are present in other Chinese hamster lines examined.     

Cellular reaction to group A beta-haemolytic streptococcal membrane antigen and its relation to complement levels in patients with rheumatic heart disease.

Cell-mediated immunity and blood complement activities were studied in 35 patients with chronic rheumatic heart disease (RHD) and 17 normal subjects. The T-cell population in patients with RHD was reduced, as were the CH50 and C3 complement levels. The response to phytohaemagglutinin stimulation was deficient, but the lymphocytes of patients with RHD showed increased avidity for 3H-thymidine when stimulated with specific streptococcal membrane antigen. No differences were found between patients with acute rheumatic activity and those without such activity. The susceptibility of individual patients may be related to the specific sensitisation of lymphocytes, while the fact that this persisted even when T-cell numbers had returned to normal may account for the well-known recrudescenses after streptococcal infections in these patients.     

The role of EGF in testosterone-induced reproductive tract differentiation

EGF is known to modulate a variety of cellular functions including differentiation. The aim of this investigation was to determine the role of EGF in androgen-induced masculine differentiation. Accordingly, a series of experiments were designed and the results are summarized as described below. (1) We found that the specific deprivation of EGF using anti-EGF serum during the period of masculine differentiation in an organ culture bioassay system resulted in the disintegration of the Wolffian system in a dose-dependent manner. (2) Exogenous EGF supplemented in the above experiment corrected the anti-EGF effect, suggesting a specific role of EGF. (3) Anti-EGF serum was also found to disrupt the differentiation even in the presence of exogenous testosterone, suggesting an effect independent of testosterone synthesis. (4) EGF was found to have a direct masculinizing effect both in vivo and in vitro; however, it was not able to mimic all masculinizing effects of testosterone. The mesonephric segment of the Wolffian duct was retained by EGF in the female fetal tract under in vitro conditions, and under in vivo conditions EGF was able to increase anogenital distance and to induce epididymis in some female fetal mice. (5) We were able to detect an EGF-like material in the fetal genital tract during differentiation and found that the level of this material increased with advancement of differentiation. Thus, it appears from the above results that EGF plays a role in testosterone-induced reproductive tract differentiation.     

Antiulcerogenic and anti-inflammatory effects of emodin, isolated from Rhamnus triquerta wall

Emodin, an anthraquinone derivative, isolated from the whole plant of R. triquerta, in 15 mg/kg dose (ip) exhibited anti-inflammatory activity against carrageenin-induced pedal inflammation in rats. In the same dosage it also showed antiulcer activity against 4 hr pylorus-ligated, aspirin and immobilization stress-induced gastric ulcers in rats. It decreased acid and pepsin output and augmented mucus secretion in terms of total carbohydrate: protein ratio in the gastric juice of aspirin treated pylorus-ligated rats, indicating that the antiulcerogenic effect of emodin may be due to its effect on gastric secretion.     

Efficacy of methyl 5(6)-(alpha-hydroxyphenylmethyl) benzimidazole-2-carbamate, a metabolite of mebendazole, against developing forms of experimental helminth parasites.

Methyl 5(6)-(alpha-hydroxyphenylmethyl) benzimidazole-2- carbamate, a metabolite of mebendazole, was evaluated against metamorphic forms of Ancylostoma ceylanicum in hamsters, Nippostrongylus brasiliensis in rats and cysticercoids of Hymenolepis nana in grain beetles. The test compound offered better action than mebendazole except against H. nana cysticercoids where the activity of the compound and mebendazole was comparable, but was inferior to the standard cestodicidal drug, praziquantel. The results suggest that the action was better by ip route compared to per os route of drug administration.     

Subcellular Localization, Partial Purification, and Characterization of a Dynorphin Processing Endopeptidase from Bovine Pituitary

An enzyme capable of cleaving dynorphin B-29 to dynorphin B-13 is present in bovine pituitary, with 40- to 50-fold higher specific activity in the posterior and intermediate lobes than in the anterior lobe. Subcellular fractionation of bovine neurointermediate pituitary shows that this enzyme is present in the peptide-containing secretory vesicles. The enzyme has been purified 2,800-fold from whole bovine pituitaries using ion-exchange and gel filtration chromatography. Purified dynorphin-converting enzyme has a neutral pH optimum, and is subsantially inhibited by the thiol-protease inhibitor p-chloromercuriphenylsulfonic acid, but not by serine or metalloprotease inhibitors. The purified enzyme processes dynorphin B-29 at Arg14, producing both dynorphin B-14 and dynorphin B-13 in a 5:1 ratio. No other cleavages are observed, suggesting that the activity is free from other proteases and is specific for single Arg sequences. Purified enzyme also processes dynorphin A-17 at the single Arg cleavage site, generating both dynorphin A-8 and A-9 in a 7:1 ratio. The tissue distribution, subcellular localization, and substrate specificity of this enzyme are consistent with a physiological role in the processing of dynorphin B-29 and dynorphin A-17, and possibly other peptides, at single Arg residues.     

Modulatory effects of metanil yellow on immunity in rodents.

Pathomorphological and immunological studies were carried out on rodents following oral administration of 0, 0.1, 0.25 and 0.5% (w/w) metanil yellow, mixed in diet, for 30 days. No significant change in hematologic parameters and histologic architecture of liver, kidney, mesenteric lymph node, thymus and urinary bladder was observed except for mild desquamation of intestinal villi and moderate changes in Peyer's patches of small intestine with higher doses. Among immunological parameters, significant enhancement in the primary humoral immune response (anti-SRBC IgM plaque forming cells of spleen) was observed with the lowest dose of metanil yellow while higher doses produced opposing effects. An elevated cutaneous delayed type hypersensitivity (DTH) reaction to SRBC was seen in 0.1% metanil yellow treated animals but higher doses did not influence the reaction. The treatment also caused changes in functional capabilities of macrophages. Although these immune alterations could hardly influence the local immunity of gut, as measured by the capacity of animals to cause rejection of Nippostrongylus brasiliensis parasite, the potential to modulate the immunity in general by metanil yellow however assumes considerable biological significance.