X-Ray radiography for studying diffusion characteristics of sucrose in plaster of paris matrix containing yeast cells

Summary X-Ray radiography was employed to monitor the diffusion of sucrose into plaster of Paris matrix containing 20% yeast cells. It was observed that the depth of penetration of tracer P_b detected by radiography matched with the substrate penetration detected by chemical test. However electron probe microanalysis (EPMA) did not yield any conclusive evidence regarding the movements of tracer P_b and substrate to the same extent.     

A high affinity Ca2(+)-ATPase on the surface membrane of Leishmania donovani promastigote

A Ca2(+)-dependent ATP-hydrolytic activity was detected in the crude membrane ghost of the promastigote or vector form of the protozoal parasite Leishmania donovani, the pathogen responsible for kala azar. The Ca2(+)-ATPase was purified to apparent homogeneity after solubilization with deoxycholate. The enzyme consists of two subunits of Mr = 51,000 and 57,000 and has an apparent molecular weight of 215,000 +/- 12,000. The enzyme activity is exclusively dependent on Ca2+, and the pure enzyme can hydrolyze 1.6 mumol of ATP/min/mg of protein. The apparent Km for Ca2+ is 35 nM, which is further reduced to 12 nM in the presence of heterologous calmodulin. The enzyme is sensitive to vanadate, but is insensitive to oligomycin and ouabain. The enzyme is strongly associated with the plasma membrane and has its catalytic site oriented toward the cytoplasmic face. The enzyme spans across the plasma membrane as surface labeling with radioiodine shows considerable radioactivity in the completely purified enzyme. The localization and orientation of this high affinity, calmodulin-sensitive Ca2(+)-ATPase suggest some role of this enzyme in Ca2+ movement in the life cycle of this protozoal parasite.     

Acid pH-induced fusion of cells by herpes simplex virus glycoproteins gB an gD

Enveloped animal viruses enter host cells either by direct fusion at neutral pH or by endocytosis. Herpes simplex virus (HSV) is believed to fuse with the plasma membrane of cells at neutral pH, and the glycoproteins gB and gD have been implicated in virus entry and cell fusion. Using cloned gB or gD genes, we show that cells expressing HSV-1 glycoproteins gB or gD can undergo fusion to form polykaryons by exposure only to acidic pH. The low pH-induced cell fusion was blocked in the presence of monoclonal antibodies specific to the glycoproteins. Infection of cells expressing gB or gD glycoproteins with HSV-1 inhibited the low pH-induced cell fusion. The results suggest that although the glycoproteins gB and gD possess fusogenic activity at acidic pH, other HSV proteins may regulate it such that in the virus-infected cell, this fusion activity is blocked.     

Shedding of vesicular stomatitis virus soluble glycoprotein by removal of carboxy-terminal peptide.

A comparison of partial NH2-terminal sequences of vesicular stomatitis viral glycoprotein G (molecular weight, 69,000) and the soluble extracellular glycoprotein antigen Gs (molecular weight, 57,000) shows that both of the sequences are identical. Tryptic fingerprint analyses show that Gs lacks the carboxy-terminal region containing the membrane-anchoring hydrophobic domain of G. These results suggest that Gs is formed by cleavage in the carboxy-terminal region of G.     

Biochemical mechanism of nitrofurantoin resistance inVibrio el tor

Vibrio el tor cells contain a constitutive reductase enzyme which converts nitrofurantoin to an active principle that is responsible for the observed antibacterial activity of the drug. Acquisition of resistance of this strain towards nitrofurantoin is associated with the loss of this reductase. This enzyme is located in the periplasmic region of the nitrofurantoin-sensitive cells, and seems to play an important role in transporting the drug into the cells.     

Assessment of radiosensitivity from the germination of jute seeds

Irradiation with X-rays and gamma rays reduced the speed of germination of seeds of jute cultivars,viz., JRO 632, JRO 620, Sudan Green ofCorchorus olitorius and JRC 212, Fanduk, D 154 ofC. capsularis. Though the first phase of germination(i.e. seed variability) remained apparently unaffected, the second phase (i.e. the sprouting ability) and the third phase(i.e. the attainment of autotrophic status) were found to be the best indicators of radiation injury and provided dependable data for the assessment of radiosensitivity. The jute cultivars have not shown accountable intervarietal differences in radiosensitivity in respect of LD 50 and LD 100 for the second and the third phases of germination.     

The characterisation and function of the polysaccharidases of human synovial fluid in rheumatoid and osteoarthritis.

A potential enzymic mechanism for the degradation of glycosaminogly cans was characterised using enzymes found in rheumatoid synovial fluid from the knee joint. This mechanism involves a true hyluronidase together with the concerted action of beta-glucuronidase and beta-N-acetylhexosaminidase. The contribution of the exopolysaccharidases to hyaluronate degradation was demonstrated by the use of specific inhibitors, while the distinct identity of a true hyaluronidase was shown by ammonium sulphate and agarose gel column fractionations. Only the hyluronidase fraction was capable of degrading high molecular weight hyaluronate. The exopolysaccharidase activities were shown to be markedly elevated in rheumatoid as compared to osteoarthritic synovial fluid and also normal serum. On the other hand, hyluronidase was similarly active in rheumatoid and osteoarthritic synovial fluids; both these levels were lower than that of normal human serum. Hyaluronidase in synovial fluid may thus be derived by diffusion from serum, since it is of relatively low molecular weight (60 000). The pH requirements of this enzyme system and the strong inhibition of hyaluronidase by synovial fluid make it unlikely that the mechanism operates extracellularly. It is proposed that as a lysosomal mechanism, however, it is an important contributing factor in the chronic erosion process characteristic of rheumatoid arthritis.     

Effect of 3-(3,4-dihydro-6-methoxy-2-naphthyl)2,2-dimethyl pentanoic acid--an antifertility agent--on in vitro uptake of estradiol 17beta-6, 7(3)-H in rat uterus.

The effect of 3-(3,4-dihydro-6-methoxy-2-naphthyl)-2,2-dimethyl-pentanoic acid, a potent nonsteroidal antifertility compound, on the uptake of estradiol-17beta-6, 7-tritiated in vitro in the rat uterus was studied. The estradiol uptake of estrogen-primed and compound-treated groups were the same. When estradiol and the compound were present in medium at the same time, estradiol uptake was significantly (p less than .01) increased. The results indicate that the compound synergizes the effect of estradiol.