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Biomedical subjects

J Das

Publications and source records attributed to J Das.

At least 127 records · Page 7Linked to original sources

Phosphate repression of phage protein synthesis during infection by choleraphage phi 149.

A synthetic medium for choleraphage phi 149 growth, in which the concentration of phosphate ions plays a significant role, has been defined. Upon infection, choleraphage phi 149 DNA binds to the cell membrane at three to four sites. The host macromolecular syntheses are shut off by 10 min after infection and the synthesis of phage-specific DNA is detectable after 20 min of infection. The phage utilizes primarily the host DNA degradation products for its own DNA synthesis. When added during the first 20 min of infection both nalidixic acid and novobiocin inhibit phage growth. The effects of these antibiotics are not pronounced when added late during infection. Pulse labeling of ultraviolet-irradiated infected cells at different times during infection has allowed identification of about 50 phage-specific proteins of which 19 are structural proteins. These proteins appear during the infection cycle in two distinct phases, early and late. When infection is carried out in high-phosphate medium, none of the late proteins is synthesized. Eleven of the 26 early proteins detected are DNA-binding proteins.

Bacteriophages↗

Lysis of Vibrio cholerae cells: direct isolation of the outer membrane from whole cells by treatment with urea.

Cells of Vibrio cholerae underwent rapid autolysis when suspended in media of low osmolarity under non-growing conditions. Chaotropes like urea and guanidine. HCl which are potent protein denaturants caused complete and immediate lysis of whole cells. This unique sensitivity of V. cholerae to protein denaturants led to the development of a rapid method for the selective isolation of the outer membrane upon treatment of whole cells with urea. The composition of the outer membrane isolated from both whole cells and crude envelopes by treatment with urea was comparable with that of the outer membrane isolated by other conventional methods.

Bacterial Outer Membrane Proteins↗

Continuous telemetric monitoring of bladder function.

Preliminary investigations with telemetric urodynamic equipment permit us to describe the following indications for this technique: (1) repeated pressure/flow studies, both in obstruction and in experimental work; (2) when urge incontinence is expected on the grounds of the patient's history but standard cystometry reveals a stable bladder; (3) enuresis nocturna and psychogenic voiding dysfunctions; (4) in the therapy of incontinence. Some examples are presented.

Adult↗

Radiation-sensitive mutant of hypertoxinogenic strain 569B of Vibrio cholerae.

A radiation-sensitive mutant of the hypertoxinogenic strain 569B of Vibrio cholerae was isolated and characterized. The mutant, designated V. cholerae 569Bs, lacks both excision- and medium-dependent dark-repair mechanisms of UV-induced DNA damage while retaining the wild-type photoreactivating capability. Analysis of the UV-irradiated cell DNA by velocity sedimentation in alkaline sucrose gradient suggests that UV-induced pyrimidine dimers may not be incised in these cells. In contrast to the wild-type cells, the mutant cell DNA was degraded after treatment with nalidixic acid. The mutant cells failed to produce any detectable amount of cholera toxin as measured by ileal-loop assay.

Bacteriophages↗

Repair of ultraviolet light-induced DNA damage in cholera bacteriophages.

DNA repair-proficient and -deficient strains of Vibrio cholerae were used to examine host cell reactivation, Weigle reactivation and photoreactivation of u.v.-irradiated cholera bacteriophages. U.v. light-induced DNA damage in phages of different morphological and serological groups could be efficiently photoreactivated. Host cell reactivation of irradiated phages of different groups was different on the same indicator host. Phage phi 149 was the most sensitive, and phi 138 the most resistant to u.v. irradiation. While phi 138 showed appreciable host cell reactivation, this was minimal for phi 149. Attempts to demonstrate Weigle reactivation of u.v.-irradiated cholera phages were not successful, although u.v.-induced filamentation of host cells was observed.

Bacteriophages↗

Repression of the alkaline phosphatase of Vibrio cholerae.

The synthesis of alkaline phosphatase by two strains of Vibrio cholerae belonging to the Inaba and Ogawa serotypes has been examined in relation to the phosphate concentration of the culture medium. The synthesis of the enzyme in both strains was repressed in cells grown in the presence of a high concentration of inorganic phosphate. Lowering the phosphate content of the growth medium led to a derepression of enzyme activity. The presence of glucose in low phosphate medium stimulated the degree of derepression. The synthesis of the enzyme by strain Inaba 569B was more sensitive to inorganic phosphate than that of strain Ogawa 154. The enzyme was presumably located in the periplasmic space since it was released when the organisms were converted to spheroplasts.

Alkaline Phosphatase↗

Monomeric alkaline phosphatase of Vibrio cholerae.

Alkaline phosphatase has been purified to homogeneity from two strains of Vibrio cholerae. The enzymes from both strains are single polypeptides of molecular weight 60,000. Both of the enzymes have pH optima around 8.0 and can act on a variety of organic phosphate esters, glucose-1-phosphate being the best substrate. The enzymes are unable to hydrolyze ATP and AMP. Although they have identical Km values, the two enzymes differ significantly in Vmax with p-nitrophenyl phosphate as substrate. The enzymes from the two strains also differ in their sensitivity to EDTA, Pi, and metal ions and activities of the apoenzymes. Ca2+ reactivated the apoenzymes most.

Alkaline Phosphatase↗

Replication of mycoplasma virus L51. VII. Effect of chloramphenicol on the synthesis of DNA replicative intermediates.

Chloramphenicol affects several steps in the DNA replication of mycoplasma virus L51, a noncytocidal, naked, bullet-shaped virion containing circular single-stranded (SS) DNA of 1.5 X 10(6) daltons (4.5 kilobases). In the presence of chloramphenicol, adsorption was normal and parental SS DNA was converted to double-stranded replicative forms (RF), but subsequent RF leads to RF replication was inhibited. Chloramphenicol added late in infection, when most viral nascent DNA is in progeny SS molecules, inhibited SS synthesis, but nascent RF molecules were formed. However, a chase experiment showed that these RF molecules could not be converted to SS DNA. Therefore, viral RF molecules made in the presence of chloramphenicol are not functional as SS DNA precursors.

Acholeplasma laidlawii↗

Repair of ultraviolet-light-induced DNA damage in vibrio cholerae.

Repair of ultraviolet-light-induced DNA damage in a highly pathogenic Gram-negative bacterium, Vibrio cholerae, has been examined. All three strains of V. cholerae belonging to two serotypes, Inaba and Ogawa, are very sensitive to ultraviolet irradiation, having inactivation cross-sections ranging from 0.18 to 0.24 m2/J. Although these cells are proficient in repairing the DNA damage by a photoreactivation mechanism, they do not possess efficient dark repair systems. The mild toxinogenic strain 154 of classical Vibrios presumably lacks any excision repair mechanism and studies of irradiated cell DNA indicate that the ultraviolet-induced pyrimidine dimers may not be excised. Ultraviolet-irradiated cells after saturation of dark repair can be further photoreactivated.

DNA Repair↗

Ultraviolet, photodynamic and thermal inactivation of mycoplasmaviruses.

The ultraviolet light, photodynamic, and thermal inactivation parameters have been measured for the three groups of mycoplasmaviruses. The differences in these parameters for the three virus groups reflect differences in virion structure and in genome size and structure.

Acholeplasma laidlawii↗

Photodynamic inactivation and its repair in mycoplasmas.

Photodynamic inactivation is the loss in viability observed when organic dye-treated cells are exposed to visible light and molecular oxygen. The photodynamic inactivation of mycoplasmas, the smallest free living cells, has been studied. Depending on the extent of inactivation in Acholeplasma laidlawii, photodynamic induced damage can be repaired if the irradiated cells are incubated in the dark in buffer. Analysis of the DNA of these cells shows that photodynamic inactivation induces single strand breaks which can be repaired during liquid holding. To examine possible damage to the cell membrane, glucose uptake was studied as a permeability measure. Neither acriflavine nor photodynamic inactivation had any measurable effect on membrane permeability.

Acholeplasma laidlawii↗

Effect of acriflavine on ultraviolet inactivation of Acholeplasma laidlawii.

An increased sensitivity to inactivation was observed when ultraviolet light-irradiated Acholeplasma laidlawii cells were plated on medium containing either acriflavine or chloramphenicol. Chloramphenicol reduced liquid holding recovery (dark repair) to about 10% of that in untreated irradiated cells. In acriflavine treated cells no dark repair could be observed and there was a progressive degradation of cell DNA during holding. While the primary effect of acriflavine may be to inhibit excision repair, since ultraviolet-irradiated Mycoplasma gallisepticum (cells which lack an excision repair mechanism) show a slight increase in inactivation when plated on medium containing acriflavine the dye must also have some other effects on ultraviolet repair processes. Acriflavine treatment of A. laidlawii cells before ultraviolet irradiation has a protective effect, as seen by an increased cell survival.

Acholeplasma laidlawii↗