Forward mutation assay of V79 cells to 6-thioguanine resistance in a soft agar technique that eliminates effects of metabolic co-operation

A technique involving culture in soft agar was used for the assay of forward mutation of V79 cells to 6-thioguanine (6TG) resistance. The main reason for the use of soft agar was to prevent reduction in recovery of mutants depending on the cell density plated for mutation selection, which is the chief problem in the liquid method, and which results mainly from metabolic co-operation due to cell-to-cell contact.V79 cells grew well in fortified soft agar medium (DMEM + 20% FBS) showing cloning efficiencies (>80%) as high as in liquid culture. Therefore, V79/HGPRT mutagenesis could be assayed quantitatively in soft agar culture.The frequency of 6TG-resistant colonies in agar selective medium increased linearly with increase in concentration of EMS. Toxicity and mutagenic responses were greater in soft agar than in liquid culture.In cultures of untreated and EMS-treated cells, more than 95% of the 6TG-resistant colonies isolated were aminopterin-sensitive.Use of soft agar for selection prevented the reduction in the number of mutants with increase in the size of incula on plating up to 1?2 × 10⁶ cells per 9-cm dish: in liquid culture, even with a lower plating number (2 × 10⁵ cells per 9-cm dish), a notable reduction in numbers of mutants was observed. This character was re-examined in a reconstruction experiment. The results show that, when up to 2 × 10⁶ cells were plated per 9-cm dish, 6TG-resistant cells were almost completely recovered from the soft agar medium, whereas only 10% were recovered from liquid culture.     

Mechanisms of plasmid-mediated antibiotic resistances in Vibrio parahaemolyticus

The mechanisms of drug resistance of clinical isolate, Vibrio (V.) parahaemolyticus ST550, resistant to chloramphenicol (CP), aminoglycoside antibiotics (AGs) and beta-lactam antibiotics were investigated. The mechanisms of resistance to CP, AGs and beta-lactam antibiotics were dependent on chloramphenicol acetyltransferase (CAT), aminoglycoside-3"-adenylyltransferase AAD(3") and aminoglycoside-3'-phosphotransferase APH(3') and TEM type penicillinase, respectively.     

Does the seston of Lake Biwa release dissolved inorganic nitrogen and phosphorus during aerobic decomposition?: Its implication for eutrophication

Seston was collected from depths of 5 and 60 m in the north basin and a depth of 0 m in the south basin of Lake Biwa in the summer of 1985 and decomposed in the laboratory under aerobic conditions for three months. There was no net release of dissolved inorganic phosphorus from any seston, in contrast with the liberation of dissolved inorganic nitrogen. From this result and other available data, it is suggested that almost all phosphorus entering Lake Biwa and remaining within it is finally transformed into particulate phosphorous, transported to the bottom sediments, and fixed there without being recycled. Thus, this mechanism seems to play an important role in the prevention of rapid eutrophication of Lake Biwa, in spite of high external phosphorus loading.     

Caprolactam,a light-promoted growth inhibitor in sunflower seedlings

A neutral growth inhibitor, isolated from methanolic extracts of sunflower seedlings, was characterized by spectral data as caprolactam. Light-grown se     

Value of chemical mixture in multiple supralethal X-irradiation of mice

A chemical mixture containing 25 Amoles of MEA, 7 pmoles of AET, and 2 micromoles of 5-HT was found to be of significant value in protecting mice against repeated exposures of 800 R, 1100 R, or 1400 R given at intervals of 28 days. Dose-reduction factors of 2.17, 2.18, 1.95, 2.14, and 1.58 were obtained for the first five exposures. Following the first exposure there was no chemical mortality. The beneficial value of this mixture, however, was limited by the incidence of chemical toxicity which was more prevalent in mice with higher cumulative doses of radiation.