Cell-free extract(s) of Pseudomonas putida catalyzes the conversion of cyanides,cyanates, thiocyanates,formamide, and cyanide-containing mine waters into ammonia

 Our isolate, Pseudomonas putida, is known to be capable of utilizing cyanides as the sole source of carbon (C) and nitrogen (N) both in the form of free cells and cells immobilized in calcium alginate. In the present study, the cell-free extract(s) were prepared from the cells of P. putida grown in the presence of sodium cyanide. The ability of enzyme(s) to convert cyanides, cyanates, thiocyanates, formamide and cyanide-containing mine waters into ammonia (NH₃) was studied at pH 7.5 and pH 9.5. The kinetic analysis of cyanide and formamide conversion into NH₃ at pH 7.5 and pH 9.5 by the cell-free extract(s) of P. putida was also studied. The K _m and V _max values for cyanide/formamide were found to be 4.3/8 mM and 142/227 μmol NH₃ released mg protein^-1 min^-1 respectively at pH 7.5 and 5/16.67 mM and 181/434 μmol NH₃ released mg protein^-1 h^-1 respectively at pH 9.5. The study thus concludes that the cell-free extract(s) of P. putida is able to metabolize not only cyanides, cyanates, thiocyanates, and formamide but also cyanide-containing mine waters to NH₃. Received: 10 April 1995/Received revision: 24 July 1995/Accepted: 22 August 1995     

Physiological causes of cotton fruit abscission under conditions of high temperature and enhanced ultraviolet-B radiation

An experiment was conducted in sunlit controlled environment growth chambers to determine the physiological mechanisms of fruit abscission of cotton ( Gossypium hirsutum L. cv. NuCOTN 33B) grown in high temperature and enhanced ultraviolet (UV)-B radiation. Six treatments included two levels of optimum (30/22°C) and high (36/28°C) day/night temperatures and three levels of biologically effective UV-B radiation (0, 7, and 14 kJ m⁻² per day). Both the temperature and UV-B treatments were imposed from seedling emergence through 79 days after emergence (DAE). High temperature did not negatively affect either leaf net photosynthetic rates (Pn) or abscission of cotton squares (floral buds with bracts) but significantly decreased boll retention. Plants exposed to 7 kJ UV-B radiation retained 56% less bolls than the 0 kJ UV-B control plants at 79 DAE, despite no significant differences in leaf Pn measured at squaring and flowering. At 53 DAE, leaf Pn of plants grown in high UV-B radiation (14 kJ m⁻² per day) decreased by 11%, whereas total non-structural carbohydrate (TNC) concentrations in the leaves, floral buds, and young bolls decreased by 34, 32, and 20%, respectively, compared with the control plants. The high UV-B radiation significantly increased square abscission. Square abscission was not related to leaf TNC concentration but closely correlated with TNC in floral buds ( r  = −0.68, P  < 0.001). Young boll abscission was highly correlated with TNC concentrations in both the leaves ( r  = −0.40, P  < 0.01) and the bolls ( r  = −0.80, P  < 0.001). Our results indicate that non-structural carbohydrate limitation in reproductive parts was a major factor associated with fruit abscission of cotton grown under high temperature and enhanced UV-B radiation conditions.     

Synthesis of thiazolidine-2,4-dione derivatives: anticancer,antimicrobial and DNA cleavage studies

In the search of efficient anticancer agents, here, new 5-(4-alkylbenzyledene)thiazolidine-2,4-dione derivatives (5a–g) have been successfully synthesized and characterized and are evaluated for anticancer and antimicrobial activities using DNA cleavage studies. In vitro studies on anticancer activity of compound 5d (NSC: 768619/1) was done against the full panel of 60 human tumor cell lines. The five-level dose activity results revealed that, the compound 5d was active against all the cell lines, it has shown potential activity against leukemia SR (GI₅₀: 2.04 μM), non-small cell lung cancer NCI-H522 (GI₅₀: 1.36 μM), colon cancer COLO 205 (GI₅₀: 1.64 μM), CNS cancer SF-539 (GI₅₀: 1.87 μM), melanoma SK-MEL-2 (GI₅₀: 1.64 μM), ovarian cancer OVCAR-3 (GI₅₀: 1.87 μM), renal cancer RXF 393 (GI₅₀: 1.15 μM), prostate cancer PC-3 (GI₅₀: 1.90 μM), and breast cancer MDA-MB-468(GI₅₀: 1.11 μM). DNA cleavage studies revealed that at 50 μg/mL concentration, partial DNA digestion was observed and when the concentration is increasing to threefold (150 μg/mL), complete linear DNA digestion and partial supercoiled DNA digestion was observed. Further antimicrobial studies indicate that all the synthesized compounds except compound 5a possess prominent activity against all the screened microbial species. This study throws a ray of light in the field of anticancer drugs.     

Impacts of management practices on bioenergy feedstock yield and economic feasibility on Conservation Reserve Program grasslands

Perennial grass mixtures planted on Conservation Reserve Program (CRP) land are a potential source of dedicated bioenergy feedstock. Long‐term nitrogen (N) and harvest management are critical factors for maximizing biomass yield while maintaining the longevity of grass stands. A six‐year farm‐scale study was conducted to understand the impact of weather variability on biomass yield, determine optimal N fertilization and harvest timing management practices for sustainable biomass production, and estimate economic viability at six CRP sites in the United States. Precipitation during the growing season was a critical factor for annual biomass production across all regions, and annual biomass production was severely reduced when growing season precipitation was below 50% of average. The N rate of 112 kg ha^?1 produced the highest biomass yield at each location. Harvest timing resulting in the highest biomass yield was site‐specific and was a factor of predominant grass type, seasonal precipitation, and the number of harvests taken per year. The use of N fertilizer for yield enhancement unambiguously increased the cost of biomass regardless of the harvest timing for all six sites. The breakeven price of biomass at the farmgate ranged from $37 to $311 Mg^?1 depending on the rate of N application, timing of harvesting, and location when foregone opportunity costs were not considered. Breakeven prices ranged from $69 to $526 Mg^?1 when the loss of CRP land rental payments was included as an opportunity cost. Annual cost of the CRP to the federal government could be reduced by over 8% in the states included in this study; however, this would require the biomass price to be much higher than in the case where the landowner receives the CRP land rent. This field research demonstrated the importance of long‐term, farm‐scale research for accurate estimation of biomass feedstock production and economic viability from perennial grasslands.     

Fluorodensitometric evaluation of gentamicin from plasma and urine by high-performance thin-layer chromatography

High-performance thin-layer chromatographic (HPTLC) analysis of gentamicin by in situ fluorodensitometric evaluation of its 4-chloro-7-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl) derivative is presented. The aminoglycoside components separated on silica gel plates using chloroform–methanol–20% ammonium hydroxide (2.4:2.2:1.5, v/v/v) as the mobile phase were reacted with NB-Cl to yield highly fluorescent derivatives. The calibration curves of gentamicin in water, plasma and urine were linear in the range 40–200 ng. The mean values of intercept, slope and correlation coefficient were 16.82±0.473, 6.83±0.015 and 0.9968±0.0017 for standard curves in water, 17.35±0.375, 6.85±0.018 and 0.9941±0.0012 for standard curves in plasma and 14.35±0.286, 6.86±0.002 and 0.9933+0.0011 for standard curves in urine respectively. The analytical technique was validated for within-day and day-to-day variation. The results indicate that HPTLC, coupled with in situ fluorodensitometry, is a reliable and valuable technique for quantitative analysis of the bulk drug gentamicin and gentamicin from urine and plasma.     

Freeze Thaw: A Simple Approach for Prediction of Optimal Cryoprotectant for Freeze Drying

The present study evaluates freeze thaw as a simple approach for screening the most appropriate cryoprotectant. Freeze–thaw study is based on the principle that an excipient, which protects nanoparticles during the first step of freezing, is likely to be an effective cryoprotectant. Nanoparticles of rifampicin with high entrapment efficiency were prepared by the emulsion-solvent diffusion method using dioctyl sodium sulfosuccinate (AOT) as complexing agent and Gantrez AN-119 as polymer. Freeze–thaw study was carried out using trehalose and fructose as cryoprotectants. The concentration of cryoprotectant, concentration of nanoparticles in the dispersion, and the freezing temperature were varied during the freeze–thaw study. Cryoprotection increased with increase in cryoprotectant concentration. Further, trehalose was superior to fructose at equivalent concentrations and moreover permitted use of more concentrated nanosuspensions for freeze drying. Freezing temperature did not influence the freeze–thaw study. Freeze-dried nanoparticles revealed good redispersibility with a size increase that correlated well with the freeze–thaw study at 20% w/v trehalose and fructose. Transmission electron microscopy revealed round particles with a size ∼400 nm, which correlated with photon correlation spectroscopic measurements. Differential scanning calorimetry and X-ray diffraction suggested amorphization of rifampicin. Fourier transfer infrared spectroscopy could not confirm interaction of drug with AOT. Nanoparticles exhibited sustained release of rifampicin, which followed diffusion kinetics. Nanoparticles of rifampicin were found to be stable for 12 months. The good correlation between freeze thaw and freeze drying suggests freeze–thaw study as a simple and quick approach for screening optimal cryoprotectant for freeze drying.