Introgression of Pi2 and Pi5 Genes for Blast (Magnaporthe oryzae) Resistance in Rice and Field Evaluation of Introgression Lines for Resistance and Yield Traits

Blast caused by Magnaporthe oryzae is the most devastating disease causing significant loss in rice production. The destructive nature of the disease is mainly due to the genetic plasticity of M. oryzae which complicates the breeding strategies. Blast can be effectively managed by the deployment of R genes. In this study, broad‐spectrum blast resistance genes Pi2 and Pi5 were introgressed independently into popular but blast susceptible rice variety, Samba Mahsuri (BPT5204) by applying marker‐assisted backcross breeding approach. Tightly linked markers AP5930 for Pi2 and 40N23r for Pi5 gene were used in foreground selection. Background selection helped to identify the lines with maximum recovery of recurrent parent genome (RPG). The RPG recovery in Pi2 introgression lines was up to 90.17 and 91.46% in Pi5 lines. Homozygous introgression lines in BC₃F₄ generation carrying Pi2 and Pi5 gene were field evaluated for blast resistance, yield per se and yield‐related traits. The lines showed resistance to leaf and neck blast in multilocation field evaluation. Improved BPT5204 lines with improvement for blast resistance were on par with original BPT5204 in terms of grain yield and grain features.     

Synthesis and in vitro microbiological evaluation of novel diethyl 6,6'-(1,4-phenylene)bis(4-aryl-2-oxo-cyclohex-3-enecarboxylates)

Novel bis cyclohexenone ester derivatives 14-19 were synthesized and characterized by their spectral data. In vitro microbiological evaluations were carried out for all the novel compounds 14-19 against clinically isolated bacterial and fungal strains. Compounds 15, 16, 18 against Staphylococcus aureus, 14, 15 against β-Haemolytic streptococcus, 15, 19 against Micrococcus luteus, 17, 18 against Salmonella typhii, 14, 17 against Shigella flexneri, 15 against Escherichia coli, 16 against Pseudomonas aeruginosa, 15, 18, 19 against Klebsiella pneumonia exhibited potent antibacterial activity at an minimum inhibitory concentration (MIC) value of 6.25 μg/ml, whereas compound 16 against Aspergillus flavus, 17 against A. niger, 16, 18 against Mucor indicus, 15, 17-19 against Microsporum gypseum revealed excellent antifungal activity at an MIC value of 6.25 μg/ml.     

A simple mortality risk prediction score for viper envenoming in India (VENOMS): A model development and validation study

BackgroundSnakebite is a neglected problem with a high mortality in India. There are no simple clinical prognostic tools which can predict mortality in viper envenomings. We aimed to develop and validate a mortality-risk prediction score for patients of viper envenoming from Southern India.MethodsWe used clinical predictors from a prospective cohort of 248 patients with syndromic diagnosis of viper envenoming and had a positive 20-minute whole blood clotting test (WBCT 20) from a tertiary-care hospital in Puducherry, India. We applied multivariable logistic regression with backward elimination approach. External validation of this score was done among 140 patients from the same centre and its performance was assessed with concordance statistic and calibration plots.FindingsThe final model termed VENOMS from the term “Viper ENvenOming Mortality Score included 7 admission clinical parameters (recorded in the first 48 hours after bite): presence of overt bleeding manifestations, presence of capillary leak syndrome, haemoglobin <10 g/dL, bite to antivenom administration time > 6.5 h, systolic blood pressure < 100 mm Hg, urine output <20 mL/h in 24 h and female gender. The lowest possible VENOMS score of 0 predicted an in-hospital mortality risk of 0.06% while highest score of 12 predicted a mortality of 99.1%. The model had a concordance statistic of 0·86 (95% CI 0·79–0·94) in the validation cohort. Calibration plots indicated good agreement of predicted and observed outcomes.ConclusionsThe VENOMS score is a good predictor of the mortality in viper envenoming in southern India where Russell’s viper envenoming burden is high. The score may have potential applications in triaging patients and guiding management after further validation.     

Effect of patulin on albumin fraction of plasma proteins studied in rats.

The toxic nature of the secondary metabolite of Penicillium patulum has been studied in rats. Plasma of the experimental animals showed a decrease in protein concentration. Detailed electrophoretic studies have been carried out, to find which fraction of the plasma protein is affected. It shows clearly that albumin fraction is very much affected, while in the tissues of the liver, kidney and intestine the DNA and RNA levels are found to be increased.     

Foot forces during typical days on the international space station

Decreased bone mineral density (BMD) in astronauts returning from long-duration spaceflight missions has been well documented, but the altered mechanical loading environment experienced by the musculoskeletal system, which may contribute to these changes, has not been well characterized. The current study describes the loading environment of the lower extremity (LE) during typical days on the International Space Station (ISS) compared to similar data for the same individuals living on Earth. Data from in-shoe force measurements are also used as input to the enhanced daily load stimulus (EDLS) model to determine the mechanical “dose” experienced by the musculoskeletal system and to associate this dose with changes in BMD.Four male astronauts on approximately 6-month missions to the ISS participated in this study. In-shoe forces were recorded using capacitance-based insoles during entire typical working days both on Earth and on-orbit. BMD estimates from the hip and spine regions were obtained from dual energy X-ray absorptiometry (DXA) pre- and post-flight.Measurable loading was recorded for only 30% of the time assigned for exercise. In-shoe forces during treadmill walking and running on the ISS were reduced by 25% and 46%, respectively, compared to similar activities on Earth. Mean on-orbit LE loads varied from 0.20 to 1.3 body weight (BW) during resistance exercise and were ～0.10 BW during bicycle ergometry. Application of the EDLS model showed a mean decrease of 25% in the daily load experienced by the LE. BMD decreased by 0.71% and 0.83% per month during their missions in the femoral neck and lumbar spine, respectively.Our findings support the conclusion that the measured ISS exercise durations and/or loading were insufficient to provide the loading stimulus required to prevent bone loss. Future trials with EDLS values closer to 100% of Earth values will offer a true test of exercise as a countermeasure to on-orbit bone loss.     

Actions of Neurotoxic β-Amyloid on Calcium Homeostasis and Viability of PC12 Cells Are Blocked by Antioxidants but Not by Calcium Channel Antagonists

Abstract: The fragment of β-amyloid comprised of amino acids 25–35 induces a rapid, concentration-dependent increase in cytosolic free calcium levels in suspensions of PC12 neuronal cells. This action of β-amyloid 25–35 is not altered by pretreatment with the calcium channel blockers nifedipine or cobalt, with the depleter of intracellular calcium stores cyclopiazonic acid, or with the phospholipase C inhibitor neomycin. However, the effects of β-amyloid 25–35 on cytosolic free calcium are absent in calcium-free buffer and are blocked by the antioxidant lazaroid U-83836E and by vitamin E. β-Amyloid 25–35 is also neurotoxic and produces a concentration-dependent reduction in the viability of PC12 cells in culture. The neurotoxic action of β-amyloid is blocked by U-83836E and vitamin E but not by nifedipine or cobalt. These data indicate that both the disruption of calcium homeostasis and the reduction of cell viability produced by β-amyloid in PC12 cells are mediated by free radical-based processes.     

RNA from rat hepatoma cells can activate phenylalanine hydroxylase gene of mouse erythroleukemia cells

Mouse erythroleukemia (MEL) cells do not synthesize any detectable level of phenylalanine hydroxylase and thus do not grow in Tyr- medium. Rat hepatoma cells that constitutively express phenylalanine hydroxylase were treated prior to fusion with MEL cells with biochemical inhibitors to inactivate different macromolecular components of the cells, and the fusion products were selected in Tyr- medium. Continuously growing populations of cells resembling the parental MEL cells and expressing mouse phenylalanine hydroxylase were obtained only when rat hepatoma cells treated with mitomycin or iodoacetamide, which inactivate DNA and SH proteins, respectively, were fused with MEL cells. Fusion of MEL cells with UV-treated rat hepatoma cells did not result in the activation of the mouse phenylalanine hydroxylase gene. UV treatment damages both DNA and RNA. These data suggested that RNA was involved in the regulation of phenylalanine hydroxylase gene. Additional evidence for the role of RNA in the phenylalanine hydroxylase gene regulation was obtained from RNA transfection studies. RNA only from cells which express phenylalanine hydroxylase, such as rat hepatoma cells and MEL cybrids, when introduced into MEL cells by the CaPO4 coprecipitation method, resulted in the permanent activation of the mouse phenylalanine hydroxylase gene.     

Influence of internucleotide phosphate linkage on relative base stacking in 3'-5' and 2'-5' RNA: a circular dichroic spectroscopic study of RNA hexamer AACCUU.

The variations in base stacking interactions of two isomeric RNA hexamers, 3'-5'r (AACCUU) and 2'-5'r' (AACCUU), have been studied using temperature dependent CD spectroscopy. Both RNA hexamers, in single strand form, exhibited a right handed helical sense. Van't Hoff analysis of the CD spectral results, derived from a two state model, gave a higher enthalpy of stacking for 3'-5' RNA than for 2'-5'RNA. The results suggest that 3'-5' linkage in RNA facilitates formation of better helical stacks in relation to an isomeric 2'-5' linkage.     

Intercalation of aflatoxin B1 in two oligodeoxynucleotide adducts: comparative 1H NMR analysis of d(ATCAFBGAT).d(ATCGAT) and d(ATAFBGCAT)2

8,9-Dihydro-8-(N7-guanyl-[d(ATCGAT)])-9-hydroxyaflatoxin B1.d(ATCGAT) and 8,9-dihydro-8-(N7-guanyl-[d(ATGCAT)])-9-hydroxyaflatoxin B1.8,9-dihydro-8-(N7-guanyl-[d(ATGCAT)])-9-hydroxyaflatoxin B1 were prepared by direct addition of afltoxin B1 8,9-epoxide to d(ATCGAT)2 and d(ATGCAT)2, respectively. In contrast to reaction of aflatoxin B1 8,9-epoxide with d(ATCGAT)2 which exhibits a limiting stoichiometry of 1:1 aflatoxin B1:d(ATCGAT)2 [Gopalakrishnan, S., Stone, M. P., & Harris, T. M. (1989) J. Am. Chem. Soc. 111, 7232-7239], reaction of aflatoxin B1 8,9-epoxide with d(ATGCAT)2 exhibits a limiting stoichiometry of 2:1 aflatoxin B1:d(ATGCAT)2. 1H NOE experiments, nonselective 1H T1 relaxation measurements, and 1H chemical shift perturbations demonstrate that in both modified oligodeoxynucleotides the aflatoxin moiety is intercalated above the 5'-face of the modified guanine. The oligodeoxynucleotides remain right-handed, and perturbation of the B-DNA structure is localized adjacent to the adducted guanine. Aflatoxin-oligodeoxynucleotide 1H NOEs are observed between aflatoxin and the 5'-neighbor base pair and include both the major groove and the minor groove. The aflatoxin methoxy and cyclopentenone ring protons face into the minor groove; the furofuran ring protons face into the major groove. No NOE is observed between the imino proton of the modified base pair and the imino proton of the 5'-neighbor base pair; sequential NOEs between nucleotide base and deoxyribose protons are interrupted in both oligodeoxynucleotide strands on the 5'-side of the modified guanine. The protons at C8 and C9 of the aflatoxin terminal furan ring exhibit slower spin-lattice relaxation as compared to other oligodeoxynucleotide protons, which supports the conclusion that they face into the major groove. Increased shielding is observed for aflatoxin protons; chemical shift perturbations of the oligodeoxynucleotide protons are confined to the immediate vicinity of the adducted base pair. The imidazole proton of the modified guanine exchanges with water and is observed at 9.75 ppm. The difference in reaction stoichiometry is consistent with an intercalated transition-state complex between aflatoxin B1 8,9-epoxide and B-DNA. Insertion of aflatoxin B1-8,9 epoxide above the 5'-face of guanine in d(ATCGAT)2 would prevent the binding of a second molecule of aflatoxin B1 8,9-epoxide. In contrast, two intercalation sites would be available with d(ATGCAT)2.(ABSTRACT TRUNCATED AT 400 WORDS)