A unique way of energy conservation in glutamate fermenting clostridia

Genetic analysis revealed that Rhodobacter capsulatus contains six rnfABCDEG-genes that are responsible for the electron flow in nitrogen fixation (rnf = Rhodobacter nitrogen fixation). Homolgous genes have been detected in Clostridium tetani. In this work, a membrane complex has been purified from the related Clostridium tetanomorphum that catalyses the reduction of NAD + (E°' = −320 mV) with ferredoxin (E°' ≤ −420 mV). The difference in the redox potential of ≥ 100 mV could be useful for additional energy conservation in the fermentation of glutamate to ammonia, CO 2 , acetate, butyrate, and H 2 . The complex consists of six subunits (RnfABCDEG), of which four N-termini (RnfCDEG) could be sequenced. The sequences are 60-80% identical to the deduced sequences of the Rnf-subunits from C. tetani. The rnf operon has been completely sequenced and aligned with the sequences of C. tetani. The complex contains both non-covalently bound flavin as well as covalently bound flavin. The non-covalently bound flavin was identified as FMN and riboflavin in 1:1 stochiometric ratio, each 0.3 mol/mol Rnf complex (180 kDa). The subunits RnfG and RnfD contain covalently bound flavin linked via phosphodiester bond. The iron was determined as 25±1 mol per Rnf complex. Usually, Rnf activity was measured with NADH and ferricyanide at 420 nm. In order to measure NAD + reduction with reduced ferredoxin catalysed by Rnf complex, the ferredoxin was purified from C. tetanomorphum and reduced by Ti(III)citrate at pH 7.0. High Rnf activities were observed in the membrane preparations of Clostridium aminobutyricum, Clostridium pascui and Clostridium propionicum. Thus, additional energy conservation can be explained in these bacteria. However Rnf activity was absent in Eubacterium barkeri, a nicotinate fermenting bacteria. The soluble butyryl-CoA-dehydrogenase/electron transferring flavoprotein (Bcd/Etf) complex was purified from C. pascui as well as from C. tetanomorphum. The N- terminal sequences of the three subunits (αβγ) showed high identities with the deduced sequences of C. tetani. The Bcd/Etf complex purified from C. tetanomorphum was shown to catalyze the endergonic reduction of ferredoxin with NADH coupled to the exergonic reduction of crotonyl-CoA to butyryl-CoA (E°' = -10 mV) with NADH. The12 reduced ferredoxin could be used for H 2 production catalysed by a hydrogenase or probably used for additional energy conservation via Rnf (about 0.3 mol ATP/ mol glutamate). Experiments with [2,4,4- 2 H] glutamate and detection of citramalate-lyase activity showed that C. pascui and C. tetanomorphum ferment glutamate via the methylaspartate pathway

Whole Animal Automated Platform for Drug Discovery against Multi-Drug Resistant Staphylococcus aureus

Staphylococcus aureus, the leading cause of hospital-acquired infections in the United States, is also pathogenic to the model nematode Caenorhabditis elegans. The C. elegans-S. aureus infection model was previously carried out on solid agar plates where the bacteriovorous C. elegans feeds on a lawn of S. aureus. However, agar-based assays are not amenable to large scale screens for antibacterial compounds. We have developed a high throughput liquid screening assay that uses robotic instrumentation to dispense a precise amount of methicillin resistant S. aureus (MRSA) and worms in 384-well assay plates, followed by automated microscopy and image analysis. In validation of the liquid assay, an MRSA cell wall defective mutant, MW2ΔtarO, which is attenuated for killing in the agar-based assay, was found to be less virulent in the liquid assay. This robust assay with a Z’-factor consistently greater than 0.5 was utilized to screen the Biomol 4 compound library consisting of 640 small molecules with well characterized bioactivities. As proof of principle, 27 of the 30 clinically used antibiotics present in the library conferred increased C. elegans survival and were identified as hits in the screen. Surprisingly, the antihelminthic drug closantel was also identified as a hit in the screen. In further studies, we confirmed the anti-staphylococcal activity of closantel against vancomycin-resistant S. aureus isolates and other Gram-positive bacteria. The liquid C. elegans – S. aureus assay described here allows screening for anti-staphylococcal compounds that are not toxic to the host

Intergenerational relationship and Multi-trait Genotype-Ideotype Distance Indexing (MGIDI) in inter-varietal cross derivatives of black gram [Vigna mungo (L.) Hepper.

Black gram is a highly self-pollinated short-duration crop cultivated in various cropping systems viz., rainfed, intercrop and rice fallow. In the present study, hybridization was made between 2 black gram varieties viz., CO 6 and VBN 11, aimed to develop the recombinants and select superior progenies through Multi-trait Genotype-Ideotype Distance Index (MGIDI) at Tamil Nadu Agricultural University, Coimbatore. The parent-offspring regression analysis was carried out in the F3 and F4 progenies of CO 6 × VBN 11. The significant regression and intergenerational correlation coefficients were observed for all the traits studied. The narrow sense heritability was found to be high for the number of seeds per pod. The other traits viz., plant height, number of branches per plant, number of clusters per plant, number of pods per cluster, number of pods per plant, pod length, number of seeds per pod, hundred seed weight and seed yield per plant showed moderate narrow sense heritability. The MGIDI was calculated for 62 F4 progenies of CO 6 × VBN 11 considering nine yield component traits. In F4 progenies, high broad sense heritability was observed for all the yield-contributing traits and high selection gain was recorded for seed yield per plant (49.77%), number of branches per plant (46.29%), number of pods per plant (37.74%) and number of pods per cluster (20.80%). Based on the MGIDI, six F4 progenies viz., P1, P6, P22, P49, P50 and P53 were selected. The selected progenies could be further evaluated and yield tested for commercial exploitation

Genetic diversity as assessed by morphological and microsatellite markers in greengram (Vigna radiata L.)

Genetic diversity in 20 elite greengram [Vigna radiata (L.) R. Wilczek] genotypes were studied using morphological and microsatellite markers. 18 morphological traits generated 50 alleles and the number of alleles ranged from two to five. 16 microsatellite markers from greengram, adzuki bean, common bean and cowpea were successfully amplified across 20 greengram genotypes of which 14 showed polymorphism. A total of 41 alleles were generated with an average of 2.56 alleles per locus. The number of alleles ranged from two to five. Similarity measures and clustering analyses were made using 91 alleles generated from morphological and microsatellite data. The resulting dendrogram distributed 20 greengram genotypes into six main clusters. The results of the principal component analysis were well congruent with the dendrogram. In the dendrogram as well as in the principal component analyses, genotype Vigna umbellata was placed separately from the rest of the genotypes. There was successful cross species amplification of adzuki bean, common bean and cowpea microsatellite or simple sequence repeat (SSR) markers in greengram. This study demonstrates that a combination of morphological and molecular markers increases the efficiency of diversity measured and the adzuki bean microsatellite markers are highly polymorphic and can be successfully used for genome analysis in greengram. Results indicate that sufficient variability is present in the greengram genotypes studied and would be helpful in the selection of suitable parents for breeding purposes and gene mapping studies.Key words: Greengram, Vigna radiata, morphological markers, cross species amplification, microsatellite markers, genetic diversity

Exploring intra-allelic and inter-allelic gene interactions influencing seed yield and its components in inter-varietal crosses of Mungbean (Vigna radiata (L.) Wilczek)

Mungbean (Vigna radiata (L.) Wilczek) is a versatile legume widely cultivated for its nutritional value and adaptability. Meeting the increasing global demand for nutritious food requires the development of high-yielding varieties. Therefore, understanding the inheritance of yield and component traits is crucial for defining effective breeding strategies. In the present study, we aimed to investigate the genetic effects and interactions governing inheritance through generation mean analysis. The four crosses viz., IPM409-4×VGG18-002, IPM409-4×WGG42, COGG13-39×VGG16-058 and COGG13-39×VGG18-002 and their five generations (P1, P2, F1, F2, and F3) were evaluated for nine yield and yield component traits during 2023 summer season. The significance of additive, dominance, and epistatic components viz.,additive×additive [i] and dominance×dominance [l] of each trait was found to be different among all the crosses. Mungbean is a self-pollinated crop, so only fixable gene effects can be exploited for trait improvement. In the IPM409-4×VGG18-002 cross, all the traits exhibited additive or additive × additive gene action except for plant height and seed yield per plant (dominance). The scaling test was significant in IPM409-4×WGG42 cross for all the traits, except for the number of pods per cluster. Except for the number of branches per plant in which the dominance effect was evident, additive or additive×additive gene effects were observed for the other traits.  In COGG13-39×VGG16-058 and COGG13-39×VGG18-002 crosses, all the yield traits recorded fixable (additive and additive×additive) gene effects except for number of pods per plant in COGG13-39×VGG18-002. Considering the results of all four crosses, gene actions that exhibit consistency across crosses revealed that epistatic interaction (additive×additive) significantly influenced the expression of various mung bean traits. Therefore, the later generation selection of short-duration segregants with high yield, bold seeds, and resistance to yellow mosaic disease from the above populations can be carried out to develop commercially valuable mung bean varieties

Comparative metabolic profiling of resistant and susceptible mungbean (Vigna radiata L. Wilczek) genotypes to elucidate the defense response against mungbean yellow mosaic virus (MYMV) disease

Mungbean Yellow Mosaic Virus (MYMV) disease significantly impacts mungbean crop productivity, with the losses ranging from 10 to 100 percent. Developing host plant resistance offers a sustainable solution to mitigate this challenge. The metabolic changes underlying resistance to MYMV remain primarily unexplored in mungbean. The present study used nontargeted metabolomic profiling to analyze the comparative metabolic changes in resistant and susceptible genotypes upon disease incidence. The methanol extract of leaf samples collected from MYMV disease resistant (GAM 5) and susceptible (ADT 3) genotypes upon occurrence of MYMV disease were subjected to gas chromatography – mass spectrophotometry (GC-MS) analysis. Metabolic profiling resulted in the identification of 40 and 49 metabolites in resistant and susceptible genotypes, respectively. The fold change analysis revealed that 12 metabolites showed significant differences in the abundance level between resistant and susceptible genotypes. Out of 12, nine metabolites were significantly up-regulated in the resistant genotype compared to the susceptible genotype. For all the up-regulated metabolites except Erythrodiol, their role in plant-pathogen interaction was identified as either antimicrobial (ethylene glycol, chlorogenic acid, trifolin), antiviral activity (diphenyl sulfone, 2-amino oxazole), antifeedant (betulin), changes in the specific biochemical and structural property (xylose) or involvement in signaling cascade (oleic acid). These metabolites act as a metabolic biomarker; their interaction with specific molecular targets associated with MYMV infection can be further examined and utilized to rapidly develop MYMV-resistant cultivars in mungbean

Studies on the antagonistic effect of rhizobacteria against soilborne Phytophthora species on strawberry

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Isolation of Mycobacterium Tuberculosis in Body Fluids and Comparison of Conventional Methods with Polymerase Chain Reaction in a Tertiary Care Hospital Chennai.

INTRODUCTION : Tuberculosis has co-existed with humanity since the days before recorded history and evidence of tuberculosis has been found in the skeletal remains of mummies. Hippocrates not only described the disease but also named it ‘phthisis’ which means to mart or waste away. Tuberculosis remains a major global public health problem. Roughly a third of the world's population has been infected with M. tuberculosis, and new infections occur at a rate of one per second. However, not all infections with M. tuberculosis cause TB disease and many infections are asymptomatic. As per WHO 2009 report, in 2008, there were an estimated 8.9-9.9 million incident cases of Tuberculosis, 9.6-13.3 million prevalent cases of Tuberculosis 1.1-1.7 million deaths from Tuberculosis. India had the largest total incidence, with an estimated 2.0 million new cases. India has the most number of cases of tuberculosis and accounts for about one-fifth of the global TB burden and 2/3th of the South –East Asian countries burden. Nearly 40% of the Indian population is affected with tuberculosis bacillus. Each year, 1.9 million cases of TB are detected, amongst which 0.8 million are smear positive new cases (infectious cases). Daily, nearly 1000 people die due to TB (2 deaths/ 3 minutes). As per WHO estimate, there were nearly 322,000 TB deaths in 2006 in India which accounted for 26% of all the preventable adult deaths. AIMS AND OBJECTIVES : Study of prevalence of extra pulmonary tuberculosis in a Tertiary Care Hospital, Chennai Comparison of smear by Ziehl-Neelsen and Auramine Phenol Fluorescence Staining Isolation of Mycobacterium in Extra Pulmonary specimens by employing conventional bacteriological methods. Comparison of culture by Kirchner’s and Lowenstein-Jensen Medium Identification of Mycobacteria by employing standard phenotypic methods. Performing drug susceptibility on the isolates identified as Mycobacterium tuberculosis. Comparison of conventional methods with molecular method IS 6110 based Polymerase Chain Reaction. CONCLUSION A high index of suspicion is necessary to diagnose of Extra Pulmonary Tuberculosis. Delay in diagnosis leads to sequelae in severe forms of Tuberculosis. More than one diagnostic procedure is often needed for confirmation of the diagnosis, as Extra Pulmonary Specimens were paucibacillary in nature. Staining is a rapid method for detection of Mycobacterium Tuberculosis but always it should be confirmed with culture in Lowenstein-Jensen medium which is the Gold standard conventional method. Bacteriological and molecular methods aid in correct diagnosis of Extra Pulmonary Tuberculosis. Drug susceptibility on culture positives will detect Multi Drug Resistant tuberculosis. Mycobacteria grow slowly and replicate every 18-20 hours, so growth in culture requires days or weeks. Culture is laborious and time consuming. The Rapid detection of bacilli with IS6110 based PCR assay is needed in management and prevention of complication

Design and Trajectory Planning of Bipedal Walking Robot with Minimum Sufficient Actuation System

This paper presents a new type of mechanism and trajectory planning strategy for bipedal walking robot. The newly designed mechanism is able to improve the performance of bipedal walking robot in terms of energy efficiency and weight reduction by utilizing minimum number of actuators. The usage of parallelogram mechanism eliminates the needs of having an extra actuator at the knee joint. This mechanism works together with the joint space trajectory planning in order to realize straight legged walking which cannot be achieved by conventional inverse kinematics trajectory planning due to the singularity. The effectiveness of the proposed strategy is confirmed by computer simulation results

Mathematical/Statistical Research of Improved Metal Foam Heat Sink with Fe3O4-H2O Nanofluids

This paper presents a numerical investigation of a proposed heat sink equipped with enhanced metal foam subjected to forced convection. The two-phase Eulerian model is employed to predict the behavior of Fe3O4-H2O nanofluid, to analyze heat transfer properties and entropy production. The simulation results are validated against existing data, and good agreement is achieved. The impact of pore permeability, nanoparticle size, concentration, and flow velocity on heat exchange and entropy is studied. Our results show that the application of reinforced foam enhances average Nusselt by 5.79% compared to aluminum foam, and the proposed foam application can reduce thermal entropy by 47.58% to 81.18% for Re values of 2600 and 3800, respectively. Moreover, PEC increases by 56% when the pore permeability and flow velocity are raised