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Objective The aim of this study was to investigate the normal tissue complication probability (NTCP) for parotid gland of ca tongue patients from two different radiobiological models and to establish a dosimetric validity. Methods Lyman–Kutcher–Burman (LKB) model and linear quadratic (LQ) model were considered for determination of NTCP and tolerance dose (TD5/5 and TD50/5) for the parotid gland of 67 number of ca tongue patients who were treated with the volumetric arc therapy (VMAT) technique. An in‐house developed software on Excel (VBA) was used for this study. Results The equivalent uniform dose (EUD) was linearly proportional to the effective volume (veff) for parotid glands and there was a strong correlation between EUD and veff. At EUD = 46 to 47 Gy, the NTCP of parotid was 0.5 for both the models. The tolerance doses, TD5/5 (veff) and TD50/5 (veff) were exponentially reduced with increase of veff for LKB model; whereas these parameters were volume‐independent in the LQ model. TD5/5 (veff) and TD50/5 (veff) were 31.98 Gy and 45.98 Gy respectively for all 67 patients in the LQ model. Below TD50/5, NTCP of LKB model was less than the NTCP , calculated from LQ model. Conclusion One may consider radiobiological LQ model for estimation of clinical tolerance dose for OARs. Due to lack of clinical data, there are inaccuracy in determination of NTCP from LQ model. If sufficient number of tolerance data for partial volumes are available, the prediction of NTCP would be more confident.

The contribution of selenium and selenoproteins in prostate cancer etiology remains elusive, potentially due to insufficient information regarding the biochemical pathways in which they are involved. There are twenty-five human selenocysteine-containing proteins or selenoproteins as well as a smaller class of selenium-containing proteins that do not include selenocysteine, and their cancer-associated aberrations, both genetic and functional, have evoked special interest, although their contribution to the metabolic reprogramming of prostate cancers remains has not been extensively studied. While benign prostate tissue exhibits a glycolytic phenotype, neoplastic events restore the truncated tricarboxylic acid cycle and enhance oxidative phosphorylation. Two selenium-containing proteins, selenium binding protein 1 and selenoprotein F, affect prostate cancer phenotypes by modulating tumor cell metabolic profiles with significant effects on mitochondrial biology, including oxidative phosphorylation and ATP synthesis. One of the pathways affected by both proteins is the activation of adenosine monophosphate kinase and its downstream signaling with concomitant induction of glycolysis. This review focuses on highlighting the role of these two proteins in modulating the bioenergetic profile of prostate cancer and in maintaining the metabolic plasticity of these cells rendering growth advantage and possible therapeutic resistance.

The use of dietary polyphenols as antimicrobial agents has gained immense popularity in recent years, although few of them-like tannic acid has limited use in this field of research; one of the main reasons is its restricted access through the bacterial membrane. Dissipating the bacterial membrane potential with a sub-lethal dosage of the protonophore, carbonyl cyanide m-chlorophenyl hydrazone, enhanced the tannic acid-cytotoxicity with subsequent inhibition of aerobic respiration in Pseudomonas aeruginosa strains which otherwise exhibited a minimum response to tannic acid. However, ascorbic acid, an antioxidant and bacterial membrane-stabilizing compound, had rescued the cells from both tannic acid- and CCCP-mediated lethality. The results suggested that dispersing the membrane potential with a protonophore can enhance the antibacterial properties of tannic acid.

The aim of this study was to determine the variation of patient setup uncertainties and clinical target volume (CTV) to planning target volume (PTV) margins for different tumor sites based on clinical data with three-dimensional IGRT couch using pre-treatment verification by KV-CBCT. In this study the tumor sites were divided in four categories: (1) brain (50 patients), (2) head and neck (H&N) (226 patients), (3) thorax (64 patients), and (4) pelvis (82 patients). The brain and head and neck patients were immobilized using thermoplastic three and five clamp mask and individual head-rest in the supine position. Using vac-loc had done the immobilization of thorax patients, and thermoplastic mask was used with feet-fix in the supine position for pelvis patients. All brain and H&N patients underwent first 3 days of pretreatment verification by KV-CBCT imaging and weekly one for remaining fractions, whereas for thorax and pelvis cases, patients underwent daily pretreatment verification by KV-CBCT images for the determination of overall distributions of setup error in the directions of anteroposterior, mediolateral, and craniocaudal, and CTV to PTV margins were analyzed with the help of van Herk’s, Stroom’s, and ICRU formulas. The maximum percentage displacement within ± 3 mm for brain, H&N, thorax, and pelvis cases were 96.5, 93.3, 90.9, and 88.3 in the direction of anterioposterior respectively. The maximum CTV to PTV margin, calculated from van Herk’s formula for the brain, H&N, thorax, and pelvis were 4.939 mm, 4.62 mm, 7.16 mm, and 5.763 mm in the craniocaudal axis respectively. The comparison of CTV to PTV margin in three consecutive years on 2016, 2017, and 2018 showed that for brain and head and neck cases, the margin initially had decreased by 1.03 mm and 1.11 mm, and then it had increased by 0.75 mm and 0.37 mm on 2018 with respect to 2017. But for thorax and pelvis cases, margins were gradually decreased on three consecutive years. For adequate target coverage, van Herk-calculated CTV to PTV margin are used in our institution. To reduce the setup uncertainties and CTV to PTV margin frequently, CBCT is essential. In the yearly comparison of CTV to PTV margin, we observed that for brain and H&N cases were not in descending order due to the differently deformation of head rests which were used for molding and treatment separately.

Objective Our study aimed to assess the set-up errors for image-guided radiotherapy at a newly established tertiary radiation center in India and to establish the departmental protocol of clinical target volume–planning target volume (CTV–PTV) margins for different anatomical sites. Materials and Methods This study enrolled the first 200 patients who were treated with curative intent at All India Institute of Medical Sciences, Patna, from February 2019 to September 2019. Number of patients were 53, 26, 53, 11, 6, 47, and 4 for head and neck, brain, breast, thorax, abdomen, pelvis, and craniospinal irradiation (CSI), respectively. The translational vectors for total 1,463 kV cone-beam computed tomography (CBCT) images were collected from the treatment record. Results For the systematic error, the largest value is found for the thoracic subset on the X and Y directions, and for breast patients on Z axis, whereas the smallest values were found for CSI. For random error, the largest value was found for pelvic in the X, Y direction, and for breast subset on Z axis, whereas the smallest values on X and Z axes were found in the brain and for head and neck on the Y axis. Largest value for systemic error is smaller than 5 mm in all directions and for all anatomical subsets. The highest random error value is 5.07 mm in Y axis for pelvic subset. The largest values for CTV–PTV margin are found for thoracic subset and the smallest for CSI followed by the brain. Significant reduction of set-up error observed for the last hundred patients as compared to the first half of the patient population. Conclusion Use of thermoplastic cast along with breast board and respiratory motion management should be recommended to reduce set-up error for breast and thoracic subset. Six degrees of freedom robotic couch system can also further rectify the set-up error in image-guided radiotherapy.

Defective nitrate signaling in plants causes disorder in nitrogen metabolism, and it negatively affects nitrate transport systems, which toggle between high- and low-affinity modes in variable soil nitrate conditions. Recent discovery of a plasma membrane nitrate transceptor protein NRT1.1—a transporter cum sensor—provides a clue on this toggling mechanism. However, the general mechanistic description still remains poorly understood. Here, we illustrate adaptive responses and regulation of NRT1.1-mediated nitrate signaling in a wide range of extracellular nitrate concentrations. The results show that the homodimeric structure of NRT1.1 and its dimeric switch play an important role in eliciting specific cytosolic calcium waves sensed by the calcineurin-B-like calcium sensor CBL9, which activates the kinase CIPK23, in low nitrate concentration that is, however, impeded in high nitrate concentration. Nitrate binding at the high-affinity unit initiates NRT1.1 dimer decoupling and priming of the Thr101 site for phosphorylation by CIPK23. This phosphorylation stabilizes the NRT1.1 monomeric state, acting as a high-affinity nitrate transceptor. However, nitrate binding in both monomers, retaining the unmodified NRT1.1 state through dimerization, attenuates CIPK23 activity and thereby maintains the low-affinity mode of nitrate signaling and transport. This phosphorylation-led modulation of NRT1.1 activity shows bistable behavior controlled by an incoherent feedforward loop, which integrates nitrate-induced positive and negative regulatory effects on CIPK23. These results, therefore, advance our molecular understanding of adaptation in fluctuating nutrient availability and are a way forward for improving plant nitrogen use efficiency.

Primarily synthesized for chelating metal ions from the surrounding media, the pyomelanin plays an important role in bacterial virulence where it is needed for infection and biofilm formation as well as protection from host immune response. In this study, two out of three phenolic acids, gallic acid, and propyl gallate induced pyomelanin in two clinical isolates of Pseudomonas aeruginosa and inhibited biofilm formation. Ascorbic acid treatment reversed the gallic acid and propyl gallate mediated pyomelanin synthesis without reversing the inhibition of the biofilm formation. mRNA expression study revealed the upregulation of homogentisic acid oxidase enzyme by ascorbic acid treatment, possibly contributing towards the inhibition of pyomelanin synthesis. Tannic acid did not show any antibacterial or pyomelanin-induction activities. The synergistic effect of gallates and ascorbic acid in the inhibition of biofilm formation and associated pyomelanin synthesis was evidenced which needs further studies to establish their antibacterial efficacies, especially against the clinical isolates of Pseudomonas sp.

A small number of extragalactic radio sources disclose a pair of low-surface-brightness radio lobes, known as “wings,” aligned at a certain angle to the primary jets. Such exotic sources are known as “winged” radio sources. Here we report the new identification of a total of 26 “winged” radio sources from the LOFAR Two-meter Sky Survey First Data Release (LoTSS DR1). Out of the 26 “winged” sources, 14 are identified as X-shaped radio galaxies and the remaining 12 as Z-shaped radio galaxies. The available optical counterpart of each radio galaxy is cataloged along with its estimated redshift. Among the 26 sources, 15 candidates are classified as FR-II radio galaxies, and two are classified as FR-I type. For nine candidates, no conclusions are drawn due to their complex morphology. We also calculate the physical parameters such as spectral index, radio luminosity, and power of the sources. We have made a statistical study of the spectral index by combining our estimated value with the spectral index collected from previous works. A mean value of spectral index of 0.71 is obtained.

Polyethylene terephthalate (PET) is extensively used in plastic products, and its accumulation in the environment has become a global concern. Being a non-degradable pollutant, a tremendous quantity of PET-bearing plastic materials have already accumulated in the environment, posing severe challenges towards the existence of various endangered species and consequently threatening the ecosystem and biodiversity. While conventional recycling and remediation methodologies so far have been ineffective in formulating a "green" degradation protocol, the bioremediation strategies-though nascent-are exhibiting greater promises towards achieving the target. Very recently, a novel bacterial strain called Ideonella sakaiensis 201-F6 has been discovered that produces a couple of unique enzymes, polyethylene terephthalate hydrolase and mono(2-hydroxyethyl) terephthalic acid hydrolase, enabling the bacteria to utilize PET as their sole carbon source. With a detailed understanding of the protein structure of these enzymes, possibilities for their optimization as PET degrading agents have started to emerge. In both proteins, several amino acids have been identified that are not only instrumental for catalysis but also provide avenues for the applications of genetic engineering strategies to improve the catalytic efficiencies of the enzymes. In this review, we focused on such unique structural features of these two enzymes and discussed their potential as molecular tools that can essentially become instrumental towards the development of sustainable bioremediation strategies. Degradation PET by wild type and genetically engineered PETase and MHETase. Effect of the MHETase-PETase chimeric protein and PETase expressed on the surface of yeast cells on PET degradation is also shown.

Glutamate dehydrogenase (GDH) is a key enzyme interlinking carbon and nitrogen metabolism. Recent discoveries of the GDH specific role in breast cancer, hyperinsulinism/hyperammonemia (HI/HA) syndrome, and neurodegenerative diseases have reinvigorated interest on GDH regulation, which remains poorly understood despite extensive and long standing studies. Notwithstanding the growing evidence of the complexity of allosteric network behind GDH regulation, identifications of allosteric factors and associated mechanisms are paramount to deepen our understanding of the complex dynamics that regulate GDH enzymatic activity. Combining structural analyses of cryo-electron microscopy data with molecular dynamic simulations, here we show that the cofactor NADH is a key player in the GDH regulation process. Our structural analysis indicates that, binding to the regulatory sites in proximity of the antenna region, NADH acts as a positive allosteric modulator by enhancing both the affinity of the inhibitor GTP binding and inhibition of GDH catalytic activity. We further show that the binding of GTP to the NADH-bound GDH activates a triangular allosteric network, interlinking the inhibitor with regulatory and catalytic sites. This allostery produces a local conformational rearrangement that triggers an anticlockwise rotational motion of interlinked alpha-helices with specific tilted helical extension. This structural transition is a fundamental switch in the GDH enzymatic activity. It introduces a torsional stress, and the associated rotational shift in the Rossmann fold closes the catalytic cleft with consequent inhibition of the deamination process. In silico mutagenesis examinations further underpin the molecular basis of HI/HA dominant mutations and consequent over-activity of GDH through alteration of this allosteric communication network. These results shed new light on GDH regulation and may lay new foundation in the design of allosteric agents.

The head–tail (HT) morphology of radio galaxies is seen for a class of radio sources where the primary lobes are being bent in the intercluster weather due to strong interactions between the radio jets and their respective intracluster medium. A systematic search has been carried out for new HT radio galaxies from the Very Large Array Faint Images of the Radio Sky at Twenty-Centimeters survey database at 1400 MHz. Here, we present a catalog of 717 new HT sources, among which 287 are narrow-angle tail (NAT) sources whose opening angle between the two lobes is less than 90°, and 430 are wide-angle tail (WAT) whose the opening angle between the two lobes is greater than 90°. NAT radio sources are characterized by tails bent in a narrow “V”-like shape; the jet bending in the case of WAT radio galaxies are such that the WATs exhibit wide “C”-like morphologies. Optical counterparts are found for 359 HT sources. We report HT sources with luminosity ranges 1038 ≤ L 1.4 GHz ≤ 1045 erg s−1 and redshifts up to 2.01. The various physical properties of these HT sources are mentioned here. Some statistical studies have been done for this large number of newly discovered HT sources.

Accurate spatio-temporal information about rice growth is an important factor for agronomic management and regional grain yield estimation. In this letter, a unified framework for monitoring and mapping of rice using dense time-series of Sentinel-1 synthetic aperture radar (SAR) images is proposed. A processing chain for such dense time-series Sentinel-1 images is developed with the Google Earth Engine’s cloud computing platform. A dense time-series analysis of backscatter response of rice with different management practices is analyzed. Subsequently, the early and late transplanted rice is classified using a clustering algorithm within this platform. The proposed approach is used to monitor different cultivars of rice in three districts in the state of West Bengal, which is one of the major rice growing regions in India. The classification accuracy is assessed across 150 validation points spanning multiple blocks for the 2017 monsoon season. The Sentinel-1 SAR images acquired up to the early vegetative stage for rice have provided satisfactory classification accuracy with an overall accuracy >85% with $\kappa \sim 0.86$ across different management practices throughout the region.

Specific genetic variations in the gene for the selenium-containing antioxidant protein glutathione peroxidase 1 (GPX1) are associated with the risk of a variety of common diseases, including cancer, diabetes, and cardiovascular disorders. Two common variations have been focused upon, one resulting in leucine or proline at codon 198 and another resulting in 5, 6, or 7 alanine repeats were previously shown to affect the distribution of GPX1 between the cytoplasm and mitochondria. Human MCF7 cells engineered to exclusively express GPX1 with five alanine repeats at amino terminus and proline at codon 198 (A5P) and seven alanine repeats at amino terminus and leucine at codon 198 (A7L), as well as derivatives targeted to the mitochondria by the addition of a mitochondrial localization sequence (mA5P and mA7L) were used to assess the consequences of the expression of these proteins on the cellular redox state and bioenergetics. Ectopic expression of A5P and A7L reduced the levels of reactive oxygen species, and the mitochondrially targeted derivatives exhibited better activity in these assays. Bioenergetics and mitochondrial integrity were assessed by measuring mitochondrial membrane potential, oxygen consumption, adenosine triphosphate (ATP) levels, and the levels of lactate dehydrogenase. The results of these assays indicated distinctively, and sometimes opposing, patterns with regard to differences between the consequences of the expression of A5P, A7L, mA5P, and mA7L. These data provide new information on the consequences of differences in the primary structure and cellular location of GPX1 proteins and contribute to the understanding of how these effects might contribute to human disease.

Two novel pyrene-based Schiff base compounds 4-[(5-pyren-1-yl-thiophen-2-ylmethylene)-amino]-phenol (KSB-1) and 4-[(4-pyren-1-yl-benzylidene)-amino]-phenol (KSB-2) were designed and synthesized by a condensation reaction with a diminutive modification of π-spacers. The KSB-1 and KSB-2 molecules are thoroughly characterized by pivotal spectroscopic techniques. The prepared products were found to yield excellent quantity, and crystallographic data was also analyzed for KSB-1. Moreover, the antibacterial activities of the synthesized KSB-1 and KSB-2 were investigated against the two strains of Pseudomonas aeruginosa. BSA binding interaction with these pyrene-based Schiff bases was investigated by fluorescence quenching method, and the quenching of BSA followed a static type. The binding constant and number of binding modes were calculated.

X-shaped radio galaxies are a subclass of radio sources that exhibit a pair of secondary low surface brightness radio lobes oriented at an angle to the primary high surface brightness lobes. Sometimes, the secondary low brightened lobes emerge from the edges of the primary high brightened lobes and form a Z-symmetric morphology. We present a systematical search result for X-shaped radio galaxies (XRGs) and Z-shaped radio galaxies (ZRGs) from the VLA Faint Images of the Radio Sky at Twenty-Centimeters (VLA FIRST) Survey at 1.4 GHz. Our search yields a total of 296 number of radio sources, out of which 161 are XRGs and 135 are ZRGs. We have also made optical identification of these sources from the different available literature. J1124+4325 and J1319+0502 are the farthest known XRG and ZRG, respectively. We have estimated spectral index and radio luminosity of these radio sources and made a comparative study with previously detected XRGs and ZRGs. The average value of luminosities for XRGs is higher than that of ZRGs. With the help of a large sample size of the newly discovered XRGs and ZRGs, various statistical properties of these sources are studied. Out of 161 XRGs presented in the current paper, 70% (113) are FR II radio galaxies and 13% (20) are FR I radio galaxies. For 28 XRGs, the morphology is complex and could not be classified. For XRGs, the statistical studies are done on the angle between the major axis and minor axis and the relative size of the major and minor axes. For the ZRGs a statistical study is done on the angular size., Comment: Published in ApJSS

Using the 144 MHz Low-Frequency Array (LOFAR) telescope, We undelook a search for Miscellaneous Radio Galaxies (MRGs) whose morphological structures are different from known structures. Reseaons of study of these type sources due to their unique and peculiar morphology. Here, we report four MRGs from LOFAR Two Metre Sky Survey first data release (LoTSS DR1) at 144 MHz frequency. The MRS is identified manual visual search (MVS) from LoTSS DR1 data catalogue. We also check their radio morphology from other surveys like The NRAO VLA Sky Survey (NVSS) at 1400 MHz. The corresponding optical counterparts are also identified from Sloan Digital Sky Survey (SDSS), where available. We have estimated different physical parameters like spectral index, radio luminosity, radio power of these MRGs.

Miscellaneous Radio Galaxies (MRGs) are very rare kind of radio galaxies that exhibit unusual and different jet alignment and orientation from a typical radio galaxy. The peculiar and unique morphology of this type of radio source makes them a special case of study. In this paper, we report the identification of fifteen MRGs from VLA Faint Images of the Radio Sky at Twenty-Centimeters (VLA FIRST) Survey. The MRGs are identified manually by visual inspection of the FIRST database (December, 2017). The individual radio morphology of each sources are cross-checked at different frequencies from other surveys like TIFR GMRT Sky Survey at 150 MHz and Westerbork Northern Sky Survey at 325 MHz. We also identify the associated optical counterparts from the Sloan Digital Sky Survey (SDSS), where available. The different physical parameters like spectral index and radio luminosity of these MRGs are also estimated. An overview on the origin of these unique sources are also drawn.

Aim: To understand the effects of gallic acid (GA) on ampicillin (Amp) sensitive or resistant strain of Pseudomonas sp. and also in modulating the corresponding biofilms. Methodology: The cell viability was determined by broth dilution, dry weight and CFU assays. Biofilm formation was measured by crystal violet assay while oxygen consumption rate was measured to verify the metabolic status of the cells. The membrane damage and drug efflux/accumulation were studied by fluorimetric assays. Results: GA transformed the Amp resistant cells, both planktonic and biofilms, into highly sensitive one by inducing membrane damage and enhancing accumulation of drug, whereas the Amp sensitive cells gained resistance against Amp. Conclusion: Use of GA as an antimicrobial compound should be analyzed more critically depending on the drug dosages, drug sensitivity as well as types of bacterial strains being studied.

Summary: Plant adaptation in variable soil nitrate concentrations involves sophisticated signaling and transport systems that modulate a variety of physiological and developmental responses. However, we know very little about their molecular mechanisms. It has recently been reported that many of these responses are regulated by a transceptor NRT1.1, a transporter cum receptor of nitrate signaling. NRT1.1 displays dual-affinity modes of nitrate binding and establishes phosphorylated/non-phosphorylated states at the amino acid residue threonine 101 in response to fluctuating nitrate concentrations. Here we report that intrinsic structural asymmetries between the protomers of the homodimer NRT1.1 provide a functional basis for having dual-affinity modes of nitrate binding and play a pivotal role for the phosphorylation switch. Nitrate-triggered local conformational changes facilitate allosteric communications between the nitrate binding and the phosphorylation site in one protomer, but such communications are impeded in the other. Structural analysis therefore suggests the functional relevance of NRT1.1 interprotomer asymmetries. : Computational Molecular Modeling; Data Analysis in Structural Biology; Plant Biology Subject Areas: Computational Molecular Modeling, Data Analysis in Structural Biology, Plant Biology

In recent years, quantum dots (QDs) are one of the most promising nanomaterials in life sciences community due to their unexploited potential in biomedical applications; particularly in bio-labeling and sensing. In the advanced nanomaterials, carbon dots (CDs) have shown promise in next generation bioimaging and drug delivery studies. Therefore the knowledge of the exact nature of interaction with biomolecules is of great interest to designing better biosensors. In this study, the interaction between CDs derived from tamarind and calf thymus DNA (ct-DNA) has been studied by vital spectroscopic techniques, which revealed that the CDs could interact with DNA via intercalation. The apparent association constant has been deduced from the absorption spectral changes of ct-DNA-CDs using the Benesi-Hildebrand equation. From the DNA induced emission quenching experiments the apparent DNA binding constant of the CDs (Kapp) have also been evaluated. Furthermore, we have analyzed the antibacterial and antifungal activity of CDs using disc diffusion assay method which exhibited excellent activity against E. coli and C. albicans with inhibition zone in the range of 7-12mm. The biocompatible nature of CDs was confirmed by an in vitro cytotoxicity test on L6 normal rat myoblast cells by using MTT assay. The cell viability is not affected till the high dosage of CDs (200μg/mL) for >48h. As a consequence of the work, future development of CDs for microbial control and DNA sensing among the various biomolecules is possible in view of emerging biofields.

Gallic acid and gallate esters are widely used as dietary supplements or additives with clinical significances. Over the last few decades, a large number of publications have been reported stating the antioxidative, antiapoptotic, cardioprotective, neuroprotective, and anticancer properties of gallic acid and gallates, and mostly demonstrated their antioxidative or prooxidative properties influencing the reactive oxygen species (ROS) signaling networks. However, very little focus has been paid to clinical trials, and this restricted their use as a prescribed preventative supplement. Since mitochondria are the principal organelles responsible for ROS generation, we reviewed the existing literature of mitochondria-specific effects of gallates including ROS production, respiration, mitochondrial biogenesis, apoptosis, and the physico-chemical parameters affecting the outcome of gallate supplementation to various health scenarios such as cardiovascular diseases, neurodegeneration, hepatic ailments, or cancers. The major signaling pathways and the molecules targeted by gallic acid and its derivatives have also been discussed with emphasis on mitochondria as the target site. This review provides a better understanding of the effect of gallic acid and gallate esters on mitochondrial functions and in designing effective preventative measures against the onset of various diseases.

Nitrogen is cycled throughout ecosystems by a suite of biogeochemical processes. The high complexity of the nitrogen cycle resides in an intricate interplay between reversible biochemical pathways alternatively and specifically activated in response to diverse environmental cues. Despite aggressive research, how the fundamental nitrogen biochemical processes are assembled and maintained in fluctuating soil redox conditions remains elusive. Here, we address this question using a kinetic modelling approach coupled with dynamical systems theory and microbial genomics. We show that alternative biochemical pathways play a key role in keeping nitrogen conversion and conservation properties invariant in fluctuating environments. Our results indicate that the biochemical network holds inherent adaptive capacity to stabilize ammonium and nitrate availability, and that the bistability in the formation of ammonium is linked to the transient upregulation of the amo-hao mediated nitrification pathway. The bistability is maintained by a pair of complementary subsystems acting as either source or sink type systems in response to soil redox fluctuations. It is further shown how elevated anthropogenic pressure has the potential to break down the stability of the system, altering substantially ammonium and nitrate availability in the soil, with dramatic effects on biodiversity.

Manganese superoxide dismutase (MnSOD) is a mitochondrial-resident enzyme that reduces superoxide to hydrogen peroxide (H2O2), which can be further reduced to water by glutathione peroxidase (GPX1). Data from human studies have indicated that common polymorphisms in both of these proteins are associated with the risk of several cancers, including breast cancer. Moreover, polymorphisms in MnSOD and GPX1 were shown to interact to increase the risk of breast cancer. To gain an understanding of the molecular mechanisms behind these observations, we engineered human MCF-7 breast cancer cells to exclusively express GPX1 and/or MnSOD alleles and investigated the consequences on the expression of several proteins associated with cancer aetiology. Little or no effect was observed on the ectopic expression of these genes on the phosphorylation of Akt, although allele-specific effects and interactions were observed for the impact on the levels of Bcl-2, E-cadherin and Sirt3. The patterns observed were not consistent with the steady-state levels of H2O2 determined in the transfected cells. These results indicate plausible contributing factors to the effects of allelic variations on cancer risk observed in human epidemiological studies.

Background Significant data supports the health benefits of selenium although supplementation trials have yielded mixed results. GPx-1, whose levels are responsive to selenium availability, is implicated in cancer etiology by human genetic data. Selenium's ability to alter the phosphorylation of the H2AX, a histone protein that functions in the reduction of DNA damage by recruiting repair proteins to the damage site, following exposure to ionizing radiation and bleomycin was investigated. Methods Human cell lines that were either exposed to selenium or were transfected with a GPx-1 expression construct were exposed to ionizing radiation or bleomycin. Phosphorylation of histone H2AX was quantified by flow cytometry and survival by the MTT assay. Phosphorylation of the Chk1 and Chk2 checkpoint proteins was quantified by western blotting. Results In colon-derived cells, selenium increases GPx-1 and attenuated H2AX phosphorylation following genotoxic exposures while the viability of these cells was unaffected. MCF-7 cells and transfectants that express high GPx-1 levels were exposed to ionizing radiation and bleomycin, and H2AX phosphorylation and cell viability were assessed. GPx-1 increased H2AX phosphorylation and viability following the induction of DNA damage while enhancing the levels of activated Chk1 and Chk2. Conclusions Exposure of mammalian cells to selenium can alter the DNA damage response and do so by mechanisms that are dependent and independent of its effect on GPx-1. General significance Selenium and GPx-1 may stimulate the repair of genotoxic DNA damage and this may account for some of the benefits attributed to selenium intake and elevated GPx-1 activity.

Impaired glutamate dehydrogenase (GDH) sensitivity to its inhibitors causes excessive insulin secretion by pancreatic beta-cells and defective ammonia metabolism in the liver. These symptoms are commonly associated with hyperinsulinism/hyperammonemia syndrome (HI/HA), which causes recurrent hypoglycaemia in early infancy. Hepatic localization of GDH amination and deamination activities linked with the urea cycle is known to be involved in ammonia metabolism and detoxification. Although deamination activities of hepatic GDH in the periportal zones of liver lobules and its connection to the urea cycle have been exhaustively investigated, physiological roles of GDH amination activity observed at pericentral zones have often been overlooked. Using kinetic modelling approaches, here we report a new role for hepatic GDH amination kinetics in maintaining ammonia homeostasis under an excess intrahepatocyte input of ammonium. We have shown that α-ketoglutarate substrate inhibition kinetics of GDH, which include both random and obligatory ordered association/dissociation reactions, robustly control the ratio between glutamate and ammonium under a wide range of intracellular substrate variation. Dysregulation of this activity under pericentral nitrogen insufficiency contributes to the breaking down of ammonia homeostasis and thereby can significantly affect HI/HA syndrome.

Cancers have long being hallmarked as cells relying heavily on their glycolysis for energy generation in spite of having functional mitochondria. The metabolic status of the cancer cells have been revisited time and again to get better insight into the overall carcinogenesis process which revealed the apparent crosstalks between the cancer cells with the fibroblasts present in the tumour microenvironment. This review focuses on the mechanisms of transformations of normal fibroblasts to cancer-associated fibroblasts (CAF), the participation of the CAF in tumour progression with special interest to the role of CAF cellular glycolysis in the overall tumorigenesis. The fibroblasts, when undergoes the transformation process, distinctly switches to a more glycolytic phenotype in order to provide the metabolic intermediates necessary for carrying out the mitochondrial pathways of ATP generation in cancer cells. This review will also discuss the molecular mechanisms responsible for this metabolic make over promoting glycolysis in CAF cells. A thorough investigation of the pathways and molecules involved will not only help in understanding the process of activation and metabolic reprogramming in CAF cells but also might open up new targets for cancer therapy.

IL (interleukin)-6, an established growth factor for multiple myeloma cells, induces myeloma therapy resistance, but the resistance mechanisms remain unclear. The present study determines the role of IL-6 in re-establishing intracellular redox homoeostasis in the context of myeloma therapy. IL-6 treatment increased myeloma cell resistance to agents that induce oxidative stress, including IR (ionizing radiation) and Dex (dexamethasone). Relative to IR alone, myeloma cells treated with IL-6 plus IR demonstrated reduced annexin/propidium iodide staining, caspase 3 activation, PARP [poly(ADP-ribose) polymerase] cleavage and mitochondrial membrane depolarization with increased clonogenic survival. IL-6 combined with IR or Dex increased early intracellular pro-oxidant levels that were causally related to activation of NF-κB (nuclear factor κB) as determined by the ability of N-acetylcysteine to suppress both pro-oxidant levels and NF-κB activation. In myeloma cells, upon combination with hydrogen peroxide treatment, relative to TNF (tumour necrosis factor)-α, IL-6 induced an early perturbation in reduced glutathione level and increased NF-κB-dependent MnSOD (manganese superoxide dismutase) expression. Furthermore, knockdown of MnSOD suppressed the IL-6-induced myeloma cell resistance to radiation. MitoSOX Red staining showed that IL-6 treatment attenuated late mitochondrial oxidant production in irradiated myeloma cells. The present study provides evidence that increases in MnSOD expression mediate IL-6-induced resistance to Dex and radiation in myeloma cells. The results of the present study indicate that inhibition of antioxidant pathways could enhance myeloma cell responses to radiotherapy and/or chemotherapy.

In the preceding paper (A. Ghosh et al. (2011) Biochemistry (Moscow), 76, 1051-1060), using several comparable tissue materials, it has been convincingly demonstrated that methylglyoxal, a normal metabolite, inhibits mitochondrial complex I of specifically malignant cells. This suggests a distinct alteration of complex I, a highly important enzyme for energy (ATP) production, in malignancy. The present paper shows that as a consequence of this inhibition mitochondrial membrane potential is drastically reduced in sarcoma tissue but not in normal skeletal muscle. This was estimated spectrofluorimetrically using the dye rhodamine 123. As a consequence, cytochrome c was released from the sarcoma mitochondria as evidenced by Western blot analysis. Moreover, on treatment with methylglyoxal membrane potential collapse of sarcoma 180 cells was also indicated by fluorescence-activated cell sorter analysis. Atomic force microscopic study demonstrated gross structural alteration specifically of tumor mitochondria on methylglyoxal treatment. All these studies suggest that methylglyoxal might initiate an apoptotic event in malignant cells.

The creatine/creatine kinase (CK) system plays a key role in cellular energy buffering and transport. In vertebrates, CK has four isoforms expressed in a tissue-specific manner. In the process of creatine biosynthesis several other important metabolites are formed. The anticancer effect of creatine had been reported in the past, and recent literature has reported low creatine content in several types of malignant cells. Furthermore, creatine can protect cardiac mitochondria from the deleterious effects of some anticancer compounds. Previous work from our laboratory showed progressive decrease of phosphocreatine, creatine and CK upon transformation of skeletal muscle into sarcoma. It was convincingly demonstrated that prominent expression of creatine-synthesizing enzymes L-arginine: glycine amidinotransferase and N-guanidinoacetate methyltransferase occurs in sarcoma, Ehrlich ascites carcinoma and sarcoma 180 cells; whereas, both these enzymes are virtually undetectable in skeletal muscle. Creatine transporter also remained unaltered in malignant cells. The anticancer effect of methylglyoxal had been known for a long time. The present work shows that this anticancer effect of methylglyoxal is significantly augmented in presence of creatine. On creatine supplementation the effect of methylglyoxal plus ascorbic acid was further augmented and there was no visible sign of tumor. Moreover, creatine and CK, which were very low in sarcoma tissue, were significantly elevated with the concomitant regression of tumor.