RESUMO
BACKGROUND: The continuous cultivation of rice-wheat in the same field is a key element of double-cropping systems in the Indo-Gangetic plains. Yields of such cropping systems are increasingly challenged as climate change drives increases in temperature, terminal stress and uneven rainfall, delaying rice harvesting and subsequently delaying sowing of wheat. In this paper, we evaluate the optimum sowing dates to achieve high grain yield and quality of wheat cultivars in northwest India. Three cultivars of wheat, HD-2967, HD-3086 and PBW-723, were sown on three different dates at the research farm of ICAR-IARI, New Delhi, to generate different weather conditions at different phenological stages. Different biophysical attributes, photosynthetic rate, stomatal conductance and transpiration rate, were measured at different phenological stages. Yield and grain quality parameters such as protein, starch, amylopectin, amylose and gluten were measured in different cultivars sown on different dates. RESULTS: Biophysical parameters were found to be higher in timely sown crops followed by late-sown and very late-sown crops. Further, the different sowing dates had a significant (P < 0.05) impact on the grain quality parameters such as protein, starch, amylopectin, amylose and gluten content. Percentage increases in the value of starch and amylose content under timely sown were ~7% and 11.6%, ~5% and 8.4%, compared to the very late-sown treatment. In contrast, protein and amylopectin contents were found to increase by ~9.7% and 7.5%, ~13.8% and 16.6% under very late-sown treatment. CONCLUSION: High-temperature stress during the grain-filling periods significantly decreased the grain yield. Reduction in the grain yield was associated with a reduction in starch and amylose content in the grains. The protein content in the grains is less affected by terminal heat stress. Cultivar HD-3086 had higher growth, yield as well as quality parameters, compared to HD-2967 and PBW-723 in all treatments, hence could be adopted by farmers in northwest India. © 2024 Society of Chemical Industry.
Assuntos
Produção Agrícola , Triticum , Triticum/metabolismo , Triticum/crescimento & desenvolvimento , Triticum/química , Triticum/classificação , Índia , Produção Agrícola/métodos , Grão Comestível/química , Grão Comestível/crescimento & desenvolvimento , Grão Comestível/metabolismo , Amido/metabolismo , Amido/análise , Amido/química , Amilose/metabolismo , Amilose/análise , Estações do Ano , Fotossíntese , Amilopectina/metabolismo , Amilopectina/química , Proteínas de Plantas/metabolismo , Sementes/química , Sementes/metabolismo , Sementes/crescimento & desenvolvimento , Agricultura/métodosRESUMO
The radiation arising from naturally occurring radioactive materials (NORMs) is the foremost contributor to the collective dose received by the global population. The present study aims to measure the natural background radiation level and the associated gamma radiation dose in air in the Beldih apatite mine region of Purulia district, India. This study is primarily focused on the determination of 238U, 232Th and 40K activities in the sub-surface soil of the study area. The measurements were carried out using a High Purity Germanium (HPGe) detector-based gamma-ray spectrometer with a relative efficiency of 80%. To achieve uniformity in exposure estimations, radium equivalent activity has been calculated. Additionally, the internal hazard index, external hazard index, radioactivity level index and gamma dose rates have been evaluated to estimate the radiation hazard levels in the study area. The comparison of obtained concentrations and hazard indices with global data (UNSCEAR. (2008). Sources, effects and risks of ionizing radiation. United nations scientific committee on the effects of atomic radiation (report to the general assembly, with Annexes).) suggests that this region lies in a relatively high background radiation zone.
Assuntos
Monitoramento de Radiação , Radioatividade , Rádio (Elemento) , Poluentes Radioativos do Solo , Tório/análise , Radioisótopos de Potássio/análise , Solo , Poluentes Radioativos do Solo/análise , Rádio (Elemento)/análiseRESUMO
Novel CRISPR systems capable of cleaving both DNA and RNA are progressively emerging as attractive tools for genome manipulation of prokaryotic and eukaryotic organisms. We report specific characteristics of CRISPR systems present in Oxynema aestuarii AP17, a halotolerant, filamentous cyanobacterium and the second known member of the Oxynema genus. In-silico analyses of its whole-genome sequence revealed the presence of multiple Type I and Type III CRISPR loci with one Type I-G system previously unreported in cyanobacteria. We further identified the leader sequences at the 5' end of multiple CRISPR loci, many of which were distinct from previously reported cyanobacterial CRISPR leaders. Phylogenetic analyses of the O. aestuarii AP17 Cas1 proteins revealed two protein sequences that were unique and distantly related to other cyanobacterial Cas1 protein sequences. Our findings are significant because novel Class 1 CRISPR systems possess multi-subunit effectors and are highly flexible for repurposing by protein domain fusions made to the effector complex. Additionally, Type III CRISPRs are particularly useful for genome editing in certain extremophiles for which mesophilic Type II CRISPRs are ineffective.
Assuntos
Sistemas CRISPR-Cas , Cianobactérias , Filogenia , Cianobactérias/genética , Genoma , Análise de SequênciaRESUMO
BACKGROUND: Hemifacial spasm (HFS) is a distressing, involuntary, irregular tonic-clonic contraction of the facial muscles innervated by the seventh cranial nerve. It affects the quality of life. Botulinum toxin is a preferred symptomatic treatment option for the condition. However, there is a lack of study in the Indian scenario. Therefore, we observed the demographic profile, clinical spectrum, therapeutic response, and adverse effects of botulinum toxin and assessed the quality of life in the pre and postinjection phases in our subjects with HFS. MATERIALS AND METHODS: The study design is a prospective open-label observational study. Consecutive cases of HFS were selected from the general neurology outpatient department (OPD) and movement disorder clinic of a medical college hospital in Eastern India. Clinical and relevant neuroimaging studies excluded mimickers and secondary causes of HFS. Institutional Ethics Committee's permission was obtained. Informed consent was taken from patients before botulinum toxin injection. The pre and postinjection assessment tools were spasm rate for a specific period of time, quantification of facial asymmetry, widening palpebral fissure by visual analog scale, Jankovic disability rating scale, HFS-7 scale, and videography. RESULTS: A total of 250 cases of HFS (F:M = 138:112) were studied. The mean age of presentation was 47 years. The mean dose of botulinum toxin injection was 24.2 units per patient. The mean duration of improvement was 4 months. The spasm frequency was decreased by 90%, and the facial asymmetry was improved by 86%. The improvement in quality of life was 86%. Local adverse effects are seen in 10.4% of cases, and all were reversible. CONCLUSION: This is one of the largest studies on the effects of botulinum toxin in subjects with HFS in the Indian population. Periodic injection of botulinum toxin is a safe and effective therapy for subjects with HFS. There is a significant improvement in the quality of life following botulinum toxin therapy in subjects with HFS. Adverse effects were local, mild, well-tolerated, and reversible.
Assuntos
Toxinas Botulínicas Tipo A , Espasmo Hemifacial , Fármacos Neuromusculares , Humanos , Pessoa de Meia-Idade , Toxinas Botulínicas Tipo A/efeitos adversos , Espasmo Hemifacial/tratamento farmacológico , Fármacos Neuromusculares/efeitos adversos , Qualidade de Vida , Assimetria Facial , Estudos Prospectivos , ÍndiaRESUMO
The discovery of new antimicrobials is the prime target in the fight against antimicrobial resistance. The continuous search for new lead compounds from bacteria of untapped and extreme ecosystems such as mangroves is currently being undertaken. This study describes the metabolite profiling of the Streptomyces euryhalinus culture extract. Previously, Streptomyces euryhalinus was isolated from the mangrove forest of Indian Sundarbans as a novel microorganism. The antimicrobial mechanism of action of Streptomyces euryhalinus culture extract against bacteria and fungi has been analyzed in this study. The gas chromatography-mass spectrometry profile of the ethyl acetate extract bacterial culture displayed the presence of several bioactive compounds with antibacterial, antifungal and antioxidant properties. The bacterial extract showed significant antimicrobial activity in terms of zone of inhibition, minimum inhibitory concentration, minimum bactericidal concentration, and minimum fungicidal concentration. Moreover, substantial capacity to alter or damage the inner membrane as well as the outer membrane of the gram-positive and gram-negative bacteria was exhibited by the bacterial extract. This membrane alteration or damaging potential of the extract is the mechanism of action. Biofilm formation inhibition property of the extract also signified its antimicrobial action, and possible use against resistant bacteria. The extract has shown notable activity against the virulence factors like prevention of hemolysis in bacteria and inhibition of secreted aspartyl proteinase in fungi. These functions of the bacterial extract have revealed the extent of its action in the prevention of infection by terminating the secretory virulence factors and by damaging the tissue.
Assuntos
Antibacterianos , Ecossistema , Antibacterianos/farmacologia , Bactérias Gram-Negativas , Bactérias Gram-Positivas , StreptomycesRESUMO
An experiment was conducted in the Free Air Ozone and Carbon dioxide Enrichment (FAOCE) facility to study the impact of elevated O3, CO2 and their interaction on chickpea crop (cv. Pusa-5023) in terms of phenology, biophysical parameters, yield components, radiation interception and use efficiency. The crop was exposed to elevated O3 (EO:60ppb), CO2 (EC:550 ppm) and their combined interactive treatment (ECO: EC+EO) during the entire growing season. Results revealed that the crop's total growth period was shortened by 10, 14 and 17 days under elevated CO2, elevated O3 and the combined treatment, respectively. Compared to ambient condition, the leaf area index (LAI) under elevated CO2 was higher by 4 to 28%, whilst it is reduced by 7.3 to 23.8% under elevated O3. The yield based radiation use efficiency (RUEy) was highest under elevated CO2 (0.48 g MJ-1), followed by combined (0.41 g MJ-1), ambient (0.38 g MJ-1) and elevated O3 (0.32 g MJ-1) treatments. Elevated O3 decreased RUEy by 15.78% over ambient, and the interaction results in a 7.8% higher RUEy. The yield was 31.7% more under elevated CO2 and 21.9% lower in elevated O3 treatment as compared to the ambient. The combined interactive treatment recorded a higher yield as compared to ambient by 9.7%. Harvest index (HI) was lowest under elevated O3 (36.10%), followed by ambient (39.18%), combined (40.81%), and highest was under elevated CO2 (44.18%). Chickpea showed a positive response to elevated CO2 resulting a 5% increase in HI as compared to ambient condition. Our findings quantified the positive and negative impacts of elevated O3, CO2 and their interaction on chickpea and revealed that the negative impacts of elevated O3 can be compensated by elevated CO2 in chickpea. This work promotes the understanding of crop behaviour under elevated O3, CO2 and their interaction, which can be used as valuable inputs for radiation-based crop simulation models to simulate climate change impact on chickpea crop.
Assuntos
Cicer , Ozônio , Dióxido de Carbono , Ozônio/farmacologia , Folhas de Planta , Estações do AnoRESUMO
A novel Gram-positive and endospore-forming bacterium assigned as strain SPB7T which is also a new source of a cyclic diketopiperazine (3S,6S)-3,6-diisobutylpiperazine-2,5-dione is described. A polyphasic (biochemical, phenotypic and genotypic) approach was used to clarify the taxonomic affiliation of this strain. The partial and complete 16S rRNA gene sequences revealed that strain SPB7T is a member of the Bacillus genus [showing high similarity (> 98.70%) with Bacillus spizizenii NRRL B-23049T, Bacillus tequilensis KCTC 13622T, Bacillus inaquosorum KCTC 13429T and Bacillus cabrialesii TE3T]. The maximum values for average nucleotide identity (ANI) and in silico DNA-DNA hybridization (GGDC, Formula 2) of strain SPB7T was obtained for twenty-five strains of Bacillus spizizenii (ANI 95.01-95.48% and GGDC 62.70-60.00%). The whole-genome phylogenetic relationship showed that SPB7T formed an individual and separated clade with the Bacillus spizizenii group. Principal cellular fatty acids identified in strain SPB7T were anteiso C15:0, anteiso C17:0, iso C15:0, iso C17:0, C16:0, C10:0 3OH and iso C17:1 Ï10c. Polar lipid profile showed presence of diphosphotidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, two unknown phospholipids and five unknown lipids. Cells were rod shaped, catalase, oxidase-positive and motile. Growth occurred at 20-45 °C (optimal 35 °C), at pH 6.0-10.0 (optimal pH 8) and 0-10% (w/v) NaCl (optimal 2%). The phenotypic, biochemical, and genotypic traits of strain SPB7T strongly supported its taxonomic affiliation as a novel species of the Bacillus genus, for which the name Bacillus rugosus sp. nov. is proposed. The type strain is SPB7T (= NRRL B-65559T, = CICC 24827T, = MCC 4185T).
Assuntos
Anti-Infecciosos/metabolismo , Bacillus/isolamento & purificação , Bacillus/metabolismo , Dicetopiperazinas/metabolismo , Poríferos/microbiologia , Animais , Bacillus/classificação , Bacillus/genética , Técnicas de Tipagem Bacteriana , DNA Bacteriano/genética , Lipídeos/análise , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNARESUMO
Vorinostat is a histone deacetylase (HDAC) inhibitor that inhibits cell proliferation and induces apoptosis in solid tumors, and is in clinical trials for the treatment of glioblastoma (GBM). The goal of this study was to assess whether hyperpolarized 13 C MRS and magnetic resonance spectroscopic imaging (MRSI) can detect HDAC inhibition in GBM models. First, we confirmed HDAC inhibition in U87 GBM cells and evaluated real-time dynamic metabolic changes using a bioreactor system with live vorinostat-treated or control cells. We found a significant 40% decrease in the 13 C MRS-detectable ratio of hyperpolarized [1-13 C]lactate to hyperpolarized [1-13 C]pyruvate, [1-13 C]Lac/Pyr, and a 37% decrease in the pseudo-rate constant, kPL , for hyperpolarized [1-13 C]lactate production, in vorinostat-treated cells compared with controls. To understand the underlying mechanism for this finding, we assessed the expression and activity of lactate dehydrogenase (LDH) (which catalyzes the pyruvate to lactate conversion), its associated cofactor nicotinamide adenine dinucleotide, the expression of monocarboxylate transporters (MCTs) MCT1 and MCT4 (which shuttle pyruvate and lactate in and out of the cell) and intracellular lactate levels. We found that the most likely explanation for our finding that hyperpolarized lactate is reduced in treated cells is a 30% reduction in intracellular lactate levels that occurs as a result of increased expression of both MCT1 and MCT4 in vorinostat-treated cells. In vivo 13 C MRSI studies of orthotopic tumors in mice also showed a significant 52% decrease in hyperpolarized [1-13 C]Lac/Pyr when comparing vorinostat-treated U87 GBM tumors with controls, and, as in the cell studies, this metabolic finding was associated with increased MCT1 and MCT4 expression in HDAC-inhibited tumors. Thus, the 13 C MRSI-detectable decrease in hyperpolarized [1-13 C]lactate production could serve as a biomarker of response to HDAC inhibitors.
Assuntos
Espectroscopia de Ressonância Magnética Nuclear de Carbono-13 , Glioblastoma/diagnóstico por imagem , Glioblastoma/enzimologia , Inibidores de Histona Desacetilases/farmacologia , Imageamento por Ressonância Magnética , Acetilação , Animais , Reatores Biológicos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Feminino , Histonas/metabolismo , Ácido Láctico/biossíntese , Metaboloma/efeitos dos fármacos , Camundongos Nus , Transportadores de Ácidos Monocarboxílicos/metabolismo , Proteínas Musculares/metabolismo , Ácido Pirúvico/metabolismo , Análise de Sobrevida , Simportadores/metabolismo , Vorinostat/farmacologiaRESUMO
Heavy metals such as lead, chromium, and metalloid like arsenic dominate the pinnacle in posing a threat to life. Being environment-friendly, elucidating the mechanism by which microorganisms detoxify such elements has always been an active field of research hitherto. In the present study, we have investigated the capability of nitrogen-deprived Papiliotrema laurentii strain RY1 toward enhanced tolerance and neutralizing toxic elements. There were biosorption and bioprecipitation of lead and chromium at the cell surfaces. Bioprecipitation mechanisms included the formation of lead phosphates and pyromorphites from lead, grimaldite from chromium. Transcripts such as metallothionein, aquaporins, and arsenical pump-driving ATPase have been surmised to be involved in the detoxification of elements. Furthermore, activation of antioxidant defense mechanisms for the cells for each of the elements should contribute towards yeast's propagation. The efficiency of removal of elements for live cells and immobilized cells were high for lead and chromium. To the best of our knowledge, this is the first report of such high tolerance of lead, arsenic, and chromium for any yeast. The yeast showed such varied response under dual stress due to nitrogen starvation and in the presence of respective elements. The yeast possesses promising potentials in nitrogen deprived and enriched environments to aid in bioremediation sectors.
Assuntos
Arsênio/metabolismo , Basidiomycota/metabolismo , Poluentes Ambientais/metabolismo , Metais Pesados/metabolismo , Nitrogênio/metabolismo , Antioxidantes/metabolismo , Arsênio/toxicidade , Basidiomycota/efeitos dos fármacos , Basidiomycota/crescimento & desenvolvimento , Biodegradação Ambiental , Transporte Biológico/genética , Cádmio/metabolismo , Cádmio/toxicidade , Poluentes Ambientais/toxicidade , Expressão Gênica , Inativação Metabólica , Chumbo/metabolismo , Chumbo/toxicidade , Metalotioneína/genética , Metais Pesados/toxicidade , Testes de Sensibilidade MicrobianaRESUMO
The M2 isoform of pyruvate kinase (PK) is upregulated in most cancers including glioblastoma. Although PKM2 has been reported to use dual kinase activities to regulate cell growth, it also interacts with phosphotyrosine (pY)-containing peptides independently of its kinase activity. The potential for PKM2 to use the binding of pY-containing proteins to control tumor growth has not been fully examined. We here describe a novel mechanism by which PKM2 interacts in the nucleus with the RNA binding protein HuR to regulate HuR sub-cellular localization, p27 levels, cell cycle progression and glioma cell growth. Suppression of PKM2 in U87, T98G and LN319 glioma cells resulted in increased p27 levels, defects in entry into mitosis, increased centrosome number, and decreased cell growth. These effects could be reversed by shRNA targeting p27. The increased levels of p27 in PKM2 knock-down cells were caused by a loss of the nuclear interaction between PKM2 and HuR, and a subsequent cytoplasmic re-distribution of HuR, which in turn led to increased cap-independent p27 mRNA translation. Consistent with these results, the alterations in p27 mRNA translation, cell cycle progression and cell growth caused by PKM2 suppression could be reversed in vitro and in vivo by suppression of HuR or p27 levels, or by introduction of forms of PKM2 that could bind pY, regardless of their kinase activity. These results define a novel mechanism by which PKM2 regulates glioma cell growth, and also define a novel set of potential therapeutic targets along the PKM2-HuR-p27 pathway.
Assuntos
Proteínas de Transporte/genética , Proteína Semelhante a ELAV 1/genética , Glioblastoma/genética , Proteínas de Membrana/genética , Antígeno Nuclear de Célula em Proliferação/genética , Hormônios Tireóideos/genética , Proteínas de Transporte/antagonistas & inibidores , Ciclo Celular/genética , Linhagem Celular Tumoral , Proliferação de Células/genética , Proteína Semelhante a ELAV 1/biossíntese , Regulação Neoplásica da Expressão Gênica , Glioblastoma/patologia , Humanos , Proteínas de Membrana/antagonistas & inibidores , Antígeno Nuclear de Célula em Proliferação/biossíntese , RNA Mensageiro/biossíntese , RNA Interferente Pequeno/genética , Proteínas de Ligação a Hormônio da TireoideRESUMO
Phytoremediation, popularly known as 'green technology' has been employed in the present investigation to examine the potential of fluoride removal from water by some aquatic plants. Fluoride contamination in drinking water is very much prevalent in different parts of the world including India. Batch studies were conducted using some aquatic plants e.g., Pistia stratiotes, Eichhornia crassipes, and Spirodela polyrhiza which profusely grow in natural water bodies. The experimental data exhibited that all the above three aquatic floating macrophytes could remove fluoride to some relative degree of efficiency corresponding to initial concentration of fluoride 3, 5, 10, 20 mg/l after 10 days exposure time. Result showed that at lower concentration level i.e., 3 mg/L removal efficiency of Pistia stratiotes (19.87%) and Spirodela polyrhiza (19.23%) was found to be better as compared to Eichhornia crassipes (12.71%). Some of the physiological stress induced parameters such as chlorophyll a, chlorophyll b, total chlorophyll, carotenoid, total protein, catalase, and peroxidase were also studied to explore relative damage within the cell. A marginal stress was imparted among all the plants for lower concentration values (3 mg/L), whereas at 20 mg/l, maximum damage was observed.
Assuntos
Araceae/metabolismo , Eichhornia/química , Recuperação e Remediação Ambiental/métodos , Fluoretos/metabolismo , Poluentes Químicos da Água/metabolismo , Biodegradação Ambiental , Recuperação e Remediação Ambiental/instrumentação , ÍndiaRESUMO
The existing techniques for detection of polyhydroxyalkanoates (PHA) in halophilic archaea/bacteria are either imprecise or require prior PHA production before screening. The proposed method involves amplification of the approximately 280-300 bp conserved region of Class III PHA synthase (phaC) gene of halophiles using the primers codehopCF and codehopCR (Han et al. Appl Environ Microb 76:7811-7819, 2010). In this study, the best reaction condition was ascertained after repeated trials. This developed method was tested on nine haloarchaeal and halobacterial type strains and 28 environmental halophilic archaea and bacteria isolated from the salt pans of the east and west coasts of India. 29 strains were found to be phaC-positive, while eight were found to be phaC-negative although they appeared PHA positive through conventional Nile Red staining. 16S rRNA-based phylogenetic analysis identified 9 haloarchaeal and 9 halobacterial species as novel PHA producers. Multiple sequence alignment of the phaC gene-derived amino acid sequences showed that only 7 amino acid residues were conserved within all four classes of phaC enzymes, whereas 61 amino acids were identical among the phaC enzyme specific to the haloarchaeal and halobacterial strains presently investigated. All phaC-positive strains produced PHA in standard nutrient deficient medium, whereas the phaC-negative strains did not accumulate any PHA as detected by gas chromatography and nuclear magnetic resonance analyses, thus proving the precision of the developed method and elimination of false positives seen with the traditional Nile Red staining procedure.
Assuntos
Aciltransferases/genética , Archaea/genética , Proteínas Arqueais/genética , Proteínas de Bactérias/genética , Euryarchaeota/genética , Aciltransferases/química , Sequência de Aminoácidos , Archaea/enzimologia , Archaea/metabolismo , Proteínas Arqueais/química , Proteínas de Bactérias/química , Euryarchaeota/enzimologia , Euryarchaeota/metabolismo , Microbiologia Industrial/métodos , Dados de Sequência Molecular , Poli-Hidroxialcanoatos/biossíntese , Reação em Cadeia da Polimerase/métodosRESUMO
A direct relationship between biofilm formation and melanogenesis in Shewanella colwelliana with increased oyster recruitment is already established. Previously, S. colwelliana was grown in a newly patented biofilm-cultivation device, the conico-cylindrical flask (CCF), offering interchangeable hydrophobic/hydrophilic surfaces. Melanization was enhanced when S. colwelliana was cultivated in a hydrophobic vessel compared with a hydrophilic vessel. In the present study, melanogenesis in the CCF was positively correlated with increased architectural parameters of the biofilm (mean thickness and biovolume obtained by confocal laser scanning microscopy) and melanin gene (melA) expression observed by densitometry. Niche intertidal conditions were mimicked in a process operated in an ultra-low-speed rotating disk bioreactor, which demonstrated enhanced biofilm formation, melanogenesis, exopolysaccharide synthesis and melA gene expression compared with a process where 12-h periodic immersion and emersion was prevented. The wettability properties of the settling plane as well as intermittent wetting and drying, which influenced biofilm formation and melA expression, may affect oyster settlement in nature.
Assuntos
Biofilmes/crescimento & desenvolvimento , Melaninas/biossíntese , Shewanella/crescimento & desenvolvimento , Animais , Reatores Biológicos , Interações Hidrofóbicas e Hidrofílicas , Microscopia Confocal , Ostreidae , Shewanella/metabolismo , MolhabilidadeRESUMO
Haloferax mediterranei has potential for economical industrial-scale production of polyhydroxyalkanoate (PHA) as it can utilize cheap carbon sources, has capacity for nonsterile cultivation and allows simple product recovery. Molasses-based Indian distilleries are converting themselves to cereal-based distilleries. Waste stillage (14 l) of rice-based ethanol industry was used for the production of PHA by H. mediterranei in the simple plug-flow reactor configuration of the activated sludge process. Cells utilized stillage and accumulated 63 ± 3 % PHA of dry cell weight and produced 13.12 ± 0.05 g PHA/l. The product yield coefficient was 0.27 while 0.14 g/l h volumetric productivity was reached. Simultaneous lowering of 5-day biochemical oxygen demand and chemical oxygen demand values of stillage by 82 % was attained. The biopolymer was characterized as poly-3-(hydroxybutyrate-co-17.9 mol%-hydroxyvalerate) (PHBV). Directional properties of decanoic acid jointly with temperature-dependent water solubility in decanoic acid were employed for two-step desalination of the spent stillage medium in a cylindrical baffled-tank with an immersed heater and a stirrer holding axial and radial impellers. 99.3 % of the medium salts were recovered and re-used for PHA production. The cost of PHBV was estimated as US$2.05/kg when the annual production was simulated as 1890 tons. Desalination contributed maximally to the overall cost. Technology and cost-analysis demonstrate that PHA production integrated with ethanol manufacture is feasible in India. This study could be the basis for construction of a pilot plant.
Assuntos
Biotecnologia/economia , Biotecnologia/métodos , Haloferax mediterranei/genética , Haloferax mediterranei/metabolismo , Resíduos Industriais , Oryza/metabolismo , Poliésteres/metabolismo , Etanol/metabolismo , ÍndiaRESUMO
Haloferax mediterranei holds promise for competitive industrial-scale production of polyhydroxyalkanoate (PHA) because cheap carbon sources can be used thus lowering production costs. Although high salt concentration in production medium permits a non-sterile, low-cost process, salt disposal after process completion is a problem as current environmental standards do not allow total dissolved solids (TDS) above 2000 mg/l in discharge water. As the first objective of this work, the waste product of rice-based ethanol industry, stillage, was used for the production of PHA by H. mediterranei in shake flasks. Utilization of raw stillage led to 71 ± 2% (of dry cell weight) PHA accumulation and 16.42 ± 0.02 g/l PHA production. The product yield coefficient was 0.35 while 0.17 g/l h volumetric productivity was attained. Simultaneous reduction of BOD5 and COD values of stillage by 83% was accomplished. The PHA was isolated by osmotic lysis of cells, purification by sodium dodecyl sulfate and organic solvents. The biopolymer was identified as poly-3-(hydroxybutyrate-co-15.4 mol%-hydroxyvalerate) (PHBV). This first report on utilization of rice-based ethanol stillage for PHBV production by H. mediterranei is currently the most cost effective. As the second objective, directional properties of decanoic acid together with temperature dependence of water solubility in decanoic acid were applied for two-stage desalination of the spent stillage medium. We report for the first time, recovery and re-use of 96% of the medium salts for PHA production thus removing the major bottleneck in the potential application of H. mediterranei for industrial production of PHBV. Final discharge water had TDS content of 670 mg/l.
Assuntos
Biotecnologia/métodos , Etanol/metabolismo , Haloferax mediterranei/metabolismo , Oryza/química , Poliésteres/metabolismo , Fermentação , Haloferax mediterranei/crescimento & desenvolvimento , Poliésteres/química , Sais/metabolismoRESUMO
BACKGROUND: A major barrier to effective treatment of glioblastoma multiforme (GBM) is the invasion of glioma cells into the brain parenchyma rendering local therapies such as surgery and radiation therapy ineffective. GBM patients with such highly invasive and infiltrative tumors have poor prognosis with a median survival time of only about a year. However, the mechanisms leading to increased cell migration, invasion and diffused behavior of glioma cells are still poorly understood. METHODS: In the current study, we applied quantitative proteomics for the identification of differentially expressed proteins in GBMs as compared to non-malignant brain tissues. RESULTS: Our study led to the identification of 23 proteins showing overexpression in GBM; these include membrane proteins, moesin and CD44. The results were verified using Western blotting and immunohistochemistry in independent set of GBM and non-malignant brain tissues. Both GBM tissues and glioma cell lines (U87 / U373) demonstrated membranous expression of moesin and CD44, as revealed by immunohistochemistry and immunofluorescence, respectively. Notably, glioma cells transfected with moesin siRNA displayed reduced migration and invasion on treatment with hyaluronan (HA), an important component of the extracellular matrix in GBM. CD44, a transmembrane glycoprotein, acts as a major receptor for hyaluronan (HA). Using co-immunoprecipitation assays, we further demonstrated that moesin interacts with CD44 in glioma cells only after treatment with HA; this implicates a novel role of moesin in HA-CD44 signaling in gliomas. CONCLUSIONS: Our results suggest that development of inhibitors which interfere with CD44-moesin interactions may open a new avenue in the future to mitigate cellular migration in gliomas.
Assuntos
Movimento Celular/efeitos dos fármacos , Glioblastoma/metabolismo , Ácido Hialurônico/farmacologia , Proteínas dos Microfilamentos/metabolismo , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Glioblastoma/genética , Humanos , Receptores de Hialuronatos/genética , Receptores de Hialuronatos/metabolismo , Proteínas dos Microfilamentos/genética , Ligação Proteica/efeitos dos fármacos , Proteoma , ProteômicaRESUMO
High resolution compressed sensing hyperpolarized (13)C magnetic resonance spectroscopic imaging was applied in orthotopic human glioblastoma xenografts for quantitative assessment of spatial variations in (13)C metabolic profiles and comparison with histopathology. A new compressed sensing sampling design with a factor of 3.72 acceleration was implemented to enable a factor of 4 increase in spatial resolution. Compressed sensing 3D (13)C magnetic resonance spectroscopic imaging data were acquired from a phantom and 10 tumor-bearing rats following injection of hyperpolarized [1-(13)C]-pyruvate using a 3T scanner. The (13)C metabolic profiles were compared with hematoxylin and eosin staining and carbonic anhydrase 9 staining. The high-resolution compressed sensing (13)C magnetic resonance spectroscopic imaging data enabled the differentiation of distinct (13)C metabolite patterns within abnormal tissues with high specificity in similar scan times compared to the fully sampled method. The results from pathology confirmed the different characteristics of (13)C metabolic profiles between viable, non-necrotic, nonhypoxic tumor, and necrotic, hypoxic tissue.
Assuntos
Biomarcadores Tumorais/metabolismo , Neoplasias Encefálicas/metabolismo , Compressão de Dados/métodos , Glioblastoma/metabolismo , Imageamento por Ressonância Magnética/métodos , Espectroscopia de Ressonância Magnética/métodos , Proteínas de Neoplasias/metabolismo , Animais , Isótopos de Carbono , Linhagem Celular Tumoral , Humanos , Imageamento Tridimensional/métodos , Masculino , Imagem Molecular/métodos , Ratos , Ratos Nus , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Distribuição TecidualRESUMO
The aims of the investigation were to ascertain if surface attachment of Cunninghamella elegans and niche intertidal conditions provided in a bioreactor influenced biotransformation of fluoranthene by C. elegans. A newly designed polymethylmethacrylate (PMMA) conico-cylindrical flask (CCF) holding eight equidistantly spaced rectangular strips mounted radially on a circular disc allowed comparison of fluoranthene biotransformation between CCFs with a hydrophobic surface (PMMA-CCF) and a hydrophilic glass surface (GS-CCF) and a 500-ml Erlenmeyer flask (EF). Fluoranthene biotransformation was higher by 22-fold, biofilm growth was higher by 3-fold, and cytochrome P450 gene expression was higher by 2.1-fold when C. elegans was cultivated with 2% inoculum as biofilm culture in PMMA-CCF compared to planktonic culture in EF. Biotransformation was enhanced by 7-fold with 10% inoculum. The temporal pattern of biofilm progression based on three-channel fluorescence detection by confocal laser scanning microscopy demonstrated well-developed, stable biofilm with greater colocalization of fluoranthene within extracellular polymeric substances and filaments of the biofilm grown on PMMA in contrast to a glass surface. A bioreactor with discs rotating at 2 revolutions per day affording 6-hourly emersion and immersion mimicked the niche intertidal habitat of C. elegans and supported biofilm formation and transformation of fluoranthene. The amount of transformed metabolite was 3.5-fold, biofilm growth was 3-fold, and cytochrome P450 gene expression was 1.9-fold higher in the process mimicking the intertidal conditions than in a submerged process without disc rotation. In the CCF and reactor, where biofilm formation was comparatively greater, higher concentration of exopolysaccharides allowed increased mobilization of fluoranthene within the biofilm with consequential higher gene expression leading to enhanced volumetric productivity.
Assuntos
Biofilmes/crescimento & desenvolvimento , Cunninghamella/metabolismo , Fluorenos/metabolismo , Reatores Biológicos , Biotransformação , Cunninghamella/enzimologia , Cunninghamella/isolamento & purificação , Cunninghamella/fisiologia , Sistema Enzimático do Citocromo P-450/metabolismoRESUMO
The present study is an attempt to assess the radiogenic quality of groundwater on the basis of gross α, gross ß and tritium (3H or H-3) activities in the Bakreswar-Tantloi geothermal region of Chotanagpur Plateau, West Bengal and Jharkhand, India. The aforesaid parameters in groundwater samples were measured using liquid scintillation counting triple to double coincidence ratio (LSC-TDCR) technique. Groundwater samples collected from Bakreswar-Tantloi geothermal region show gross α activities from below the minimum detectable activity (BMDA) to 0.5 ± 0.05 Bq/L, gross ß activities from BMDA to 0.2 ± 0.01 Bq/L and H-3 activities from BMDA to 63.42 Bq/L. The average gross α, gross ß and H-3 activities are also within the permissible limits prescribed by the World Health Organization (WHO). Though the annual effective doses in some samples were higher than the reference dose level of 0.1 mSv, the overall result suggests that the groundwater in the Bakreswar-Tantloi geothermal region is radiologically safe considering the radionuclides covered in this study.