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Background: Cervical cancer is a public health problem in India due to weak national screening policy compounded by lack of resources including scarcity of trained personnel to carry out community-based screening program. Para medical professionals (PMPs) are closely related to women in local communities. Hence, training PMPs by incorporating novel technology and reduced time duration to achieve adequate competence in screening is an area underutilized and needs to be explored. Materials and methods: A pilot cross sectional analytical study was conducted at a tertiary referral cancer center using a shorter version of educational intervention of 2 weeks duration (EI2W) involving PMPs. Pre- and post-training assessment of knowledge, attitude, and practice (KAP) was done using questionnaires consisting of 5 domains viz. awareness of cervical cancer, awareness of cervical pre-cancer, practical screening methodology (practice oriented), data management and aspects of human papilloma virus (HPV). Wilcoxon signed-rank test was used for comparison and the degree of change was measured using analysis of covariance (ANCOVA). A p value of <0.05 was considered significant. Results: 118 PMPs were included. There was a significant improvement in scores of all domains (except cervical pre-cancer domain), following introduction of EI2W. Knowledge scores, post EI2W was better in Auxiliary Nurse Midwives (ANMs) than other participants. Awareness regarding cervical cancer was higher with more years of experience. The KAP analysis showed excellent interrater reliability in the practice 0.726 (0.649-0.792) followed by knowledge domain 0.711 (0.626-0.783). Conclusion: EI2W was effective in significantly improving the competence of PMPs, thus reducing human resource constraints in cervical cancer prevention and elimination.
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BACKGROUND: The main goal of glioma surgery is to remove the maximum amount of tumor without worsening the patient's neurological condition. Intraoperative ultrasound (US) imaging technologies (2D and 3D) are available to assist surgeons, providing real-time updates. Considering additional time, personnel, and cost, we investigate if comparable outcomes can be achieved using basic (2D) and advanced (3D) technology. OBJECTIVE: We propose predictive models for (i) glioma tumor resectability (ii) surgical outcome, and (iii) a model to predict the outcome of surgery aided with a particular ultrasound and compare outcomes between 2D and 3D US. METHODOLOGY: We used real-world surgery data from a tertiary cancer centre. Three groups of cases were analyzed (2D US used, 3D US used, and no US used during resection). The data analysis uses hypothesis testing, bootstrap sampling, and logistic regression. RESULTS: The preoperatively anticipated extent of tumor removal correlated with the postoperative MRI measurement of tumor removal for US-supported surgery (p=0.01) but not for no US-supported surgeries (p = 0.13). A combination of delineation, eloquence, and the multifocal/multicentric nature of the tumor effectively predicted resectability. The eventual outcome of surgery (actual extent of resection achieved) can be predicted by prior treatment status, delineation, eloquence, and satellite nodules. Based on our prediction model (training set of 350 cases and test of 40 cases of US-guided surgeries), we identify some cases where 3D US seems to offer superior EORs. CONCLUSION: The resectability of glioma tumors is crucial in determining surgical objectives, and the type of ultrasound used as support impacts tumor removal. The findings in this study aid informed decision-making and optimize imaging technology usage, providing a decision flow for selecting ultrasound based on tumor characteristics.
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Neoplasias Encefálicas , Glioma , Humanos , Neoplasias Encefálicas/diagnóstico por imagen , Neoplasias Encefálicas/cirugía , Glioma/diagnóstico por imagen , Glioma/cirugía , Glioma/patología , Imagen por Resonancia Magnética/métodosRESUMEN
The ever-increasing global energy demand has led world towards negative repercussions such as depletion of fossil fuels, pollution, global warming and climate change. Designing microbial cell factories for the sustainable production of biofuels is therefore an active area of research. Different yeast cells have been successfully engineered using synthetic biology and metabolic engineering approaches for the production of various biofuels. In the present article, recent advancements in genetic engineering strategies for production of bioalcohols, isoprenoid-based biofuels and biodiesels in different yeast chassis designs are reviewed, along with challenges that must be overcome for efficient and high titre production of biofuels.
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Biocombustibles , Saccharomyces cerevisiae , Saccharomyces cerevisiae/metabolismo , Ingeniería Metabólica , Redes y Vías Metabólicas , Terpenos/metabolismoRESUMEN
BACKGROUND: Gliomas are among the most typical brain tumors tackled by neurosurgeons. During navigation for surgery of glioma brain tumors, preoperatively acquired static images may not be accurate due to shifts. Surgeons use intraoperative imaging technologies (2-Dimensional and navigated 3-Dimensional ultrasound) to assess and guide resections. This paper aims to precisely capture the importance of preoperative parameters to decide which type of ultrasound to be used for a particular surgery. METHODS: This paper proposes two bagging algorithms considering base classifier logistic regression and random forest. These algorithms are trained on different subsets of the original data set. The goodness of fit of Logistic regression-based bagging algorithms is established using hypothesis testing. Furthermore, the performance measures for random-forest-based bagging algorithms used are AUC under ROC and AUC under the precision-recall curve. We also present a composite model without compromising the explainability of the models. RESULTS: These models were trained on the data of 350 patients who have undergone brain surgery from 2015 to 2020. The hypothesis test shows that a single parameter is sufficient instead of all three dimensions related to the tumor ([Formula: see text]). We observed that the choice of intraoperative ultrasound depends on the surgeon making a choice, and years of experience of the surgeon could be a surrogate for this dependence. CONCLUSION: This study suggests that neurosurgeons may not need to focus on a large set of preoperative parameters in order to decide on ultrasound. Moreover, it personalizes the use of a particular ultrasound option in surgery. This approach could potentially lead to better resource management and help healthcare institutions improve their decisions to make the surgery more effective.
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Neoplasias Encefálicas , Glioma , Humanos , Ultrasonografía/métodos , Glioma/diagnóstico por imagen , Glioma/cirugía , Neoplasias Encefálicas/diagnóstico por imagen , Neoplasias Encefálicas/cirugía , Neoplasias Encefálicas/patología , AlgoritmosRESUMEN
Introduction: The coronavirus disease 2019 (COVID-19) pandemic has caused significant global disruption, especially for chronic care like hemodialysis treatments. Approximately 10,000 end-stage kidney disease (ESKD) patients are receiving maintenance hemodialysis (MHD) at 174 dialysis centers in Greater Mumbai. Because of the fear of transmission of infection and inability to isolate patients in dialysis centers, chronic hemodialysis care was disrupted for COVID-19-infected patients. Hence, we embarked on a citywide initiative to ensure uninterrupted dialysis for these patients. Materials and Methods: The Municipal Corporation of Greater Mumbai (MCGM) designated 23 hemodialysis facilities as COVID-positive centers, two as COVID-suspect centers, and the rest continued as COVID-negative centers to avoid transmission of infection and continuation of chronic hemodialysis treatment. Nephrologists and engineers of the city developed a web-based-portal so that information about the availability of dialysis slots for COVID-infected patients was easily available in real time to all those providing care to chronic hemodialysis patients. Results: The portal became operational on May 20, 2020, and as of December 31, 2020, has enrolled 1,418 COVID-positive ESKD patients. This initiative has helped 97% of enrolled COVID-infected ESKD patients to secure a dialysis slot within 48 hours. The portal also tracked outcomes and as of December 31, 2020, 370 (27%) patients died, 960 patients recovered, and 88 patients still had an active infection. Conclusions: The portal aided the timely and smooth transfer of COVID-19-positive ESKD patients to designated facilities, thus averting mortality arising from delayed or denied dialysis. Additionally, the portal also documented the natural history of the COVID-19 pandemic in the city and provided information on the overall incidence and outcomes. This aided the city administration in the projected resource needs to handle the pandemic.
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Extracellular signals are reported to mediate chemical cross-talk among pelagic microbes, including microalgal prey and predators. Water-soluble mediator compounds play a crucial role in extracellular communication which is vital for prey recognition, attraction, capture, and predator deterrence. A range of exo-metabolites including oxylipins and vitamins are released by prey in response to grazing stress. The temporal dynamics of such exo-metabolites largely remains unknown, especially in large-scale cultivation of microalgae such as closed or open ponds. In open ponds, infestation of predators is almost inevitable but highly undesirable due to the imminent threat of culture collapse. The early production of exo-metabolites emitted by microalgal prey in response to predator attack could be leveraged as diagnostic markers of possible culture collapse. This study uses an untargeted approach for temporal profiling of Dunaliella tertiolecta-specific exo-metabolites under grazing pressure from Oxyrrhis marina. We report 24 putatively identified metabolites, belonging to various classes such as short peptides, lipids, indole-derivatives, and free amino acids, as potential markers of grazing-mediated stress. In addition, this study outlines a clear methodology for screening of exo-metabolites in marine algal samples, the analysis of which is frequently hindered by high salt concentrations. In future, a chemistry-based targeted detection of these metabolites could enable a quick and on-site screening of predators in microalgal cultures.
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Chlorophyceae , Dinoflagelados , Microalgas , Cromatografía Liquida , Dinoflagelados/metabolismo , Microalgas/metabolismo , Espectrometría de Masas en TándemRESUMEN
Salinity is one of the significant factors that affect growth and cellular metabolism, including photosynthesis and lipid accumulation, in microalgae and higher plants. Microchloropsis gaditana CCMP526 can acclimatize to different salinity levels by accumulating compatible solutes, carbohydrates, and lipids as energy storage molecules. We used proteomics to understand the molecular basis for acclimation of M. gaditana to increased salinity levels [55 and 100 PSU (practical salinity unit)]. Correspondence analysis was used for the identification of salinity-responsive proteins (SRPs). The highest number of salinity-induced proteins was observed in 100 PSU. Gene ontology enrichment analysis revealed a separate path of acclimation for cells exposed to 55 and 100 PSU. Osmolyte and lipid biosynthesis were upregulated in hypersaline conditions. Concomitantly, lipid oxidation pathways were also upregulated in hypersaline conditions, providing acetyl-CoA for energy metabolism through the tricarboxylic acid cycle. Carbon fixation and photosynthesis were tightly regulated, while chlorophyll biosynthesis was affected in hypersaline conditions. Importantly, temporal proteome analysis of salinity-induced M. gaditana revealed vital SRPs which could be used for engineering salinity resilient microalgal strains for improved productivity in hypersaline culture conditions.
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BACKGROUND: Increased acetylcholinesterase (AChE) activity on frozen sections of rectal mucosal biopsies accurately diagnoses Hirschsprung disease (HD). But the quest for a biomarker in blood as a screening test prompts one to look for AChE in blood and study its role in HD diagnosis. AIM: To develop a low-cost reliable method to estimate the AChE activity in plasma and red blood cells (RBCs) in normal children (control) and study its role in HD (test). MATERIALS AND METHODS: Optimized method derived after modifying and standardizing known AChE assay protocols for blood were employed on 30 controls to define the AChE cut-off range, on 40 suspected HD cases to categorize them as HD/non-HD based on cut-off values and later compared with gold standard tissue AChE histochemistry of rectal mucosal biopsies. RESULTS: An optimal in-house modified methods of Ellman's was found best suited to analyze plasma AChE activity, method by Wilson and Henderson was optimal for extraction and AChE estimation in RBCs. AChE levels (controls) obtained were 1.03 ± 0.31 U/mL and 5.17 ± 1.52 U/mL in plasma and RBCs, respectively while the plasma AChE was 1.35 ± 0.84 U/mL (HD) and 1.62 ± 0.85 U/mL (non-HD) while RBC AChE was 4.29 ± 3.2 U/mL (HD) and 6.48 ± 4.31 U/mL (non-HD). Sensitivity was 66.67% and 55.56%, specificity was 22.73% and 45.45%, and an accuracy rate of 42.5% and 50% for plasma and RBC, respectively. CONCLUSIONS: Mutually exclusive AChE activity range identified for test blood samples overlapped with the normal and hence, not considered a diagnostic tool for HD.
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Acetilcolinesterasa/análisis , Acetilcolinesterasa/sangre , Eritrocitos/enzimología , Enfermedad de Hirschsprung/sangre , Enfermedad de Hirschsprung/diagnóstico , Recto/enzimología , Acetilcolinesterasa/metabolismo , Biomarcadores/análisis , Biomarcadores/sangre , Biopsia , Niño , Preescolar , Método Doble Ciego , Tránsito Gastrointestinal/fisiología , Enfermedad de Hirschsprung/patología , Histocitoquímica/métodos , Humanos , India , Membrana Mucosa/enzimología , Estudios Prospectivos , Sensibilidad y EspecificidadRESUMEN
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA-dependent RNA polymerase (RdRp) has been identified to be a mutation hot spot, with the P323L mutation being commonly observed in viral genomes isolated from North America. RdRp forms a complex with nonstructural proteins nsp7 and nsp8 to form the minimal replication/transcription machinery required for genome replication. As mutations in RdRp may affect formation of the RdRp-nsp7-nsp8 supercomplex, we analyzed viral genomes to identify mutations in nsp7 and nsp8 protein sequences. Based on in silico analysis of predicted structures of the supercomplex comprising of native and mutated proteins, we demonstrate that specific mutations in nsp7 and nsp8 proteins may have a role in stabilization of the replication/transcription complex.
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ARN Polimerasa Dependiente de ARN de Coronavirus/genética , SARS-CoV-2/fisiología , Proteínas no Estructurales Virales/genética , Compartimentos de Replicación Viral/química , Secuencia de Aminoácidos , Simulación por Computador , ARN Polimerasa Dependiente de ARN de Coronavirus/química , ARN Polimerasa Dependiente de ARN de Coronavirus/metabolismo , Genoma Viral , Humanos , Modelos Moleculares , Mutación , Estabilidad Proteica , SARS-CoV-2/química , SARS-CoV-2/genética , Proteínas no Estructurales Virales/química , Proteínas no Estructurales Virales/metabolismo , Compartimentos de Replicación Viral/metabolismoRESUMEN
The microbial production of industrial enzymes requires a large number of complex biochemical steps for purification which increases their production cost. Additionally, poor thermo-stability of the purified enzymes under the operational conditions along with the challenges in their recovery and subsequent reuse, limit their usage in an industrial bioprocess. Surface display of heterologous enzymes on bacterial cells appear to be a suitable alternative. Bacillus subtilis, the most well characterized Gram-positive bacterium, is being increasingly studied as a host for surface display. We displayed a glucose-tolerant ß-glucosidase (UnBgl1A) on the surface of B. subtilis cells using CWBb as the anchor protein. These cells displaying UnBgl1A (SD-01) were directly employed for biocatalysis without cell lysis and enzyme purification. The SD-01 cells elicited â¼2 times more catalytic activity compared to the cells expressing the enzyme intracellularly (IN-01). The displayed enzyme and the purified enzyme elucidated similar glucose tolerance (IC50 â¼0.9â¯M glucose), temperature optima (â¼50⯰C), and pH optima (â¼6.0). The surface displayed UnBgl1A retained â¼50% activity after 4â¯h when stored at 50⯰C whereas the purified UnBgl1A lost all its activity by the 4th hour. Additionally, the SD-01 cells could be efficiently reused for 3 sets of reactions. Further, supplementation of a cellulase cocktail with the cells of the SD-01 strain resulted in â¼2 times more glucose release from sugarcane bagasse compared to supplementation with the purified UnBgl1A. Therefore, displaying enzymes on the B. subtilis cell surface could be an attractive platform for the commercial production of industrial enzymes.
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Bacillus subtilis/enzimología , Glucosa/metabolismo , beta-Glucosidasa/metabolismo , Bacillus subtilis/genética , Proteínas Bacterianas/metabolismo , Biocatálisis , Celulosa , Clonación Molecular , Estabilidad de Enzimas , Concentración de Iones de Hidrógeno , TemperaturaRESUMEN
Evaporation from culture ponds and raceways can subject algae to hypersalinity stress, and this is exacerbated by global warming. We investigated the effect of salinity on a marine microalga, Microchloropsis gaditana, which is of industrial significance because of its high lipid-accumulating capability. Both short-term (hours) and medium-term (days) effects of salinity were studied across various salinities (37.5, 55, 70 and 100â¯PSU). Salinity above 55â¯PSU suppressed cell growth and specific growth rate was significantly reduced at 100â¯PSU. Photosynthesis (Fv/Fm, rETRmax and Ik) was severely affected at high salinity conditions. Total carbohydrate per cell increasedâ¯â¼1.7-fold after 24â¯h, which is consistent with previous findings that salinity induces osmolyte production to counter osmotic shock. In addition, accumulation of lipid increased byâ¯â¼4.6-fold in response to salinity. Our findings indicate a possible mechanism of acclimation to salinity, opening up new frontiers for osmolytes in pharmacological and cosmetics applications.
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Fotosíntesis , Salinidad , Aclimatación , Microalgas , Presión OsmóticaRESUMEN
Proteomics is a crucial postgenomic biotechnology for functional and systems scale analyses in cell and integrative biology, not to mention clinical and precision medicine research. However, a fundamental requirement for an accurate examination of the protein complement of cells is an efficient method for extracting the proteins. This study reports on the evaluation of three protein extraction methods: trichloroacetic acid (TCA)-acetone, phenol, and TRIzol, in the eustigmatophyte alga Microchloropsis gaditana CCMP526 for proteomic analysis. M. gaditana is a potential candidate for algal-based biofuels. This industrially important strain is also rich in dietary oil and pigments and is used as feed in the aquaculture industry. Liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based proteomic analysis was performed for proteins obtained using the three extraction methods and their effects were examined by the abundance ratio. Protein yield was higher using the TCA-acetone and phenol methods than with the TRIzol method. The TCA method was superior than other methods examined here in terms of protein coverage and abundance. Subcellular localization of the identified proteins revealed no significant difference among the extraction methods. Importantly, each method revealed a unique set of proteins. To the best of our knowledge, this is the first report on evaluation of protein extraction methods for the proteomic analysis of M. gaditana CCMP526. These observations underscore the importance of using multiple protein extraction methods for comprehensive proteome coverage, as the field of proteomics edges toward diverse applications in biofuels, aquaculture industry, marine biology, and agriculture.
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Proteínas/aislamiento & purificación , Proteoma/aislamiento & purificación , Proteómica/métodos , Estramenopilos/química , Acetona/química , Biocombustibles , Guanidinas/química , Fenol/química , Fenoles/química , Ácido Tricloroacético/químicaRESUMEN
Meningiomas are one of the most common tumors of the Central nervous system (CNS). This study aims to identify the autoantibody biomarkers in meningiomas using high-density human proteome arrays (~17,000 full-length recombinant human proteins). Screening of sera from 15 unaffected healthy individuals, 10 individuals with meningioma grade I and 5 with meningioma grade II was performed. This comprehensive proteomics based investigation revealed the dysregulation of 489 and 104 proteins in grades I and II of meningioma, respectively, along with the enrichment of several signalling pathways, which might play a crucial role in the manifestation of the disease. Autoantibody targets like IGHG4, CRYM, EFCAB2, STAT6, HDAC7A and CCNB1 were significantly dysregulated across both the grades. Further, we compared this to the tissue proteome and gene expression profile from GEO database. Previously reported upregulated proteins from meningioma tissue-based proteomics obtained from high-resolution mass spectrometry demonstrated an aggravated autoimmune response, emphasizing the clinical relevance of these targets. Some of these targets like SELENBP1 were tested for their presence in tumor tissue using immunoblotting. In the light of highly invasive diagnostic modalities employed to diagnose CNS tumors like meningioma, these autoantibody markers offer a minimally invasive diagnostic platform which could be pursued further for clinical translation.
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Current momentum of microalgal research rests extensively in tapping the potential of multi-omics methodologies in regard to sustainable biofuels. Microalgal biomass is fermented to bioethanol; while lipids, particularly triacylglycerides (TAGs), are transesterified to biodiesels. Biodiesel has emerged as an ideal biofuel candidate; hence, its commercialization and use are increasingly being emphasized. Abiotic stresses exaggerate TAG accumulation, but the precise mechanisms are yet to be known. More recently, comprehensive multi-omics studies in microalgae have emerged from the biofuel perspective. Genomics and transcriptomics of microalgae have provided crucial leads and basic understanding toward lipid biosynthesis. Proteomics and metabolomics are now complementing "algal omics" and offer precise functional insights into the attendant static and dynamic physiological contexts. Indeed, the field has progressed from shotgun to targeted approaches. Notably, targeted proteomics studies in microalga are not yet reported. Several multi-omics tools and technologies that may be used to dig deeper into the microalgal physiology are examined and highlighted in this review. The article therefore aims to both introduce various available high-throughput biotechnologies and applications of "omics" in microalgae, and enlists a compendium of the emerging cutting edge literature. We suggest that a strategic and thoughtful combination of data streams from different omics platforms can provide a system-wide overview. The algal omics warrants closer attention in the future, with a view to technical, economic, and societal impacts that are anticipated in the current postgenomics era.
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Biocombustibles , Biotecnología/métodos , Microalgas/metabolismo , Genómica , Metabolómica , Microalgas/genética , ProteómicaRESUMEN
While non-infectious etiologies like chemical irritants are rare causes of epidemics of peritonitis, this possibility should be considered when one encounters an unusual clustering of peritonitis cases. We describe here an epidemic of chemical peritonitis at our center.
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Soluciones para Diálisis/efectos adversos , Epidemias , Fallo Renal Crónico/terapia , Diálisis Peritoneal Ambulatoria Continua/efectos adversos , Peritonitis/inducido químicamente , Peritonitis/epidemiología , Adolescente , Adulto , Anciano , Femenino , Humanos , India/epidemiología , Masculino , Persona de Mediana Edad , Peritonitis/diagnóstico , Adulto JovenRESUMEN
In the present study, we explored the extent to which inaccuracies inherent in homology models of the transmembrane helical cores of G protein-coupled receptors (GPCRs) can impact loop prediction. We demonstrate that loop prediction in homology models is much more difficult than loop reconstruction in crystal structures because of the imprecise positioning of loop anchors. Deriving information from 17 recently available GPCR crystal structures, we estimated all of the possible errors that could occur in loop anchors as the result of comparative modeling. Subsequently, we performed an exhaustive analysis to decipher the effect of these errors on loop modeling using ICM High Precision Sampling. The influence of the presence of other extracellular loops was also explored. Our results reveal that the error space of modeled loop residues is much larger than that of the anchor residues, although modeling a particular extracellular loop in the presence of other extracellular loops provides constraints that help in predicting near-native loop conformations observed in crystal structures. This implies that errors in loop anchor positions introduce increased uncertainty in the modeled loop coordinates. Therefore, for the success of any GPCR structure prediction algorithm, minimizing errors in the helical end points is likely to be critical for successful loop modeling.
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Modelos Moleculares , Receptores Acoplados a Proteínas G/química , Homología de Secuencia de Aminoácido , Secuencia de Aminoácidos , Cristalografía por Rayos X , Método de Montecarlo , Conformación Proteica en Hélice alfa , TermodinámicaRESUMEN
Escherichia coli cra null mutants have been reported in the literature to be impaired in biofilm formation. To develop E. coli biofilm-inhibiting agents for prevention and control of adherent behaviour, analogues of a natural Cra ligand, fructose-1,6-bisphosphate, were identified based on two-dimensional similarity to the natural ligand. Of the analogues identified, those belonging to the bisphosphonate class of drug molecules were selected for study, as these are approved for clinical use in humans and their safety has been established. Computational and in vitro studies with purified Cra protein showed that risedronate sodium interacted with residues in the fructose-1,6-bisphosphate-binding site. Using a quantitative biofilm assay, risedronate sodium, at a concentration of 300-400 µM, was found to decrease E. coli and Salmonella pullorum biofilm formation by >60 %. Risedronate drastically reduced the adherence of E. coli cells to a rubber Foley urinary catheter, demonstrating its utility in preventing the formation of biofilm communities on medical implant surfaces. The use of risedronate, either alone or in combination with other agents, to prevent the formation of biofilms on surfaces is a novel finding that can easily be translated into practical applications.
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Adhesión Bacteriana/efectos de los fármacos , Biopelículas/efectos de los fármacos , Escherichia coli/efectos de los fármacos , Ácido Risedrónico/farmacología , Proteínas Bacterianas/metabolismo , Clonación Molecular , Fructosadifosfatos/química , Eliminación de Gen , Proteínas Represoras/metabolismo , Salmonella/efectos de los fármacos , Cateterismo UrinarioRESUMEN
The heterogeneity and poor prognosis associated with gliomas, makes biomarker identification imperative. Here, we report autoantibody signatures across various grades of glioma serum samples and sub-categories of glioblastoma multiforme using Human Proteome chips containing ~17000 full-length human proteins. The deduced sets of classifier proteins helped to distinguish Grade II, III and IV samples from the healthy subjects with 88, 89 and 94% sensitivity and 87, 100 and 73% specificity, respectively. Proteins namely, SNX1, EYA1, PQBP1 and IGHG1 showed dysregulation across various grades. Sub-classes of GBM, based on its proximity to the sub-ventricular zone, have been reported to have different prognostic outcomes. To this end, we identified dysregulation of NEDD9, a protein involved in cell migration, with probable prognostic potential. Another subcategory of patients where the IDH1 gene is mutated, are known to have better prognosis as compared to patients carrying the wild type gene. On a comparison of these two cohorts, we found STUB1 and YWHAH proteins dysregulated in Grade II glioma patients. In addition to common pathways associated with tumourigenesis, we found enrichment of immunoregulatory and cytoskeletal remodelling pathways, emphasizing the need to explore biochemical alterations arising due to autoimmune responses in glioma.
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Autoanticuerpos/sangre , Autoanticuerpos/inmunología , Glioma/sangre , Glioma/inmunología , Proteoma , Proteómica , Biomarcadores , Glioma/patología , Humanos , Clasificación del Tumor , Proteómica/métodosRESUMEN
The aim of the study was to increase production of (R)-PAC by altering carboligation activity of Pdc in Saccharomyces cerevisiae. Pdc1 activity was modified by over-expression as well as changing the rate of decarboxylation and carboligation by site specific mutation in Pdc1. Over-expression of mutant Pdc1 resulted in 50 ± 2.5% increase in levels of (R)-PAC in wild-type and further 30-40% in pdc null background. The combination of mutant Pdc1 in pdc null background was successfully evaluated for production of (R)-PAC at industrial scale. This is the first report of enhancing (R)-PAC product in yeast by recombinant technology with capability of commercial production.
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Piruvato Descarboxilasa/genética , Piruvato Descarboxilasa/metabolismo , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Descarboxilación/genética , Metanol/metabolismo , Mutación/genética , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismoRESUMEN
(R)-phenylacetylcarbinol or (R)-PAC is a pharmaceutical precursor of (1R, 2S) ephedrine and (1S, 2S) pseudoephedrine. Biotransformation of benzaldehyde and glucose by pyruvate decarboxylase produces (R)-PAC. This biotransformation suffers from toxicity of the substrate, product [(R)-PAC] and by-product (benzyl alcohol). In the present study, ionic liquid/aqueous biphasic system was employed to enhance (R)-PAC production. Fermented broth was the reaction medium in which Saccharomyces cerevisiae BY4741 was the source of pyruvate decarboxylase. Hydrophobic ionic liquid 1-butyl-3-methylimidazolium hexafluorophosphate ([BMIM][PF6]) was the non-aqueous phase in which toxic compounds reside. Biocompatibility of [BMIM][PF6] and adequate distribution coefficients of benzaldehyde, (R)-PAC and benzyl alcohol were determined. A Box-Behnken design and response surface methodology were used for the optimization of biotransformation variables in order to maximize (R)-PAC yield and productivity. The results showed higher (R)-PAC yield and productivity of â¼1.5-fold each in the biphasic biotransformation of phase volume ratio 0.05 as compared to the monophasic (conventional) biotransformation. Moreover, the level of major by-product benzyl alcohol was also 3.5-fold lower in biphasic biotransformation. [BMIM][PF6]/aqueous biphasic system is a new approach which could intensify the (R)-PAC production.