ABSTRACT
BACKGROUND: Spermatogenesis is a temperature-sensitive process, and elevation in temperature hampers this process quickly and significantly. We studied the molecular effects of testicular heating on piRNAs and gene expression in rat testicular germ cells. METHODS: We generated a cryptorchid rat model by displacing the testis from the scrotal sac (34 °C) to the abdominal area (37 °C) and sacrificed animals after 1 day, 3 days, and 5 days. Pachytene spermatocytes and round spermatids were purified using elutriation centrifugation and percoll gradient methods. We performed transcriptome sequencing in pachytene spermatocytes and round spermatids to identify differentially expressed piRNAs and their probable targets, i.e., TE transcripts and mRNAs. RESULTS: As a result of heat stress, we observed significant upregulation of piRNAs and TE transcripts in testicular germ cells. In addition to this, piRNA biogenesis machinery and heat shock proteins (Hsp70 and Hsp90 family members) were upregulated. mRNAs have also been proposed as targets for piRNAs; therefore, we shortlisted certain piRNA-mRNA pairs with an inverse relationship of expression. We observed that in testicular heat stress, the heat shock proteins go hand-in-hand with the upregulation of piRNA biogenesis machinery. The dysregulation of piRNAs in heat-stressed germ cells, increased ping-pong activity, and disturbed expression of piRNA target transcripts suggest a connection between piRNAs, mRNAs, and TE transcripts. CONCLUSIONS: In heat stress, piRNAs, piRNA machinery, and heat shock proteins are activated to deal with low levels of stress, which is followed by a rescue approach in prolonged stressaccompained by high TE activity to allow genetic mutations, perhaps for survival and adaptability.
Subject(s)
Heat-Shock Response , RNA, Small Interfering , Spermatids , Spermatocytes , Testis , Animals , Male , Spermatids/metabolism , Spermatocytes/metabolism , RNA, Small Interfering/genetics , Rats , Heat-Shock Response/genetics , Heat-Shock Response/physiology , Testis/metabolism , Spermatogenesis/genetics , Spermatogenesis/physiology , Pachytene Stage/genetics , Rats, Sprague-Dawley , Piwi-Interacting RNAABSTRACT
The present study focused on the isolation and identification of CP and TCP bacteria degrading bacteria from the rhizospheric zone of aromatic grasses i.e. palmarosa (Cymbopogon martinii (Roxb. Wats), lemongrass (Cymbopogon flexuosus) and vetiver (Chrysopogon zizaniodes (L.) Nash.). So that these isolates alone or in combination with the vegetation of aromatic grasses will be used to clean up CP-contaminated soils. The study also explored enzymatic activities, CO2 release, dechlorination potential, and degradation pathways of bacterial strains. A total of 53 CP-tolerant bacteria were isolated on their physical characteristics and their ability to degrade CP. The ten highly CP-tolerant isolates were Pseudomonas aeruginosa Pa608, three strains of Pseudomonas hibiscicola R4-721 from different rhizosphere, Enterococcus lectis PP2a, Pseudomonas monteilii NBFPALD_RAS131, Enterobacter cloacae L3, Stenotrophomonas maltophilia PEG-390, Escherichia coli ABRL132, and Escherichia coli O104:H4 strain FWSEC0009. The CO2 emission and phosphatase activities of the isolates varied from 3.1 to 8.6 µmol mL-1 and 12.3 to 31 µmol PNP h-1, respectively in the CP medium. The degradation kinetics of CP by these isolates followed a one-phase decay model with a dissipation rate ranging from 0.048 to 0.41 d-1 and a half-life of 1.7-14.3 days. The growth data fitted in the SGompertz equation showed a growth rate (K) of 0.21 ± 0.28 to 0.91 ± 0.33 d-1. The P. monteilii strain had a faster growth rate while E. coli ABRL132 had slower growth among the isolates. The rate of TCP accumulation calculated by the SGompertz equation was 0.21 ± 0.02 to 1.18 ± 0.19 d-1. The Pseudomonas monteilii showed a lower accumulation rate of TCP. Among these, four highly effective isolates were Pseudomonas aeruginosa Pa608, Pseudomonas monteilii NBFPALD_RAS131, Stenotrophomonas maltophilia PEG-390, and Pseudomonas hibiscicola R4-721. Illustrations of the degradation pathways indicated that the difference in metabolic pathways of each isolate was associated with their growth rate, phosphatase, dehydrogenase, oxidase, and dechlorination activities.
Subject(s)
Biodegradation, Environmental , Chlorpyrifos , Kinetics , Chlorpyrifos/metabolism , Bacteria/metabolism , Bacteria/classification , Soil Pollutants/metabolism , Soil Microbiology , Rhizosphere , Pseudomonas aeruginosa/metabolism , Pseudomonas/metabolism , Insecticides/metabolism , Carbon Dioxide/metabolism , Cymbopogon/metabolismABSTRACT
Stem cell therapies have emerged as a promising treatment strategy for various diseases characterized by ischemic injury such as ischemic stroke. Cell survival after transplantation remains a critical issue. We investigated the impact of oxidative stress, being typically present in ischemically challenged tissue, on human dental pulp stem cells (hDPSC) and human mesenchymal stem cells (hMSC). We used oxygen-glucose deprivation (OGD) to induce oxidative stress in hDPSC and hMSC. OGD-induced generation of O2â¢- or H2O2 enhanced autophagy by inducing the expression of activating molecule in BECN1-regulated autophagy protein 1 (Ambra1) and Beclin1 in both cell types. However, hDPSC and hMSC pre-conditioning using reactive oxygen species (ROS) scavengers significantly repressed the expression of Ambra1 and Beclin1 and inactivated autophagy. O2â¢- or H2O2 acted upstream of autophagy, and the mechanism was unidirectional. Furthermore, our findings revealed ROS-p38-Erk1/2 involvement. Pre-treatment with selective inhibitors of p38 and Erk1/2 pathways (SB202190 and PD98059) reversed OGD effects on the expression of Ambra1 and Beclin1, suggesting that these pathways induced oxidative stress-mediated autophagy. SIRT3 depletion was found to be associated with increased oxidative stress and activation of p38 and Erk1/2 MAPKs pathways. Global ROS inhibition by NAC or a combination of polyethylene glycol-superoxide dismutase (PEG-SOD) and polyethylene glycol-catalase (PEG-catalase) further confirmed that O2â¢- or H2O2 or a combination of both impacts stems cell viability by inducing autophagy. Furthermore, autophagy inhibition by 3-methyladenine (3-MA) significantly improved hDPSC viability. These findings contribute to a better understanding of post-transplantation hDPSC and hMSC death and may deduce strategies to minimize therapeutic cell loss under oxidative stress.
Subject(s)
Autophagy , Hydrogen Peroxide , Adaptor Proteins, Signal Transducing/metabolism , Apoptosis , Beclin-1/metabolism , Beclin-1/pharmacology , Cell Survival , Glucose/metabolism , Humans , Hydrogen Peroxide/pharmacology , Oxidative Stress , Oxygen/pharmacology , Reactive Oxygen Species/metabolism , Stem Cells/metabolismABSTRACT
BACKGROUND: Diacylglycerol kinase (DGK) regulates intracellular signaling and functions by converting diacylglycerol (DAG) into phosphatidic acid. We previously demonstrated that DGK inhibition attenuates airway smooth muscle (ASM) cell proliferation, however, the mechanisms mediating this effect are not well established. Given the capacity of protein kinase A (PKA) to effect inhibition of ASM cells growth in response to mitogens, we employed multiple molecular and pharmacological approaches to examine the putative role of PKA in the inhibition of mitogen-induced ASM cell proliferation by the small molecular DGK inhibitor I (DGK I). METHODS: We assayed cell proliferation using CyQUANT™ NF assay, protein expression and phosphorylation using immunoblotting, and prostaglandin E2 (PGE2) secretion by ELISA. ASM cells stably expressing GFP or PKI-GFP (PKA inhibitory peptide-GFP chimera) were stimulated with platelet-derived growth factor (PDGF), or PDGF + DGK I, and cell proliferation was assessed. RESULTS: DGK inhibition reduced ASM cell proliferation in cells expressing GFP, but not in cells expressing PKI-GFP. DGK inhibition increased cyclooxygenase II (COXII) expression and PGE2 secretion over time to promote PKA activation as demonstrated by increased phosphorylation of (PKA substrates) VASP and CREB. COXII expression and PKA activation were significantly decreased in cells pre-treated with pan-PKC (Bis I), MEK (U0126), or ERK2 (Vx11e) inhibitors suggesting a role for PKC and ERK in the COXII-PGE2-mediated activation of PKA signaling by DGK inhibition. CONCLUSIONS: Our study provides insight into the molecular pathway (DAG-PKC/ERK-COXII-PGE2-PKA) regulated by DGK in ASM cells and identifies DGK as a potential therapeutic target for mitigating ASM cell proliferation that contributes to airway remodeling in asthma.
Subject(s)
Cyclic AMP-Dependent Protein Kinases , Diacylglycerol Kinase , Diacylglycerol Kinase/metabolism , Diacylglycerol Kinase/pharmacology , Cyclic AMP-Dependent Protein Kinases/metabolism , Cyclic AMP-Dependent Protein Kinases/pharmacology , Cells, Cultured , Cell Proliferation , Myocytes, Smooth Muscle/metabolismABSTRACT
This study aims to assess the effects of inhaled nitric oxide (iNO) on oxygenation in the management of pulmonary hypertension (PH) secondary to arteriovenous malformations (AVMs) in neonates. This is a matched retrospective cohort study from January 1, 2013, to December 31, 2017. The European inhaled nitric oxide registry from 43 neonatal and pediatric ICUs in 13 countries across Europe was used to extract data. The target population was neonates treated with iNO for the management of PH. The cases (PH secondary to AVMs treated with iNO) were matched (1:4 ratio) to controls (PH without AVMs treated with iNO). The main outcome measure was the absolute change of oxygenation index (OI) from baseline to 60 min after starting iNO in cases and controls. The primary outcome of our study was that the mean absolute change in OI from baseline to after 60 min was higher among cases 10.7 (14), than in controls 6 (22.5), and was not statistically different between the groups. The secondary outcome variable - death before discharge - was found to be significantly higher in cases (55%) than in controls (8%). All the other variables for secondary outcome measures remained statistically insignificant. Conclusion: Infants with PH secondary to AVMs treated with iNO did not respond differently compared to those presented with PH without AVMs treated with iNO. Right ventricular dysfunction on echocardiography was higher in cases than controls (cases: 66.7% and controls: 28.6%) but was not statistically significant. What is Known: ⢠Arterioenous malformation (AVM) is a well-known cause of persistent pulmonary hypertension in newborns. Inhaled nitric oxide (iNO) is most commonly used as first-line therapy for pulmonary hypertension in newborns. ⢠Around 40-50% of vein of Galen malformations (VOGMs) are found to have congestive heart failure in the neonatal period. What is New: ⢠Neonates may present with an isolated PH of the newborn as the main feature of the VOGMs. A large proportion of cases with AVMs have been associated with right ventricular cardiac dysfunction. ⢠Results from one of the largest database registries in the world for iNO have been used to answer our research question.
Subject(s)
Arteriovenous Malformations , Hypertension, Pulmonary , Lung Diseases , Administration, Inhalation , Child , Humans , Hypertension, Pulmonary/drug therapy , Hypertension, Pulmonary/etiology , Infant, Newborn , Nitric Oxide/therapeutic use , Registries , Retrospective StudiesABSTRACT
The Par polarity complex, consisting of Par3, Par6 and atypical protein kinase C (aPKC), plays a crucial role in the establishment and maintenance of cell polarity. Although activation of aPKC is critical for polarity, how this is achieved is unclear. The developing zebrafish epidermis, along with its apical actin-based projections, called microridges, offers a genetically tractable system for unraveling the mechanisms of the cell polarity control. The zebrafish aPKC regulates elongation of microridges by controlling levels of apical Lgl, which acts as a pro-elongation factor. Here, we show that the nucleoporin Nup358 (also known as RanBP2) - a component of the nuclear pore complex and a part of cytoplasmic annulate lamellae (AL) - SUMOylates zebrafish aPKC. Nup358-mediated SUMOylation controls aPKC activity to regulate Lgl-dependent microridge elongation. Our data further suggest that cytoplasmic AL structures are the possible site for Nup358-mediated aPKC SUMOylation. We have unraveled a hitherto unappreciated contribution of Nup358-mediated aPKC SUMOylation in cell polarity regulation.This article has an associated First Person interview with the first author of the paper.
Subject(s)
Cell Polarity/physiology , Epidermal Cells/metabolism , Molecular Chaperones/metabolism , Nuclear Pore Complex Proteins/metabolism , Zebrafish/metabolism , Actins/metabolism , Animals , Epidermis/metabolism , Epithelial Cells/metabolism , Molecular Chaperones/genetics , Nuclear Pore Complex Proteins/genetics , Zebrafish/embryology , Zebrafish/geneticsABSTRACT
BACKGROUND: The most common pathological cause of abnormal vaginal discharge in reproductive-aged women is bacterial vaginosis (BV). Amsel's criteria and Nugent scoring systems are commonly employed approaches for the diagnosis of BV. Despite the Nugent scoring system being the gold standard method for diagnosing BV, Amsel's criteria are generally preferred in clinical setup owing to the fact Nugent scoring requires considerable time and expert microscopist. This study was conducted to determine the diagnostic value of Amsel's criteria by comparing it with the Nugent scoring system. METHODS: This was a descriptive cross-sectional study conducted at Tribhuvan University Teaching Hospital, Nepal from October 2016 to September 2017. Vaginal specimens were collected from a total of 141 women presenting with abnormal vaginal discharge. The sensitivity, specificity, positive predictive value, and negative predictive value of Amsel's criteria were calculated, and each component of Amsel's criteria was compared to the Nugent scoring system. RESULTS: The sensitivity, specificity, positive predictive value, and negative predictive value of Amsel's criteria were 50%, 98.2%, 87.5%, and 88.8% respectively. The clue cells showed 100% specificity and vaginal discharge with pH > 4.5 had 89.3% sensitivity while compared with Nugent's scoring system. CONCLUSIONS: Amsel's criteria can be used as an adjunct method to Nugent scoring for the diagnosis of BV in the hands of skilled manpower in resources limited countries. The presence of clue cell and positive whiff test of Amsel's criteria shows good match with Nugent's score.
Subject(s)
Vaginosis, Bacterial , Adult , Cross-Sectional Studies , Female , Humans , Nepal , Sensitivity and Specificity , Tertiary Care Centers , Vaginosis, Bacterial/diagnosisABSTRACT
Increased airway smooth muscle (ASM) cell mass and secretory functions are characteristics of airway inflammatory diseases, such as asthma. To date, there are no effective therapies to combat ASM cell proliferation, which contributes to bronchoconstriction and airway obstruction. Growth factors such as platelet-derived growth factor (PDGF) and the activation of the ERK1/2 are major regulators of ASM cell proliferation and airway remodeling in asthma. However, given the ubiquitous expression and multiple functions of ERK1/2, complete inhibition of ERK1/2 using ATP-competitive inhibitors may lead to unwanted off-target effects. Alternatively, we have identified compounds that are designed to target substrate docking sites and act as function-selective inhibitors of ERK1/2 signaling. Here, we show that both function-selective and ATP-competitive ERK1/2 inhibitors are effective at inhibiting PDGF-mediated proliferation, collagen production, and IL-6 secretion in ASM cells. Proteomic analysis revealed that both types of inhibitors had similar effects on reducing proteins related to TGF-ß and IL-6 signaling that are relevant to airway remodeling. However, function-selective ERK1/2 inhibitors caused fewer changes in protein expression compared with ATP-competitive inhibitors. These studies provide a molecular basis for the development of function-selective ERK1/2 inhibitors to mitigate airway remodeling in asthma with defined regulation of ERK1/2 signaling.-Defnet, A. E., Huang, W., Polischak, S., Yadav, S. K., Kane, M. A., Shapiro, P., Deshpande, D. A. Effects of ATP-competitive and function-selective ERK inhibitors on airway smooth muscle cell proliferation.
Subject(s)
Bronchi/cytology , Bronchi/metabolism , MAP Kinase Signaling System , Myocytes, Smooth Muscle/cytology , Myocytes, Smooth Muscle/metabolism , Adenosine Triphosphate/metabolism , Airway Remodeling/drug effects , Asthma/metabolism , Asthma/pathology , Cell Proliferation/drug effects , Cells, Cultured , Gene Expression/drug effects , Humans , MAP Kinase Signaling System/drug effects , Myocytes, Smooth Muscle/drug effects , Platelet-Derived Growth Factor/pharmacology , Protein Kinase Inhibitors/pharmacology , Transcription Factor AP-1/metabolismABSTRACT
The transient contamination of medical professional's attires including white coats is one of the major vehicles for the horizontal transmission of microorganisms in the hospital environment. This study was carried out to determine the degree of contamination by bacterial agents on the white coats in a tertiary care hospital in Nepal. Sterilized uniforms with fabric patches of 10 cm × 15 cm size attached to the right and left pockets were distributed to 12 nurses of six different wards of a teaching hospital at the beginning of their work shift. Worn coats were collected at the end of the shifts and the patches were subjected for total bacterial count and identification of selected bacterial pathogens, as prioritized by the World Health Organization (WHO). Fifty percent of the sampled swatches were found to be contaminated by pathogenic bacteria. The average colony growth per square inch of the patch was 524 and 857 during first and second workdays, respectively, indicating an increase of 63.6% in colony counts. The pathogens detected on patches were Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Acinetobacter sp. Additional bacteria identified included Bacillus sp., Micrococcus sp., and coagulase-negative staphylococci (CoNS). The nurses working in the maternity department had their white coats highly contaminated with bacteria. On the other hand, the least bacterial contamination was recorded from the nurses of the surgery ward. One S. aureus isolate from the maternity ward was resistant to methicillin. This study showed that pathogens belonging to the WHO list of critical priority and high priority have been isolated from white coats of nurses, thus posing the risk of transmission to patients. White coats must be worn, maintained, and washed properly to reduce bacterial contamination load and to prevent cross-contamination of potential superbugs. The practice of wearing white coats outside the healthcare zone should be strictly discouraged.
ABSTRACT
Meiosis is the defining event of spermatogenesis. Spermatocytes undergo meiosis to give rise to round spermatids, which in turn metamorphose to flagellated spermatozoa that mature in the epididymis. To characterize the dynamics of gene expression during these important stages of spermatogenesis, we undertook transcriptome analysis in >90% pure pachytene spermatocytes and round spermatids, and pure mature sperm of rat by massive parallel deep sequencing. The study has identified 10,719 total transcripts expressed in meiotic and postmeiotic cells, out of which 7,641 were present in all the three cell types. Most abundant transcripts were related to gametogenesis in spermatocytes and spermatids, and mitochondrial energy metabolism in sperm. Importantly, 108 transcripts were specific to spermatocytes, including Cpeb2, Dpf3, H2afy, Haus7, Plcb1, Taf9, and Tdrd7 strongly linked with meiosis. Similarly, 323 transcripts unique to round spermatids included Arpc5, Apoa1, Cntrob, Dcaf17, Ift88, and Ly6k that play essential roles in spermiogenesis. Likewise, 178 transcripts unique to sperm included Camta1, Hoxb1, and Prdx6 having assigned roles in fertility and/or embryonic development. Levels of ~16% transcripts declined from spermatocytes to sperm while two (Cd300e and Ddx17) increased. New candidate genes with possible roles in meiosis (91), spermiogenesis (298), and sperm function (171), have been identified. This study has provided new potential targets for contraception and/or treatment of male infertility. (CDRI communication number 9889).
Subject(s)
Gene Expression Regulation/physiology , Meiosis/physiology , Spermatocytes/metabolism , Spermatogenesis/physiology , Animals , Male , Rats , Rats, Sprague-Dawley , Spermatocytes/cytologyABSTRACT
MicroRNA (miRNA)-guided mRNA repression, mediated by the miRNA-induced silencing complex (miRISC), is an important component of post-transcriptional gene silencing. However, how miRISC identifies the target mRNA in vivo is not well understood. Here, we show that the nucleoporin Nup358 plays an important role in this process. Nup358 localizes to the nuclear pore complex and to the cytoplasmic annulate lamellae (AL), and these structures dynamically associate with two mRNP granules: processing bodies (P bodies) and stress granules (SGs). Nup358 depletion disrupts P bodies and concomitantly impairs the miRNA pathway. Furthermore, Nup358 interacts with AGO and GW182 proteins and promotes the association of target mRNA with miRISC A well-characterized SUMO-interacting motif (SIM) in Nup358 is sufficient for Nup358 to directly bind to AGO proteins. Moreover, AGO and PIWI proteins interact with SIMs derived from other SUMO-binding proteins. Our study indicates that Nup358-AGO interaction is important for miRNA-mediated gene silencing and identifies SIM as a new interacting motif for the AGO family of proteins. The findings also support a model wherein the coupling of miRISC with the target mRNA could occur at AL, specialized domains within the ER, and at the nuclear envelope.
Subject(s)
Argonaute Proteins/metabolism , MicroRNAs/genetics , Molecular Chaperones/metabolism , Nuclear Pore Complex Proteins/metabolism , Protein Interaction Domains and Motifs , RNA, Messenger/genetics , RNA-Induced Silencing Complex/metabolism , Amino Acid Sequence , Animals , Argonaute Proteins/chemistry , Cell Line , Gene Silencing , Humans , Intranuclear Inclusion Bodies/metabolism , MicroRNAs/metabolism , Models, Biological , Models, Molecular , Molecular Chaperones/chemistry , Nuclear Pore Complex Proteins/chemistry , Protein Binding , Protein Conformation , RNA Interference , RNA, Messenger/metabolism , Signal Transduction , Zinc FingersABSTRACT
AIM: The study was carried to find a relationship between the postextraction stable landmark, the incisive papilla, and the most labial position of the maxillary central incisor teeth, which occurred in Nepalese population. MATERIALS AND METHODS: Casts of the subjects selected by nonprobability random sampling meeting the inclusion criteria were obtained. Each casts were standardized with respect to the occlusal plane and a photographic technique was used to measure the distance from the tangent of the labial surface of the central incisors to the posterior border of the incisive papilla. The measurements were made using Adobe Photoshop and results were analyzed by using appropriate statistical methods. Most appropriate software (SPSS) for the purpose was used to generate all desired values. RESULTS: The data obtained suggested that the distance from the labial surface of maxillary central incisors to the posterior border of the incisive papilla ranged from 9 to 15.9 mm with a mean of 11.59 mm (SD 1.3). Various other results were also found after evaluation of the arch forms in relation to sex and race. CONCLUSION: Within the limitations of the study, these results suggested that there is a relationship between the maxillary central incisors and the incisive papilla aiding in the anteroposterior positioning of the anterior tooth. The clinical relevance of the study lies in application of the incisive papilla as a starting point in the preliminary location of maxillary incisors and canines during construction of the denture in absence of preextraction records.
Subject(s)
Incisor/anatomy & histology , Palate/anatomy & histology , Adolescent , Adult , Asian People , Female , Humans , Male , Maxilla , Nepal , Young AdultABSTRACT
BACKGROUND: The intestinal colonization and transmission of antibiotic-resistant Enterobacteriales to renal transplant recipients may pose a threat to them because they are profoundly immunocompromised and vulnerable to infection. Hence, it is crucial to identify these antibiotic-resistant fecal Enterobacteriales harboring high-risk populations. The objective of this study was to determine antibiotic resistance as well as ß-lactamases production in fecal Enterobacteriales among renal transplant recipients. METHODS: The stool samples, one collected from each transplant recipient, were processed for isolation and identification of Enterobacteriales and were tested for their antibiotic susceptibility, extended-spectrum ß-lactamase, and metallo-ß-lactamase production by standard methods. RESULTS: A total of 103 Enterobacteriales comprising of Escherichia coli (86.4%), Klebsiella species (11.7%), and Citrobacter species (1.9%) were isolated and more than 60% of the E. coli were found resistant to ceftazidime and ciprofloxacin and around half of the Klebsiella species were resistant to ceftazidime and fluroquinolones. The extended-spectrum ß-lactamase production was seen in 3.4% and 8.3% and metallo-ß-lactamase production in 24.7% and 33.3% of E. coli and Klebsiella species, respectively. The high proportion of ß-lactamase-producers were resistant to piperacillin-tazobactam, meropenem, gentamicin, and amikacin than ß-lactamases non-producers. CONCLUSION: Since the antibiotic resistance is higher in fecal Enterobacteriales, each renal transplant recipient should be screened for these highly resistant intestinal colonizers after transplantation in order to prevent infections and to reduce the rate of transplant failure due to infections.
Subject(s)
Anti-Bacterial Agents , Kidney Transplantation , Humans , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/therapeutic use , Ceftazidime , Transplant Recipients , Escherichia coli , Nepal , beta-Lactamases , KlebsiellaABSTRACT
An efficient bacterial strain capable of simultaneous production of lipase and protease in a single production medium was isolated. Thirty six bacterial strains, isolated from diverse habitats, were screened for their lipolytic and proteolytic activity. Of these, only one bacterial strain was found to be lipase and protease producer. The 16S rDNA sequencing and phylogenetic analyses revealed that strain (NSD-09) was in close identity to Pseudomonas aeruginosa. The maximum lipase (221.4 U/ml) and protease (187.9 U/ml) activities were obtained after 28 and 24 h of incubation, respectively at pH 9.0 and 37 °C. Castor oil and wheat bran were found to be the best substrate for lipase and protease production, respectively. The strain also exhibited high tolerance to lead (1450 µg/ml) and chromium (1000 µg/ml) in agar plates. It also showed tolerance to other heavy metals, such as Co(+2) , Zn(+2) , Hg(+2) , Ni(+2) and Cd(+2) . Therefore, this strain has scope for tailing bioremediation. Presumably, this is the first attempt on P. aeruginosa to explore its potential for both industrial and environmental applications.
Subject(s)
Anti-Bacterial Agents/toxicity , Bacterial Proteins/biosynthesis , Endopeptidases/biosynthesis , Lipase/biosynthesis , Metals, Heavy/toxicity , Pseudomonas aeruginosa/drug effects , Pseudomonas aeruginosa/enzymology , Bacterial Proteins/chemistry , Castor Oil/metabolism , Cluster Analysis , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Dietary Fiber/metabolism , Drug Tolerance , Endopeptidases/chemistry , Enzyme Stability , Hydrogen-Ion Concentration , Lipase/chemistry , Molecular Sequence Data , Phylogeny , Pseudomonas aeruginosa/classification , Pseudomonas aeruginosa/isolation & purification , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , TemperatureABSTRACT
Alkaline lipase production by mutant strain of Pseudomonas aeruginosa MTCC 10,055 was optimized in shake flask batch fermentation using response surface methodology. An empirical model was developed through Box-Behnken experimental design to describe the relationship among tested variables (pH, temperature, castor oil, starch and triton-X-100). The second-order quadratic model determined the optimum conditions as castor oil, 1.77 mL.L(-1); starch, 15.0 g.L(-1); triton-X-100, 0.93 mL.L(-1); incubation temperature, 34.12 °C and pH 8.1 resulting into maximum alkaline lipase production (3142.57 U.mL(-1)). The quadratic model was in satisfactory adjustment with the experimental data as evidenced by a high coefficient of determination (R(2)) value (0.9987). The RSM facilitated the analysis and interpretation of experimental data to ascertain the optimum conditions of the variables for the process and recognized the contribution of individual variables to assess the response under optimal conditions. Hence Box-Behnken approach could fruitfully be applied for process optimization.
ABSTRACT
Organic compound-based nonlinear optical (NLO) materials have sparked a lot of attention due to their multitude of applications and shorter optical response times than those of inorganic NLO materials. In the present investigation, we designed exo-exo-tetracyclo[6.2.1.13,6.02,7]dodecane (TCD) derivatives, which were obtained by replacing H atoms of methylene bridge carbon with alkali metals (Li, Na, and K). It was observed that upon the substitution of alkali metals at bridging CH2 carbon, absorption within the visible region occurred. Moving from 1 to 7 derivatives, the maximum absorption wavelength of the complexes exhibited a red shift. The designed molecules showed a high degree of intramolecular charge transfer (ICT) and excess electrons in nature, which were responsible for rapid optical response time and significant large molecular (hyper)polarizability. Calculated trends also inferred that the crucial transition energy decreased in order that also played a key role in the higher nonlinear optical response. Furthermore, to examine the effect of the structure/property relationship on the nonlinear optical properties of these investigated compounds (1-7), we calculated the density of state (DOS), transition density matrix (TDM), and frontier molecular orbitals (FMOs). The largest first static hyperpolarizability (ßtot) of TCD derivative 7 was 72059 au, which was 43 times greater than that of the prototype p-nitroaniline (ßtot = 1675 au).
ABSTRACT
Human activity recognition (HAR) using drone-mounted cameras has attracted considerable interest from the computer vision research community in recent years. A robust and efficient HAR system has a pivotal role in fields like video surveillance, crowd behavior analysis, sports analysis, and human-computer interaction. What makes it challenging are the complex poses, understanding different viewpoints, and the environmental scenarios where the action is taking place. To address such complexities, in this paper, we propose a novel Sparse Weighted Temporal Attention (SWTA) module to utilize sparsely sampled video frames for obtaining global weighted temporal attention. The proposed SWTA is comprised of two parts. First, temporal segment network that sparsely samples a given set of frames. Second, weighted temporal attention, which incorporates a fusion of attention maps derived from optical flow, with raw RGB images. This is followed by a basenet network, which comprises a convolutional neural network (CNN) module along with fully connected layers that provide us with activity recognition. The SWTA network can be used as a plug-in module to the existing deep CNN architectures, for optimizing them to learn temporal information by eliminating the need for a separate temporal stream. It has been evaluated on three publicly available benchmark datasets, namely Okutama, MOD20, and Drone-Action. The proposed model has received an accuracy of 72.76%, 92.56%, and 78.86% on the respective datasets thereby surpassing the previous state-of-the-art performances by a margin of 25.26%, 18.56%, and 2.94%, respectively.
Subject(s)
Benchmarking , Unmanned Aerial Devices , Humans , Learning , Neural Networks, Computer , Recognition, PsychologyABSTRACT
Antimicrobial resistance (AMR) is increasing and represents one of the greatest public health challenges of our time, accounting for considerable morbidity and mortality globally. A "One Health" surveillance strategy, which integrates data concerning the resistant organisms circulating in humans, animals, and the environment, is required to monitor this issue and enable effective interventions. The timely collection, processing, analysis, and reporting of AMR surveillance data are necessary for the effective delivery of the information generated from such surveillance. Nepal has greatly improved its surveillance activities through a network of human and animal health laboratories; however, the data reported by sentinel laboratories are often inconsistent, incomplete, and delayed, causing challenges in terms of data cleaning, standardization, and visualization on a national level. To overcome these issues, innovative methods and procedures have been adopted in Nepal, with the development and customization of digital tools that reduce the human time and effort spent on data cleaning and standardization, with concomitant improvements in the accuracy of data. These standardized data can be uploaded to the district health information system 2 (DHIS2) One Health AMR surveillance portal, enabling the generation of reports that will help decision-makers and policy planners to combat the global problem of AMR.
ABSTRACT
A suitable substitution of carbazole with a π-spacer group like cyanoethynylethene offers exciting future opportunities in terms of smart nonlinear optical material. In the quest of better organic nonlinear optical material, we have designed a series of derivatives based on carbazole and cyanoethynylethene fragment combinations in a unique fashion by employing the density functional (DFT) methods. The calculated time-dependent density functional theory (TD-DFT) calculations infer that the gigantic first static hyperpolarizability (ßtot) values are due to a lower energy gap and higher transition dipole moment for the crucial electronic transition. Furthermore, to see the in-depth execution for enhanced second-order nonlinear optics and the structure property relationship on nonlinear optics (NLO) behavior, we have performed frontier molecular orbitals (FMO), density of state (DOS), and transition density matrix (TDM). Furthermore, CAM-B3LYP functional-based calculated results infer that the designed molecule 10 show the first static hyperpolarizability is 923.93 × 10-30 esu which is 69 times larger than that of p-nitroaniline.
ABSTRACT
Background and Objectives: In recent decades, the incidence of dengue has increased dramatically. In dengue-endemic countries, changes in dengue virus serotypes, genotypes, and lineages have been reported. This study was designed to detect and characterize the dengue virus isolates circulating in North India by serological and molecular techniques. Materials and Methods: This study was conducted at the Post Graduate Institute of Medical Education and Research (PGIMER), Chandigarh, India. NS1 antigen and IgM antibody against dengue were detected by ELISA methods, viral RNA was extracted and amplified by conventional PCR and one-step single-tube multiplex PCR. The purified PCR products were cycle sequenced and a database search was implemented for the confirmation of the sequence product. Phylogenetic analysis was carried out with previously reported sequences. Results: Among 1509 samples, 205 (13.6%) were found positive for IgM antibodies with the highest number (n=67) among the 21 to 30 years age group with peak positivity during post-monsoon months. Among acute samples, NS1 antigen was positive in 62.9%. Seven patients out of 13 had dengue viral RNA in PCR. It comprised six DENV-2 serotypes and one DENV-3 serotype. On phylogenetic analysis, DENV-2 strains grouped with genotype IV and DENV-3 with genotype III. Conclusion: Dengue infection was found frequently during post-monsoon season. The positivity rate of the dengue NS1 antigen test was greater than that of the antibody test. The dengue isolates were characterized as genotype IV and genotype III of DENV-2 and DENV-3 respectively.