Effects of high glutamate concentrations on mitochondria of human neuroblastoma SH-SY5Y cells.

BACKGROUND: The excess amount of glutamate in neurons is associated with the excitotoxicity and neurodegenerative diseases. Glutamate induces neurotoxicity primarily by immense influx of Ca2+ arising from overstimulation of the NMDA subtype of glutamate receptors. The neuronal death induced by the overstimulation of glutamate receptors depends critically on a sustained increase in mitochondrial Ca2+ influx and impairment in mitochondrial functions. The mitochondrial impairment is an important contributor to the glutamate-induced neuronal toxicity and thus provides an important target for the intervention. The present study investigates the effects of high glutamate concentrations on mitochondrial functions. RESULTS: Here, we have shown that the higher concentration of glutamate treatment caused a significant elevation in the N-methyl-D-aspartate (NMDA) receptors expression and elevated the intra-mitochondrial calcium accumulation in SHSY5Y neuronal cells. As a result of an accumulation of intra-mitochondrial calcium, there is a concentration-dependent elevation in ROS in the mitochondria. Tyrosine nitration of several mitochondrial proteins was increased while the mitochondrial membrane potential was dissipated. Furthermore, glutamate treatments also resulted in mitochondrial membrane permeability transition. CONCLUSIONS: These findings suggest that treatment of high glutamate concentration causes impairment of mitochondrial functions by an increase in intra-mitochondrial calcium, ROS production, dissipation of mitochondrial membrane potential and mitochondrial permeability transition pore opening in human neuroblastoma SHSY5Y cells.

Vaginal Atresia with Cervical Agenesis, a Rare Congenital Malformation.

Congenital vaginal atresia and cervical agenesis is a rare congenital abnormality of the female genital tract. Here we report a case of 15-year old girl with primary amenorrhea with hematometra, presented with lower abdominal mass. She was symptomatic since 5 months and visited local hospital after 4 months of onset of her symptoms when it became severe, where diagnostic laparotomy was performed for suspected Adnexal mass. Intraoperatively adnexal mass was adhered with and extended up to the uterus with 16 weeks size of uterus. Abdomen was closed without any further intervention and was referred to higher center for needful. When she presented to Kathmandu Model Hospital, she was asymptomatic on her 5th post-operative day of laparotomy. We planned for surgical intervention after examination and investigation. Drainage with vaginoplasty with amnion graft with placement of mould was done.

COVID-19 Pandemic: An Experience of OBGYN Residents at Kathmandu Model Hospital.

COVID-19, a novel corona virus has affected the life of each and every individual worldwide. Nepal being the neighborhood country of china, though, we had a late case detection. But, since the month of July this virus has spread in an alarming manner in Nepal. Nepal being one of the developing countries, we lack in equipments, manpower resources and also in treatment centers. Looking into the devastating scenario of Covid 19 in China, Italy, New York, Brazil and our neighboring countries like India, Pakistan and Bangladesh is scary. We wondered how we were going to handle this pandemic if similar circumstances happened in our country too. At the same time being OBGYN residents, we all know pregnancy is very crucial and our patients have faced much difficulties to receive the maternal health services. In this manuscript, we have shared our experience regarding preparedness for COVID-19, management of positive cases and its effect in OBGYN trainee.

Misleading Diagnosis of Dysgerminoma in a Young Asymptomatic Patient.

Dysgerminomas account for approximately one third of all malignant ovarian germ cell tumors (tumors arising from ovarian germinal elements) and are the most common ovarian malignancy detected during pregnancy. They are the only germ cell malignancy with a significant rate of bilateral ovarian involvement that is 15-20 percent. They have a variable gross appearance, but in general are solid, pink to tan to cream colored lobulated masses. They have the best prognosis of all malignant ovarian germ cell tumor variants. Two thirds are stage I at diagnosis, and prognosis is excellent even for those with advanced disease due to exquisite tumor chemosensitivity. The 5 year disease specific survival rate approximates 99 percent. This is a case report of a huge ovarian dysgerminoma in a young unmarried lady that was quite asymptomatic. She underwent laparotomy with right ovarian cystectomy.

Pregnancy Complicated by Acute Pancreatitis with Ascariasis.

Ascariasis lumbricoides infections are common in tropical and sub-tropical countries. As it is one of the common causes for common bile duct obstruction. But in pregnancy is a rare entity. It may present with complications like acute pancreatitis. Here we present a case of 24 year female gravida 2 para 1 at 34 weeks of pregnancy presented with chief complain of pain over epigastric region radiating to back and associated with two episodes of vomiting, non bilious, not mixed with blood. Ultrasonography showed long tubular hyperechoic structure in gallbladder lumen most likely ascariasis and then she was diagnosed as a case of acute pancreatitis with alive ascariasis.

Predicting non-point source of pollution in Maithon reservoir using a semi-distributed hydrological model.

Non-point source (NPS) pollution has been emerged as a major cause for reduced water quality of a lake due to increased human interference and disturbances in the natural condition of the surrounding catchment. The impact is, even more, worsening in the monsoon season when there is increased surface runoff. In the present study, an attempt has been made to predict the seasonal (monsoon) NPS loading in terms of sediment, nitrogen, and phosphorous in Maithon reservoir using Soil and Water Assessment Tool (SWAT) hydrologic model. The SWAT model was initially calibrated using monthly runoff and sediment yield data of monsoon period for the year 1998-2005 using observed data of Rajdhanwar station followed by its validation for the observed monthly runoff and sediment data from Giridih and Santrabad for the same duration. The calibrated SWAT model was used to predict the sediment, total nitrogen, and phosphorous influx in the Maithon reservoir. It has been observed that average sediment yield from different micro-watersheds varies from 0.231 to 7.458 ton/ha, while average monthly nitrogen and phosphorous yields vary from 0.224 to 1.377 kg/ha and 0.073 to 0.363 kg/ha, respectively, during the monsoon period. On the other hand, the net monthly average sediment yield and total nitrogen and phosphorous yields in the reservoir were found to be 1.53 M ton, 1834.2 kg, 191.1 kg, respectively. The results indicate there is a substantial influx of nutrients and sediments into the Maithon reservoir. The study not only provides insights on the potential NPS pollutant loading in the reservoir but also enables to identify the hotspot of NPS pollution where immediate mitigation measures have to be taken at priority basis.

Lead Tolerance and its Accumulation by a Tree Legume: Dalbergia sissoo DC.

Dalbergia sissoo DC, a leguminous tropical timber tree has been investigated against the Pb toxicity; under the Pb-stress, plant's morphology, biochemical parameters and genomic template stability (GTS) screened in vitro. At the optimum Pb tolerance level (150 mg L-1), plant's defense mechanism-superoxide dismutase, catalase, ascorbate peroxidases and proline could trigger to achieve optimum vegetative growth with minimum fluctuations of the GTS. Further, D. sissoo roots could accumulate 2399.8 ± 16 mg kg-1 Pb. Scanning electron microscopy and energy dispersive X-ray spectrometer analysis also revealed the deposition of Pb in root tissues. In a 1 year pot experiment with Pb-contaminated soil, the plants exhibited normal growth, and Pb accumulation significantly enhanced by the amalgamation of citric acid in the soil. Thus, the tree may prove as a potential candidate for Pb phytostabilization.

Analysis of A549 cell proteome alteration in response to recombinant influenza A virus nucleoprotein and its interaction with cellular proteins, a preliminary study.

Influenza A virus undergoes frequent changes of antigenicity and contributes to seasonal epidemics or unpredictable pandemics. Nucleoprotein, encoded by gene segment 5, is an internal protein of the virus and is conserved among strains of different host origins. In the current study, we analyzed the differentially expressed proteins in A549 cells transiently transfected with the recombinant nucleoprotein of influenza A virus by 2D gel electrophoresis. The resolved protein spots on gel were identified by MALDI-TOF/Mass spectrometry analysis. The majority of the host proteins detected to be differentially abundant in recombinant nucleoprotein-expressing cells as compared to vector-transfected cells are the proteins of metabolic pathways, glycolytic enzymes, molecular chaperones and cytoskeletal proteins. We further demonstrated the interaction of virus nucleoprotein with some of the identified host cellular proteins. In vitro binding assay carried out using the purified recombinant nucleoprotein (pET29a+NP-His) and A549 cell lysate confirmed the interaction between nucleoprotein and host proteins, such as alpha enolase 1, pyruvate kinase and ß-actin. The preliminary data of our study provides the information on virus nucleoprotein interaction with proteins involved in glycolysis. However, studies are ongoing to understand the significance of these interactions in modulating the host factors during virus replication.

Non-Surgical Pneumoperitoneum.

Pneumoperitoneum is mostly caused by visceral perforation and surgical intervention; however non-surgical pneumoperitoneum has been reported without evidence of visceral disease. Blunt chest trauma causing an abrupt rise in thoracic pressure can leak air through the microscopic diaphragmatic defects or the mediastinum along perivascular connective tissue and cause pneumoperitoneum. We hereby present a case of non-surgical pneumoperitoneum after blunt chest trauma that was brought to the emergency department of college of medical sciences teaching hospital with features of bilateral pneumothorax with subcutaneous emphysema and abdominal distension which was diagnosed and managed promptly with bilateral chest drain and other supportive treatments.

In vitro activity of bioactive extracts from rare actinomycetes against multi-drug resistant Streptococcus pneumoniae.

AIMS: In this study, we investigated the in vitro potential of the bioactive extracts from five putatively novel species of actinomycetes isolated from the Indian hot desert against multi-drug resistant (MDR) Streptococcus pneumoniae. METHODS AND RESULTS: The antimicrobial activity of 10 different extracts was evaluated against S. pneumoniae strains with, erm(B) and mef(E) genes as well as fluoroquinolone-resistant (FQ(R) ) strains using the micro-broth dilution method. Of these 10 extracts, four exhibited good to excellent anti-S. pneumoniae activity with minimum inhibitory concentrations (MICs) ranging from 0·125 to 8 µg ml(-1) . The time-kill kinetics study showed that these extracts killed the pathogens in 2-8 h. In vitro cell-free transcription/translation of luciferase gene using S30 bacterial extract and TNT mammalian ribosome indicated that they inhibited bacterial ribosomes at much lower concentrations than those required to inhibit the mammalian ribosomes. CONCLUSIONS: This study demonstrates that these are potent concentration-dependent bactericidal metabolites with 16-fold higher in vitro activity than levofloxacin against MDR S. pneumoniae. SIGNIFICANCE AND IMPACT OF THE STUDY: Metabolites from actinomycetes can be excellent inhibitors of MDR S. pneumoniae. Considering the in vitro efficacy of these crude extracts against S. pneumoniae MDR spp., once purified these can be used against streptococcal pathogens causing community-acquired pneumonia.

Production of Biohydrogen from Wastewater by Klebsiella oxytoca ATCC 13182.

Production of biohydrogen from distillery effluent was carried out by using Klebsiella oxytoca ATCC 13182. The work focuses on optimization of pH, temperature, and state of bacteria, which are the various affecting factors for fermentative biohydrogen production. Results indicates that at 35 °C for suspended cultures, the production was at its maximum (i.e., 91.33 ± 0.88 mL) when compared with other temperatures. At 35 °C and at pH 5 and 6, maximum productions of 117.67 ± 1.45 and 111.67 ± 2.72 mL were observed with no significant difference. When immobilized, Klebsiella oxytoca ATCC 13182 was used for biohydrogen production at optimized conditions, production was 186.33 ± 3.17 mL. Hence, immobilized cells were found to be more advantageous for biological hydrogen production over suspended form. Physicochemical analysis of the effluent was conducted before and after fermentation and the values suggested that the fermentative process is an efficient method for biological treatment of wastewater.

Face-specific identification impairments following sight-providing treatment may be alleviated by an initial period of low visual acuity.

Identifying faces requires configural processing of visual information. We previously proposed that the poor visual acuity experienced by newborns in their first year of life lays the groundwork for such configural processing by forcing integration over larger spatial fields. This hypothesis predicts that children treated for congenital cataracts late in life will exhibit persistent impairments in face- but not object-identification, because they begin their visual journey with higher than newborn acuity. This would not be the case for patients whose pretreatment condition has allowed for initial low acuity vision, like that of a newborn. Here, we test this prediction by assessing the development of facial identification skill in three groups: patients treated for congenital cataracts whose pretreatment visual acuity was worse than that of a newborn, patients whose pretreatment acuity was better than that of a newborn, and age-matched controls. We find that while both patient groups show significant gains in object-identification, the emergence of face identification is determined by pretreatment acuity: patients with pre-operative acuity worse than a newborn did not show any improvements on face-identification tasks despite years of visual experience, whereas those with pretreatment acuity comparable to a newborn improved on both the object- and face-identification tasks. These findings not only answer our research question but also provide new insights into the role of early visual acuity in facial identification development. We discuss these results in the context of both typical and atypical visual development.

Chromium phytotoxicity in radish (Raphanus sativus): effects on metabolism and nutrient uptake.

In the present investigation, chromium (VI) induced toxicity on metabolic activity and translocations of nutrients in radish were evaluated under controlled glass house conditions. Chromium was found to induce toxicity and significantly affect plant growth and metabolic activity. Excess of chromium (0.4 mM) caused a decrease in the concentration of iron in leaves (from 134.3 to 71.9 µg g(-1) dw) and significant translocation of sulphur, phosphorus and zinc. Translocation of manganese, copper and boron were less affected from root to stem. After 15 days of Cr exposure, maximum accumulation of Cr was found in roots (327.6 µg g(-1) dw) followed by stems (186.8 µg g(-1) dw) and leaves (116.7 µg g(-1) dw) at 0.4 mM Cr concentration. Therefore, Cr may affect negatively not only production, but also the nutritive quality of the radish; likewise, higher Cr content may cause health hazards for humans.

Human fascioliasis with biliary complications.

We report a case of human fascioliasis adding to the few of the previously reported cases in India. A young boy from rural background in Bihar presented with diarrhea, vomiting, hepatic tenderness, jaundice and fever along with peripheral eosinophilia. Examination of stool revealed yellow-brown eggs of Fasciola hepatica. Human fascioliasis should be kept in mind in patients with cholangitis and eosinophilia especially in areas of sporadic occurrence.

Glutaredoxin proteins from E. coli isoforms were compared in terms of energy frustration.

Glutaredoxin (GRXs) protein plays a vital role inside the cell, including redox control of transcription to the cell's antioxidant defense, apoptosis, and cellular differentiation regulation. In this study, we have investigated the energy landscape and characterized the pattern of local frustration in different forms and states of the GRX protein ofE. coli.Analysis was done on the conformational alterations, significant changes in the frustration pattern, and different GRXs such as GRX-II, GRX-III, GRX-II-GSH, and GRX-III-GSH complex. We have found the practice of frustration, and structure was quite similar in the same isoform having different states of protein; however, a significant difference was observed between different isoforms. Moreover, oxidation of GRX-I introduced an extra α-helix increasing the destabilizing interactions within the protein. The study of frustrated contacts on oxidized and reduced GRX and with bound and unbound Glutathione indicates its potential application in activating and regulating the behavior of GRXs.

Development and characterization of microsatellite markers from tropical forage Stylosanthes species and analysis of genetic variability and cross-species transferability.

A limited number of functional molecular markers has slowed the desired genetic improvement of Stylosanthes species. Hence, in an attempt to develop simple sequence repeat (SSR) markers, genomic libraries from Stylosanthes seabrana B.L. Maass & 't Mannetje (2n=2x=20) using 5' anchored degenerate microsatellite primers were constructed. Of the 76 new microsatellites, 21 functional primer pairs were designed. Because of the small number of primer pairs designed, 428 expressed sequence tag (EST) sequences from seven Stylosanthes species were also examined for SSR detection. Approximately 10% of sequences delivered functional primer pairs, and after redundancy elimination, 57 microsatellite repeats were selected. Tetranucleotides followed by trinucleotides were the major repeated sequences in Stylosanthes ESTs. In total, a robust set of 21 genomic-SSR (gSSR) and 20 EST-SSR (eSSR) markers were developed. These markers were analyzed for intraspecific diversity within 20 S. seabrana accessions and for their cross-species transferability. Mean expected (He) and observed (Ho) heterozygosity values with gSSR markers were 0.64 and 0.372, respectively, whereas with eSSR markers these were 0.297 and 0.214, respectively. Dendrograms having moderate bootstrap value (23%-94%) were able to distinguish all accessions of S. seabrana with gSSR markers, whereas eSSR markers showed 100% similarities between few accessions. The set of 21 gSSRs, from S. seabrana, and 20 eSSRs, from selected Stylosanthes species, with their high cross-species transferability (45% with gSSRs, 86% with eSSRs) will facilitate genetic improvement of Stylosanthes species globally.

Metal contamination of soil and translocation in vegetables growing under industrial wastewater irrigated agricultural field of Vadodara, Gujarat, India.

The present investigation was carried out to evaluate metals concentration in ten vegetable crops growing in mixed industrial effluent irrigated agricultural field near Vadodara, Gujarat, India. Differential accumulation and translocation of various metals in selected vegetables plant species was observed. A higher concentration of metals were found in order of Fe>Mn>Zn>Cd>Cu>Pb>Cr>As in soil irrigated with industrial effluent than soil irrigated with tube well water; however, the concentration of As, Cr and Pb found below detection limit in tube well water irrigated soil. Metal accumulation in root and top of vegetables varied significantly both in relations to metal concentration in the soil and the plant genotype. Among ten vegetable species studied five vegetable species, i.e. Spinach, Radish, Tomato, Chili and Cabbage growing in mixed industrial effluent irrigated agricultural field showed high accumulation and translocation of toxic metals (As, Cd, Cr, Pb and Ni) in their edible parts, thus, their cultivation are unsafe with respect to possible transfer in food chain and health hazards. However, it is suggested that vegetable crops restricting toxic metal in non-edible port may be recommended for cultivation in such metal contaminated agricultural field.

Anthropogenic nitrate in groundwater and its health risks in the view of background concentration in a semi arid area of Rajasthan, India.

An increased nitrate (NO3-) concentration in groundwater has been a rising issue on a global scale in recent years. Different consumption mechanisms clearly illustrate the adverse effects on human health. The goal of this present study is to assess the natural and anthropogenic NO3- concentrations in groundwater in a semi arid area of Rajasthan and its related risks to human health in the different groups of ages such as children, males, and females. We have found that most of the samples (n = 90) were influenced by anthropogenic activities. The background level of NO3- had been estimated as 7.2 mg/L using a probabilistic approach. About 93% of nitrate samples exceeded the background limit, while 28% of the samples were beyond the permissible limit of 45 mg/L as per the BIS limits. The results show that the oral exposure of nitrate was very high as compare to dermal contact. With regard to the non-carcinogenic health risk, the total Hazard Index (HITotal) values of groundwater nitrate were an average of 0.895 for males, 1.058 for females, and 1.214 for children. The nitrate health risk assessment shows that about 38%, 46%, and 49% of the samples constitute the non-carcinogenic health risk to males, females, and children, respectively. Children were found to be more prone to health risks due to the potential exposure to groundwater nitrate.

Biodegradation of phenol by native microorganisms isolated from coke processing wastewater.

The present investigation was undertaken to assess the biodegradation of phenol by native bacteria strains isolated from coke oven processing wastewater. The strains were designated ESDSPB1, ESDSPB2 and ESDSPB3 and examined for colony morphology Gram stain characters and biochemical tests. Phenol degrading performance of all the strains was evaluated initially. One of the strains namely ESDSPB2 was found to be highly effective for the removal of phenol, which was used as sole carbon and energy source. From an initial concentration of 200 mg I(-1) it degraded to 79.84 +/- 1.23 mg l(-1). In turn the effect of temperature (20 to 45 degrees C), pH (5-10) and glucose concentration (0, 0.25 and 0.5%) on the rate of phenol degradation by that particular strain was investigated. Observations revealed that the rate of phenol biodegradation was significantly affected by pH, temperature of incubation and glucose concentration. The optimal conditions for phenol removal were found to be pH of 7 (84.63% removal), temperature, 30 degrees C (76.69% removal) and 0.25% supplemented glucose level (97.88% removal). The main significance of the study is the utilization of native bacterial strains from the waste water itself having potential of bioremediation.

Assessment of hydrochemical backgrounds and threshold values of groundwater in a part of desert area, Rajasthan, India.

Natural background levels (NBLs) and threshold values (TVs) are crucial parameters for identification and the quantification of groundwater pollution, and the evaluation of pollution control measures. The cumulative probability distribution technique was used for the evaluation of NBLs for 36 samples collected during two climate conditions in the part of the desert area from Rajasthan, India. The NBLs for Na+, Cl-, SO42-, HCO3-, NO3- and F- ions were assessed and compared with the natural and anthropogenic processes. The TVs were also calculated for Na+, Cl-, SO42-, HCO3-, NO3- and F- ions, and compared with the drinking limits of the Bureau of Indian Standards. Additionally, the pollution percentage (%) at the individual well was estimated and identified the polluted zones. Results indicate that most of the polluted areas were situated in the southern part, which was influenced by the natural and anthropogenic factors. The sodium concentrations above the TVs, in indicating the saline nature of water. Chloride threshold value above the drinking water limit was mainly observed in the dry season, related to intensive evaporation and industrial waste, which leads to groundwater quality degradation. The NO3- concentration (∼56% samples) above the TVs indicates extensive use of nitrate fertilizers and sewage effluent. The values of total dissolved solids (TDS) shows the suspicious scenario as about 84% of the samples in the dry period and about 89% in the wet season exceeding the drinking limit. Assessment of background concentrations and threshold values on regional and local scale assigns the basis for the identification of groundwater pollution, and helpful for better water quality guidelines to protecting of water resources.