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.

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.

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.

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.

Design, characterization and in vitro evaluation of thin films enriched by tannic acid complexed by Fe(III) ions.

Materials based on carbohydrate polymers may be used for biomedical application. However, materials based on natural polymers have weak physicochemical properties. Thereby, there is a challenge to improve their properties without initiation of toxicity. The alternative method compared to toxic chemical agents' addition is the use of metal complexation method. In this study, chitosan/tannic acid mixtures modified by Fe(III) complexation are proposed and tested for potential applications as wound dressings. Thereby, surface properties, blood compatibility as well as platelet adhesion was tested. In addition, the periodontal ligament stromal cells compatibility studies were carried out. The results showed that the iron(III) addition to chitosan/tannic acid mixture improves properties due to a decrease in the surface free energy and exhibited a reduction in the hemolysis rate (below 5%). Moreover, cells cultured on the surface of films with Fe(III) showed higher metabolic activity. The current findings allow for the medical application of the proposed materials as wound dressings.

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.

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.

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.

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.

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.

Prevalence of Cryptosporidium spp. in asymptomatic small ruminants in Grenada, West Indies.

Cryptosporidium spp. is a protozoan parasite that causes enteric infection in a wide range of hosts, including livestock and humans. The aim of this cross-sectional study was to estimate the prevalence of Cryptosporidium spp. in small ruminants in Grenada, West Indies. Fecal samples were collected from 100 sheep and 202 goats from 32 farms. The fecal samples were tested using an Enzyme-linked immunosorbent assay (ELISA) for qualitative detection of antigens in feces (Diagnostic Automation Inc., USA). The overall prevalence of Cryptosporidium spp. was 19.5% [95% confidence interval (CI): 15.4% to 24.4%] in both sheep and goats. The prevalence of Cryptosporidium spp. in sheep and goats was 14% (95% CI: 8.4% to 22.3%) and 22.3% (95% CI: 17.1% to 28.5%), respectively. There was no significant difference in the prevalence of Cryptosporidium spp. infection between sheep and goats (p = .42, Fisher's exact test) in Grenada. Of the 32 farms visited, 19 (59.4%) had at least one Cryptosporidium spp. positive animal.

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.

Evaluation of highly conserved hsp65-specific nested PCR primers for diagnosing Mycobacterium tuberculosis.

OBJECTIVE: To evaluate the sensitivity and specificity of a new nested set of primers designed for the detection of Mycobacterium tuberculosis complex targeting a highly conserved heat shock protein gene (hsp65). DESIGN: The nested primers were designed using multiple sequence alignment assuming the nucleotide sequence of the M. tuberculosis H37Rv hsp65 genome as base. Multidrug-resistant Mycobacterium species along with other non-mycobacterial and fungal species were included to evaluate the specificity of M. tuberculosis hsp65 gene-specific primers. RESULTS: The sensitivity of the primers was determined using serial 10-fold dilutions, and was 100% as shown by the bands in the case of M. tuberculosis complex. None of the other non M. tuberculosis complex bacterial and fungal species yielded any band on nested polymerase chain reaction (PCR). The first round of amplification could amplify 0.3 ng of the template DNA, while nested PCR could detect 0.3 pg. CONCLUSION: The present hsp65-specific primers have been observed to be sensitive, specific and cost-effective, without requiring interpretation of biochemical tests, real-time PCR, sequencing or high-performance liquid chromatography. These primer sets do not have the drawbacks associated with those protocols that target insertion sequence 6110, 16S rDNA, rpoB, recA and MPT 64.

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.

Rhabdomyosarcoma mimicking as pleural effusion.

Rhabdomyosarcoma (RMS), a malignant neoplasm of skeletal muscle origin, is the most common soft tissue sarcoma caused by infectious disease etiology, especially in nondeveloped countries. Despite being a relatively rare cancer, it accounts for approximately 40% of all recorded soft tissue sarcomas. Alveolar RMSs are seen to occur in children, while around 80% cases occur in the first three decades of life. We present here, a case of a 12-year-old child having alveolar RMS, presenting clinically and radiologically with pleural effusion.

Evaluation of Distensibility and Stiffness of Ascending Aortic Aneurysm using Magnetic Resonance Imaging.

INTRODUCTION: Magnetic resonance imaging emerging as a new tool for the diagnosis and evaluation of ascending aortic aneurysm. The aim of our study is to evaluate in vivo distensibility and pulse wave velocity of the aortic wall using functional magnetic resonance imaging technique. METHODS: We enrolled 25 patients undergoing surgery for ascending aortic aneurysm and or aortic valve replacement for a period of 8 months. Preoperatively, all the patients underwent functional MRI study of the aorta. Aortic wall distensibility and pulse wave velocity of ascending aorta was evaluated. RESULTS: Mean age of the patient was 66 years (66.68 ± 5.62 years) with 60% (15) male patients. More than fifty percentages of patients were smoker (52%), hypertensive (64%) and diabetic (56%). We have observed significant decrease of distensibilty in the patients with aortic diameter above 50 mm (p-0.0002). Furthermore, we have found a significant inverse correlation between aortic distensibility and pulse wave velocity (R= -0.650, R2= 0.42, p-0.0004). Similarly, we have found a significant inverse correlation between ascending aortic diameter and distensibility of the aorta (R= -0.785, R2= 0.61, p-0.00001). Statistically significant positive correlation was observed between aortic diameter and pulse wave velocity (R= 0.865, R2= 0.74, p-0.00001). CONCLUSIONS: MRI measurement of aortic diameters, distensibility, and flow wave velocity is an easy, reliable and reproducible technique. Distensibility and pulse wave velocity define the elasticity of the aorta. We have observed that elasticity of aortic wall is decreased in ascending aorta aneurysm patients.

Serological evidence of antibodies to Neospora caninum in stray and owned Grenadian dogs.

Neospora caninum causes abortion in cattle and neuromuscular disease in dogs, world wide. Cattle become infected by ingesting oocysts voided by dogs. The aim of this study was to estimate the seroprevalence of Neospora caninum in two populations of dogs (stray and owned) in Grenada, West Indies. Sera were collected from 625 dogs from all parishes in Grenada. Three hundred and sixty eight dogs were stray, while 257 dogs were owned. Sera were tested for the presence of antibodies against N. caninum using an indirect enzyme linked immunosorbant assay (ELISA) IDvet, France. Antibodies to N. caninum were found in 6 (1.6%) (95% confidence interval (CI): 0.32% to 2.88%) of the stray dogs and in 3 (1.2%, 95% CI: 0.13% to 2.53%) of the owned dogs. Seroprevalence did not differ significantly between the two populations (p=0.74) and between the males and females (p=1). These results suggest that the prevalence of N. caninum infection in dogs in Grenada is low.

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.