Noradrenaline enhances Na-K ATPase subunit expression by HuR-induced mRNA stabilization and their transportation to the cell surface through PLC and PKC mediated pathway: Implications with REMS-loss associated disorders.

Rapid eye movement sleep (REMS) maintains brain excitability at least by regulating Na-K ATPase activity. Although REMS deprivation (REMSD)-associated elevated noradrenaline (NA) increases Na-K ATPase protein expression, its mRNA transcription did not increase. We hypothesized and confirmed both in vivo as well as in vitro that elevated mRNA stability explains the apparent puzzle. The mRNA stability was measured in control and REMSD rat brain with or without in vivo treatment with α1-adrenoceptor (AR) antagonist, prazosin (PRZ). Upon REMSD, Na-K ATPase α1-, and α2-mRNA stability increased significantly, which was prevented by PRZ. To decipher the molecular mechanism of action, we estimated NA-induced Na-K ATPase mRNA stability in Neuro-2a cells under controlled conditions and by transcription blockage using Actinomycin D (Act-D). NA increased Na-K ATPase mRNA stability, which was prevented by PRZ and propranolol (PRP, ß-AR antagonist). The knockdown assay confirmed that the increased mRNA stabilization was induced by elevated cytoplasmic abundance of Human antigen R (HuR) and involving (Phospholipase C) PLC-mediated activation of Protein Kinase C (PKC). Additionally, using cell-impermeable Enz-link sulfo NHS-SS-Biotin, we observed that NA increased Na-K ATPase α1-subunits on the Neuro-2a cell surface. We conclude that REMSD-associated elevated NA, acting on α1- and ß-AR, increases nucleocytoplasmic translocation of HuR and increases Na-K ATPase mRNA stability, resulting in increased Na-K ATPase protein expression. The latter then gets translocated to the neuronal membrane surface involving both PKC and (Protein Kinase A) PKA-mediated pathways. These findings may be exploited for the amelioration of REMSD-associated chronic disorders and symptoms.

Bulbar Dysfunction in Idiopathic Normal Pressure Hydrocephalus: A Case Report.

Normal pressure hydrocephalus (NPH) is a rare condition characterized by pathologically enlarged ventricles and a normal cerebrospinal fluid (CSF) opening pressure measured by lumbar puncture. NPH typically presents as a triad of cognitive decline, gait disturbance, and urinary incontinence. Rarely, NPH can present with bulbar involvement, particularly with difficulty swallowing. Here, we present a case of NPH in a 75-year-old man who presented with an episode of choking and a recent onset of swallowing difficulties with a three-month history of ataxia and progressive memory loss. His CT scan revealed ventriculomegaly, which was consistent with the clinical presentation of NPH and was further confirmed by a normal opening pressure on the CSF tap. Furthermore, ventriculoperitoneal shunts showed a marked improvement in patients' dysphagia and the classical triad of NPH symptoms. Through this case report, we want to highlight that NPH can present as a difficulty in swallowing.

Efficacy and Safety of Sodium Glucose Cotransporter-2 (SGLT2) Inhibitors in Patients With Diabetes and Chronic Kidney Disease (CKD): A Meta-analysis of Randomized Control Trials.

The current meta-analysis aims to assess the efficacy and safety of sodium glucose cotransporter 2 (SGLT2) inhibitors in individuals with diabetes and chronic kidney disease (CKD). The current meta-analysis was conducted using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. A systematic search was conducted to identify all relevant studies related to the efficacy and safety of SGLT2 inhibitors in individuals with diabetes and CKD. The search was undertaken in PubMed, EMBASE, and Cochrane Library from January 2000 to September 2022. The primary efficacy outcome assessed in the current meta-analysis included major adverse cardiovascular events (MACE). Other efficacy outcomes included all-cause mortality and change in hemoglobin A1c (HbA1c) (%). Safety outcomes included serious adverse events, acute kidney injury, hypoglycemia, and hyperkalemia. In total 11 articles met the inclusion criteria and were included in the final analysis enrolling 27520 patients (14491 in the SGLT2 inhibitors and 13029 in the placebo group). The findings of this meta-analysis have shown that the risk of MACE and all-cause mortality was significantly lower in patients receiving SGLT2 inhibitors. Additionally, Hb1AC change was also significantly greater in SGLT2 inhibitors group. In relation to safety outcomes, serious adverse events, risk of acute kidney injury, and hyperkalemia were significantly lower in the SGLT2 inhibitors group. The SGLT2 inhibitors significantly decreased the risk of major cardiovascular events and all-cause mortality in patients with CKD and diabetes. Furthermore, SGLT2 inhibitor is also effective in reducing Hb1Ac levels in patients.

Determination of Alteration in Micromeritic Properties of a Solid Dispersion: Brunauer-Emmett-Teller Based Adsorption and Other Structured Approaches.

The present study is focused on the use of solid dispersion technology to triumph over the solubility-related problems of bexarotene which is currently used for treating various types of cancer and has shown potential inhibitory action on COVID-19 main protease and human ACE2 receptors. It is based on comparison of green locust bean gum and synthetic poloxamer as polymers using extensive mechanistic methods to explore the mechanism behind solubility enhancement and to find suitable concentration of drug to polymer ratio to prepare porous 3rd generation solid dispersion. The prepared solid dispersions were characterized using different studies like X-ray diffraction (XRD), thermal gravimetric analysis (TGA), scanning electron microscopy (SEM), Brunauer-Emmett-Teller (BET), differential scanning calorimetry (DSC), and particle size analysis in order to determine the exact changes occurred in the product which are responsible for enhancing solubility profiles of an insoluble drug. The results showed different profiles for particle size, solubility, dissolution rate, porosity, BET, and Langmuir specific surface area of prepared solid dispersions by using different polymers. In addition to the comparison of polymers, the BET analysis deeply explored the changes occurred in all dispersions when the concentration of polymer was increased. The optimized solid dispersion prepared with MLBG using lyophilization technique showed reduced particle size of 745.7±4.4 nm, utmost solubility of 63.97%, pore size of 211.597 Å, BET and Langmuir specific surface area of 5.6413 m2/g and 8.2757 m2/g, respectively.

Comparison of prophylactic phenylephrine and norepinephrine infusion on umbilical arterial pH and maternal blood pressure during spinal anaesthesia for caesarean delivery.

Background and Aims: Spinal anaesthesia induced maternal hypotension in parturients undergoing caesarean delivery may lead to neonatal acidosis and fall in umbilical artery pH. The aim of this study was to compare low dose norepinephrine infusion with phenylephrine to see the effect on umbilical arterial pH and maternal blood pressure during spinal anaesthesia for caesarean delivery. Methods: In a randomised, double-blind study, 60 parturients belonging to American Society of Anesthesiologists grade II, age 18-35 years with singleton term pregnancy were divided into the phenylephrine group and norepinephrine group. Participants received prophylactic phenylephrine and norepinephrine infusion after spinal anaesthesia till the delivery of the baby at a fixed rate of 50 µg/min and 2.5 µg/min, respectively. The primary outcome was umbilical artery pH. Neonatal Apgar score, incidence of bradycardia and hypotension, number of boluses of vasopressor required and reactive hypertension were also compared. Results: The umbilical arterial pH was comparable between the groups (p = 0.38). Apgar scores were comparable (p = 0.17). Incidence of bradycardia was higher in phenylephrine group without reaching statistical significance (43.3% vs. 20%, P = 0.052). Incidence of hypotension was more but not significant in norepinephrine group compared to phenylephrine group (16.7% vs. 10%, P = 0.44). Number of vasopressor boluses and reactive hypertension episodes were comparable between both groups (p = 0.09). Conclusion: Low dose (2.5 µg/min) intravenous infusion of norepinephrine is a suitable alternative to phenylephrine in the maintenance of umbilical arterial pH and maternal blood pressure.

Structural Insights in Mammalian Sialyltransferases and Fucosyltransferases: We Have Come a Long Way, but It Is Still a Long Way Down.

Mammalian cell surfaces are modified with complex arrays of glycans that play major roles in health and disease. Abnormal glycosylation is a hallmark of cancer; terminal sialic acid and fucose in particular have high levels in tumor cells, with positive implications for malignancy. Increased sialylation and fucosylation are due to the upregulation of a set of sialyltransferases (STs) and fucosyltransferases (FUTs), which are potential drug targets in cancer. In the past, several advances in glycostructural biology have been made with the determination of crystal structures of several important STs and FUTs in mammals. Additionally, how the independent evolution of STs and FUTs occurred with a limited set of global folds and the diverse modular ability of catalytic domains toward substrates has been elucidated. This review highlights advances in the understanding of the structural architecture, substrate binding interactions, and catalysis of STs and FUTs in mammals. While this general understanding is emerging, use of this information to design inhibitors of STs and FUTs will be helpful in providing further insights into their role in the manifestation of cancer and developing targeted therapeutics in cancer.

MR Venography in Gamma Knife Radiosurgery for Parasagittal Meningiomas: A Technical Note with the Rationale of Venous Protection and a Review of Literature.

Postradiosurgery symptomatic brain edema may be seen with parasagittal meningioma owing to its proximity to major venous sinuses and cortical veins. Venous preservation radiosurgery planning is less described. Here, we discuss a new method of tumor volume contouring on postcontrast magnetic resonance venogram (CEMRV) images safely excluding the adjacent cortical veins and sinuses. Six cases of parasagittal meningiomas where Gamma Knife radiosurgery was planned on CEMRV sequence were studied in detail. A double-contrast injection method was used to obtain CEMRV images. The differential contrast enhancement showed the displaced and compressed sinuses and cortical veins in the vicinity of meningioma. Tumor was contoured on both contrast magnetic resonance imaging (CEMRI) and MRV image for comparative analysis. 15 Gy at 50% marginal isodose was prescribed and quantitative assessment showed reduced exposure to the adjacent veins and sinuses on the MRV plan as compared to the CEMRI plan. All patients remain asymptomatic at a mean follow-up of 34.2 months. Postcontrast MRV is a simple sequence and can delineate the adjacent venous structures in parasagittal meningiomas. Tumor contouring directly on this sequence guides the surgeon to prescribe adequate radiation dose while sparing cortical veins and sinuses in radiosurgery planning.

Flowerpot method for rapid eye movement sleep deprivation does not induce stress as defined by elevated serum corticosterone level in rats.

Flowerpot method of rapid eye movement sleep (REMS) deprivation (REMSD) has been most extensively used in experiments to decipher the functions of REMS. The most common but serious criticism of this method has been presumed stress experienced by the experimental animals. The lack of systematic studies with appropriate controls to resolve this issue prompted this study. We have compared serum corticosterone levels as a marker of stress in male rats under REMSD by the flowerpot method and multiple types of control conditions. Additionally, to maintain consistency and uniformity of REMSD among groups, in the same rats, we estimated brain Na-K ATPase activity, which has been consistently reported to increase upon REMSD. The most effective method was one rat in single- or multiple-platforms set-up in a pool because it significantly increased Na-K ATPase activity without elevating serum corticosterone level. More than one rat in multiple platform set-up was ineffective and must be avoided. Also, large platform- and recovery-controls must be carried out simultaneously to rule out non-specific confounding effects.

Tmprss2 specific miRNAs as promising regulators for SARS-CoV-2 entry checkpoint.

Tmprss2 is an emerging molecular target which guides cellular infections of SARS-CoV-2, has been earmarked for interventions against the viral pathologies. The study aims to computationally screen and identifies potential miRNAs, following in vitro experimental validation of miRNA-mediated suppression of Tmprss2 for early prevention of COVID-19. Pool of 163 miRNAs, scrutinized for Tmprss2 binding with three miRNA prediction algorithms, ensued 11 common miRNAs. Further, computational negative energies for association, corroborated miRNA-Tmprss2 interactions, whereas three miRNAs (hsa-miR-214, hsa-miR-98 and hsa-miR-32) based on probability scores ≥0.8 and accessibility to Tmprss2 target have been selected in the Sfold tool. Transfection of miRNA(s) in the Caco-2 cells, quantitatively estimated differential expression, confirming silencing of Tmprss2 with maximum gene suppression by hsa-miR-32 employing novel promising role in CoV-2 pathogenesis. The exalted binding of miRNAs to Tmprss2 and suppression of later advocates their utility as molecular tools for prevention of SARS-CoV-2 viral transmission and replication in humans.

Individual, interpersonal and economic challenges of underemployment in the wake of COVID-19.

BACKGROUND: COVID-19 has caused economic slowdown all across the globe. It results in job loss on the one hand and less wages, increased working hours, overqualified employees and part time jobs on the other hand. Low demand of labour and a huge availability of work force will put many in a disadvantageous position, where they will have to compromise with the circumstances by being underemployed. Cabinet decisions, by some Indian states like Uttar Pradesh, to suspend the labour laws related to minimum wage, bonus, working hours and other employee benefits will put workers in a highly disadvantageous position of being underemployed. This may lead to many socio-economic, psychological and health-related implications. OBJECTIVE: The aim of this paper is to provide a comprehensive review on the concept of underemployment, its types and consequences. METHODOLOGY: A critical and constructive analysis of the literature was performed. RESULTS: The findings reveal that if employment does not provide workers with proper opportunities to use their education, time, skills and expertise, it can create stressful situations in workers' lives. CONCLUSION: The problem needs both a diagnosis and robust treatment in order to have better outcomes at the individual, organisational and national levels.

Noradrenaline Acting on Alpha1 Adrenoceptor as well as by Chelating Iron Reduces Oxidative Burden on the Brain: Implications With Rapid Eye Movement Sleep.

The noradrenaline (NA) level in the brain is reduced during rapid eye movement sleep (REMS). However, upon REMS deprivation (REMSD) its level is elevated, which induces apoptosis and the degeneration of neurons in the brain. In contrast, isolated studies have reported that NA possesses an anti-oxidant property, while REMSD reduces lipid peroxidation (LP) and reactive oxygen species (ROS). We argued that an optimum level of NA is likely to play a physiologically beneficial role. To resolve the contradiction and for a better understanding of the role of NA in the brain, we estimated LP and ROS levels in synaptosomes prepared from the brains of control and REMS deprived rats with or without in vivo treatment with either α1-adrenoceptor (AR) antagonist, prazosin (PRZ) or α2-AR agonist, clonidine (CLN). REMSD significantly reduced LP and ROS in synaptosomes; while the effect on LP was ameliorated by both PRZ and CLN; ROS was prevented by CLN only. Thereafter, we evaluated in vitro the effects of NA, vitamin E (Vit E), vitamin C (Vit C), and desferrioxamine (DFX, iron chelator) in modulating hydrogen peroxide (H2O2)-induced LP and ROS in rat brain synaptosomes, Neuro2a, and C6 cells. We observed that NA prevented ROS generation by chelating iron (inhibiting a Fenton reaction). Also, interestingly, a lower dose of NA protected the neurons and glia, while a higher dose damaged the neurons and glia. These in vitro and in vivo results are complementary and support our contention. Based on the findings, we propose that REMS maintains an optimum level of NA in the brain (an antioxidant compromised organ) to protect the latter from continuous oxidative onslaught.

Impact of health education on knowledge, attitude, practices and glycemic control in type 2 diabetes mellitus.

BACKGROUND: Effective health education consequently improves knowledge, attitude, and practices (KAP) leading to better glycemic control and is widely accepted as an integral part of comprehensive diabetes care for affected individuals and their families in primary care settings. AIMS: To assess the impact of health education on knowledge, attitude, practices, and glycemic control in type 2 diabetes mellitus patients. SETTINGS AND DESIGN: This case control study was conducted in the department of Medicine of a tertiary care teaching hospital. METHODS: The study was conducted on 100 diabetic subjects aged more than 40 years comprising of 50 cases and 50 controls. Cases were given education on their disease, drugs, dietary, and lifestyle modifications along with patient education leaflet at baseline and at first follow-up, while controls received neither of these. Subjects were assessed for KAP by administering KAP questionnaire and for glycemic control by measuring glycated hemoglobin (HbA1C) at baseline and at the end of the study. RESULTS: In this study, end mean knowledge, attitude, practice, and KAP SUM scores of cases (10.28 ± 1.78, 3.46 ± 0.93, 3.14 ± 0.86, and 16.82 ± 3.40, respectively) showed significant increase from the baseline (3.86 ± 0.93, 1.00 ± 0.83, 0.40 ± 0.64, and 5.26 ± 2.10, respectively) compared to controls, accompanied by significant reduction in HbA1C of cases at the end of the study compared to the controls. CONCLUSIONS: Effective health education improves knowledge, attitude, and practices, particularly with regard to lifestyle modifications and dietary management, culminating into better glycemic control that can slow down the progression of diabetes and prevent downstream complications.

Quantification of desmosine and isodesmosine using MALDI-ion trap tandem mass spectrometry.

Desmosine (Des) and isodesmosine (Isodes), cross-linking amino acids in the biomolecule elastin, may be used as biomarkers for various pathological conditions associated with elastin degradation. The current study presents a novel approach to quantify Des and Isodes using matrix-assisted laser desorption ionization (MALDI)-tandem mass spectrometry (MS2) in a linear ion trap coupled to a vacuum MALDI source. MALDI-MS2 analyses of Des and Isodes are performed using stable-isotope-labeled desmosine d4 (labeled-Des) as an internal standard in different biological fluids, such as urine and serum. The method demonstrated linearity over two orders of magnitude with a detection limit of 0.02 ng/µL in both urine and serum without enrichment prior to mass spectrometry, and relative standard deviation of < 5%. The method is used to evaluate the time-dependent degradation of Des upon UV irradiation (254 nm) and found to be consistent with quantification by 1H NMR. This is the first characterized MALDI-MS2 method for quantification of Des and Isodes and illustrates the potential of MALDI-ion trap MS2 for effective quantification of biomolecules. The reported method represents improvement over current liquid chromatography-based methods with respect to analysis time and solvent consumption, while maintaining similar analytical characteristics. Graphical abstract ᅟ.

Comparative Evaluation of Anti Microbial effects of Triple Antibiotic Paste and Amox and its derivatives against E. Faecalis: An in vitro study.

BACKGROUND: Enterococcus faecalis is a microorganism commonly detected in asymptomatic, persistent endodontic infections. Triple antibiotic paste has stood the test of time as a proven antibiotic combination against E. Faecalis. However, problems with this include staining of teeth and standardization of the preparation. Thus, the search for better alternatives and better preparation techniques is still on. AIM: To observe the potential of combinations [(Amoxicillin+ Metronidazole, Amoxicillin Clavulanic Acid + Metronidazole; Amoxicillin and Cloxacillin + Metronidazole)] over Triple Antibiotic Paste. MATERIAL AND METHODS: Fifty single rooted teeth free from dental caries were selected for the study. Teeth were cut at equal distance from root apex (13mm from apex) with sterile diamond disk and straight hand piece for standardization of root length. The opening of root canal was enlarged with engine driven pro-taper files. To remove the organic and inorganic debris, canal was cleaned with 17% EDTA followed by 2.5% NaOCl for 5min. Distilled water irrigation was done for 5 min to remove any traces of used chemical and then sterilized in autoclave at 1200c for 15 min. at 15 lbs pressure. Bacteria cultured on blood agar plate and at the same time fresh antibiotic combinations were made and placed in the root canals, then incubated in the incubator, under sterile conditions and observed at 24hrs, 48hr and 72hrs. RESULTS: The largest inhibition zones were observed for the Triple Antibiotic Paste, followed by Amoxicillin and Clavulanic acid + Metronidazole group however, the clearest zones were achieved with Amoxicillin and Clavlunic acid + Metronidazole group and the smallest for Amoxicillin and Metronidazole group. CONCLUSIONS: The results suggest that though Triple antibiotic showed the maximum inhibition, Amoxicillin and Clavulanic acid combination along with Metronidazole gave the most reliable results. Further studies using the different combinations and different concentrations along with different methods of increasing the shelf life of such medications can be undertaken. Key words:Enterococcus faecalis, Triple Antibiotic Paste, Amoxcillin, Clavulanic acid.

Endometrial evaluation by ultrasonography, hysteroscopy and histopathology in cases of breast carcinoma on Tamoxifen therapy.

INTRODUCTION: Tamoxifen, a nonsteroidal antiestrogenic agent, is used widely as adjunctive therapy for women with breast cancer. Most studies have found that the increased relative risk of developing endometrial cancer for women taking Tamoxifen is two to three times higher than that of an age-matched population. So we designed this study to assess the endometrial status in patients taking Tamoxifen for breast carcinoma. MATERIAL AND METHODS: The study was conducted at Govt. Medical College and Rajindra Hospital, Patiala, India. A total of 50 patients of Ca Breast taking Tamoxifen were selected as per study criterion and TVS performed. If endometrial thickness was more than 5 mm hysteroscopy and endometrial HPE was done and data analysed. RESULTS: On ultrasonography 35 patients (70%) had an endometrial thickness up to 5 mm. 15 patients (30%) had an endometrial thickness more than 5mm. Out of these, 11 patients, i.e. 22% of total, had an endometrial thickness of 5.1 to 10 mm and 2 patients, i.e. 4% of total had an endometrial thickness of more than 20 mm. Hysteroscopy was done on 11 patients. Out of these 8 patients had a normal hysteroscopic appearance whereas 3 patients had an abnormal hysteroscopic picture. Endometrial HPE of these 11 patients revealed 2 patients had secretory changes, 1 had polyp change, 1 had atrophic endometrium, 3 had simple endometrial hyperplasia, 1 had endometrial adenocarcinoma and 4 patients were reported to have scanty curetting. CONCLUSION: The duration of Tamoxifen therapy turned out to have a relationship with the incidence of endometrial carcinoma (P < 0.0001). Also, a relationship was observed between the duration of Tamoxifen therapy and symptom status of the patients (P < 0.0001). This correlation did not extend to duration of Tamoxifen therapy and endometrial thickness. (P = 0.190). This correlation did not extend to duration of Tamoxifen therapy and endometrial thickness. (P = 0.190).