Spectroscopic characterization and pharmacokinetic evaluation of amorphous solid dispersions of glibenclamide for bioavailability enhancement in Wistar rats.

Oral bioavailability of glibenclamide (Glb) was appreciably improved by the formation of an amorphous solid dispersion with Poloxamer-188 (P-188). Poloxamer-188 substantially enhanced the solubility and thereby the dissolution rate of the biopharmaceutics classification system (BCS) class II drug Glb and simultaneously exhibited a better stabilizing effect of the amorphous solid dispersion prepared by the solvent evaporation method. The physical state of the dispersed Glb in the polymeric matrix was characterized by differential scanning calorimetry, X-ray diffraction, scanning electron microscope and Fourier transform infrared studies. In vitro drug release in buffer (pH 7.2) revealed that the amorphous solid dispersion at a Glb-P-188 ratio of 1:6 (SDE4) improved the dissolution of Glb by 90% within 3 h. A pharmacokinetic study of the solid dispersion formulation SDE4 in Wistar rats showed that the oral bioavailability of the drug was greatly increased as compared with the market tablet formulation, Daonil®. The formulation SDE4 resulted in an AUC0-24h ~2-fold higher. The SDE4 formulation was found to be stable during the study period of 6 months.

Optimizing the fabrication of electrospun nanofibers of prochlorperazine for enhanced dissolution and permeation properties.

Despite being an approved antiemetic for more than five decades, the clinical usefulness of prochlorperazine is limited by its low solubility and inconsistent absorption in the gastrointestinal tract, which presents challenges for nanotherapeutic interventions. Here, we report the preparation of a highly soluble and permeable nanofiber formulation of prochlorperazine using the Quality-by-Design approach. The final nanofiber formulation with drug entrapment of 88.02 ± 1.14 % was obtained at 20.0 kV, with a flow rate of 0.5 ml/h and tip-to-collector distance of 19.9 cm. Physio-mechanical properties, such as thickness (0.42 ± 0.02 mm), pH resistance (7.04 ± 0.08), folding endurance (54 ± 5), and tensile strength (0.244 ± 0.02 N.mm-2), were appropriate for packaging and application to oromucosal surfaces. The content uniformity (93.48-106.63 %) and weight variation (<1.8 mg) of the optimal nanofiber formulation were within the permissible limits prescribed for orodispersible films. Microscopical investigations confirm a randomly deposited and dense network of woven nanofibers with an average diameter of 363 ± 5.66 nm. The drug particles were embedded homogeneously on the fiber in the nanoform (4.27 ± 1.34 nm). The spectral analysis using TEM-EDS shows diffraction peaks of sulfur and chlorine, the elemental constituents of prochlorperazine. The drug was amorphized in the nanofiber formulation, as led by the decline of the crystallinity index from 87.25 % to 7.93 % due to electrostatic destabilization and flash evaporation of the solvent. The enthalpy of fusion values of the drug in the nanofiber mat decreased significantly to 23.6 J/g compared to its pristine form, which exhibits a value of 260.7 J/g. The nanofibers were biocompatible with oral mucosal cells, and there were no signs of mucosal irritation compared to 1 % sodium lauryl sulfate. The fiber mats rapidly disintegrated within <1 s and released ≈91.49 ± 2.1 % of the drug within 2 min, almost 2-fold compared to the commercial Stemetil MD® tablets. Similarly, the cumulative amount of the drug permeated across the unit area of the oromucosal membrane was remarkably high (31.28 ± 1.30 µg) compared to 10.17 ± 1.11 µg and 13.10 ± 1.79 µg from the cast film and drug suspension. Our results revealed these nanofiber formulations have the potential to be fast-dissolving oromucosal delivery systems, which can result in enhanced bioavailability with an early onset of action due to rapid disintegration, dissolution, and permeation.

Validation and development of RP-HPLC method for quantification of glibenclamide in rat plasma and its application to pharmacokinetic studies in wistar rats.

Herein, we report a novel, simple, specific, accurate and cost-friendly validated reverse phase-high performance liquid chromatographic (RP-HPLC) method for the quantification of second generation sulphonylurea based antidiabetic drug, glibenclamide (GLB) in rat plasma and its application to calculate pharmacokinetic parameters in wistar rats. The internal standard used was flufenamic acid. The chromatographic separation was conducted on C18 column (250 mm × 4.6 mm x 5 µm, Agilent-Zorbax, SB) using isocratic elution with mobile phase containing Acetonitrile: Water (1:1; v/v) pH adjusted to 4.0 with 0.03 % glacial acetic acid and detected by photo-diode array as detector. Calibration curves made in the rat plasma were linear in the range of 50-1200 ng/ml with r2 = 0.998. The LLOQ was 40 ng/ml. This method was effectively applied for pharmacokinetic studies of Glibenclamide following administration through oral route as solid dispersion formulation to Wistar rats. Several methods are available in the literature which can be employed for the quantification of Glibenclamide but such methods are tedious, provide lower sensitivity, less simultaneous resolution and are time-consuming. Therefore the present methods suits best for the quantification of Glibenclamide from Wistar rats.

A Rare Incidence of Hepatocellular Carcinoma With Tumor Thrombus Extending to the Right Heart.

Hepatocellular carcinoma (HCC) is a leading cause of cancer-related deaths and the sixth most commonly diagnosed cancer worldwide due to several common risk factors, including hepatitis C virus (HCV), hepatitis B virus (HBV), and other causes of cirrhosis. In HCC, intrahepatic vascular invasion and a tumor thrombus are commonly observed. However, the extrahepatic spread of the tumor thrombus to the heart via the portal vein, hepatic vein, and inferior vena cava (IVC) is rarely reported and is considered a poor prognostic factor. In addition, rarely, there is a risk of cor pulmonale and thromboembolism of the pulmonary vessels. Our patient also presented with this rare complication of HCC. Our patient's clinical presentation was bilateral pedal edema, moderate ascites, and abdominal discomfort with raised jugular venous pressure. These signs and symptoms are related to an impairment of the right heart caused by intracardiac tumor thrombus metastasis, leading to diastolic dysfunction. Based on these findings, echocardiography and abdominal computed tomography (CT) scan were performed with the definitive diagnosis of hepatocellular carcinoma with tumor thrombus metastases in the hepatic vein, inferior vena cava, and right atrium. The management team agreed on a conservative treatment plan based on the advanced stage of the disease and the high risk associated with aggressive treatment modalities. Unfortunately, on day 7 of admission, the patient died from a possible pulmonary embolism that led to cardiopulmonary arrest. This case underscores the importance of screening patients with a high HCC tumor burden with abdominal ultrasound and echocardiography for early detection and timely management.

Simultaneous quantification of losartan potassium and its active metabolite, EXP3174, in rabbit plasma by validated HPLC-PDA.

Herein, we report a novel, accurate and cost-effective validated analytical method for the quantification of losartan potassium and its active metabolite, EXP 3174, in rabbit plasma by reversed-phase high-performance liquid chromatography. Valsartan was used as an internal standard. The method was validated as per International Conference on Harmonization guidelines. The analytes were extracted in rabbit plasma using liquid-liquid extraction technique and analyzed at 247 nm after separation through a reverse-phase C18 column. The isocratic mobile phase used is a mixture of acetonitrile, water and glacial acetic acid in the ratio of 60:40:1 v/v/v maintained at pH 3.4. All calibration curves showed a good linear relationship (r > 0.995) within the test range. Precision was evaluated by intra- and interday tests with RSDs <1.91% and accuracy showed validated recoveries of 86.20-101.11%. Based on our results, the developed method features good quantification parameters and can serve as an effective quality control method for the standardization of drugs.

Egg Consumption in the Context of Plant-Based Diets and Diet Quality in Adults at Risk for Type 2 Diabetes: A Randomized Single Blind Cross-over Controlled Trial.

Lifestyle changes that emphasis on plant-based diets (PBD) are typically recommended for those at risk for type 2 diabetes mellitus (T2DM) to mitigate their cardo-metabolic risk. We examined the impact of the inclusion of eggs compared with their exclusion from PBD on diet quality among adults at risk for T2DM.This was a randomized, controlled, single-blind, crossover trial of 35 adults (mean age 60.7 years; 25 women, 10 men; 34 Caucasians, 1 African-American) at risk for T2DM (i.e., pre- diabetes or metabolic syndrome) assigned to one of two possible sequence permutations of two treatments (PBD with eggs and exclusively PBD), with a 4-week washout period. Participants received dietary counseling from a dietitian to exclude or to include 2 eggs daily in the context of PBD for a 6-week period. Diet quality was assessed using the Healthy Eating Index 2015 (HEI-2015) at baseline and 6 weeks.Compared with the exclusion of eggs, the inclusion of eggs in the context of PBD improved the diet quality score for intake of total protein foods (1.0 ± 1.1 vs. -0.4 ± 1.0; p <.0001); seafood and plant proteins (0.2 ± 1.2 vs. -0.4 ± 1.1; p = 0.0338); and fatty acids (0.8 ± 2.5 vs. -0.7 ± 2.7; p = 0.0260). Overall diet quality score depreciated with the adoption of exclusively PBD without eggs (-3.1 ± 8.3; p = 0.0411), while it was unaffected with the adoption of a PBD with the inclusion of eggs (-0.6 ± 7.9; p = 0.6892).Eggs could be used as an adjuvant to enhance the diet quality among those at risk for T2DM who adopt plant-based dietary patterns.

Global Effect of COVID-19 Pandemic on Cancer Patients and its Treatment: A Systematic Review.

Background: At a global level, the COVID-19 disease outbreak has had a major impact on health services and has induced disruption in routine care of health institutions, exposing cancer patients to severe risks. To provide uninterrupted tumor treatment throughout a pandemic lockdown is a major obstacle. Coronavirus disease (COVID-19) and its causative virus, SARS-CoV-2, stance considerable challenges for the management of oncology patients. COVID-19 presents particularly severe respiratory and systemic infection in aging and immunosuppressed individuals, including patients with cancer. Objective: In the present review, we focused on emergent evidence from cancer sufferers that have been contaminated with COVID-19 and cancer patients who were at higher risk of severe COVID-19, and indicates that anticancer treatment may either rise COVID-19 susceptibility or have a duple therapeutic impact on cancer as well as COVID-19; moreover, how SARS-CoV-2 infection impacts cancer cells. Also, to assess the global effect of the COVID-19 disease outbreak on cancer and its treatment. Methods: A literature survey was conducted using PubMed, Web of Science (WOS), Embase, Cochrane Library, China National Knowledge Infrastructure (CNKI), and VIral Protein domain DataBase (VIP DB) between Dec 1, 2019 and Sep 23, 2021, for studies on anticancer treatments in patients with COVID-19. The characteristics of the patients, treatment types, mortality, and other additional outcomes were extracted and pooled for synthesis. Results: This disease has a huge effect on sufferers who have cancer(s). Sufferers of COVID-19 have a greater percentage of tumor diagnoses than the rest of the population. Likewise, cancer and highest proportion is lung cancer sufferers are more susceptible to COVID-19 constriction than the rest of the population. Conclusion: Sufferers who have both COVID-19 and tumor have a considerably elevated death risk than single COVID-19 positive patients overall. During the COVID-19 pandemic, there was a reduction in the screening of cancer and detection, and also deferral of routine therapies, which may contribute to an increase in cancer mortality there in future.

Experience of psychologists in the delivery of cognitive behaviour therapy in a non-western culture for treatment of substance abuse: a qualitative study.

BACKGROUND: Psychotherapy is the preferred form of treatment for psychological disorders worldwide. Cognitive behaviour therapy (CBT) is one of the most widely used psychotherapies due to its proven efficacy for psychological disorders, including substance abuse. However, CBT was developed in the West according to the culture of developed countries. Therefore, it requires cross-cultural adaptation for non-Western countries. Pakistan is one of the developing non-Western countries where substance use disorders are increasing at an alarming rate. Despite the proven efficacy of CBT for substance use disorders, there is a dearth of its utilization in Pakistan. Therefore, in the present study, in-depth qualitative interviews were conducted with CBT practitioners in Pakistan to understand barriers and challenges in this regard. The study was a part of a broader project aimed at cultural adaptation of CBT for people with substance use disorders (SUDs) in Pakistan. METHODS: In-depth qualitative interviews were conducted with CBT practitioners (N = 8) working in rehabilitation centres and hospitals in Islamabad, Pakistan. Thematic content analysis was conducted to develop core themes from the data. RESULTS: CBT for SUDs requires some adjustments according to Pakistani culture for successful utilization. The challenges in providing CBT for SUDs revolved around three main themes, i.e., the mental health system, societal practices, and therapeutic issues, and 10 subthemes. CONCLUSION: In order to utilize the benefits of CBT for SUDs in Pakistan, cultural adaptation is necessary as an initial step. However, its delivery requires stringent modifications in the health care system to address these challenges.

Formulation and Pharmacokinetic Evaluation of Ethyl Cellulose/HPMC-Based Oral Expandable Sustained Release Dosage of Losartan Potassium.

Prolonged retention of losartan potassium in the upper gastrointestinal tract is anticipated to increase its absorption and exposure to CYP450 enzyme subfamilies, undertaking its conversion to more potent (10-40 times) active metabolite, losartan carboxylic acid (LCA). Consistent with this, hydroxypropyl methylcellulose K4M/ethyl cellulose-based novel expandable films (EFs) containing losartan potassium (LP) suitable for prolonged retention in the stomach were developed. The films were prepared by solvent casting method. USP type II dissolution apparatus (0.1 N HCl, 37°C, 100 rpm) was used to perform the dissolution testing (drug release, unfolding behavior, film integrity, erosion, and water uptake) of the films. In vivo pharmacokinetic studies were carried out in rabbits. An HPLC-UV method was used for the quantification of the drug and its active metabolite in plasma. These folded films placed inside hard gelatin capsule shells unfolded to full dimensions in dissolution medium and provided sustained drug release throughout 12 h. The plasma drug concentration-time curves obtained from the in vivo studies were used to determine pharmacokinetic parameters, such as area under the plasma drug concentration-time curve (AUC), area under first moment curve (AUMC), mean residence time (MRT), Cmax, Tmax, t1/2, ke, and Fr in comparison with that of the market formulation, Cozaar®. The novel EFs significantly changed the pharmacokinetic parameters of the drug and its active metabolite. The apparent elimination rate constant (ke) significantly decreased, while MRT and elimination half-life (t1/2) increased in both cases. The relative bioavailabilities (Fr) of both LP and E3174 using the novel formulation were higher than that of Cozaar®.

Multiple-reaction monitoring (MRM) LC-MS/MS quantitation of venlafaxine and its O-desmethyl metabolite for a preclinical pharmacokinetic study in rabbits.

Preclinical pharmacokinetic (PK) studies in animal models during the formulation development phase give preliminary evidence and near clear picture of the PK behavior of drug and/or its dosage forms before clinical studies on humans and help in the tailoring of the dosage form according to the expected and requisite clinical behavior. The present work reports a first of its kind preclinical PK study on extended-release (ER) solid oral dosage forms of venlafaxine (VEN) in New Zealand White rabbits. The VEN is a highly prescribed and one of the safest and most effective therapeutic agents used in the treatment of different types of depression disorders worldwide. The multiple-reaction monitoring (MRM) LC-MS/MS method developed for this purpose demonstrated enough reliability in simultaneously quantitating VEN and its equipotent metabolite O-desmethylvenlafaxine (ODV) in rabbit plasma. The method described uses solid-phase extraction for sample preparation followed by an ultrafast LC-MS/MS analysis. The chromatographic separation was achieved isocratically with a predominantly polar mobile phase by employing RPLC. The triple quadrupole LC/MS/MS system operated in MRM mode used an ESI probe as an ion source in positive polarity. The validation results are within the permissible limits of US FDA recommendations and acceptance criteria for bioanalytical method validation.

Utilization of Artificial Intelligence in Disease Prevention: Diagnosis, Treatment, and Implications for the Healthcare Workforce.

Artificial intelligence (AI) has been described as one of the extremely effective and promising scientific tools available to mankind. AI and its associated innovations are becoming more popular in industry and culture, and they are starting to show up in healthcare. Numerous facets of healthcare, as well as regulatory procedures within providers, payers, and pharmaceutical companies, may be transformed by these innovations. As a result, the purpose of this review is to identify the potential machine learning applications in the field of infectious diseases and the general healthcare system. The literature on this topic was extracted from various databases, such as Google, Google Scholar, Pubmed, Scopus, and Web of Science. The articles having important information were selected for this review. The most challenging task for AI in such healthcare sectors is to sustain its adoption in daily clinical practice, regardless of whether the programs are scalable enough to be useful. Based on the summarized data, it has been concluded that AI can assist healthcare staff in expanding their knowledge, allowing them to spend more time providing direct patient care and reducing weariness. Overall, we might conclude that the future of "conventional medicine" is closer than we realize, with patients seeing a computer first and subsequently a doctor.

Scavenging of Organic Pollutant and Fuel Generation through Cost-Effective and Abundantly Accessible Rust: A Theoretical Support with DFT Simulations.

Waste management and energy generation are the foremost concerns due to their direct relationship with biological species and the environment. Herein, we report the utilization of iron rust (inorganic pollutant) as a photocatalyst for the photodegradation of methylene blue (MB) dye (organic pollutant) under visible light (economic) and water oxidation (energy generation). Iron rust was collected from metallic pipes and calcined in the furnace at 700 °C for 3 h to remove the moisture/volatile content. The uncalcined and calcined rust NPs are characterized through scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS), Fourier-transform infrared (FTIR) analysis, X-ray Diffraction (XRD), and thermogravimetric analysis (TGA). The morphological study illustrated that the shape of uncalcined and calcined iron rust is spongy, porous, and agglomerated. The XRD and DLS particle sizes are in a few hundred nanometers range. The photodegradation (PD) investigation shows that calcined rust NPs are potent for the PD of modeled MB, and the degradation efficiency was about 94% in a very short time of 11 min. The photoelectrochemical (PEC) measurements revealed that calcined rust NPs are more active than uncalcined rust under simulated 1 SUN illumination with the respective photocurrent densities of ~0.40 and ~0.32 mA/cm2. The density functional theory simulations show the chemisorption of dye molecules over the catalyst surface, which evinces the high catalytic activity of the catalyst. These results demonstrate that cheaper and abundantly available rust can be useful for environmental and energy applications.

Effectiveness and safety profiling of zolpidem and acupressure in CKD associated pruritus: An interventional study.

BACKGROUND: Chronic kidney disease (CKD)-associated pruritus (CKD-aP) contributes to poor quality of life, including reduced sleep quality and poor sleep quality is a source of patient stress and is linked to lower health-related quality of life. This study aimed to investigate the effectiveness of zolpidem 10 mg and acupressure therapy on foot acupoints to improve the sleep quality and overall quality of life among hemodialysis patients suffering from CKD-aP. METHOD: A multicenter, prospective, randomized, parallel-design, open label interventional study to estimate the effectiveness of zolpidem (10 mg) oral tablets versus acupressure on sleep quality and quality of life in patients with CKD-aP on hemodialysis. A total of 58 hemodialysis patients having sleep disturbance due to CKD-aP completed the entire 8-week follow-up. The patients were divided into a control (acupressure) group of 28 patients and an intervention (zolpidem) group of 30 patients. RESULTS: A total of 58 patients having CKD-aP and sleep disturbance were recruited. In the control group there was a reduction in the PSQI score with a mean ±â€ŠSD from 12.28 ±â€Š3.59 to 9.25 ±â€Š3.99, while in the intervention group the reduction in PSQI score with a mean ±â€ŠSD was from 14.73 ±â€Š4.14 to 10.03 ±â€Š4.04 from baseline to endpoint. However, the EQ5D index score and EQ-visual analogue scale (VAS) at baseline for the control group with a mean ±â€ŠSD was 0.49 ±â€Š0.30 and 50.17 ±â€Š8.65, respectively, while for the intervention group the values were 0.62 ±â€Š0.26 and 47.17 ±â€Š5.82, respectively. The mean EQ5D index score in the control group improved from 0.49 ±â€Š0.30 to 0.53 ±â€Š0.30, but in the intervention group there was no statistical improvement in mean EQ5D index score from 0.62 ±â€Š0.26 to 0.62 ±â€Š0.27 from baseline to week 8. The EQ 5D improved in both groups and the EQ-VAS score was 2.67 points higher at week 8 as compared to baseline in the control group, while in the intervention group the score was 3.33 points higher at week 8 as compared to baseline. Comparing with baseline, the PSQI scores were significantly reduced after week 4 and week 8 (P =  < .001). Furthermore, at the end of the study, the PSQI scores were significantly higher in the control as compared to the intervention group (P = .012). CONCLUSION: An improvement in sleep quality and quality of life among CKD-aP patients on hemodialysis has been observed in both the control and intervention groups. Zolpidem and acupressure safety profiling showed no severe adverse effect other that drowsiness, nausea and daytime sleeping already reported in literature of zolpidem.

Morpho-molecular identification and first report of Fusarium equiseti in causing chilli wilt from Kashmir (Northern Himalayas).

Chilli (Capsicum annuum L.) is one of the most significant vegetable and spice crop. Wilt caused by Fusarium Sp. has emerged as a serious problem in chilli production. Internal transcribed spacer (ITS) region is widely used as a DNA barcoding marker to characterize the diversity and composition of Fusarium communities. ITS regions are heavily used in both molecular methods and ecological studies of fungi, because of its high degree of interspecific variability, conserved primer sites and multiple copy nature in the genome. In the present study we focused on morphological and molecular characterization of pathogen causing chilli wilt. Chilli plants were collected from four districts of Kashmir valley of Himalayan region. Pathogens were isolated from infected root and stem of the plants. Isolated pathogens were subjected to DNA extraction and PCR amplification. The amplified product was sequenced and three different wilt causing fungal isolates were obtained which are reported in the current investigation. In addition to Fusarium oxysporum and Fusarium solani, a new fungal species was found in association with the chilli wilt in Kashmir valley viz., Fusarium equiseti that has never been reported before from this region. The studies were confirmed by pathogenicity test and re-confirmation by DNA barcoding.

Green Synthesis, Spectroscopic Characterization and Biomedical Applications of Carbon Nanotubes.

Carbon nanotubes are nano-sized cylindrical chicken wire-like structures made of carbon atoms. Carbon nanotubes have applications in electronics, energy storage, electromagnetic devices, environmental remediation and medicine as well. The biomedical applications of carbon nanotubes can be owed to features like low toxicity, non-immunogenicity, high in vivo stability and rapid cell entry. Carbon nanotubes have a great prospect in the treatment of diseases through diagnostic as well as therapeutic approaches. These nanostructures are interesting carriers for delivery and translocation of therapeutic molecules e.g. proteins, peptides, nucleic acids, drugs, etc. to various organs like the brain, lungs, liver, and pancreas. Commonly used methods to synthesize carbon nanotubes are arc discharge, chemical vapor deposition, pyrolysis, laser ablation etc. These methods have many disadvantages such as operation at high temperature, use of chemical catalysts, prolonged synthesis time and inclusion of toxic metallic particles in the final product requiring additional purification processes. In order to avoid these setbacks, various green chemistry-based synthetic methods have been devised, e.g., those involving interfacial polymerization, supercritical carbon dioxide drying, plant extract assisted synthesis, water- assisted synthesis, etc. This review will provide a thorough outlook of the eco-friendly synthesis of carbon nanotubes reported in the literature and their biomedical applications. Besides, the most commonly used spectroscopic techniques used for the characterization of carbon nanotubes are also discussed.

Determinants Of Bowel Resection In Childhood Intussusception.

BACKGROUND: Intussusception is an acute surgical emergency and one of the most common causes of acute abdomen in early paediatric age group with peak incidence between 3 months to 3 years. It is considered the most common cause of intestinal obstruction in first two years of life. Early diagnosis and management may prevent major complications of bowel ischemia and necrosis thus avoiding morbidity and mortality. Our objective was to evaluate the risk factors or determinants of bowel resection in childhood intussusception. METHODS: It is a prospective, descriptive and cross-sectional study in which 102 consecutive cases of intussusception presenting to The Children's Hospital PIMS, from January 2018 to May 2019, were included. RESULTS: Total 102 patients were included in the study. Seventy-three (71.6%) were male and 29 (28.4%) were female. Mean age at presentation was 16.30 months (ranged from 1 day to 9 years). Mean duration of symptoms was 3.1 days. 52.9% (53/102) patients had mass palpable on abdominal examination while 22.5% (23/102) had palpable mass on digital rectal examination (DRE). Mean haemoglobin at presentation was 10.2 g/dL (min 4g/dL, max 26g/dL) and mean haematocrit was 32.6%. The most common type of intussusception was ileo-colic (57.8%) followed by ileo-ileal (21.6%). 36.3% (37/102) patients had to undergo bowel resection due to bowel gangrene/necrosis while in 66 (64.7%) patients had manual reduction. Statistically significant relationship was found between resection and duration of symptoms more than 24 hours (p=0.01, CI=95% OR= 6.24), haemoglobin less than 10g/dL (p=0.022, CI=95% OR=2.78) and haematocrit less than 30% (p=0.028, CI=95% OR=2.63). CONCLUSIONS: Prevention of anaemia and early presentation to a facility where trained paediatric surgeons are available can reduce the chances of bowel resection in childhood intussusception.

Thyrostroma carpophilum insertional mutagenesis: A step towards understanding its pathogenicity mechanism.

Thyrostroma carpophilum, a causal agent of shot hole disease of stone fruits, cause severe loss in economically important fruit crops of Kashmir. Understanding its pathogenesis at molecular level will aid in devising a better management strategy. In this study, we optimized Agrobacterium tumefaciens mediated transformation (ATMT) conditions for T. carpophilum using PBIF2-EGFP construct. Using this protocol, we obtained 328 positive transformants per 104 spores and subsequent sub-culturing of transformants on selective and non-selective media resulted in stable T-DNA integration. Southern blot analysis revealed that most of the transformants embodied single T-DNA integration. Using this method, we obtained a small-scale transformant library (2050 transformants). Among this pool, we tested 1005 transformants for their pathogenicity; out of which 185 showed complete pathogenicity loss, 35 displayed reduced virulence and 785 were pathogenically similar to wild type. Out of this experimental stock, three transformants from each category were randomly selected to dissect the infection assay. The findings deciphered that transformants with complete pathogenicity loss failed to penetrate the host tissue and a few transformants failed to sporulate in laboratory. Transformants from reduced category could not form appressorium and occasionally sporulated. Transformants similar to wild type were morphologically and pathogenically similar to wild type because of un-alteration in their modus operandi. Our work provides a new platform to understand the pathogenicity mechanism of T. carpophilum. The optimized ATMT protocol will help in developing large transformant library that can help to identify the virulence arsenals necessary for the pathogen to cause disease.

Experimental Protocol for Induction of Transgene Expression in Neural Stem Cells Through Polymeric Nanoparticles.

Therapy based on stem cells utilizes these cells in neurodegeneration, brain/spinal cord injury, and much recently in repairing of severe heart diseases. Owning to their stemness, these cells are the potential source of progenitors that can offer a therapeutic remedy to a variety of diseases and/or disorders. The ability of these cells to regenerate and differentiate into specified phenotypes has great utility in tissue regeneration applications. This chapter provides a detailed account for isolation of neural stem cells from the mice embryo. Furthermore, the fabrication of chitosan-tripolyphosphate/hyaluronic acid-based nanoparticles and evaluating their efficiency in inducing transfection in the isolated neural stem cells as an approach for the treatment of neurodegenerative disorders.

Protocol for Determining the Induction of Human Embryonic Stem Cells into Myogenic Lineage Using Electrospun Nanofibers.

An efficient method for the development of myogenic differentiation using the stem cells can be beneficial in patients with severely compromised mobility, muscular damage, or degenerative diseases. The stem cells can prove to be excellent clinical source of myogenic progenitor cells due to their ability of self-proliferation, renewal, and differentiation into a specific phenotype. They represent an essential component of tissue engineering along with other factors (e.g., 3D scaffolds, ECM mimicking environment, and growth factors). In this chapter, we describe the experimental protocols for isolation of the embryonic stem cells, their proliferation on nanofiber scaffolds, and finally their differentiation into myogenic cells. Furthermore, this chapter elaborates experimental methods to assess the myogenic fate of embryonic stem cells on the nanofiber scaffolds.