Identification of common candidate genes and pathways for Spina Bifida and Wilm's Tumor using an integrative bioinformatics analysis.

Spina Bifida (SB) and Wilm's Tumor (WT) are conditions, both associated with children. Several studies have shown that WT later develops in SB patients, which led us to elucidate common key genes and linked pathways of both conditions, aimed at their concurrent therapeutic management. For this, integrated bioinformatics analysis was employed. A comprehensive manual curation of genes identified 133 and 139 genes associated with SB and WT, respectively, which were used to construct a single protein-protein interaction (PPI) network. Topological parameters analysis of the network showed its scale-free and hierarchical nature. Centrality-based analysis of the network identified 116 hubs, of which, 6 were called the key genes attributed to being common between SB and WT besides being the hubs. Gene enrichment analysis of the 5 most essential modules, identified important biological processes and pathways possibly linking SB to WT. Additionally, miRNA-key gene-transcription factor (TF) regulatory network elucidated a few important miRNAs and TFs that regulate our key genes. In closing, we put forward TP53, DICER1, NCAM1, PAX3, PTCH1, MTHFR; hsa-mir-107, hsa-mir-137, hsa-mir-122, hsa-let-7d; and YY1, SOX4, MYC, STAT3; key genes, miRNAs and TFs, respectively, as the key regulators. Further, MD simulation studies of wild and Glu429Ala forms of MTHFR proteins showed that there is a slight change in MTHFR protein structure due to Glu429Ala polymorphism. We anticipate that the interplay of these three entities will be an interesting area of research to explore the regulatory mechanism of SB and WT and may serve as candidate target molecules to diagnose, monitor, and treat SB and WT, parallelly.Communicated by Ramaswamy H. Sarma.

Outcome of Neuromuscular Electrodiagnostic Testing in Children.

This study aimed to describe the spectrum of the results of neuromuscular electrodiagnostic investigations performed by age and pathology in paediatric patients. This retrospective observational study took place from January 2019 to December 2021. Electrodiagnostic records of paediatric patients (birth to 15 years) were referred to the electromyography (EMG) laboratory for EMG and nerve conduction study (NCS). Electrodiagnostic and demographic data were collected. Among these, 151 (56%) were males and 118 (44%) were females. Forty-four percent tests came out to be normal and 151(56%) tests revealed electrophysiological abnormality. Peripheral nerve neuropathy (n = 39, 14.5%), polyneuropathy (n = 35, 13%) and myopathy (n = 28, 10%) were the most common electrophysiological diagnoses. The electrodiagnostic test helps in differentiating the aetiologies, thus guiding the subsequent work-up logically in the paediatric population. Key Words: Brachial plexus neuropathies, Electromyography, Duchene muscular dystrophy, Neuromuscular junction disorders, Nerve conduction studies, Acute inflammatory demyelinating polyneuropathy, Rehabilitation, Electromyography, Hereditary sensory motor polyneuropathy.

The United Kingdom National Health Service through the lens of an overseas clinical observer: A reflective account based on personal experience.

This article provides insight into an aspect of the National Health Service (NHS) of the United Kingdom (UK) through the perspective of an overseas clinical observer. The NHS is a government-funded medical and health care service in the UK and has developed over the years providing a high standard of medical care and professionalism. The Clinical Observership Programme has been designed primarily for International Medical Graduates to experience how the NHS functions and the kind of services it offers. This article explores the overseas observer's experience in a surgical unit of the NHS including the organisational structure, delivery of care and challenges of the system. This article also highlights the observer's perspective of surgical care in the NHS compared to surgical care in low-resource income countries. The Clinical Observership is useful in learning about UK medical practice and gaining experience in a different cultural and professional environment. This is a reflective first-hand account based on personal experience.

Pharmacological Activities of Rhododendron afghanicum; an Endemic Species of Khyber Pakhtunkhwa, Pakistan.

Majority of different kinds of metabolites having therapeutic characteristics are thought to be stored in medicinal plants. So, the present study was aimed to explore the crude extract of leaves and stem of R. afghanicum for phytochemical screening and various pharmacological activities. Toxicological studies at 100 mg/kg showed 60 % mortality where its safe dose level was 90 mg/kg. Phytochemical screening revealed the presence of alkaloids, glycosides, flavonoids and tannins in both extracts. Bacterial strains were susceptible to (RLEt) and (RLM) crude extracts except Staphylococcus aureus. RSM showed maximum anti-inflammatory activity (20.16 %) followed by RSEt (20.14 %) where lowest activity was displayed by RLEt (18.46 %). Phytotoxic activity showed a substantial dose-dependent phyto-inhibition of Lemna minor. An outstanding cytotoxic potential was displayed with LD50 values of 9.46 and 13.03 µg/ml in both stem extracts. RLEt demonstrated a dose-dependent pain relief at 30, 60 and 90 mg/kg which was 31 %, 40 % and 52 % respectively. A considerable spasmolytic action was observed by the shrinkage of jejunum muscle in albino mice. RLEt at 1000 ppm showed (17 mm) and RLM at 1000 ppm showed (16 mm) zone of inhibition against Aspergillus niger. These findings support and corroborate the traditional applications of R. afghanicum for treating digestive, analgesic and inflammatory ailments.

Mucoadhesive, Fluconazole-Loaded Nanogels Complexed with Sulfhydryl-ß-cyclodextrin for Oral Thrush Treatment.

The objective of this study was to generate fluconazole-loaded mucoadhesive nanogels to address the problem of hydrophobicity of fluconazole (FL). An inclusion complex was formulated with sulfhydryl-ß-CD (SH-ß-CD) followed by nanogels formation by a Schiff base reaction of carbopol 940 (CA-940) and gelatin (GE). For characterization, PXRD, FT-IR analysis, drug content, and phase solubility studies were performed. Similarly, nanogels were assessed for particle size, zeta potential, organoleptic, and spreadability studies. Moreover, drug contents, rheological, in vitro drug permeation, release kinetics, toxicity, and stability studies of nanogels were performed. Furthermore, mucoadhesive characteristics over the buccal mucosal membrane of the goat were evaluated. The nanogels formulated with a higher amount of CA-940 and subsequently loaded with the inclusion complexes of FL showed promising results. PXRD and FT-IR analysis confirmed the physical complexes by displaying a reduction in the intensity of peaks of FL. The average particle size of nanogels was in the range of 257 to 361 nm. The highest drug content of 88% was encapsulated within the FL-SH-ß-CD complex. All formulations at 0.5-1% concentration displayed no toxicity to the Caco-2 cell lines. Nanogels loaded with FL-SH-ß-CD complexes showed 18-fold improved mucoadhesion on the buccal mucous membrane of the goat when compared to simple nanogels. The in vitro permeation study exhibited significantly enhanced permeation and first-order concentration-dependent drug release was observed. On the bases of these findings, we can conclude that a mucoadhesive nanogel-based drug delivery system can be an ideal therapy for candidiasis.

Impact of Some Toxic Metals on Important ABC Transporters in Soybean (Glycine max L.).

In plants, ATP-binding cassette (ABC) transporters facilitate the movement of substrates across membranes using ATP for growth, development, and defense. Soils contaminated with toxic metals such as cadmium (Cd) and mercury (Hg) might adversely affect the metabolism of plants and humans. In this study, a phylogenetic relationship among soybeans' (Glycine max) ATP binding cassette (GmABCs) and other plant ABCs was analyzed using sequence information, gene structure, chromosomal distribution, and conserved motif-domain. The ontology of GmABCs indicated their active involvement in trans-membrane transport and ATPase activity. Thirty-day-old soybean plants were exposed to 100 µM CdCl2 and 100 µM HgCl2 for 10 days. Physiological and biochemical traits were altered under stress conditions. Compared to Control, GmABC transporter genes were differentially expressed in response to Cd and Hg. The qRT-PCR data showed upregulation of seven ABC transporter genes in response to Cd stress and three were downregulated. On the other hand, Hg stress upregulated four GmABC genes and downregulated six. It could be concluded that most of the ABCB and ABCG subfamily members were actively involved in heavy metal responses. Real-time expression studies suggest the function of specific ABC transporters in Cd and Hg stress response and are helpful in future research to develop stress-tolerant varieties of soybean.

In Silico and In Vitro Studies of 4-Hydroxycoumarin-Based Heterocyclic Enamines as Potential Anti-Tumor Agents.

The present study reports the one-step synthesis of several 3-formyl-4-hydroxycouramin-derived enamines (4a-4i) in good yields (65-94%). The characterization of the synthesized compounds was carried out via advanced analytical and spectroscopic techniques, such as melting point, electron impact mass spectrometry (EI-MS), 1H-NMR, 13C-NMR, elemental analysis, FTIR, and UV-Visible spectroscopy. The reaction conditions were optimized, and the maximum yield was obtained at 3-4 h of reflux of the reactants, using 2-butanol as a solvent. The potato disc tumor assay was used to assess Agrobacterium tumefaciens-induced tumors to evaluate the anti-tumor activities of compounds (4a-4i), using Vinblastine as a standard drug. The compound 4g showed the lowest IC50 value (1.12 ± 0.2), which is even better than standard Vinblastine (IC50 7.5 ± 0.6). For further insight into their drug actions, an in silico docking of the compounds was also carried out against the CDK-8 protein. The binding energy values of compounds were found to agree with the experimental results. The compounds 4g and 4h showed the best affinities toward protein, with a binding energy value of -6.8 kcal/mol.

Short-Term Outcomes and Quality-of-Life Assessment Following Rives-Stoppa and Transversus Abdominis Release Procedures of Open Ventral Hernia Repair.

INTRODUCTION: Ventral hernia is one of the common surgical conditions that can significantly impact a patient's quality of life (QoL). Open ventral hernia repair using the Rives-Stoppa (RS) and Transversus Abdominis Release (TAR) procedures has gained recognition for its effectiveness in achieving hernia repair and reducing the risk of further recurrence. However, limited research has been performed to explore the short-term outcomes and QoL assessment following these two surgical techniques. The aim of this study was to know the result after RS and TAR methods of hernia repair in terms of short-term recurrences, pain, postoperative complications, and QoL. METHODS: This was a prospective, interventional study, which included 30 patients fulfilling the inclusion criteria. The study group was subjected to posterior component separation (PCS)-TAR and RS repair as per surgical indication (RS if defect size 4-10cm; PCS-TAR if defect size >10cm and

PEGylated Graphene Oxide as a Nanodrug Delivery Vehicle for Podophyllotoxin (GO/PEG/PTOX) and In Vitro α-Amylase/α-Glucosidase Inhibition Activities.

This study aims to develop a nanodrug delivery system containing podophyllotoxin (PTOX), a known anticancer drug, loaded on graphene oxide (GO). The system's ability to inhibit α-amylase and α-glucosidase enzymes was also investigated. PTOX was isolated from Podophyllum hexandrum roots with a yield of 2.3%. GO, prepared by Hummer's method, was converted into GO-COOH and surface-mobilized using polyethylene glycol (PEG) (1:1) in an aqueous medium to obtain GO-PEG. PTOX was loaded on GO-PEG in a facile manner with a 25% loading ratio. All the samples were characterized using FT-IR spectroscopy, UV/visible spectroscopy, and scanning electron microscopy (SEM). In FT-IR spectral data, GO-PEG-PTOX exhibited a reduction in acidic functionalities and there was an appearance of the ester linkage of PTOX with GO. The UV/visible measurements suggested an increase of absorbance in 290-350 nm regions for GO-PEG, suggesting the successful drug loading on its surface (25%). GO-PEG-PTOX exhibited a rough, aggregated, and scattered type of pattern in SEM with distinct edges and binding of PTOX on its surface. GO-PEG-PTOX remained potent in inhibiting both α-amylase and α-glucosidase with IC50 values of 7 and 5 mg/mL, closer to the IC50 of pure PTOX (5 and 4.5 mg/mL), respectively. Owing to the 25% loading ratio and 50% release within 48 h, our results are much more promising. Additionally, the molecular docking studies confirmed four types of interactions between the active centers of enzymes and PTOX, thus supporting the experimental results. In conclusion, the PTOX-loaded GO nanocomposites are promising α-amylase- and α-glucosidase-inhibitory agents when applied in vitro and have been reported for the first time.

National TB elimination programme - What has changed.

BACKGROUND: Before the start of Coronavirus (COVID-19) pandemic, TB was the leading cause of death due to a single infectious agent, ranking well above HIV/AIDS. Almost one-fourth of the world's population is infected with M. tuberculosis. TB is curable and preventable. About 85% of people who develop TB can be successfully treated with drug regimens of 6 months. Universal health coverage (UHC) is necessary to ensure that all those with the disease can access these treatments. Research breakthroughs (e.g., newer rapid diagnostic techniques, drugs, newer vaccine) are needed to rapidly reduce the number of new cases each year (TB incidence) worldwide. OBJECTIVE: Changes in the National TB Elimination Programme since its inception. CONTENT: The Government of India launched the "National TB Programme" in 1962 as District TB Centre model involved with BCG vaccination and TB treatment to fight tuberculosis, a major public health problem. The tuberculosis control programme has come a long way since then and has undergone major changes over the past few years The Ministry of Health and Family Welfare has developed the "National Strategic Plan" for Tuberculosis Elimination (2017-25) which encapsulates the bold and innovative steps required to eliminate TB in India by 2025, five years ahead of the global targets. By 2020 it was clear that the NSP- 2017-25 will not be able to meet these objectives, so another new NSP India 2025 had been launched in 2020. India has been actively involved in TB control activities for more than 50 years now. TB still continues to be a severe health problem in India. The country is now better prepared to tackle TB than before. It now has advanced and effective interventions and technologies for diagnosis, treatment and care of TB cases.

Seasonal Investigation of Anaplasma marginale Infection in Pakistani Cattle Reveals Hematological and Biochemical Changes, Multiple Associated Risk Factors and msp5 Gene Conservation.

Bovine anaplasmosis is a tick-borne disease caused by an obligate intercellular Gram-negative bacterium named Anaplasma (A.) marginale. In this study, we report the seasonal prevalence, potentially associated risk factors and phylogeny of A. marginale in cattle of three different breeds from Multan District, Southern Punjab, Pakistan. A total of 1020 blood samples (crossbred, n = 340; Holstein Friesian, n = 340; and Sahiwal breed, n = 340) from apparently healthy cattle were collected on a seasonal basis from March 2020 to April 2021. Based on PCR amplification of the msp5 partial sequence, overall, the A. marginale prevalence rate was estimated at 11.1% (113/1020) of the analyzed cattle samples. According to seasons, the highest prevalence rate was observed in autumn (16.5%), followed by winter (10.6%) and summer (9.8%), and the lowest was recorded in the spring (7.5%). The crossbred and Sahiwal cattle were the most susceptible to A. marginale infection, followed by Holstein Friesian cattle (7.9%). Analysis of epidemiological factors revealed that cattle reared on farms where dairy animals have tick loads, dogs coinhabit with cattle and dogs have tick loads have a higher risk of being infected with A. marginale. In addition, it was observed that white blood cell, lymphocyte (%), monocyte (%), hematocrit, mean corpuscular hemoglobin and mean corpuscular hemoglobin concentrations were significantly disturbed in A. marginale-positive cattle compared with non-infested cattle. Genetic analysis of nucleotide sequences and a phylogenetic study based on msp5 partial sequencing demonstrated that this gene appears to be highly conserved among our isolates and those infecting apparently healthy cattle from geographically diverse worldwide regions. The presented data are crucial for estimating the risk of bovine anaplasmosis in order to develop integrated control policies against bovine anaplasmosis and other tick-borne diseases infecting cattle in the country.

Recent advances in yeast genome evolution with stress tolerance for green biological manufacturing.

Green biological manufacturing is a revolutionary industrial model utilizing yeast as a significant microbial cell factory to produce biofuels and other biochemicals. However, biotransformation efficiency is often limited owing to several stress factors resulting from environmental changes or metabolic imbalance, leading to the slow growth of cells, compromised yield, and enhanced energy consumption. These factors make biological manufacturing competitively less economical. In this regard, minimizing the stress impact on microbial cell factories and strong robust performance have been an interesting area of interest in the last few decades. In this review, we focused on revealing the stress factors and their associated mechanisms for yeast in biological manufacturing. To improve yeast tolerance, rational and irrational strategies were introduced, and the molecular basis of genome evolution in yeast was also summarized. Furthermore, strategies of genome-directed evolution such as homology directed repair and nonhomologous end-joining, and the synthetic chromosome recombination and modification by LoxP-mediated evolution and their association with stress tolerance was highlighted. We hope that genome evolution provides new insights for solving the limitations of the natural phenotypes of microorganisms in industrial fermentation for the production of valuable compounds.

Pharmacognostic standardization of Rhododendron afghanicum through scanning electron microscopy and analytical techniques.

The aim of this study was to assess and compare the pharmacognostic and microscopic features of the selected parts of Rhododendron afghanicum Aitch. & Hemsl. It is a perennial and shrub. Anatomy of stem and leaves depicted dicot structure. It will provide knowledge about standardization, authentication, and adulteration with its co-species. The macroscopic examination reveals that fresh leaves are green, pleasant odor with a bitter taste. The leaf is oblong-elliptic in shape and sub-acute at apex; rounded at the base with entire margin. Stem is irregular and the outer surface is light brown, rough with fissures and ridges. Microscopic examination indicated the presence of xylem, phloem, peltate trichome, epidermal cells, collenchymas cells, paracytic stomata, and reticulate vessels. Stem microscopy reveals epidermis, hypodermis, cortex, sclerenchymatous sheath, phloem, xylem, and pith. It will be helpful in identification and quality control. Micro-morphological features were observed through SEM. EDX spectroscopy were carried out and revealed the presence of calcium, silicon, and potassium. Phytochemical analysis revealed the presence of tannins, saponins, phenols, protein, flavonoids, glycosides, and alkaloids. Ethyl acetate for leaf and stem demonstrate the highest extractive values (18% and 13%), respectively. XRD peaks appeared at 30.21, 28.73, 205.73, 200.73, 380.07, 390.24, 490.11, and 450.33ο . This will be helpful to identify the ownership of herbal drugs by the diffraction peaks through crystal structures and atomic spacing. These parameters are crucial for drug identification, standardization, authentication, and drug designing. These studies also provided knowledge regarding therapeutic and nutraceutical importance of this plant. RESEARCH HIGHLIGHTS: The current pharmacognostic studies carried out on Rhododendron afghanicum can serve as a basis for compiling keys for finding the taxonomic identification and authentication of the said species by morphological, anatomical, and physicochemical features.

Evaluation of Pakistani wheat germplasm for leaf rust resistance at various locations.

341 entries comprising of 250 genotypes/lines and 91 gene differentials were tested for leaf rust (Puccinia triticina Erik) in different ecological zones of Punjab during 2016-17 and 2017-18. Each entry was planted in a single 1 m long row and Morocco was used as a spreader. Data on leaf rust severity was recorded once in 3rd week of March during both study years at all locations by following Modified Cobb Scale while the data was recorded three times on 2nd, 22nd and 29th March during 2018 at Faisalabad location to study rust development pattern. The disease severity ranged from 0-100S during 2016-17 and from 0-80S during 2017-18. The genotype HYT 60-5 and the genes Lr-19, Lr-26 and Lr 27+31 showed no disease symptoms at any location during both the study years. These genes can be used for future breeding material development. Area under disease progressive curve (AUDPC), calculated on the basis of periodical readings from Faisalabad, ranged from 0-550 and the susceptible check Morocco has AUDPC value of 600. 120 entries including HYT 60-5 have disease progression 0, which showed that there may be a major gene based resistance in these entries. Area under disease progressive curve/Day (AUDPC/DAY) was calculated for the rest of 130 genotypes to have an understanding of the disease progression pattern and out of which 43 entries have AUDPC/Day value ranging from 1-2 and 28 entries have AUDPC/Day value ranging from 2-3 which revealed that these entries are very useful for use in breeding for durable rust resistance and can be utilized as a parent in back cross and top cross breeding schemes. Material with AUDPC value less than 10 is the best source of resistance against the leaf rust. Varieties/advanced lines, Ujala-16, V-14154, and V-14124 have shown slow rust development and are very good sources of resistance. Similarly, HYT 60-5 has proven an excellent source of resistance. The advance line V-14154 has been approved as a commercial cultivar by the name "Akbar-19".

Measuring Readiness for Self-Directed Learning in Medical Undergraduates.

Purpose: To measure the readiness for self-directed learning in medical students for the evaluation of self-directed learning in the study population. Materials and Methods: The survey was conducted in undergraduate students. The instrument used was Fisher's readiness scale comprising of self-management, desire for learning, and self-control domains. The data were analyzed by Mann-Whitney U-test and bivariate and partial correlations. The results were compared with the reported ones. Results: Total students surveyed were 300. Of these, 96 responded - 73 (76%) of preclinical and 23 (24%) of clinical classes. The mean readiness score was 124. The mean domains' scores for self-management, desire for learning, and self-control were 38, 38, and 48, respectively. The preclinical group had a mean score of 122 for readiness,  37 for self-management and desire for learning each, and 48 for self-control. The clinical group's scores were 129, 40, and 49, respectively. Preclinical and clinical groups differed significantly in self-management domain (P = 0.03). The difference was not significant in desire for learning (P = 0.08), self-control domains (P = 0.40) and readiness score (P = 0.12). The domains of self-control and desire for learning had a positive correlation if self-management was controlled, and self-control and self-management had a positive correlation if desire for learning was controlled (P < 0.05). Conclusion: The measurement of readiness for self-directed learning helps in knowing the true value of self-directed learning in a particular setting. Relatively lower scores in our study mean self-directed learning alone cannot be relied upon to achieve optimum students' learning. There is also a need for implementing strategies that will help students in improving their readiness for independent learning.

Global Gene Expression and Docking Profiling of COVID-19 Infection.

Coronavirus is an enclosed positive-sense RNA virus with club-like spikes protruding from its surface that causes acute respiratory infections in humans. Because it is considered a member of the complex pathogen group, it has been found to infect different host species and cause a variety of diseases. So far, it has been discovered that it may affect the immune, infection, and inflammatory systems, leading to the hypothesis that the immune and inflammatory systems (signaling pathways and components) fail to control infection, opening the door to look for potential targets primarily in these systems. The study's main purpose is to identify highly overexpressed genes and their functional implications as a result of COVID-19 infection, as well as to investigate probable infections, inflammation, and immune systems to better understand the impact of coronavirus infection. We explored the genes and pathways mostly linked with infection, inflammation, and the immune systems using the datasets available for COVID-19 infection gene expression compendium. NFKBIA, FN1, FAP, KANK4, COMP, FAM101B, COL1A2, ANKRD1, TAGLN, SPARC, ADAM19, OLFM4, CXCL10/11, OASL, FOS, APOBEC3A, IFI44L, IFI27, IFIT1, RSAD2, NDUFS1, SRSF6, HECTD1, CBX3, and DDX17 are among the genes that may be impacted by infection, according to our findings. The functional changes are mainly associated with these pathways TNF, cytokine, NF-kB, TLR, TCR, BCR, Foxo, and TGF signaling pathways are among them and there are additional pathways such as hippo signaling, apoptosis, estrogen signaling, regulating pluropotency of stem cells, ErbB, Wnt, p53, cAMP, MAPK, PI3K-AKT, oxidative phosphorylation, protein processing in endoplasmic reticulum, prolactin signaling, adipocytokine, neurotrophine signaling, and longevity regulating pathways. Moreover, we have also explored the potential herbal drug (apigenin, quercetin, and resveratrol) targets for the top-rated genes based on the overall analysis where we observe that quercetin and resveratrol as most effective.

Emerging role of oncolytic viruses and stem cells in gene therapy: Should they be integrated?

Recombinant virus-based transgene therapy has shown promising results in solid tumors. Oncolytic virotherapy is a research hotspot because of its additional immunostimulatory effects. However, metastatic malignancies require systemic virotherapy, which necessitates the use of safe and effective vehicles for drug delivery. Mesenchymal stem cells (MSCs) are good carriers because of their tumor-tropic and immune-evasive capabilities. We collated published results from pre-clinical and clinical trials to support the use of MSCs as Trojan horses for the systemic administration of recombinant viruses, with a focus on glioblastoma. The generation of modified MSCs harboring recombinant viruses could expedite bench-to-bedside transformation.

Efficacy of Magnesium Sulfate in Addition to Melatonin Therapy in Neonates With Hypoxic-Ischemic Encephalopathy.

BACKGROUND: One of the most important causes of neonatal deaths in developing nations is birth asphyxia. Though the probability of a complete recovery is very low, hypoxic-ischemic encephalopathy (HIE) associated with asphyxia can be managed with multiple treatment options. The study evaluated the efficacy of the addition of magnesium sulfate (MgSO4) to melatonin therapy in neonates with HIE. METHODOLOGY: A prospective, observational study was conducted in the department of neonatal intensive care, Pakistan Institute of Medical Sciences Hospital, Islamabad, Pakistan from October 2020 to March 2021. All neonates with an Appearance, Pulse, Grimace, Activity, and Respiration (APGAR) score of less than five at five minutes, umbilical blood pH of less than 7.0, and moderate neonatal encephalopathy as detected on the modified Sarnat score which is a clinical tool used for the assessment of the severity of HIE were included in the study. Neonates with congenital abnormalities, intrauterine growth retardation, neonatal sepsis, and infants born to mothers with diabetes mellitus type 2 were excluded from the study. The infants were randomly assigned to either of the groups, i.e., i) group 1 included neonates who were administered at least three doses of magnesium sulfate (MgSO4) infusion in addition to melatonin, or ii) group 2 included neonates who were administered melatonin only. Blood samples of all neonates were evaluated and compared between the two groups. RESULTS: A total of 90 neonates with HIE were included in the study. There was a predominance of female neonates. The mean ages of babies in group 1 and group 2 were 37.2 ± 0.43 and 37.3 ± 0.59 weeks, respectively. The mean weight on the term was 2.88 ± 0.11 and 2.89 ± 0.10, respectively. The Apgar score at 5 mins in group 1 was 1.73 ± 0.81 while in group 2, 1.82 ± 0.94. It was found that there was a more significant improvement in pH after 3 days and after one week of treatment in group 1 as compared to group 2. The mean pH in babies after three days of intervention was 7.23 ± 0.03 in group 1 which was significantly better than group 2 (p<0.0001). After seven days, the mean normalized to 7.39 ± 0.04 in group 1 (p < 0.0001). It was found that in patients in group 1, the mortality was lower than in group 2 (p < 0.0001). CONCLUSION: HIE patients who were administered melatonin in combination with magnesium sulfate yielded better patient outcomes. Thus, it was concluded that the use of magnesium sulfate as dual therapy with melatonin improved patient outcomes for HIE. However, it is recommended that similar studies are conducted with a wider range of parameters, such as duration of hospital stay and assessment of the neurological outcomes of the patients.

Molecular study on the role of vacuolar transporters in glycyrrhetinic acid production in engineered Saccharomyces cerevisiae.

Glycyrrhetinic acid (GA) is one of the major bioactive components of the leguminous plant, Glycyrrhiza spp. (Chinese licorice). Owing to GA's complicated chemical structure, its production by chemical synthesis is challenging and requires other efficient strategies such as microbial synthesis. Earlier investigations employed numerous approaches to improve GA yield by refining the synthetic pathway and improving the metabolic flux. Nevertheless, the metabolic role of transporters in GA biosynthesis in microbial cell factories has not been studied so far. In this study, we investigated the role of yeast ATP binding cassette (ABC) vacuolar transporters in GA production. Molecular docking of GA and its precursors, ß-Amyrin and 11-oxo-ß-amyrin, was performed with five vacuolar ABC transporters (Bpt1p, Vmr1p, Ybt1p, Ycf1p and Nft1p). Based on docking scores, two top scoring transporters were selected (Bpt1p and Vmr1p) to investigate transporters' functions on GA production via overexpression and knockout experiments in one GA-producing yeast strain (GA166). Results revealed that GA and its precursors exhibited the highest predicted binding affinity towards BPT1 (ΔG = -10.9, -10.6, -10.9 kcal/mol for GA, ß-amyrin and 11-oxo-ß-amyrin, respectively). Experimental results showed that the overexpression of BPT1 and VMR1 restored the intracellular as well as extracellular GA production level under limited nutritional conditions, whereas knockout of BPT1 resulted in a total loss of GA production. These results suggest that the activity of BPT1 is required for GA production in engineered Saccharomyces cerevisiae.