Search

The os trigonum, an irregular, accessory ossicle at the posterior lateral process of the talus, is known to be one of the main causes of posterior ankle impingement. It is unclear if an arthroscopic technique offers a superior outcome over an open technique when surgical excision is indicated. The purpose of this study is to better understand the outcomes and complications after os trigonum excision using an open versus arthroscopic technique. Using PRISMA guidelines, a systematic review of studies published in PubMed, Cochrane Reviews, and CINAHL databases through November 2021 was conducted to identify articles that evaluated postoperative outcomes and complications following os trigonum excision. Studies involving concurrent procedures, arthroscopic approaches varying from the technique described by Van Dijk et. al (2000), and case studies (where n ≤ 2) were excluded 1996 unique articles were identified. 18 studies were included representing 242 patients undergoing the arthroscopic technique and 217 undergoing the open technique. The arthroscopic group returned to activity sooner than the open group (9.86 weeks versus 13.074 weeks, respectively; p-value

The aim of the present work was to prepare microgel nanocomposites based on silver and magnetite to apply as adsorbents and heterogeneous catalysts for removal of methylene blue (MB) cationic dye from aqueous solution. For this, 2‐acrylamido‐2‐methylpropane sulfonic acid (AMPS) and acrylamide (AAm) monomers were used to prepare AMPS/AAm microgel based on the emulsion technique. Ag and Fe3O4 nanoparticles were embedded into the AMPS/AAm microgel using the in situ technique. Their particle sizes, surface charges, crystalline lattice structure, morphology, magnetic properties and thermal stability were investigated. The AMPS/AAm hydrogel nanocomposites were used as an adsorbent to remove MB dye. The AMPS/AAm microgel nanocomposites were tested as catalysts to reduce MB and degrade its chemical structure with heterogeneous Fenton oxidation using Ag and Fe3O4 nanocomposites, respectively. This study presents promising data as the prepared materials used as adsorbents and catalysts show competitive features compared with the data presented in the literature. © 2019 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Epsilon toxin (ETX) is an exotoxin produced by Clostridium perfringens type D that induces enterotoxaemia or necrotic intestinal infection in small ruminants and bovine. Immunization is an essential element in preventing the spread of infectious diseases. In recent literature, nanocarriers have exhibited the capacity to deliver protection, stability, and regulated distribution properties to protein-based antigens. Furthermore, egg albumin is a highly adaptable protein nanocarrier in vaccine delivery systems due to its biocompatible, biodegradable, non-toxic, and non-immune-modulating properties. In this study, we assessed the efficacy, safety, immunogenicity, and dose-effect relationships of the nanoparticle-advanced toxoid vaccine (G1) in contrast to the commercially available vaccine (ETV) (G2). Two different vaccines (1 ml) were inoculated in experimental animals (rabbits) on days 1, 7, 14, 21, and 28. The geometric mean titers (GMT) of Groups 2 and 3 were recorded on the respective day of inoculation. The findings reveal that the GMT of group 2 was significantly higher than group 3. The use of nanoparticles to detain toxins demonstrated enhanced immune protection against the harmful effects caused by the toxins. This work is anticipated to explore new opportunities in developing improved vaccinations using nanoparticles to combat the pathogenicity/ virulence factors that present potential risks to livestock., Competing Interests: Declarations. Competing interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

Background: New strains of SARS-CoV-2 are continually emerging worldwide. Recently, WHO warned of a severe new wave in Europe. Current vaccines cannot fully prevent reinfection in vaccinated individuals., Aim: Given this issue, recent research focuses on new antiviral candidates with high efficacy and minimal side effects., Objectives: Screen natural compounds as inhibitors of Mpro SARS-CoV-2 protein using molecular dynamics., Methods: In this study, we have screened the potential of plant-based natural anti-viral compounds. A library of the 579 compounds was generated using currently available literature and online databases. All these compounds were screened based on their binding affinities as predicted by molecular docking analysis and compounds having binding affinity values ≤ -10 Kcal/mol were considered for analysis. Furthermore, from physicochemical assessment, drug-likeness initially nine compounds were identified as the antiviral targets for the selected viral proteins. After ADMET analysis and simulations, the compound 9064 with the lowest RMSD, Coul-SR interaction energy (-71.53 kJ/mol), and LJ-SR energy (-95.32 kJ/mol) was selected as the most stable drug candidate against COVID-19 main protease Mpro., Results: The ΔG value, calculated using MMGBSA also revealed strong binding of the compound with Mpro. The selected antiviral compound 9064 is an antioxidant flavonoid (Catechin or Cianidanol), which was previously known to have significant immunomodulatory, anti-inflammatory, and antioxidant properties., Conclusion: Considering the limitations of currently available vaccines, our study may provide new insight into potential drugs that may prevent SARS-CoV-2 infection in humans., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Objectives: To assess the feasibility of using a faecal occult blood test (FOBT) kit as a screening tool for periodontal disease. Additionally, the study aimed to evaluate the test's sensitivity and specificity while exploring the influence of factors like age, sex and the number of teeth on its performance., Methods: A prospective observational case-control study involving 150 participants divided into three groups on the basis of the basic periodontal examination index: healthy (n = 21), gingivitis (n = 52) and periodontitis (n = 77). Salivary occult blood tests were conducted using the Onsite FOB-Hi Rapid Test. Descriptive and analytical statistical analyses were performed using Fisher's exact test and chi-square tests., Results: The FOBT kit demonstrated a sensitivity of 91% and a specificity of 100%. The number of teeth significantly influenced test results (p value = 0.021, chi-square). Age (p value = 0.09, chi-square) and sex (p value = 0.213, chi-square) did not significantly impact the test's performance., Conclusion: The FOBT kit demonstrates high sensitivity (91.3%) and specificity (100%) as a screening tool for periodontal disease. Its affordability and global availability make it suitable for detecting periodontal disease across different age groups, sexes and numbers of teeth., (© 2024 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background and Aim: Avian influenza is a global threat to avian species, particularly in developing countries. Recombinant vaccines, including virus-like particles (VLPs), are promising strategies for preventing the spread of the disease. VLPs produced through the self-assembly of viral structural proteins without genomic material mimic native virions and are promising platforms for new vaccines. VLPs have been shown to elicit protective antibodies and are effective and safe vaccines against influenza. This study aimed to optimize the protocol for the production and characterization of H9N2 VLPs and their evaluation as a vaccine in broiler birds., Materials and Methods: Low-pathogenic influenza virus (LPAI) H9N2 was isolated and characterized through whole-genome sequencing, and a VLP-based vaccine for LPAI H9N2 was prepared using a baculovirus expression system. Codon-optimized hemagglutinin (HA) , neuraminidase (NA) , and M1 were successfully cloned in pFastbac1 and expressed in SF9 cells. Proteins were characterized using sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE), western blotting, and electron microscopy after purification. Semi-purified proteins were tested as a vaccine in broiler chickens challenged with LPAI H9N2., Results: Recombinant Bacmid DNA from positive clones was extracted and confirmed using a polymerase chain reaction. The transfection showed cytopathic effects, and the proteins were confirmed through western blotting and SDS-PAGE, which showed the sizes of HA = 62-64 KD, NA = 52 KD, and M1 = 25 KD. The shape and morphology were confirmed through transmission electron microscopy which revealed 100-150 nm size particles. As a result, the semi-purified VLPs (HA assay: 256) were tested as a vaccine for specific-pathogen free broiler birds; administered through subcutaneous and intranasal routes. The birds were challenged on the 28 th day after vaccination with the H9N2 strain, and the birds showed significant cross-reactivity with the H9N2 strain., Conclusion: The semi-purified VLP-based vaccine induced a significant immune response in vivo . This vaccine formulation has the potential to control avian influenza outbreaks in Pakistan's poultry industry., Competing Interests: The authors declare that they have no competing interests., (Copyright: © Luqman, et al.)

Objectives: To evaluate the efficacy of platelet-rich fibrin (PRF) as an adjunct to scaling and root planing (ScRp) for healing shallow periodontal pockets., Methods: Twelve patients with periodontitis were enrolled in this split-mouth, randomized clinical trial. A total of 24 shallow periodontal pockets (4-6 mm) were treated by either ScRp alone (control) or PRF (test). Clinical attachment loss (CAL), probing pocket depth (PPD), bleeding on probing (BOP), and plaque index (PLI), as well as platelet-derived growth factor-BB (PDGF-BB) by enzyme-linked immunosorbent assay (ELISA) in gingival crevicular fluid (GCF) were measured at baseline and at 1- and 3-month follow-up visits., Results: At 1- and 3-month follow-up visits, greater CAL gains (2.6 ± 0.25 mm and 3.26 ± 0.31 mm, respectively) and PPD reductions (2.58 ± 0.38 and 3.31 ± 0.39 mm, respectively) were observed in the test group compared to those in controls (CAL gain of 1.01 ± 0.49 mm and 1.43 ± 0.48 mm; PPD reduction of 1.1 ± 0.55 and 1.37 ± 0.49 mm, respectively). In addition, the increase in PDGF-BB in GCF in the test group (724.5 ± 186.09 pg/μl and 1957.5 ± 472.9 pg/μl) was significantly greater than that in controls (109.3 ± 24.07 and 614.64 ± 209.3 pg/μl) at 1- and 3-month follow-up visits, respectively., Conclusions: The noninvasive use of PRF as an adjunct to ScRp successfully improved clinical periodontal parameters and might contribute to increased PDGF-BB in GCF., Competing Interests: Declaration of competing interest All the authors certify that they have no conflict of interest to disclose in relation to the subject matter or materials discussed in the present study., (Copyright © 2024 Japanese Association for Oral Biology. Published by Elsevier B.V. All rights reserved.)

Peste des petits ruminants (PPR) is an extremely transmissible viral disease caused by the PPR virus that impacts domestic small ruminants, namely sheep and goats. This study aimed to employ a methodical approach to evaluate the regional occurrence of PPR in small ruminants in Pakistan and the contributing factors that influence its prevalence. A thorough search was performed in various databases to identify published research articles between January 2004 and August 2023 on PPR in small ruminants in Pakistan. Articles were chosen based on specific inclusion and exclusion criteria. A total of 25 articles were selected from 1275 studies gathered from different databases. The overall pooled prevalence in Pakistan was calculated to be 51% (95% CI: 42-60), with heterogeneity I 2 = 100%, τ 2 = 0.0495, and p = 0. The data were summarized based on the division into five regions: Punjab, Baluchistan, KPK, Sindh, and GB and AJK. Among these, the pooled prevalence of PPR in Sindh was 61% (95% CI: 46-75), I 2 = 100%, τ 2 = 0.0485, and p = 0, while in KPK, it was 44% (95% CI: 26-63), I 2 = 99%, τ 2 = 0.0506, and p < 0.01. However, the prevalence of PPR in Baluchistan and Punjab was almost the same. Raising awareness, proper surveillance, and application of appropriate quarantine measures interprovincially and across borders must be maintained to contain the disease.

Background Periodontitis is a chronic inflammatory condition that destroys the tissues supporting the teeth. Conventional nonsurgical treatments, such as mechanical scaling and root surface debridement (RSD), often require adjunct therapies to enhance outcomes due to their limited efficacy in completely eradicating pathogenic microorganisms. Given the adverse effects of standard adjunctive therapies, including antibiotics and nonsteroidal anti-inflammatory drugs, Salvia officinalis (sage) presents a promising herbal alternative due to its anti-inflammatory and antimicrobial properties. This study aims to assess the local application of Salvia officinalis gel as an adjunctive to scaling and RSD to manage periodontitis. Methods We conducted a randomized, controlled split-mouth clinical trial involving 14 systemically healthy periodontitis patients. We included patients with at least 20 natural teeth, a probing pocket depth (PPD) ≥5 mm, and attachment loss ≥4 mm at a minimum of five sites. Primary outcomes measured were bleeding on probing (BOP), PPD, and relative attachment level (RAL). The Salvia officinalis gel was applied to designated test sites post-RSD, while control sites received no adjunctive treatment. Clinical parameters were recorded at baseline and a one-month follow-up visit. Results The cohort consisted of 10 men and four women, with a mean age of 37.1 ± 5.46 years. At the follow-up visit, the test group demonstrated a significant reduction in mean BOP (P = 0.0004), whereas the control group showed no significant change (P ≥ 0.05). Both groups experienced significant decreases in mean PPD and RAL from baseline to follow-up, with the test group showing greater improvements. Conclusions Salvia officinalis gel, used as an adjunct to scaling and RSD, significantly improves clinical periodontal parameters in patients with periodontitis. Its anti-inflammatory properties likely underpin the observed benefits, offering an effective and safe alternative to traditional chemical pharmaceuticals. Further research is needed to explore the long-term effects and mechanisms underlying the therapeutic benefits of Salvia officinalis in periodontal treatment., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2024, Aljuboori et al.)

Manual ground searches and cadaver dogs are traditional methods for locating remains, but they can be time- and resource-intensive, resulting in the decomposition of bodies and delay in victim identification. Therefore, thermal imaging has been proposed as a potentially useful tool for detecting remains based on their temperature. This study investigated the potential of a novel search technique of thermal drones to detect surface remains through the detection of maggot mass temperatures. Two trials were carried out at Selangor, Malaysia, each utilizing 12 healthy male Oryctolagus cuniculus European white rabbits and DJI Matrice 300 RTK drone China, equipped with a thermal camera; Zenmuse H20T to record the thermal imaging footage of the carcasses at various heights (15, 30, 60-100 m) for 14 days for each trial. Our results demonstrated that the larval masses and corresponding heat emissions were at their largest during the active decay stage; therefore, all the carcasses were observable in thermal images on day 5 and remained until day 7. Statistical analyses showed that (1) no statistically significant differences in thermal images between clothed and unclothed subjects (p > 0.05); (2) 15 m above ground level was proven to be the optimal height, as it showed the greatest contrast between the carcass heat signature and the background (p < 0.005). Our data suggested the potential window of detection of thermal signatures was detectable up to 7 days post-deposition. This could be an important guideline for the search and recovery teams for operational implementation in this tropical region., (© 2024 The Authors. Journal of Forensic Sciences published by Wiley Periodicals LLC on behalf of American Academy of Forensic Sciences.)

Zinc oxide nanoparticles (ZnO NPs) have involved a lot of consideration owing to their distinctive features. The ZnO NPs can be described as particularly synthesized mineral salts via nanotechnology, varying in size from 1 to 100 nm, while zinc oxide (ZnO), it is an inorganic substrate of zinc (Zn). The Zn is a critical trace element necessary for various biological and physiological processes in the body. Studies have revealed ZnO NPs' efficient immuno-modulatory, growth-promoting, and antimicrobial properties in poultry birds. They offer increased bioavailability as compared to their traditional sources, producing better results in terms of productivity and welfare and consequently reducing ecological harm in the poultry sector. However, they have also been reported for their toxicological effects, which are size, shape, concentration, and exposure route dependent. The investigations done so far have yielded inconsistent results, therefore, a lot of additional studies and research are required to clarify the harmful consequences of ZnO NPs and to bring them to a logical end. This review explores an overview of efficient possible role of ZnO NPs, while comparing them with other nutritional Zn sources, in the poultry industry, primarily as dietary supplements that effect the growth, health, and performance of the birds. In addition to the anti-bacterial mechanisms of ZnO NPs and their promising role as antifungal, and anti-colloidal agent, this paper also covers the toxicological mechanisms of ZnO NPs and their consequent toxicological hazards to vital organs and the reproductive system of poultry birds., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Background: Timely diagnosis of periodontal disease is crucial for restoring healthy periodontal tissue and improving patients' prognosis. There is a growing interest in using salivary biomarkers as a noninvasive screening tool for periodontal disease. This study aimed to investigate the diagnostic efficacy of two salivary biomarkers, lactate dehydrogenase (LDH) and total protein, for periodontal disease by assessing their sensitivity in relation to clinical periodontal parameters. Furthermore, the study aimed to explore the impact of systemic disease, age, and sex on the accuracy of these biomarkers in the diagnosis of periodontal health., Materials and Methods: A total of 145 participants were categorized into three groups based on their basic periodontal examination index, with 20 in the periodontally healthy group, 50 in the gingivitis group, and 75 in the periodontitis group. Salivary LDH was measured using the rate of nicotinamide adenine dinucleotide (NADH) oxidation, to measure the kinetics of LDH activity, while total protein was measured using the Lowry method. Descriptive and analytical statistical analyses were performed to examine the associations between the variables and biomarkers., Results: The results of the study demonstrated that salivary LDH was 72% sensitive, while salivary total protein was 78% sensitive in correlation to clinical periodontal parameters. The accuracy of the test was not influenced by sex, but age had a significant effect on both biomarkers, particularly LDH. Systemic disease was another factor that significantly affected the accuracy of the test., Conclusions: Although salivary LDH and total protein show promise as biomarkers for screening periodontal disease, their interpretation may be impacted by age and systemic disease. To enhance the accuracy of periodontal disease detection, the study suggests combining these biomarkers with more specific indicators and diagnostic tools., (© 2023 The Authors.)

Carbapenem resistance among Enterobacterales has become a global health concern. Clinical Escherichia coli isolates producing the metallo β-lactamase NDM have been isolated from two hospitals in Faisalabad, Pakistan. These E. coli strains were characterized by MALDI-TOF, PCR, antimicrobial susceptibility testing, XbaI and S1 nuclease pulsed-field gel electrophoresis (PFGE), conjugation assay, DNA hybridization, whole genome sequencing, bioinformatic analysis, and Galleria mellonella experiments. Thirty-four bla NDM producing E. coli strains were identified among 52 nonduplicate carbapenem-resistant strains. More than 90% of the isolates were found to be multidrug resistant by antimicrobial susceptibility testing. S1 PFGE confirmed the presence of bla NDM gene on plasmids ranging from 40 kbps to 250 kbps, and conjugation assays demonstrated transfer frequencies of bla NDM harboring plasmids ranging from 1.59 × 10 -1 to 6.46 × 10 -8 per donor. Whole genome sequencing analysis revealed bla NDM-5 as the prominent NDM subtype with the highest prevalence of bla OXA-1 , bla CTX-M-15 , aadA2 , aac(6')-Ib-cr , and tet(A ) associated resistant determinants. E. coli sequence types: ST405, ST361, and ST167 were prominent, and plasmid Inc types: FII, FIA, FIB, FIC, X3, R, and Y, were observed among all isolates. The genetic environment of bla NDM region on IncF plasmids included partial IS Aba125 , the bleomycin ble gene, and a class I integron. The virulence genes terC, traT, gad, fyuA, irp2, capU, and sitA were frequently observed, and G. mellonella experiments showed that virulence correlated with the number of virulence determinants. A strong infection control management in the hospital is necessary to check the emergence of carbapenem resistance in Gram-negative bacteria.IMPORTANCEWe describe a detailed analysis of highly resistant clinical E. coli isolates from two tertiary care centers in Pakistan including carbapenem resistance as well as common co-resistance mechanisms. South Asia has a huge problem with highly resistant E. coli . However, we find that though these isolates are very difficult to treat they are of low virulence. Thus the Western world has an increasing problem with virulent E. coli that are mostly of low antibiotic resistance, whereas, South Asia has an increasing problem with highly resistant E. coli that are of low virulence potential. These observations allow us to start to devise methodologies to limit both virulence and resistance and combat problems in developing nations as well as the Western world.

Childhood obesity is a significant and growing issue, with the WHO recognising worldwide childhood obesity rates as an epidemic. Primary care is often the first point for monitoring a child's development over time, hence could play an integral part in recognising and addressing childhood obesity. As a result, our systematic review has two objectives. The primary objective is to review the current evidence on best practices in diagnosing and treating childhood obesity. The secondary objective is to review recent qualitative studies looking into the view of primary care practitioners on the treatment and diagnosis of childhood obesity. The rationale for this is to help determine what opportunities there are in primary care in the NHS to tackle childhood obesity. Using searches in MEDLINE, EMBASE, PSYCHINFO, HMIC and NHS evidence over a five-year period from March 2014 to March 2019, a total of 37 studies were eligible for inclusion in the review. Out of these, 25 studies identified exploring the diagnosis and treatment of childhood obesity. A few key themes in these studies were identified, including motivational interviewing, m-health, tools and resources used in consultations, the use of dieticians in the primary care team and factors concerned with the identification of obesity in children. The rest of the 12 qualitative studies involved eliciting the views of direct stakeholders about the diagnosis and treatment of obesity in children. Eight of the studies investigated providers' views towards the role of primary care practitioners in treating childhood obesity, two investigated the parents of obese children's perspectives and the other two investigated general practitioners' (GPs) views towards specific tools and resources. Regarding our primary objective, our findings showed many studies looking at interventions to reduce the BMI in obese children fail to do so in a statistically significant way. However, a few interventions have been more consistent in reducing BMI and obesogenic behaviours. Those interventions include ones utilising the motivational interviewing technique and those targeting families, rather than children. Another key finding was that tools and resources available to primary care providers can significantly impact their ability to diagnose and treat obesity, particularly when looking at the detection. Finally, evidence regarding the clinical effectiveness of e-health is limited, with views on their use also mixed. Regarding our secondary objective, the qualitative research identified demonstrated many common views from GPs across different countries. It showed healthcare providers (HCPs) perceiving the parents as lacking in motivation to address the issue, HCPs not wanting to damage the relationship with their patients due to it being a sensitive topic to bring up, and a lack of time, training and confidence. However, some of these views may not be generalisable to the UK due to cultural and system differences., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2023, Ahmed et al.)

In this research, rabbit femurs were implanted with CP Ti screws coated with a combination of CaCO 3 and nanohydroxyapatite, and the effect on osseointegration was assessed using histological and histomorphometric examination at 2 and 6 weeks. CaCO 3 and nanohydroxyapatite were combined with the EPD to coat the surfaces of the CP Ti screws. The femurs of five male rabbits were implanted with coated and uncoated implant screws. Healing time was divided into two groups (2 and 6 weeks). After 2 and 6 weeks of implantation, the histological examination revealed an increase in the growth of bone cells for coated screws, and the histomorphometric analysis revealed an increase in the percentage of new bone formation (after 6 weeks, 5.08% for coated implants and 3.66% for uncoated implants). In addition, the uncoated implant, the CP Ti implant coated with a combination of CaCO 3 and nanohydroxyapatite, stimulated early bone development after two weeks and mineralization and maturation after six weeks., Competing Interests: The authors declare no conflict of interest., (©2022 JOURNAL of MEDICINE and LIFE.)

Coccidiosis is a protozoan disease that is characterized by diffuse diarrhea, dehydration, emaciation accompanied by moderate morbidity and mild mortality in animals and birds. The current study targeted the molecular characterization of Eimeria isolates in captive deer from different localities in Lahore. The host species was the Cervidae family, such as Hog deer ( Axis porcinus ) and Punjab urial ( Ovis aries vignei ). The Eimeria crandallis was isolated from zoo animals. The DNA was extracted from oocysts and amplified by using reported oligonucleotide primers that exhibited the 809 bp product. These were analyzed by using the small subunit 18S rRNA gene-based evolutionary relationship with 36 other Eimeria species reported in caprine, cervinae, bovines, avians, and rodents. Light microscopic examination exhibited 3.29% (7/213) Eimeria -positive fecal samples with morphological features, including sub-spherical forms, the presence of micropyle with polar cap, and oocysts diameters (μm) ranging from 24.32 ± 1.61 to 18.94 ± 1.51. The phylogenetic tree constitutes four distinct clusters with relatively higher values. The evolutionary network showed that sequences were clustered in the monophyletic group of Eimeria species reported in caprine and cervinae. The nucleotide and amino acid sequence similarity matrix analysis exhibited 99.5-99.9% identity of the study isolates with Eimeria crandallis (AF336339). This study provides relevant baseline data to develop strategic control measures for coccidiosis in zoo animals. However, further investigations are required to place the hog deer and Punjab urial-derived E. crandallis into the caprine-originated cluster., Competing Interests: The authors declare no conflicts of interest.

LHPP gene encodes a phospholysine phosphohistidine inorganic pyrophosphate phosphatase, which functions as a tumor-suppressor protein. The tumor suppression by this protein has been confirmed in various cancers, including hepatocellular carcinoma (HCC). LHPP downregulation promotes cell growth and proliferation by modulating the PI3K/AKT signaling pathway. This study identifies potentially deleterious missense single nucleotide variants (SNVs) associated with the LHPP gene using multiple computational tools based on different algorithms. A total of 4 destabilizing mutants are identified as L22P, I212T, G227R , and G236R , from the conserved region of the phosphatase. The 3-dimensional (3D) modeling and structural comparison of variants with the native protein reveals significant structural and conformational variations after mutations, suggesting disruption in the function of phospholysine phosphohistidine inorganic pyrophosphate phosphatase. The identified mutations might, therefore, participate in the cause of HCC., Competing Interests: Declaration Of Conflicting Interests: The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (© The Author(s) 2022.)

Protein engineering and enzyme immobilization strategies have produced numerous biocatalysts for modern industrial applications. In this study, we have also used these two strategies for improving the operational stability and catalytic efficiency of serine protease from Pseudomonas aeruginosa. The enzyme serine protease was truncated to separate its trypsin-like domain from the PDZ1 and PDZ2 domains. The truncated trypsin-like domain was expressed in Escherichia coli BL21, and its catalytic activity and thermostability were estimated. Later this trypsin-like domain was immobilized with 2% Na-alginate. The immobilized domain showed 10°C increase in optimum temperature compared to its free counterpart. Kinetic studies showed two-folds increased V max of the immobilized domain. Likewise, the K m value of this domain was 11.5 folds lower compared to the free trypsin-like domain. The catalytic efficiency (K cat /K m ) of the immobilized enzyme also elevated to 311 folds. Additionally, the immobilized trypsin-like domain remained active in the presence of surfactants (Triton-X 100, SDS, and Tween-40) and metal ions (Mg 2+ , Ca 2+ , Na + , and Zn 2+ ). It also efficiently removes gelatin layer from X-ray film and hair from sheepskin. Thus, the immobilized trypsin-like domain of serine protease, with increased thermostability and catalytic efficiency, is operationally more stable than the soluble truncated trypsin-like domain., (© 2022 Wiley Periodicals LLC.)

The coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is threatening public health. A large number of affected people need to be hospitalized. Immunocompromised patients and ICU-admitted patients are predisposed to further bacterial and fungal infections, making patient outcomes more critical. Among them, COVID-19-associated candidiasis is becoming more widely recognized as a part of severe COVID-19 sequelae. While the molecular pathophysiology is not fully understood, some factors, including a compromised immune system, iron and zinc deficiencies, and nosocomial and iatrogenic transmissions, predispose COVID-19 patients to candidiasis. In this review, we discuss the existing knowledge of the virulence characteristics of Candida spp. and summarize the key concepts in the possible molecular pathogenesis. We analyze the predisposing factors that make COVID-19 patients more susceptible to candidiasis and the preventive measures which will provide valuable insights to guide the effective prevention of candidiasis in COVID-19 patients., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Buffalopox (BPX) is a highly contagious disease that causes high morbidity and production losses in buffaloes. During this study, seroprevalence, effect of various associated risk factors, and pathological studies of BPX were recorded in the Punjab province. A total of 97 blood samples and 63 scabs were collected from clinically pox suspected buffaloes. Serum was harvested to perform single radial hemolysis to assess the seroprevalence, and scabs were subjected to PCR for BPX virus confirmation. Results revealed that, animal demographics and environmental associated factors showed significant effect (p⟨0.05,1⟨R2⟩0) on BPX occurrence. The overall BPX seroprevalence was recorded 4.18% in the Punjab province. The BPX was recorded 5.48% in Nili Ravi breed during winter (7.42%), aged 5-7 years (7.46%) under loose housing (5.51%) in the Faisalabad region (8.03%). Further, BPX was 5.37% in pregnant, 6.86% pregnant milking buffaloes during the 3rd lactation period (7.28%) in dairy herds (5.20%). The BPX was 5.22% in non-vaccinated buffaloes where multiple animals were reared together (4.99%) in the herds having 21-30 total number of animals. A total of 49 scab samples were found positive for the BPX virus via PCR with C18L gene amplification. Grossly, inflammatory lesions with pits in the center and wart-like nodules were seen on teats and udder of buffaloes. Increased leukocytes, especially neutrophils and lymphocytes, were seen in the blood of the infected animals. These results provide a broader window to understand the effect of associated risk factors, strengthen the diagnostic aid, and to contain the current spread of BPX in Pakistan to safeguard large ruminant-based livelihood., (Copyright© by the Polish Academy of Sciences.)

Alzheimer's disease (AD) is the most common neurodegenerative disorder worldwide. Ongoing research to develop AD treatments has characterized multiple drug targets including the cholinergic system, amyloid-β peptide, phosphorylated tau, and neuroinflammation. These systems have the potential to interact to either drive or slow AD progression. Promising agents that simultaneously impact many of these drug targets are the AD experimental drug Posiphen and its enantiomer phenserine that, currently, are separately being evaluated in clinical trials. To define the cholinergic component of these agents, the anticholinesterase activities of a ligand dataset comprising Posiphen and primary metabolites ((+)-N1-norPosiphen, (+)-N8-norPosiphen, and (+)-N1,N8-bisnorPosiphen) were characterized and compared to those of the enantiomer phenserine. The "target" dataset involved the human cholinesterase enzymes acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). Binding interactions between the ligands and targets were analyzed using Autodock 4.2. The computationally determined inhibitory action of these ligands was then compared to ex vivo laboratory-measured values versus human AChE and BChE. While Posiphen lacked AChE inhibitory action, its major and minor metabolites (+)-N1-norPosiphen and (+)-N1,N8-bisnorPosiphen, respectively, possessed modest AChE inhibitory activity, and Posiphen and all metabolites lacked BChE action. Phenserine, as a positive control, demonstrated AChE-selective inhibitory action. In light of AChE inhibitory action deriving from a major and minor Posiphen metabolite, current Posiphen clinical trials in AD and related disorders should additionally evaluate AChE inhibition; particularly if Posiphen should be combined with a known anticholinesterase, since this drug class is clinically approved and the standard of care for AD subjects, and excessive AChE inhibition may impact drug tolerability., Competing Interests: The authors declare no competing financial interest., (© 2022 The Authors. Published by American Chemical Society.)

The use of drones has rapidly evolved over the past decade involving a variety of fields ranging from agriculture, commercial and becoming increasingly used in disaster management or humanitarian aid. Unfortunately, the evidence of its use in mass disasters is still unclear and scarce. This article aims to evaluate the current drone feasibility projects and to discuss a number of challenges related to the deployment of drones in mass disasters in the hopes of empowering and inspiring possible future work. This research follows Arksey and O'Malley framework and updated by Joanna Briggs Institute Framework for Scoping Reviews methodology to summarise the results of 52 research papers over the past ten years, from 2009 to 2020, outlining the research trend of drone application in disaster. A literature search was performed in Medline, CINAHL, Scopus, individual journals, grey literature and google search with assessment based on their content and significance. Potential application of drones in disaster are broad. Based on articles identified, drone application in disasters are classified into four categories; (1) mapping or disaster management which has shown the highest contribution, (2) search and rescue, (3) transportation and (4) training. Although there is a significant increase in the number of publications on use of drone in disaster within the last five years, there is however limited discussion to address post-disaster healthcare situation especially with regards to disaster victim identification. It is evident that drone applications need to be further explored; to focus more on drone assistance to humans especially in victim identification. It is envisaged that with sufficient development, the application of drones appears to be promising and will improve their effectiveness especially in disaster management., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: The British Thoracic Society (BTS) recommends that all patients admitted with COVID-19 pneumonia should have a chest X-ray (CXR) and clinical follow-up at 6 or 12 weeks, depending on the disease severity. Little data is available on long-term CXR follow-up for moderate and severe COVID-19 pneumonia. This study aims to evaluate compliance with clinico-radiological follow-up of patients recovering from COVID-19 pneumonia at a local hospital in the UK, as per the BTS guidance, and to analyse radiological changes at clinical follow-up at 12 weeks, in order to risk-stratify and improve patient outcomes., Methods: This is a single-centre retrospective audit of 255 consecutive COVID-19 positive patients admitted to a local hospital in the UK over 5 months between May and October 2020. All CXRs and clinic follow-up at 12 ± 8 weeks were checked on an electronic database., Results: Over one in two (131/255) patients had CXR evidence of COVID-19 pneumonia during the initial hospital admission. Half of the patients (60/131) died before CXR or clinic follow-up. Fifty-eight percent (41/71) of the surviving patients had a follow-up CXR, and only two developed respiratory complications- one had residual lung fibrosis, another a pulmonary embolism. Eighty-eight percent (36/41) of the patients had either resolution or improved radiological changes at follow-up. Most patients who had abnormal follow-up CXR were symptomatic (6/8), and many asymptomatic patients at follow-up had a normal CXR (10/12)., Conclusions: Although there were concerns about interstitial lung disease (ILD) incidence in patients with COVID-19 pneumonia, most of our patients with COVID-19 pneumonia had no pulmonary complications at follow-up with CXR. This emphasises that CXR, a cost-effective investigation, can be used to risk-stratify patients for long term pulmonary complications following their COVID-19 pneumonia. However, we acknowledge the limitations of a low CXR and clinic follow-up rate in our cohort., (© 2021. The Author(s).)

It has been a difficult year ahead since the announcement of COVID-19 as a pandemic by the Director General of the World Health Organization (WHO). Many countries including Malaysia have struggled to take urgent and aggressive measures to advise, detect, screen, test, isolate and treat their citizens in order to fight against the virus. All countries must strike a balance between protecting health, minimizing economic and social disruption and at the same time, preserving human rights which includes managing the dead with dignity and respect. The National Institute of Forensic Medicine (IPFN) Malaysia has been proactively engaging the various agencies involved in a concerted effort to ensure smooth flow in the management of the dead for all COVID-19 and non-COVID-19 cases since the beginning of the outbreak in February 2020. This is in accordance with all existing guidelines and regulations vis-a-vis infection prevention and control in the country. However, new situations that arise during the pandemic are always challenging and dynamic. The COVID-19 crisis has uncovered a plethora of issues surrounding the margin of the Forensic Medicine fraternity. Therefore, this article highlights various emerging issues encountered by the IPFN and how they are resolved through good leadership to guide and direct operations and activities during the exponential rise of COVID-19 infection in Malaysia. In the provision of the management of the dead, it is of utmost importance to note that the humanitarian aspect needs to be emphasized even in times of an outbreak to give closure to the family and the opportunity to mourn their loved ones., (© 2021 The Authors.)

Objectives: The design of peptide-based vaccines for cancer is a promising immunotherapy that can induce a cancer-specific cytotoxic response in tumor cells., Methods: Herein, we used the immunoinformatic approach in designing a multi-epitope vaccine targeting G-protein coupled receptor 87 (GPCR-87), cystine/glutamate transporter (SLC7A11), Immunoglobulin binding protein 1 (IGBP1), and thioredoxin domain-containing protein 5 (TXNDC5), which can potentially contribute to NSCLC. The MHC-I and MHC-II epitopes selected for the fusion construct were evaluated for their antigenic and non-allergenic natures via VaxiJen and AllerTop., Results: A total of five epitopes, four class-I (FIFYLKNIV, CRYTSVLFY, RYLKVVKPF, and RQAKIQRYK), and one class-II (NQVRGYPTLLWFRDG), having combined USA population coverage of 100%, were used to make ten possible multi-epitope fusion constructs. In these constructs, PADRE, a universal T-helper epitope, and RSO9, a TLR4 agonist, were fused as adjuvants. The molecular docking analysis revealed that two constructs were showing significant binding affinities toward HLA -A*02:01, the most prevalent HLA allele in USA. Moreover, MD simulations marked one construct as a promising therapeutic candidate., Conclusion: The multi-epitope vaccine constructs designed using immunogenic, and non-allergenic peptides of NSCLS tumor-associated proteins are likely to pose significant therapeutic efficacies in cancer immunotherapy due to their high binding affinities toward HLA molecules.

Background: Blast related deaths are often shrouded by diagnostic and medicolegal complexities requiring multidisciplinary expertise in order to gauge accurate identification of the victims and document scientific investigations comprehensively. In the advent of more sophisticated technology, anthropologic methods can now be applied into post mortem imaging interpretation. The traditional imaging roles of characterizing osseous fragmentation, detecting and localizing foreign bodies can be expanded to simulate and support physical anthropologic examination to assist in documentation for court proceedings., Case Presentation: An assemblage of unidentified, incomplete, highly fragmented skeletal remains were found scattered on a bare area of land in a forest. There was evidence of an explosion given the pattern of scattered evidentiary material of explosive and ballistic nature. Laboratory analysis of white powder found within the explosive material confirmed the presence of high impact C4-explosive trace containing cyclotrimethylene trinitramin [Royal Demolition Explosive (RDX)] & pentaerythritol tetranitrate (PETN). It took meticulous multidisciplinary efforts to confirm the identity of the victim that was marred by the severe fragmentation and skeletalization of the remains. The initial radiologic interpretation focused more on identification of foreign bodies and supporting documentation of fragmentation. With the current availability of post computed tomography (PMCT) in our center, we reexamined the value and potential of PMXR and PMCT as an adjunctive tool for biological profiling., Conclusion: This was the first case of C4-blast related death reported in Malaysia. The multidisciplinary approach in efforts to identify the victim may serve as a guide in managing, coordinating and maximizing the expertise of different forensic specialists, with emphasis on anthropologic and radiologic collaboration., (Copyright © 2021. Published by Elsevier Ltd.)

Increasing trend in antimicrobial resistance and failure of chemically synthesized antibiotics lead to discover alternative methods for the treatment of bacterial infections. Various medicinal plants are in use traditionally and their active compounds can be further applied for treatment of bacterial diseases. This study was designed to determine the antibacterial activity of Punica granatum (P. granatum L.) (pomegranate) peel extract against Enterobacteriaceae [Escherichia coli (E. coli), Salmonella Typhimurium (S. Typhimurium) and Shigella Dysenteriae (S. Dysenteriae)] and gram-positive bacterium [Staphylococcus aureus (Staph aureus)]. Methanolic extract of P. granatum L. peel was prepared by Soxhlet apparatus method. Total flavonoid and phenolic contents from the extract were determined by High Performance Liquid Chromatography (HPLC). The antibacterial activity of P. granatum L. peel extract was evaluated through agar well diffusion method. HPLC showed the range of phenolics (gallic acid, caffeic acid, benzoic acid, cinnamic acid) and flavonoid compounds. The chemical structures of flavonoid and phenolics found in the methanolic extract of P. granatum L. peel have been reported for the first time. The methanolic peel extract (50 ul) of yellow P. granatum L. showed 26, 10, 10 and 9mm zones of inhibition (ZOI) against S. aureus, S. Typhimurium, S. Dysenteriae and E. coli, respectively. The methanolic extract of red P. granatum L. (100 ul) showed 27, 8, 12 and 15 mm ZOI against Staph. aureus, S. Typhimurium, S. Dysenteriae and E. coli, respectively. Highest ZOI was observed against Staph. aureus. Many of the bacteria studied in the present work may cause serious gastrointestinal infections, which can lead to hemorrhagic diarrhea in children. These infections can be life-threatening to young children and the elderly. There is an incentive to find alternative control measures, such as plant and herbal extracts, especially in lesser-developed countries where traditional antibiotics may not be readily available.

Since the end of 2019, the emergence of novel coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has accelerated the research on host immune responses toward the coronaviruses. When there is no approved drug or vaccine to use against these culprits, host immunity is the major strategy to fight such infections. Type I interferons are an integral part of the host innate immune system and define one of the first lines of innate immune defense against viral infections. The in vitro antiviral role of type I IFNs against Middle East respiratory syndrome coronavirus (MERS-CoV) and SARS-CoV (severe acute respiratory syndrome coronavirus) is well established. Moreover, the involvement of type I IFNs in disease pathology has also been reported. In this study, we have reviewed the protective and the immunopathogenic role of type I IFNs in the pathogenesis of MERS-CoV, SARS-CoV, and SARS-CoV-2. This review will also enlighten the potential implications of type I IFNs for the treatment of COVID-19 when used in combination with IFN- γ .

Currently, the whole world is facing the coronavirus disease-19 pandemic. As of now, approximately 0.15 million people around the globe are infected with the novel coronavirus. In the last decade, two strains of the coronavirus family, severe acute respiratory syndrome-related coronavirus and Middle East respiratory syndrome coronavirus, also resulted in epidemics in south Asian and the Middle Eastern countries with high mortality rate. This scenario demands the development of a putative vaccine which may provide immunity against all current and new evolving coronavirus strains. In this study, we designed an epitope-based vaccine using an immunoinformatic approach. This vaccine may protect against all coronavirus strains. The vaccine is developed by considering the geographical distribution of coronavirus strains and host genetics (Chinese population). Nine experimentally validated epitopes sequences from coronavirus strains were used to derive the variants considering the conservancy in all strains. Further, the binding affinities of all derived variants were checked with most abundant human leukocyte antigen alleles in the Chinese population. Three major histocompatibility complex (MHC) Class I epitopes from spike glycoprotein and nucleoprotein showed sufficient binding while one MHC Class II epitope from spike glycoprotein was found to be an effective binder. A cocktail of these epitopes gave more than 95% population coverage in the Chinese population. Moreover, molecular dynamics simulation supported the aforementioned predictions. Further, in vivo studies are needed to confirm the immunogenic potential of these vaccines., (© 2021 The Societies and John Wiley & Sons Australia, Ltd.)

Objective: To determine the association of Upper extremity musculoskeletal disorders and Exposure to Ergonomic risk factors among handicraft workers., Methods: This cross-sectional survey was conducted in a 12 weeks' duration i.e. from November 2018 till January 2019. Nordic musculoskeletal questionnaire was used to determine the frequency of upper limb musculoskeletal disorders. Postural analysis was done using Rapid Upper Limb Assessment (RULA). Data was collected from 100 Handicraft workers and the type of work included Art work (n=18), Ada Work (n=7), botanical arrangement (n=11), Textile, fashion designing and stitching (n=38), Fine arts (n=20), embroidery and knitting (n=6)., Results: The frequency distribution of Rapid upper limb assessment (RULA) score for exposure to risk factors showed that 47 participants were at a high risk and required implementation of change. Most of the participants (n=35) reported pain in neck and shoulder (n=29). The chi square test for association between pain in upper extremity and exposure to risk factors showed that only wrist pain had statistically significant association with overall RULA score (p-value<0.05)., Conclusion: The study concluded that neck and shoulder are more frequently affected among handicraft workers. The frequency of workers who were at high ergonomic risk and required implementation of change in working conditions was high., Competing Interests: Conflicts of Interest: None., (Copyright: © Pakistan Journal of Medical Sciences.)

Coronavirus Disease 2019 (COVID-19) ranks third in terms of fatal coronavirus diseases threatening public health, coming after SARS-CoV (severe acute respiratory syndrome coronavirus), and MERS-CoV (Middle East respiratory syndrome coronavirus). SARS-CoV-2 (severe acute respiratory syndrome coronavirus type 2) causes COVID-19. On January 30, 2020, the World Health Organization (WHO) announced that the current outbreak of COVID-19 is the sixth global health emergency. As of December 3, 2020, 64 million people worldwide have been affected by this malaise, and the global economy has experienced a loss of more than $1 trillion. SARS-CoV-2 is a positive-sense single-stranded RNA virus belonging to the Betacoronavirus genus. The high nucleotide sequence identity of SARS-CoV-2 with the BatCoV RaTG13 genome has indicated that bats could be the possible host of SARS-CoV-2. SARS-CoV-2 penetrates the host cell via binding its spike protein to the angiotensin-converting enzyme 2 (ACE2) receptor, which is similar to the mechanisms of SARS-CoV and MERS-CoV. COVID-19 can spread from person to person via respiratory droplets and airborne and contaminated fomites. Moreover, it poses a significant risk to smokers, the elderly, immunocompromised people, and those with preexisting comorbidities. Two main approaches are used to control viral infections, namely, vaccination, and biosecurity. Studies to analyze the antigenicity and immunogenicity of SARS-CoV-2 vaccine candidates are underway, and few vaccines may be available in the near future. In the current situation, the Human Biosecurity Emergency (HBE) may be the only way to cope effectively with the novel SARS-CoV-2 strain. Here, we summarize current knowledge on the origin of COVID-19 as well as its epidemiological relationship with humans and animals, genomic resemblance, immunopathogenesis, clinical-laboratory signs, diagnosis, control and prevention, and treatment. Moreover, we discuss the interventional effects of various nutrients on COVID-19 in detail. However, multiple possibilities are explored to fight COVID-19, and the greatest efforts targeted toward finding an effective vaccine in the near future. Furthermore, antioxidants, polyphenols, and flavonoids, both synthetic and natural, could play a crucial role in the fight against COVID-19., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Siddique, Abbas, Mansoor, Alghamdi, Saeed, Ayaz, Rahman, Mahmood, Iqbal, Manzoor, Abbas, Javaid and Hussain.)

Tuberculosis (TB), a chronic disease caused by  Mycobacterium tuberculosis (Mtb) , is a global health issue across the world. Pakistan ranks fifth among the countries, which are facing, a significantly great number of mortalities and morbidities due to TB. Unfortunately, all previously reported treatments are not successful for the eradication of TB. Here in this study, we report an emerging treatment option for this disease. We have applied immunoinformatics to predict highly conserved B and T-cell epitopes from Mtb , showing significant binding affinities to the frequent HLA alleles in the Pakistani population. A total of ten highly referenced and experimentally validated epitopes were selected from the Immune Epitope Database (IEDB), followed by their conservancy analysis using weblogos. The consensus sequences and variants derived from these sequences were examined, for their binding affinities, with prevalent HLA alleles of Pakistan. Moreover, the antigenic and allergenic natures of these peptides were also evaluated via Vaxijen and AllerTOP, respectively. Consequently, all potentially allergenic and non-antigenic, peptide fragments, were excluded from the analysis. Among all putative epitopes, three CD8 + T-cell epitopes were selected, as ideal vaccine candidates and, population coverage analysis revealed that the combination of these three peptides was covering, 67.28% Pakistani Asian and 57.15% mixed Pakistani populations. Likewise, eleven linear and six conformational or discontinuous B-cell epitopes were also marked as potential vaccine candidates based on their prediction score, non-allergenic nature, and antigenic properties. These epitopes, however, need the final validation via wet-lab studies. After their approval, these epitopes would be effective candidates for the future designing of epitope-based vaccines against Mtb infections in Pakistan., Competing Interests: Conflict of interestAll authors declare that they have no conflict of interest., (© Springer Nature B.V. 2020.)

The emergence of a novel human coronavirus, SARS-CoV-2, causing severe respiratory tract infections in humans, is affecting all countries of the world and has become a global health concern. Since the virus was first identified in December 2019, the number of deaths have been propagating exponentially, causing countries across the world, including Malaysia, to increase emergency measures to combat the virus. Due to the fact that the COVID-19 pandemic does not discriminate its victims, it is of paramount importance to construct a plan for management of the dead for all suspected or confirmed COVID-19 cases, including the unidentified deceased, as an essential portion of the humanitarian forensic action approach. This document provides an overview on ways to maximize the local collective capacity from various government agencies to manage the dead based on the prevailing regulations and legislation in the country, in preparation for possible large scale deaths from this pandemic. The National Institute of Forensic Medicine Malaysia has improvised procedures and guidelines for management of the dead within the existing regulations in order to achieve a balance between medicolegal requirements and the safety of personnel managing the bodies of the deceased with suspected or confirmed COVID-19 infection; at the site of death, during transport, during postmortem procedures, storage and preparation before and during burial or cremation as well as environmental cleaning and disinfection, involving various agencies in the country. A form of temporary controlled burial is as an option to allow the reinvestigation of a decedent to help formally identify victims of the pandemic such as undocumented migrants or refugees who were previously not identified. Due to the different legal requirements and mortality rates between countries, there is no one-size-fits-all approach to the management of the dead. Whenever possible, every opportunity and assistance must be given to families to mourn their loved ones, even in times of crisis or an outbreak, in order to sustain an appropriate level of dignity and respect.

During a disease pandemic, there is still a requirement to perform postmortem examinations within the context of legal considerations. The management of the dead from COVID-19 should not impede the medicolegal investigation of the death where required by the authorities and legislation but additional health and safety precautions should be adopted for the necessary postmortem procedures. The authors have therefore used the craniotomy box in an innovative way to enable a safe alternative for skull and brain removal procedures on suspected or confirmed COVID-19 bodies. The craniotomy box technique was tested on a confirmed COVID-19 positive body where a full postmortem examination was performed by a team of highly trained personnel in a negative pressure Biosafety Level 3 (BSL-3) autopsy suite in the National Institute of Forensic Medicine (IPFN) Malaysia. This craniotomy box is a custom-made transparent plastic box with five walls but without a floor. Two circular holes were made in one wall for the placement of arms in order to perform the skull opening procedure. A swab to detect the presence of the SARS-CoV-2 virus was taken from the interior surface of the craniotomy box after the procedure. The result from the test using real-time reverse transcriptase polymerase chain reaction (rRT-PCR) proved that an additional barrier provided respiratory protection by containing the aerosols generated from the skull opening procedure. This innovation ensures procedures performed inside this craniotomy box are safe for postmortem personnel performing high risk autopsies during pandemics.

Competing Interests: The authors declared that they have no conflict of interest.

Background: Neonatal sepsis is a leading cause of morbidity and early-life mortality worldwide, and previous data have reported the highest neonatal mortality rate in Pakistan., Purpose: The present study aimed to decipher the prevalence of group B Streptococcus (GBS)-associated sepsis, coinfections, and antibiotic susceptibility of isolated microbes in neonates., Methods: Blood samples of 100 cases of neonatal sepsis were subcultured on blood agar, GBS agar, and MacConkey agar for isolation of GBS and suspected microbes., Results: Of 100 neonatal blood samples, 85 blood samples were culture-positive, including mixed culture growth (n = 18), 25 samples as early-onset neonatal sepsis (EONS) and 60 as late-onset neonatal sepsis (LONS). Staphylococcus aureus showed high percent positivity (31.4%), followed by Candida sp (16.5%), GBS (14.1%), Klebsiella (14.1%), Staphylococcus epidermidis (11.8%), Pseudomonas (9.4%), Acinetobacter (9.4%), Esherichia coli (8.2%), and Enterococcus (5.9%). GBS was isolated more frequently from EONS than from LONS with 50% coinfections. Mode of delivery, gender, and respiratory distress in neonates were significantly associated with culture-positive sepsis. GBS isolates were highly (91.7%) susceptible to vancomycin, cefotaxime, and chloramphenicol, followed by penicillin (83.3%), ampicillin, and tetracycline (75%). GBS isolates were resistant to erythromycin, clindamycin, ciprofloxacin, and linezolid., Implications for Practice: Our findings evidenced GBS-associated risk factors and antibiotic susceptibility pattern of neonatal sepsis, which will help clinicians in management, control, and treatment of neonatal sepsis., Implications for Research: The epidemiological evidence of GBS-associated neonatal sepsis, demographic characteristics, risk factor data analysis, and drug resistance pattern has disease prevention implications in neonates in low-income countries including Pakistan.

Objective: The aim of study was to investigate the association of IL 1B gene polymorphism with involvement of H. pylori and other gastric diseases., Methods: Blood samples of dyspeptic patients were collected from endoscopy department of Allied Hospital Faisalabad from January 2017 to January 2019 and were qualitatively assayed for serological detection of CagA H. pylori antibodies. PCR followed by direct sequencing was performed for proinflammatory IL-1B gene polymorphism detection. Sequence analysis was performed in software SnapGene viewer for haplotypes., Results: Demographic characteristics of seropositive patients showed maximum 25% gastritis in age groups of 20-40 years and 41-60 years, predominantly (41.7%) in females. While in seronegative patient's gastritis (33.3%) was found in age group of 20-40 years mainly in males (41.7%). Among studied groups, higher expression of IL-1B-511 genotype (33.3%) polymorphism was found in healthy individuals as compared to H. pylori seropositive (25%) and seronegative (8.3%). While IL-1B-31 genotype showed maximum 33.3% polymorphism rate in seropositive gastric diseased group. Moreover, haplotypes frequencies IL-1B-511CC and IL-1B-31TT were predominantly (20%) found in seropositive gastric diseased group., Conclusions: In H. pylori seropositive patients, gastric disease was commonly found, however, gastric disease was not only associated with H. pylori as seronegative patients were also carrying gastric complications. Interleukin IL-1B polymorphism was partially associated with H. pylori infection in studied dyspeptic population., Competing Interests: Conflicts of interest: None., (Copyright: © Pakistan Journal of Medical Sciences.)

Introduction: Methanol is a widely available chemical with a range of uses including as solvent, as a fuel, in chemical synthesis and anti-freeze preparations. Most of the cases are accidental exposures to drinking beverages contaminated with methanol., Materials and Methods: In mid-September 2018, there was a single outbreak of methanol poisoning in Malaysia especially involving the state of Federal Territory Kuala Lumpur and Selangor. There were 33 reported deaths suspected due to methanol poisoning in this current outbreak where 11 of them were brought in to the Institute of Forensic Medicine (NIFM), Kuala Lumpur. The last outbreak was in the year 2013 with 29 deaths reported out of 44 cases., Results: There were 3 cases (27.2%) died in hospital and the remaining 8 cases (72.8%) were found dead at home and were later brought in dead to the hospital. A full autopsy was carried out for each case. Autopsy findings, as well as lab results pertaining to cases that survived and directly brought in dead, were of a different spectrum., Conclusion: Methanol related deaths are almost always as a result of greed. The running truism is 'methanol poisoning is a result of deliberate addition/adulteration with industrial methanol'. Prevention of the illegal production of methanol and methylated spirits should be established to curb this matter in the future.

Background: Post-mortem Computed Tomography (PMCT) allows non-invasive or minimally invasive detection of findings that may or may not be visible during conventional autopsy, however, it does not allow the investigator to draw any conclusions regarding patency of the vessel's lumen. To address this deficiency, Post-mortem Computed Tomography Angiography (PMCTA) utilizing different contrast media and techniques have been introduced with various studies looking at the correlation between PMCTA, autopsy (gross) findings and coronary artery histology in diagnosing coronary artery disease., Objectives: The aim of this study is to investigate the sensitivity and specificity of PMCTA in diagnosing coronary artery stenosis using water-based contrast media introduced though the vessels of the neck, compared to the gold standard of diagnosis i.e. gross and histological evaluation of the coronary artery., Method: This was a cross sectional study of 158 arterial sections involving 37 subjects recruited from the National Institute of Forensic Medicine (IPFN), Hospital Kuala Lumpur (HKL). An unenhanced PMCT was performed followed by PMCTA using water-based contrast media introduced though the vessels of the neck. Coronary artery stenosis was determined using multiplanar reconstructionD while the degree of stenosis was determined by calculating the percentage of luminal diameter divided by the diameter of the vessel internal elastic., Results: The analysis of PMCTA and histopathology examinations revealed a sensitivity of 61.5%, specificity of 91.7%; positive predictive value (PPV) of 40.0% and negative predictive value (NPV) of 96.4%., Conclusion: PMCTA utilizing water-based contrast introduced though the vessels of the neck yielded similar results as other methods and techniques of PMCTA. We would therefore conclude that PMCTA utilizing this technique could be used to assess the degree of calcification and the presence of significant stenosis., Competing Interests: Declaration of Competing Interest The authors of this manuscript do not believe that there is a conflict of interest that could potentially be construed to affect the material contained in the manuscript that is being submitted to the Journal., (Copyright © 2020 Elsevier B.V. All rights reserved.)

This study explores the practicability and usability of the cling film plastic wrap on the dead body with decomposition changes, whereby the feasibility of wrapping the remains to ensure the containment of the decomposition fluid within the cling film plastic wrap. Unknown and unclaimed dead bodies were used. Wrapping and preservation of human remains using the cling film plastic wrap could be the best operational practices for first responders rather than leaving bodies exposed on the disaster site, when the supplies of the cadaver body bags are inadequate. Thinking out of the box, the conventional way of using cadaver body bags to the cling film plastic wrap forms a new perspective in managing the dead and facilitating the human identification needs. New inventive idea of adopting the cling film plastic wrap as means of protecting the dignity of the dead person, could be the way forward in humanitarian forensic action., Competing Interests: The authors declare that they have no competing interests., (© 2020 The Authors. Published by Elsevier B.V.)

Pectinase enzymes from different plants have many possible biotechnological applications in various industries. Considering industrial significance of pectinolytic enzymes, the polygalacturonase (PG) gene from grapes was cloned into Escherichia coli DH5α using pTZ57R/T vector. Homologous sequences established a close link to Vitis vinifera. Conserve domain analysis predicted PLN02218 domain belongs to the cl31843 superfamily, showing its function as polygalacturonase. After confirmation by PCR and restriction analysis, the PG gene was expressed in E. coli BL21 and induced by IPTG. Expression level was assessed by 12% SDS-PAGE, which showed a 47 kDa protein. High expression level in the soluble fraction indicated that the protein is intracellular or transmembrane. Maximum expression was obtained with 1 mM IPTG and 6 hour induction time, and autoinduction with lactose increased production of the recombinant enzyme. Zymogram analysis revealed that the induced protein was an active enzyme. Expressed PG showed pectinolytic effect on the physiochemical properties of lemon juice. Natural biopolymers are greatly needed because synthetic fibers can negatively affect health. Pectin hydrolysis of banana pseudostem by the PG enzyme produced better quality fiber. Morphological studies by scanning electron microscopy revealed pectin degradation within the fiber cell architecture, showing the effectiveness of PG treatment on banana pseudostem.

The use of interferon α-2 in combination with thymosin α-1 shows higher anti-cancer effect in comparison when both are used individually because of their synergistic effects. In this study we produced an important human interferon α-2-thymosin α-1 (IFNα2-Tα1) fusion protein with probable pharmaceutical properties coupled to its high-level expression, characterization, and study of its biological activity. The IFNα2-Tα1 fusion gene was constructed by over-lap extension PCR and expressed in Escherichia coli expression system. The expression of IFNα2-Tα1 fusion protein was optimized to higher level and its maximum expression was obtained in modified terrific broth medium when lactose was used as inducer. The fusion protein was refolded into its native biologically active form with maximum yield of 83.14% followed by purification with ∼98% purity and 69% final yield. A band of purified IFNα2-Tα1 fusion protein equal to ∼23 kDa was observed on 12 % SDS-PAGE gel. The integrity of IFNα2-Tα1 fusion protein was confirmed by western blot analysis and secondary structure was assessed by CD spectroscopy. When IFNα2-Tα1 fusion protein was subjected to its biological activity analysis it was observed that it exhibits both IFNα2 & Tα1 activities as well as significantly higher anticancer activity as compared to IFNα-2 alone.

Infective endocarditis (IE) is a relatively uncommon disease, but has been challenging to diagnose over the years. With the increasing incidence, variety of causative agents and the resistance of microorganisms towards antibiotics, there is still an occurrence of sudden death due to undiagnosed IE. The most common microorganism causing IE is Staphylococcus aureus. However, there is increasing prevalence of other microorganisms causing IE. This case report highlights a case of sudden death due to IE caused by a rare pathogen, Streptococcus constellatus which belongs to the Streptococcus anginosus group (Milleri group). A study noted the crude incidence of IE in 6 world regions ranged between 1.5 and 11.6 cases per 100,000 people. To date, there has been no previous report on sudden death due to IE caused by Streptococcus constellatus in Malaysia, neither in the forensic nor clinical setting. This case report underlined the characteristics and pathological features of this microorganism. The increasing incidence and variety of causative organisms in IE are important public health issues. It is vital for future studies to examine the risk factors of IE related to Streptococcus constellatus, to enhance better understanding, insight and awareness regarding the course of this disease. This in turn may facilitate preventive measures to avoid morbidity and mortality from this condition.

Increased demand of enzymes for industrial use has led the scientists towards protein engineering techniques. In different protein engineering strategies, rational approach has emerged as the most efficient method utilizing bioinformatics tools to produce enzymes with desired reaction kinetics; physiochemical (temperature, pH, half life, etc) and biological (selectivity, specificity, etc.) characteristics. Xylanase is one of the widely used enzymes in paper and food industry to degrade xylan component present in plant pulp. In this study endo 1,4-β-xylanase (Xyl-11A) from Bacillus halodurans C-125 was cloned in pET-22b (+) vector and expressed in Escherichia coli BL21 (DE3) expression strain. The enzyme had Michaelis constant K m of 1.32 mg ml -1 birchwoodxylan (soluble form) and maximum reaction velocity (V max ) 73.53 mmol min -1  mg -1 with an optimum temperature of 75 °C and pH 9.0. The thermostability analysis showed that enzyme retained more than 80% of its residual activity when incubated at 75 °C for 2 h. In addition, to increase Xyl-11A thermostability, an in-silico analysis was performedto identify the hot spot amino acid residues. Consensus-based amino acid substitution was applied to evaluate multiple sequence alignment of homologs and identified 20 amino acids positions by following Jensen-Shnnon Divergence method. 3D models of 20 selected mutants were analyzed for conformational transition in protein structures by using NMSim server. Two selected mutants T6K and I17M of Xyl-11A retained 40, 60% residual activity respectively, at 85 °C for 120 min as compared to wild type enzyme which retained 37% initial activity under same conditions, confirming the enhanced thermostability of mutants. The present study showed a good approach for the identification of promising amino acid residues responsible for enhancing the thermostability of enzymes of industrial importance.