This report describes an approach to recover SARS-CoV-2 RNA from rapid antigen test kit (RTK-antigen) cassettes for whole-genome sequencing (WGS). RNA samples were recovered from 33 RTK-antigen cassettes for WGS, with 18 samples achieving more than 80% genome coverage. This work provides a proof-of-concept that positive RTK-antigen cassettes can be safely transported, stored and subjected to WGS, enabling swift identification of circulating variants.

Multiple morphological abnormalities of the flagella (MMAF) represent a severe form of sperm defects leading to asthenozoospermia and male infertility. In this study, we identified a novel homozygous splicing mutation (c.871-4 ACA>A) in the adenylate kinase 7 (AK7) gene by whole-exome sequencing in infertile individuals. Spermatozoa from affected individuals exhibited typical MMAF characteristics, including coiled, bent, short, absent, and irregular flagella. Transmission electron microscopy analysis showed disorganized axonemal structure and abnormal mitochondrial sheets in sperm flagella. Immunofluorescence staining confirmed the absence of AK7 protein from the patients' spermatozoa, validating the pathogenic nature of the mutation. This study provides direct evidence linking the AK7 gene to MMAF-associated asthenozoospermia in humans, expanding the mutational spectrum of AK7 and enhancing our understanding of the genetic basis of male infertility.
Humans ; Male ; Sperm Tail/ultrastructure* ; Homozygote ; Consanguinity ; Asthenozoospermia/pathology* ; Infertility, Male/genetics* ; Mutation ; Pakistan ; Adenylate Kinase/genetics* ; Adult ; Pedigree ; RNA Splicing ; Exome Sequencing ; Spermatozoa

Male infertility can result from impaired sperm motility caused by multiple morphological abnormalities of the flagella (MMAF). Distinct projections encircling the central microtubules of the spermatozoal axoneme play pivotal roles in flagellar bending and spermatozoal movement. Mammalian sperm-associated antigen 17 ( SPAG17 ) encodes a conserved axonemal protein of cilia and flagella, forming part of the C1a projection of the central apparatus, with functions related to ciliary/flagellar motility, skeletal growth, and male fertility. This study investigated two novel homozygous SPAG17 mutations (M1: NM_206996.2, c.829+1G>T, p.Asp212_Glu276del; and M2: c.2120del, p.Leu707*) identified in four infertile patients from two consanguineous Pakistani families. These patients displayed the MMAF phenotype confirmed by Papanicolaou staining and scanning electron microscopy assays of spermatozoa. Quantitative real-time polymerase chain reaction (PCR) of patients' spermatozoa also revealed a significant decrease in SPAG17 mRNA expression, and immunofluorescence staining showed the absence of SPAG17 protein signals along the flagella. However, no apparent ciliary-related symptoms or skeletal malformations were observed in the chest X-rays of any of the patients. Transmission electron microscopy of axoneme cross-sections from the patients showed incomplete C1a projection and a higher frequency of missing microtubule doublets 1 and 9 compared with those from fertile controls. Immunofluorescence staining and Western blot analyses of spermatogenesis-associated protein 17 (SPATA17), a component of the C1a projection, and sperm-associated antigen 6 (SPAG6), a marker of the spring layer, revealed disrupted expression of both proteins in the patients' spermatozoa. Altogether, these findings demonstrated that SPAG17 maintains the integrity of spermatozoal flagellar axoneme, expanding the phenotypic spectrum of SPAG17 mutations in humans.
Humans ; Male ; Infertility, Male/pathology* ; Sperm Tail/ultrastructure* ; Homozygote ; Microtubule-Associated Proteins/genetics* ; Axoneme/genetics* ; Spermatozoa/ultrastructure* ; Adult ; Mutation ; Sperm Motility/genetics* ; Pedigree ; Microtubules ; Microtubule Proteins/genetics*

The syndrome of multiple morphological abnormalities of the sperm flagella (MMAF) is one of the most serious kinds of sperm defects, leading to asthenoteratozoospermia and male infertility. In this study, we use whole-exome sequencing to identify genetic factors that account for male infertility in a patient born from a consanguineous Pakistani couple. A homozygous frameshift mutation (c.1399_1402del; p.Gln468ArgfsTer2) in axonemal dynein light chain domain containing 1 ( AXDND1 ) was identified in the patient. Sanger sequencing data showed that the mutation was cosegregated recessively with male infertility in this family. Papanicolaou staining and scanning electron microscopy analysis of the sperm revealed severely abnormal flagellar morphology in the patient. Immunofluorescence and western blot showed undetectable AXDND1 expression in the sperm of the patient. Transmission electron microscopy analysis showed disorganized sperm axonemal structure in the patient, particularly missing the central pair of microtubules. Immunofluorescence staining showed the absence of sperm-associated antigen 6 (SPAG6) and dynein axonemal light intermediate chain 1 (DNALI1) signals in the sperm flagella of the patient. These findings indicate that AXDND1 is essential for the organization of flagellar axoneme and provide direct evidence that AXDND1 is a MMAF gene in humans, thus expanding the phenotypic spectrum of AXDND1 frameshift mutations.
Humans ; Male ; Sperm Tail/ultrastructure* ; Frameshift Mutation ; Infertility, Male/pathology* ; Pakistan ; Pedigree ; Consanguinity ; Axonemal Dyneins/genetics* ; Adult ; Spermatozoa ; Exome Sequencing

Introduction: Assessing radiation exposure is a vital step in determining the potential health risks associated with radiation and identifying the necessity for protective measures. The study assesses the occupational radiation dose encountered by undergraduate students specializing in diagnostic imaging and radiotherapy (DIR) during their clinical training in the radiology department. Materials and methods: An analysis was conducted to compare individual exposure doses with the dose limits established by the International Commission on Radiological Protection (ICRP), concurrently assessing the variations in mean effective doses among distinct student cohorts. Optically stimulated luminescence dosimeters (OSLD) were employed to monitor 143 students across five cohorts from 2017 to 2022. The electronic Secondary Standard Dosimetry Laboratory (e-SSDL) information security management system tracked the deep (Hp(10)) and shallow (Hp(0.07)) dose equivalents of DIR students. Results: The mean values for accumulated Hp(10) and Hp(0.07) were found to be 0.51 and 0.50 mSv, respectively, averaged over 800 hours of clinical training. The highest recorded exposure was 1.30 mSv, signifying that the radiation exposure experienced by DIR students was well below the threshold. The mean annual effective radiation dose per student amounted to 0.17 mSv. Small but significant difference (p < 0.001) in the radiation exposure between five cohorts was observed with increments over a 5-year period. Conclusion: The students' radiation dose was notably below the ICRP dose limit of 6 mSv/ year designated for trainees. These results indicate the efficacy of the existing radiation protection measures during clinical training, reducing the likelihood of overexposure

Malaria is a life-threatening, preventable, and curable vector borne disease caused by parasites that are transmitted to people through the bites of infected female Anopheles. The WHO Global Report 2010-2016 reported insecticide resistance in malaria. The main objective of this study is to determine the effectiveness of new generation Long-Lasting Insecticidal Nets (LLIN) compared to standard LLIN and untreated nets in terms of the mortality rate of adult female Anopheles gambiae. A comprehensive review of the literature was published in three databases (PubMed, Ovid, EBSCO Host) since 2010. Publications were searched with keywords including malaria, long-lasting treated bed net, long lasting insecticide-treated bed net, LLIN, and experimental hut. The search has identified 60 articles. Based on the PRISMA flowchart, 10 articles are qualified for data collection and analysis. The gathered data was analysed using Review Manager. Following meta-analysis between subgroups, a risk difference of 0.31 between standard LLINs versus untreated net (p<0.001, I²=100% 95% CI:0.01,0.60). A comparison of upgraded LLINs with the untreated net has shown a significant difference with a pooled risk difference of 0.54 favours upgraded LLINs (p<0.001, I²=100% 95% CI: 0.54,0.84). Comparison between upgraded LLINs versus standard gave an overall risk difference of 0.24 (p < 0.001, I² = 100%, 95% CI: 0.10–0.39). Upgraded LLINs significantly increase Anophelesmortality compared to standard LLINs and untreated nets, suggesting their potential for improved malaria control. Thus, using upgraded nets in the field and translating them into malaria preventive programs would help achieve the target and improve healthoutcomes for those living in endemic areas.

Dengue remains a public health threat that consumes a significant number of resources for its prevention and control. This systematic review aimed to solidify recent costing evidence in dengue management among South East Asian (SEA) countries. All studies conducted between 2010 and 2020 were retrieved using four international databases i.e. PubMed, Scopus, Web of Science, and Emerald Insight. The review was reported according to PRISMA guidelines. Quality assessments were done independently by two reviewers using a checklist adapted for the cost of illness studies. We identified 13 original articles representing several SEA countries. Among the common reported costing measure include total cost/ health expenditure; direct medical cost; direct non-medical cost; and indirect cost. The estimated total cost for dengue management varied between countries largely due to the difference in the total incidence of dengue cases. The estimated cost spent on dengue per capita GDP ranges from less than 0.001% to 0.1%, depending on the recorded number of dengue cases of the year. The majority of the articles focused on the economic burden from the perspective of treatment such as hospitalization and ambulatory care. In a nutshell, the economic burden of managing dengue infection is costly and the evidence suggests a steady increase in health expenditure with the growing number of dengue cases

It is paramount to assess the risk of biases in may arise from diagnostic test accuracy (DTA) study as it will affect the accuracy and validity of the tests. These biases can be found in published researches and here we look at COVID-19 DTA studies. The evaluation of bias risk in diagnostic research is mainly performed using QUADAS-2. The aim of this review was to determine potential selection and information biases in diagnostic test accuracy studies and strategies to minimize risk of biases. Literature review related to diagnostic test accuracy study is identified through an online search of databases namely PubMed, ScienceDirect, Research Gate, Google Scholar, and official government websites range. Six potential biases in four QUADAS-2 domains are identified in COVID-19 diagnostic test accuracy study which are 1) spectrum bias in patient selection; 2) interpretation bias in index test; 3) differential misclassification bias and nondifferential misclassification bias in reference standard; and 4) partial verification bias and differential verification bias in patient flow. The identified biases exert effects on accuracy of COVID-19 diagnostic tests. Six strategies are recommended to reduce these biases, hence, improving the accuracy of COVID-19 diagnostic tests. The best diagnostic test can give benefits to the population in the mass screening program during COVID-19.

Severe hyponatraemia is defined as a sodium level of less than 120 mEq/L, and it is frequently accompanied by neurological symptoms like coma, convulsions, respiratory arrest, and death. Clinical cardiac toxicity from hyponatremia, such as bradyarrhythmia, is extremely rare. In this article, we present a case of acute severe hyponatraemia that induced unstable bradyarrhythmia and led to refractory bradycardia, which did not improve despite receiving treatment in accordance with the standard Advanced Cardiovascular Life Support protocol. The patient’s bradyarrhythmia has completely resolved with the administration of 3% hypertonic saline, which restored her sodium levels. Due to the possibility that severe hyponatremia may contribute to the aetiology of cardiac malfunction, this case raises awareness about the significance of closely monitoring electrocardiograms and telemetry in patients with severe hyponatremia.

Aims: The study was aimed to explore the antimicrobial potential of ethanolic leaf extracts of Eucalyptus globulus, Moringa oliefera, Syzygium cumini and Citrus limon against antibiotic-resistant Clostridium perfringens type D (n=5). Methodology and results: Antibiotic resistance pattern of C. perfringens type D isolates against tetracycline, gentamicin, ceftriaxone, amoxicillin and streptomycin was evaluated by disc diffusion method. Well diffusion and micro broth dilution methods were used to determine the anti-bacterial activity, sub-inhibitory concentrations and antibiotic resistance modulating effects of the plant extracts. Ethanolic extract of E. globules was selected to evaluate its modulatory impact and subjected to GC-MS analysis to separate and identify the phytochemicals. The results showed that the isolates were resistant to gentamicin (0 ± 0.00 mm), streptomycin (0 ± 0.00 mm), tetracycline (13.2 ± 2.28 mm) and ceftriaxone (0 ± 0.00 mm) while sensitive to amoxicillin (23.8 ± 1.30 mm) and tetracycline (13.2 ± 2.28 mm). Eucalyptus globulus exhibited the maximum anti-bacterial activity with a zone of inhibition (ZOI) of 14.6 ± 0.54 mm and minimum inhibitory concentration (MIC) (1500 ± 947.85 µg/mL). Other plant extracts (M. oliefera, S. cumini and C. limon) also showed anti-bacterial activity but couldn’t modulate the resistance. The activity of ceftriaxone associated with E. globulus extract was improved with 20.2 ± 0.20 mm ZOI at 78.125 µg/mL sub-inhibitory concentration. Conclusion, significance and impact of study: The study results indicate the possible use of the ethanolic extract of E. globulus alone or in combination with common antibiotics for the treatment of C. perfringens infections in small ruminants.