Molecular characterization of rotavirus indicates predominance of G9P[4] genotype among children with acute gastroenteritis: First report after vaccine introduction in Pakistan.

Globally, Group A rotavirus (RVA) is the leading cause of acute gastroenteritis in children under 5 years old, with Pakistan having the highest rates of RVA-related morbidity and mortality. The current study aims to determine the genetic diversity of rotavirus and evaluate the impact of Rotarix-vaccine introduction on disease epidemiology in Pakistan. A total of 4749 children, hospitalized with acute gastroenteritis between 2018 and 2020, were tested at four hospitals in Lahore and Karachi. Of the total, 19.3% (918/4749) cases were tested positive for RVA antigen, with the positivity rate varying annually (2018 = 22.7%, 2019 = 14.4%, 2020 = 20.9%). Among RVA-positive children, 66.3% were under 1 year of age. Genotyping of 662 enzyme-linked immuno sorbent assay-positive samples revealed the predominant genotype as G9P[4] (21.4%), followed by G1P[8] (18.9%), G3P[8] (11.4%), G12P[6] (8.7%), G2P[4] (5.7%), G2P[6] (4.8%), and 10.8% had mixed genotypes. Among vaccinated children, genotypes G9P[4] and G12P[6] were more frequently detected, whereas a decline in G2P[4] was observed. Phylogenetic analysis confirmed the continued circulation of indigenous genotypes detected earlier in the country except G9 and P[6] strains. Our findings highlight the predominance of G9P[4] genotype after the vaccine introduction thus emphasizing continual surveillance to monitor the disease burden, viral diversity, and their impact on control of rotavirus gastroenteritis in children.

Municipal wastewater treatment with corrugated PVC carrier anaerobic baffled reactor.

The anaerobic baffled reactor (ABR) is a promising solution for decentralized wastewater treatment due to its low operation cost as compared to the activated sludge process, but it requires comparatively higher hydraulic retention time (HRT). This ultimately increases land requirement, capital and construction cost of treatment plant. This study investigates performance of ABR using polyvinyl chloride (PVC) corrugated pipe as carrier media to improve biomass retention capacity and treatment performance of reactor with the aim to reduce HRT. Comparative performance of two ABRs with and without carrier media was analyzed under mesophilic conditions (35 ± 1 °C) for organics and total suspended solids (TSS) removal at HRTs of 24, 18, 12, 8, 6 and 4 h. Results showed that at HRTs of 24-08 h, the organics removal performance of the carrier anaerobic baffled reactor (CABR) was better than ABR and was in the range of 77-81% for CABR as compared to 64-70% for ABR. However, on further decrease in HRT to 6 h, CABR sustained the treatment with organics removal of 80%, while ABR performance reduced to 58%, creating a performance difference of 38%. Average total suspended solids (TSS) removal was in the range of 76-83% at all HRTs for both reactors. Therefore, this study identified CABR with PVC carrier media as an effective low-HRT reactor for organics and SS removal with less land area requirement.

Radiographic patterns on Chest X-ray as a supporting imaging tool in triaging of suspected Corona Virus Disease (COVID) patients.

Objective: To evaluate the radiographic patterns on Chest X-Ray (CXR) in accordance with Modified Brixia Scoring as supporting imaging tool in triaging of Corona Virus Disease (COVID-19) pneumonia. Methods: In this cross-sectional study, chest radiographs of suspected COVID patients at emergency triage, Lady Reading Hospital (LRH) from April 18th to July 22nd 2020 were evaluated for patterns of COVID pneumonia and scored in accordance with modified Brixia score. Each zone was categorized as score of "one" for interstitial pattern, "two" for mixed interstitial /alveolar pattern and "three" for alveolar pattern. Radiographic patterns consistent with COVID pneumonia or patients having strong clinical suspicion were advised Polymerase Chain Reaction (PCR) tests. Results: Total of 2,225 individuals were screened for patterns of COVID-19 pneumonia on chest radiograph. Out of these 1465(65.8%) had normal chest radiograph and 760(34.2%) had abnormal findings. Out of the total, 648 suspected COVID patients were selected for PCR. The radiographic patterns ranged from mixed interstitial/alveolar pattern in 261(40.3%) patients, alveolar pattern in 231(35.6%), interstitial pattern in 87(13.4%), pleural effusion in 12(1.9%), other findings in 5(0.8%) while 52(8%) suspected Covid patients had normal radiographs. The PCR was positive in 326(50.3%), negative in 100(15.4%) and inconclusive in 60(9.3%) while 162(25%) were lost to follow up. Amongst the 52 suspected Covid patients having normal chest radiographs, 10 were positive on PCR, 21 negative, seven suspected and two inconclusive, while 12 were lost to follow up. Conclusion: Chest radiograph is used for triaging of suspected COVID pneumonia patients in emergency settings. It assesses the severity of disease according to modified Brixia scoring for treatment plan.

Helicobacter pylori adhesins: HpaA a potential antigen in experimental vaccines for H. pylori.

BACKGROUND: Helicobacter pylori is a gram-negative bacterium involved in many gastric pathologies such as ulcers and cancers. Although the treatment for this infection has existed for several years, the development of a vaccine is nevertheless necessary to reduce the severe forms of the disease. For more than three decades, many advances have been made particularly in the understanding of virulence factors as well as the pathogenesis of gastric diseases caused by H. pylori. Among these key virulence factors, specific antigens have been identified: Urease, Vacuolating cytotoxin A (VacA), Cytotoxin-associated gene A (CagA), Blood group antigen-binding adhesin (BabA), H. pylori adhesin A (HpaA), and others. OBJECTIVES: This review will focus on H. pylori adhesins, in particular, on HpaA and on the current knowledge of H. pylori vaccines. METHODS: All of the information included in this review was retrieved from published studies on H. pylori adhesins in H. pylori infections. RESULTS: These proteins, used in their native or recombinant forms, induce protection against H. pylori in experimental animal models. CONCLUSION: H. pylori adhesins are known to be promising candidate vaccines against H. pylori. Future research should be carried out on adhesins, in particular, on HpaA.

Standardized approach to result analysis and interpretation of initial experience of coronary computed tomography angiography in a tertiary care hospital.

OBJECTIVE: To have a systematic standardized approach to performing and interpretation of coronary CT Angiography (CTA) in order to maintain and enhance the diagnostic accuracy of the imaging modality. METHODS: This retrospective observational study was performed in Radiology Department, Lady Reading Hospital, Medical Teaching Institute, Peshawar, Pakistan from August 2018 to September 2019. Patients referred for coronary CT angiography were screened and prepared in radiology department. The examination was performed on Toshiba-160 slice CT Scanner (Prime Aquilion) utilizing standardized protocols tailored towards optimum image acquisition. Interpretation of the images were based on the guidelines provided by the Society of Cardiovascular Computed Tomography (SCCT). RESULTS: Total 95 CTCA procedures were performed in the department, out of which 85 were included in study showing 49(57%) as normal and 36(42%) were positive for coronary disease. Of the abnormal cases 16(18.8%) had mild disease, nine (10.6%) patients had moderate disease, while severe disease was noted in 11 (12.9%) cases. Coronary quantitative stenosis revealed five cases (5.9%) with LAD involvement only and five (5.9%) with severe triple vessel disease. The remaining varied in degree of stenosis and number of segments involved. Post CABG were 14 cases (16%) and native arteries showed triple vessel disease. CTA for percutaneous stent patency were three (3.5%) cases with 100% stent patency. CONCLUSION: Patient selection with tailored protocols are the mainstay for achieving optimal images. This form the basis for accurate interpretation, based on a standardized and systematic approach, utilizing various post processing tools, in order to maintain the high diagnostic accuracy of this semi-invasive, safe imaging modality in a variety of patients suspected of coronary artery disease, coronary artery bypass grafts and stent patency.

Analysis of molecular subtypes and antibiotic resistance in Treponema pallidum isolates from blood donors in Khyber Pakhtunkhwa, Pakistan.

Syphilis, caused by Treponema pallidum, is resurging globally. Molecular typing allows for the investigation of its epidemiology. In Pakistan and other nations, T. pallidum subsp. pallidum has developed widespread macrolide resistance in the past decade. A study at the Peshawar Regional Blood Centre from June 2020-June 2021 analyzed serum samples from 32,812 blood donors in Khyber Pakhtunkhwa, Pakistan, to assess circulating T. pallidum strains and antibiotic resistance. Blood samples were initially screened for T. pallidum antibodies using a chemiluminescent microparticle immunoassay (CMIA). CMIA-reactive samples underwent polymerase chain reaction (PCR) targeted the polA, tpp47, bmp, and tp0319 genes. PCR-positive samples were further analyzed for molecular subtyping using a CDC-developed procedure and tp0548 gene examination. All PCR-positive samples were analyzed for the presence of point mutations A2058G and A2059G in 23S rRNA, as well as the G1058C mutation in 16S rRNA. These mutations are known to impart antimicrobial resistance to macrolides and doxycycline, respectively. Out of 32,812 serum samples, 272 (0.83%) were CMIA-reactive, with 46 being PCR-positive. Nine T. pallidum subtypes were identified, predominantly 14d/f. The A2058G mutation in 23S rRNA was found in 78% of cases, while G1058C in 16S rRNA and A2059G in 23S rRNA were absent. The research found donor blood useful for assessing T. pallidum molecular subtypes and antibiotic resistance, especially when chancres are not present. The prevalent subtype was 14d/f (51.85%), and the high macrolide resistance of 36 (78%) indicates caution in using macrolides for syphilis treatment in Khyber Pakhtunkhwa, Pakistan.

Insights into climate change dynamics: A tourism climate index-based evaluation of Gilgit-Baltistan, Pakistan.

The tourism industry is a significant contribution to the economy of many countries, including Pakistan. However, its activities often have a negative impact on the environment, particularly related to climate change. Notably, Pakistan ranks fifth among countries most affected by climate change, which requires a targeted analysis of the tourism sector to determine its potential impacts. Despite the critical nature of this issue, there is currently a lack of research that examines how climate change specifically impacts Pakistan's tourism industry. This study aims to address this gap by using the Tourism Climate Index (TCI) to assess the impact of climate change on the suitability of different tourism locations across the country. Our results show that popular tourist destinations such as Gilgit-Baltistan experience their peak season in the warmer third quarter (summer), which corresponds to the highest observed TCI values. This suggests that warmer temperatures could boost tourism activity and spending in these regions. Furthermore, our analysis shows that air temperature plays a crucial role in determining tourist comfort levels and significantly influences tourists' TCI values and sensory experiences. The results of this study show that the TCI methodology can be an effective tool for further research into the geographical impacts of climate change on tourism. By identifying necessary changes due to climatic changes, future studies could provide valuable insights into how the tourism industry can adapt to and reduce its environmental footprint.

A novel homozygous splicing mutation in AK7 causes multiple morphological abnormalities of sperm flagella in patients from consanguineous Pakistani families.

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.

Combined clinical accuracy of inflammatory markers and ultrasound for the diagnosis of acute appendicitis.

Introduction: Acute appendicitis is traditionally a clinical diagnosis where there are a range of diagnostic accuracies reported. The role of ultrasound is to improve specificity and decrease negative appendicectomy rate. It is a simple, non-invasive, easily available technique.The purpose of this study is to determine the diagnostic accuracy of an ultrasound in combination with total leukocyte count, neutrophil percentage and C-reactive protein in diagnosing acute appendicitis. Methods: This study includes consecutive sampling of suspected patients from January 2021 to February 2022 with the approval of the ethical and research committee. Clinical and personal demographics and characteristics of patients were collected, including age, gender, symptoms and clinical signs. Ultrasonographic findings of fluid-filled appendiceal diameter of more than 6 mm, periappendiceal echogenic mesentry and an appendicolith were primary positive features. Laboratory inflammatory markers of total leukocyte count, neutrophil percentage and C-reactive protein were also included. Results: A total of 250 patients were included with a mean age of 25 ± 9.79 years. Total leukocyte count showed the highest sensitivity (77.68%), followed by neutrophil percentage (69.96%), C-reactive protein (67.10%) and ultrasound (62.96%). While ultrasound had the best specificity (70.59%), it was followed by C-reactive protein and total leukocyte count (64.71%) and neutrophil percentage (58.82%), respectively. The sensitivity and specificity (99% and 98%) increased significantly when all four tests were combined. Conclusions: Clinical assessment with laboratory inflammatory markers and ultrasound improves the early diagnosis of appendicitis and decreases the false-positive appendicitis diagnosis, hence saving surgeons' time and relieving patients from unnecessary appendicectomies.

Novel Mutations in MPT64 Secretory Protein of Mycobacterium tuberculosis Complex.

Tuberculosis (TB) is a global health problem caused by the Mycobacterium tuberculosis complex (MTBC). These bacteria secrete various proteins involved in the pathogenesis and persistence of MTBC. Among the secretory proteins, MPT64 (Rv1980C) is highly conserved and is also known as a major culture filtrate that is used in rapid diagnosis of MTBC. In the current study, we aimed to find the mutation in this highly conserved protein in isolates from the Pashtun-dominant province of Pakistan. We analyzed 470 M. tuberculosis whole-genome sequences of Khyber Pakhtunkhwa Province. Mutations in the MPT64 gene were screened through TB-Profiler and BioEdit software tools. The DynaMut web server was used to analyze the impact of the mutation on protein dynamics and stability. Among 470 MTB genomes, three non-synonymous mutations were detected in nine isolates, and one synonymous mutation (G208A) was found in four isolates. Mutation G211T (F159L), which was detected at the C-terminal domain of the protein in six isolates, was the most prominent. The second novel mutation, T480C (I70V), was detected in two isolates at the C-terminal side of the protein structure. The third novel mutation, A491C (L66R), was detected in a single isolate at the N-terminal side of the MPT64 protein. The effect of these three mutations was destabilizing on the protein structure. The molecular flexibility of the first two mutations increased, and the last one decreased. MPT64 is a highly conserved secretory protein, harboring only a few mutations. This study provides useful information for better managing the diagnosis of MTB isolates in high TB-burden countries.

The Impact of Urbanization on Urban Heat Island: Predictive Approach Using Google Earth Engine and CA-Markov Modelling (2005-2050) of Tianjin City, China.

Urbanization has adverse environmental effects, such as rising surface temperatures. This study analyzes the relationship between the urban heat island (UHI) intensity and Tianjin city's land cover characteristics. The land use cover change (LUCC) effects on the green areas and the land surface temperature (LST) were also studied. The land cover characteristics were divided into five categories: a built-up area, an agricultural area, a bare area, a forest, and water. The LST was calculated using the thermal bands of spatial images taken from 2005 to 2020. The increase in the built-up area was mainly caused by the agricultural area decreasing by 11.90%. The average land surface temperature of the study area increased from 23.50 to 36.51 °C, and the region moved to a high temperature that the built-up area's temperature increased by 1.5%. Still, the increase in vegetation cover was negative. From 2020 to 2050, the land surface temperature is expected to increase by 9.5 °C. The high-temperature areas moved into an aerial distribution, and the direction of urbanization determined their path. Urban heat island mitigation is best achieved through forests and water, and managers of urban areas should avoid developing bare land since they may suffer from degradation. The increase in the land surface temperature caused by the land cover change proves that the site is becoming more urbanized. The findings of this study provide valuable information on the various aspects of urbanization in Tianjin and other regions. In addition, future research should look into the public health issues associated with rapid urbanization.

Novel mutations in genes of the IL-12/IFN-γ axis cause susceptibility to tuberculosis.

BACKGROUND: The IL-12/23/ISG15-IFN-γ pathway is the main immunological pathway for controlling intra-macrophagic microorganisms such as Mycobacteria, Salmonella, and Leishmania spp. Consequently, upon mutations in genes of the IL-12/23/ISG15-IFN-γ pathway cause increased susceptibility to intra-macrophagic pathogens, particularly to Mycobacteria. Therefore, the purpose of this study was to characterize the mutations in genes of the IL-12/23/ISG15-IFN-γ pathway in severe tuberculosis (TB) patients. METHODS: Clinically suspected TB was initially confirmed in four patients (P) (P1, P2, P3, and P4) using the GeneXpert MTB/RIF and culturing techniques. The patients' Peripheral blood mononuclear cells (PBMCs) were then subjected to ELISA to measure Interleukin 12 (IL-12) and interferon gamma (IFN-γ). Flow cytometry was used to detect the surface expressions of IFN-γR1 and IFN-γR2 as well as IL-12Rß1and IL-12Rß2 on monocytes and T lymphocytes, respectively.The phosphorylation of signal transducer and activator of transcription 1(STAT1) on monocytes and STAT4 on T lymphocytes were also detected by flow cytometry. Sanger sequencing was used to identify mutations in the IL-12Rß1, STAT1, NEMO, and CYBB genes. RESULTS: P1's PBMCs exhibited reduced IFN-γ production, while P2's and P3's PBMCs exhibited impaired IL-12 induction. Low IL-12Rß1 surface expression and reduced STAT4 phosphorylation were demonstrated by P1's T lymphocytes, while impaired STAT1 phosphorylation was detected in P2's monocytes. The impaired IκB-α degradation and abolished H2O2 production in monocytes and neutrophils of P3 and P4 were observed, respectively. Sanger sequencing revealed novel nonsense homozygous mutation: c.191 G>A/p.W64 * in exon 3 of the IL-12Rß1 gene in P1, novel missense homozygous mutation: c.107 A>T/p.Q36L in exon 3 of the STAT1 gene in P2, missense hemizygous mutation:: c.950 A>C/p.Q317P in exon 8 of the NEMO gene in P3, and nonsense hemizygous mutation: c.868 C>T/p.R290X in exon 8 of CYBB gene in P4. CONCLUSION: Our findings broaden the clinical and genetic spectra associated with IL-12/23/ISG15-IFN-γ axis anomalies. Additionally, our data suggest that TB patients in Pakistan should be investigated for potential genetic defects due to high prevalence of parental consanguinity and increased incidence of TB in the country.

Biallelic mutations in RNA-binding protein ADAD2 cause spermiogenic failure and non-obstructive azoospermia in humans.

STUDY QUESTION: What are some pathogenic mutations for non-obstructive azoospermia (NOA) and their effects on spermatogenesis? SUMMARY ANSWER: Biallelic missense and frameshift mutations in ADAD2 disrupt the differentiation of round spermatids to spermatozoa causing azoospermia in humans and mice. WHAT IS KNOWN ALREADY: NOA is the most severe cause of male infertility characterized by an absence of sperm in the ejaculate due to impairment of spermatogenesis. In mice, the lack of the RNA-binding protein ADAD2 leads to a complete absence of sperm in epididymides due to failure of spemiogenesis, but the spermatogenic effects of ADAD2 mutations in human NOA-associated infertility require functional verification. STUDY DESIGN SIZE DURATION: Six infertile male patients from three unrelated families were diagnosed with NOA at local hospitals in Pakistan based on infertility history, sex hormone levels, two semen analyses and scrotal ultrasound. Testicular biopsies were performed in two of the six patients. Adad2 mutant mice (Adad2Mut/Mut) carrying mutations similar to those found in NOA patients were generated using the CRISPR/Cas9 genome editing tool. Reproductive phenotypes of Adad2Mut/Mut mice were verified at 2 months of age. Round spermatids from the littermates of wild-type (WT) and Adad2Mut/Mut mice were randomly selected and injected into stimulated WT oocytes. This round spermatid injection (ROSI) procedure was conducted with three biological replicates and >400 ROSI-derived zygotes were evaluated. The fertility of the ROSI-derived progeny was evaluated for three months in four Adad2WT/Mut male mice and six Adad2WT/Mut female mice. A total of 120 Adad2Mut/Mut, Adad2WT/Mut, and WT mice were used in this study. The entire study was conducted over 3 years. PARTICIPANTS/MATERIALS SETTING METHODS: Whole-exome sequencing was performed to detect potentially pathogenic mutations in the six NOA-affected patients. The pathogenicity of the identified ADAD2 mutations was assessed and validated in human testicular tissues and in mouse models recapitulating the mutations in the NOA patients using quantitative PCR, western blotting, hematoxylin-eosin staining, Periodic acid-Schiff staining, and immunofluorescence. Round spermatids of WT and Adad2Mut/Mut mice were collected by fluorescence-activated cell sorting and injected into stimulated WT oocytes. The development of ROSI-derived offspring was evaluated in the embryonic and postnatal stages. MAIN RESULTS AND THE ROLE OF CHANCE: Three recessive mutations were identified in ADAD2 (MT1: c.G829T, p.G277C; MT2: c.G1192A, p.D398N; MT3: c.917_918del, p.Q306Rfs*43) in patients from three unrelated Pakistani families. MT1 and MT2 dramatically reduced the testicular expression of ADAD2, likely causing spermiogenesis failure in the NOA patients. Immunofluorescence analysis of the Adad2Mut/Mut male mice with the corresponding MT3 mutation showed instability and premature degradation of the ADAD2 protein, resulting in the spermiogenesis deficiency phenotype. Through ROSI, the Adad2Mut/Mut mice could produce pups with comparable embryonic development (46.7% in Adad2Mut/Mut versus 50% in WT) and birth rates (21.45 ± 10.43% in Adad2Mut/Mut versus 27.5 ± 3.536% in WT, P = 0.5044) to WT mice. The Adad2WT/Mut progeny from ROSI (17 pups in total via three ROSI replicates) did not show overt developmental defects and had normal fertility. LARGE SCALE DATA: N/A. LIMITATIONS REASONS FOR CAUTION: This is a preliminary report suggesting that ROSI can be an effective treatment for infertile Adad2Mut/Mut mice. Further assisted reproductive attempts need to be carefully examined in humans during clinical trials. WIDER IMPLICATIONS OF THE FINDINGS: Our work provides functional evidence that mutations in the ADAD2 gene are deleterious and cause consistent spermiogenic defects in both humans and mice. In addition, preliminary results show that ROSI can help Adad2Mut/Mut to produce biological progeny. These findings provide valuable clues for genetic counselling on the ADAD2 mutants-associated infertility in human males. STUDY FUNDING/COMPETING INTERESTS: This work was supported by the National Natural Science Foundation of China (32000587, U21A20204, and 32061143006), and the National Key Research and Developmental Program of China (2019YFA0802600 and 2021YFC2700202). This work was also supported by Institute of Health and Medicine, Hefei Comprehensive National Science Center, Hefei, China. The authors declare no competing interests.

Morpho-anatomical characterization of some members of euglenophycota (algae) of north-east Punjab, Pakistan based on light microscopy (LM) and scanning electron microscopy (SEM).

The study was carried out in four districts, that is, Gujranwala, Gujarat, Narowal and Sialkot of Punjab, Pakistan. The sampling was carried out randomly in different seasons from the water bodies especially from wastewater. Twenty-one species belonging to Euglenophycota were identified using light microscopy and scanning electron microscopy from which 04 species belong to genus Phacus, 02 species belonging to Trachelomonas and Euglena based on light microscopy. It was observed that Euglena was the most diverse genus and it is supposed to be the indicator species of the polluted water. It was observed that E. oblonga was found in maximum pH range, that is, 7.0-11.0. Similarly, E. brevicaudatus was found in maximum EC, that is, 169 ± 1.5 ms/cm these outcomes indicated that for internal examination along with LM, SEM was necessary for correct identification of algal sample up to specie level.

Morpho-anatomical characters of family Oscillatoriaceae (algae) identified using primitive (LM) and advance technique (SEM) from Punjab, Pakistan.

In this study, 34 species belonging to the class Nostocophyceae order Nostocales and family Oscillatoriaceae were identified based on light microscopy (LM) and scanning electron microscopy (SEM). All species belong to nine genera of which two species belong to Arthrospira, six species to Lyngbya, ten species belongs to Oscillatoria, six species belong to Spirulina, seven species to Phormidium, three species to Microcoleus. The morpho-anatomical characters were identified among different species using LM and SEM. These studies highlighted that north-eastern area of Punjab was blessed with the diversity of algae. These outcomes also indicated that for internal examination along with LM, SEM was necessary for correct identification of algal sample up to specie level. RESEARCH HIGHLIGHTS: The different Oscillatoria species abundance in waste-water of four different areas. The sampling was carried out randomly in different seasons from the water bodies especially from waste-water. The morpho-anatomical characters were recognized using light microscopy (LM) and scanning electron microscopy (SEM). Thirty-four species belonging to the class Nostocophyceae order Nostocales and family Oscillatoriaceae were identified based on LM and SEM.

Light and scanning electron microscopy of Datura stramonium L. extract and its biological applications.

The study deals with the LM and SEM (light microscopy and scanning electron microscopy) of Datura stramonium L. extract and to evaluate its phytochemical, antimicrobial, and antioxidant activities. The plant sample collected from different Lahore region was dried under shade. The plant extract was prepared using different solvents (n-hexane, chloroform, ethyl acetate, and methanol). Phytochemical screening performed qualitatively for alkaloids, carbohydrates, flavonoids, saponins, and tannins. Radical scavenging potential of plant extracts was estimated and ethyl acetate extract exhibited maximum percentage scavenging ability of 92.2 ± 0.09. Total antioxidant capacity of different extracts was measured by phosphomolybdate method. The ethyl acetate and methanolic extracts showed maximum values of 72.3 ± 0.08% and 79 ± 0.15% with respect to gallic acid. Total phenolic contents were determined through Folin-Ciocalteau method and highest value of 495.2 ± 3.4 was obtained by methanolic extract. DPPH-TLC autography was used for the visual detection of antioxidant compounds in different plant extracts. Antibacterial activity of plant extract was measured by disc diffusion method against microorganisms named Bacillus licheniformis, Bacillus subtilis (Gram positive), and Salmonella typhimurium, and Proteus mirabilis (Gram negative). Because of the vast medicinal importance of D. stramonium L. it would be beneficial to study LM and SEM for correct identification of plant and to determine phytochemicals. RESEARCH HIGHLIGHTS: LM and SEM (light microscopy and scanning electron microscopy) of Datura stramonium L. was studied for the exact identification of plant. To evaluate the biological activities of pharmaceutically important plant. Antioxidant and antimicrobial activity were determined. D. stramonium L. proved to be medicinally important plant.

Gastronomic Tourism and Tourist Motivation: Exploring Northern Areas of Pakistan.

Gastronomic tourism is becoming an essential consideration among tourists when choosing a tourist destination. One of the main reasons for visiting a specific destination for almost 15% of tourists is "gastronomy". This has led to the development of a new kind of tourism called "Gastronomic Tourism". However, there has been minimal research on gastronomy tourism, specifically in Pakistan. The primary purpose of this study is to measure the level of satisfaction in a tourist destination and furthermore consider gastronomy as a component of visitor motivation. A survey of 307 tourists who had recently visited Pakistan's northern areas was undertaken to conduct the study. This has enabled us to better understand the variables that influence the behaviors and attitudes of tourists toward this popular tourist attraction. Gastronomy motivation impacts tourism location selection, and gastronomic experience influences satisfaction, according to the research. Specifically, tourists show a keen interest in gastronomic experiences after feeling satisfied with the destination and local foods, developing loyalty toward the destination.

Artificial Intelligence-Based Solution in Personalized Computer-Aided Arthroscopy of Shoulder Prostheses.

BACKGROUND: Early recognition of prostheses before reoperation can reduce perioperative morbidity and mortality. Because of the intricacy of the shoulder biomechanics, accurate classification of implant models before surgery is fundamental for planning the correct medical procedure and setting apparatus for personalized medicine. Expert surgeons usually use X-ray images of prostheses to set the patient-specific apparatus. However, this subjective method is time-consuming and prone to errors. METHOD: As an alternative, artificial intelligence has played a vital role in orthopedic surgery and clinical decision-making for accurate prosthesis placement. In this study, three different deep learning-based frameworks are proposed to identify different types of shoulder implants in X-ray scans. We mainly propose an efficient ensemble network called the Inception Mobile Fully-Connected Convolutional Network (IMFC-Net), which is comprised of our two designed convolutional neural networks and a classifier. To evaluate the performance of the IMFC-Net and state-of-the-art models, experiments were performed with a public data set of 597 de-identified patients (597 shoulder implants). Moreover, to demonstrate the generalizability of IMFC-Net, experiments were performed with two augmentation techniques and without augmentation, in which our model ranked first, with a considerable difference from the comparison models. A gradient-weighted class activation map technique was also used to find distinct implant characteristics needed for IMFC-Net classification decisions. RESULTS: The results confirmed that the proposed IMFC-Net model yielded an average accuracy of 89.09%, a precision rate of 89.54%, a recall rate of 86.57%, and an F1.score of 87.94%, which were higher than those of the comparison models. CONCLUSION: The proposed model is efficient and can minimize the revision complexities of implants.

Physicians' Perception About the Side Effects of COVID-19 Vaccine and Their Role in Vaccination Program.

INTRODUCTION: The Coronavirus disease 2019 (COVID-19) caused many problems in the health sector. Effective and safe vaccines are the only tool to combat the COVID-19 disease. Researchers and regulatory agencies have made efforts to develop such vaccines and healthcare professionals were prioritized for the vaccination program as first-line care providers. Considering this prioritization, we aimed to assess the physicians' perceptions regarding the side effects of the COVID-19 vaccine. METHODS: An interview-based study was conducted from May 5 May to November 5, 2021 (6 months) in a teaching hospital in the capital city of Islamabad, Pakistan. RESULTS: Among the 200 physicians who agreed to participate in the interview, 74% were male. A total of 94% did not experience any side effects after receiving the COVID-19 vaccine, regardless of the type of vaccine; 6% experienced side effects. Furthermore, 90% of physicians were afraid of side effects due to the high number of vaccine doses. CONCLUSION: Conclusively, physicians had a positive perception regarding the COVID-19 vaccine. Healthcare authorities and related departments must take precautions to ensure the effective and safe COVID-19 vaccine to change the perceptions of physicians who had a negative perception. This will not only reduce the reluctance among physicians regarding administering COVID-19 vaccine, but will also boost and facilitate the vaccination program for the general public as well.

HIF-1 stabilization in T cells hampers the control of Mycobacterium tuberculosis infection.

The hypoxia-inducible factors (HIFs) regulate the main transcriptional pathway of response to hypoxia in T cells and are negatively regulated by von Hippel-Lindau factor (VHL). But the role of HIFs in the regulation of CD4 T cell responses during infection with M. tuberculosis isn't well understood. Here we show that mice lacking VHL in T cells (Vhl cKO) are highly susceptible to infection with M. tuberculosis, which is associated with a low accumulation of mycobacteria-specific T cells in the lungs that display reduced proliferation, altered differentiation and enhanced expression of inhibitory receptors. In contrast, HIF-1 deficiency in T cells is redundant for M. tuberculosis control. Vhl cKO mice also show reduced responses to vaccination. Further, VHL promotes proper MYC-activation, cell-growth responses, DNA synthesis, proliferation and survival of CD4 T cells after TCR activation. The VHL-deficient T cell responses are rescued by the loss of HIF-1α, indicating that the increased susceptibility to M. tuberculosis infection and the impaired responses of Vhl-deficient T cells are HIF-1-dependent.