Adenovirus Type 40 Host Range in Tissue Culture: Replication and Gene Expression in INT407 Cells

The enteric adenoviruses show a restricted host range in tissue culture. Adenovirus type 40 (Ad40) can not be passaged in conventional human cell lines used for other adenovirus serotypes, but will grow in cells constitutively expressing Ad2 or Ad5 ElB 55K function (KB 16 & 293 cells respectively). Preliminary experiments showed that Ad40 could also be propagated in INT407 cells which were not known to express El proteins. The aim of the work described here was to characterise the growth of Ad40 in INT407 cells by examining virus replication and gene expression compared to other permissive (KB16) and semi-permissive cell lines (HeLa & A549). A timecourse of DNA replication confirmed that INT407 cells were permissive for Ad40. The pattern of DNA replication was similar on KB 16 and INT407 cells although the yield from KB16 cells was slightly higher than INT407 cells; the lowest yield was from HeLa. In addition INT407 cells could partially complement the growth of an Ad2/5 55K mutant virus. These results suggest that ElB 55K may be complemented or that the requirement for 55K may vary in INT407 cells. The kinetics of Ad40 growth were studied by fluorescent focus assay using a monoclonal antibody against Ad5 hexon. Ad40 exhibited one hit kinetics on permissive cells (INT407 & KB 16) and two hit kinetics on semi-permissive cells (A549 & HeLa). There was a marked difference in the number of productively infected cells between cell types with fewer cells productively infected at 48h p.i. on semi-permissive cell lines. The entry events of the Ad40 replicative cycle (i.e. attachment and internalisation) were investigated to determine if any defects at this stage might contribute to the Ad40 growth phenotype on the different cell lines. A similar number of Ad40 particles attached and were internalised on INT407 and HeLa cells. These results could not explain why INT407 cells were more permissive for Ad40 growth than HeLa cells. In a parallel experiment the attachment and internalisation of dl309 was found to be comparable to Ad40 on these cell lines. In order to explain the Ad40 growth phenotype in the different cell types ElB gene expression was analysed by Northern blotting and SI nuclease digestion on the different cell lines. ElB 22S mRNA was observed at early times before the onset of DNA replication in INT407 and KB 16 cells, with the 14S mRNA predominantly late in infection in these cell types. In HeLa cells ElB mRNA was detected coincident with the onset of DNA replication at much reduced levels. A similar pattern to HeLa was seen on A549 cells. The pattern of ElB transcription on INT407 cells was comparable to that reported previously on KB 16 cells which is similar to Ad 12 with 14S, 22S and 9S mRNA. The lower levels of ElB mRNA and later detection in HeLa cells may indicate that the transcription from the ElB promoter is inefficient or absent early in infection in these cells. Even in permissive cells Ad40 ElB mRNA transcription is poor compared to Ad5 which may indicate inefficient promoter usage. Therefore the Ad40 ElB transcriptional regulatory region in particular the Spl binding site was studied by gel retardation assay. The ability of Ad40 and Ad2 GC box sequences (Spl binding sites) to bind either partially purified Spl or proteins from INT407 and HeLa extracts were compared. The Ad40 Spl binding site bound the Spl transcription factor but with lower affinity than the Ad2

Similarities and differences between the E5 oncoproteins of bovine papillomaviruses type 1 and type 4: Cytoskeleton, motility and invasiveness in E5-transformed bovine and mouse cells

Bovine papillomaviruses (BPVs) are oncogenic viruses. In cattle, BPV-1/2 is associated with urinary bladder cancer and BPV-4 with upper GI tract cancer. BPV E5 is a small hydrophobic protein localised in the endoplasmic reticulum (ER) and Golgi apparatus (GA). E5 is the major transforming protein of BPVs, capable of inducing cell transformation in cultured mouse fibroblasts and, in cooperation with E7, in primary bovine cells. E5-induced cell transformation is accompanied by activation of several cellular protein kinases, including growth factor receptors, and alkalinisation of endosomes and GA. We have reported that BPV E5 causes swelling and fragmentation of the GA and extensive vacuolisation of the cytoplasm. We now show that E5 from both BPV-1 and BPV-4 disturbs the actin cytoskeleton and focal adhesions in transformed bovine cells, where these morphological and behavioural characteristics are accompanied by hyperphosphorylation of the cellular phosphotyrosine kinase c-src. Both BPV-1 and BPV-4 E5 increase the motility of transformed mouse cells, but only BPV-1 E5 causes transformed mouse cells to penetrate a matrigel matrix. BPV-1 transformed mouse cells, but not BPV-4 transformed mouse cells, have hyperhpsphorylated c-src

The Development of Employees’ Knowledge Sharing Behavior through Team Innovation Climate and Organization Culture

The purpose of this research is to provide empirical evidence concerning the impact of team innovation climate on knowledge sharing behavior and individuals’ altruistic intentions in software sector. A survey base study was conducted. Survey based-data was collected from 319 employees at software center in Pakistan. The results of this study revealed that all the constructs of team innovation climate had positive impact on altruistic intention and knowledge sharing behavior. Moreover, altruistic intention and organization culture had positive impact on knowledge sharing behavior. However, it was found that one construct of team innovation climate (Participative safety) did not have impact on organization culture. The present research contributes to the literature pertaining to the psychosocial sides of knowledge sharing behaviors. Keywords: Team innovation climate, altruistic intention, organization culture, knowledge sharing behavior

Mechanism of Corrosion and Erosion Resistance of Plasma‐ Sprayed Nanostructured Coatings

There has been a dramatic increase in recent years in a demand for tough, wear‐resistant, abrasion, erosion, and corrosion‐resistant coatings for petroleum, chemical, aerospace industry, and processes encountering harsh environments such as paper and pulp equipment (the ball valve for high‐pressure leaching). Whereas sufficient information on mechanical properties, such as abrasion, wear, and fatigue, has been gathered over the years, work on the resistance of these coatings to erosion and corrosion is seriously lacking. In the work reported, it has been shown that nanostructured TiO2 coatings offer superior physical and mechanical properties compared to conventional TiO2 coatings. Three different types of plasma‐sprayed titanium dioxide coated samples on mild steel substrate were employed for investigation. The feedstocks used were Sulzer Metco nanopowders designated as AE 9340, AE 9342, and AE 9309. Powder 9340 was a precursor. The corrosion resistance of nanostructured TiO2 coating was dictated largely by surface structure and morphology. The distribution and geometry of splat lamellae, contents of unmelted nanoparticles, and magnitude of porosity are the important factors that affect corrosion resistance. TiO2 showed excellent resistance to corrosion in 3% NaCl. The maximum corrosion rate was observed to be 4 mils per year as shown by polarization potential and weight loss studies. The erosion‐corrosion resistance of the plasma‐sprayed nanostructured titanium dioxide coatings depends largely upon the characteristics of feed powder and its reconstitution. Dense, uniform, and evenly dispersed nanostructured constituents provide a high coating integrity, which offers high resistance to erosion‐corrosion. A mechanism of erosion‐corrosion is explained in the chapter with a schematic diagram. The findings show that the nanostructured TiO2 coatings offer superior resistance to corrosion, erosion, and environmental degradation

Current statins show calcium channel blocking activity through voltage gated channels

BACKGROUND: Statins are used for treatment of hypercholestremia. Common adverse reports associated with use of statins are generalized bodyache, rhabdomyolysis, muscles weakness and gastrointestinal disorders. The current work is an attempt to explain how smooth muscles of gastrointestinal tissues are affected by the current statins (Simvastatin, atorvastatin, fluvastatin and rosuvastatin). METHODS: Effects of the current statins were studied on spontaneous activity of isolated rabbits’ jejunal preparations. Different molar concentrations (10(−12)–10(−2)M) of the statins were applied on spontaneously contracting rabbits’ jejunal preparations. As statins relaxed spontaneous activity, so we tested the statins on KCl (80 mM) induced contractions in similar test concentrations. Positive relaxant statins were tested again through construction of Calcium Concentration Response Curves (CCRCs) in the absence and presence of the statins using verapamil, a standard calcium channel blocker. CCRCs of statins were compared with CCRCs of verapamil. RESULTS: Simvastatin, atorvastatin, fluvastatin and rosuvastatin relaxed the spontaneous and KCl-induced contractions. IC(50) for simvastatin on spontaneous rabbit’s jejunal preparations is −5.08 ± 0.1 Log 10 M. Similarly, IC(50) for KCl-induced contractions is −4.25 ± 0.01 Log 10 M. Mean IC(50) (Log 10 M) for atorvastatin on spontaneous rabbit’s jejunal preparations and KCl-induced contractions are −5.19 ± 0.07 and −4.37 ± 0.09, respectively. Fluvastatin relaxed spontaneous activity of rabbits’ jejunal preparations with an IC(50) (Log 10 M) −4.5 ± 0.03. Rosuvastatin relaxed spontaneous as well as KCl (80 mM) induced contractions with respective IC(50) (Log 10 M) −3.62 ± 0.04 and −4.57 ± 0.06. In case of CCRCs, tissues pre-treated with 4.6 μg/ml of simvastatin, have IC(50) = −1.84 ± 0.03 [log (Ca(++)) M] vs control IC(50) = −2.54 ± 0.04 [log (Ca(++)) M]. Similarly, atorvastatin, fluvastatin and rosuvastatin produced significant right shift in IC(50) for CCRCs (P ≤ 0.05). In case of verapamil, IC(50) for control curves is −2.45 ± 0.06 [log (Ca (++)) M], while IC(50) in presence of verapamil (0.1 μM) is −1.69 ± 0.05 [log (Ca (++)) M]. Statins produced right shift in the IC(50) of CCRCs(.) The effects of statins are like that of effects of verapamil, a standard calcium channel blocker. CONCLUSIONS: Our findings suggest that current statins have calcium antagonistic effects that act on voltage gated calcium channels that may provide a rationale for cause muscle weakness and gastrointestinal disorders

Spatiotemporal distribution patterns of climbers along an abiotic gradient in Jhelum District, Punjab, Pakistan

Climber–abiotic parameter interactions can have important ramifications for ecosystem’s functions and community dynamics, but the extent to which these abiotic factors influence the spatial distributions of climber communities in the western Himalayas is unknown. The purpose of this study was to examine the taxonomic diversity, richness, and distribution patterns of climbers in relation to abiotic variables in the Jhelum District. The data were collected from 120 random transects between 2019 and 2021, from 360 sites within triplet quadrats (1080 quadrats), and classification and ordination analyses were used to categorize the sample transects. A total of 38 climber species belonging to 25 genera and 11 families were recorded from the study area. The Convolvulaceae were the dominant family (26.32%), followed by the Apocynaceae (21.05%), and Leguminosae (15.79%). The majority of the climbers were herbaceous in nature (71.05%), followed by woody (23.68%). Based on the relative density, the most dominant species was Vicia sativa (12.74). The majority of the species flowered during the months of March–April (28.04%), followed by August–September (26.31%). Abiotic factors have a significant influence on the distribution pattern and structure of climbers in the study area. The results show that the climbers react to the biotic environment in different ways. The findings will serve as the foundation for future botanical inventories and will be crucial for understanding the biological, ecological, and economic value of climbers in forest ecosystems. This will help forest management, conservation, and ecological restoration in the Himalayas

Global, regional, and national incidence of six major immune-mediated inflammatory diseases: findings from the global burden of disease study 2019

BACKGROUND: The causes for immune-mediated inflammatory diseases (IMIDs) are diverse and the incidence trends of IMIDs from specific causes are rarely studied. The study aims to investigate the pattern and trend of IMIDs from 1990 to 2019. METHODS: We collected detailed information on six major causes of IMIDs, including asthma, inflammatory bowel disease, multiple sclerosis, rheumatoid arthritis, psoriasis, and atopic dermatitis, between 1990 and 2019, derived from the Global Burden of Disease study in 2019. The average annual percent change (AAPC) in number of incidents and age standardized incidence rate (ASR) on IMIDs, by sex, age, region, and causes, were calculated to quantify the temporal trends. FINDINGS: In 2019, rheumatoid arthritis, atopic dermatitis, asthma, multiple sclerosis, psoriasis, inflammatory bowel disease accounted 1.59%, 36.17%, 54.71%, 0.09%, 6.84%, 0.60% of overall new IMIDs cases, respectively. The ASR of IMIDs showed substantial regional and global variation with the highest in High SDI region, High-income North America, and United States of America. Throughout human lifespan, the age distribution of incident cases from six IMIDs was quite different. Globally, incident cases of IMIDs increased with an AAPC of 0.68 and the ASR decreased with an AAPC of −0.34 from 1990 to 2019. The incident cases increased across six IMIDs, the ASR of rheumatoid arthritis increased (0.21, 95% CI 0.18, 0.25), while the ASR of asthma (AAPC = −0.41), inflammatory bowel disease (AAPC = −0.72), multiple sclerosis (AAPC = −0.26), psoriasis (AAPC = −0.77), and atopic dermatitis (AAPC = −0.15) decreased. The ASR of overall and six individual IMID increased with SDI at regional and global level. Countries with higher ASR in 1990 experienced a more rapid decrease in ASR. INTERPRETATION: The incidence patterns of IMIDs varied considerably across the world. Innovative prevention and integrative management strategy are urgently needed to mitigate the increasing ASR of rheumatoid arthritis and upsurging new cases of other five IMIDs, respectively. FUNDING: The Global Burden of Disease Study is funded by the Bill and Melinda Gates Foundation. The project funded by Scientific Research Fund of Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital (2022QN38)

Global burden of chronic respiratory diseases and risk factors, 1990–2019: an update from the Global Burden of Disease Study 2019

Background: Updated data on chronic respiratory diseases (CRDs) are vital in their prevention, control, and treatment in the path to achieving the third UN Sustainable Development Goals (SDGs), a one-third reduction in premature mortality from non-communicable diseases by 2030. We provided global, regional, and national estimates of the burden of CRDs and their attributable risks from 1990 to 2019. Methods: Using data from the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2019, we estimated mortality, years lived with disability, years of life lost, disability-adjusted life years (DALYs), prevalence, and incidence of CRDs, i.e. chronic obstructive pulmonary disease (COPD), asthma, pneumoconiosis, interstitial lung disease and pulmonary sarcoidosis, and other CRDs, from 1990 to 2019 by sex, age, region, and Socio-demographic Index (SDI) in 204 countries and territories. Deaths and DALYs from CRDs attributable to each risk factor were estimated according to relative risks, risk exposure, and the theoretical minimum risk exposure level input. Findings: In 2019, CRDs were the third leading cause of death responsible for 4.0 million deaths (95% uncertainty interval 3.6–4.3) with a prevalence of 454.6 million cases (417.4–499.1) globally. While the total deaths and prevalence of CRDs have increased by 28.5% and 39.8%, the age-standardised rates have dropped by 41.7% and 16.9% from 1990 to 2019, respectively. COPD, with 212.3 million (200.4–225.1) prevalent cases, was the primary cause of deaths from CRDs, accounting for 3.3 million (2.9–3.6) deaths. With 262.4 million (224.1–309.5) prevalent cases, asthma had the highest prevalence among CRDs. The age-standardised rates of all burden measures of COPD, asthma, and pneumoconiosis have reduced globally from 1990 to 2019. Nevertheless, the age-standardised rates of incidence and prevalence of interstitial lung disease and pulmonary sarcoidosis have increased throughout this period. Low- and low-middle SDI countries had the highest age-standardised death and DALYs rates while the high SDI quintile had the highest prevalence rate of CRDs. The highest deaths and DALYs from CRDs were attributed to smoking globally, followed by air pollution and occupational risks. Non-optimal temperature and high body-mass index were additional risk factors for COPD and asthma, respectively. Interpretation: Albeit the age-standardised prevalence, death, and DALYs rates of CRDs have decreased, they still cause a substantial burden and deaths worldwide. The high death and DALYs rates in low and low-middle SDI countries highlights the urgent need for improved preventive, diagnostic, and therapeutic measures. Global strategies for tobacco control, enhancing air quality, reducing occupational hazards, and fostering clean cooking fuels are crucial steps in reducing the burden of CRDs, especially in low- and lower-middle income countries

Global estimates on the number of people blind or visually impaired by age-related macular degeneration: a meta-analysis from 2000 to 2020

Background: We aimed to update estimates of global vision loss due to age-related macular degeneration (AMD). Methods: We did a systematic review and meta-analysis of population-based surveys of eye diseases from January, 1980, to October, 2018. We fitted hierarchical models to estimate the prevalence of moderate and severe vision impairment (MSVI; presenting visual acuity from <6/18 to 3/60) and blindness ( < 3/60) caused by AMD, stratified by age, region, and year. Results: In 2020, 1.85 million (95%UI: 1.35 to 2.43 million) people were estimated to be blind due to AMD, and another 6.23 million (95%UI: 5.04 to 7.58) with MSVI globally. High-income countries had the highest number of individuals with AMD-related blindness (0.60 million people; 0.46 to 0.77). The crude prevalence of AMD-related blindness in 2020 (among those aged ≥ 50 years) was 0.10% (0.07 to 0.12) globally, and the region with the highest prevalence of AMD-related blindness was North Africa/Middle East (0.22%; 0.16 to 0.30). Age-standardized prevalence (using the GBD 2019 data) of AMD-related MSVI in people aged ≥ 50 years in 2020 was 0.34% (0.27 to 0.41) globally, and the region with the highest prevalence of AMD-related MSVI was also North Africa/Middle East (0.55%; 0.44 to 0.68). From 2000 to 2020, the estimated crude prevalence of AMD-related blindness decreased globally by 19.29%, while the prevalence of MSVI increased by 10.08%. Conclusions: The estimated increase in the number of individuals with AMD-related blindness and MSVI globally urges the creation of novel treatment modalities and the expansion of rehabilitation services