Upregulation of interleukin-19 in severe asthma: a potential saliva biomarker for asthma severity

Interleukin (IL)-19, a designated IL-20 subfamily cytokine, has been implicated in inflammatory disorders including rheumatoid arthritis, psoriasis and, lately, asthma. Here, through the analysis of transcriptomic datasets of lung tissue of large asthma cohorts, we report that IL-19 expression is upregulated in asthma and correlates with disease severity. The gene expression of IL-19 was significantly higher in lung tissue from patients with severe and mild/moderate asthma compared to healthy controls. IL-19 protein level, however, was significantly higher in the blood and saliva of patients with severe asthma compared to mild/ moderate subgroups as measured by ELISA assay. IL-19 protein level was not affected by corticosteroid treatment in plasma. Our data provide insights into the potential use of IL-19 as a saliva marker for asthma severity and a potential therapeutic target

Towards an antibody mediated resistance against Geminiviruses

In this study, single chains (scFvs) against key proteins of the Tomato leaf curl virus-India were generated using phage display and/or hybridoma technologies. Purified AV1 (coat protein) and AC1 (replicase enhancer) viral proteins were used for mice immunization and screening of phage display libraries and hybridoma clones. The binding properties of the isolated scFvs were determined by western blot, ELISA and BIACORE. The missing nucleotide found in the hybridoma derived scFv-SAV against AV1 was restored and the resulting scFv (scFv-RW-AV) showed better binding properties than scFv-SAV. Four scFvs against AV1 were derived from the naïve Tomlinson I library. Two of these scFvs carried a stop codon. The stop codon was mutated in vitro to either code for glutamine (Q) or glutamic acid (E). The Q-mutated scFvs showed higher expression level but less binding activity than the E-mutated scFvs. Additionally, two scFvs against AC1 were selected. All scFvs were cloned and expressed in tobacco leaves as N-terminal fusion with the GFP for cytosolic expression in tobacco leaves. Additionally, two scFv-GFP fusions were targeted to the nucleus by a nuclear localization signal (NLS) introduced at the N-terminus of the scFvs. Both scFv-GFP fusion proteins were exclusively localized in the nucleus. The AV1 proteinwas also expressed in tobacco leaves as N-terminal fusion with DsRed. The AV1-DsRed fusion was completely localized in the nucleus. The binding activity of the AV1-specific scFvs was tested by the co-expression of the scFv- GFP fusion and AV1-DsRed. Only the hybridoma derived scFv showed clear binding and the protein complex was targeted to the nucleus by the AV1 NLS. The data presented in this study demonstrate the successful expression of specific and functional scFvs against the viral proteins in the reducing environment of the plant cytosol. Such a scFv is a potential candidate to interfere with the viral life cycle and thus abolishing or delaying the viral infection

Towards an antibody mediated resistance against Geminiviruses

In this study, single chains (scFvs) against key proteins of the Tomato leaf curl virus-India were generated using phage display and/or hybridoma technologies. Purified AV1 (coat protein) and AC1 (replicase enhancer) viral proteins were used for mice immunization and screening of phage display libraries and hybridoma clones. The binding properties of the isolated scFvs were determined by western blot, ELISA and BIACORE. The missing nucleotide found in the hybridoma derived scFv-SAV against AV1 was restored and the resulting scFv (scFv-RW-AV) showed better binding properties than scFv-SAV. Four scFvs against AV1 were derived from the naïve Tomlinson I library. Two of these scFvs carried a stop codon. The stop codon was mutated in vitro to either code for glutamine (Q) or glutamic acid (E). The Q-mutated scFvs showed higher expression level but less binding activity than the E-mutated scFvs. Additionally, two scFvs against AC1 were selected. All scFvs were cloned and expressed in tobacco leaves as N-terminal fusion with the GFP for cytosolic expression in tobacco leaves. Additionally, two scFv-GFP fusions were targeted to the nucleus by a nuclear localization signal (NLS) introduced at the N-terminus of the scFvs. Both scFv-GFP fusion proteins were exclusively localized in the nucleus. The AV1 proteinwas also expressed in tobacco leaves as N-terminal fusion with DsRed. The AV1-DsRed fusion was completely localized in the nucleus. The binding activity of the AV1-specific scFvs was tested by the co-expression of the scFv- GFP fusion and AV1-DsRed. Only the hybridoma derived scFv showed clear binding and the protein complex was targeted to the nucleus by the AV1 NLS. The data presented in this study demonstrate the successful expression of specific and functional scFvs against the viral proteins in the reducing environment of the plant cytosol. Such a scFv is a potential candidate to interfere with the viral life cycle and thus abolishing or delaying the viral infection

Selection criteria for high-yielding and early-flowering bread wheat hybrids under heat stress.

Hybrid performance during wheat breeding can be improved by analyzing genetic distance (GD) among wheat genotypes and determining its correlation with heterosis. This study evaluated the GD between 16 wheat genotypes by using 60 simple sequence repeat (SSR) markers to classify them according to their relationships and select those with greater genetic diversity, evaluate the correlation of the SSR marker distance with heterotic performance and specific combining ability (SCA) for heat stress tolerance, and identify traits that most influence grain yield (GY). Eight parental genotypes with greater genetic diversity and their 28 F1 hybrids generated using diallel crossing were evaluated for 12 measured traits in two seasons. The GD varied from 0.235 to 0.911 across the 16 genotypes. Cluster analysis based on the GD estimated using SSRs classified the genotypes into three major groups and six sub-groups, almost consistent with the results of principal coordinate analysis. The combined data indicated that five hybrids showed 20% greater yield than mid-parent or better-parent. Two hybrids (P2 × P4) and (P2 × P5), which showed the highest performance of days to heading (DH), grain filling duration (GFD), and GY, and had large genetic diversity among themselves (0.883 and 0.911, respectively), were deemed as promising heat-tolerant hybrids. They showed the best mid-parent heterosis and better-parent heterosis (BPH) for DH (-11.57 and -7.65%; -13.39 and -8.36%, respectively), GFD (12.74 and 12.17%; 12.09 and 10.59%, respectively), and GY (36.04 and 20.04%; 44.06 and 37.73%, respectively). Correlation between GD and each of BPH and SCA effects based on SSR markers was significantly positive for GFD, hundred kernel weight, number of kernels per spike, harvest index, GY, and grain filling rate and was significantly negative for DH. These correlations indicate that the performance of wheat hybrids with high GY and earliness could be predicted by determining the GD of the parents by using SSR markers. Multivariate analysis (stepwise regression and path coefficient) suggested that GFD, hundred kernel weight, days to maturity, and number of kernels per spike had the highest influence on GY

Human Adenovirus Molecular Characterization in Various Water Environments and Seasonal Impacts in Riyadh, Saudi Arabia

The regular monitoring of water environments is essential for preventing waterborne virus-mediated contamination and mitigating health concerns. We aimed to detect human adenovirus (HAdV) in the Wadi Hanifah (WH) and Wadi Namar (WN) lakes, King Saud University wastewater treatment plant (KSU-WWTP), Manfouha-WWTP, irrigation water (IW), and AnNazim landfill (ANLF) in Riyadh, Saudi Arabia. HAdV hexon sequences were analyzed against 71 HAdV prototypes and investigated for seasonal influence. ANLF had the highest HAdV prevalence (83.3%). Remarkably, the F species of HAdV, especially serotype 41, predominated. Daily temperature ranges (22–45 °C and 10–33 °C) influenced the significance of the differences between the locations. The most significant relationship of ANLF and IW to WH and KSU-WWTP was found at the high-temperature range (p = 0.001). Meanwhile, WN was most correlated to ANLF at the low-temperature range (p < 0.0001). Seasonal influences on HAdV prevalence were insignificant despite HAdV’s high prevalence in autumn and winter months, favoring low temperatures (high: 22–25 °C, low: 14–17 °C) at five out of six locations. Our study provides insightful information on HAdV prevalence and the circulating strains that can address the knowledge gap in the environmental impacts of viruses and help control viral diseases in public health management

Ectopic Expression of a Wheat R2R3-Type MYB Gene in Transgenic Tobacco Enhances Osmotic Stress Tolerance via Maintaining ROS Balance and Improving Root System Architecture

Water scarcity is a critical cause of plant yield loss and decreased quality. Manipulation of root system architecture to minimize the impact of water scarcity stresses may greatly contribute towards an improved distribution of roots in the soil and enhanced water and nutrient uptake abilities. In this study, we explored the potential of TaMYB20 gene, a wheat gene belonging to the R2R3-MYB transcription factor family, to improve root system architecture in transgenic tobacco plants. The full-length TaMYB20 gene was isolated from Triticum aestivum.cv. Sakha94 and used to produce genetically engineered tobacco plants. The transgenic plants exhibited enhanced tolerance to extended osmotic stress and were able to maintain their root system architecture traits, including total root length (TRL), lateral root number (LRN), root surface area (RSa), and root volume (RV), while the wild-type plants failed to maintain the same traits. The transgenic lines presented greater relative water content in their roots associated with decreased ion leakage. The oxidative stress resulted in the loss of mitochondrial membrane integrity in the wild-type (WT) plants due to the overproduction of reactive oxygen species (ROS) in the root cells, while the transgenic lines were able to scavenge the excess ROS under stressful conditions through the activation of the redox system. Finally, we found that the steady-state levels of three PIN gene transcripts were greater in the TaMYB20-transgenic lines compared to the wild-type tobacco. Taken together, these findings confirm that TaMYB20 is a potentially useful gene candidate for engineering drought tolerance in cultivated plants

ASSESSMENT OF KNOWLEDGE, ATTITUDE, AND PRACTICE TOWARDS OSTEOPOROSIS SCREENING AMONG PRIMARY HEALTH CARE PHYSICIANS IN PRIMARY HEALTH CARE CENTERS IN MAKKAH AL-MOKARRAMAH CITY, 2023. (ACROSS SECTIONAL STUDY)

<p><strong>Background:</strong>Osteoporosis, a silent disease characterized by decreased bone density and increased fracture risk, poses a significant public health challenge. This study aims to investigate the knowledge, attitudes, and practices of Primary Health Care (PHC) physicians in MakkahAl-Mokarramah regarding osteoporosis screening. Osteoporosis is a multifactorial skeletal disorder with increasing prevalence, especially in regions with an aging population, such as Saudi Arabia. Understanding the current state of awareness and practices among PHC physicians is essential for early detection and intervention.</p><p><strong>Method</strong>:This study employs a cross-sectional design and targets PHC physicians in Makkah Al-Mokarramah in 2023. The sample size was calculated, and participants were selected through simple random sampling. Data was collected using a structured electronic questionnaire with sections on demographics, knowledge, attitudes, and practices related to osteoporosis screening.</p><p><strong>Results:</strong> The study focused on PHC physicians, with a significant proportion below the age of 30 and a university-level education. The gender distribution was nearly even, with a focus on both males and females. Regarding knowledge, the majority of participants exhibited adequate knowledge (58%) of osteoporosis, while 19.2% had poor knowledge, and 22.8% had proper knowledge. There were notable knowledge gaps, especially among those with poor knowledge. Regarding attitude,most participants believed that osteoporosis is a major health concern (85.2%) and that it is preventable (83.6%). Confidence in screening and managing osteoporosis varied based on knowledge levels, with those having proper knowledge expressing the highest confidence. Regarding practice, the study revealed variations in participants practices related to osteoporosis, including calcium and vitamin D supplementation, physiotherapy recommendations, participation in continuous medical education, and the use of tools like FRAX.</p><p><strong>Conclusion</strong>: The study reveals that while there is general awareness of osteoporosis, there are significant knowledge gaps among PHC physicians. Attitudes toward prevention are positive, but these attitudes must be translated into concrete practices, such as improved dietary habits and physical activity. We recommend further research initiatives and calls for both government and non-government organizations to intensify their efforts in promoting preventive programs, particularly tailored to the needs of the population. Overall, the study contributes to the understanding of osteoporosis awareness and behaviors, offering a foundation for public health interventions in this domain.</p><p> </p&gt

Phenotypic and genotypic analysis of pathogenic Escherichia coli virulence genes recovered from Riyadh, Saudi Arabia

The current study was carried out to evaluate the phenotypic and genotypic characterization of avian pathogenic Escherichia coli recovered from Riyadh, Saudi Arabia. During the period of 10th February–30th May 2015, 70 E. coli strains were isolated from chicken farms located in Riyadh, Saudi Arabia. All strains were tested phenotypically by standard microbiological techniques, serotyped and the virulence genes of such strains were detected by polymerase chain reaction (PCR). Most of the recovered strains from chickens belonged to serotype O111:K58 25 strains (35.7%), followed by serotype O157:H7 13 strains (18.57%), followed by serotype O114:K90 10 strains (14.29%), then serotype O126:K71 9 strains (12.9%), serotype O78:K80 8 strains (11.43%) and in lower percentage serotype O114:K90 and O119:K69 5 strains (7.14%). The virulence genotyping of E. coli isolates recovered from broilers revealed the presence of the uidA gene in all the field isolates (6 serovars) examined in an incidence of 100%, as well as the cvaC gene was also present in all field isolates (6 serovars), while the iutA gene and the iss gene were detected in 5 out of 6 field serovars in an incidence of 81.43% and 64.29%, respectively. Phenotypical examination of the other virulence factors revealed that 65 isolates were hemolytic (92.9%), as well as 15 isolates (21.42%) were positive for enterotoxin production. Meanwhile, 21 isolates (30%) were positive for verotoxin production, 58 isolates (82.86%) for the invasiveness and 31 isolates (44.29%) for Congo red binding activities of the examined serotypes. Keywords: Escherichia coli, Virulence factors, Serotyping, PCR, Avian pathogeni

Biochemical and pharmaceutical traits of Marrubium vulgare L. plants treated with plant growth-promoting bacteria and elevated CO\u2082

Abstract: The present research aimed to understand the influence of plant growth-promoting bacteria (PGPB) on various biochemical, nutritional, and pharmaceutical characteristics of Marrubium vulgare plants grown under elevated carbon dioxide (eCO(2)). To achieve this objective, a pot experiment was carried out, consisting of two treatments, namely: (i) biofertilization (Bf) by a PGPB strain (Micromonospora sp.) and (ii) two different air CO2 levels, including ambient CO2 (aCO(2)) and eCO(2) concentrations (410 and 710 mu mol CO2 mol(-1), respectively). The improvement in the photosynthesis rate of eCO(2) and Bf-treated plants can explain the increase in the production of carbohydrate. This is evidenced by a substantial rise, reaching up to + 75% and 25% in the total sugar and starch content in plants subjected to eCO(2) treatment, respectively. Additionally, eCO(2)-treated plants exhibited a remarkable 102% increase in soluble sugar synthesis, while plants subjected to Bf treatment showed a notable increase of 66%. Such modifications could be the main factor affecting plants carbon and nitrogen metabolism. Although the level of certain amino acids (such as glycine, tyrosine, and phenylalanine) in plants exhibited significant increases in response to eCO(2) and Bf, the levels of other amino acids demonstrated enhancements in plants grown under eCO(2) (e.g., histidine) or under treatments containing Bf (e.g., alanine and ornithine). Improvements in primary metabolites led to more benefits in plants treated with Bf and CO2 by boosting secondary metabolites accumulation, including phenolics (+ 27-100%), flavonoids (+ 30-92%), and essential oils (up to + 296%), as well as improved antioxidant capacity (FRAP). This remarkable effectiveness was evident in the significant increase in the biomass production, highlighting the synergistic impact of the treatments. Therefore, the interaction of Bf and eCO(2) not only induced plant biomass accumulation but also improved the nutritional and pharmaceutical value of M. vulgare plants