Carbon monoxide-releasing molecule-3 (CORM-3; Ru(CO)3Cl(glycinate)) as a tool to study the concerted effects of carbon monoxide and nitric oxide on bacterial flavohemoglobin Hmp: applications and pitfalls.

CO and NO are small toxic gaseous molecules that play pivotal roles in biology as gasotransmitters. During bacterial infection, NO, produced by the host via the inducible NO synthase, exerts critical antibacterial effects while CO, generated by heme oxygenases, enhances phagocytosis of macrophages. In Escherichia coli, other bacteria and fungi, the flavohemoglobin Hmp is the most important detoxification mechanism converting NO and O2 to the ion nitrate (NO3(-)). The protoheme of Hmp binds not only O2 and NO, but also CO so that this ligand is expected to be an inhibitor of NO detoxification in vivo and in vitro. CORM-3 (Ru(CO)(3)Cl(glycinate)) is a metal carbonyl compound extensively used and recently shown to have potent antibacterial properties. In this study, attenuation of the NO resistance of E. coli by CORM-3 is demonstrated in vivo. However, polarographic measurements showed that CO gas, but not CORM-3, produced inhibition of the NO detoxification activity of Hmp in vitro. Nevertheless, CO release from CORM-3 in the presence of soluble cellular compounds is demonstrated by formation of carboxy-Hmp. We show that the inability of CORM-3 to inhibit the activity of purified Hmp is due to slow release of CO in protein solutions alone i.e. when sodium dithionite, widely used in previous studies of CO release from CORM-3, is excluded. Finally, we measure intracellular CO released from CORM-3 by following the formation of carboxy-Hmp in respiring cells. CORM-3 is a tool to explore the concerted effects of CO and NO in vivo.

Leverage of Salvadora persica and Pulicaria undulata extracts in Escherichia coli-challenged broiler chickens.

Escherichia coli (E. coli) is a significant challenge in the poultry industry due to their related use of antimicrobial compounds and the drastic losses in production and livability. This study investigated the preventive impacts of dietary supplementation of Salvadora persica (SP) and/or Pulicaria undulata (PU) extracts on growth traits, biochemical and immune parameters, and related gene expression of E. coli-infected broilers. A total of 120 one-day-old Cobb broilers were used. The chicks were allocated into eight equal groups (3 replicates/ group; 5 chicks per each replicate) as follows: G1; control negative, G2; SP-treated, G3; PU-treated, G4; SP/PU-treated, G5; E. coli infected, G6; E. coli infected and SP-treated, G7; E. coli infected and PU-treated, G8; E. coli infected and SP/PU-treated groups. Results revealed significant improvement in average body weight (ABW), average weight gain (AWG) and feed conversion ratio (FCR) in broilers fed diets supplemented with SP and/or PU compared to control and E. coli infected groups. Moreover, significant (P < 0.05) reduction in ALT, AST, creatinine, and uric acid was reported in other treated groups compared to the single E. coli-infected broilers. On the contrary, a significant increase (P < 0.05) in serum immunoglobulin and protein concentration was also reported in treated groups when compared to E. coli-infected untreated group. In addition, feeding broilers with SP and/or PU significantly improved (P < 0.05) the relative weight of immune-related organs and gene expression of TLR-15, with subsequent down-regulation of IL-1ß and TNF-α mRNA transcripts. Supplementing broilers with dietary SP and/or PU could be promising in the prevention of E. coli infection via stimulating significant improvement of immune-related gene expression, immune-related organ weight, and down-regulation of inflammatory-related genes, with subsequent enhancement of the growth performance of broiler chickens.

Evaluation of the Antimicrobial Activity of ZnO Nanoparticles against Enterotoxigenic Staphylococcus aureus.

Staphylococcus aureus (S. aureus) is a Gram-positive bacteria considered one of the leading causes of community and hospital-acquired illnesses or public health concerns. Antibiotic resistance in this microorganism is one of the greatest issues in global health care. The use of metal nanoparticles and their oxides is one of the potential approaches to combating bacteria resistance to antibiotics. The antibacterial properties of ZnO NPs against enterotoxigenic S. aureus were studied. ZnO NPs were tested in vitro by agar diffusion test. They resulted in 26 and 22 mm zones of inhibition for a size of 20 nm and a concentration of 20 mM against 105 and 107 CFU/mL S. aureus, respectively. The MIC of ZnO NPs of various sizes, 20 and 50 nm, with 105 CFU/mL was 2.5 and 5 mM, respectively. MIC with 107 CFU/mL was five mM for 20 and 50 nm ZnO NPs. Further, the highest growth reduction percentage, 98.99% in the counts of S. aureus was achieved by ZnO NPs of size 20 nm and concentration of 10 mM. Moreover, the obtained ELISA results indicated a significantly decreased concentration of enterotoxin A with all concentrations and sizes of ZnO NPs. PCR analysis showed a significant effect on sea gene in response to ZnO NPs treatments leading to loss of the gene, unlike the unaffected nuc gene. Moreover, morphological changes and cell shape distortion were detected by scanning electron microscope for bacterial cells treated with ZnO NPs.

Levels and predictors of fear and health anxiety during the current outbreak of COVID-19 in immunocompromised and chronic disease patients in Saudi Arabia: A cross-sectional correlational study.

Coronavirus disease 2019 (COVID-19) infection has become a clinical threat to healthy people as well as immunocompromised patients and those with pre-existing chronic diseases around the world. This study, which used a cross-sectional correlational design, aimed to assess the levels of fear and health anxiety and to investigate their predictors during the current outbreak of COVID-19 in immunocompromised and chronic disease patients in Saudi Arabia. Sociodemographic and clinical data, fear of COVID-19, and health anxiety measurements were collected by online surveys from June 15 to July 15, 2020. Univariate and multiple linear regression analysis was used to identify predictors. A total of 1,030 patients in 13 provinces in Saudi Arabia completed the questionnaire. A significant number of patients with chronic diseases experienced considerable levels of fear and anxiety during the COVID-19 outbreak. It was found that 21.44% of participants met the criteria for anxiety cases, and 19.4% were considered borderline anxiety cases. In regression analysis, significant predictors of fear and health anxiety were female gender, lower education, middle-aged, divorced or widowed, receiving immunosuppressants, type of chronic disease (Crohn's disease, hypertension, and cardiovascular diseases), and media use as a source of knowledge about COVID-19. Immunocompromised and chronic disease patients are vulnerable to fear and anxiety during epidemic infectious diseases such as COVID-19. Optimizing this population's compliance with appropriate infection prevention and control strategies is crucial during the infectious outbreaks to ensure their safety, to decrease the risk of infection and serious complications, and reduce their fear and health anxiety. Effective positive psychological interventions and support strategies also need to be immediately implemented to increase psychological resilience and improve the mental health of these patients. Due to the COVID-19 outbreak, chronic disease patients in Saudi Arabia need special attention from health authorities, policymakers, and healthcare professionals to manage maladaptive forms of health anxiety and fear.