Human cytomegalovirus UL24 and UL43 products participate in SAMHD1 subcellular localization.

This report has analyzed the potential role of Human Cytomegalovirus (HCMV) UL24 and UL43 products in modulating the subcellular location of a host restriction factor, SAMHD1, in cells of human fibroblast origin. Recent studies have reported that the regulation of SAMHD1 is mediated by the HCMV UL97 product inside the nucleus, and by the CDK pathway when it is located in the cytoplasm of the infected cells but the viral gene products that may involve in cytosolic relocalization remain unknown yet. In the present report, we demonstrate that the HCMV UL24 product interacts with the SAMHD1 protein during infection based on mass spectrometry (MS) data and immunoprecipitation assay. The expression or depletion of the viral UL24 gene product did not affect the subcellular localization of SAMHD1 but when it coexpressed with the viral UL43 gene product, another member of the HCMV US22 family, induced the SAMHD1 cytosolic relocalization. Interestingly, the double deletion of viral UL24 and UL43 gene products impaired the cytosolic translocation and the SAMHD1 was accumulated in the nucleus of the infected cells, especially at the late stage post-infection. Our results provide evidence that the viral UL24 and UL43 gene products play a role in the SAMHD1 subcellular localization during HCMV infection. Supplementary Information: The online version contains supplementary material available at 10.1007/s13337-022-00799-3.

Human Cytomegalovirus UL24 and UL43 Cooperate to Modulate the Expression of Immunoregulatory UL16 Binding Protein 1.

The human cytomegalovirus (HCMV) UL24 and UL43 are tegument proteins that have recently been shown to interact with each other in a yeast two-hybrid system. By their overexpression in MRC5 cells, we demonstrate that these viral proteins interact with several important host proteins, especially Dicer and trans-activation response RNA binding protein. As these hots proteins are involved in regulating the production of cellular micro-RNAs, the cytomegalovirus (CMV) proteins could interfere with their actions to favor viral replication directly or through an immune escape mechanism. Double knockout of UL24 and UL43 does not show a remarkable effect on CMV entry or replication, but it significantly downregulates the expression of CMV-encoded miR-UL59, which is thought to regulate the expression of a downstream target UL16 binding protein 1 (ULBP1). Interestingly, the double knockout increases the expression of the ULBP1 recognized by the NKG2D activating receptor of natural killer cells. This study investigates the potential role of several proteins encoded by HCMV in regulating the host cellular environment to favor escape from immunity, and it also provides some basis for the future development of RNA-targeted small molecules to control HCMV infection.

Physiological regulation for enhancing biosynthesis of biofilm-inhibiting secondary metabolites in Streptomyces cellulosae.

Although the production of the secondary metabolite is frequently restricted, methods to regulate and optimize their synthesis are extremely beneficial. The current study proposes to enhance the production of antibiofilm metabolite in Streptomyces cellulosae (S. cellulosae). It was isolated from soil by growing on Gause's media and identified by colony morphology and genomic sequencing of 16S rDNA. Antibacterial and antibiofilm activities of the isolates were screened against a series of pathogenic bacteria by agar plug diffusion and 96 well microtiter plate methods, respectively. Physiological regulation of the bacterial bioactivity against biofilm formation was monitored under different cultural conditions. The isolated Streptomyces sequence analysis of the 16S rDNA was 100% identical to the sequence of S. cellulosae strain NBRC 13027. Physical (temperature and pH) and chemical (carbon, nitrogen, and minerals) culture medium factors have shown variable impacts on the growth and bioactive substances of S. cellulosae. Moreover, results of simple linear regression and correlation suggested that most of the physiological regulations with the highest response (r2= 0.85-0.99; p<0.01) and linearly (r= 0.88-0.99; p<0.01) were correlated between microbial biomass and crude extract. Lastly, under different culture growth conditions, biofilm inhibition was tested against Pseudomonas aeruginosa (P. aeruginosa). The physiological regulation results exhibited that 1 µg/mL of the extract was the most efficient concentration against biofilm formation in P. aeruginosa while 3 µg/mL is an effective bactericidal dose against P. aeruginosa. We concluded that S. cellulosae can produce antibacterial and antibiofilm metabolites. Physiological regulation is considered a powerful tool that can be used for increasing the biosynthesis of the active metabolites and biomass.

Cancer Incidence in the Kurdistan Region of Iraq: Results of a Seven-Year Cancer Registration in Erbil and Duhok Governorates.

INTRODUCTION: There are limited published data regarding the recent incidence trends of cancer in Iraqi Kurdistan. METHODS: The present study assessed the epidemiological estimates of cancer incidence, as well providing a projection of future cancer trends in the upcoming decade by analysing the population-based cancer registry between 2013 and 2019, in both the Erbil and Duhok governorates. A retrospective analysis was performed on data retrieved from the Medical Statistics Department at the Ministry of Health, Kurdistan Regional Government (KRG). RESULTS: The total number of female cancer patients was higher in both governorates, and the total incidence of patients with cancer increased by over 2x between 2013 and 2019 in Erbil and Duhok, from 73 to 174 patients/100,000 individuals for women, and 36 to 85 patients/100,000 individuals for men. Analysis indicated that the percentage of patients with cancer is projected to increase by >2x in the current decade, from 3,457 cases to 4,547 and 4,449 cases in the Erbil governorate; and from 1,365 to 2,633 and 2,737 cases in 2028 based on LSTM and bi-LTSM analysis in the Duhok governorate. Lung cancer (LC) and female breast cancer (BC) were the most prominent types of cancers diagnosed since 2013 in both the Erbil and Duhok governorates. CONCLUSION: The striking pattern of trends for both present and future cancer incidence rates require urgent solutions and comprehensive efforts to control risk factors that promote the increasing incidence of cancer in these two KRG governorates.
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Host Stress Signals Stimulate Pneumococcal Transition from Colonization to Dissemination into the Lungs.

Streptococcus pneumoniae is an asymptomatic colonizer of the nasopharynx, but it is also one of the most important bacterial pathogens of humans, causing a wide range of mild to life-threatening diseases. The basis of the pneumococcal transition from a commensal to a parasitic lifestyle is not fully understood. We hypothesize that exposure to host catecholamine stress hormones is important for this transition. In this study, we demonstrated that pneumococci preexposed to a hormone released during stress, norepinephrine (NE), have an increased capacity to translocate from the nasopharynx into the lungs compared to untreated pneumococci. Examination of NE-treated pneumococci revealed major alterations in metabolic profiles, cell associations, capsule synthesis, and cell size. By systemically mutating all 12 two-component and 1 orphan regulatory systems, we also identified a unique genetic regulatory circuit involved in pneumococcal recognition and responsiveness to human stress hormones. IMPORTANCE Microbes acquire unique lifestyles under different environmental conditions. Although this is a widespread occurrence, our knowledge of the importance of various host signals and their impact on microbial behavior is not clear despite the therapeutic value of this knowledge. We discovered that catecholamine stress hormones are the host signals that trigger the passage of Streptococcus pneumoniae from a commensal to a parasitic state. We identify that stress hormone treatment of this microbe leads to reductions in cell size and capsule synthesis and renders it more able to migrate from the nasopharynx into the lungs in a mouse model of infection. The microbe requires the TCS09 protein for the recognition and processing of stress hormone signals. Our work has particular clinical significance as catecholamines are abundant in upper respiratory fluids as well as being administered therapeutically to reduce inflammation in ventilated patients, which may explain why intubation in the critically ill is a recognized risk factor for the development of pneumococcal pneumonia.

Cardiac, Hepatic and Renal Dysfunction and IL-18 Polymorphism in Breast, Colorectal, and Prostate Cancer Patients.

INTRODUCTION: The present study aimed to determine the alterations in the serum levels of tumor markers used to evaluate cardiac, renal and liver function, and detect the interleukin (IL)-18 rs1946518 polymorphism in breast (BC), colorectal (CRC) and prostate cancer (PCa) patients. METHODS: Blood samples were collected from 65 female BC, 116 CRC, 79 PCa and 88 myocardial infarction (MI) patients, and 110 healthy individuals to determine the concentration of tumor and cardiac markers. Furthermore, the IL-18 rs1946518 polymorphism was assessed using amplification refractory mutation system (ARMS)-PCR. RESULTS: The serum levels of the tumor markers cancer antigen 15-3 (CA 15-3), carbohydrate antigen 19-9 (CA 19-9), carcinoembryonic antigen (CEA) and total prostate-specific antigen (TPSA) were significantly increased in cancer patients compared with healthy controls. Furthermore, the activity of high-sensitivity cardiac troponin T (hs-cTnT) and creatine kinase­myocardial band (CK-MB) was enhanced in MI patients, however, their activity was unchanged in cancer patients. The activity of alkaline phosphatase (ALP), and the serum concentration of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and urea were markedly elevated in CRC and PCa patients, respectively, compared with the control group. Although, no significant differences were observed in the -607 C/A polymorphism and allele frequency of IL-18 among BC, CRC patients and healthy individuals, the odds ratio (OR) was 1.75 for both C and A allele in BC patients. Therefore, the -607 C/A polymorphism could be considered as a risk factor for BC. CONCLUSION: The aforementioned results suggested that tumor markers could be considered as excellent biomarkers for the early detection of BC, CRC and PCa, whereas the concentration of liver enzymes could serve as an alternative indicator for the diagnosis of CRC and PCa. Additionally, the rs1946518 polymorphism in the IL-18 gene could be considered as a risk factor for the occurrence of BC, CRC and PCa.
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Streptococcus pneumoniae Cell Wall-Localized Trigger Factor Elicits a Protective Immune Response and Contributes to Bacterial Adhesion to the Host.

Trigger factor (TF) has a known cytoplasmic function as a chaperone. In a previous study we showed that pneumococcal TF is also cell-wall localized and this finding combined with the immunogenic characteristic of TF, has led us to determine the vaccine potential of TF and decipher its involvement in pneumococcal pathogenesis. Bioinformatic analysis revealed that TF is conserved among pneumococci and has no human homologue. Immunization of mice with recombinant (r)TF elicited a protective immune response against a pneumococcal challenge, suggesting that TF contributes to pneumococcal pathogenesis. Indeed, rTF and an anti-rTF antiserum inhibited bacterial adhesion to human lung derived epithelial cells, indicating that TF contributes to the bacterial adhesion to the host. Moreover, bacteria lacking TF demonstrated reduced adhesion, in vitro, to lung-derived epithelial cells, neural cells and glial cells. The reduced adhesion could be restored by chromosomal complementation. Furthermore, bacteria lacking TF demonstrated significantly reduced virulence in a mouse model. Taken together, the ability of rTF to elicit a protective immune response, involvement of TF in bacterial adhesion, conservation of the protein among pneumococcal strains and the lack of human homologue, all suggest that rTF can be considered as a future candidate vaccine with a much broader coverage as compared to the currently available pneumococcal vaccines.

Functional assessment of microbial superoxide dismutase isozymes suggests a differential role for each isozyme.

Microbes can have multiple enzymes that are able to catalyse the same enzymatic reactions but may differ in structure. These are known as isozymes. It is assumed that isozymes have the same functional role for cells. Contrary to this assumption, we hypothesised that isozymes can confer different functions for microbial cells despite catalysing the same reactions. To test this hypothesis, we studied the role of superoxide dismutases (SOD) in Klebsiella pneumoniae, the causative agent of several nosocomial and community-acquired infections, in infection relevant assays. SODs are responsible for detoxification of toxic superoxide radicals. K. pneumoniae genome contains three superoxide dismutase genes, sodA, sodB, and sodC coding for Mn-, Fe- and CuZn- co-factored SODs, respectively. By creating and testing single, double, and triple SOD mutants, we investigated the regulatory interactions among SOD and determined the role of each isozyme in oxidative stress resistance, biofilm formation, cell morphology, metabolism, and in vivo colonization and persistence. Our results demonstrate that SOD isozymes in K. pneumoniae have unique roles beyond oxidative stress resistance, and there is a regulatory interplay among SODs.

Flavin Reductase Contributes to Pneumococcal Virulence by Protecting from Oxidative Stress and Mediating Adhesion and Elicits Protection Against Pneumococcal Challenge.

Pneumococcal flavin reductase (FlaR) is known to be cell-wall associated and possess age dependent antigenicity in children. This study aimed at characterizing FlaR and elucidating its involvement in pneumococcal physiology and virulence. Bioinformatic analysis of FlaR sequence identified three-conserved cysteine residues, suggesting a transition metal-binding capacity. Recombinant FlaR (rFlaR) bound Fe2+ and exhibited FAD-dependent NADP-reductase activity, which increased in the presence of cysteine or excess Fe2+ and inhibited by divalent-chelating agents. flaR mutant was highly susceptible to H2O2 compared to its wild type (WT) and complemented strains, suggesting a role for FlaR in pneumococcal oxidative stress resistance. Additionally, flaR mutant demonstrated significantly decreased mice mortality following intraperitoneal infection. Interestingly, lack of FlaR did not affect the extent of phagocytosis by primary mouse peritoneal macrophages but reduced adhesion to A549 cells compared to the WT and complemented strains. Noteworthy are the findings that immunization with rFlaR elicited protection in mice against intraperitoneal lethal challenge and anti-FlaR antisera neutralized bacterial virulence. Taken together, FlaR's roles in pneumococcal physiology and virulence, combined with its lack of significant homology to human proteins, point towards rFlaR as a vaccine candidate.

Antibiotic susceptibility of vancomyin and nitrofurantoin in Staphylococcus aureus isolated from burnt patients in Sulaimaniyah, Iraqi Kurdistan.

Burns remain a significant public health problem in terms of morbidity and mortality throughout the world, especially in low and middle-income countries. Burning raptures the skin barriers that normally prevent invasion by microorganisms and infection is a major complication in burn patients. Methicillin resistant Staphylococcus aureus (MRSA) is the most important nosocomial pathogen. This retrospective analysis was conducted in the burn unit of the Department of Microbiology in the Sulamaini Plastic Surgery and Burns Hospital. The analysis is based on data collected from the medical records of 2938 burn patients, hospitalized between May 2008 and December 2011. The clinical samples were taken from various body sources for microbiological tests. Patients with a high percentage of total body surface area burnt (P<0.001) and a longer hospital stay (P<0.001) were more likely to have infection compared to other patients. In addition, among all tested antibiotics, vancomycin and nitrofurantion seem to be the most effective antibiotics for MR- SA. Furthermore there was a significant association between age and antibiotic resistance for all antibiotics except for vancomycin and nitrofurantoin. Resistance to antibiotics increased with advancing age. The wide use of antibiotics in the treatment of bacterial infections has probably led to the emergence and spread of resistant strains. Routine microbiological surveillance and careful in vitro testing prior to antibiotic use and strict adherence to hospital antibiotic policy may help in the prevention and treatment of antibiotic resistant pathogens in burn infections.