Ahmed Mohamed Ashshi

Complete genome sequencing and phylogenetic analysis of dengue type 1 virus isolated from Jeddah, Saudi Arabia

BackgroundDengue viruses (DENVs) are mosquito-borne viruses which can cause disease ranging from mild fever to severe dengue infection. These viruses are endemic in several tropical and subtropical regions. Multiple outbreaks of DENV serotypes 1, 2 and 3 (DENV-1, DENV-2 and DENV-3) have been reported from the western region in Saudi Arabia since 1994. Strains from at least two genotypes of DENV-1 (Asia and America/Africa genotypes) have been circulating in western Saudi Arabia until 2006. However, all previous studies reported from Saudi Arabia were based on partial sequencing data of the envelope (E) gene without any reports of full genome sequences for any DENV serotypes circulating in Saudi Arabia.FindingsHere, we report the isolation and the first complete genome sequence of a DENV-1 strain (DENV-1-Jeddah-1-2011) isolated from a patient from Jeddah, Saudi Arabia in 2011. Whole genome sequence alignment and phylogenetic analysis showed high similarity between DENV-1-Jeddah-1-2011 strain and D1/H/IMTSSA/98/606 isolate (Asian genotype) reported from Djibouti in 1998. Further analysis of the full envelope gene revealed a close relationship between DENV-1-Jeddah-1-2011 strain and isolates reported between 2004–2006 from Jeddah as well as recent isolates from Somalia, suggesting the widespread of the Asian genotype in this region.ConclusionsThese data suggest that strains belonging to the Asian genotype might have been introduced into Saudi Arabia long before 2004 most probably by African pilgrims and continued to circulate in western Saudi Arabia at least until 2011. Most importantly, these results indicate that pilgrims from dengue endemic regions can play an important role in the spread of new DENVs in Saudi Arabia and the rest of the world. Therefore, availability of complete genome sequences would serve as a reference for future epidemiological studies of DENV-1 viruses.

Demographic distribution and transmission potential of influenza A and 2009 pandemic influenza A H1N1 in pilgrims

INTRODUCTION The World Health Organizations persistent reporting of global outbreaks of influenza A viruses, including the 2009 pandemic swine A H1N1 strain (H1N1pdm09), justified the targeted surveillance of pilgrims during their annual congregation that pools more than two million people from around 165 nations in a confined area of Makkah city in the Kingdom of Saudi Arabia (KSA). METHODOLOGY A total of 1,600 pilgrims were included in the targeted surveillance of influenza A and the 2009 pandemic swine H1N1 strain in the Hajj (pilgrimage) season of 2010. Each pilgrim responded to a demographic and health questionnaire. Collected oropharyngeal swabs were analyzed by real-time PCR for influenza A viruses, and positive samples were further analyzed for the presence of H1N1pdm09. Fishers exact test was applied in the analysis of the significance of the distribution of influenza-positive pilgrims according to demographic characters. RESULTS A total of 120 pilgrims (7.5%) tested positive for influenza A viruses by real-time PCR. Nine out of the 120 influenza-A-positive pilgrims (7.5%) were positive for H1N1pdm09. Demographics played a significant role in those pilgrims who tested positive for influenza A. CONCLUSIONS The detection of H1N1pdm09 in pilgrims at their port of entry to the KSA was alarming, due to the high potential of trans-boundary transmission. This situation necessitates the implementation of specific prevention and control programs to limit infection by influenza A viruses.

The prevalence of dengue virus serotypes in asymptomatic blood donors reveals the emergence of serotype 4 in Saudi Arabia

BackgroundTransmission of dengue virus (DENV) through blood transfusion has been documented and hence screening for DENV during blood donation has been recently recommended by the American Association of Blood Banks and Centres of Disease Control and Prevention. DENV is endemic in the Western province of the Kingdom of Saudi Arabia (KSA) and serotypes 1, 2 and 3, but not 4, have been detected. However, little is known regarding the rates of DENV during blood donation in the kingdom. The aim of this study was therefore to measure the prevalence of dengue virus and its serotypes in eligible Saudi blood donors in the endemic Western region of KSA.MethodsThis was a cross-sectional study and serum samples were collected from 910 eligible Saudi male blood donors. DENV IgM and IgG antibodies were measured serologically by ELISA while viral serotypes were detected by a single step IVD CE certified multiplex RT-PCR kit.ResultsThe overall prevalence was 39 and 5.5% for IgG+ and IgM+, respectively. There were 12 (1.3%) with exclusively IgM+, 317 (34.8%) exclusively IgG+ and 38 (4.2%) with dual IgM+/IgG+ donors. The overall prevalence was 3.2% (n = 29) and 2.3% (n = 21) for primary and secondary infections. PCR was positive in 5.5% (n = 50) and, DENV-2 (n = 24; 48%) was the most frequent serotype and was significantly higher than DENV-1 (20%; P = 0.02) and DENV-3 (2%; P = 0.1 × 10−5) but not DENV-4 (30%; P = 0.2). There was no significant difference between both DENV-4 and DENV-1 (P = 0.4). The combination of the PCR and serology findings showed that 22 (2.4%) and 28 (3.1%) donors had primary and secondary viremic infections, respectively.ConclusionsThe detected rates of DENV by PCR suggest a potential high risk of viral transmission by blood transfusion. To the best of our knowledge, this study is the first to report the detection of DENV-4 serotype in Saudi Arabia. More studies are required to measure the precise prevalence of DENV serotypes and their potential transmission rate during blood donation in the kingdom.

Serum Activins and Follistatin during the Treatment of Chronic Hepatitis C Genotypes 1 and 4 and Their Correlations with Viral Load and Liver Enzymes: A Preliminary Report

Aims. To measure the effect of pegylated interferon-α therapy on serum activin-A, activin-B, and follistatin and their correlation with viral load and liver fibrosis in chronic hepatitis C (CHC). Methods. This study was cross-sectional and sera were collected from 165 participants classified into 7 groups: 40 healthy negative control, 33 treatment naïve patients as positive control, 19 patients at week 4, 22 at week 12, and 19 at week 24 of treatment initiation and 21 responders and 11 nonresponders at the end of 48-week treatment protocol. Serum candidate proteins were measured using ELISA and liver fibrosis was assessed by AST platelet ratio index (APRI). Results. CHC significantly increased activins and decreased follistatin compared to negative control (P < 0.05). Activin-A and follistatin levels returned to the levels of negative control group at weeks 4, 12, and 24 following treatment initiation and were significantly different from positive control (P < 0.05). Both proteins were significantly different between responders and nonresponders. Activin-A correlated positively and significantly with the viral load and APRI. Conclusion. CHC modulates serum activin-A and follistatin and they appear to be influenced by pegylated interferon-α therapy. Further studies are needed to explore the role of activins in CHC.

The prevalence of Chlamydia trachomatis and Mycoplasma genitalium tubal infections and their effects on the expression of IL-6 and leukaemia inhibitory factor in Fallopian tubes with and without an ectopic pregnancy.

This was a prospective case–control study that measured the prevalence of Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG) and Mycoplasma genitalium (MG) by an IVD CE multiplex PCR kit in fresh Fallopian tubes (FT) obtained from 96 ectopic pregnancies (EP) and 61 controls in the midluteal phase of the cycle. We later measured the expression profile of IL-6, leukaemia inhibitory factor (LIF) and their signalling molecules, in respect to the type and number of infections, by immunohistochemistry, ELISA and quantitative RT-PCR. The frequencies of CT, and MG mono- and co-infections were significantly higher in EP. IL-6, LIF, their receptors and intracellular mediators were significantly up-regulated at the gene and protein levels in positive compared with negative FTs within each group (P < 0.05). EP tubal samples with co-infections showed the highest significant expression of the candidate cytokines by all techniques (P < 0.05). CT and MG are frequent in EP and up-regulate the tubal expression of IL-6, LIF and their signalling molecules. Both cytokines could be involved in the tubal immune response against bacterial infections, as well as the pathogenesis of EP. Further studies are needed to explore the roles of IL-6 family in infection-induced tubal inflammation and EP.

Activins and Follistatin in Chronic Hepatitis C and Its Treatment with Pegylated-Interferon-α Based Therapy

Pegylated-interferon-α based therapy for the treatment of chronic hepatitis C (CHC) is considered suboptimal as not all patients respond to the treatment and it is associated with several side effects that could lead to dose reduction and/or termination of therapy. The currently used markers to monitor the response to treatment are based on viral kinetics and their performance in the prediction of treatment outcome is moderate and does not combine accuracy and their values have several limitations. Hence, the development of new sensitive and specific predictor markers could provide a useful tool for the clinicians and healthcare providers, especially in the new era of interferon-free therapy, for the classification of patients according to their response to the standard therapy and only subscribing the novel directly acting antiviral drugs to those who are anticipated not to respond to the conventional therapy and/or have absolute contraindications for its use. The importance of activins and follistatin in the regulation of immune system, liver biology, and pathology has recently emerged. This review appraises the up-to-date knowledge regarding the role of activins and follistatin in liver biology and immune system and their role in the pathophysiology of CHC.

Islamic Wet Cupping and Risk Factors of Cardiovascular Diseases: Effects on Blood Pressure, Metabolic Profile and Serum Electrolytes in Healthy Young Adult Men

Background: Wet cupping (Hejamah) has been used as alternative treatment for several diseases. Objectives: Materials and methods: 16 participants were treated with hejamah for 2 consecutive months. Blood pressure was measure before and 30 minutes after the treatment. Blood samples were collected from all participants before and 48 hours after hejamah and all participants were fasting for 12 hours before sample collection. Results: Fasting blood glucose was significantly decreased before the treatment in the second month only. The levels of serum triglycerides significantly decreased after the first treatment and remained low in the 2nd month. There was no significant difference between the different time points in total cholesterol except for the 48 hours of the second month compared to the samples collected before the procedure of the same month. There was a significant decrease in LDL and significant increase in HDL following hejamah therapy (P<0.05). Significant decrease in sodium and significant increase in potassium 48 hours following treatment with hejamah was observed in the 2 months (P<0.05). Conclusions: The performance of hejamah during fasting state could represent a useful complementary method for the regulation of diastolic blood pressure and prevention/treatment of risk factors associated with cardiovascular diseases. Further studies are required to explore the role of hejamah in controlling blood pressure and prevention of cardiovascular diseases.

Characterisation of vitamin D-related molecules and calcium-sensing receptor in human Fallopian tube during the menstrual cycle and in ectopic pregnancy

This is a prospective observational study that measures the expression of vitamin D (VD) metabolising and signalling molecules and Ca2+ sensing receptor (CaSR) in human Fallopian tube (FT) during the menstrual cycle and ectopic pregnancy (EP). Fresh FTs were obtained during total abdominal hysterectomy at the follicular (n = 16) and midluteal (n = 16) phases. Specimens from remote and implantation sites as well as trophoblastic tissues were also freshly collected from each FT with EP (n = 10). All women had normal serum VD and ionised Ca2+. The expression of VD synthesising (CYP27B1) and catalysing (CYP24A1) enzymes, binding protein (VDBP), receptor (VDR), retinoid X receptor (RXR) and CaSR was measured by immunohistochemistry and quantitative RT-PCR. All molecules, except VDBP, were significantly increased (P < 0.05) in midluteal compared with follicular samples. Remote EP sites showed significantly (P < 0.05) lower expression of CYP27B1, CYP24A1, VDR and RXR and a higher expression of VDBP and CaSR (P < 0.05) compared with midluteal samples. Significant differences were observed by immunohistochemistry between implantation and remote sites from EP for all molecules, which were also localised in the trophoblastic tissues. In conclusion, VD and calcium are under cycle-dependent regulations within human FT and they appear to play a role in tubal biology through paracrine/autocrine mode of signalling. Furthermore, EP was associated with alterations in the expression of all the studied molecules by the tubal epithelium. Further studies are needed to explore the roles of VD in tubal biology and pathogenesis of EP.

Seroprevalence of Chlamydia trachomatis, cytomegalovirus, herpes simplex virus 1 and 2 in Saudi women with normal and abnormal early pregnancy: A case control study

2 HIV patients were included in this study, who were either on maintenance hemodialysis (n=14) or were renal transplant recipients (n=10). Out of the 24, most were males (85.5%). Their spouses (n=20) were also included in this study. All subjects were interrogated via a questionnaire. All 24 patients were HIV positive. Out of the 20 spouses, 8 were found to be HIV positive (40%). Probable mode of spread of infection was enquired from the patients, 20 patients out of 24 had multiple sexual contacts (83.3%), while blood transfusion and multiple needle pricks due to drug abuse were found in one each (8.4%). 2 patients were not sure of any cause (8.4%). Out of the 20 spouses, 18 were sexually active, out of which 8 were found to be HIV positive and all were of the opinion that they had received the infection due to sexual contact with their spouses who were HIV positive CKD patients. Out of the 18 sexually active couples 12 were using condoms, while 6 spouses who were not using condoms, were found to be HIV positive. Out of the 8 HIV positive spouses, 2 had received the infection in spite of using condoms, thus implying that barrier contraception, though extremely effective, is not a full proof method of protection from the spread of HIV infection. Furthermore 15 patients were chronic alcoholics (62.5%). Most of them thought that social workers are the best way of preventing the disease (n=40, 90.5%), followed by increasing awareness through newspapers and television (n=36, 81.8%). Out of 44 subjects 30 (68.2%) thought that teaching in the school on the subject would be helpful in preventing it, while 28 (63.6%) were of the opinion that the family physicians could prevent the spread of the infection by counseling the families of HIV infected patients. Thus this study concludes that multiple sexual contacts, chronic alcohol abuse, intravenous drug abuse and spouses of patients are at high risk of contracting HIV infection and that social workers may be extremely beneficial in increasing awareness of HIV and restricting its spread.Background: In the setting of progressive immunosuppression, as with a HIV infection, the incidence of cervical dysplasia is increased. Paradoxically, among certain ethnicities and economic levels, HIV-infected women have a lower rate of screening for cervical cancer via Papanicolaou smear (Pap smear) compared to HIV-negative women. Understanding barriers to cervical cancer screening among HIV-infected women is important to develop an intervention to increase adherence to guidelines.H entry is mediated by the interaction of the trimeric envelope glycoprotein (Env) on the virus membrane surface with host cell receptors. However, Env is the only virus-specific protein on the virion surface and is essential for cell receptor interactions and subsequent virus-cell fusion. Therefore, HIV-1 Env is an important target to directly inhibit and thus block the initial steps leading to host cell infection. Our lab has synthesized peptide triazoles, a class of novel entry inhibitors. These peptides contain a substituted triazole derivative formed from a synthetically introduced azido-proline amino acid and bind to gp120 with close to nanomolar affinity. Site-directed mutagenesis and molecular dynamics simulation have shown that peptide triazole binding overlaps the CD4 binding pocket. Peptide triazoles cause cell-independent gp120 shedding, and variants containing C-terminal cysteines cause cell-free virolysis as evidenced by internal p24 capsid release. We are investigating the mode of action by which the sulfhydryl group causes irreversible inactivation. We hypothesize that the thiol interferes with conserved disulfides clustered proximal to the CD4 binding site in gp120 through “disulfide exchange”, which could deform the Env protein spike, and subsequently the viral membrane, leading to p24 release. The process of disulfide exchange has been found to be necessary for HIV viral infection.T study offers a conceptual model which certainly contributes to the literature of discrimination toward people living with HIV (PLHIV) and can provide the theoretical basis for future experimental research as much as intervention actions in order to achieve the aim of Zero discrimination. The health care practitioners’ (HCP) attitudinal factors toward PLHIV as much as the institutional factors should be undertaken in HIV discrimination researches. While the most of researches have relatively done in the field of training and wide-awakening of HCP about HIV transmission in Iran and Asian countries, the process of forming the discriminatory intention among the HCP have not been studied yet. In order to explain the factors affecting the HCPs’ discriminatory intention toward PLHIV, not only the author developed and hypothesized a model using the theory of planned behavior’s (TPB) framework but also proposed a new method of attitude measurement aims to shed a new light on predicting HIV discrimination in the health settings. The hypothesis is including: 1. The belief in the just world (BJW) among the HCPs’ affects their perception of stigma as much as their prejudice and also both of them affect the discriminatory attitude toward PLHIV 2. The perception of stigma affects the HCPs’ socio/cultural risk perception 3. The socio/cultural risk perception affects the HCPs’ discriminatory attitude 4. The transmission risk perception affects the HCPs’ discriminatory attitude 5. The discriminatory attitude affects the HCPs’ discriminatory intention toward PLHIV 6. Fuzzy method compared with the existing conventional measurement methods can measure more precisely the HCPs’ discriminatory attitudeT study determined whether socio-demographic characteristics and social support are associated with quality of life in individuals diagnosed with HIV/AIDS in Ghana. This study was guided by concepts from the intersection domains of social capital, social network, and social support theories. A convenience sample of 300 HIV/AIDS support group members were obtained via cross-sectional design survey. Data were collected from 300 participants selected because they attend support groups meetings, are a convenient sample, and also have experience in participating in research studies. The Medical Outcome Studies (MOS) HIV Health Survey and the MOS Social Support Survey (MOS-SSS) and demographic questionnaire instruments were used to assess quality of life, social support, and demographic information respectively. Multiple regression analysis was performed to determine if socio-demographic factors and social support contribute to quality of life. There was a positive association between overall social support and overall quality of life (r=0.51). The combination of socio-demographic factors and social support related to quality of life. Implications of the findings for practice, research, and policy in Ghana were discussed.T precise molecular mechanisms on how HIV-1 co-infection accelerates HCV-mediated liver disease progression are currently unknown. Our data showed that infectious HIV-1 virus particles failed to enter into human hepatocytic cell lines, and discernible virus replication was not observed, even when the hepatocytes transfected with HIV-1 proviral DNA were co-cultured with Jurkat T cells, suggesting that liver deterioration in the co-infected patients is not due to the replication of HIV-1 in the hepatocytes. Instead, HIV-1 Nef protein can be transferred from Nef-expressing or HIV-1-infected cells to hepatocytes through conduits, not exosomes, and the transferred Nef in the target hepatocytes altered lipid droplet formation, up-regulated subgenomic replicon expression of HCV, augmented reactive oxygen species (ROS) production, and enhanced ethanol-mediated up-regulation of HCV replication. Taken together, these data indicate that HIV-1 Nef plays an integral role in expedition of liver pathogenesis in the HIV-1/HCV co-infected hosts.S the beginning of the 21st century, we are facing the convergence of several epidemics. These include tobacco smoking, tuberculosis (TB) and HIV infection. These epidemics interact by way of increasing disease susceptibility and worsening outcomes. To control these interacting epidemics, it is crucial to better understand each infection and how it influences the others. The association between tobacco smoke and TB was suggested many years ago. Evidence of the impact of tobacco smoking on TB infection has been confounded by its almost universal association with poverty, overcrowding and alcohol usage. Similar pathological mechanisms induced by malnutrition, alcohol abuse and smoking may indeed all predispose an individual to TB. Although both tobacco smoking and HIV infection may be associated through their common associations with poverty and high-risk behavior, tobacco smoking appears to be an independent and important risk factor for contracting HIV. Smoking further raises the extremely high risk of contracting TB in HIV+ individuals. Individuals with HIV/AIDS are at risk for many oral health problems, particularly those who are smokers. People living with HIV/AIDS (PLWHA) are more likely to experience chronic illnesses such as cardiovascular disease and diabetes that are linked to poor oral health status. This presentation will describe the impact of HIV, TB and tobacco use on the oral health of PLWHA; it will discuss the importance of multidisciplinary health teams in the oral care for PLWHA; and it will suggest concrete steps that clinicians can take to promote positive behavioral health changes in PLWHA.

Pegylated Interferon-α Modulates Liver Concentrations of Activin-A and Its Related Proteins in Normal Wistar Rat.

Aims. To measure the expression of activin βA-subunit, activin IIA and IIB receptors, Smad4, Smad7, and follistatin in the liver and the liver and serum concentrations of mature activin-A and follistatin in normal rat following treatment with pegylated interferon-α (Peg-INF-α) and ribavirin (RBV). Materials and Methods. 40 male Wistar rats were divided equally into 4 groups: “control,” “Peg-only” receiving 4 injections of Peg-INF-α (6 µg/rat/week), “RBV-only” receiving ribavirin (4 mg/rat/day) orally, and “Peg & RBV” group receiving both drugs. The expression of candidate molecules in liver was measured by immunohistochemistry and quantitative PCR. The concentrations of mature proteins in serum and liver homogenate samples were measured using ELISA. Results. Peg-INF-α  ± RBV altered the expression of all candidate molecules in the liver at the gene and protein levels (P < 0.05) and decreased activin-A and increased follistatin in serum and liver homogenates compared with the other groups (P < 0.05). There were also significant correlations between serum and liver activin-A and follistatin. Conclusion. Peg-INF-α modulates the hepatic production of activin-A and follistatin, which can be detected in serum. Further studies are needed to explore the role of Peg-INF-α on the production of activins and follistatin by the liver and immune cells.