The impact of gastrointestinal tract infection on acquiring dengue fever virus infection and haemorrhagic fever in Jeddah region, Saudi Arabia.

This study aimed to investigate the relationship between gastrointestinal tract infection and dengue hemorrhagic fever. Dengue hemorrhagic fever is a syndrome caused by the dengue virus and primarily affects children below ten years of age and is spread by the Aedes aegypti mosquito. Gastrointestinal tract infection is a bacterial and parasitic infection that leads to gastrointestinal tract inflammation which involves the small intestine and the stomach. The relationship between the two can be manifested by gastrointestinal bleeding, acute pancreatitis, and fulminant liver failure. In this research work, 600 blood and feces samples of different ages and sex (7-8 worms) were collected from Jeddah city. From the blood samples, serum was made and stored at -20°C until use. The frozen sera samples were investigated for sero-detection of DENV-NS1 antigen as a rapid, sensitive, and cost-effective test to detect asymptomatic acute DENV-infected donors and anti-DENV IgM and IgG antibodies. Feces samples were processed for the detection of parasites. The data acquired from these samples of all the 600 participants were analyzed and interpreted, followed by statistical analysis using GraphPad Prism 5.0 software. All the values were considered significant, which showed a value of less than 0.05. Results were expressed as with the range. This article documents that gastrointestinal tract manifestations frequently occur among patients with dengue hemorrhagic fever. There are close relationships between gastrointestinal tract infection and dengue hemorrhagic fever. In current work, it was established that dengue fever leads to gastrointestinal tract bleeding in the presence of intestinal parasites. Therefore, failure to identify the patients with this infection early enough can lead to an increased morbidity and mortality rate.

Defining a murine ovarian cancer model for the evaluation of conditionally-replicative adenovirus (CRAd) virotherapy agents.

BACKGROUND: Virotherapy represents a promising approach for ovarian cancer. In this regard, conditionally replicative adenovirus (CRAd) has been translated to the context of human clinical trials. Advanced design of CRAds has sought to exploit their capacity to induce anti-tumor immunization by configuring immunoregulatory molecule within the CRAd genome. Unfortunately, employed murine xenograft models do not allow full analysis of the immunologic activity linked to CRAd replication. RESULTS: We developed CRAds based on the Ad5/3-Delta24 design encoding cytokines. Whereas the encoded cytokines did not impact adversely CRAd-induced oncolysis in vitro, no gain in anti-tumor activity was noted in immune-incompetent murine models with human ovarian cancer xenografts. On this basis, we explored the potential utility of the murine syngeneic immunocompetent ID8 ovarian cancer model. Of note, the ID8 murine ovarian cancer cell lines exhibited CRAd-mediated cytolysis. The use of this model now enables the rational design of oncolytic agents to achieve anti-tumor immunotherapy. CONCLUSIONS: Limits of widely employed murine xenograft models of ovarian cancer limit their utility for design and study of armed CRAd virotherapy agents. The ID8 model exhibited CRAd-induced oncolysis. This feature predicate its potential utility for the study of CRAd-based virotherapy agents.

Studies on the protective effect of the artichoke (Cynara scolymus) leaf extract against cadmium toxicity-induced oxidative stress, hepatorenal damage, and immunosuppressive and hematological disorders in rats.

Our objective was to explore the protective effect of artichoke leaf extract (ALE) against cadmium (Cd) toxicity-induced oxidative organ damage in rats. Male albino Wistar rats were divided into four equal groups of eight animals each. The first group was assigned as a control. Groups 2-4 were orally administered with ALE (300 mg/kg bw), Cd (CdCl2, 100 mg/L drinking water), and ALE plus Cd, respectively, daily for 4 weeks. After treatment with Cd, the liver and kidney malondialdehyde (MDA) increased significantly compared with the control rats. The sera interleukin (IL)-1ß, tumor necrosis factor (TNF-α), and IL-10, liver transaminase, urea, creatinine, and peripheral neutrophil count were significantly increased in Cd-exposed rats compared to the control group. The reduced glutathione (GSH), glutathione peroxidase (GPX), superoxide dismutase (SOD), and catalase (CAT) decreased in the liver and kidney in Cd-exposed group. In combination treatment, Cd and ALE significantly improved immune response, an antioxidant system, and hepatorenal function with a significant decline in MDA. In conclusion, ALE ameliorates the immunosuppressive and hepatorenal oxidative injury stimulated by Cd in rats. These results suggest that artichoke has shown promising effects against adverse effects of Cd toxicity.

Combinatorial strategies based on CRAd-IL24 and CRAd-ING4 virotherapy with anti-angiogenesis treatment for ovarian cancer.

BACKGROUND: A major hurdle incurrent to the human clinical application of conditionally replicative adenovirus (CRAd)-based virotherapy agents is their limited therapeutic efficacy. In this study we evaluated whether arming our previously reported Ad5/3Δ24 CRAd vector containing a 24-base pair deletion in the E1A conserved region 2, which allows selective replication within Rb-p16-deficient tumor cells, to express therapeutic genes could improve oncolytic virus potency in ovarian cancer cells. We choose to assess the therapeutic benefits achieved by virus-mediated expression of interleukin 24 (IL-24), a cytokine-like protein of the IL-10 family, and the inhibitor of growth 4 (ING4) tumor suppressor protein. RESULTS: The generated CRAd-IL24 and CRAd-ING4 vectors were tested in ovarian cancer cell lines in vitro to compare their replication, yield, and cytotoxic effects with control CRAd Ad5/3∆24 lacking the therapeutic gene. These studies showed that CRAd-IL24 infection resulted in significantly increased yield of infectious particles, which translated to a marked enhancement of virus-induced cytotoxic effects as compared to CRAd-ING4 and non-armed CRAd. Testing CRAd-IL24 and CRAd-ING4 vectors combined together did not revealed synergistic effects exceeding oncolytic potency of single CRAD-IL24 vector. Both CRAds were also tested along with anti-VEGF monoclonal antibody Avastin and showed no significant augmentation of viral cytolysis by anti-angiogenesis treatment in vitro. CONCLUSIONS: Our studies validated that arming with these key immunomodulatory genes was not deleterious to virus-mediated oncolysis. These findings thus, warrant further preclinical studies of CRAd-IL24 tumoricidal efficacy in murine ovarian cancer models to establish its potential utility for the virotherapy of primary and advanced neoplastic diseases.

Combined therapy with oncolytic adenoviruses encoding TRAIL and IL-12 genes markedly suppressed human hepatocellular carcinoma both in vitro and in an orthotopic transplanted mouse model.

BACKGROUND: Gene-based virotherapy mediated by oncolytic viruses is currently experiencing a renaissance in cancer therapy. However, relatively little attention has been given to the potentiality of dual gene virotherapy strategy as a novel therapeutic approach to mediate triplex anticancer combination effects, particularly if the two suitable genes are well chosen. Both tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and interleukin-12 (IL-12) have been emerged as promising pharmacological candidates in cancer therapy; however, the combined efficacy of TRAIL and IL-12 genes for treatment of human hepatocellular carcinoma (HCC) remains to be determined. METHODS: Herein, we investigated the therapeutic efficacy of concurrent therapy with two armed oncolytic adenoviruses encoding human TRAIL gene (Ad-ΔB/TRAIL) and IL-12 gene (Ad-ΔB/IL-12), respectively, on preclinical models of human HCC, and also elucidated the possible underlying mechanisms. The effects of Ad-ΔB/TRAIL+Ad-ΔB/IL-12 combination therapy were assessed both in vitro on Hep3B and HuH7 human HCC cell lines and in vivo on HCC-orthotopic model established in the livers of athymic nude mice by intrahepatic implantation of human Hep3B cells. RESULTS: Compared to therapy with non-armed control Ad-ΔB, combined therapy with Ad-ΔB/TRAIL+Ad-ΔB/IL-12 elicited profound anti-HCC killing effects on Hep3B and HuH7 cells and on the transplanted Hep3B-orthotopic model. Efficient viral replication and TRAIL and IL-12 expression were also confirmed in HCC cells and the harvested tumor tissues treated with this combination therapy. Mechanistically, co-therapy with Ad-ΔB/TRAIL+Ad-ΔB/IL-12 exhibited an enhanced effect on apoptosis promotion, activation of caspase-3 and-8, generation of anti-tumor immune response evidenced by upregulation of interferon gamma (IFN-γ) production and infiltration of natural killer-and antigen presenting cells, and remarkable repression of intratumor vascular endothelial growth factor (VEGF) and cluster of differentiation 31 (CD31) expression and tumor microvessel density. CONCLUSIONS: Overall, our data showed a favorable therapeutic effect of Ad-ΔB/TRAIL+Ad-ΔB/IL-12 combination therapy against human HCC, and may therefore constitute a promising and effective therapeutic strategy for treating human HCC. However, further studies are warranted for its reliable clinical translation.

Maternal colonization of group B streptococcus: prevalence, associated factors and antimicrobial resistance.

BACKGROUND AND OBJECTIVES: Group B streptococcus (GBS, Streptococcus agalactiae) can be transferred during delivery to neonates from mothers who are colonized with GBS in the genital tract. GBS can cause sepsis and meningitis in newborns. This study was conducted to determine GBS colonization rates among pregnant women and the antibiotic sensitivity patterns. DESIGN AND SETTING: Prospective descriptive study at the Maternity and Children Hospital, Makkah. PATIENTS AND METHODS: Vaginal swabs from 1328 pregnant women (>=35 weeks of gestation) attending antenatal clinic were cultured in Todd-Hewitt broth supplemented with gentamicin and nalidixic acid. After 36 hours of incubation, subculture was made onto sheep blood agar and incubated in 5% carbon dioxide for 18 to 24 hours. A Microscan Walk Away system was used for the identification and antibiotic susceptibility of GBS isolates. Each isolate was also tested for group B by using latex slide agglutination test. Information such as maternal age, gestational age and parity was collected using a predesigned questionnaire. RESULTS: The study population ranged between ages 17-47 years. The GBS colonization in all age groups was found to be 13.4%. A higher colonization rate was seen in pregnant women > 40 years of age (27.4%). Women with gestational age > 42 weeks were colonized (25%) more frequently that women with a gestational age from 41-42 weeks (20.2%). An increased rate of colonization was found in women who delivered > 5 times and no colonization in women who delivered once. All GBS isolates were 100% sensitive to penicillin G, ampicillin and vancomycin. Erythromycin and clindamycin showed resistance-15.7% and 5.1%, respectively. CONCLUSION: The high prevalence of GBS colonization in pregnant women demands for screening in women attending an antenatal clinic so that intrapartum antimicrobial prophylaxis can be offered to all women who are colonized with GBS, thus preventing its transfer to the newborn.

MALDI-TOF mass spectrometry and identification of new bacteria species in air samples from Makkah, Saudi Arabia.

BACKGROUND: During the Hajj season, respiratory symptoms are very common among pilgrims. Here, we investigated the viable bacterial population in air samples collected around the slaughterhouses used during the Hajj. METHODS AND RESULTS: We collected air samples on three days from four different sites: slaughterhouses at Al-Kakia, Al-Meaisim and Al-Sharaia, and from a waste disposal area designated for the remnants of slaughter. Samples were cultured on blood agar plates for 48 h, and bacterial isolates were identified using MALDI-TOF MS. A dendrogram using the spectra of the unidentified bacterial species was constructed, and PCR amplification and sequencing of the 16S rRNA gene was performed for one isolate per cluster. In total, 2500 colonies appeared on the nutrient agar plates, and 244 were purified for further analysis. Good identification was obtained for 202 (83%) isolates by MALDI-TOF MS. The most common genera were Bacillus (n=94, 45%) and Staphyloccocus (n=55, 26%). Poor identification was obtained for 42 (17%) isolates, and their spectra clustering revealed that these isolates belonged to 10 species. Four of these were considered to be new species. CONCLUSIONS: During the Hajj, the air was contaminated by many environmental bacterial agents, and MALDI-TOF MS was successfully adapted for their rapid identification.

Genetic diversity of the pandemic influenza A (H1N1) virus in Saudi Arabia.

INTRODUCTION: Pandemic influenza A (H1N1) virus emerged and spread globally in the spring of 2009. Saudi Arabia also witnessed a severe H1N1 pandemic virus epidemic with considerable morbidity and mortality in different parts of the kingdom beginning in June 2009. The influenza A(H1N1)pdm09 virus was detected in samples collected between May 2009 and November 2010 from Makkah region. This study provides data on the viral diagnosis and genetic diversity of hemagglutinin (HA) and neuraminidase (NA) genes of influenza A (H1N1)pdm09 virus from Saudi Arabia. METHODOLOGY: Nasopharyngeal swabs from 100 clinically infected patients in the peak of the outbreak were collected from Makkah region and processed for viral diagnosis by viral culture and real-time polymerase chain reaction (PCR). HA and NA genes of 10 selected samples were sequenced and analyzed. RESULTS: A total of 100 samples were collected; only 10 samples were found to be positive for influenza A virus infection by real-time PCR. Nucleotide sequence analysis of the HA and NA genes of influenza A (H1N1) from Saudi Arabia showed significant similarities with selected isolates. The phylogenetic tree constructed for both HA and NA genes formed close clusters with selected reference isolates. CONCLUSIONS: Nucleotide sequence analysis and phylogenetic relationships of the HA and NA genes of influenza A (H1N1) virus from Saudi Arabia with selected reference isolates indicates that they were genetically close and most probably originated from influenza A(H1N1)pdm09.

Paper money and coins as potential vectors of transmissible disease.

Paper currency and coins may be a public health risk when associated with the simultaneous handling of food and could lead to the spread of nosocomial infections. Banknotes recovered from hospitals may be highly contaminated by Staphylococcus aureus. Salmonella species, Escherichia coli and S. aureus are commonly isolated from banknotes from food outlets. Laboratory simulations revealed that methicillin-resistant S. aureus can easily survive on coins, whereas E. coli, Salmonella species and viruses, including human influenza virus, Norovirus, Rhinovirus, hepatitis A virus, and Rotavirus, can be transmitted through hand contact. Large-scale, 16S rRNA, metagenomic studies and culturomics have the capacity to dramatically expand the known diversity of bacteria and viruses on money and fomites. This review summarizes the latest research on the potential of paper currency and coins to serve as sources of pathogenic agents.

Clinically significant Coagulase Negative Staphylococci and their antibiotic resistance pattern in a tertiary care hospital.

OBJECTIVES: To identify various species of coagulase negative staphylococci involved in neonatal septicaemia and determine their antimicrobial resistance pattern. METHODS: The prospective descriptive study was carried out from January 2012 to October 2013, at Umm Al-Qura University, Makkah, Saudi Arabia, and comprised clinical isolates of coagulase negative staphylococci recovered from the blood of neonates at Maternity and Children Hospital, Makkah..The identification of species and antibiotic sensitivity for each isolate was done using Microscan Walk Away system. RESULTS: Of the 190 clinical isolates S. epidermidis was the most common species found 144(75.8%).The overall drug resistance among the species ranged from 1.6% to 99.5% to all the drugs tested, except to vancomycin and linezolid which were 100% sensitive.The highest drug resistance was exhibited by penicillin 189 (99.5%), ampicillin 188 (99%), oxacillin 178 (93.6%) and augmentin 177 (93%). The minimum drug resistance was shown by synercid 4 (2.2%) and daptomycin 3 (1.6%). All species were 100% resistant to penicillin and ampicillin, except S. hyicus and one isolate of S. hominis-homin which was sensitive to ampicillin. CONCLUSION: High rates of antibiotic resistance was seen among coagulase negative staphylococci to commonly-used antibiotics and majority of them were methicillin-resistant. The newer drugs tested can be used as an alternative.

Human immunodeficiency virus type-1 (HIV-1) disease progression and viral activity: a seroepidemiological and molecular study.

OBJECTIVE: To determine the frequency and epidemiological characterization of human immunodeficiency virus type-1 (HIV-1) infection, HIV disease progression, immune status and viral activity. STUDY DESIGN: Descriptive study. PLACE AND DURATION OF STUDY: Department of Microbiology, University of the Punjab and Institute of Public Health, Lahore, from September 2005 to August 2008. METHODOLOGY: The study enrolled samples from general population, high risk groups and spouses of HIV+ deport workers with criteria; positive double enzyme linked immunosorbent assay (ELISA) and positive western blot. Immune status and viral activity was determined by cluster determinants (CD4+ and CD8+) cell count, ratio of CD4+/CD8+ on flow cytometer, and HIV RNA viral load on polymerase chain reaction (PCR). RESULTS: A total of 116 HIV+ untreated subjects enrolled after screening of 2260 blood samples. The seroprevalence rate in general population, high risk individuals and spouses of HIV+ deport workers was found 0%, 0.4% and 26% respectively. The CD4+ cell count was found 533/mm3 (range 12-1800/mm3) and plasma viral load 27,122 copies/ml (range 00-40,621). The CD4+/CD8+ ratios < 0.5, < 1, < 1.5 and < 2 appeared as 17.2%, 30.2%, 51.7% and 0.9% respectively. Significant correlation was observed between plasma viral load, CD4+ count and CD4+/CD8+ ratio (p = 0.001). CD4+ T-cell counts < 200 cells/mm3 was found in 23 HIV+ patients. CONCLUSION: There was a low frequency of HIV in the general population and high risks groups as compared to very high frequency in spouses of HIV+ deport workers with significant correlation of viral activity and immune status.

Application of ventilator care bundle and its impact on ventilator associated pneumonia incidence rate in the adult intensive care unit.

OBJECTIVE: To reduce ventilator associated pneumonia (VAP) incidence rate, lessen the cost of care, and correlate VAP bundles compliance with VAP incidence rate. METHODS: This study was a prospective longitudinal study conducted on adult intensive care unit (ICU) patients at Hera General Hospital, Makkah, Kingdom of Saudi Arabia from January to December 2010. The following Institute for Healthcare Improvement VAP prevention bundle was applied: head-of-bed elevation; daily sedation-vacation along with a readiness-to-wean assessment; peptic ulcer disease (PUD) prophylaxis; and deep venous thrombosis (DVT) prophylaxis. RESULTS: Among a total of 2747 patients, the bundle compliance rate in January 2010 was 30%, and reached to 100% in December 2010, while the overall rate was 78.9%. The individual bundle compliance rates were as follows: head-of-bed elevation - 99.9%; daily sedation vacation - 88.9%; PUD prophylaxis - 94.9%; and DVT prophylaxis - 85.7%. At the beginning, VAP rate was 2.5/1000 ventilator days, and reduced to 0.54 in the next month. The overall VAP incidence rate in 2010 was found to be 1.98 with a reduction of 1.41 by comparing with the same data of year 2009 collected retrospectively. The total reduction cost in one year was $154,930. A significant correlation was found between the VAP rate and its bundle compliance (p=0.001). Most frequent pathogens found were Pseudomonas aeruginosa (30.8% of all isolates) followed by Acinetobacter baumannii (27.7%), and methicillin-resistant Staphylococcus aureus (15.4%). CONCLUSION: Application of VAP prevention bundle reduced the VAP incidence rate and lowered the cost of care.

Profile of bacterial pneumonia during Hajj.

BACKGROUND & OBJECTIVES: The congregation of a large number of people during Hajj seasons from different parts of the world in overcrowded conditions within a confined area for a long period of time presents many public health challenges and health risks. One of the main health problems of the crowding is ease transmission of pneumonia by air droplets. This study was aimed to determine the most common causes of bacterial pneumonia during the 2005 Hajj season and to relate the findings with clinical conditions. METHODS: A total of 141 patients with suspected pneumonia from the three main tertiary care hospitals in Makkah, Saudi Arabia, were investigated during Hajj season, 2005. Sputum and serum samples were collected and investigated for the possible presence of typical or atypical causative agents. RESULTS: Of the 141 clinically suspected pneumonia cases, 76 (53.9%) were confirmed positive by microbiological tests. More than 94 per cent of the confirmed cases were in the age group >50 yr, and 56.6 per cent of the cases were men. The most frequent isolates were Candida albicans (28.7%) and Pseudomonas aeruginosa (21.8%), followed by Legionella pneumophila (14.9%) and Klabsiella pneumoniae (9.2%). More than one causative pathogens were isolated in 15 patients (16.3%), and 55 per cent of patients were diabetic. INTERPRETATION & CONCLUSIONS: Clinicians should be aware that typical pneumonia treatment regimens may not work well during the Hajj season due to the wide variety of isolated organisms. This necessitates taking a sputum sample before starting treatment for identification and sensitivity testing. Special precautions need to be taken for >50 yr old patients.

Multi-drug resistant Ewingella americana.

We report a case of pneumonia due to multi-drug resistant Ewingella americana in a young patient admitted in the Intensive Care Unit of Hera General Hospital, Makkah, Saudi Arabia with severe head injury in a road traffic accident. He was an Indonesian pilgrim who had traveled to the Kingdom of Saudi Arabia to perform Hajj in December 2007. Ewingella americana was identified to be the pathogen of pneumonia with clinical signs and symptoms along with positive radiological findings.

Clinical profile and outcome of hospitalized patients during first outbreak of dengue in Makkah, Saudi Arabia.

OBJECTIVE: To describe clinical profile of patients with dengue virus infection hospitalized at a single center during the first outbreak of dengue in Makkah, Saudi Arabia from April to July 2004. METHODS: Clinical information and laboratory abnormalities of patients with suspected dengue infection were collected by a standardized data collection sheet and review of medical records. Dengue virus infection was confirmed by a positive IgM capture ELISA or RT-PCR. RESULTS: Of the 160 clinically suspected patients, 91 were confirmed (64 by IgM ELISA, 14 by RT-PCR and 13 by both) to have dengue virus infection. Dengue serotypes 2 and 3 were identified in 19 and 4 patients respectively. Most patients were young adults with median age of 26 (range=6-94) years and male:female ratio of 1.5:1. The common symptoms were fever (100%), malaise (83%), musculoskeletal pain (81%), headache (75%), nausea (69%), vomiting (65%) and abdominal pain (48%). According to World Health Organization (WHO) classification (10 patients were excluded due to lack of serial hematocrits), 75 (93%) had dengue fever (DF) and 6 (7%) had dengue hemorrhagic fever (DHF). Only one patient with DHF was in pediatric age group. Twenty-one patients (5 with DHF and 16 with DF) developed one or more clinical complications that included bleeding (14), shock (4), seizures (3), acute renal failure (2), meningo-encephalitis (1), and secondary bacterial infection (1). Only one patient with shock had dengue shock syndrome (DSS) by WHO classification. Development of clinical complications was significantly associated with absence of musculoskeletal pain (p-value=0.03), lower platelet counts (p-value=0.03) and higher serum aspartate aminotransferase levels (p-value=0.04). The median duration of symptoms and hospitalization was 8 days (range=3-18) and 4 days (range=1-10) respectively. No mortality was noted. CONCLUSION: Occurrence of dengue virus infection in Makkah, Saudi Arabia is documented. Continued surveillance and effective vector control programs are warranted due to unique population dynamics of Makkah that receives millions of pilgrims annually from all over the world.