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.

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.

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.

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.

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.

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.