Mohammad E. Shubair

Detection of Chlamydia trachomatis and Mycoplasma hominis, genitalium and Ureaplasma urealyticum by polymerase chain reaction in patients with sterile pyuria.

PURPOSE Chlamydia trachomatis and Mycoplasma hominis, Mycoplasma genitalium, and Ureaplasma urealyticum are associated with various diseases of the urogenital tract, but they are usually not detected by routine microbiological diagnosis. To determine the occurrence of Chlamydia trachomatis, Mycoplasma hominis, Mycoplasma genitalium, and Ureaplasma urealyticum in patients with sterile pyuria. MATERIAL/METHODS Sterile pyuria urine samples collected during the period from February 2006 to April 2007 were tested by polymerase chain reaction (PCR) for the presence of C. trachomatis, M. hominis, M. genitalium, and U. urealyticum using specific primers for each species. A total of 200 sterile pyuria samples selected from about 2400 urine samples attending the genitourinary clinic at Al-Shifa hospital, Gaza, during the period February 2006 to April 2007 and were analyzed for routine urine examination and cultured on MacConkey agar, blood agar, and sabouraud agar to detect the presence of bacteria and Candida. The 200 samples (96 male, 104 female; aged >or=18 years) containing more than 10 leukocytes / HPF and negative for culture (showing no significant growth after 24 hr) were tested by PCR for C. trachomatis and M. hominis, M. genitalium, and U. urealyticum. RESULTS C. trachomatis was detected in 20 samples (10%), U. urealyticum in 10 samples (5%), M. hominis in 6 samples (3%) and M. genitalium in 2 samples (1%). The difference in occurrence of C. trachomatis was statistically insignificant between males and females (P=0.509), but it was significant (P=0.008) for U. urealyticum. M. hominis was detected only in samples collected from female patients. On the other hand, M. genitalium was detected only in men. CONCLUSION PCR testing of sterile pyuria showed a significant number of C. trachomatis, Mycoplasma, and Ureaplasma infections. Consequently, PCR is recommended for the detection of those microorganisms in the urine samples of sterile pyuria patients.

Effects of chlorhexidine gluconate douche on normal vaginal flora.

The effects of a 0.5% aqueous chlorhexidine gluconate (CHG) douche on the normal vaginal flora of twenty healthy nonpregnant volunteers were investigated. The douche was applied in a premeasured 180-ml quantity daily for 7 consecutive days. Specimens for aerobic and anaerobic culture of the vaginal flora were obtained on 3 occasions from each volunteer, once before and twice after CHG use. The culture results were analyzed qualitatively and quantitatively. A blood sample was also collected within 24 h of the last CHG application to determine serum CHG concentration. Lactobacillus spp., Gardnerella vaginalis and Staphylococcus epidermidis were the most prevalent aerobic bacteria at all phases of the study and Bacteroides spp. were the most prevalent anaerobes. The composition of the normal flora was essentially the same 30 days after the last dose as the pretreatment flora. Small amounts of CHG were detected in the serum of all but one subject. No untoward effects on the participants were observed. Candida albicans counts were slightly higher, but prevalence was not significantly increased at the end of the study.

Prevalence of Chlamydia trachomatis among women attending gynecology and infertility clinics in Gaza, Palestine.

BACKGROUND Chlamydia trachomatis is an obligate intracellular bacterium characterized by a biphasic developmental cycle of replication. The organism is recognized as one of the major causes of sexually transmissible human bacterial infection throughout the world. Since there have been no previous studies dealing with chlamydial diagnosis in Palestine, this study was conducted to determine the prevalence of C. trachomatis infection among women attending gynecology and infertility clinics. METHODS Endocervical swabs were collected from 109 women, aged 18-52 years (median 29 years), attending gynecology and infertility clinics in Gaza. These specimens were processed using molecular (polymerase chain reaction, PCR) and enzyme immunoassay (EIA; IDEIA PCE Chlamydia) techniques. RESULTS The results obtained show that the overall prevalence rate of C. trachomatis was 20.2%. The sensitivity was 73% for the EIA, 86% for the MOMP (major outer membrane protein gene)-based PCR, and 100% for the plasmid-based PCR. Meanwhile the specificity was 94% for the EIA, 98% for the plasmid-based PCR, and 100% for the MOMP-based PCR. In multivariate analysis, only cervical discharge was significantly associated with positivity for C. trachomatis (adjusted odds ratio 5.6, 95% confidence interval 2.0-15.5; p=0.001). CONCLUSIONS The study revealed that a significant proportion of Palestinian women expressed evidence of exposure to C. trachomatis. Women with cervicitis are more likely to have been previously infected or exposed to Chlamydia infection. Furthermore, PCR proved to be superior and more efficient in the diagnosis of C. trachomatis than EIA.

Hemolysis and Mediterranean G6PD mutation (c.563 C>T) and c.1311 C>T polymorphism among Palestinians at Gaza Strip

BACKGROUND The G6PD c.563 C>T deficient mutation is endemic among Mediterranean populations but its clinical significance is not well delineated. We set up to estimate the proportion of G6PD deficient children presenting with hemolytic anemia at Al Nasser Pediatric Hospital at Gaza Strip, Palestine. We then established the prevalence of c.563T Mediterranean mutation and its linkage to c.1311 C>T polymorphism in this population. DESIGN AND METHODS G6PD deficiency was identified in children presenting with hemolytic anemia at Al Nasser Pediatric Hospital by spectrophotometric measurement of G6PD activity. G6PD exon 6 and exon 11 were amplified from genomic DNA and evaluated for c.563T mutation by sequencing and the c.1311T polymorphism by restriction fragment analysis. Seventy X-chromosomes (60 males and 5 females) from G6PD deficient patients and 40 X-chromosomes from a control group known to be not G6PD deficient were tested. RESULTS Over 80% of these children presenting with hemolytic anemia were G6PD deficient and 34% of these had the Mediterranean G6PD deficient variant. The allelic frequencies of Mediterranean c.563T and c.1311T polymorphisms among G6PD deficient patients were 0.33 and 0.38 respectively. The c.1311T polymorphism was linked in 95.2% of patients with the Mediterranean mutation, an allele frequency of 0.87, compared to the control non-G6PD deficient group with an allele frequency of 0.18. CONCLUSIONS We conclude that G6PD deficiency accounts for majority of hemolytic anemia encountered in Gaza children treated at Al Nasser Pediatric Hospital Emergency department. The Mediterranean mutation c.563T, while not accounting for a majority of G6PD deficiency, is common among G6PD deficient Gaza Strip Palestinians and is frequently, but not always, linked to the c.1311T polymorphism. This work provides a foundation for the population screening of Palestinians for G6PD deficiency and for investigations of ancestral origin of the Mediterranean variant in world populations.

Growth Inhibition of Candida albicans and Other Medically Important Yeasts by Vaginal Contraceptive Products

The antifungal effects of two commercially available spermicidal gels (Conceptrol, produced by Ortho Pharmaceutical, Raritan, N.J., and Koromex, produced by Schmid Laboratories, Little Falls, N.J.) as well as pure nonoxynol-9 and boric acid (both components of vaginal contraceptive products) were tested against 50 clinical yeast isolates by the agar dilution method. The formulated products exerted comparable dose-dependent inhibitory effects against all yeasts tested. A 3-fold dilution of the formulated spermicidal products inhibited 90% of the yeast strains tested. To determine if the antifungal effect was due to the spermicidal detergent nonoxynol-9, this compound was tested for antifungal activity but was completely ineffective against Candida albicans in concentrations up to 10%. Boric acid, present in at least one of the products (Koromex), inhibited representative yeasts at a concentration of 0.4%. The relationship of pH and oxygen tension to inhibition by the commercial spermicides was also investigated. The pH values tested ranged from 4 to 7 and had little effect on inhibition; anaerobiosis at pH 7 slightly reduced the inhibitory activity of Conceptrol gel.

Favism, the commonest form of severe hemolytic anemia in Palestinian children, varies in severity with three different variants of G6PD deficiency within the same community

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a common genetic abnormality known to predispose to acute hemolytic anemia (AHA), which can be triggered by certain drugs or infection. However, the commonest trigger is fava beans (Vicia faba) ingestion, causing AHA (favism), which may be life-threatening especially in children. G6PD deficiency is genetically highly heterogeneous, as nearly 200 different mutations have been observed. We have investigated the hematological features of acute favism in the Palestinian Gaza community that is characterized by the polymorphic coexistence of three different G6PD deficiency genes (G6PD A-, G6PD Cairo, G6PD Med). We have found by comparison to the general population (485 adults and 466 newborns) that children with favism, in terms of relative frequency, G6PD A- was under-represented, whereas G6PD Med was over-represented. We also found that the severity of anemia was significantly greater with G6PD Med and G6PD Cairo than with G6PD A-; and with G6PD Cairo, compared to the other two variants, there was greater hyperbilirubinemia, as well as persistence of mild anemia and reticulocytosis for as long as 4months after recovery from favism. This is the first report determining a differential impact of different G6PD mutations on the clinical features of favism in the same population and the same environment.

Possible association of 3′ UTR +357 A>G, IVS11-nt 93 T>C, c.1311 C>T polymorphism with G6PD deficiency

ABSTRACT Background: Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a common X-linked inherited enzymopathic disorder affecting more than 500 million people worldwide. It has so far been linked to 217 distinct genetic variants in the exons and exon–intron boundaries of the G6PD gene, giving rise to a wide range of biochemical heterogeneity and clinical manifestations. Objectives: Reports from different settings suggested the association of intronic and other mutations outside the reading frame of the G6PD gene with reduced enzyme activity and presenting clinical symptoms. The present study aimed to investigate any association of other variations apart of the exonic or exonic intronic boundaries in the development of G6PD deficiency. Methods: Sixty-seven unrelated Palestinian children admitted to the pediatric hospital with hemolytic crises due to G6PD deficiency were studied. Results: In our Palestinian cohort of 67 [59 males (M) and 8 females (F)] G6PD-deficient children, previously hospitalized for acute hemolytic anemia due to favism, molecular sequencing of the G6PD gene revealed four cases (3M and 1F) that did not have any of the variants known to cause G6PD deficiency, but the 3′ UTR c.*+357A>G (rs1050757) polymorphism in association with IVS 11 (c.1365-13T>C; rs2071429), and c.1311C>T (rs2230037). Conclusion: We now provide an additional evidence form Palestinian G6PD-deficient subjects for a possible role of 3′ UTR c.*+357 A>G, c.1365-13T>C, and/or c.1311C>T polymorphism for G6PD deficiency, suggesting that not only a single variation in the exonic or exonic intronic boundaries, but also a haplotype of G6PD should considered as a cause for G6PD deficiency.