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.

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.

Molecular heterogeneity of glucose-6-phosphate dehydrogenase deficiency in Gaza Strip Palestinians.

BACKGROUND: Glucose-6-phosphate dehydrogenase (G6PD) deficiency, affecting more than 500 million people worldwide, is one of the most common of inherited disorders. There are 186 G6PD mutations published, with mutational clustering within defined ethnic/racial groups. However comprehensive molecular characterization of ethnically associated G6PD mutants and their clinical implications are lacking. DESIGN AND METHODS: Eighty unrelated Palestinian children hospitalized for hemolysis were studied. G6PD activity was determined by quantitative spectrophotometry and G6PD mutations were analyzed by sequencing of gDNA. RESULTS: 65 of 80 children (81%) had G6PD deficiency, accounting for most of the hemolytic disease in this age group. G6PD Mediterranean(c.563T), African G6PD A-(c.202A/c.376G), and G6PD Cairo(c.404C) were common with relative allele frequencies of 0.33 [1], 0.26, and 0.18 respectively. Two other variants were discovered, G6PD Beverly Hills(c.1160A) mutation, and a novel G6PD missense mutation c.536G>A (Ser179Asn), designated G6PD "Gaza". Three samples exhibited enzyme deficiency without detectable exonic or exon/intron boundary mutations. CONCLUSION: G6PD deficiency accounts for the majority of diagnoses for hemolysis in Palestinian children (81%), providing support for newborn G6PD deficiency screening programs. We report unanticipated molecular heterogeneity of G6PD variants among Gaza Strip Palestinians greater than reported in neighboring Arab populations. We report a high proportion of affected children with G6PD Cairo, which was observed previously in only a single Egyptian, and a novel mutation G6PD "Gaza".

Evaluation of three chitin metal silicate co-precipitates as a potential multifunctional single excipient in tablet formulations.

The performance of the novel chitin metal silicate (CMS) co-precipitates as a single multifunctional excipient in tablet formulation using direct compression and wet granulation methods is evaluated. The neutral, acidic, and basic drugs Spironolactone (SPL), ibuprofen (IBU) and metronidazole (MET), respectively, were used as model drugs. Commercial Aldactone, Fleximex and Dumazole tablets containing SPL, IBU and MET, respectively, and tablets made using Avicel 200, were used in the study for comparison purposes. Tablets of acceptable crushing strength (>40 N) were obtained using CMS. The friability values for all tablets were well below the maximum 1% USP tolerance limit. CMS produced superdisintegrating tablets (disintegration time < 1 min) with the three model drugs. Regarding the dissolution rate, the sequence was as follow: CMS > Fleximex > Avicel 200, CMS > Avicel 200 > Dumazole and Aldactone > Avicel 200 > CMS for IBU, MET and SPL, respectively. Compressional properties of formulations were analyzed using density measurements and the compression Kawakita equation as assessment parameters. On the basis of DSC results, CMS co precipitates were found to be compatible with the tested drugs. Conclusively, the CMS co-precipitates have the potential to be used as filler, binder, and superdisintegrant, all-in-one, in the design of tablets by the direct compression as well as wet granulation methods.

Possible association of 3' UTR +357 A>G, IVS11-nt 93 T>C, c.1311 C>T polymorphism with G6PD deficiency.

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.

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.

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.

Entamoeba histolytica or Entamoeba dispar among children in Gaza, Gaza Strip?

Most physicians in Gaza prescribe medicaments for patient's suffering from Entamoeba histolytica/dispar without parasitologic diagnosis. Additionally, stool analysis performed by the routine methods usually reports the species as E. histolytica without con-firmation. In this study, 92 stool specimens were collected and analyzed by wet mount, iron haematoxylin staining, antigen detection of E. histolytica and polymerase chain reaction (PCR). The total number of E. histolytica identified by PCR was 64 (69.6%) that of E. dispar was 21 (22.8%). Mixed infection with both E. histolytica and E. dispar was evident in 7 specimens (7.6%). In the light of these results approximately 30% of suspected clinical amoebiasis cases were negative for E. histolytica. It is recommended to use PCR for diagnosis of stool specimens from patients with E. histolytica/dispar and that treatment should be prescribed for only patients positive for E.