Ammar Madania

Characterization of mutations causing rifampicin and isoniazid resistance of Mycobacterium tuberculosis in Syria.

A cross sectional prospective study was carried out over a period of one year from (April 2008-March 2009 ), depending on prestructured questionnaire, study was carried out on a total of two hundred patients with different age, sex and with clinically suspected cases with tinea capitis and corporis. Specimens were obtained from skin scales of the lesion. Hair specimens were collected by plucking the hair with forceps. The aim of this study to identify the etiological agents involved in these infections. Out of 200 patients who presented with suspected superficial fungal and to determine prevalence of tinea capitis and tinea corporis in Benghazi population. Infected, 113 (56.5%) were male and 87 (43.5%) were female. Out of these, 117 children (65 male and 52 female) were provisionally diagnosed with tinea capitis and corporis. The youngest patient was a 5 months old infant, whereas the eldest patient a 71 year old man. Greater number of positive cases of dermatophytes is seen in children under the age of 15 year. Tinea capitis was predominant in 31 (57.4%) children, while tinea corporis were (14.8%) children. 125 (62.5%), were found to be positive by direct microscopic examination only, while 50 (25%) by culture only and 45 (22.5%) positive by both techniques. In addition 36 (18%) patients give positive family history of dermatophytosis, 9 patients of them were positive culture while 55 (27.5%) patients had history of contact with animals 16 of them were positive culture. Also17 (8.5%) were foreign patients, of these 8 were soudanense. In this study, the most common sites where dermatophytes in Tinea corporis isolated were the neck and back. Also we observed that, T. violaceum was the most common dermatophyte isolated 13 (24%) (mainly among children under age of 15 years). T. soudanense 9 (16%) was the second common isolated, followed by T. schoenleinii 8 (14.8%), other dermatophytes in descending order, were M. canis 5 (9.3%), T. mentagrophytes 4 (7.4%), M. ferrugineum 3 (5.5%), T. rubrum 3 (5.5%), T. tonsurans 2 (3.7%), M. nanum 2 (3.7%), T. yaoundi 1 (1.8%), T. terrestre 1 (1.8%), T. verrucosum 1 (1.8%), M. audouinii 1 (1.8%) and 1 (1.8%) were unidentified. Culture the isolates were a mixed of dermatophytes, in 2 cases of tinea capitis the culture revealed in mixed of T. violaceum and T. mentagrophytes, while 2 cases of tinea corporis; T. tonsurans and T. schoenleinii where the culture revealed a growth of T. rubrum and M. nanum. The infection was found to occur more frequently in males (29 cases than in females (25 cases). In the present study, grey patch was the predominant type of tinea capitis 32 (16%), black dot 2 (1%) and kerion 2 (1%) was the least common types.W investigated the anti-influenza virus activity of Acacia nilotica and possible mechanisms of action in vitro. We found that Acacia nilotica has anti-influenza-virus activity and both pre-incubation of virus prior to infection and post-exposure of infected cells with Acacia nilotica extract significantly inhibited virus yields. Influenza-virus-induced hemagglutination of chicken red blood cells was inhibited by Acacia extract treatment, suggesting that Acacia can inhibit influenza A virus infection by interacting with the viral hemagglutinin. Furthermore, Acacia extract significantly affect nuclear transport of viral nucleoprotein (NP). To best of our knowledge, this study revealed for the first time that Acacia nilotica extract can inhibit both viral attachment and replication and offers new insights into its underlying mechanisms of antiviral action. The fruit husk of Acacia nilotica collected from Sudan and extracted with 70% methanol. The crude extract was screened for its cytotoxicity against MDCK cell line by alamarBlue assay and WST-1 assay. Antiviral properties of the plant extract were determined by cytopathic effect inhibition assay and virus yield reduction assay (plaque assay). Time of addition assay and nuclear export mechanism were also performed.

Combination of conventional multiplex PCR and quantitative real-time PCR detects large rearrangements in the dystrophin gene in 59% of Syrian DMD/BMD patients

OBJECTIVES Adaptation of a low-cost protocol to diagnose large rearrangements of the dystrophin gene in DMD/BMD Syrian patients and to establish the distribution of these mutations in the 2 hotspots. DESIGN AND METHODS gDNA from 51 unrelated Syrian DMD/BMD male patients was isolated and analyzed by multiplex PCR of 25 hotspot exons in order to detect deletions. Patients who did not show any deletions were further analyzed by quantitative real-time PCR and the DeltaDeltaCt method in order to detect duplications in exons 4, 17, 47 and 52. RESULTS We found a deletion in 25 (49%) out of 51 patients studied. Quantitative real-time PCR revealed a duplication in 5 (9.8%) out of 51 patients. Combination of traditional multiplex PCR of hotspot exons with real-time PCR quantification of only exons 4, 17, 47 and 52 positively diagnosed 59% of Syrian DMD/BMD patients. CONCLUSION Our method may be useful as a cost-effective first-line test for the diagnosis of DMD/BMD patients before using exhaustive and expensive methods.

Effects of γ-radiation on cell growth, cell cycle and promoter methylation of 22 cell cycle genes in the 1321NI astrocytoma cell line

PURPOSE DNA damage caused by radiation initiates biological responses affecting cell fate. DNA methylation regulates gene expression and modulates DNA damage pathways. Alterations in the methylation profiles of cell cycle regulating genes may control cell response to radiation. In this study we investigated the effect of ionizing radiation on the methylation levels of 22 cell cycle regulating genes in correlation with gene expression in 1321NI astrocytoma cell line. METHODS 1321NI cells were irradiated with 2, 5 or 10Gy doses then analyzed after 24, 48 and 72h for cell viability using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliu bromide) assay. Flow cytometry were used to study the effect of 10Gy irradiation on cell cycle. EpiTect Methyl II PCR Array was used to identify differentially methylated genes in irradiated cells. Changes in gene expression was determined by qPCR. Azacytidine treatment was used to determine whether DNA methylation affectes gene expression. RESULTS Our results showed that irradiation decreased cell viability and caused cell cycle arrest at G2/M. Out of 22 genes tested, only CCNF and RAD9A showed some increase in DNA methylation (3.59% and 3.62%, respectively) after 10Gy irradiation, and this increase coincided with downregulation of both genes (by 4 and 2 fold, respectively). TREATMENT with azacytidine confirmed that expression of CCNF and RAD9A genes was regulated by methylation. CONCLUSIONS 1321NI cell line is highly radioresistant and that irradiation of these cells with a 10Gy dose increases DNA methylation of CCNF and RAD9A genes. This dose down-regulates these genes, favoring G2/M arrest.

Frequency of HLA-A alleles in the Syrian population genotyped by sequence-based typing

HLA‐A molecules are highly polymorphic. Their accurate typing at a high‐resolution level is crucial for successful organ, bone marrow and cord blood transplantation. Furthermore, several HLA alleles have been involved in susceptibility to autoimmune diseases, allergies, cancers and inflammations. In order to determine common HLA‐A alleles in Syria and their frequencies, sequence‐based typing (SBT) was used to genotype HLA‐A alleles at high resolution (four digit level) among one hundred and thirty randomly selected Syrian individuals. Exons 2, 3 and 4 of the HLA‐A gene were amplified by PCR and sequenced. The sbt‐engine software was used for allele assignment. Ambiguities were solved using group‐specific sequencing primers (GSSPs). We could identify 32 different HLA‐A alleles which were divided into 3 groups: high frequency (approximately 10%, A*01:01; A*24:02; A*03:01; A*02:01), moderate frequency (approximately 3%, such as A*02:05, A*31:01 and A*33:01), and low frequency (approximately 1%, such as A*02:11, A*29:01, A*02:02 and A*36:01). Homozygosity rate was higher than expected (11.5% vs. 7.15%). For high frequency alleles, our results show similarity to neighbouring countries. However, 15 alleles (such as A*02:04, A*02:06, A*02:11 and A*02:17) found in our cohort in low frequencies were never reported in some or all neighbouring countries. This is the first report on HLA‐A allele frequencies in Syria. In spite of the relatively low number of tested subjects, our results revealed a high degree of diversity, with 32 different alleles, reflecting the high ethnic heterogeneity of the Syrian population. The identification of alleles rarely or never reported in neighbouring countries indicates a higher genetic diversity in Syria.

Effects of fenofibrate on Semaphorin 6B gene expression in rat skeletal muscle

Semaphorin 6B is a member of the semaphorin family of signaling proteins, which is implicated in a variety of biological processes, such as axon guidance and muscle regeneration. PPARα is known to be a regulator of semaphorin 6B in human cancer cell lines. In this study, we examined the effect of fenofibrate as a PPARα activator on the expression of the Sema6B gene in rat skeletal muscle. A total of 18 rats were divided into two groups: one fed with standard chow (control group), and the other with fenofibrate chow (treatment group) for 4 weeks by gavage with 30 mg/kg per day. The expression of rSema6B mRNA was analyzed by qPCR. rSema6B protein content in rat skeletal muscle was measured by Western blotting. A significant down-regulation of rSema6B was observed at both the mRNA and protein levels. Using the ChIP assay, a putative PPARα binding region (PPRE) was identified in the rSema6B promoter. We conclude that the expression of Sema6B is down-regulated by fenofibarte in rat skeletal muscle.

Mild androgen insensitivity syndrome (MAIS): the identification of c.1783C>T mutation in two unrelated infertile men

Two unrelated men complaining of primary male infertility presented to Orient Hospital in Damascus city. Physical examination showed moderate hypoandrogenic features. Both men were azoospermic. Hormone profiles revealed an elevation of follicle-stimulating hormone in one patient, but all the other hormones tested were within normal limits for both patients. Further genetic analyses, including karyotype and microdeletions in the AZF region of the Y chromosome, were normal in both patients. Mild androgen insensitivity syndrome was expected in the two patients. Sequencing analysis of the first exon in the androgen receptor (AR) gene have shown c.1783C>T mutation in the two patients with azoospermia. This paper sheds light on the need to screen for mutations in the AR gene, causing male infertility whenever mild hypoandrogenic features are present with unexplained male infertility.