SNP-PCR genotyping links alterations in the GABAA receptor (GABRG3: rs208129) and RELN (rs73670) genes to autism spectrum disorder among peadiatric Iraqi Arabs.

INTRODUCTION: Autism spectrum disorder (ASD) is an increasing concern among the Iraqi Arab population. The genetic alterations that cause ASD are likely to converge at the synapse. This study investigated polymorphisms in the GABAA receptor subunit (GABRG3) and the RELN gene as putative biomarkers of ASD in a pediatric population in Iraq. METHODS: The case control study included 60 patients with a clinical diagnosis of ASD (mild, moderate, or severe) according to DSM-IV criteria and matched healthy controls (n = 60). Blood samples were collected for DNA genotyping of SNPs rs736707 and rs208129 for RELN and GABRG3 using allele specific PCR. Assessment of genotype and allele distributions in patient groups used odd ratios (OR) with 95% confidence intervals and the Chi-square test. All statistical analysis was performed used SPSS software. RESULT: The patient cohort was highly consanguineous, with increased ratio (p > 0.05) of males to females (3:1) in both ASD (mean age, 6.66 ± 3.05) and controls (mean age, 5.76 ± 2.3). Both GABRG3 rs208129 genotypes TT (OR 4.33, p = 0.0015) and TA (OR 0.259, P = 0.008), and the T and A alleles were significantly associated with ASD. The RELN rs736707 TC genotype (OR 2.626, P = 0.034) was the only significant association with ASD. CONCLUSION: GABRG3 SNP rs208129 is a leading biomarker to predict genetic vulnerability to ASD in Iraqi Arabs. Expanded SNP panels and increased sample sizes are required for future GABRG3 studies, and to reach a consensus on RELN utility. Future ASD screening programs in Iraq should include genetic metrics in addition to clinical phenotype assessments.

FISH comets show that the salvage enzyme TK1 contributes to gene-specific DNA repair.

Thymidine kinase 1 (TK1) is a salvage enzyme that phosphorylates thymidine, imported from surrounding fluids, to create dTMP, which is further phosphorylated to the DNA precursor dTTP. TK1 deficiency has for a long time been known to cause increased cellular sensitivity to DNA damage. We have examined preferential strand break repair of DNA domains in TK1(+) and TK1(-) clones of the Raji cell line, by the Comet-FISH technique, in bulk DNA and in the actively transcribed tumor suppressor (TP53) and human telomerase reverse transcriptase (hTERT) gene regions, over 1 h after 5Gy γ-irradiation. Results showed that repair of the TP53 and hTERT gene regions was more efficient in TK1(+) compared to TK1(-) cells, a trend also reflected to a lesser degree in genomic DNA repair between the cell-lines. The targeted gene-specific repair in TK(+) cells occurred rapidly, mainly over the first 15 min repair-period. Therefore, TK1 is needed for preferential repair of actively transcribed regions, through a previously unsuspected mechanism. In principle, TK1 could exert its protective effects through supply of a supplementary dTTP pool for accurate repair of damaged genes; but Raji TK1(+) cells in thymidine free media still show preferential repair of transcribed regions. TK1 therefore does not exert its protective effects through dTTP pools, but through another unidentified mechanism, which affects sensitivity to and mutagenicity by DNA damaging agents.

Specific-mutational patterns of p53 gene in bladder transitional cell carcinoma among a group of Iraqi patients exposed to war environmental hazards.

BACKGROUND: To unfold specific-mutational patterns in TP53 gene due to exposures to war environmental hazards and to detect the association of TP53 gene alteration with the depth of bladder cancer. METHODS: Twenty-nine bladder carcinomas were analyzed for TP53 alterations. PCR-single strand conformational polymorphism analysis, DNA sequencing and immunohistochemical analysis using monoclonal mouse anti-human p53 antibody (Clone DO-7) were employed. RESULTS: TP53 gene mutations occurred in 37.9% of the cases while TP53 overexpression occurred in 58.6%. Both of them were associated with deep invasive-tumors. Single mutations were seen in 63.6%, whereas only 27.3% have shown double mutations. Four mutations were frameshifted (30.8%); two of them showed insertion A after codon 244. There was no significant association between TP53 mutations and protein overexpression (P>0.05), while a significant association was observed between TP53 alterations and tumors progression (P ≤ 0.01). CONCLUSION: The infrequent TP53mutations, especially insertion A and 196 hotspot codon, may represent the specific-mutational patterns in bladder carcinoma among the Iraqi patients who were exposed to war environmental hazards. TP53 alteration associated with bladder cancer progression should be analyzed by both mutational and protein expression analysis.

Exposure to depleted uranium does not alter the co-expression of HER-2/neu and p53 in breast cancer patients.

BACKGROUND: Amongst the extensive literature on immunohistochemical profile of breast cancer, very little is found on populations exposed to a potential risk factor such as depleted uranium. This study looked at the immunohistochemical expression of HER-2/neu (c-erbB2) and p53 in different histological types of breast cancer found in the middle Euphrates region of Iraq, where the population has been exposed to high levels of depleted uranium. FINDINGS: The present investigation was performed over a period starting from September 2008 to April 2009. Formalin-fixed, paraffin-embedded blocks from 70 patients with breast cancer (62 ductal and 8 lobular carcinoma) were included in this study. A group of 25 patients with fibroadenoma was included as a comparative group, and 20 samples of normal breast tissue sections were used as controls. Labeled streptavidin-biotin (LSAB+) complex method was employed for immunohistochemical detection of HER-2/neu and p53.The detection rate of HER-2/neu and p53 immunohistochemical expression were 47.14% and 35.71% respectively in malignant tumors; expression was negative in the comparative and control groups (p < 0.05).HER-2/neu immunostaining was significantly associated with histological type, tumor size, nodal involvement, and recurrence of breast carcinoma (p < 0.05), p53 immunostaining was significantly associated with tumor size, nodal involvement and recurrence of breast cancer (p < 0.05). There was greater immunoexpression of HER-2/neu in breast cancer in this population, compared with findings in other populations.Both biomarkers were positively correlated with each other. Furthermore, all the cases that co-expressed both HER-2/neu and p53 showed the most unfavorable biopathological profile. CONCLUSION: P53 and HER-2/neu over-expression play an important role in pathogenesis of breast carcinoma. The findings indicate that in regions exposed to high levels of depleted uranium, although p53 and HER-2/neu overexpression are both high, correlation of their expression with age, grade, tumor size, recurrence and lymph node involvement is similar to studies that have been conducted on populations not exposed to depleted uranium. HER-2/neu expression in breast cancer was higher in this population, compared with results on non-exposed populations.

Vascular endothelial growth factor (VEGF) receptor expression correlates with histologic grade and stage of colorectal cancer.

BACKGROUND: Colorectal carcinoma (CRC) is the seventh-most common malignancy and is the main cause of death in Iraq. The incidence of this cancer has increased sharply after the invasion of Iraq in 2003. AIM: To estimate immunohistochemical expression of vascular endothelial growth factor (VEGF) in CRC in relation to other parameters, such as grade and stage of tumour. METHODS: Formalin fixed, paraffin-embedded blocks from 52 patients (27 male and 25 female) with CRC were included in this study. A group of 22 patients with non-cancerous colonic tissues were included as a control group. Avidin-biotin complex method was employed for immunohistochemical detection of VEGF. RESULTS: VEGF immuno-expression was positive in 51.9% of CRC, while it was 18.2% in the normal colonic tissue (p <0.05). VEGF immunostaining was positively correlated with grade of colonic malignancy (p <0.05). CONCLUSION: These findings provide further evidence for the role of VEGF in the carcinogenesis of CRC. However, VEGF could not be well correlated with stage of tumour and hence may be a poor prognostic parameter of state of malignancy of colonic carcinoma.

Nanoceria have no genotoxic effect on human lens epithelial cells.

There are no treatments for reversing or halting cataract, a disease of the structural proteins in the eye lens, that has associations with other age-related degenerative conditions such as Alzheimer's disease. The incidence of cataract and associated conditions is increasing as the average age of the population rises. Protein folding diseases are difficult to assess in vivo as proteins and their age-related changes are assessed after extraction. Nanotechnology can be used to investigate protein changes in the intact lens as well as for a potential means of drug delivery. Nanoparticles, such as cerium oxide (CeO(2)) which have antioxidant properties, may even be used as a means of treating cataract directly. Prior to use in treatments, nanoparticle genotoxicity must be tested to assess the extent of any DNA or chromosomal damage. Sister chromatid exchanges were measured and DNA damage investigated using the alkaline COMET assay on cultured human lens epithelial cells, exposed to 5 and 10 microg ml(-1) of CeO(2) nanoparticles (nanoceria). Nanoceria at these dosages did not cause any DNA damage or significant increases in the number of sister chromatid exchanges. The absence of genotoxic effects on lens cells suggests that nanoceria, in the doses and exposures tested in this study, are not deleterious to the eye lens and have the potential for use in studying structural alterations, in developing non-surgical cataract treatments and in investigating other protein folding diseases.

Expression of VEGF in urinary bladder transitional cell carcinoma in an Iraqi population subjected to depleted uranium: an immunohistochemical study.

The present study aimed to assess the correlation between vascular endothelial growth factor (VEGF) overexpression and the grade, size, and recurrence of transitional cell carcinoma (TCC) in the south of Iraq, which includes regions that have been exposed to high levels of depleted uranium. The study also sought to evaluate whether there is any biomarker in the expression that could be correlated with the increased incidence of this type of cancer in the exposed areas. Samples of formalin-fixed and paraffin-embedded tissue from 54 patients (41 males and 13 females) with TCC and from 32 patients with benign bladder lesions (cystitis) used as controls were included in this study. The avidin-biotin complex method was used for immunohistochemical detection of VEGF. VEGF immunoexpression was positive in 77.77% of TCC but was not found in benign bladder lesions (cystitis) (P<0.05). VEGF immunostaining was positively correlated with grade, stage, and recurrence of TCC but the findings were not statistically significant (P>0.05). These findings support the role of VEGF in the carcinogenesis of TCC regarding evolution, behavior, and aggressiveness. Hence, VEGF could be considered as a poor prognostic parameter in bladder cancer. No positive correlation between immunohistochemical expression and the high incidence of TCC was detected (R=<0.3). The study further concludes that immunohistochemical expression of the VEGF gene in TCC bladder cancer does not differ from similar cancers found in other parts of the world where there has been no known exposure to depleted uranium.

High prevalence of HER-2/neu overexpression in female breast cancer among an Iraqi population exposed to depleted uranium.

BACKGROUND: This study aimed to estimate the rate of HER-2/neu (c-erbB2) immunohistochemical overexpression in different histological types of breast cancer found in the middle Euphrates region of Iraq, a region that was exposed to high levels of depleted uranium. HER-2/neu (c-erbB2) overexpression was correlated with common clinicopathological parameters such as age, grade, stage, tumor size and lymph node involvement to determine if any particular biomarker for exposure to depleted uranium could be found in the tumor samples from this region. MATERIALS AND METHODS: The present investigation was performed over a period starting from September 2007 to June 2008. Formalin-fixed, paraffin-embedded blocks from 90 patients with breast cancer were included in this study. A group of 25 patients with benign breast lesions (fibroadenoma) was included as a comparative group, and 20 breast tissue sections were used as controls. Labeled streptavidin-biotin (LSAB) complex method was employed for immunohistochemical detection of HER-2/neu. RESULTS: HER-2/neu immuno-expression was positive in 67.8% of breast cancer, while it was negative in all benign breast lesions (fibroadenoma) ( P P P > 0.05). CONCLUSION: Based upon the findings of this study, it can be concluded that HER-2/neu overexpression plays an important role in the pathogenesis of breast cancer and is associated with a worse prognosis. The findings indicate that in regions exposed to high levels of depleted uranium, HER-2/neu overexpression is high, but its correlation with age, grade, stage, tumor size, and lymph node involvement is similar to studies that have been conducted on populations not exposed to depleted uranium.

Over expression of vascular endothelial growth factor in correlation to Ki-67, grade, and stage of breast cancer.

OBJECTIVE: To assess the significance of vascular endothelial growth factor (VEGF) protein over expression in human breast cancer, and its possible correlation with cell proliferation marker (Ki-67), grade and stage of breast cancer. METHODS: We carried out this study at the Department of Pathology, Kufa University, between November 2006 and September 2007. A retrospective study was employed on paraffin-embedded blocks from 52 female patients with breast cancer. A group of 21 patients with benign breast lesions was included for comparison and 14 cases of normal breast tissue as a control group. This investigation designed to employ immunohistochemistry using Avidin-Biotin Complex (ABC) method for detection of both VEGF and Ki-67. RESULTS: A total of 87 samples was included. Vascular endothelial growth factor immunoexpression was considered as positive in 61.5% of malignant and in 19% of benign breast lesions. No over expression sign has been noticed in normal breast tissue (p<0.005). No significant difference in VEGF over expression among different histological types of breast cancer (p>0.05). Vascular endothelial growth factor immunostaining was positively correlated with Ki-67, grade, stage, lymph node metastasis, and recurrence of breast cancer (p<0.05). No such correlation has been seen when the age of the patients has been considered. CONCLUSION: Vascular endothelial growth factor Vascular endothelial growth factor plays an important role in pathogenesis of breast cancer evolution, and supports the evidence of its role in angiogenesis and cell survival. This study recommended that the blocking of VEGF may be a target for blocking angiogenesis and hence improving the efficacy of anti-cancer therapy.

The role of Y chromosome C-band size polymorphism in male infertility with a reference to their effect on the total length of the chromosome.

OBJECTIVE: To assess the contribution of chromosome on sperm production. METHODS: During a 2 year period from October 1999 to December 2001 a total of 200 male patients were included in the present study. The study was carried out at Kufa Medical College, Kufa, Iraq. Blood culture, chromosomal harvesting, Giemsa stain, C-band technique and C-band size calculation have been conducted according to the standard methods. RESULTS: No statistical differences have been recorded in the C-band size distribution level among the infertile groups compared to normal control. The functional relationship between C-band, euchromatin and the total length of the Y chromosome showed a strong correlation coefficient among the infertile groups (oligospermia, azoospermia) as well as the normal control. CONCLUSION: In our qualitative study the C-band size of the Y chromosome has no effect on spermatogenesis. The decrease in the size of Y chromosome is due to the decrease in both the heterochromatin and euchromatin regions.

Total cellular ribose nucleic acid analysis of multi-drug resistant cancer cells.

OBJECTIVE: The present study is designed to elucidate the correlation between gene dosage and increased messenger ribose nucleic acid (RNA) level in multi-drug resistant cancer cells. METHODS: The human lymphoblasts CCRF-CEM (CEM) and CEM-vinblastine (VBL) 10, CEM-VBL 20, CEM-VBL 40, CEM-VBL 60, cell line CEM-VBL 80, and CEM-VBL 100 were derived from CEM VBL 10 by single step selection technique. For the analysis of total RNA and deoxyribonucleic acid (DNA), both Northern-Southern blot analysis were carried out on all sensitive and resistant cell lines. Part of the study was conducted at the Immunology Laboratory, Higher Institute of Health, Rome and the other part was conducted at the Middle Euphrates Center for Cancer Researches, Kufa University, Iraq, between 1990 and 2000. RESULTS: Total cellular RNA was analyzed to quantitate the levels of expression of multi-drug resistant-one gene with extent of its amplification in CEM-sensitive and resistant cell lines. Only the CEM-VBL resistant over expresion the multi-drug resistant-one gene but in CEM-sensitive cells do not. The over-expresses appears to correlate proportionally with the level of drug resistance. No such correlation has been detected with regard to the genomic DNA. CONCLUSION: It appears that the first step in the transition from drug sensitive to drug resistant is increasing levels of messenger RNA followed by gene amplification. Furthermore, once the gene is amplified, it remains at the same level of amplification regardless of the concentration of drug in which given cells have been selected.

High frequency of satellite association in metaphases of infertile male patients.

OBJECTIVE: The present study is designed to evaluate the contribution of satellite association phenomenon on spermatogenic impairment. METHODS: During a one year period, January 1999 through to January 2000, the frequency of satellite association has been investigated in 57 patients, with the clinical diagnosis of male infertility. This study was carried out at the Middle Euphrates Center for Cancer Researches, College of Medicine, Kufa University, Kufa, Iraq. Of those 57 patients, 33 patients were diagnosed with azoospermia and 24 patients with oligospermia, the efficacy evaluation was based on the measurement of 8 satellite association parameters. Blood culture and chromosome harvesting was carried out according to our standard methods. RESULTS: Our experience of association behavior of acrocentric chromosome in the 57 infertile males showed statistically significant difference in infertile male classes compared to control groups. Eight studied parameters were included in the evaluation (P<0.005). CONCLUSION: The significant increase in the satellite association is proposed to have another indirect causal factor, which influenced spermatogenesis. Furthermore, the satellite association technique may be used as a laboratory method for the evaluation of male infertility.

The significance of autosomal C-band size polymorphism in male infertility.

OBJECTIVE: The aim of this study is to assess the possible role of autosomal C-band size polymorphism in male infertility. METHODS: Two-hundred male patients with clinical diagnosis of infertility and 100 normal controls were included in the present investigation. All patients were assessed by Urologist Consultant at the Department of Pathology and Forensic Medicine, Kufa University, Kufa, Iraq, during a 2-year-period, October 1999 to October 2001. C-band evaluation was based on both quantitative and qualitative methods. Blood culture, chromosome harvesting, and C-band technique were carried out according to standard methods. RESULTS: 1. C-band quantitative study indicates a significant increase in the C- band size of chromosomes 9 and 16 among infertile groups as compared to normal fertile group (p<0.01). 2. C-band qualitative study indicates a significant increase in the C-band size (level 3) of chromosome number one among the infertile group as compared to normal fertile group (p<0.01). CONCLUSION: The present findings require further extensive study to shed light on the possible correlation between C-band polymorphism and male infertility.