Adenomatous polyposis coli gene large deletions in Iranian patients with familial adenomatous polyposis.

Context: Familial adenomatous polyposis (FAP) is one type of hereditary colon cancer with a large number of precancerous polyps that initiation to growth in childhood and adolescent. Mutation in adenomatous polyposis coli (APC) gene is the cause of FAP. Aims: The aim of the current study was to standardize multiplex ligation probe amplification (MLPA) method in screening of APC large deletions for the first time in Iranian patients with FAP. Subjects and Methods: Deoxyribonucleic acid was extracted from 34 FAP patients by saluting out method. All patients were screened for APC large deletions whit MLPA and for the positive results, respective region was investigated by polymerase chain reaction sequencing. All genetic alterations were doubled checked in two separated rounds of MLPA. Results: The detection rate of large fragment deletions in APC was 5.8% (2/34). Both of the Iranian patients had deletion in the middle and the end of exon 15, however, comparing of clinical features between patient with the large deletion and patients without deletion did not show any significant difference in each variable including, age at diagnosis, signs of disease and poly type. Conclusions: It seems that exon 15 of APC gene is probably the hotspot region in Iranian FAP patients. Association of genotype/phenotype is well known in FAP patients, but in this study statistical analyses did not show a significant difference in each considerable factor between patients with and without large deletions. It seems better to consider MLPA as an initial step to screening APC mutations.

[One novel PMS2 germline mutation in a patient with hereditary non polyposis colorectal cancer]

Hereditary non polyposis colorectal cancer [HNPCC] is an autosomal dominant syndrome. The cancer appears between 40 and 50 years of age. Mutation in mismatch repair genes can lead to this cancer .One of the genes which is involved in this disease is PMS2 gene. Here, we present a case with a novel germline mutation in PMS2 gene. The aim of this report was to examine PMS2 gene and identify novel germline mutations in this gene. A 77-year-old male with diagnosis of colorectal carcinoma was referred for genetic testing. He suffered from a polyp with a diameter of 6.8 cm in hepatic flexure. The patient did not meet Amsterdam Criteria and Bethesda Guidelines, but screening for HNPCC was performed on account of pathological findings. Blood sample was used for identification of mutation and the paraffin embedded block was prepared for MSI analysis. One mutation in PMS2 gene was detected by analysis of the amplicon sequencing. The mutation was a transitional mutation in position 676 which led to transformation of guanine to adenine resulting in substitution of glutamic acid for glycine. Immunohistochemistry confirmed abnormal expression of PMS2 gene and MSI assay showed instability of sequenced amplicons in this gene

[Study of three common mitochondrial mutations in Arab patients with nonsyndromic hearing loss in Khuzestan province, I.R.Iran]

Hearing Impairment [HI] is the most prevalent neurosensory disorder occurs in 1/1000 newborn. The majority of hearing deficiencies are of genetic origin. About%0-2 of the genetic HI cases are due to mutations in mitochondrial genes. In the present study we investigated the frequency of 3 mtDNA A1555G, A3243G and A7445G mutation of 62 patients with nonsyndromic hearing loss in Khuzestan province. In this descriptive study, we investigated the presence of three mitochondrial mutations; A1555G, A3243G and A7445G in 62 Arab subjects with autosomal recessive non syndromic hearing loss in Khuzestan province. DNA was extracted using standard phenol -chloroform method. The screening of the mitochondrial gene mutations was performed by PCR-RFLP procedure.The possible mutations were confirmed by direct sequencing. None of the investigated mutations; A1555G, A3243G and A7445G were detected in this study. However PCR-RFLP revealed two mutations; G3316A, A7445C in 2 deaf subjects studied. This study is shown that mtDNA mutations consist of G3316A and A7445C are responsible for few of ARNSHL in sample studied and none of the A1555G, A3243G and A7445G mutations are responsible for ARNSHL in this population. The data presented here will improve the genetic counseling of hearing impaired patients in Khuzestan province

[Frequency of the common mitochondrial DNA [mtDNA] mutations in non-syndromic hearing impairment in southwest subpopulations of Iran]

Although, most hereditary non-syndromic hearing impairment [NSHI] is due to mutation in nuclear genes, role of mtDNA mutations in causing deafness becoming much more clear in recent years. The aim of the present study was to screen the A1555G, C1494T, A3243G and A7445G mutations in non-syndromic hearing impairment patients in two provinces of southwest of Iran. In this descriptive laboratory study, 150 subjects with acquired and prelingual autosomal recessive NSHI from Chaharmahal va Bakhtiari province and 46 unrelated probands with postlingual NSHI from Bushehr province [negative for GJB2 mutations] were screened for the presence of the common mtDNA mutations using PCR-RFLP method that followed by direct sequencing for confirming the observed mtDNA mutations. None of these mutations was found in subjects with acquired and prelingual autosomal recessive NSHI from Chaharmahal va Bakhtiari province, but mutation A1555G with frequency of 4.35% was found in postlingual NSHI patients in Bushehr province. This investigation shows that apparently, mtDNA mutations play a more significant role role in the etiology of postlingual NSHI in comparison with prelingual NSHI

Application of nanofiltration membrane in the separation of amoxicillin from pharmaceutical wastewater

Separation of amoxicillin from pharmaceutical wastewater by nanofiltration [NF] membrane has been investigated in this study. For this purpose a membrane system including a polyamide spiral wound NF membrane was evaluated for the treatment of amoxicillin wastewater. The effects of operating conditions such as flow rate, pressure and concentration of amoxicillin and COD in the feed, on the efficiency of the membrane were evaluated. The permeation flux and rejection of amoxicillin and COD were the criteria for this evaluation. The rejection of the amoxicillin by the selected NF membrane was adequate and in most cases exceeded 97% whereas COD reached a maximum of 40% rejection and permeation flux was over 1.5 L/min.m[2]. The rise in pressure enhanced the transport rate of the solvents. Permeation flux of the NF membrane increased with increasing flow rates. Experimental data also indicated that concentration polarization existed in this membrane separation process. The stable permeation flux and high rejection of amoxicillin indicated the potential of NF for the recovery of amoxicillin from the pharmaceutical wastewater

Genetic linkage analysis of 15 DFNB loci in a group of Iranian families with autosomal recessive hearing loss

Hearing loss [HL] is the most frequent sensory birth defect in humans. Autosomal recessive non-syndromic HL [ARNSHL] is the most common type of hereditary HL. It is extremely heterogeneous and over 70 loci [known as DFNB] have been identified. This study was launched to determine the relative contribution of more frequent loci in a cohort of ARNSHL families. Thirty-seven Iranian families including 36 ARNSHL families and 1 family with Pendred syndrome each with >/= 4 affected individuals, from seven provinces of Iran, were ascertained. DFNB1 contribution was initially studied by DNA sequencing of GJB2 and linkage analysis using the relative STR markers. The excluded families were then subjected to homozygosity mapping for fifteen ARNSHL loci. Sixteen families were found to be linked to seven different known loci, including DFNB1 [6 families], DFNB4 [3 families +1 family with Pendred syndrome], DFNB63 [2 families], DFNB2 [1 family], DFNB7/11 [1 family], DFNB9 [1 family] and DFNB21 [1 family]. DNA sequencing of the corresponding genes is in progress to identify the pathogenic mutations. The genetic causes were clarified in 43.2% of the studied families, giving an overview of the causes of ARNSHL in Iran. DFNB4 is ranked second after DFNB1 in the studied cohort. More genetic and epigenetic investigations will have to be done to reveal the causes in the remaining families

[Screening of mitochondrial mutations of A1555G, A 3243G, and A7445G in MTRNR1, MTTL1 and MTTS1 genes in subjects with nonsyndromic sensorineural hearing loss]

Various frequencies of the mtDNA mutations have been reported from different population world wild. Three mitochondrial DNA [mtDNA] mutations including A1555G, A 3243G, and A7445G which occurred in MTRNR1, MTTL1 and MTTS1 genes were considered as the main causes of mitochondrial hearing loss in some populations. To determine the frequency of the A1555G, A3243G, and A7445G mutations in nonsyndromic sensorineural hearing loss subjects in Gilan. Forty six subjects with nonsyndromic sensorineural hearing loss were screened by provided questionnaire and audiogram from Gillan Welfare Organization. PCR-RFLP procedure was used in order to presence the MtDNA of A1555GA 3243G and A7445G mutations and was confirmed by subsequent direct sequencing. There was no MtDNA of A1555G, A3243G and A7445G mutation in the cohort study of 46 deaf individuals. Investigation of PCR-RFLP of the MTTL1 gene for existence A3243G mutation lead to identification a G3316A variant that destroyed other restriction site, in the other site of PCR fragment. Our finding indicated that possibility the association of mitochondrial mutations with deafness is very low in deaf subjects in north of Iran. According to existence the G3316A that its pathogenesis in relation to hearing loss phenotype has not stabilized, the frequency of G3316A is 1.46% that can be had highlights role of mitochondrial mutation in deafness

[Survey the association between tissue eosinophil counts and prognostic factors of esophageal SCC]

According to World Health Organization reports, the esophageal Squamous Cell Carcinoma [SCC] is most frequent in north-east of Iran. Infiltration of inflammatory cells, such as eosinophils, may play an important role in biologic behaviors of tumors. Survey the association between tissue eosinophil counts and prognostic factors of esophageal SCC. This analytical cross-sectional study was conducted applying a convenience sampling. The archive of pathology department of Imam-Reza Hospital, Mashhad, between 2005-2007 was standard method of Tumor-Associated Tissue Eosinophilia. Mean numbers of eosinophils and mastocytes between different groups of prevascular or preneural invasion, lymph node metastases, tissue differentiation status and depth of invasion were tested statistically. Twenty-five [61.0%] men and 16 [39.0%] women with a mean [Standard Deviation, SD] age of 53.6 [ +/- 14.78] were studied. Mean [SD] number of eosinophils and mastocytes were 115.4 [ +/- 88.16] and 30.7 [ +/- 26.03] respectively. Mean number of eosinophils was higher in patients with prevascular invasion [183.2 vs. 86.9] [p=0.002]. No other statistically significant association was found. Excess number of tissue eosinophils indicates the tendency towards more prevascular invasion. It is recommended to follow these patients intensively

[Molecular investigation of mtDNA A1555G, A3243G and A7445G mutations among the non syndromic hearing loss cases in Fars, Iran]

Hearing loss is a sensorineural disorder occuring in 1 out of 500 births. It happens due to some genetic/environmental causes or both. More than 60% of cases are noninherited and 80% non syndromic with autosomal recessive inheritance. In the present study we investigated the frequency of mtDNA A1555G, A3243 and A7445G mutations among the patients in Fars province. Seventy two non syndromic hearing loss subjects were studied. DNA was extracted using standard phenol-chloroform method. The screening of the mitochondrial gene mutations were performed using PCR-RFLP procedure. Finally, the possible mutations were confirmed by direct sequencing. None of the A1555G, A3243G and A7445G mutations was detected in this study. However, destroying a MTTL1 restriction site for the investigation of A3243G mutation, revealed a G3316A with allelic variant of 1.4% in the deaf subjects. Our data indicated that the mitochondrial A1555G, A3243 and A7445G mutations have no role in auditory deficits in patients studied

[Mutation screening of GJB2 and GJB6 and genetic linkage study of three prevalent DFNB loci in Iranian families with autosomal recessive non-syndromic hearing loss]

The incidence of prelingual hearing loss [HL] is about 1 in 1000 neonates of which, more than 60% of cases are inherited. Non-syndromic HL [NSHL] is extremely heterogeneous: more than 100 loci have been identified. The most common form of NSHL is the autosomal recessive form [ARNSHL]. Here, we have investigated CX26 [GJB2] and CX30 [GJB6] gene mutation and linkage analysis of 3 known loci in Iranian families. A cohort of 36 big ARNSHL pedigrees from 7 provinces of Iran was investigated. All of the families were examined for the presence of GJB2 and GJB6 [del D13S1830 and del D13S1854] mutations using direct sequencing and multiplex PCR, respectively. The negative mutations pedigrees for the above-mentioned mutations, were then tested for the linkage to the 3 known loci, including DFNB3[MYO7A], DFNB4[SLC26A4] and DFNB7/11[TMC1], using STR markers and conventional PCR and PAGE. Six families had GJB2 mutations. No GJB6 mutation was found. Totally, 3 families showed linkage to DFNB4 and 1 family was linked to DFNB7/11. DFNB1 [GJB2] and DFNB4 are the main causes of ARNSHL in our study samples and GJB6 mutations are apparently absent in the Iranian population

Association between genetic polymorphism of methylenetetrahydrofolate reductase with non familial Colorectal Cancer in Iranian population

Methylenetetrahydrofolate reductase [MTHFR] is a key enzyme regulating folate metabolism, which affects DNA methylation and synthesis. One of the most important polymorphisms identified in the MTHFR gene is C677T. MTHFR activity is lowered in individuals with 677TT genotype. Using pyrosequencing, we analyzed the MTHFR genotypes in 118 colorectal cancer patients and 189 normal matched controls. Whereas the CC, CT and TT genotypes of MTHFR among the colorectal cancer patients were 51.7%, 28.0% and 20.3% respectively, we were able to find 47.1% of 677CC, 27.0% of 677CT and 25.9% of 677TT in normal controls. An inverse association was observed between the risk of colorectal cancer and TT genotype with the odds ratios [OR] of 1, 0.94 and 0.71 for CC, CT, and TT genotypes, respectively. This association was similar in both sexes, but in patients with high levels of folate intake. Our study corroborates previous findings of an inverse association between MTHFR 677TT genotype and colorectal cancer, especially at high levels of folate

Association between polymorphism of MTHFR gene and non-familial colorectal cancer

Colorectal cancer is one of the most common malignancies in worldwide. Because the gene 5, 10-methylene-tetrahydrofolate reductase [MTHFR] plays a key role in methylation, synthesis and repair of DNA, numerous studies have focused on evaluating the correlation between polymorphisms of this gene and sporadic colorectal cancer. This study was carried out to examine the association of MTHFR gene polymorphism, C677T, with non-familial colorectal cancer in an Iranian population. We analyzed peripheral blood samples of 118 cases of colorectal cancer and 189 controls by pyrosequencing method. Controls were subjects who had been referred to our center during the study period and had revealed normal findings on colonoscopy. We found that frequency of CC, CT and TT genotypes among the colorectal cancer patients were 51.7%, 28% and 20.3% respectively. The figures for controls were 47.1%, 27% and 25.9% respectively. Furthermore, allele frequency T in the cases was 34% and allele frequency C was 66% while allele frequency T in controls was 39% and allele frequency C was 61%. Interestingly we observed a reverse association between risk of colon cancer with 677TT genotype

[Height, height and body mass index of 6-15-year-old school girls in Babol, northern Iran between 1995 to 2002: a comparison with the US 2000 centers for disease control and prevention references]

Since there are no national references for height, weight and body mass index [BMI] or Iranian children, the United States Centers for Disease Control and Prevention [CDC] 2000 standards are widely used in Iran. An assessment of the suitability of these standards was thought essential. The aim of this study was to compare the growth indices of 6-15-year-old school girls in Babol, Northern Iran, with the CDC 2000 standards. Six hundred and nine healthy 6-15-year-old Babol schoolgirls, selected by the convenience sampling method, were studied retrospectively between 1995 and 2002. The reference percentiles for height, weight and body mass index were calculated using the LMS method [the LMS Light 1.28 software]. After converting the growth data into standard deviation scores [SDS] based on the CDC2000 reference, the mean SDS within and through the age groups was compared with zero using the one sample T-test and SPSS 10.0.1. Statistically significant differences would mean significant differences between the growth indices of the population studied and the CDC2000 reference standards. The mean SDS for height, weight and BMI were -0.2 [SD - 1.00], -0.3 [SD =1.12] and -0.3 [SD =1.09], respectively, all significantly different from zero [p=0.000]. These scores reflect the magnitudes of differences from the CDC2000 references. These differences decreased as age increased and were more prominent in the 6- to 11 -year old girls. In these girls height, weight and BMI SDS were - 0.24 [SD =1.01], -0.48 [SD =1.10] and -0.46 [SD =1.15], respectively [p -0.000]. Growth charts were drawn. The height, weight and BMI values for the population studied were all lower than the corresponding CDC2000 standards. Therefore, the CDC standards are not suitable for use for Babol girls. Considering the nonprobability sampling method used in this study, any generalization should be made with caution. This study supports the idea of developing appropriate local standards for practical use

Frequencies of mutations in the connexin 26 gene [GJB2] in two populations of Iran [Tehran and Tabriz]

While hearing loss has been considered to be a very heterogeneous disorder, mutations in Gap junction beta 2 [GJB2] gene encoding Connexin 26 [Cx26] protein are the major cause of autosomal recessive and sporadic non-syndromic deafness in many populations. In this study, we have investigated the prevalence of the GJB2 gene mutations using nested PCR pre screening strategy and direct sequencing method. Two hundred and seventy two hearing impaired subjects were studied from 210 families obtained from two large cities of Iran [Tehran and Tabriz]. Twenty four different genetic variants were identified. Cx26 mutations were found in 53 of the 210 families [25.2%] including T8M, 35delG, W24X, R32H, V37I, E47X, 167delT, delE120, Y136X, R143W, R184P, 235delC and V27I+E114G. Homozygosity and compound heterozygosity for the Cx26 mutations were found in 39 of 210 [18.5%] families. Homozygosity for the 35delG mutation was the most common that causes hearing loss in 28 [13.3%] patients. Six novel variants H16R, E101E, K102Q, G200R, 327delG and G130A were detected in this study. As a conclusion, the present survey revealed that the rate of mutation in Cx26 gene in our area is lower than in Europe; nevertheless, this rate is regarded as a considerable cause of deafness in the cited provinces in Iran

Report of a new mutation and frequency of connexin 26 gene [GJB2] mutations in patients from three provinces of Iran

Autosomal recessive and sporadic non-syndromic hearing loss [ARSNSHL] is the major form of hereditary deafness.Mutations in the GJB2 gene encoding the gap-junction protein Connexin 26 have been identified to be highly associated with ARSNSHL. In this study we have analyzed 196 deaf subjects from 179 families having one or more deaf children in 3 proviences of Iran, including Kordestan, Khuzestan and Golestan. The nested PCR prescreening strategy and direct sequencing technique were used to detect the mutations in coding exon of the gene. Altogether 3 GJB2 recessive mutations including 35delG, 167delT and V27I+E114G, were identified in 23 of 179 families [12.8%]. Fourteen of 179 families were observed to have GJB2 mutation in both alleles [7.8%]. A novel variant [R159H] also was found in a deaf family from Khuzestan. Four polymorphisms V27I, E114G, S86T and V153 I also were detected in 7 families. A polymorphism [S86T] was seen in the whole population studied. Our data indicated that the rate of connexin 26 mutations is different in this three Irainian population and is lower than the high frequency of 35delG [26%] reported from Gilan province in the north of Iran

Frequency of a very rare 35delG mutation in two ethnic groups of iranian population

The 35delG mutation in the Connexin 26 gene [Cx26], at the DNFB1 locus is the most common mutation in the patients with autosomal recessive non-syndromic hearing loss [ARNSHL]. We have studied a total of 224 deaf cases from 189 families in two populations of Iran [Sistan va Bluchestan and Hormozgan provinces] by prescreening nested PCR, polyacrylamide gel electrophoresis and consequent direct sequencing method for all cases. The aim of the present work was to find prevalence of GJB2 mutations in the populations studied. Four different GJB2 mutations including 35delG, W24X, R127H and [V27I + E114 G] were identified in 11 of 189 families [5.8%]. Two polymorphisms [V27I and V153I] also were detected in 14 families. A polymorphism S86T was determined in all cases. Homozygote 35delG mutation was found only in 1 of 189 families [0.5%].The rate of Cx26 mutations found in this study was lower than other Iranian populations. So the cause of deafness in the populations studied remains to be detected in other loci or genes