The level of allergens in dust samples collected from selected schools in Shiraz, Iran and its asthma-risk implications.

BACKGROUND: Both home and school are important places where children are exposed to various indoor allergens. This study aimed to identify the profile of indoor allergens in schools and its impact on asthma development. METHODS: A total of 104 classrooms from 52 schools were selected for dust collection during the fall of 2017. The levels of indoor allergens including dust mite (Der f1, Der p1), cat (Fel d1), cockroach (Bla g1) and mouse (Mus m1) were measured by enzyme linked immunosorbent assay (ELISA). The diagnosis of asthma was made in all students of the selected classes by the allergist. The collected data were analyzed using SPSS version 21.0. RESULTS: Out of 2816 students in the selected classes, 180 students were involved with asthma. Students were mostly exposed to Bla g1 (83.1%), followed by Der f1 (51.5%), Mus m 1 (45.5%), Der p1 (8.9%) and Fel d1 (7.9%) in the dust collected from 101 classrooms. Although levels of all studied allergens in the settled dust of the classrooms were low, there was a relationship between Fel d1 in the classroom dust and development of asthma. CONCLUSION: This study showed considerable levels of cockroach allergens in schools. Exposure to cat allergen in our schools played an important role in asthma development; further school-based investigations require evaluating the role of classroom allergen on asthma development.

A new MEFV gene mutation in an Iranian patient with familial Mediterranean fever.

Familial mediterranean fever (FMF) is an inherited autoinflammatory disorder characterized by recurrent episodes of fever and painful inflammation involving the intra-abdominal organs, the lungs and the joints, which is highly prevalent in specific ethnic groups including the Iranians. We report a 12-year-old boy from Iran, with a clinical history of recurrent fever. Based on the suggestive clinical data, mutational analysis revealed the presence of the novel c.1945C>T heterozygous variant in exon 10, which leads to a leucine to phenylalanine change at position 649 of the protein. The mutation was inherited from the mother. This novel mutation lies in exon 10 of the MEFV gene, which encodes for a domain called B30.2-SPRY, located in the C-terminal region of the pyrin protein and contains the most frequent mutations associated with FMF. The present report expands the spectrum of MEFV gene mutations associated with FMF. The uniqueness of this study, compared with other published case reports, consists in the new mutation found in the MEFV gene. In fact, new mutations in this gene are of high interest, in order to better understand the role of this gene in autoinflammation.

Animal allergen sensitization in veterinarians and laboratory animal workers.

BACKGROUND: Animals secrete allergens into the environment and exposure to these in the workplace may cause sensitization. AIMS: To identify the frequency of animal allergen sensitization and symptoms in animal workers. METHODS: Using skin prick tests (SPT), we assessed sensitization to 15 mammal and bird allergens in animal workers and controls. We also recorded symptoms and pulmonary function tests. RESULTS: There were 100 animal workers and 50 controls included in the study. Thirty-six per cent of animal workers and 10% of controls had positive SPT (P < 0.001, odds ratio [OR] = 5.1, 95% CI 1.7-16.0). The most common sensitizations were to horse (16% in animal workers versus 0% in controls, P < 0.01), canary (16% in animal workers versus 2% in controls, P < 0.05, OR = 9.3, 95% CI 1.2-194), cattle (13% in animal workers versus 0% in controls, P < 0.05), cat (12% in animal workers versus 6% in controls, not significant), rabbit and hamster (10% each in animal workers versus 0% in controls, P < 0.05). Allergy symptoms were reported by 52 animal workers, but only 36 of them had positive SPT. Twelve animal workers had abnormal pulmonary function tests and six had positive SPT. CONCLUSIONS: Animal workers are at high risk of occupational sensitization to animal allergens. Exposure should be minimized through control measures and worker education about the risks of exposure and sensitization.

Genetic connections among Turkic-speaking Iranian ethnic groups based on HLA class II gene diversity.

Iran is a linguistically heterogeneous nation where Persian, Turkic and Arabic are the three main language families spoken. Based on their linguistic properties, Qashqais, Turkmens and Azeris are Turkic-speaking people. The purpose of this study was to investigate whether any genetic relationship exists among the Turkic-speaking Iranian subpopulations based on HLA class II gene diversity. HLA-DRB1, DQA1 and DQB1 alleles were identified by PCR-based methods in 100 Qashqais and 66 Turkmens, and the results were compared with our previously published HLA data for Azeris. Despite a number of allelic and haplotypic similarities, Qashqais, Turkmens and Azeris were not in the same clade of the phylogenetic tree. However, based on the results of principal coordinates analysis, they are grouped together with Kurds and Bakhtiaris. Contrary to their common linguistic features, the Turkic-speaking people of Iran are closer to other Iranian subpopulations than to the people of Turkey and central Asia. Overall, it seems that linguistic criteria alone are not able to determine the relationships among these populations, and a combination of different kinds of anthropological information should be used to determine their actual phylogenetic relationships.

Discordant patterns of mtDNA and ethno-linguistic variation in 14 Iranian Ethnic groups.

BACKGROUND/AIMS: Present-day Iran has long represented a natural hub for the expansion of human genes and cultures. That being so, the overlapping of prehistoric and more recent demographic events interacting at different time scales with geographical and cultural barriers has yielded a tangled patchwork of anthropological types within this narrow area. This study aims to comprehensively evaluate this ethnic mosaic by depicting a fine-grained picture of the Iranian mitochondrial landscape. METHODS: mtDNA variability at both HVS-I and coding regions was surveyed in 718 unrelated individuals belonging to 14 Iranian ethnic groups characterized by different languages, religions and patterns of subsistence. RESULTS: A discordant pattern of high ethno-linguistic and low mtDNA heterogeneity was observed for the whole examined Iranian sample. Geographical factors and cultural/linguistic differences actually represented barriers to matrilineal gene flow only for the Baloch, Lur from Yasouj, Zoroastrian and Jewish groups, for which unusual reduced levels of mtDNA variability and high inter-population distances were found. CONCLUSION: Deep rooting genealogies and endogamy in a few of the examined ethnic groups might have preserved ancestral lineages that can be representative of Proto-Indo-Iranian or prehistoric mitochondrial profiles which survived relatively recent external contributions to the Iranian gene pool.

An evolutionary approach to the medical implications of the tumor necrosis factor receptor superfamily member 13B (TNFRSF13B) gene.

Coding variants in tumor necrosis factor receptor superfamily member 13B (TNFRSF13B) have been implicated in common variable immunodeficiency (CVID), but the functional effects of such mutations in relation to the development of the disease have not been entirely established. To examine the potential contribution of TNFRSF13B variants to CVID, we have applied an evolutionary approach by sequencing its coding region in 451 individuals belonging to 26 worldwide populations, in addition to controls, patients with CVID and selective IgA deficiency (IgAD) from Italy. The low level of geographical structure for the observed genetic diversity and the several neutrality tests performed confirm the absence of recent population-specific selective pressures, suggesting that TNFRSF13B may be involved also in innate immune functions, rather than in adaptive immunity only. A slight excess of rare derived alleles was found in patients with CVID, and thus some of these variants may contribute to the disease, implying that CVID probably fits the rare variants rather than the common disease/common variant paradigm. This also confirms the previous suggestion that TNFRSF13B defects alone do not cause CVID and that such an extremely heterogeneous immunodeficiency might be more likely related to additional, still unknown environmental and genetic factors.

The frequency of transforming growth factor-beta1 gene polymorphisms in a normal southern Iranian population.

Several single nucleotide polymorphisms (SNPs) of the transforming growth factor-beta1 gene (TGFB1) have been reported. Determination of TGFB1 SNPs allele frequencies in different ethnic groups is useful for both population genetic analyses and association studies with immunological diseases. In this study, five SNPs of TGFB1 were determined in 325 individuals from a normal southern Iranian population using polymerase chain reaction-restriction fragment length polymorphism method. This population was in Hardy-Weinberg equilibrium for these SNPs. Of the 12 constructed haplotypes, GTCGC and GCTGC were the most frequent in the normal southern Iranian population. Comparison of genotype and allele frequencies of TGFB SNPs between Iranian and other populations (meta-analysis) showed significant differences, and in this case the southern Iranian population seems genetically similar to Caucasoid populations. However, neighbour-joining tree using Nei's genetic distances based on TGF-beta1 allele frequencies showed that southern Iranians are genetically far from people from the USA, Germany, UK, Denmark and the Czech Republic. In conclusion, this is the first report of the distribution of TGFB1 SNPs in an Iranian population and the results of this investigation may provide useful information for both population genetic and disease studies.

Human leukocyte antigen class II allele frequencies and haplotype association in Iranian normal population.

The polymorphism of the HLA class II genes DRB1, DQA1, and DQB1 was investigated in 100 unrelated Iranian individuals from Fars province in Southern Iran, using the restriction fragment length polymorphism (RFLP) method. Subtyping of DRB1*04, *15, and *16 alleles was performed using PCR amplification with sequence specific primes (PCR-SSP). The allele and the haplotype frequencies were calculated. The most common DRB1 alleles were DRB1*11, DRB1*15, and DRB1*04 with a frequency of 25.0%, 14.5%, and 10.5%, respectively. In contrast, the allelic frequency of DRB1*12 and DRB1*08 was very low (1.5% for each). In the DR15 group DRB1*1501 was the most prevalent variant (6.0%). Concerning DR4, the most common alleles were DRB1*0405 and DRB1*0402 (3.5% for each). Interestingly, DRB1*0402 was associated with DQB1*0302 and DRB1*0405 was associated with DQB1*0302 and DQB1*02, the latter being a rare DRB1/DQB1 haplotype in Caucasian individuals. The most frequent DQB1 alleles were DQB1*0301 (31.0%), and DQB1*05 (22.0%). The most frequent DQA1 variants were DQA1*0501 (39.0%) and DQA1*0102 (14.5%). The most common haplotype was DRB1*11-DQB1*0301-DQA1*0501 (25.0%) followed by DRB1*0301-DQB1*02-DQA1*0501 (10%) and DRB1*0701- DQB1*02-DQA1*0201 (6.5%). Data presented in this study suggest that the Iranian population shares some HLA components with populations resident in eastern and southern European countries.

Germline BRCA1 mutations in Iranian women with breast cancer.

BRCA1 is known as a familial early onset breast cancer gene located in the long arm of chromosome 17. Alterations in this gene have been reported in different populations, some of which are population-specific mutations showing a founder effect. In this study, which is the first report on the genetic testing of Iranian women, exon-2 of the BRCA1 gene was sequenced in 80 Iranian breast cancer patients and none of the patients showed the 185del AG mutation or any changes in the sequences of this exon. Among this population, 22 patients having one or more of any kind of tumor in their first-degree relatives were selected and their entire BRCA1 gene coding region was sequenced by direct PCR-sequencing. A novel point mutation in exon-16, with unknown significance, was observed among the breast cancer patients and control subjects. This A/G mutation caused the substitution of Glutamine 1612 with Glycine, with an allele frequency of 38.6 and 52.8% in patients and controls, respectively. In addition, a point mutation in exon-15 and eight other polymorphic alterations were detected which have been reported previously. Three of these polymorphic sites were placed in the intronic part of the gene. To understand the significance of the contribution of the BRCA1 gene in the breast cancer among Iranian, further investigations are needed.

p53 gene alteration and protein expression in Iranian women with infiltrative ductal breast carcinoma.

Genetic alterations in the p53 tumor suppressor gene with or without protein overexpression have been reported to be associated with sporadic breast cancer. To assess the role of p53 in infiltrative ductal breast carcinoma among Iranian patients, p53 protein expression and p53 gene mutations were studied by immunohistochemical analysis and single-strand conformation polymorphism, respectively. The p53 protein was expressed in 25 out of 51 (49%) tumors and p53 gene mutations were detected in 17 out of 37 (46%) tested tumors. No significant correlation was observed between p53 gene mutations and p53 protein expression. There was no significant correlation between p53 abnormalities (mutation and expression) and tumor size, histological grade, nodal status, and progesterone receptor expression. However, a non-statistically significant trend of association (P=0.07) was observed between p53 gene mutations and lack of estrogen receptor. The high percentage of alterations both in p53 gene and protein among southern Iranian breast cancer patients suggests that p53 is probably one of the genes involved in sporadic breast cancer in this area.

Exon-1 polymorphism of ctla-4 gene in Iranian patients with Graves' disease.

Polymorphisms in ctla-4 gene have been shown to be associated with the Graves' disease (GD) susceptibility in different populations in the world. This study was undertaken to disclose the probable association of exon-1 polymorphism of ctla-4 with GD in Iranian patients. A49G polymorphism was investigated in 90 patients and 90 age/sex matched normal healthy controls, using PCR-SSCP and PCR-RFLP methods. Frequencies of AA, AG and GG genotypes among patients were found to be 21 (23.3%), 49 (54.5%) and 20 (22.2%) while these frequencies among healthy controls were 30 (33.3%), 53 (58.9%) and 7(7.8%), respectively. A significant increase of GG genotype and G allele was observed in patients (p = 0.012 and p = 0.025). In conclusion, consistent with the results of most other studies, the presence of a G allele in position 49 of ctla-4 exon-1 is associated with susceptibility to GD in Iranian population.

HLA-DBR 1 alleles and the susceptibility of Iranian patients with breast cancer.

Breast cancer is considered a major malignancy among women worldwide. The contribution of genetic elements to the onset of familial breast cancer has already been established. The current study investigate the alfele frequency of HLA-DRB 1 in 36 primary operable female breast cancer patients from southern Iran by polymerase chain reaction using sequence specific primers (PCR-SSP). Results were compared with those of 36 female control subjects. Statistical analysis was performed and P values were determined for each character. Our results indicated that the frequency of HLA-DRB 1*12 allele is significantly higher in the patient group (p<0.03) compared to the control group. In addition, HLA-DRB1*11 appeared to be as the most frequent allele in the control group (29.2%) and had approximately the same distribution among the patient group (22.5%).

High risk HPV types in southern Iranian patients with cervical cancer.

This study was undertaken to assess the rate of HPV infection in cervical carcinoma among southern Iranian patients. 101 archival cervical carcinoma tissue samples of a 10 year period were studied for the presence of HPV DNA in southern Iran by a polymerase chain reaction method. In addition, the presence of high risk HPV-16 and HPV-18 genotypes was investigated. In total, 88 (87.1%) of the samples were HPV DNA positive, of which 83 were squamous cell carcinomas and 5 were adenocarcinomas. HPV-16 genotype was detected in 26.7% of HPV positive cervical carcinomas; however, none of the samples were positive for the existence of HPV-18 genotype. Collectively, these results suggest that HPV-16 and HPV-18 are not the frequent high risk HPV types in our patients and circulating HPV types in southern Iranian population are different from many other populations.

HLA class II similarities in Iranian Kurds and Azeris.

The genetic relationship between Kurds and Azeris of Iran was investigated based on human leukocyte antigen (HLA) class II profiles. HLA typing was performed using polymerase chain reaction/restriction fragment-length polymorphism (PCR/RFLP) and PCR/sequence-specific primer (PCR/SSP) methods in 100 Kurds and 100 Azeris. DRB1*1103/04, DQA1*0501 and DQB1*0301 were the most common alleles and DRB1*1103/04-DQA1*0501-DQB1*0301 was the most frequent haplotype in both populations. No significant difference was observed in HLA class II allele distribution between these populations except for DQB1*0503 which showed a higher frequency in Kurds. Neighbor-joining tree based on Nei's genetic distances and correspondence analysis according to DRB1, DQA1 and DQB1 allele frequencies showed a strong genetic tie between Kurds and Azeris of Iran. The results of amova revealed no significant difference between these populations and other major ethnic groups of Iran. No close genetic relationship was observed between Azeris of Iran and the people of Turkey or Central Asians. According to the current results, present-day Kurds and Azeris of Iran seem to belong to a common genetic pool.

HLA class II gene polymorphism in Parsees and Zoroastrians of Iran.

Extensive polymorphism of the HLA genes in different ethnic groups has been used as an invaluable tool for anthropological studies. In this study, HLA-DRB1, DQA1 and DQB1 allele frequencies and haplotypes were determined in 72 Parsees and 65 Zoroastrians living in Iran. The predominant DRB1 allele was *1103 = 4 in Parsees and *0701 in Zoroastrians. DQA1*0501 was the most common alleles in both spopulations. The most frequent DQB1 allele was *0301 in Parsees and *0201 in Zoroastrians. DRB1*1103 = 4-DQA1*0501-DQB1*0301 and DRB1*0701-DQA1*0201-DQB1*0201 were the most prevalent haplotypes in Parsees and Zoroastrians, respectively. Significant deviation from Hardy-Weinberg equilibrium was observed in DQA1 and DQB1 loci of Zoroastrians. The former locus also departed from neutrality due to balancing selection. All pairs of the studied loci in this study showed significant linkage disequilibrium. Analysis of molecular variance indicated that the main variation was confined to individuals within the studied populations. Neighbour-joining tree based on Nei's genetic distances according to DRB1 and DQB1 allele frequencies showed that Parsees and Zoroastrians of Iran were located in the same cluster of the phylogenetic tree. Furthermore, Zoroastrians of Iran and Pakistan are very close to each other. This study will serve as a reference for further anthropological studies when the HLA profile of all ethnic groups of Iran is investigated.

Association of IL-18 promoter polymorphisms with predisposition to Type 1 diabetes.

AIMS: IL-18, a potent IFN-gamma-inducing cytokine, is capable of polarizing the immune response to a Th1 phenotype. Recent studies have demonstrated an association between single-nucleotide polymorphisms located at positions -607 (A/C) and -137 (C/G) in the promoter region of IL-18 gene and Type 1 diabetes. The aim of the present study was to determine whether the same polymorphisms of the gene were associated with Type 1 diabetes in Iranians. METHODS: In 112 patients with Type 1 diabetes and 194 non-diabetic control subjects, these two single-nucleotide polymorphisms were analysed by sequence-specific PCR. RESULTS: Allele and genotype frequencies of the IL-18 gene polymorphisms were similar in the whole group of Type 1 diabetic patients and controls. However, categorizing patients according to age at onset of diabetes revealed a significant difference in distribution of the genotypes at position -137 between patients with older age at onset (> 15 years) (GG 49%, GC 34%, CC 17%) and control subjects (GG 57.7%, GC 36.6%, CC 5.7%) (P = 0.027). Frequency of the C allele at position -137 was significantly higher in these patients than in controls (P = 0.038). Moreover, there was an association between -607AA/-137CC genotype combination and susceptibility to Type 1 diabetes in this subgroup of patients (pc = 0.027). CONCLUSIONS: The results of this study show that polymorphisms of IL-18 promoter confer susceptibility to Type 1 diabetes in Iranian individuals with onset at older ages. Further investigations are necessary to clarify the effect of IL-18 variants on immune regulation.

HLA class II (DRB1, DQA1 and DQB1) associated genetic susceptibility in Iranian multiple sclerosis (MS) patients.

The association of HLA class II alleles with multiple sclerosis (MS) has been amply documented. In the present study, the role of HLA class II (DRB1, DQA1 and DQB1) alleles and haplotypes was investigated in 43 unrelated Iranian chronic progressive multiple sclerosis (CP-MS) patients compared with 100 healthy individuals. HLA typing for DRB1, DQA1 and DQB1 was performed by restriction fragment length polymorphism (RFLP). Subtypes of DR4, DR15 and DR16 were defined using polymerase chain reaction (PCR) amplification with sequence-specific primers (PCR-SSP). The results show that, among DR2-positive MS patients and the control group, a positive association with the DRB1*1503, DQA1*0102, DQB1*0602 haplotype (21% vs. 2.7%, P = 0.057, RR = 9.8) and a negative association with the most frequent DR15 haplotype in the control group, DRB1*15021, DQA1*0103, DQB1*0601 (7% vs. 24.3%, P = 0.001), were observed. No significant association was found with the analysed HLA-DRB1, DQA1 and DQB1 alleles.

Molecular analysis of HLA allele frequencies and haplotypes in Baloch of Iran compared with related populations of Pakistan.

The extreme polymorphism in different loci of the human leukocyte antigen (HLA) system has been used as an invaluable tool for anthropological studies. Determination of HLA allele and haplotype frequencies in different ethnic groups is useful for population genetic analyses and the study of genetic relationships among them. In the present study, molecular analysis of HLA-A, -B, -C, -DQA1, -DQB1, and -DRB1 genes has been used to assign HLA allele and haplotype frequencies in 100 unrelated healthy individuals from the Baloch ethnic group of Iran. The results were compared with Baloch and other ethnic groups in the neighboring Pakistan. The results of this study showed that the most frequent HLA class I alleles were A*02011 (20.2%), B*4006 (11.1%), and C*04011 (28.6%). The most common HLA class II alleles were DQA1*0101/2 (42.5%), DQB1*0201 (32%), and DRB1*0301 (29%). Three-locus haplotype analysis revealed that A*11011-B*4006-C*15021 (5.8%) and DQA1*0501-DQB1*0201-DRB1*0301 (22.1%) were the most common HLA class I and II haplotypes, respectively, in this population. Neighbor-joining tree based on DA genetic distances and correspondence analysis according to HLA-A, -B, -DQB1, and -DRB1 allele frequencies showed that Baloch of Iran are genetically very close to Baloch and Brahui of Pakistan. This may reflect an admixture of Brahui and Baloch ethnic groups of Pakistan in the Balochistan province of Iran.

Mutation analysis in F9 gene of 17 families with haemophilia B from Iran.

Seventeen haemophilia B families from Iran were investigated to determine the causative mutation. All the essential regions of the F9 gene were initially screened by conformational sensitive gel electrophoresis and exons with band shift were sequenced. Seven of the 15 mutations identified in these families were novel mutations. The mutations were authenticated in nine families as other affected members or heterozygous female carriers were available for verification.