A novel missense mutation (c.1006C>T) of SPG20 gene associated with Troyer syndrome

The number of gene mutations involved in the hereditary spastic paraplegias is rapidly growing due to the expansion of the frontiers of genomic research by next-generation DNA sequencing platforms. Nevertheless, a comprehensive genetic diagnosis method remains yet unavailable for these diseases. In the current research, an 8-year-old boy with short stature and developmental delay impairment, from a nonconsanguineous family, was referred to our genetic lab. Firstly, based on the physician recommendation, the patient was evaluated by tandem mass spectrometry (MS/MS) for the quantitative examination of amino acids, and then the patient was genetically investigated by karyotype analysis and whole-exome sequencing (WES) technique. Subsequently, targeted Sanger sequencing was applied to confirm the presence of the candidate variant in all the members of the family and screening the other patients for Troyer syndrome. Analysis of inherited metabolic disorders by tandem MS/MS showed the state of all the family members as normal and also karyotyping indicated no chromosomal aberration in the patient. Further investigation by WES technique indicated a homozygous missense variant in the SPG20 gene, c.1006C[T. Targeted sequencing result of the mutation confirmed homozygote state for the affected case and a heterozygote genotype for his parents. The mutation was classified as pathogenic. Detection of novel variants especially pathogenic variant in the SPG20 gene was associated with Troyer syndrome, which encodes a multifunctional protein termed Spartin, assist in improving genotype–phenotype correlation of genetic variants and may facilitate initial diagnosis of Troyer syndrome

[Carrier frequency of alpha thalassemia mutations among newborns in northern Iran]

Background and Objective: Alpha Thalassemia is one of the most prevalent hemaglobinophaties worldwide. Alpha thalasseima patients may represent wide spectrum of symptoms ranging from asymptomatic to severe life threatening anemia. This study was done to assess the carrier frequency of alpha globin gene mutations among newborns in north of Iran

Methods: In this descriptive study, 412 cord blood samples of neonate from Amir Mazandari hospitali were randomly selected during 2012. Genomic DNA was extracted using phenol-chloroform method. Multiplex Gap- PCR and PCR-RFLP methods were applied in order to detect three common gene deletions, one triplication and one point mutation

Results: Total allelic frequency of investigated mutations was 0.0825. The -alpha3.7 deletion with allelic frequency of 0.0485 was the most prevalent mutation among 412 neonates. Allelic frequencies of -alpha4.2, alphaalphaalphaanti3.7 triplication and alpha-5nt mutations were 0.0206, 0.0109 and 0.0024; respectively and -Med double gene deletion was not detected

Conclusion: Most mutated cases had single gene deletion that is asymptomatic while -Med double gene deletion was not detected among the neonates. Therefore, there is low probability of a child birth with Hb H disorder in the region

[Detection of common mutations causing drug resistance to mycobacterium tuberculosis strains among patients with tuberculosis using line probe assay method]

Drug resistance to tuberculosis and especially multiple drug resistance tuberculosis [MDR-TB] variants are a serious problem in tuberculosis patients and make difficulties in controlling the disease. This study was coducted for detection of common mutations causing drug resistance of mycobacterium tuberculosis strains among tuberculosis patients using line probe assay method. In this descriptive study, fifty four sputum samples of tuberculosis patients were randomly selected in health centers of Mazandaran, northern Iran during 2012. After culturing of sputum samples on Lowenstein-Jensen medium, genomic DNA was extracted from colonies using CTAB method. Molecular analysis of mutations causing resistance to five different antibiotics including Isiniazide, Rifampin, Sterptomycine, Amicasin / Canamycine, Kinolon were performed using long probe assay [LPA] method. Out of 54 sputum samples, three [5.5%], three [5.5%], four [7.4%] were resistance to Kinolon, Amicasin / Canamycine and Sterptomycine, respectively. Mutation in KATG was seen in 2 samples resistant to Isiniazide. Mutation in rpoB 516 was seen in 3 samples resistant to Rifampin. Four samples [7.4%] were resistant to the two anti-tuberculosis antibiotics, while three samples were resistant to Sterptomycine and Kinolon and one sample was resistant to Rifampin and Canamycine. 7.4% of sputum samples were resistant to the two anti- tuberculosis antibiotics. Line probe assay is a rapid and suitable method for detecting tuberculosis drug resistance

Beta globin gene haplotypes associated with hemoglobin D-Punjab in northern Iran

Background and Objective: Hemoglobin D-Punjab is one of the variant of hemoglobin caused by a mutation on position 121 of beta globin gene which is frequent in India, PakistanandIran. Heterozygote form of this variant is mainly asymptomatic while in combination with hemoglobin S, severe form of anemia occure. This study was carried out to determine the beta globin gene haplotypes associated with hemoglobin D-Punjab in Northern Iran. Methods: This descriptive study was carried out on families of 18 individuals whom were carriers of hemoglobin D-Punjab in Sari in Northern Iran. Genomic DNA was extracted from peripheral blood samples using Phenol-chloroform st and ard protocol. In order to identify different haplotypes associated with hemoglobin D-Punjab, PCR-RFLP method and family linkage analysis were used. Results: In 17 subjects hemoglobin D-Punjab was linked to [+ - - - - + +] haplotype and in one case association with [- + + - + + +] haplotype was observed. Conclusion: The hemoglobin D-Punjab alleles have mainly unicentric origin and [- + + - + + +] rare haplotype may have different genetic origin or is created as a result of gene recombination

[Prevalence of alloimmunization in major beta thalassemia in northern Iran [2010]]

Repeated blood transfusion is the major treatment for patients with major thalassemia. However due to antigen encounters, it may initiate body reactions, including alloantibodies against red blood cell antigens. This study was done to determine the Prevalence of alloimmunization in major beta thalassemia patients in northern Iran. This descriptive - analytic study was carried out on 218 thalassemic patients [100 males and 118 females] with average age of 22.5 +/- 7 years in northern Iran during 2010. Each sample was tested for the presence of Alloantibodies including C, Cw, Lea, E, Lua, Leb, K, Jkb, N, P1, D, Jka, M, S, Xga, e, Fya, s, c, Fyb, k, Kpa, Jsb, Lub and Coa. Eighty eight cases [40.4%; 95% CI: 33.9-46.9] were positive for the presence of alloantibodies. Alloantibodies against C, Cw, Lea red blood cell surface antigens were the most prevalent [40%]. No significant correlation was found between emergence of alloantibody with the age of initial, frequency and duration of blood transfusion. Alloimmunization is a common observation in thalassemic patients and should be prevented by transfusing compatible blood

[Molecular evaluation of hemoglobin D mutations in Mazandaran province, Iran]

Hemoglobinopathies are among the most prevalent genetic disorders worldwide, and occur as a result of mutations in the gene involved in synthesizing hemoglobin chains. By now more than 1000 defects in hemoglobin chains are discovered. Hemoglobin D [Hb D] is one of these disorders, identified by a single nucleotide mutation on codon 121 of beta globin chain. This study was carried out to evaluate Hb D mutations through molecular methods in Mazandaran province of Iran. This descriptive laboratory study was done on 70 patients with an electrophoresis band in hemoglobin-S zone in Mazandaran province of Iran during 2010-11. Capillary zone electrophoresis was done to find out Hb D in 51 patients. Subsequently, PCR-RFLP was performed to evaluate the samples at molecular level. Molecular investigation revealed all cases are carriers of hemoglobin D-Punjab. Two patients were shown to be homozygote carriers of the abnormal gene. This study showed all Hb D affected patients were carriers of Hb D Punjab

[Comparison of hepatitis C virus genotypes in hemodialysis and nonuremic patients]

Hepatitis C viruses [HCVs], which is an enveloped RNA cense positive, are classified into six major genotypes and multiple subtypes. Infection with this virus has been found to be a major cause of liver disease. Also, HCV infection is quite high among chronic hemodialysis patients. The purpose of the present study was to compare the genotypes of HCV and associated risk factors in hemodialysis patients with positive HCV non uremic patients. Sera sample were taken from population consisted of 113 non uremic patients and uremic ones with HCV who referred to Imam Khomeini nephrology clinic and Sari and Ghaemshahr Dialysis Centers: Case group was consisted of 55 patients with positive HCV hemodialysis disease. The control group consisted of 58 patients suffering from non-uremic positive HCV. Samples were tested with improved Real-Time PCR technique using the appropriate kit. In this study, the mean age of case group was 44.88 +/- 14.6 and for the control group was 46.73 +/- 11.9. Considering the sex of participants, 23 [41.8%] were female patients and 32 [58.2%] were in the case group while 13 female [22.4%] and 45 male [77.6%] were in the control group. The most common genotype of HCV in case group was 1a-b [72.7%] and in control group was 3a [50%]. Significant differences [p<0.05] were seen in HCV genotypes between two case and control groups. BUN and Cratinin in hemodialysis patients showed observable differences in comparison to control group [p<0.05], while PTT, AST, ACT in control group were higher in compare with hemodialysis patients [P < 0.05]. This study showed that the hepatitis C virus genotype and its associated risk factors in hemodialysis patients and non uremic patients is different

role of rheumatoid factor and anti-cyclic citrullinated peptide antibody in diagnosis of rheumatoid arthritis

Rheumatoid Factor [RF] occurs in 70- 90% of patients with established rheumatoid arthritis [RA]. Anti-cyclic Citrullinated peptide antibody [Anti-CCP] detection may be used to confirm the diagnosis of RA in patients with uncharacterized chronic inflammatory arthritis. This study is an attempt to evaluate the role of RF and anti-CCP in the diagnosis and prognosis of rheumatoid arthritis. Blood samples were obtained from 55 patients with established RA and 55 healthy controls. Anti-CCP and RF were measured by ELISA and nephelometry. The patient's demographics, disease duration, DMARDs usage, ESR and radiographic abnormalities were recorded. Fifty five RA patients and 55 healthy age and sex matched controls were enrolled. Odd ratio [OR] was 87.42 [C 1=11.2-680.1] for RF and 27.48 [6/08-124/09] for Anti- CCP. Five patients had used more than 3 DMARDs; all of them were RF positive and 4 were also anti-CCP positive. Eleven patients had erosions in their radiographs; 10 of them were seropositive for RF and 9 positive for anti-CCP. Our study suggests that despite lack of specificity, RF continues to be a central part of the definition of RA due to favorable sensitivity profile and the combined use of RF. Also, anti-CCP is a more powerful diag-nostic and prognostic tool

Evaluation of the correlation between non-alcoholic fatty liver disease and insulin resistance

Non-alcoholic fatty liver disease [NAFLD] is one of the most prevalent liver diseases, being associated with type II diabetes mellitus, obesity and dyslipidemia. It seems that insulin resistance observed in those with type II diabetes or in obese persons plays an important role in the pathogenesis of NAFLD. The aim of this study was to evaluate the relationship between NAFLD and insulin resistance, disregarding obesity and diabetes. This case-control study was performed on those patients with NAFLD referring to Sari Imam Khomeini Hospital in 2007 after obtaining informed consent from patients. After repeated sonography and proved fatty liver, 80 patients were enrolled [34 cases and 46 controls] matched for age, gender and body mass index [BMI]. NAFLD was documented in the case group, using ultrasonographic indices. Those in the control group were healthy. ALT and AST were normal in both groups. Fasting plasma glucose, fasting insulin, 2-hour oral glucose tolerance test [OGTT] and insulin-resistance were compared between the two groups. Insulin resistance was calculated using HOMA-IR formula. There were 20 men [58.8%] and 14 women [41.2%] in the case group with the mean age of 38.0 +/- 9.4 years. Those in the control group were 27 men [58.7%] and 19 women [41.3%] with the mean age of 39.4 +/- 10.5 years. There was no significant difference between the two groups concerning their age, gender or BMI. The mean level of FPG was 91.8 +/- 8.4 mg/dl in the case and 92.5 +/- 10.1 mg/dl in the control group. The mean level of OGTT was 110.9 +/- 18.8 mg/dl in the case group and 103.1 +/- 25.8 mg/dl in the control group and the difference was not significant. The mean levels of fasting insulin were 16.1 +/- 4.2 mg/dl and 10.3 +/- 4.8 mg/dl in the case and control groups, respectively [p<0.001]. Insulin resistance was 3.6 +/- 0.8 micro u/l.mmol/l in the case and 2.3 +/- 1.2 micro u/l.mmol/l in the control group. Since the intervening factors were identical in both groups, we can conclude that the role of insulinresistance in the pathogenesis of NAFLD is not dependent on age, gender and BMI

[Antimicrobial effects of methanolic extracts of some species of stachys and phlomis]

The genus Phlomis and Stachys [Labiatae] are widely distributed in Iran. There are no reports on the antimicrobial activity of some of these plants. The medicinal properties attributed to the genus Phlomis and Stachys prompted us to investigate the antimicrobial activity of the methanolic extracts of Phlomis bruguieri, P. herba-venti, P.olivieri, Stachys byzantina, S. inflate, S. lavandulifolia and S. laxa. The antimicrobial activity of these plants were studied using the disc diffusion methods [10,50, 100, 250, 500, 750, and 100 micro g/disc] and determination of minimum inhibitory concentration [MIC] values [10 micro g/ml to 25 mg/ml] against Staphylococcus aureus PTCC 1112, Streptococcus sanguis PTCC 1440, Escherichia coli PTCC1330, Pseudomonas aeroginosa PTCC 1074, Klebsiella pneumoniae PTCC 1053, Aspergilus niger PTCC 5011 and Candida albicans PTCC 5027. The methanolic extracts of the aerial parts of Phlomis bruguieri, P. herba-venti, P. olivieri, Stachys byzantina, S. inflate, S. lavandulifolia and S. laxa exhibited concentration-dependent antibacterial activity against all tested bacteria. The methanolic extracts were more active against Gram-positive microorganisms [Streptococcus sanguis and Staphylococcus aureus]. The extracts did not show any antifungal activity. The results concluded that the methanolic extracts of these plants have a potential source of antibacterial of natural origin

[Seroepidemiological study of leptospirosis in Ghaemshahr Mazandaran province-Iran, Sept-Oct 2004]

Leptospirosis is a zoonotic spirochetal disease caused by coming into contact with water or soil contaminated with urine of the infected animals such as sheep, hours, cow, rat and etc. Farmers, animal husbands, veterinarians and those practicing aqua sport are at risk. This study was undertaken in 13 villages of Ghaemshahr City [September 2004] in which because of previous epidemiological observation the probability of this disease was high. Study was descriptive cross sectional with randomized case selection. Five ml of venous blood was taken from each case and tested using an ELISA method. Information about age, sex, job, area of living, symptoms such as fever, headache, mialgia, diarrhea, nausea during this or past summer were gathered. Serological data showed that prevalence of leptospirosis was 29.4% .There is no relation between sex and positive serological results [54.8% women and 54.18% men]. Leptospirosis in studied area is epidemic and due to its high morbidity, mortality and economical impact,heath education to people would help to control the disease and chemoprophylaxis can also be used