Study of rs693 and rs515135 in APOB in people with familial hypercholesterolemia

Objective: APOB-related familial hypercholesterolemia [FH] is the most common hereditary hyperchlosterolemia with an autosomal dominant pattern. A number of APOB variants are the most important risk factors for hyperchlosterolemia. APOB is a large glycoprotein that plays an important role in the metabolism of lipoproteins in the human body. Small changes in the structure and function of APOB can cause major problems in lipid metabolism. Two forms of APOB are produced by an editing process of gene replication. APOB48 is required for the production of chylomicrons in the small intestine and APOB100 is essential in liver for the production of very low density lipoprotein [VLDL] and is also a ligand for LDL receptor [LDLR] that mediates LDL endocytosis

Materials and Methods: In this case-control study, rs693 [in exon 26 of APOB] and rs515135 [5 'end of APOB] single nucleotide polymorphisms [SNPs] were analyzed in 120 cases of familial hypercholesterolemia and 120 controls. Both SNPs were genotyped by polymerase chain reaction-restriction fragment length polymorphism [PCR-RFLP] where PCR products were digested with specific restriction enzymes recognising each single nucleotide polymorphism

Results: This study was analyzed by odds-ratio [OR] and its 95% confidence interval [CI] to examine the association of the two SNPs with familial hypercholostermia susceptibility. Statistical analysis showed that both SNPs were in Hardy- Weinberg equilibrium

Conclusion: We found no significant relationship between rs515135 and familiar hypercholesterolemia. However, there was a significant association between the C allele of rs693 and high familial cholesterol levels. Furthermore, it seems the dominant model of T allele occurrence has a protective role in emergence of disease

Expression and prognostic significance of Bcl-2 and Bax in the progression and clinical outcome of transitional bladder cell carcinoma

To evaluate the mRNA expression ratio of Bcl-2/Bax both in normal and tumoral bladder tissues of patients with transitional cell carcinoma [TCC] of bladder and investigate potential correlation between this expression ratio and clinical outcome. In this experimental study, we used real time-PCR to investigate the expression of Bcl-2 and Bax both in normal and tumoral bladder tissues. The Bcl-2/Bax expression ratio was determined in tumoral bladder tissues of patients with transitional cell carcinoma of the bladder [n=40] and correlation between expression ratios and the emergence of early relapses in a follow-up of 14-30 months was examined. Relapse-free time in 14/31 patients [45.16%] with Bcl-2/Bax>1 was shorter than 9 months [range of 2-9 months] with 5.7 months average median while 17/31 patients [54.84%] with Bcl-2/Bax<1 are currently relapse-free [14-30 months]. Bcl-2 and Bax expression levels were not solely correlated with clinical outcome and progression of carcinogenesis. The mRNA expression ratio of Bcl-2/Bax in tumoral bladder tissues may serve as a significant prognostic indicator in predicting the clinical outcome in low grade non-invasive bladder cancer

[ prevalence of common mutation frequency in K-ras codons 12, 13 in Iranian Colorectal Cancer patients]

Colorectal cancer [CRC] is the most gastrointestinal cancer in United States and Europe. It is the third most common cancer in Iranian men, and fourth in Iranian women. Codons 12 and 13 are the hot spots for mutations in colorectal cancer patients, which encodes the activated RAS protein. According to recent researches, somatic mutations in codons 12, 13 [exon1] of K-ras gene are discovered in 20%-50% human CRCs. The aim of this study was to estimate the contribution of K-ras gene mutations in codons 12, 13 in the incidence, and its association with clinicopathologic information like age, sex, familial history, site of primary and histology in Iranian colorectal cancer patients. In this study, we have analyzed 59 tissue specimens of colorectal cancer patients using PCR/sequencing method for codons 12, 13 of K-ras gene. 20.3% of patients [10 in codon 12 and 2 in codon 13] have shown a point mutation. About 60% of mutations occur in rectum and 41.7% in colon. More than 80% of mutations were in adenocarcinomas and less mutations in mucinous. Most mutations were found over the age of 60. Only two patients [16.6%] had a familial history for cancer. According to low rates of k-ras mutations in codons 12, 13, we can say they are not common in Iranian patients. The mutation pattern for Iranian patients differs from other nationalities. Perhaps we can find point mutations in other exons, and we suggest whole genome sequencing for our patients

[Clinical and molecular investigation of friedreich ataxia in Iranian patients]

Friedreich ataxia [FA] is an autosomal recessive disorder that caused by the expansion of GAA trinucleotide repeat in the first intron of gene X25 [1]. FA is characterized by progressive ataxia and deep tendon areflexia in the lower limbs, dysarthria, skeletal deformities, Cardiomyopathy, muscle weakness and diabetes mellitus may be also found. Cardiomyopathy occurs in almost patients with FA [2]. Cardiomyopathy is the most cause of death in FA patients [3]. Aim of present study was to evaluate the size of GAA repeat and it's correlation with age at onset and cardiomyopathy in these patients. Long PCR testing subsequently confirmed the diagnosing of FA and by identification of GAA repeat, an inverse correlation between size of repeat and age at onset and cardiomyopathy was found

[Mutations in RAG1 and ADA genes in patients with severe combined immunodeficiency [SCID]: report of new mutations]

Severe combined immunodeficiency [SCID] is a rare and mortal disorder with X-linked and autosomal recessive inheritance. Many genes is related to the disease including ADA, RAG1, RAG2, Artemis, CD45, JAK3, IL7R which have different clinical presentation and T and B lymphocytes profile. In this study, we investigated gene mutations in suspected patients referred to the Children Medical Center Hospital, Department of Allergy and Clinical Immunology. Blood tests for patients showed T-B-profile, so we selected the genes that were responsible in T and B cell maturation [ADA, RAG1 and RAG2]. According to our possibilities, we studied ADA and RAG1 genes in patients. We did the test by PCR and Sequencing method. Also total ADA activity [tADA] and its isoenzymes [ADA1 and ADA2] were estimated in patients. Our investigation showed two mutations in ADA gene and three in RAG1 gene. In this study, we offer a new protocol for investigation of RAG1 gene. This is the first study on diagnosis of SCID patients through genetic investigation in Iranian patients

[Epidermolysis bullosa simplex: clinical genetics]

Epidermolysis bullosa [EB] is a group of genetic conditions that cause the skin to be very fragile and to blister easily. Epidermolysis bullosa simplex [EBS] is one of the major forms of the disease. The signs and symptoms of this condition vary widely among affected individuals. Blistering may primarily affect the hands and feet in mild cases. In such cases, blisters usually heal without leaving scars. Severe cases of the disease involve widespread blistering that can lead to infections, dehydration and other medical problem. Researchers have identified four major types of epidermolysis bullosa simplex. They are caused by mutations in the same genes. Four type of EBS includes EBS-WC, EBS-K, EBS-MP and EBS-DM. The mildest form of EBS is EBS-WC. Mutation in the KRT14 and KRT5 genes are responsible for the four major types of EBS. These genes provide instructions for making protein called Keratin 5 and Keratin 14. These tough, fibrous protein works together to provide strength and resiliency to the outer layer of the skin. Mutations in either the KRT14 or KRT5 genes cause cells in the epiderm to become fragile and easily damaged. In this article, we will acquaint with this disease and the clinical diagnosis of each subtype

Enhanced cytochrome C oxidase activity in WBC of COPD patients

Chronic obstructive pulmonary disease [COPD] is characterized by decreased expiratory flow rates, increased pulmonary resistance and hyperinflation. Cytochrome C Oxidase [COX] as a key oxidative enzyme modulates oxygen uptake and catalyzes the oxidation of reduced cytochrome C by molecular oxygen. In vitro studies indicate that the activity of COX can be directly regulated by the presence of molecular oxygen. Thus, a better understanding of the role of COX in patients with COPD can provide an important link between the availability of oxygen to tissues and the regulation of oxygen uptake and energy production in these patients. We studied 42 COPD patients [36 males, 6 females] with clinically stable conditions and 50 [42 males, 8 females] healthy sedentary volunteers of similar age. Whole blood was collected by venipuncture in sodium citrate tubes and WBCs were separated by Ficoll according to standard protocol and lysed with microtube pestle homogenizer. The homogenates were centrifuged and the supernatants were used as a cell extract for COX activity determination. Aliquots of this were assayed for total protein content and COX activity. Analysis of COX activity was performed using COX assay kit. Absolute specific COX activity was normalized for total protein. Relative activities were determined by dividing absolute specific COX activity on absolute specific citrate synthase activity. Mitochondrial COX activity and specific activity [absolute and relative] significantly increased in WBCs of patients with COPD in comparison with control samples [p< 0.05] These results indicated that the activity of COX was increased in WBCs of patients with COPD but whether this is a primary or secondary change relevant to hypoxic condition in these patients is not clear and needs further investigation.

Alteration in antioxidant capacity in patients with chronic obstructive pulmonary disease

Chronic obstructive pulmonary disease [COPD] is a major public health problem that needs greater attention. Variability in the susceptibility to develop COPD is related to both genetic and environmental factors. Oxidative stress and inflammation are the major hallmarks of COPD and antioxidant status can be used as a biomarker to assess the risk of chronic diseases. We used the FRAP [ferric reducing ability of plasma] assay as a simple and powerful test for determination of the total antioxidant capacity of plasma of patients and normal subjects. The patients were selected by cross-sectional method. The mean average age +/- SD of normal subjects and patients was 56 +/- 4 and 60 +/- 2 years respectively. The spectrophotometeric method was used for this assay. The means of the FRAP assays in the patients were higher [about twice] than those of normal subjects. The differences were significant [p < 0.01]. The high levels of antioxidant capacity in the patient group indicated that the antioxidant defense system had been activated due to the oxidative stress and hypoxic condition. A though, FRAP assay can probably be used for demarcation of severity and risk of developing COPD, clinical follow-up and further investigation are required for the assessment of this hypothesis

[Association study between spinocerebellar Ataxia [SCA] disease and mitochondrial damages as a source of ATP producer in these patients]

The spinocerebellar ataxias [SCA] comprise a heterogeneous group of severe late-onset neurodegenerative diseases promoted by the expansion of a tandem-arrayed DNA sequence that modify the primary structure of the protein. SCA is a genetic disease with multiple types, each of which could be considered a disease in its own right, and cannot be differentiated rapidly from each other on a clinical basis. As with other forms of ataxia, SCA results in unsteady and clumsy motion of the body due to a failure of the fine coordination of muscle movements, along with other symptoms. A variety of deadly diseases are attributable to a large number of accumulated mutations in mitochondria. Percentage levels of mutant mtDNA in blood have been reported in some patients with neurological disease, usually in association with much higher levels of mutant mtDNA in skeletal muscle. Mitochondrial DNA [mtDNA] defects may present with cerebellar ataxia. MtDNA rearrangements are usually sporadic and may cause ataxia. MtDNA point mutations may be responsible for the ataxia seen in some SCA mutation negative families. Point mutations such as those affecting tRNALeuUUR are particularly common in human mitochondrial diseases. Genomic DNA of 20 patients with clinical symptoms of SCAs was purified from peripheral blood and screened for deletions in mitochondrial DNA [mtDNA]. Genetic analysis Segments of the genes SCA1, SCA2, MJD [SCA3], CACNA1A [SCA6], and SCA7 harboring the CAG-repeat region were amplified in five separate reactions for molecular diagnosis for each patient. Also the sequencing of tRNA[Leu[UUR]], tRNA[Lys], ATPase 6, ATPase8,COII, COIII, ND1, 16srRNA of mtDNA was performed in patients with genetically affected SCA to find out if they harbor any mutation in these regions or not

[Investigation of interferon-gamma receptor-1 polymorphism in Iranian patients affected with pulmonary tuberculosis]

Tuberculosis is one of the most common infectious diseases in the world. In recent years, genetically approach has been developed. One of the interesting gene for investigator is IFN- gamma R1. In this study we determind susceptibility to tuberculosis with polymorphism of IFN- gamma R1 gene. Fifthly patients with smear positive tuberculosis have been chosen randomly. They were matched with 54 healthy controls with no history of TB. Polymorphism at 395 codon of IFN- gamma R1 gene was detected with Newport method. Mean age of patients and control were 55 13.5 years respectively. Demographic characteristic had no +/- 20 and 53 +/- difference within two groups. One patient in case group had heterozygote mutation at IFN- gamma R1 gene. In control group there were no mutations. Genetically susceptibility to TB was not seen in 395 colon of IFN- gamma R1 in Iranian TB sample and polymorphism of this loci has occur in 2% of TB patients and 0.96% of total study population

Detection of 22q 11.2 microdeletions by Semiquantitaive multiplex PCR [SQMPCR]

22q11.2 chromosomal region is a hot spot for many cytogenetic rearrangements especially micro deletions which are responsible for DiGeorge and VeloCaridoFacial syndromes. The most characteristic sign in these patients is congenital cardiac conotruncal anomalies. The gold standard diagnostic test for these micro deletions is FISH [Fluorescent In Situ Hybridization]. However this diagnostic technique has some drawbacks such a high final cost and low sensitivity in smaller and uncommon micro deletions found in this region. The aim of this study was to introduce a less expensive and a priori more sensitive molecular method to help small and peripheral laboratories to find genetic causes of congenital heart diseases and DiGeorge syndrome. 10 patients with congenital conotruncal anomalies and symptoms of DiGeorge syndrome were included in this study. These patients had been analyzed by FISH probe TUPLE1 before the inclusion. 3 normal persons were included as normal controls for micro deletion region. Semi Quantitative Multiplex PCRs were designed based on known markers in and out of the region of interest. Results were analyzed by Total Lab software. 4 patients showed a decrease in gene dosage more than 60% compared to normal persons. FISH analysis found only one patient with micro deletion. The designed method based on semi quantitative PCR was able to find 4 patients [40%] with micro deletion in a population of 10 patients with congenital cardiac anomalies. This techniques was also able to find micro deletions in three FISH negative patients. Molecular diagnosis of micro deletions is supposed to be more sensitive than FISH in small micro deletions. This study confirms the presence of a typical deletions in Iranian patients and shows that the applied technique can detect some FISH negative patients. However further studies are needed to determine the sensitivity and specificity of the mentioned molecular diagnosis. It seems that this can be used at least for the patients with typical phenotypic features of 22q11Ds and negative FISH results

[Prevalence of MT DNA mutation in type II diabetes mellitus]

Cardiovascular diseases are the first cause of death in Iran and hypercholesterolemia is one of the most important risk factors. This problem could be partially managed by dietary modifications such as supplementation of diet with dairy products and probiotic bacteria. The aim of this study was to comprise the effect of consuming a probiotic yoghurt and ordinary yoghurt on serum cholesterol levels in mild to moderate hypercholesterolemia. This randomized cross-over trial was conducted on 14 healthy subjects aged 40-65 years with total serum cholesterol 200-300 mg/dl. We asked the subjects to avoid consumption of yoghurt for a two-week pre-study period and add 300 g/day of milk to their diet. Then they were randomly allocated to 2 groups to receive either 300 g of ordinary yoghurt [fermented with S.thermophilus and L.bulgaricus] or probiotic yoghurt [fermented with a starter composed of L.acidophilus and B.lactis plus bacteria in ordinary yoghurt] for 6 weeks as substitution for milk. After a wash-out period of 7 weeks, the cross-over was made [those consuming probiotic yoghurt, changed to ordinary yoghurt and vice versa] and the study lasted for another 6 weeks. Anthropometric measurements, a 3-day dietary recall and blood lipid tests were done at the beginning [after a 2-week elimination period of yoghurt from diet] and at the end of each period. Comparison of weights, BMIs and dietary confounding factors during the period of consuming ordinary yoghurt and probiotic yoghurt, did not show any significant differences. Consumption of probiotic yoghurt in comparison with ordinary yogurt caused a significant decrease in total serum cholesterol [P=0.049]. Comparison of other blood lipid criteria did not show any significant differences between these two periods. Consumption of the yoghurt containing two probiotic bacteria, L.acidophilus and B.lactis, in comparison with ordinary yoghurt causes reduction in total serum cholesterol in mild to moderate hypercholesterolemia