A Novel Mutation in the OXCT1 Gene Causing Succinyl-CoA:3-Ketoacid CoA Transferase (SCOT) Deficiency Starting with Neurologic Manifestations

  Succinyl-CoA:3-oxoacid CoA-transferase (SCOT) deficiency is an inborn error of ketone body utilization characterized by intermittent ketoacidosis crises. This study reports the first Iranian patient with SCOT deficiency who presented with seizure and hypotonia at birth.Accordingly, she was consequently re-hospitalized due to hypotonia and respiratory distress. Laboratory tests revealed hyperammonemia, ketonuria, and metabolic acidosis. Besides, the plasma glucose level was normal without any other abnormality. Despite treatment with high-dose bicarbonate, severe acidosis persisted. Poor response to treatment raised a significant diagnostic challenge among specialists until genetic investigation identified a homozygous nonsense mutation (c.79G>T; p.Gly27*) in the OXCT1 gene (NM_000436), causing SCOT deficiency. Genetic studies help clinicians achieve a definite diagnosis of such metabolic disorders. In this case, the accurate and early diagnosis of SCOT deficiency opened new therapeutic possibilities, including frequent carbohydrate-rich meals and low fat and protein diet. Moreover, our findings expand the mutational and clinical spectrum of SCOT deficiency

PCSK9 SNP rs11591147 association study with coronary artery disease risk in Iran

Pro-protein convertase subtilisin/kexin type 9 (PCSK9) has been implicated in the regulation of the plasma levels of LDL-cholesterol. The SNP rs11591147 variant in PCSK9 gene is associated with low levels of LDL and reduced risk of coronary artery disease (CAD) in various populations. We therefore, investigated the association and protective role of the SNP rs11591147 with CAD in 94 patients and 94 healthy participants as controls. rs11591147 T>G was genotyped in all subjects by TaqMan Probe Real Time PCR, although we could not observe neither positive or insignificant association between SNP genotypes with CAD incidence. Allele frequencies also remained insignificant after performing comparison analyses between cases and controls. LDL-cholesterol and total cholesterol levels were not associated with the genotypes. Our data indicated that CAD or the plasma level of LDL-cholesterol was not associated with the reduction of CAD incidence in Iranian population, even in a population with high frequency of lipid-connected CAD risk factors. Prospective investigations should include more cases to accurately analyze the effect of SNP rs11591147 in protecting patients from CAD

Melatonin inhibits endothelin-1 and induces endothelial nitric oxide synthase genes expression throughout hepatic ischemia/reperfusion in rats

The production of reactive oxygen species (ROS) and dysfunction of vasculature play a central role in the pathophysiology of hepatic ischemia/reperfusion (I/R) injury. The aim of this study was to evaluate the beneficial effects of melatonin on reducing liver I/R injury in rats. Four study groups were formed: (1) saline - administered, control group (Control), (2) melatonin-administered group (MEL), (3) saline -administered I/R group (I/R) and (4) melatonin-administered I/R group (MEL+ I/R). Melatonin was injected intraperitoneally (15 mg/kg) 20 min before ischemia and immediately after reperfusion. After reperfusion, blood and ischemic liver tissues were collected. The group subjected to ischemia showed a significant increase in the serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels, as well as an increase in hepatic malondialdehyde (MDA) concentration. These increases were significantly inhibited by melatonin. Although, I/R augmented the endothelin-1 (ET-1) gene expression and the level of big endothelin-1 (big ET-1) in liver tissue, melatonin attenuated these increases. Conversely, non-significant decrease in endothelial nitric oxide synthase (eNOS) mRNA expression in I/R group was significantly elevated by melatonin in MEL+ I/R group. Melatonin exerts beneficial effects on ischemia/reperfusion liver injury through its anti-oxidative function as well as regulation of hepatic microcirculation.Key words: Melatonin, oxidative stress, ischemia/reperfusion injury, endothelin and nitric oxide synthase

3-Hydroxyisobutyryl-CoA hydrolase deficiency in an Iranian child with novel HIBCH compound heterozygous mutations

We report a patient presenting with developmental delay, Leigh-like abnormalities on MRI and elevated 3-hydroxyisovaleric acid levels. Upon whole-exome sequencing, he was diagnosed with 3-hydroxyisobutyryl-CoA hydrolase (HIBCH) deficiency, and hence subjected to specific dietary treatment. HIBCH deficiency should be considered in the differential diagnosis of Leigh-like disease and/or organic aciduria. © 2019 The Authors. Clinical Case Reports published by John Wiley & Sons Ltd

The frequency of human leukocyte antigen-DRB1 alleles, using sequence-based genotyping in 68 parents-child trios study in Iranian subjects

Background: The human leukocyte antigen-DRB1 (HLA-DRB1) locus is one of the most polymorphic human loci and has a crucial role in the immune system. Assessing the allelic frequencies of HLA-DRB1 locus would be a fundamental factor in defining the origin of populations, relationships with other populations, disease association studies and the constitution of unrelated bone marrow donor registries. In the current study HLA-DRB1 alleles and their frequencies are determined in a family-based study by DNA sequencing-based typing high-resolution (2 field) level of typing. Materials and Methods: Genomic DNA from 3 members of 68 unrelated families (a total of 204 individuals) was extracted. Exon 2 of DRB1 gene was amplified and performed useing AssignTM SBT v4.7 sequence analysis software.Results: We had DRB1*11:04 with frequency of 0.0931, DRB1*03:01 with 0.0882, DRB1*11:01 with 0.0735, DRB1*13:01 with 0.071 and also alleles DRB1*08:03, DRB1*13:42, DRB1*14:04 and DRB1*14:07 with frequency of 0.0024.Conclusion: A total of 34 different alleles were found in the study subjects with DRB1*11:04, DRB1*03:01, DRB1*11:01 being the most frequent alleles respectively.  

PKD2 mutation in an Iranian autosomal dominant polycystic kidney disease family with misleading linkage analysis data

Background Autosomal dominant polycystic kidney disease (ADPKD) is the most common genetic renal disorder caused by mutation in 2 genes PKD1 and PKD2. Thus far, no mutation is identified in approximately 10 of ADPKD families, which can suggest further locus heterogeneity. Owing to the complexity of direct mutation detection, linkage analysis can initially identify the responsible gene in appropriate affected families. Here, we evaluated an Iranian ADPKD family apparently unlinked to both PKD1 and PKD2 genes. This is one of the pioneer studies in genetic analysis of ADPKD in Iranian population. Methods Linkage reanalysis was performed by regenotyping of flanking microsatellite markers in 8 individuals of the ADPKD family. Direct mutation analysis was performed by Sanger sequencing. Results Mutation analysis revealed a pathogenic mutation (c.1094+1G>A) in the PKD2 gene in the proband. Analyzing 2 healthy and 4 clinically affected members confirmed the correct segregation of the mutation within the family and also ruled out the disease in 1 suspected individual. Misinterpretation of the linkage data was due to the occurrence of 1 crossing over between the PKD2 intragenic and the nearest downstream marker (D4S2929). Homozygosity of upstream markers caused the recombination indistinguishable. Conclusion Although analysis of additive informative polymorphic markers can overcome the misleading haplotype data, it is limited because of the lack of other highly polymorphic microsatellite markers closer to the gene. Direct mutation screening can identify the causative mutation in the apparently unlinked pedigree; moreover, it is the only approach to achieve the confirmed diagnosis in individuals with equivocal imaging results. © 2016. The Korean Society of Nephrology. Published by Elsevier

An efficient trio-based mini-haplotyping method for genetic diagnosis of phenylketonuria

Objective: The phenylalanine hydroxylase (PAH) locus has high linkage disequilibrium. Haplotypes related to this locus may thus be considered sufficiently informative for genetic diagnosis and carrier screening using multi-allelic markers. In this study, we present an efficient method for haplotype analysis of PAH locus using multiplexing dyes. In addition, we explain how to resolve the dye shift challenge in multiplex short tandem repeat (STR) genotyping. Materials and Methods: One hundred family trios were included in this descriptive study. The forward primer of a tetra-nucleotide STR and the reverse primer of a variable number tandem repeat (VNTR) were labeled with three different non-overlapping dyes 5-carboxyfluorescein (FAM), 6-carboxy-N,N,N�,N�-tetramethylrhodamine (HEX) and 6-carboxy-N,N,N�,N�-tetramethylrhodamine (TAMRA). The polymerase chain reaction (PCR) products from each family trio were multiplexed for capillary electrophoresis and results were analyzed using Peak Scanner software. Results: Multiplexing trio products decreased the cost significantly. The TAMRA labeled products had a significant predictable shift (migrated at a slower electrophoretic rate) relative to the HEX and FAM labeled products. Through our methodology we achieve, the less inter-dye shift than intra-dye shift variance. Correcting the dye shift in the labeled products, according to the reference allele size, significantly decreased the inter-dye variability (P<0.001). Conclusion: Multiplexing trio products helps to detect and resolve the dye shift accurately in each family, which otherwise would result in diagnostic error. The dye system of FAM, HEX and TAMRA is more feasible and cheaper than other dye systems

The importance of CDC27 in cancer: molecular pathology and clinical aspects

Background: CDC27 is one of the core components of Anaphase Promoting complex/cyclosome. The main role of this protein is defined at cellular division to control cell cycle transitions. Here we review the molecular aspects that may affect CDC27 regulation from cell cycle and mitosis to cancer pathogenesis and prognosis. Main text: It has been suggested that CDC27 may play either like a tumor suppressor gene or oncogene in different neoplasms. Divergent variations in CDC27 DNA sequence and alterations in transcription of CDC27 have been detected in different solid tumors and hematological malignancies. Elevated CDC27 expression level may increase cell proliferation, invasiveness and metastasis in some malignancies. It has been proposed that CDC27 upregulation may increase stemness in cancer stem cells. On the other hand, downregulation of CDC27 may increase the cancer cell survival, decrease radiosensitivity and increase chemoresistancy. In addition, CDC27 downregulation may stimulate efferocytosis and improve tumor microenvironment. Conclusion: CDC27 dysregulation, either increased or decreased activity, may aggravate neoplasms. CDC27 may be suggested as a prognostic biomarker in different malignancies. © 2021, The Author(s)

Identification of splice defects due to noncanonical splice site or deep‐intronic variants in ABCA4

Pathogenic variants in the ATP-binding cassette transporter A4 (ABCA4) gene cause a continuum of retinal disease phenotypes, including Stargardt disease. Noncanonical splice site (NCSS) and deep-intronic variants constitute a large fraction of disease-causing alleles, defining the functional consequences of which remains a challenge. We aimed to determine the effect on splicing of nine previously reported or unpublished NCSS variants, one near exon splice variant and nine deep-intronic variants in ABCA4, using in vitro splice assays in human embryonic kidney 293T cells. Reverse transcription-polymerase chain reaction and Sanger sequence analysis revealed splicing defects for 12 out of 19 variants. Four deep-intronic variants create pseudoexons or elongate the upstream exon. Furthermore, eight NCSS variants cause a partial deletion or skipping of one or more exons in messenger RNAs. Among the 12 variants, nine lead to premature stop codons and predicted truncated ABCA4 proteins. At least two deep-intronic variants affect splice enhancer and silencer motifs and, therefore, these conserved sequences should be carefully evaluated when predicting the outcome of NCSS and deep-intronic variants

Replication of TCF7L2 rs7903146 association with type 2 diabetes in an Iranian population

The transcription factor 7-like 2 gene (TCF7L2) rs7903146 T allele is constantly associated with Type 2 diabetes in various populations and ethnic groups. Nevertheless, this has not been observed in two studies involving Arab populations. The aim of the present study was to investigate the association between TCF7L2 rs7903146 in an Iranian population. Type 2 diabetes patients (N = 258) and normal healthy control subjects (N = 168) from the same area, were examined. The ARMS- PCR (Amplification Refractory Mutation System) technique, subsequently validated by direct sequencing, was used for genotyping. Allele and genotype frequencies were significantly different between patients and controls TT vs. CT + CC [p 0.0081 OR 3.4 95%CI (1.27-11.9)] and T vs. C allele [p 0.02 OR 1.4 95%CI (1.03-1.9)]. Our data thus confirm the association between the rs7903146 T allele and T2D in an Iranian population, contrary to previous reports in Arab populations. This can possibly be attributed to differences in ethnic background or the effects of environmental factors