Pathogenic Homozygous Mutations in the DBT Gene (c.1174A>C) Result in Maple Syrup Urine Disease in a rs12021720 Carrier.

OBJECTIVE: Maple syrup urine disease (MSUD; OMIM #248600) is an autosomal recessive metabolic disorder in the catabolism of branched-chain amino acids (leucine, isoleucine, and valine) and may be lethal if untreated in affected newborns. METHODS: Single-nucleotide polymorphism haplotyping and Sanger sequencing of BCKDHA, BCKDHB, and DBT genes were performed in a cohort of 10 MSUD patients. RESULTS: We identified a 16.6 Mb homozygous region harboring the DBT gene in an Iranian girl presenting with MSUD. Sanger sequencing revealed a pathogenic homozygous variant (NM_001918.3: c.1174A > C) in the DBT gene. We further found a controversial variant (rs12021720: c.1150 A > G) in the DBT gene. This substitution (p.Ser384Gly) is highly debated in literature. Bioinformatics and cosegregation analysis, along with identifying the real pathogenic variants (c.1174 A > C), lead to terminate these various interpretations of c.1150 A > G variant. CONCLUSION: Our study introduced c.1150 A > G as a polymorphic variant, which is informative for variant databases and also helpful in molecular diagnosis.

Evaluation of peripheral blood NK cell subsets and cytokines in unexplained recurrent miscarriage.

BACKGROUND: Recurrent miscarriage is considered as one of the main problems in women's reproductive health. The aim of the present study was to evaluate the natural killer cells (NK cells) and cytokines in unexplained recurrent miscarriage and fertile women. METHODS: In this case-control study, 40 women with unexplained recurrent miscarriage were assigned to the case group and 40 fertile women were assigned to the control group. NK cell subsets (CD56+ CD16+/CD56+ CD16-) and cytokines (IL-2/IL-12) levels in the peripheral blood (PB) were used for assessing immunologic problems. The percentage of peripheral blood NK cells (CD56dim/bright) was identified by flow cytometry. RESULTS: The obtained results showed a significant difference in CD56+ CD16+ and CD56+ CD16- between the two groups. Also, there was no significant difference in the IL-2 and IL-12 between the two groups. A cut-off value of ≥5.25% (p < 0.001) and ≥3.4% (p < 0.015) for the increased percentage of CD56+ CD16+ and CD56+ CD16-cells in the PB become predictive of recurrent miscarriage. CONCLUSION: Increased NK cells in the PB of women with recurrent miscarriage strongly establish prospective researches to recognize the predictive value of these parameters in the evaluation of patients with recurrent miscarriage.

Histological grading and staging of liver and its relation to viral loads in chronic anti-HBe positive hepatitis.

BACKGROUND: Hepatitis activity index (HAI) and fibrosing stage are two important findings during the evaluation of liver samples in patients with chronic hepatitis B. The aim of this study was to determine the HAI and fibrosing stage in patients with anti-HBe positive chronic hepatitis B. METHODS: Liver biopsy slides of 72 patients were evaluated at the Department of Pathology in two teaching hospitals of Babol University of Medical Sciences, Iran, from April 2006 to August 2011. Total HAI or grading as well as its components including piecemeal necrosis, confluent necrosis, spoty necrosis, portal inflammation and fibrosis scores or staging in considering with viral loads more or less than 10(5) copies/ml were enumerated according to Ishak scoring system. RESULTS: The mean age of these patients was 34.4±12 years. Fifty-six patients had viral load> 10(5) copies/ml. Piecemeal necrosis and grading scores with viral load (10(3),10(3)-10(5) and >10(5) copies/ml) were 0.8±0.7, 0.9±0.4, 1.8±1 and 3.8±1.9, 4.4±2, 5.9±2.6, respectively (p=0.005 and p=0.04, respectively). There was not any significant difference with fibrosis stage regarding different viral loads. In total, 18 cases had fibrosis scores > 1 and 24 cases had confluent necrosis. HAI≥4 was seen in 29 (60.4%) of the 48 cases without confluent necrosis and in 23 out of 24 cases with confluent necrosis (p=0.007). CONCLUSION: The results show that piecemeal necrosis and higher grading scores are associated with higher viral loads. The presence of confluent necrosis is associated with more severe diseases.

Evaluation of the need for treatment on 72 subjects with anti-HBe positive chronic hepatitis B.

BACKGROUND: Viral load and alanine aminotransferase (ALT) levels may not be the key points for making a decision in the treatment of anti-HBe positive chronic hepatitis B. The purpose of this study was to assess the histological evaluation of liver to find the need for treatment on 72 patients with anti-HBe positive chronic hepatitis B. METHODS: The liver biopsy slides of the 72 patients (56 subjects with viral load >10(5) with any ALT levels, 16 cases with viral load <10(5) copies/ml with abnormal ALT levels) were evaluated at the Department of Pathology, Babol University of Medical Sciences, Iran from April 2006 to August 2011. Ishak Scoring system was used to determine the hepatitis activity index (HAI) and fibrosis score. Those with total score >3 were considered for treatment. Data were collected and analyzed. RESULTS: The mean age of the patients was 34.4±12 years. The mean ALT level was 105±10.5 IU/L. The mean HAI with viral loads more or less than >10(5) was 5.9±2.6 and 4±1.9, respectively (p=0.04). HAI >3 was seen in 9 (56.3%) and in 43 (76.8%) subjects with viral loads <10(5) and > 10(5) copies/ml (p<0.05). No fibrosis was seen in 25 (34.7%) of these cases. Fibrosis stage regarding viral loads more or less than10(5) copies/ml was equal (p=0.12). The need for treatment was seen in 62 (86%) patients. CONCLUSION: The results show that any viral load values may cause significant injuries that need to treatment. Liver biopsy is indicated in any case of anti-HBe with any viral loads with increased ALT levels.