Background: After ruling out the most common causes of severe hemolytic anemia by routine diagnostic tests, certain patients remain without a diagnosis. The aim of this study was to elucidate the genetic cause of the disease in these patients using next generation sequencing (NGS). Methods: Four unrelated Iranian families including six blood transfusion dependent cases and their parents were referred to us from a specialist center in Tehran. There was no previous history of anemia in the families and the parents had no abnormal hematological presentations. All probands presented severe congenital hemolytic anemia, neonatal jaundice and splenomegaly. Common causes of hemolytic anemia were ruled out prior to this investigation in these patients and they had no diagnosis. Whole exome sequencing (WES) was performed in the probands and the results were confirmed by Sanger sequencing and subsequent family studies. Results: We identified five variants in the PKLR gene, including a novel unpublished frameshift in these families. These variants were predicted as pathogenic according to the ACMG guidelines by Intervar and/or Varsome prediction tools. Subsequent family studies by Sanger sequencing supported the diagnosis of pyruvate kinase deficiency (PKD) in six affected individuals and the carrier status of disease in their parents. Conclusion: These findings show that PKD is among the rare blood disorders that could remain undiagnosed or even ruled out in Iranian population without performing NGS. This could be due to pitfalls in clinical, hematological or biochemical approaches in diagnosing PKD. Furthermore, genotyping PKD patients in Iran could reveal novel mutations in the PKLR gene.
Background: Inflammatory events following brain death (BD) decrease the quality of donor organs which can affect the outcome of the transplantation. It experimentally and clinically is verified that the level of inflammatory cytokines is increased in the BD of the donor results. In experiments, the in vitro use of vitamin C can successfully decrease the levels of inflammatory cytokines. Objectives: The aim of this study was to investigate the influence of vitamin C administration on the inflammatory status of the graft in BD liver donors. Methods: This study was conducted at Nemazee Hospital's transplant center (Shiraz, Iran). In this interventional study, the BD liver donors (n = 40) were randomly divided into two groups. The control group received only the routine intensive care unit (ICU) considerations, and the intervention group was treated with Vitamin C before harvesting organ (100 mg/kg, initially followed by 100 mg/kg/6h until organ removal). Blood samples were taken from BD patients in the intervention and control groups 3 times: 6 hours before operation, immediately after laparotomy, and immediately prior to clamping the aorta to assess the gene expression ratio of Interleukin 1 beta (IL1B) and Interleukin 10 (IL10) cytokines using real time polymerase chain reaction (PCR) (n = 40). Soluble serum cytokines were measured by enzyme-linked immunosorbent assay (ELISA) (n = 24). Results: No significant differences were observed in mRNA expression ratio of IL1B and IL10 between two groups. Despite an acceptable decrease in serum concentration of the inflammatory markers IL1B and IL10 at time point 2 minus time point 1 (T2-1), no significant differences were observed in vitamin C-treated BD donors compared with the control donors. There was a significant difference among aspartate aminotransferase (AST) and alanine aminotransferase (ALT) changes on the 3rd to 1st days after operation between the control and intervention recipients (P < 0.05).
Conclusions:The present study suggests a beneficial effect of vitamin C adsministration on post-transplant function of the liver from BD liver donors.
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