Alpha globin gene alterations modifying the phenotype of homozygous beta thalassaemia.

The phenotype of ß-thalassemia varies widely. The primary determinant is the type of beta-globin gene mutation; however, there are secondary and tertiary modifiers also as associated alpha mutations, polymorphisms, as well as coinheritance of mutations affecting other related systems. Co-inheritance of alpha thalassemia mutations is known to ameliorate the severity of HbE-ß thalassemia. However, the role of alpha globin gene alterations (deletions and triplication) is not well illustrated in homozygous ß-thalassemia. Here we evaluated the role of alpha globin gene alterations in 122 ß-thalassemia patients having IVS1-5 (G > C) homozygous mutation. ß-thalassemia mutations were detected by ARMS PCR and alpha mutations by GAP-PCR. Gene expression by qRT-PCR. Out of 122 cases, 15 patients had alpha 3.7 triplications (ααα3.7anti), 24 had alpha 3.7 kb deletion (-α3.7) mutation and three patients had 4.2 kb deletion (-α4.2). Patients were divided into two groups, requiring less than 8 units (NTDT) and more than 8 units (TDT) of blood transfusion per year (≥8U BT/year). The percentage of alpha deletion was significantly (p = 0.0042) high in NTDT (42.1%) as compared with TDT (13.2%). Conversely, the proportion of alpha triplication is high in the TDT as compared with NTDT. Even mean serum ferritin level was found to be significantly high in patients having alpha triplication as compared with those having alpha deletions (p = 0.0184) and normal alpha gene (p = 0.0003). α/ß globin ratio was highest in TDT patients with alpha triplication and lowest in NTDT patients with alpha-del. The results show that concurrent inheritance of alpha gene alterations influences the phenotypic severity of homozygous ß-thalassemia.

Seroprevalence of ToRCH Pathogens among Children Admitted to a Tertiary Care Hospital in Eastern India for Cataract Surgery and Cochlear Transplantation.

AIMS: The study aims to investigate the presence of TORCH infections in a child with bilateral cataracts and deafness and report the ToRCH-serology screening profile (Toxoplasma gondii (TOX), rubella (RV), cytomegalovirus (CMV), and herpes simplex virus (HSV-I/II)) in pediatric cataract and deafness. METHODS: Cases that had a clear clinical history of congenital cataracts and congenital deafness were included in the study. The study population consisted of 18 bilateral cataracts and 12 bilateral deafness child who was admitted to AIIMS Bhubaneswar for cataract surgery and cochlear implantation, respectively. Sera of all children were tested qualitatively and quantitatively for IgG/IgM-antibodies against ToRCH agents in a sequential manner. RESULTS: Anti-IgG antibodies against the torch panel were detected in all cataract and deafness patients. Anti-CMV IgG was detected in 17 of 18 bilateral cataract children and 11 of 12 bilateral deaf children. The rates of anti-CMV IgG antibody positivity were significantly higher. In the cataract group, 94.44% and in the deafness group, 91.66% of the patient was Anti-CMV IgG positive. Besides this, 77.7 % of the patient from the cataract group and 75% from the deafness group was anti- RV IgG antibody positive. In bilateral cataract patients, IgG-alone seropositive cases were mostly attributed to CMV (94.44%; 17/18), followed by RV (77.70%; 14/18), HSV-I (27.70%; 5/18), TOX (27.70%; 5/18), and HSV-II (16.60%; 3/18). In bilateral deafness patients, the spectrum of IgG alone seropositive cases was almost the same except for TOX (0/12). CONCLUSION: The current study recommends interpreting ToRCH-screening in pediatric cataracts and deafness with caution. Interpretation should include both serial qualitative and quantitative assays in tandem with clinical correlation to minimize diagnostic errors. The sero-clinical-positivity needs to be tested in older children who might pose a threat to the spread of infection.

Restoration of Altered Oncogenic and Tumor Suppressor microRNA Expression in Breast Cancer and Colorectal Cancer Cell using Epicatechin.

BACKGROUND: MicroRNAs (miRNA) are small non-coding RNAs that regulate the function of mRNA post-transcriptionally in a tissue-specific manner. miRNA expressions are heavily dysregulated in human cancer cells through various mechanisms, including epigenetic changes, karyotype abnormalities, and miRNA biogenesis defects. miRNAs may act as either oncogenes or tumor suppressors under different conditions. Epicatechin is a natural compound found in green tea which possesses antioxidant and antitumor properties. OBJECTIVE: The objective of this study is to investigate the effect of epicatechin treatment on the expression level of several oncogenic and tumor suppressor miRNAs in breast and colorectal cancer cell lines (MCF7 and HT-29) and identify its mechanism of action. METHODS: The MCF-7 and HT29 cells were treated with epicatechin for 24 hours and untreated cells were considered control cultures. miRNA was isolated and qRT-PCR was used to measure the expression profile changes of different oncogenic and tumor suppressor miRNAs. Furthermore, the mRNA expression profile was also screened at different concentrations of epicatechin. RESULTS: Our results showed several-fold changes in miRNAs expression level, which is cell line specific. Also, epicatechin at different concentrations induces biphasic changes in mRNA expression levels in both cell lines. CONCLUSION: Our findings first time demonstrated that epicatechin can reverse the expression of these miRNAs and may trigger the cytostatic effect at a lower concentration.

Phytocompounds curcumin, quercetin, indole-3-carbinol, and resveratrol modulate lactate-pyruvate level along with cytotoxic activity in HeLa cervical cancer cells.

Cancer cells meet their energy need by predominantly increased uptake of glucose, high rate of glycolysis, and increased production of lactate even in the presence of adequate oxygen.  This process was proposed by Otto Warburg and named after him as the Warburg effect. The development of drugs that target glucose intake and aerobic glycolysis or lactic acid secretion of cancer cells is a newer approach for drug discovery. We have tested five purified plants-derived compounds such as curcumin, quercetin, ellagic acid, resveratrol, and indole-3-carbinol in HeLa cells for cytotoxicity, inhibition of metastasis, and modulation of lactate-pyruvate metabolism. Standard biochemical methods were used for glucose, lactic acid, and pyruvic acid measurement. The cell viability was determined by MTT assay. Cell migration was checked by wound healing assay. A dose-dependent cytotoxic effect and inhibition of cell migration were observed in all the tested compounds. A decrease in the lactate and increase in pyruvate level was observed in all the tested compounds except ellagic acid. Our finding suggests that tested phytocompounds are associated with the metabolic reprogramming of cancer cells and execute the cytotoxic effect. These compounds could be used for cancer prevention and therapy.