The Deafness-Associated Mitochondrial DNA Mutation at Position 7445, Which Affects tRNASer(UCN) Precursor Processing, Has Long-Range Effects on NADH Dehydrogenase Subunit ND6 Gene Expression

The pathogenetic mechanism of the deafness-associated mitochondrial DNA (mtDNA) T7445C mutation has been investigated in several lymphoblastoid cell lines from members of a New Zealand pedigree exhibiting the mutation in homoplasmic form and from control individuals. We show here that the mutation flanks the 3' end of the tRNASer(UCN) gene sequence and affects the rate but not the sites of processing of the tRNA precursor. This causes an average reduction of ~70% in the tRNASer(UCN) level and a decrease of ~45% in protein synthesis rate in the cell lines analyzed. The data show a sharp threshold in the capacity of tRNASer(UCN) to support the wild-type protein synthesis rate, which corresponds to ~40% of the control level of this tRNA. Strikingly, a 7445 mutation-associated marked reduction has been observed in the level of the mRNA for the NADH dehydrogenase (complex I) ND6 subunit gene, which is located ~7 kbp upstream and is cotranscribed with the tRNASer(UCN) gene, with strong evidence pointing to a mechanistic link with the tRNA precursor processing defect. Such reduction significantly affects the rate of synthesis of the ND6 subunit and plays a determinant role in the deafness-associated respiratory phenotype of the mutant cell lines. In particular, it accounts for their specific, very significant decrease in glutamate- or malate-dependent O2 consumption. Furthermore, several homoplasmic mtDNA mutations affecting subunits of NADH dehydrogenase may play a synergistic role in the establishment of the respiratory phenotype of the mutant cells

Experimental Investigation of Leakage Flow Rate and Fluidisation beneath Polyethylene Pipes in Non-Uniform Soils

This is the final version. Available on open access from MDPI via the DOI in this recordData Availability Statement: Some or all of the data, models, or code that support the findings of this study are available from the corresponding author upon reasonable request.Soil fluidisation around buried pipes is one of the water leakage effects that has a direct influence on the ultimate failure of pipelines. In this research, using a laboratory model, the fluidisation caused by water leakage from three cracks with three lengths (14, 17, and 20 mm) and a 3 mm diameter hole for five pressures (1.5–5.5 bar) in non-uniform soils has been evaluated. The experiments were carried out both for pipes buried in soil, as well as exposed pipes. In the buried pipe tests, leakage flow rate, fluidisation, and mobile bed zone dimensions were investigated. The results showed that the increase in leakage flow rate due to an increase in pressure and crack length in exposed pipes is higher than in buried pipes. The exponent of the leakage–pressure relationship was obtained between 0.40 and 0.47 for the hole and between 0.8 and 1.9 in the crack. Observing different development patterns for fluidisation and mobile bed zones in cracks and holes, new relationships are presented for the height, width, and cross-sectional area of the leakage zones

Experimental study of head shape effects on shear stress distribution around a single groyne

River hydrodynamicsInteraction with structure

Comparison of Propolis and Calcium Hydroxide in terms of Mineralization and Cytotoxicity Using Dental Pulp Stem Cells

Objectives: This study aimed to compare the in vitro cytotoxic activity of propolis, a bioactive material made by the honeybee, and calcium hydroxide (CH) and their effect on formation of mineralized nodules by human dental pulp stem cells (HDPSCs).Methods: In this in vitro study, HDPSCs were obtained from the Cellular and Molecular Oral Biology Laboratory of School of Dentistry, Shahid Beheshti University of Medical Sciences. In order to evaluate the proliferative effect of propolis and CH, HDPSCs were incubated with different concentrations of propolis (0-32mg/mL) and CH (0-4.8 mg/mL). Twenty-four and 48 hours later, the methylthiazolyl diphenyl-tetrazolium bromide (MTT) assay was carried out to evaluate the proliferation potential and viability of HDPSCs treated with propolis and CH. The effect of propolis and CH on mineralization of HDPSCs was assessed by alizarin red staining.Results: The MTT assay revealed that propolis at its highest concentration caused the greatest proliferation after 24 and 48 hours. Alizarin test showed that the lowest concentrations of CH and propolis at 14 days induced the formation of calcium nodules but at 21 days, propolis was deposited on the cells and calcification was not well recognizable.Conclusion: Propolis led to higher cell vitality at all concentrations in comparison to CH. However, due to its deposition on the cells, its effects on mineralization at 48 hours could not be determined

Association of TNF-α G308A gene polymorphism in essential hypertensive patients without type 2 diabetes mellitus

This study aims to investigate the effects of tumor necrosis factor alpha (TNF-α) G308A gene polymorphism on essential hypertension (EHT) with or without type 2 diabetes mellitus (T2DM). The project was conducted on buccal epithelial and blood cells for case and control patients, respectively. Epithelial cells were obtained from the inner part of the cheeks. Techniques including DNA extraction, polymerase chain reaction (PCR), and restriction fragment length polymorphism (RFLP) were utilized to assess biomarkers of DNA damage. Our results demonstrated significant differences between wild and mutated genotypes among EHT patients without T2DM. We also found a significant association between wild and mutated allele frequencies in EHT patients (P < 0.05). Clinical characteristics between the groups (EHT with or without T2DM and controls) showed statistically significant association (P < 0.05). Overall, we show that G308A polymorphism of the TNF-αgene may be a significant genetic risk factor for EHT without T2DM patients in Malaysia