Graphical abstract

Background: Mutation of the beta-globin gene (HBB) interferes with primary mRNA transcription, leading to beta-thalassemia disease. The IVS1nt1 and IVS1nt5 mutations were reported as two of the most prevalent intronic mutations associated with beta-thalassemia major. These mutations may affect the mRNA structure of the human beta-globin (HBB) gene. However, the mechanism by which variation in HBB alters the mRNA structure remains unclear. The objective of this study was to unveil the secondary and tertiary conformation difference of the mutants compared to the wildtype using in silico analysis. Methods: The sequence of HBB was obtained from Ensemble database and mutated manually at nucleotides 143 (IVS1nt1G>T) and 147 (IVS1nt5G>C). The RNA secondary and tertiary structure were performed by ViennaRNA Web Services and 3dRNA v2.0, respectively. Results and Discussion: The results revealed the unique folding characteristics of each mutations for the secondary and tertiary structures. Based on the structure, unwanted folding occurred in the IVS1nt1G>T and IVS1nt5G>C mRNA structures compared to the wild-type structure. This finding was supported by the results of centroid-based analysis and RNA structure analysis, indicating that the larger loops in IVS1nt1 and IVS1nt5 result in an unstable structure. Our study found that intronic mutations affect the mRNA structure of HBB by altering its folding mechanism.

One of the efforts to control the growth of microorganisms is sterilization. The sterilization process can use ozone gas, which is a triatomic form of the element oxygen. Ozone acts as an oxidizing agent capable of destroying the structure of bacterial cell walls, and their molecules are unstable and easily decomposed into oxygen, so that ozone can be applied in sterilization technology for medical devices. In this work, a sterilization box with an ozone generator has been designed with a MQ-131 ozone sensor as an indicator if there is a leak in the box. The output voltage of the ozone generator is 4 kV with a current of 30 mA. We developed an instrument using an Arduino nano as a microcontroller for reading sensor values and displaying sensor values on an LCD monitor. For testing the sterilization effect of the ozone box, a Staphylococcus aureus bacterial sample was used. For the sterilization object, we designated stethoscopes and thermometers as medical equipment. The optimum time for sterilizing medical devices on the sterilization ozone box was 20 minutes, which can reduce the colony of S. aureus bacteria with an ozone concentration of 4.94 ppm.

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Background: Beta globin gene is responsible for producing beta globin chains that stabilize the structure and function of hemoglobin. This gene expression is controlled by complex interactions of transcriptions factors and its regulatory elements in a specific manner. Disturbed beta globin genes may result in hemoglobinopathies, mainly sickle cell disease and beta thalassemia. It seems interesting that several mutations occurring in intronic region results in severe symptoms to beta thalassemia patients, such an IVS1nt5 G>C. This research aimed to analyze RNA structural alteration effected by intronic mutation of beta thalassemia. Methods: The most prevalent mutation of beta thalassemia in Indonesia was obtained from Ithanet. The RNA secondary structure of IVS1nt5 G>C and beta globin gen (HBB) wildtype were performed by RNAStructure, along with probknot prediction. Results: The result showed that intronic mutation caused conformational change in beta globin secondary structure, either for max expect or base pairing probability approach. The mutant had bigger and more loops that diminished the protein stability. Thus, the structure might undergo dysfunction. Conclusion: The comprehensive structural-functional significance of these findings needs further study.