Cloning and expression of choline dehydrogenase gene (betA) of escherichia coli in yeast saccharomyces cerevisiae

The complete DNA sequence of betA, encoding choline dehydrogenase of Escherichia coli has been cloned in a yeast expression-secretion vector and expressed in Saccharomyces cerevisiae DBY 746. The recombinant plasmid (designated as pYYF9) was stably maintained in yeast cells at a level of 24% after 30 generations. Gene expression analysis was carried out at transcription and translation levels. The niRNA analysis by DNA-RNA hybridisation using a specific betA probe did not reveal any signal of transcription of the cloned betA gene in yeast. Protein analysis by SDS-PAGE did not show any specific protein secreted into the yeast supernatant either. However, analysis of the int racellular proteins of yeast demonstrated differences of protein profile between the untransformed with the transformed cells. Keywords : choline dehydmgenase gene (betA) --gene expression â Saccharomyces cerevisia

Tetra‐primer amplification refractory mutation system (ARMS) PCR used to detect 3’UTR rs1948 mutation in CHRNB4

Rs1948 A>G is a single nucleotide variation (SNV) in the 3’‐UTR of CHRNB4. Genotyping the synonymous CHRNB4 rs1948 may be useful in identifying a lung cancer susceptibility gene. The study aimed to develop a simple and easy tetra‐primer amplification refractory mutation system (ARMS PCR) for CHRNB4 rs1948. The following steps were taken to optimize tetra‐primer ARMS PCR: 1) determining the gene sequence and position of a single mutation; 2) developing outer and inner primers; 3) amplification of target gene fragments via PCR using an outer primer; 4) genotyping PCR product using Sanger sequencing; 5) determining the optimal annealing temperature and PCR cycle; 6) determining optimal outer and inner primer ratio; and 7) testing the reproducibility of the PCR program and final validation with Sanger sequencing. Genotype (PCR result) was visualized with 3% agarose gel electrophoresis. Optimum condition was determined as annealing temperature of 64.8 ºC and 35 cycles, outer and inner primer ratio of 1:6, and DNA volume of 3 µL. Sanger sequencing confirmed the results of the tetra‐primer ARMS PCR and it was shown that ARMS PCR was able to identify three different variants of CHRNB4 rs1948

Review of immune responses correlated with COVID-19 outcomes: the fight, debacle and aftermath in the Indonesian context.

In the current pandemic, the highly contagious nature of the severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) leads to an enormous burden for the global health care system and creates challenging socioeconomic problems. Respiratory mucosa, the main entrance of SARS-CoV-2 infection, are equipped with an innate immune defense system as the initial response against infection. Activation of the adaptive immune system facilitates viral clearance as well as providing immunological memory for prevention from subsequent exposure. However, despite repeated efforts at implementing appropriate interventions, severe and fatal cases are continuing to occur and reports of recurrent cases need clarification. Host factors may contribute to the severity of the diseases while viral immune evasion is a common phenomenon leading to severe outcomes and recurrent infection. Discussions of immunological-based tests for screening, herd immunity, along with the possible advantages or potentially futile efforts of development of vaccine and alternative immunotherapy have become a part of daily household conversations. In this review, evidence of innate and adaptive immune responses or lack of them, and immunological problems relevant for SARS-CoV-2 will be summarized. Finally, perspectives for future studies especially in the Indonesian population will be sketched

Epidemiologi, Stadium, dan Derajat Diferensiasi Kanker Kepala dan Leher

Head and neck cancer is one of the deadly types of cancer in Indonesia. The main cause of this cancer is the consumption of alcohol and cigarettes. Head and neck cancer attacks the lips, mouth, palate, pharynx and larynx. Studies about head and neck cancer have been carried out in westerncountries, while in Indonesia is still limited. Earlier studies in western countries expressed that head and neck cancer is more common in men than women. Purpose of research were to determine the epidemiology of head and neck cancer in Indonesia related to patient ratio of men and women andthe correlation of stage and differentiation level of head and neck cancer. Data were obtained from the Rumah Sakit Kanker Dharmais-Pusat Kanker Nasional Jakarta and then analyzed descriptively. The data were analyzed came from 36 patients with head and neck cancer. The results showed, menwith head and neck cancer as much as 52,77% and females 47,22%. There were 16 cases with stage IV cancer, 9 with stage III, 8 with stage II and 2 with stage I. There were 6 cases of stage IV cancer with better differentiation, and there were 2 cases of stage II cancer with a poor differentiation. Head and neck cancer is more common in men than women. There was no correlation between the degree of differentiation-stage head and neck cancer. It was influenced by immunity of each person.Keywords: differentiation level, epidemiology, head and neck cancer, stag

Bibliometric Analysis of Basella ssp. as an Antioxidant

The last ten years have seen the discovery of free radicals and their damaging impacts. Increasing exogenous antioxidant intake could reduce the damage caused by oxidative stress. Several plants have been shown to have antioxidant activity, and one such plant is BasellaI. It is high in phytochemicals which can act as antioxidants, and its consumption may help fight free radicals generated by the body. In particular, this plant is essential for stimulating normal wound healing response. To the best of our knowledge, no bibliometric analysis of published data on Basella as an antioxidant has been done. The goal of this study is to conduct a bibliometric analysis of the research on Basella’s antioxidant properties in the Scopus database using the VOSviewer and RStudio tools. There were 56 articles on Basella as an antioxidant according to the bibliometric analysis. The countries with the highest research output was India (27 documents), and the most productive institution was Chiang Mai University (15 documents). The most productive source was the International Journal of Pharmacy and Pharmaceutical Sciences. P. Giridhar had significant significant impact on papers on Basella as an antioxidant (H-index of 5). The most common keywords were “antioxidant” (859 occurrences with 1,340 total link strength) and “Basella alba” (606 occurrences with 1,048 total link strength). Findings from this data suggest the novelties of Basella as an antioxidant

HUBUNGAN KEBIASAAN MEROKOK DENGAN TITER IgA-EBV DIANTARA ORANG SEHAT

BACKGROUND: Epstein Barr Virus (EBV) infects more than 90% of the world population and is classified as oncogenic virus. EBV is associated with various epithelial and lymphoid malignancies including nasopharyngeal carcinoma (NPC). NPC is a rare malignancy throughout most of the world, however a high incidence rate of NPC is well documented in Southeast Asia and Southern China. NPC was classified by World Health Organization (WHO) into 3 types, where type 3 is the most common and strongly associated with EBV infection. Reactivation of EBV is reflected by IgA-EBV response and can be used as specific biomarker for NPC. High IgA-EBV is detected in NPC patients, but high IgAEBV also detected in haelthy individual and may be a risk factor of NPC. Smoking is known to be associated with high levels of IgA-EBV. OBJECTIVE: To observe the association between smoking and IgA-EBV titres on healthy people . METHOD: A Cross sectional design is used in this study. 279 donors at PMI Kodya Yogyakarta and Sleman was included and given questionnaire about demographic data and smoking habits. Blood samples were collected and tested for IgA-EBV using indirect ELISA method. Association between smoking habits and IgA-EBV titres was analyzed using logistic regression with and without adjustment of age and education. RESULT: In total, 56 subjects (20,07%) have IgA-EBV titres above the cut-off value (0,35). Women was not included in this analysis because of the insufficient number of subjects. Ever smoker is associated with high level of IgA-EBV in men (OR=2,02

Development of Tetra-primer Amplification Refractory Mutation System (ARMS) PCR for Detection of CHRNA3 rs8040868

BACKGROUND: Single nucleotide variations (SNV) have been mapped to be associated with several human conditions and diseases. To validate the association between SNV to certain human traits or diseases, a large number of subjects must be included. Thus, in need of a fast, relatively economic, and reliable genotyping method. This can be achieved through the use of tetra-primer amplification refractory mutation system polymerase chain reaction (Tetra-primer ARMS PCR). This study reports strategy to develop Tetra-primer ARMS PCR-based genotyping of CHRNA3 rs8040868.METHODS: The optimization of Tetra-primer ARMS PCR was done through these steps: identification of gene sequence and position of single mutation; designing outer and inner PCR primers; amplification of target gene fragments through PCR by using outer primer; confirming genotype of the PCR product by using sequencing; determining an optimum ratio of outer and inner primer; and determining optimum annealing temperature and cycles for the PCR program. The PCR products were run in 2% gel agarose electrophoresis and visualized under UV illumination.RESULTS: Outer and inner primer ratio of 1:3 with annealing temperature of 64.4°C and 40x cycles was found to be the most optimum condition. Tetra-primer ARMS PCR was able to confirm the results of the DNA sequence of 2 samples, confirming wild-type variants (TT allele) and the heterozygous mutant (CT allele).CONCLUSION: Tetra-primer ARMS PCR was able to genotype rs8040868 of the CHRNA3 gene.KEYWORDS: tetra-primer ARMS PCR, CHRNA3, rs8040868, genotypin

Bibliometric Analysis of

The last ten years have seen the discovery of free radicals and their damaging impacts. Increasing exogenous antioxidant intake could reduce the damage caused by oxidative stress. Several plants have been shown to have antioxidant activity, and one such plant is BasellaI. It is high in phytochemicals which can act as antioxidants, and its consumption may help fight free radicals generated by the body. In particular, this plant is essential for stimulating normal wound healing response. To the best of our knowledge, no bibliometric analysis of published data on Basella as an antioxidant has been done. The goal of this study is to conduct a bibliometric analysis of the research on Basella’s antioxidant properties in the Scopus database using the VOSviewer and RStudio tools. There were 56 articles on Basella as an antioxidant according to the bibliometric analysis. The countries with the highest research output was India (27 documents), and the most productive institution was Chiang Mai University (15 documents). The most productive source was the International Journal of Pharmacy and Pharmaceutical Sciences. P. Giridhar had significant significant impact on papers on Basella as an antioxidant (H-index of 5). The most common keywords were “antioxidant” (859 occurrences with 1,340 total link strength) and “Basella alba” (606 occurrences with 1,048 total link strength). Findings from this data suggest the novelties of Basella as an antioxidant