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ILE655VAL Genotyping Study of HER2 - Positive Breast Cancer of Patients from Padang - Indonesia With High Resolution Melting Technique

ILE655VAL Genotyping Study of HER2 - Positive Breast Cancer of Patients from Padang - Indonesia With High Resolution Melting Technique

Dwi Wulandari, Azamris Azamris, Isna Nurhayati, M Ali Warisman, Bugi Ratno Budiarto, Desriani Desriani

Annales Bogorienses Vol. 21, No. 2: 69-75

her2Trastuzumab has proven to be a great improvement in the treatment of HER2+ breast cancer patients, but it is associated with relevant adverse cardiac events and significantly elevated cost of treatment. One of the risk factor for cardiotoxicity due to trastuzumab is the I655V HER2 polymorphism (GTC> ATC mutation) in which the allele mutant (Ile/Val or Val/Val) has a higher risk than the wild type (Ile/Ile). The detection of specific alleles is very important for therapeutic decision-making. In this study, our group has developed a high resolution melting (HRM) with EvaGreen dye method to discriminate specific allele of I655V HER2 (wild type, heterozygote mutant or homozygote mutant) in 66 frozen section samples of HER2+ breast cancer patients. Our study revealed that the wild type is the most prevalent allele (77,27%), whereas heterozygous mutation is significantly present in this research (21.21%) and around 1.52% of samples were detected as minor allele. Only one sample was detected as a minor allele (Val/Val) and may have relatively low abundance in the population. This method has been compared to Sanger sequencing and shows 100% of validity.

Keywords: Breast cancer, HER2, I655V, HRM, allele

Katalog: http://http://jurnal.biotek.lipi.go.id/index.php/annales/article/view/310/pdf

Disusun oleh: Ludya AB/Pustakawan

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Pectinase Production and Clarification Treatments of Apple (Malus Domestica) Juice

Pectinase Production and Clarification Treatments of Apple (Malus Domestica) Juice

Cocok Ana Maryani B, Fahrurrozi, Anja Meryandini

Annales Bogorienses Vol. 21, No. 2: 63-68

Pectinases are a group of enzymes that break down pectin, a polysaccharide that is found in plant cell walls. Today, the application of pectinolytic enzymes plays an important role in food technology for the maceration of fruits and vegetables, including for the extraction and clarification of juice. This research aimed to produce pectinase enzyme for clarifying apple juice. A microbial culture was selected from cocoa bean fermentation samples and identified as Bacillus sp.. Citrus pectin (1%) as the carbon source and peptone (0.1%) as the nitrogen source was found as the best component for pectinase production. The optimum condition of pectinase activity was observed at pH 5 and temperature 40 °C. Enzyme stability studies were performed by incubating crude extract at and the crude enzyme at 40 °C and the percentage activity decrease after one hour storage. Apple juice was treated with the enzyme at different concentrations (0%, 0.5%, 1%, 2%, 4%). Apple juice clarification was evaluated for its percent clarity and viscosity. The result showed that enzyme treatment at 4% in apple juice promoted juice clarification and decreased pH value. In conclusion, the quality of apple juice can be improved by enzymatic treatment using pectinase.

Keywords: Bacillus, clarification, apple juice, pectinase

Katalog: http://http://jurnal.biotek.lipi.go.id/index.php/annales/article/view/311/pdf

Disusun oleh: Ludya AB/Pustakawan

 

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Improvement of HER2I655V TARMS-PCR Performance by DNA Quality Analysis

Improvement of HER2I655V TARMS-PCR Performance by DNA Quality Analysis

Bugi Ratno Budiarto, Azamris, and Desriani

Annales Bogorienses Vol. 21 (2): 52-62.

 tarmReliable TARMS-PCR is a prerequisite in constructing a solid conclusion in genetic diagnostics. The validity of data generated by this molecular technique in most cases is hampered by a false positive result. In attempt to develop a TARMS-PCR for HER2I655V genotyping with no interfering of bias we used DNase I to eliminate DNA contaminant resided in PCR reagent. TARMS-PCR with no enzymatic pretreatment on PCR master mix kit produced a false positive result on HER2I655V TARMS-PCR using as recombinant plasmids system model proven by the presence of multiple PCR products in Non-Template Control (NTC). A dose of 0.1 U of the enzyme could eliminate this DNA contaminant effectively, although this pretreatment altered the specificity of HER2I655V TARMS-PCR genotyping on certain genotype. Combination of touchdown TARMS-PCR with another allelespecific primer recovered specificity of detection on this model system. Interestingly, this optimized HER2I655V TARMS-PCR can only be used for genotyping the clinical samples if only further optimization was done using genomic DNA as template.

Keywords: TARMS-PCR, HER2I655V, DNase I, Polymorphism

Katalog: http://http://jurnal.biotek.lipi.go.id/index.php/annales/article/view/308/pdf

Disusun oleh: Ludya AB/Pustakawan

 

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