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Establishment of human sperm-specific voltage-dependent anion channel 3 recombinant vector for the production of a male contraceptive vaccine Asmarinah, Asmarinah; Bachtiar, Endang W.; Malik, Amarila; Rahayu, Susan
Medical Journal of Indonesia Vol 21, No 2 (2012): May
Publisher : Faculty of Medicine Universitas Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (904.243 KB) | DOI: 10.13181/mji.v21i2.479

Abstract

Background: The aim of this study was to construct a recombinant vector of human sperm specific VDAC3 gene for production of VDAC3 antibody, which is potential as male contraception vaccine.Methods: Target fragment sequence of VDAC3 gene was obtained through amplification of human sperm VDAC3 cDNA with primers covering exon 5 to exon 8. Its PCR product in size of 435 bp was cloned to the pET101/D-TOPO expression vector (5753 bp). E. coli bacteria were transformed with this vector. Cloning of VDAC3 fragment gene to the vector was confirmed by the using of XbaI restriction enzyme and PCR colony method with primers covering exons 5-8 of the human VDAC3 gene.Results: Alignment analysis of amplified fragment covering exon 5 to exon 8 of VDAC3 gene showed 94% homology to human VDAC3 gene from databank. After cloning to the expression vector and transformation to E. coli competent cells, twelve colonies could grow in culture media. Gel electrophoresis of sliced VDAC3 recombinant vector showed a single band in the size of 6181 bp in 8 colonies. After application of PCR colony and amplicon sequencing, the result showed a single band in the size of 435 bp and fragment sequence with 94% identity to human VDAC3 gene.Conclusion: The construction of human sperm specific VDAC3 gene recombinant vector was established in this study. In the future, this recombinant vector will be used to produce VDAC3 antibody for the development of a male contraception vaccine. (Med J Indones. 2012;21:61-5)Keywords: Contraception, recombinant vector, sperm, VDAC3
Characterization of Trypsin-Like Protease of Lactobacillus plantarum FNCC 0270 Trismilah Margono; Wahono Sumaryono; Amarila Malik; Mohamad Sadikin
HAYATI Journal of Biosciences Vol. 21 No. 2 (2014): June 2014
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1468.621 KB) | DOI: 10.4308/hjb.21.2.87

Abstract

Trypsin is an enzyme that has a unique mechanism of cutting peptide bonds specifically at the carboxyl side of lysine or arginine amino acids, with another amino acid. This study aims to analyze a trypsin-like protease (TLP) found in Lactobacillus plantarum FNCC 0270,  by performing partial proteomic tests, i.e. MALDI-TOF/TOF, and standard bioinformatics tools. SDS-PAGE analysis showed 4 protein bands. Two bands of the (P1 and P2) showed molecular weights equivalent to 47.35 and 38.42 kD, each generating 8 and 11 peptide fragments respectively. According to information in www.ncbi.nlm.nih.gov/genbank/structures, the structure  of serine protease HtrA (subs. plantarum L. plantarum ST–III) consists of three domains. Using Clone Manager® software by aligning two sequences we obtained eleven. The Lactobacillus produces of the trypsin-like serine protease has 40-90% similarity. Using the Clustal W2 software we passed the 11 sequences through multiple alignments, and found that the isolate L. plantarum is closely related to L. buchneri, L. brevis, and L. malefermentans on the phylogenetic tree. Alignment analysis results showed that all 8 peptide fragments of band 1 and 11 peptide fragments of band 2,  of the SDS-PAGE, were located in the active domain region of  the fourth trypsin-like serine protease producing Lactobacilli.
Sucrase Activity and Exopolysaccharide Partial Characterization From Three Weissella confusa Strains Amarila Malik; Sheilla Sheilla; Wangi Firdausi; Tri Handayani; Endang Saepudin
HAYATI Journal of Biosciences Vol. 22 No. 3 (2015): July 2015
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (907.029 KB) | DOI: 10.4308/hjb.22.3.130

Abstract

Exopolysaccharides (EPSs) produced by lactic acid bacteria have been well known for their important economic value in food, pharmaceutical and health industries. Large extracellular enzyme sucrases are used by lactic acid bacteria to polymerize EPS, i.e. fructansucrase and glucansucrase. This study aimed to characterize sucrase activity of three Weissella confusa strains MBF8-1, MBF8-2 and MBFCNC-2(1), which were isolated previously from local beverages and their EPS products as well. All strains showed ability to form mucoid and slimy colonies by visual inspection on agar plate using raffinose as substrate suggesting that they possessed fructansucrase activity besides glucansucrase. Obtained EPS products were characterized by HPLC analysis after hydrolysis using 3% TCA at 100C for 1 hour, and by viscosity as well. All strains exhibited similar peak patterns, assuming that all of them possessed fructan EPS product. Supernatant and cell pellet were also analyzed by in situ activity assay performing periodic acid Schiff staining after polyacrylamide gel electrophoresis; only cell pellet showed sucrase activity. Viscosity observation showed that EPS products from all strains were able to increase the viscosity slightly.
Profiling Skin Microbiome in Healthy Young Adult Representing Javanese, Papuans, and Chinese Descent in Indonesia Stella Vania; Amarila Malik
HAYATI Journal of Biosciences Vol. 28 No. 4 (2021): October 2021
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.28.4.249-261

Abstract

Skin serves as the first physical barrier and biological barrier by the colonization of commensal bacteria to prevent pathogen invasion. It was known that the disruption on normal commensal microbiota composition or dysbiosis causes skin diseases, while the skin microbiota diversity itself is influenced by several factors, one of them is ethnicity. This study shows the influence of ethnicity factor in Papuans, Javanese, and Chinese descent young adults living in Jakarta on skin microbiome profiles. The microbiota genomic DNA are extracted from the face skin samples and sequenced with Next Generation Sequencing method to be further analyzed. The result shows that individuals with the same ethnic background share similar skin microbiome characteristics. The greatest skin microbiome alpha diversity is shown by the Papuans and the Chinese descent the smallest. Ethnicity factor that shows statistically significant differences in interindividual dissimilarities are independent of other intriguing factors such as age, geographical location, etc. Therefore the ethnic origin of individuals especially from three ethnics above is a factor to be considered in skin microbiome research and the skin microbiota composition can be used for potential future applications.
Construction of recombinant sox2-encoding plasmids that regulate pluripotency of breast cancer stem cells from indonesian patient Hanifah Rahmi; Tiodinar Theresia; Amarila Malik; Septelia Inawati W
Pharmaciana Vol 9, No 1 (2019): Pharmaciana
Publisher : Universitas Ahmad Dahlan

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (234.1 KB) | DOI: 10.12928/pharmaciana.v9i1.11919

Abstract

A therapy development with recombinant protein is a new and potential innovation to destroy cancer stem cells (CSCs). Various ways of killing CSCs include the provision of polyvalent anti-protein antibodies that code pluripotency. Therefore, it takes a mixture of Oct-4, c-Myc, Sox2, and Klf4 protein antigens that can stimulate the formation of polyvalent antibodies. This study aimed to construct Sox2 recombinant by identifying the target genes by reverse transcriptase PCR and then arranging their designs to be inserted into the cloning vector pET101/D-TOPO®. The target gene was developed by finding the complete sequences of Sox2 nucleotides on the NCBI GenBank. The growth on LB-ampicillin agar plates was amplified by PCR to obtain colonies with pET101/D-TOPO® vectors and inserts of the pluripotent gene of CSCs, then the PCR results were observed through electrophoresis. A total of fifteen colonies have DNA bands with a base pair of about 300 bp in length. The recombinant clones produced Sox2 genes with a base length of 330 bp.
Angka Kejadian dan Faktor yang Memengaruhi Potensi Interaksi Obat dengan Obat pada Pasien Leukemia Akut Anak yang Menjalani Rawat Inap Sri Wulandah Fitriani; Rina Mutiara; Amarila Malik; Murti Andriastuti
Sari Pediatri Vol 18, No 2 (2016)
Publisher : Badan Penerbit Ikatan Dokter Anak Indonesia (BP-IDAI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (98.187 KB) | DOI: 10.14238/sp18.2.2016.129-36

Abstract

Latar belakang. Risiko terjadinya potensi interaksi obat dengan obat (PIOO) pada pasien leukemia akut akibat pemberian terapi multidrug cukup tinggi.Tujuan. Mengetahui angka kejadian dan faktor yang memengaruhi PIOO pada subjek penelitian.Metode. Penelitian dilakukan dengan metode potong lintang. Pengambilan data dilakukan secara retrospektif terhadap pasien leukemia akut pada anak yang menjalani rawat inap di RSCM pada Januari – Desember 2014. Identifikasi PIOO dilakukan menggunakan perangkat lunak Micromedex Drug Reax®.Hasil. Didapat 96 subjek yang memenuhi kriteria penelitian. Subjek berpotensi mengalami PIOO 41,6% dengan tingkat keparahan kontraindikasi (4,70%), berat (60,70%), dan sedang (34,60%). Jenis PIOO paling sering adalah deksametason dan flukonazol yang berpotensi meningkatkan paparan glukokortikoid. Terdapat perbedaan bermakna pada faktor usia (p=0,037), lama hari rawat (p=0,000), dan rerata jumlah obat (p=0,000), sedangkan komorbiditas tidak menunjukkan perbedaan (p=0,082).Kesimpulan. Angka kejadian PIOO pasien leukemia akut pada anak yang menjalani rawat inap di RSCM adalah 41,6%. Faktor usia, lama hari rawat, dan rerata jumlah obat berpengaruh signifikan terhadap kejadian PIOO dengan nilai OR 1,8 terhadap pasien berusia >7 tahun, 6,3 terhadap pasien dengan lama hari rawat >7 hari, dan 5,3 terhadap pasien dengan rerata pemberian >4 obat perhari. Sari 
UGT1A1 gene polymorphisms and jaundice in Indonesian neonates Rinawati Rohsiswatmo; Radhian Amandito; Andiani Wanda Putri; Nilam Sartika; Amarila Malik
Paediatrica Indonesiana Vol 59 No 3 (2019): May 2019
Publisher : Indonesian Pediatric Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (390.063 KB) | DOI: 10.14238/pi59.3.2019.150-6

Abstract

Background Uridine diphospho-glucuronocyltransferase 1A1 (UGT1A1) polymorphisms are a risk factor for unconjugated hyperbilirubinemia in neonates. UGT1A1 polymorphisms decrease bilirubin conjugation, thus causing hyperbilirubinemia. A variety of polymorphisms have been reported, with UGT1A1*60 and UGT1A1*6 especially prominent in the Asian population. Hyperbilirubinemia polymorphism studies are lacking in Indonesian populations. Objective To identify UGT1A1*60 and UGT1A1*6 profiles in Indonesian populations of heterogeneous ethnicity. Methods We enrolled 42 jaundiced neonates who were born from January to April 2017 and treated in the Neonatal Intensive Care Unit of our national referral center, Cipto Mangunkusumo Hospital, Jakarta, Indonesia. Genetic mutations *60 of exon 1 and *6 of the promoter region were analyzed by polymerase chain reaction – restriction fragment length polymorphism methods, with DraI and AvaII as restriction enzymes, respectively. Clinical data including total serum bilirubin and racial information were obtained by medical records and interviews with parents. Results There were no homozygous mutations of UGT1A1*6, but 4.8% of subjects were heterozygous. As for UGT1A1*60, 4.8% were heterozygous and 95.2% were homozygous. Racial variations were not observed for UGT1A1*60, while Betawi descendents were found to have many heteroygous forms of UGT1A1*6. Conclusion Polymorphisms of the UGT1A1 gene were found in Indonesian neonates. Some ethnicities also showed increased tendency towards its incidence, such as the heterozygous form of UGT1A1*6.
Optimasi Perolehan DNA Mikrobioma yang Diekstraksi dari Mekonium dan Feses Neonatus Prematur untuk diaplikasikan pada Next-Gen Sequencing 16S rRNA Larashintya Rulita; Amarila Malik; Radhian Amandito; Rinawati Rohsiswatmo
JURNAL ILMU KEFARMASIAN INDONESIA Vol 19 No 2 (2021): JIFI
Publisher : Fakultas Farmasi Universitas Pancasila

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.35814/jifi.v19i2.1112

Abstract

The composition of the intestinal microbiome of neonates can be identified from meconium and feces by Next-Generation Sequencing (NGS) technology. However, the yield of microbiome DNA of meconium and feces has its own challenges due to the consistency and the high content of PCR inhibitors in these samples. This study aims to optimize the yield of microbiome DNA from meconium sample and feces of pre-term neonates. The DNA yield was obtained by applying certain optimized parameters, i.e., considering the replication and condition of the sample, using a particular kit for DNA extraction, and modifying the DNA elution of the column purification. The genomic DNA obtained was quantified and confirmed using Polymerase Chain Reaction. Results showed that the best DNA yield was achieved by replicating the number of samples twice in the pre-extraction stage, working on fresh meconium and feces samples instead, and suspended the sample in ddH2O prior to extraction process as observed on agarose gel visualization with UV trans-illuminator, as well as in quantitative measurement by a nano spectrophotometer. The best extraction process was using MP Biomedical FastDNA Spin Kit for Soil, in addition to the use of an elution buffer in a smaller volume, resulting in a higher concentration and purity of DNA. In conclusion, we were able to obtain an optimized yet reliable DNA yields, especially from meconium, which fulfilled the quality and quantity requirement for further sequencing process of microbiome.
RNA Therapeutic, Pendekatan Baru Dalam Terapi Gen Malik, Amarila
Majalah Ilmu Kefarmasian Vol. 2, No. 2
Publisher : UI Scholars Hub

Show Abstract | Download Original | Original Source | Check in Google Scholar

Abstract

Some diseases, such as cancer, hereditary and genetic diseases, as well as viral infectious diseases, have been treated unsatisfied by the conventional therapy so far, and even more, by the gene therapy. Together with the pharmaceutical industries, researchers put their best effort to hunt some molecules that can be more favorable for such kind of therapy. After a pivotal study reported in May, 2001, it is certain that Ribonucleic acid (RNA) could effectively silence gene expression in mammalian cell line, so it was then proposed in 2004 the term RNA therapeutics. Antisense RNA therapy which came into the stage earlier seemed to be the one that can answer all the problems in knocking out the unwanted messenger in gene expression. RNA interference (RNAi) concept, which came later in around 2000, began to look like a possible contender. It was reported in some studies that RNAi seems to have some more advantages over both stronger gene-silencing effects and greater ease of use. However, the main obstacle of all kind of gene therapy is, undoubtedly, on the delivery of this molecule to enter the target cell, and mostly, to where it is needed most inside the body. Some studies on genetic material delivery system have been reported, and their progress has been discussed.
Seleksi Galur-Galur Leuconostoc yang Mempunyai Aktivitas Bakteriostatin Terhadap Berbagai Bakteri Indikator Retnaningsih, Agustina; Malik, Amarila; Radji, Maksum
Majalah Ilmu Kefarmasian Vol. 9, No. 2
Publisher : UI Scholars Hub

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Abstract

Lactid Acid Bacteria (LAB) are known to produce bacteriocins which have antimicrobial activity, and possessed to be developed as antibiotic complement. This study aimed to characterize bacteriocins activity from Leuconostoc strains isolated previously from local sources, and to optimize pH and incubation temperature as well. A well diffusion agar assay for zone inhibition method and bacteriocin potency assay performing minimum inhibition concentration (MIC) have been done. Bacterial indicators used in this study are Leu. mesenteroides TISTR 120, and JCM 6124, Staphylococcus aureus FNCC 0047, Listeria monocytogenes FNCC 0156, Escherichia coli FNCC 0183, Pseudomonas aeruginosa FNCC 0063, Salmonella typhi FNCC 0165 and Bacillus subtilis FNCC 0061. Catalase, Trypsin and Protease K were also used for confirmation test. Results revealed that both Leu. mesenteroides MBF2-5 and MBF7-17 possessed bacteriocin activity although against Leu. mesenteroides TISTR 120 and JCM 6124 indicators strains. The optimum pH for bacteriocin potency assay for both Leuconostoc strains MBF2-5 and MBF7-17 was pH 6, whereas the optimum incubation temperature was 32 oc with MIC value of 90% and 80%, respectively.