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All Journal Majalah Ilmiah Biologi BIOSFERA: A Scientific Journal
Hendro Pramono
Fakultas Biologi Unsoed
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Environmental Science

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Karakterisasi Molekuler Ikan Gurami Soang (Osphronemus gouramy Lac.) yang Mati pada Rentang Waktu Berbeda Menggunakan PCR-RFLP Gen Major Histocompatibility Complex Kelas II B Jaka Tri Spetiawan; Agus Nuryanto; Hendro Pramono; Kusbiyanto Kusbiyanto; Petrus H. Tjahja Soedibja
Majalah Ilmiah Biologi BIOSFERA: A Scientific Journal Vol 33, No 2 (2016)
Publisher : Fakultas Biologi | Universitas Jenderal Soedirman

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20884/1.mib.2016.33.2.373

Abstract

Gurami (Osphronemus gouramy Lac.) is a popular fish species among Indonesian people. Several Gurami strains have been cultivated by fish farmer, one of which is Gurami Soang. This strain is belived to have a faster growth rate compared to other strains. However, like other strains, the fingerling of Soang strain have also a low survival and suceptible to disease, especially that caused by Aeromonas hydrophila infection. It has been proved that seeds from a single spawning event show varibale disease resistance. The difference in resistance among individuals is suggested related to the difference in their genetic component. One of the genes responsible for resistance is Major Histocompatibility Complex (MHC) class II B gene. Variability in resistance can be analyzed by using PCR - RFLP technique. PCR-RFLP is a technique that can produce a specific DNA fragments by PCR, followed by cutting the PCR product using restriction enzymes to describe the presence or absence of restriction sites in DNA fragments. This research aims to determine genetic marker to differiantiate between resitant and irresistant individual of Gurami Soang infected by A. hydrophila which die at a different time priod based on PCR-RFLP MHC class IIB gene. The study used survey method with purposive random sampling. The Data of PCR-RFLP band patterns were analyzed descriptively. The result indicated that cutting of the MHC class II B gene using HinfI produce two RFLP bands with 300 bp and 100 bp length in all samples. Meanwhile, the MHC IIB gene was not cuted by PstI, HindIII, BamHI and EcoRI enzymes forall samples. These mean that MHC II gene in all individuals were monomorphic. Therefore,it can be concluded that there is no specific genetic marker to differentiate gurami soang individulas which was dying in different time periods.
Deteksi Resistensi Amoxicillin Helicobacter pylori pada Pasien Dispepsia Indah Sulistiyawati; I Gede Arinton; Hendro Pramono
Majalah Ilmiah Biologi BIOSFERA: A Scientific Journal Vol 33, No 3 (2016)
Publisher : Fakultas Biologi | Universitas Jenderal Soedirman

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20884/1.mib.2016.33.3.308

Abstract

Amoxicillin is one of the antibiotics that commonly used on treatment of H. pylori infection. pbp1A gene mutation in H. pylori is a dominant cause of amoxicillin resistance. This study was aimed to evaluate the presence of H. pylori in patients with dyspepsia by using non-invasive method i.e. H. pylori stool antigen (HPSA) and invasive method i.e. pbp1A gene amplification, and also evaluate the amoxicillin resistance of H. pylori by assessing the pbp1A gene mutations. The samples were  26 faeces and 26 gastric biopsies of patients with dyspepsia from the Internal Disease of Prof. Dr. Margono Soekardjo Hospital in Purwokerto. DNA amplification performed by using polymerase chain reaction (PCR) to detect the presence of amoxicillin resistance encoding gene i.e. penicillin binding protein (pbp1A). Sequencing of the DNA sample was performed at the First Base Malaysian Company, to analyze the existence of a point mutation. DNA sequencing analysis of 12 samples showed the presence of a mutations in pbp1A gene from 2 samples, in the third motive of pbp i.e. amino acid changes, Alanine 599 substituted to Threonin and Threonin 592 to Alanine. Those mutations become a dominant risk factor for resistance of H. pylori, toward the bacterial peptidoglycan synthesis. In this research, it was known that the detection of H. pylori infection by using PCR remains more accurate and specific method. The presence of H. pylori mutant strains in this study may becomes the risk factors of resistance to amoxicillin treatment.     
Karakteristik Morfologi Mycobacterium tuberculosis yang Terpapar Obat Anti TB Isoniazid (INH) secara Morfologi Widodo Widodo; Agus Irianto; Hendro Pramono
Majalah Ilmiah Biologi BIOSFERA: A Scientific Journal Vol 33, No 3 (2016)
Publisher : Fakultas Biologi | Universitas Jenderal Soedirman

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20884/1.mib.2016.33.3.316

Abstract

INH mempunyai peranan dalam menghambat sintesis asam mikolat yang merupakan komponen pembentuk dinding sel M. tuberculosis. Mengetahui pengaruh pemaparan obat INH terhadap karakteristik M. tuberculosis secara morfologi sel. Metode penelitian eksperimen dengan menggunakan Rancangan percobaan acak Lengkap dengan variasi waktu pemaparan 24 jam, 48 jam, 72 jam dan 96 jam pada konsentrasi INH 0,5 μg/ml dengan jumlah ulangan 6. Hasil penelitian menunjukkan ada pengaruh pada masing-masing perlakuan INH 0,5 μg/ml terhadap karakteristik M. tuberculosis, pada pemaparan 24 jam diperoleh persentase pertumbuhan 18,25%,  ukuran rerata sel 1,64 x 0,35 µm, persentase kerusakan sel 28,1%. Pada pemaparan 48 jam diperoleh persentase pertumbuhan sel  7,9%,  ukuran rerata sel 1,64 x 0,36 µm, persentase kerusakan sel 41%, pada pemaparan 72 jam diperoleh persentase pertumbuhan sel 7,0%,  ukuran rerata sel 1,64 x 0,32 µm, persentase kerusakan sel 53,4%, pada pemaparan 96 jam diperoleh persentase pertumbuhan sel 1,6%, ukuran rerata sel 1,61 x 0,29 µm,  persentase kerusakan sel 57%. Perubahan morfologi sel yang terpapar INH 0,5 μg/ml pada permukaan sel terjadi kerutan dan penyusutan panjang sel setelah regenerasi, permukaan sel mulai kasar dan sedikit ada kerusakan, sel mengalami lisis dan sebagian sel berbentuk oval , sel mengalami perubahan bentuk menjadi kokus. karakteristik M. tuberculosis yang terpapar INH 0,5 µg/ml mengalami perubahan morfologi sel, menjadi lebih pendek dan perubahan bentuk dari batang menjadi kokus, namun perubahan morfologi ini tidak terjadi secara permanen pada generasi berikutnya.Kata kunci       : M. tuberculosis, INH, Morfologi, 
Ekspresi mRNA LMP2A Epstein-Barr Virus dari Biopsi Jaringan dalam Blok Parafin Berpotensi sebagai Biomarka dalam Diagnosis Karsinoma Nasofaring Roni Afriansya; Hendro Pramono; Hidayat Sulistyo; Daniel Joko Wahyono
Majalah Ilmiah Biologi BIOSFERA: A Scientific Journal Vol 33, No 3 (2016)
Publisher : Fakultas Biologi | Universitas Jenderal Soedirman

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20884/1.mib.2016.33.3.323

Abstract

Karsinoma nasofaring (KNF) merupakan tumor epitel yang terletak di nasofaring dan merupakan penyakit genetik multifaktor yang endemik. Penyebab utama KNF adalah infeksi oleh Virus Epstein-Barr (EBV). Keberadaan EBV pada penderita KNF dapat diketahui dengan ditemukannya DNA EBV dalam spesimen biopsi jaringan penderita KNF. Genom EBV dan ekspresi sebagian dari produk gen laten virus secara konsisten terdeteksi hampir di setiap sel dalam kanker ini, salah satunya adalah gen Latent Membrant Protein (LMP). Aktivitas mRNA EBV lebih mencerminkan patogenesis KNF yang  sesungguhnya dari pada diagnosis serologi dan pengukuran DNA EBV di sirkulasi. Penelitian dilakukan di Laboratorium PA RSUD Margono Seokarjo, Purwokerto/Lab PA Fakultas Kedokteran dan laboratorium genetika/molekuler Universitas Jenderal Soedirman Purwokerto. Analisis Ekspresi mRNA LMP2A EBV dengan teknik one step RT-PCR dan produk RT-PCR (amplikon cDNA) divisualisasi dengan elektroforesis gel agarosa 1%. Hasil ekspresi mRNA LMP2A EBV adalah 27,3% (6 dari 22 sampel). Kesimpulan, metode one step RT-PCR dapat digunakan untuk menganalisis ekspresi mRNA LMP2A EBV dari sampel biopsi jaringan KNF dalam blok paraffin dan hasil positivitas ekspresi mRNA LMP2A EBV sedang, sehingga berpotensi digunakan sebagai petanda biologi molekul diagnosis KNF.
Co-Authors Agus Irianto Agus Nuryanto Daniel Joko Wahyono Hidayat Sulistyo I Gede Arinton Jaka Tri Spetiawan Kusbiyanto Kusbiyanto Petrus H. Tjahja Soedibja Roni Afriansya Sulistiyawati, Indah Widodo Widodo