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All Journal Majalah Kedokteran Bandung Mutiara Medika: Jurnal Kedokteran dan Kesehatan
- Mashuri, -
Bagian Radiologi Fakultas Kedokteran Universitas Lambung Mangkurat-RSUD Ulin Banjarmasin,
Health Professions Medicine & Pharmacology

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Modifikasi Protein Akibat Pembebanan Glukosa dengan Model Reaksi Glikosilasi Nonenzimatik in vitro Suhartono, Eko; Setiawan, Bambang; Mashuri, -; Juniarti, Maya; Kamilah, Insanul; Haudhiya, -
Mutiara Medika: Jurnal Kedokteran dan Kesehatan Vol 8, No 1 (2008)
Publisher : Universitas Muhammadiyah Yogyakarta

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Abstract

Glycocylation reaction causes protein modification. Glycocylation is a reaction between aldehyde group from reducing sugar with amine group of protein. The aim of this study was to measure Advanced Glycation End Products (AGEs) formation, dicarbonyl compound and tyrosine degradation in glycocylation reaction in vitro. A quasi experimental study was done to four treated groups, i.e. P1= 5 ml Bovine Serum Albumin (BSA), 10 ml phosphate buffer dan 10 ml aquadest; P2= 5 ml BSA, 10 ml phosphate buffer and 10 ml glucose 125 mM; P3= 5 ml BSA, 10 ml phosphate buffer and 10 ml glucose 250 mM; P4= 5 ml BSA, 10 ml phosphate buffer and 10 ml glucose 500 mM. AGEs compound was measured for 21 days using spectrophotometer at X = 390 nm. Dicarbonyl compound was measured by DNPH odification methods at X = 470 nm. Tyrosine degradation was measured usingMillon-Nasse reaction. Anova and Tuckey HSD test concluded there are significant difference between each groups (P<0,05). Based on correlation regresion test conclude that the increase of dicarbonyl compounds, AGEs and tyrosine degradation had positive correlation with increase of glucose concentration. Glucose overloading could induce protein modification in vitro.Salah satu penyebab modifikasi protein adalah reaksi glikosilasi. Reaksi glikosilasi adalah reaksi antara gugus aldehid gula pereduksi dengan gugus amina protein. Penelitian ini bertujuan mengukur Advanced Glycation End Products (AGEs), senyawa dikarbonil maupun degradasi tirosin pada reaksi glikosilasi in vitro. Penelitian ini merupakan penelitian eksperimental semu denganpre andpost control group design terhadap empat kelompok perlakuan, yaitu P1= 5 ml Bovine Serum Albumin (BSA), 10 ml buffer fosfat dan 10 ml aquadest; P2= 5 ml BSA, 10 ml buffer fosfat dan 10 ml glukosa 125 mM; P3= 5 ml BSA, 10 ml buffer fosfat dan 10 ml glukosa 250 mM; P4= 5 ml BSA, 10 ml buffer fosfat dan 10 ml glukosa 500 mM. Absorbansi senyawa AGEs diukur selama 21 hari pada X = 340 nm sedangkan absorbansi senyawa dikarbonil diukur dengan X = 390 nm dan absorbansi degradasi tirosin dengan k=470 nm. Pengukuran absorbansi senyawa dikarbonil menggunakan metoda DNPH yang dimodifikasi, sedangkan pengukuran degradasi tirosin menggunakan reaksi Millon-Nasse. Berdasarkan hasil uji Anova dan Beda Nyata Jujur, disimpulkan bahwa terdapat perbedaan yang signifikan (P<0,05) tiap kelompok perlakuan. Berdasarkan uji korelasi regresi dapat disimpulkan bahwa pembentukan senyawa dikarbonil, AGEs dan degradasi tirosin berkorelasi positif dengan peningkatan konsentrasi glukosa. Pembebanan glukosa yang berlebih dapat memicu modifikasi protein in vitro.
INTERAKSI ANTIBODI MONOKLONAL NIMOTUZUMAB DENGAN RESEPTOR HER-1 YANG DIEKSPRESIKAN GLIOMA SEREBRI Mashuri, -; Soetikno, Rista D.; Mutalib, A.
Majalah Kedokteran Bandung Vol 45, No 2 (2013)
Publisher : Faculty of Medicine, Universitas Padjadjaran

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Abstract

Human epidermal receptor (HER-1) merupakan anggota famili epidermal growth factor receptor (EGFR) yang banyak diekspresikan glioma. Interaksi HER-1 dengan antibodi monoklonal (Mab) merupakan salah satu pendekatan untuk diagnosis dikaitkan dengan kespesifikan interaksinya yang lebih terarah dalam mencapai target molekul. Penelitian ini bertujuan menganalisis interaksi antara antibodi monoklonal nimotuzumab dan reseptor HER-1 yang diekspresikan glioma serebri. Penelitian menggunakan metode studi analitik korelasional dengan rancangan eksperimental untuk menilai interaksi antibodi monoklonal nimotuzumab (Mab) dengan reseptor HER-1 yang diekspresikan cell-line glioma. Penelitian dilakukan di Laboratorium Pusat Radioisotop dan Radiofarmaka (PRR) Badan Tenaga Atom Nasional, Serpong Tangerang periode Januari?Juli 2012. Subjek penelitian cell-line glioma yang didapat dari American Type Culture Collection (ATCC), diinkubasi dengan sejumlah nimotuzumab dengan konsentrasi yang berbeda-beda secara berturut-turut 0,22; 0,11; 0,055; 0,0275; 0,01375 pM, selanjutnya dilakukan penentuan interaksi nimotuzumab dengan HER-1 secara in vitro menggunakan Formula Scatchard. Nilai interaksi ditunjukkan dengan nilai tetapan disosiasi dan kerapatan reseptor dengan nilai Bmax. Hasil penelitian menunjukkan korelasi kuat antara konsentrasi nimotuzumab dan HER-1 yang diekspresikan glioma serebri (r=0,922;p<0,001). Nilai Kd nimotuzumab didapatkan 6x10-7 M dan nilai Bmax sebesar 1,64x10-5 mol/mg protein. Simpulan, terdapat interaksi antara antibodi monoklonal nimotuzumab dan reseptor HER-1 yang diekspresikan cell-line glioma serebri. [MKB. 2013;45(2):86?90]Kata kunci: Afinitas pengikatan, antibodi monoklonal, glioma, HER-1, nimotuzumabInteraction of Nimotuzumab Monoclonal Antibody with Human Epidermal Receptor-1 Expressed by Cerebral GliomaHuman epidermal receptor (HER-1) is a family member of the epidermal growth factor receptor (EGFR) which is widely expressed in glioma. Interaction of the monoclonal antibody with the HER-1 is a diagnosis approach which is associated with the specificities of a more targeted interactions in reaching the target molecule. This study aims to analyze the interaction between nimotuzumab monoclonal antibody and HER-1 receptor expressed by cerebral gliomas. This study is using correlational analytic studies with an experimental design to assess the interaction of nimotuzumab monoclonal antibody to HER-1 expressed by glioma cell line. This research was conducted in Center for Radioisotopes and Radiopharmaceuticals Laboratory, National Nuclear Energy Agency, Serpong, Tangerang in January?July 2012. Subjects were glioma cell lines obtained from the American Type Culture Collection (ATCC)which were incubated with nimotuzumab in different concentrations, i.e. 0.22, 0.11, 0.055, 0.0275, 0.01375 pM, respectively. Furthermore, the determination of the in vitro interaction of nimotuzumab with HER-1 was conducted using Scatchard formula. The value of the interaction is shown by the value of the dissociation constant and receptor density indicated by the Bmax value. The results showed a strong correlation between the concentration of nimotuzumab with HER-1 expressed by cerebral gliomas (r=0.922, p<0.001). Thenimotuzumab Kd value obtained was 6x10-7 M while the Bmax value was 1.64 x10-5 mol/mg proteins. In conclusion, there is an interaction between monoclonal antibody nimotuzumab with HER-1 expressed by the cerebral glioma cell line. [MKB. 2013;45(2):86?90]Key words: Binding affinity, glioma, HER-1, monoclonal antibodies, nimotuzumab DOI: http://dx.doi.org/10.15395/mkb.v45n2.88
Co-Authors A. Mutalib Bambang Setiawan Eko Suhartono Haudhiya, - Juniarti, Maya Kamilah, Insanul Rista D. Soetikno, Rista D.