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Kloning Gen Melanoma Antigen 1 (Mage-1) dari Jaringan Testis untuk mendapatkan Plasmid Rekombinan Mage-1 Mastutik, Gondo; I’tishom, Reny; Hardjowijoto, Sunaryo; Putra, Suhartono Taat
Majalah Kedokteran Bandung Vol 47, No 4 (2015)
Publisher : Faculty of Medicine, Universitas Padjadjaran

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (952.367 KB)

Abstract

Gen Melanoma antigen-1 (Mage-1) diekspresikan oleh sel spermatogonia jaringan testis normal dan diekspresikan 60−80% oleh liver penderita karsinoma hepatoseluler (KH). Ekspresi Mage-1 merupakan penanda untuk diagnosis KH serta prediktor kanker lambung dan kolorektal. Isolasi messenger ribonucleid acid (mRNA) Mage-1 dari jaringan liver penderita KH sulit dilakukan sehingga dilakukan isolasi mRNA Mage-1 dari jaringan yang mengekspresikan Mage-1, yaitu jaringan testis normal. Penelitian ini merupakan penelitian eksploratif yang dilakukan di Lembaga Penyakit Tropis Universitas Airlangga, Agustus 2006–Agustus 2008. Tujuan untuk mengkloning seluruh area koding gen Mage-1 dari jaringan testis pada vektor dan mendapatkan plamid rekombinan Mage-1. Isolasi seluruh area koding gen Mage-1 dilakukan dengan teknik semi-nested polymerase chain reaction (PCR). Seluruh area koding gen Mage-1 diisolasi, kemudian dikloningkan ke plasmid pET101/D-TOPO dan ditransformasikan ke Escherichia coli (E. coli) Top10 untuk mendapatkan plasmid rekombinan Mage-1. Panjang pET101/D-TOPO adalah 5.753pb dan area koding gen penyandi Mage-1 927 bp sehingga total panjang plasmid rekombinan 6.680 bp (5.753+927). Hasil analisis restriksi dengan EcoRV menunjukkan pita 4.230 dan 2.450 (4.230+2.450 = 6.680). Analisis sekuens gen Mage-1 dari testis mempunyai homologi 100% dengan sekuens M77481 serta NM_004988, dan 99% dengan BC01755. Simpulan, berdasarkan hasil analisis restriksi dan sekuens maka diperoleh plasmid rekombinan pETGM/MAGE1-Testis yang mengandung seluruh area koding gen Mage-1 dan dapat digunakan untuk pengembangan kit diagnostik karsinoma.  [MKB. 2015;47(4):199–206]Kata kunci:  Jaringan testis, karsinoma hepatoseluler, kloning, melanoma antigen-1, pET101/D-TOPOCloning of Melanoma Antigen 1 (Mage-1) Gene from Testicular Tissue to Obtain the Recombinant Plasmid Mage-1Melanoma antigen-1 (Mage-1) is expressed by spermatogonia cells of normal testicular tissue and 60−80% is expressed by the liver of hepatocellular carcinoma (HC) patients. Mage-1 expression is a marker for diagnosing HC and predicting gastric and colorectal cancers. Isolation of messenger ribonucleid acid (mRNA) Mage-1 from the liver tissue of HC patients is difficult; therefore, Mage-1 mRNA isolates can be obtained from tissues that express Mage-1 such as normal testicular tissues . This is an explorative research that was conducted at the Institute of Tropical Diseases of Airlangga University during August 2006–August 2008. The aim was to clone the coding sequence of Mage-1 gene from testicular tissues into a vector and to get recombinant plasmid Mage-1. Isolation of the full-length Mage-1 was performed using semi-nested polymerase chain reaction (PCR) which was then cloned into plasmid pET101/D-TOPO and transformed into Escherichia coli (E. coli) Top10 to get recombinant plasmid Mage-1. The length of pET101/D-TOPO was 5,753 bp and Mage-1 was 927 bp. The length of recombinant plasmid was 6,680 bp (5,753+927). Restriction analysis using EcoRV showed 4,230 and 2,450 bp bands (4,230+2,450=6,680). Sequence analyses showed that Mage-1 was 100% homologous with M77481 and NM_004988, 99% homologous with BC01755. In conclusion, according to the results of the restriction and sequences analysis, the recombinant plasmid pETGM/MAGE1-Testis contains the full length coding region of Mage-1 and is useful for developing the hepatocellular carcinoma diagnostic kits. [MKB. 2015;47(4):199–206] DOI: 10.15395/mkb.v47n4.621
CORRELATION BETWEEN BLOOD SERUM PSA LEVEL AND MMP-2 IN PROSTATE ADENOCARCINOMA Rahaju, Anny Setijo; Meidi, Aniek; Mastutik, Gondo; Mustokoweni, Sjahjenny; Mustika, Arifa
Indonesian Journal of Urology Vol 23 No 2 (2016)
Publisher : Indonesian Urological Association

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.32421/juri.v23i2.220

Abstract

Objective: This study aims to prove the correlation between Prostate Specific Antigen (PSA) blood level and Matrix Metalloproteinase-2 (MMP-2) expression in patients with prostate adenocarcinoma. Material & method: Prostate cancer patients’ data from January 2009 to May 2012 were collected at the Department of Pathology, Soetomo General Hospital Surabaya. Data collected included patient medical documents, PSA blood examination, and histopathological examination. Histopathology slides and paraffin blocks of needle biopsies, Transurethral Resection of Prostate (TURP) and radical prostatectomy of prostate cancer patients werere-read, then the samples that met the inclusion criteria were stained by immunohistochemistry using antibodies MMP-2. Results: Data collection was done to obtain data samples of prostate cancer patients in 2009 to 2012 comprising as many as 22 patients between the ages of 52-91 years. Prostate adenocarcinoma in age of 70-79 was found in 8 patients, with a mean age of 68 years. PSA values obtained from medical documents were between 8.6-594.41 ng/ml. Spearman's test performed in this study showed a positive correlation (one-tailed) (correlation coefficient (r) 0431, p < 0.05) between blood PSA level and MMP-2 expression in patients with prostate adenocarcinoma. Conclusion: Blood PSA level correlates positively with MMP-2 expression in prostate adenocarcinoma.
THE ELEVATION OF OSTEOBLAST ACTIVITY IN RAT BONE MARROW MESENCHYMAL STEM CELLS IN OSTEOGENIC MEDIUM EXPOSED WITH MELATONIN IN PHYSIOLOGICAL DOSES Nurma Yuliyanasari; Gondo Mastutik; Suhartono Taat Putra
Folia Medica Indonesiana Vol. 53 No. 1 (2017): JANUARY - MARCH 2017
Publisher : Faculty of Medicine, Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (629.275 KB) | DOI: 10.20473/fmi.v53i1.5489

Abstract

The objective of this study was to analyze the elevation of osteoblast activity in bone marrow mesenchymal stem cells (BM-MSCs) in osteogenic medium by physiological doses of melatonin administration by measuring alkaline phosphatase (ALP) and osteocalcin level.This studyused BM-MSCs from Rattusnorvegiccus femur bone. Rat BM-MSCs were cultured in a-Mem medium, differentiated in osteogenic medium, and administrated melatonin up to 21 days. This study was divided into 4 groups, K0 (control group), K1 (administrated of 25 nM melatonin), K2 (administrated of 50 nM melatonin), and K3 (administrated of 100 nM melatonin). Rat BM-MSCs were characterized CD 45- and CD 105+ marker using imunocytochemistry analysis and stained with Alizarin red after 15 days treatment. ALP and osteocalcin level were measured using ELISA Kit in days 21st.There weren’t differences ofALP level beetwen groups and there are differences ofosteocalcin level between control groups (K0) withK1, K2, dan K3, and beetwen K1 and K2. The conclusion of this study was that there were an elevation of osteoblast activity in rat BM-MSCs in osteogenic medium by physiological doses of melatonin administration characterized by the elevation of osteocalcin level.
Human pappilomavirus genotype in cervical tissue of patients with Cervical Intraepithelial Neoplasia (CIN) 1, CIN 2, and CIN 3 Gondo Mastutik; Rahmi Alia; Alphania Rahniayu; Anny Setijo Rahaju; Renny I’tishom; Suhartono Taat Putra
Majalah Obstetri dan Ginekologi Vol. 24 No. 3 (2016): September - December
Publisher : Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (181.113 KB) | DOI: 10.20473/mog.V24I32016.74-78

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Objectives: to determine the genotype of HPV in patients with precancerous lesions of cervical tissue.Materials and Methods: An observational study with cross sectional study of patients paraffin block CIN1, CIN2, CIN3 was conducted in Dr Soetomo Hospital. HPV DNA was extracted from paraffin blocks, then performed PCR and genotyping of HPV. The sample consisted of 28 patients with cervical tissue paraffin blocks CIN1, CIN2 and CIN3. Patients aged between 26-74 years (standard deviation 10,12).Results: HPV genotypes that infect patients with CIN1 were HPV16 and 18, CIN2 were HPV16 and 52 and CIN3 were HPV16, 67, and combined infection HPV16/67 and HPV52/67. HPV genotypes in a single infection were 26/28 (HPV16, HPV18, HPV52 and HPV67), and multiple infections were 2/28 (HPV16/67 and HPV52/67).Conclusion: The most dominant HPV genotypes infect patients with precancerous lesions of the cervix were HPV16, HPV67, HPV52, and HPV18.
Hypoglycemic Effects of Rosa damascena Mill. Ethanolic Extract on Blood Glucose Levels and Diameter of Langerhans Pancreatic Islets Bilqis Inayatillah; Tamam Jauhar; Gondo Mastutik; Alphania Rahniayu
Indian Journal of Forensic Medicine & Toxicology Vol. 15 No. 2 (2021): Indian Journal of Forensic Medicine & Toxicology
Publisher : Institute of Medico-legal Publications Pvt Ltd

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.37506/ijfmt.v15i2.14679

Abstract

Background: Diabetes mellitus is a chronic disorder caused by elevated levels of high blood glucose(hyperglycemia). This chronic hyperglycemia causes ROS to accumulate and oxidative stress to increase.Rosa damascena is a plant that contains high levels of antioxidants and polyphenols, but this is not widelyknown to the public. Purpose: The aim of this study is to understand the hypoglycemic effects from Rosadamascena on blood glucose levels and diameter of Langerhans pancreatic islets. Method: Thirty Wistaralbino rats (200-225gr) were divided into 6 groups. Group 1 was a normal control (KN), group 2 washyperglycemic (KD), group 3 was metformin 250 mg/kg BW (KM), group 4 was treatment extract 250 mg/kg BW (P1), group 5 was treatment extract 500 mg/kg BW (P2) group 6 was treatment extract 1000 mg/kg BW (P3). All animals in the group were injected with STZ 50 mg/kg BW, except for group 1. Result:Data were analyzed statistically using SPSS- 22 software with the Kruskall-Wallis test (p<0.05). The resultshowed that ethanolic extract of Rosa damascena has decreased blood glucose levels in days-14 (382±21,97) with 250 mg/kg BW (P1) compare with another treatment group. In histological observed there areno significant different between group (p>0.05). This is showing that ethanolic extract of Rosa damascenahas inability to repaired the diameter of Langerhans pancreatic islets. Conclusion: Ethanolic extract of Rosadamascena decreased blood glucose levels but did not repaired the diameter of Langerhans pancreatic islets.
Analisis Ekspresi BRAF dan TERT pada Kasus Papillary Thyroid Carcinoma Berdasarkan Stratifikasi Risiko ATA Cempaka Harsa Sekarputri,; Etty Hary Kusumastuti,; Gondo Mastutik
Majalah Patologi Indonesia Vol 29 No 3 (2020): MPI
Publisher : Perhimpunan Dokter Spesialis Patologi Indonesia (IAPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (463.269 KB) | DOI: 10.55816/mpi.v29i3.444

Abstract

BackgroundPapillary thyroid carcinoma (PTC) is the most common malignancy in thyroid, accounting for more than 80% of all thyroid cancer.Rrecurrences of the disease reach 30% cases. It has been widely observed that BRAF and TERT is associated with aggressivenessbehaviors in human cancer. Both are involved in the pathogenesis of PTC that may be used as targets for new therapies. AmericanThyroid Association (ATA) risk stratification can define the risk of recurrence in PTC. No marker has been found to determine therisk of PTC recurrence. The purpose of this study is to analyzed BRAF and TERT expression in ATA risk stratification system inPTC classic variant samples.MethodsThe methods of this study is an analytical observational study with cross sectional approach, conducted on 56 samples of PTCclassic variant selected from Department of Anatomical Pathology Dr. Soetomo General Hospital between January 2015-December2017 by determined criteria in ATA risk stratification groups of low risk, intermediate risk and high risk. The expression BRAF andTERT observed using immunohistochemical staining of Santa Cruz monoclonal antibody.ResultsThe result for each group based on ATA risk stratification groups of low risk were 13 samples (23.20%), intermediate risk were 25samples (44.62%) and high risk were 18 samples (32.12%).The correlation was assessed with Spearman’s rho correlation test. There is significant correlation BRAF (p=0.004) with a value ofr=0.374 and TERT(p=0.032) with a value of r=0.287 for ATA risk stratification.ConclusionBRAF and TERT expression showed significant correlation to ATA risk stratification in classic variant PTC.
THE MUTATION STATUS OF KRAS GENE CODON 12 AND 13 IN COLORECTAL ADENOCARCINOMA (Status Mutasi Gen Kras Kodon 12 dan 13 di Adenocarcinoma Kolorektal) Gondo Mastutik; Alphania Rahniayu; Anny Setijo Rahaju; Nila Kurniasari; Reny I’tishom
INDONESIAN JOURNAL OF CLINICAL PATHOLOGY AND MEDICAL LABORATORY Vol 23, No 1 (2016)
Publisher : Indonesian Association of Clinical Pathologist and Medical laboratory

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24293/ijcpml.v23i1.1177

Abstract

Kanker kolorektum merupakan salah satu kanker yang tersering di dunia. Target molekuler untuk pengobatan kanker kolorektumyaitu Epidermal Growth Factor Receptor (EGFR) dengan pemberian antibodi monoklonal anti-EGFR. Pemberian pengobatan ini tidakdapat memberikan efek dampak di pasien dengan status gen KRAS bentuk mutan, sehingga perlu dilakukan pemeriksaan status mutasigen KRAS. Telitian berupa deskriptif dengan pendekatan potong lintang yang bertujuan untuk mendapatkan data status mutasi genKRAS kodon 12 dan 13 di pasien adenocarcinoma colorectal. Deteksi mutasi KRAS dilakukan dengan teknik Polymerase Chain ReactionRestriction Fragment Length Polymorphism (PCR RFLP) yang dikonfirmasi dengan sekuensing. Sampel telitian adalah 30 blok parafinyang diperoleh dari Rumah Sakit Dr.Soetomo Surabaya masa waktu Januari-Desember 2013. Setelah dilakukan ekstraksi DNA terdapat21 sampel yang dapat digunakan untuk pemeriksaan lanjutan. Hasil PCR RFLP menunjukkan terdapat 7/21 mutasi pada kodon12 dan tidak terdapat mutasi gen KRAS pada kodon 13. Mutasi pada kodon 12 yaitu GGT>GCT, GGT>GGA dan GGT>GAT yangmenyebabkan perubahan asam amino Gly12Ala, Gly12Gly dan Gly12Asp. Simpulan telitian ini adalah mutasi gen KRAS kodon 12 padaadenocarcinoma colorectal di Rumah Sakit Dr. Soetomo Surabaya sebanyak 33%.
Distribution of Human Papilloma Virus (HPV) in Cervical Adenocarcinoma and Adenosquamous Carcinoma Gondo Mastutik; Alphania Rahniayu; Nila Kurniasari; Anny Setijo Rahaju; Budi Harjanto
Folia Medica Indonesiana Vol. 57 No. 2 (2021): June
Publisher : Faculty of Medicine, Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20473/fmi.v57i2.26473

Abstract

Approximately 20-30% of all cervical cancer cases are adenocarcinoma and adenosquamous carcinoma. Around 70% of all of these types of cancer are related to infection of Human Papillomavirus (HPV). This study evaluated the distribution of HPV genotype in cervical adenocarcinoma and adenosquamous carcinoma. A cross-sectional study was conducted at the Department of Anatomic Pathology, Dr. Soetomo General Academic Hospital, Surabaya, Indonesia, from January to December 2015. The sample were 22 formalin-fixed paraffin-embedded (FFPE) of cervical adenocarcinoma tissues and adenosquamous carcinoma tissues. FFPE was used for DNA extraction and followed with HPV genotyping to detect 40 genotypes of HPV, including low risk (LR) and high risk (HR) HPV. The histopathological types of adenocarcinomas were adenocarcinoma NOS and mucinous adenocarcinoma, while the adenosquamous carcinoma types were adenosquamous carcinoma and adenosquamous carcinoma glassy. All of the specimens were infected by HPV. In cervical adenocarcinoma, the infection was by HPV 6, 11, 16, 18, 31, 45, 68B, and 72, and in adenosquamous carcinoma by HPV 6, 16, 18, 45, and 59. HPV 18 was predominant, which was found in 13/22 (59.1%) in adenocarcinoma and 19/22 (86.4%) in adenosquamous carcinoma. Single infection and multiple infections in adenocarcinoma were 13/22 (59.1%) and 9/22 (40.9%), while in adenosquamous carcinoma were 21/22 (95.5%) and 1/22 (4.5%) respectively. The most common HR HPVs found in this study were HPV 18, HPV 45, HPV 16 and LR HPV are HPV 11, HPV 6.
The Genotype of Human Papilloma Virus of Male Patient with Anogenital Warts Dwi Murtiastutik; Gondo Mastutik; Alphania Rahniayu; Afria Arista; Trisniartami Setyaningrum
Folia Medica Indonesiana Vol. 55 No. 2 (2019): June
Publisher : Faculty of Medicine, Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20473/fmi.v55i2.24581

Abstract

Human papillomavirus (HPV) infection is one of the most common sexually transmitted infections. HPV high risk (HPVHR) were HPV16,18 related with invasive penile carcinomas, and HPV low risk (HPVLR) were HPV6,11 related to anogenital warts. Male infection is usually asymptomatic that it would be explain increasing the incidence of HPV associated cancers. Identification HPV genotype is very important for predicting the development of the diseases, to be benign or malignant cancer. The objective of this study was to identify the genotype of HPV that infect men with anogential warts. This research used 12 biopsy specimens from men patient with anogenital warts at Outpatient clinic of Department Dermatology and Venereology, Dr. Soetomo General Hospital period 2016-2017. The specimens were diagnozed by pathologist and HPV gentoyping was done to detect 40 HPV genotype including HPVHR and HPVLR. The result showed that 58% (7/12) were positive for HPVLR and 42% (5/12) were positive for HPV LR/HR. The genotype HPV that infected men patient with anogenital warts is HPVLR (HPV6,11) and HPVHR (HPV18,51,52,82) with single infection of HPVLR or mutiple infection HPVLR/LR or HPVLR/HR. The infection of HPVHR would be develops to be malignant transformation. It suggested that HPV genotype needs to be checked the for the anogenital warts cases for predicting the development of the diseases.
The Expression of E6 HPV, P53 and P16ink4a at Well, Moderately, and Poorly Differentiated Cervical Adenocarcinoma Gondo Mastutik; Alphania Rahniayu; Nila Kurniasari; Anny Setijo Rahaju; Rahmi Alia; Sjahjenny Mustokoweni
Folia Medica Indonesiana Vol. 55 No. 4 (2019): December
Publisher : Faculty of Medicine, Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20473/fmi.v55i4.24468

Abstract

The objective of this study is to analyze the expression of E6 Humanpapilloma virus (HPV), p53, and p16INK4A in cervical adenocarcinoma grade well differentiated (WD), moderately differentiated (MD), and poorly differentiated (PD). A cross sectional study conducted at Department of Anatomic Pathology, Dr. Soetomo General Academic Hospital Surabaya Indonesia using formalin fix paraffin embedded (FFPE) from cervical normal and cervical adenocarcioma grade WD, MD, and PD. The expression of E6 HPV, p53, and p16INK4A was performed by immunohistochemistry (IHC) staining. Data were analyzed with Kruskal-Wallis and continued with Mann-Withney test. The expression of E6 HPV in the cervical adenocarcinoma showed 35.9% specimens represented negative and 64.1% specimens represented positive. There was no significant difference in the expression of E6 HPV and p53 in cervical adenocarcinoma between grade WD, MD, and PD. The p16INK4A was overexpressed, shown as diffuse appearance in 89.7% of the specimens. There was a significant difference in the expression of p16INK4A between grade WD and MD with PD. In conclusion, some of cervical adecarcinoma were not caused by infection of HPV type 16 or 18 and the expression of p16INK4A might take a role in the developing of malignancy that caused by infection of HPV.