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The Relationship between Mouth Squamous Cell Carcinoma (MSCC) with HPV Infection and the Presence of p53 &c-myc Mutation

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Journal of Cancer Therapy 2013年第5期4卷 939-943页

作者： Adi Prayitno Elyana Asnar suhartono taat putra Department of Dental and Mouth Disease Faculty of Medicine University of Sebelas Maret Surakarta Indonesia Department of Pathobiology Faculty of Medicine University of Airlangga Surabaya Indonesia.

Introduction: Now the molecular epidemiology is a new experience. It’s was noted that ninety percent of mouth cancers are squamous cell carcinomas and recorded 0.96% year of all cancers in Indonesia. Human papilloma virus (HPV) was implicated in pathogenesis of cancer. As a remark, that mutations of p53 and c-myc are found 50% in cancer. Objective: Aims of this research were to know the relationship between the mouth squamous cell carcinoma (MSCC) with HPV infection, the presence of p53, and c-myc genes mutation. Methods and Material: Tissue biopsy frozen sections from Benign Mouth Squamous Cell (BMSC) and MSCC patients were collected from Mouth and Dental Department of Muwardi District Hospital in Solo—Indonesia. To amplify L1-HPV gene for fixed the HPV etiology, amplified p53 and c-myc genes continued with SSCP analysis and followed with measurement using densitometer to see mutation existence. The collected data were analyzed with Chi Square Test. Results: None of the sample of patients with BMSC with positive HPV showed p53 gene mutation or c-myc gene. From eleven samples obtained from patients with MSCC who were positive HPV showed 18.2% had mutations in the p53 gene and 9.1% had mutations in c-myc gene. The chisquare test was shown to have significant differences between the MSCC with HPV infection and the presence of p53 and c-myc genes mutation. Conclusion: HPV is a risk ingredient for MSCC.

关键词： HPV p53 and c-myc Mutation MSCC

The Expression of CD8+ and MHC-I in Cervical Cancer with HPV Infection

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Journal of Cancer Therapy 2013年第5期4卷 15-18页

作者： Adi Prayitno Elyana Asnar Okid Parama Astirin suhartono taat putra Mathematical and Science Faculty of Sebelas Maret University Surakarta Indonesia Medical School of Airlangga University Surabaya Indonesia Medical School of Sebelas Maret University Surakarta Indonesia

Introduction: The immunity against cervical cancers is many unknown. Major histocompatability complex class I (MHC-I) with the alpha domain has binding site for Cytotoxic T cells (CD8+) to target cells. The objective of this experiment is to know the expression of CD8+ and MHC-I in cervical cancer with HPV infection. Method: DNA was isolated from seventeen sample cervical cancer tissues frozen section. Diagnose related with HPV was made by PCR. Paraffin block of the tissues was cut in thoroughly cleaned cryotome and place in glass plate that covered with poly-elysine. The immuno-histochemistry is done with monoclonal antibody again MHC-I and CD8+ with TSA-indirect method. The power of expression was counted the number of positive cells to express the MHC-I and CD8+ every 100 cells per one view look. Result: The Result of this experiment shows that expression of MHC-I and CD8+ in cervical cancer with HPV infection is in mild category (30% - 70%). Conclusion: Immunity against uterine cancer played by MHC-I and CD8+ is in milt category. The experiment that related with uterine cancer immunology is suggested.

关键词： CD8+ MHC-I HPV Cervical Cancer

Heat Shock Protein 40 (Hsp40) and Hsp70 Protein Expression in Oral Squamous Cell Carcinoma (OSCC)

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Journal of Cancer Therapy 2013年第3期4卷 734-741页

作者： Adi Prayitno Elyana Asnar Okid Parama Astirin Dinar Rosmala suhartono taat putra Departement of Pathobiology Faculty of Medicine University of Airlangga Surabaya Indonesia Department of Biology Faculty of Science and Mathematica University of Sebelas Maret Surakarta Indonesia Department of Dental and Oral Disease Faculty of Medicine University of Sebelas Maret Surakarta Indonesia Department of Ear-Nose-Throat Faculty of Medicine University of Sebelas Maret Surakarta Indonesia

Introduction: As a chaperone, heat shock protein acts as central integrators of protein homeostasis in cell. The form of these functions is to help setting up a complex protein molecular fold (folded protein) in many important settings, such as growth, differentiation, and the ability to live. It has become clear that the control system plays an important role if the folding process fails or an error occurs, causing folding abnormalities and targeted functionality to accumulate. The accumulation of faulty protein folding would harm cells and can result in death. Apparently, there is a correlation between protein folding error with various diseases, such as diabetes mellitus and cancer. Method: We examined protein levels in all samples using Dotblott with monoclonal antibody anti-Hsp40 and anti-Hsp70. Levels of the protein content was read using a densitometer. Modification of Dot Blot was as follows: treatment was conducted with 3 × SSC, added with 20 mL blocking solution, add with total protein samples of 10 mg/ml on nitrocellulose paper, prehybridized, incubated at 70° for 30 seconds, incubated at 70° for 30 seconds with primary antibody anti-Hsp40 or Hsp70 protein and then added with second antibody HRP anti-Hsp40 or Hsp70 protein, treated with 3 × SSC and visualized with TSA HRP, and then administered with streptavidin, biothynil tyramide, and, finally, added with chromogen (DAB) in a confined space. Result: From the analysis of the data using Manova test with Wilk’s Lambda, there were significant differences in the levels of Hsp40 between Benign Oral Lesion (mean 688.31 area) and OSCC (mean 1354.59 area) patients (p 0.070), there was also a highly significant difference in Hsp70 levels between patients who experienced Benign Oral Lesion (mean 529.82 area) and OSCC (mean 1346.32 area) patients (p 0.006). Conclusion: OSCC patients have increased Hsp70 levels, so it is possible that something is going wrong in protein folding. Errors in protein folding result in a

关键词： Chaperone Hsp40 Hsp70 Misfolding OSCC

Hypoxic Preconditioning Improved Neuroprotective Effect of Bone Marrow-Mesenchymal Stem Cells Transplantation in Acute Glaucoma Models

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Journal of Biomedical Science and Engineering 2016年第4期9卷 245-257页

作者： Titiek Ernawati Gatut Suhendro I Ketut Sudiana suhartono taat putra Harjanto JM Sunarjo Agus Turchan Fedik Abdul Rantam Department of Ophthalmology School of Medicine Widya Mandala University Surabaya Indonesia PHC Hospital Surabaya Indonesia Department of Ophthalmology School of Medicine Airlangga University Surabaya Indonesia Dr. Soetomo General Hospital Surabaya Indonesia Department of Pathology Anatomy School of Medicine Airlangga University Surabaya Indonesia Department of Physiology School of Medicine Airlangga University Surabaya Indonesia Department of Public Health School of Medicine Airlangga University Surabaya Indonesia Department of NeuroSurgery School of Medicine Airlangga University Surabaya Indonesia Laboratory of Stem Cell Institute of Tropical Disease Airlangga University Surabaya Indonesia Regenerative Medicine & Stem Cell Centre Airlangga University Surabaya Indonesia Laboratory of Virology and Immunology Department of Microbiology School of Veterinary Medicine Airlangga University Surabaya Indonesia

This study explored the novel strategy of hypoxic preconditioning of Bone Marrow Mesenchymal Stem Cells (BM-MSCs) before intra vitreal transplantation to improve neuroprotective effects of Retinal Ganglion Cells (RGCs) in Acute Glaucoma Models. The methods of this research were isolated mesenchymal stem cells from the bone marrow of adult wild-type Sprague-Dawley (SD) rats. BM-MSCs were cultured under normoxic or hypoxic (1% oxygen for 24 hours) conditions. Normoxic or hypoxic BM-MSCs were transplanted intravitreally 1 week after ocular hypertension induction by acutely increasing IOP to 100 - 120 mmHg for 60 minutes. Rats were killed 4 weeks after transplanted. Apoptosis was examined by tunnel assay and expression Brn3b (Brn3b = RGCs marker) by immunohistochemical analysis of the retina. Results showed that transplantation of hypoxic preconditioning BM-MSCs in acute glaucoma models resulted in a significant apoptosis decreasing (p < 0.05) and an significant increasing in RGCs (p < 0.05), as well as enhanced mor-phologic and functional benefits of stem cell therapy versus normoxic BM-MSCs transplantation. Conclusions: Hypoxic preconditioning enhances the capacity of BM-MSCs transplantation to improve neuroprotective effects of RGCs in Acute Glaucoma Models.

关键词： Hypoxic Preconditioning Transplantation Bone Marrow-Mesenchymal Stem Cells BM-MSCs Glaucoma Neuroprotective

Lymphocytes CD8+ Expression Mean Increases the Immunity against Cancer

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Journal of Immune Based Therapies, Vaccines and Antimicrobials 2014年第4期3卷 37-41页

作者： Adi Prayitno Anita Sari Laila Wahyuningsih Meutia Srikandi Fitria Okid Parama Astirin Ambar Mudigdo suhartono taat putra Department of Pathobiology Faculty of Medicine University of Sebelas Maret Surakarta Indonesia Department of Biology Faculty of Mathematic and Natural Science Sebelas Maret University Surakarta Indonesia Department of Pathobiology Faculty of Medicine University of Airlangga Surabaya Indonesia

Now molecular epidemiology was meaningful. On the surface of the tumor, cells will express its antigen specific as a tumor cell (example: TSA, tumor specific antigen) and can induce cellular immune response. The result of interactions between group antigens with the immune system cells of the body will cause a rise in the expression of lymphocytes CD8+. The aim of this research is to find out differences in the number of lymphocytes CD8+ expression between benign and malignant tissues. This research is a laboratory experiment with the approach of cross sectional. Samples are taken from benign and malignant tissue biopsy of the breast and cervical uterine that were got from anatomical pathology laboratory, period January to February 2004. A technique using random sampling, is sample acquiring 30 benign cancer and 30 malignant of the breast or cervical uterine. To find out the significance of the difference in the number of lymphocytes CD8+ between benign and malignant of breast or cervical uterine, we used a statistical analysis Anova in SPSS for Windows 15.0 program. In this research the number of lymphocytes average in the benign cancer is 2.9667 cells (breast) and 4.2667 cells (cervical uterine), on the other side malignant tissue of 23.8000 cells (breast) and 25.0333 cells (cervical uterine). From the statistical analyses with Anova the number of lymphocytes CD8+ was very significant differences between benign and malignant of the breast or cervical uterine tissue (p < 0.001). The conclusion of this research is that there is a significant increase of the number of lymphocytes CD8+ expression in cancer tissue.

关键词： Cancer Lymphocyte CD8+ Cellular Immune Response

PKC Is a Target to Modulate the Expression of Receptor Mediated Endocytosis (RME) Mice Macrophages BALB/c for Optimizing the Phagocytosis toward Candida albicans

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Journal of Immune Based Therapies, Vaccines and Antimicrobials 2013年第4期2卷 44-48页

作者： Adi Prayitno Elyana Asnar Okid Parama Astirin Anief Nur Artanti Meutia Srikandi Fitria Eva Agustina Perwitasari suhartono taat putra Departement of Pathobiology Faculty of Medicine University of Airlangga Surabaya Indonesia Department of Biology Faculty of Mathematics and Natural Science University of Sebelas Maret Surakarta Indonesia Department of Dental and Oral Disease Faculty of Medicine University of Sebelas Maret Surakarta Indonesia Department of Pharmacy Faculty of Mathematics and Natural Science University of Sebelas Maret Surakarta Indonesia

Introduction: The existence of receptor-mediated endocytosis (RME) means that selectivity and selectivity occurs in capturing macromolecules. Protein kinase C (PKC) which can be expressed by almost all cells are proteins important in signal transduction groove that plays a role in a number of cell activity, e.g. phagocytosis. Aims: The purpose of this study is to determine the expression of RME after modulating the PKC which is characterized by the number of Candida albicans cells attached to the surface of macrophages. Methods: Peritoneal macrophages cultured BALB/c mice are treated with PMA and/or bisindolylmaleimides of providing levels of 5 ng/ml to 100 ng/ml for 10 minutes. Then immediately insert Candida albicans and observe every 30 minutes for 120 minutes. The research design used the same subject. Data collected in the form of number of Candida albicans cells attached to the surface of macrophages are analyzed with ANOVA statistical test (one way) to show the differences between treatments. Results: The test shows statistically significant difference in the number of Candida albicans cells attached to the surface of macrophages after administration of various levels of PMA (p 0.001). The higher level of PMA is given, the more active the PKC is, the more RME are formed, the more Candida albicans cells attached to the surface of macrophages. Another result shows statistically significant difference in the number of Candida albicans cells attached to the surface of macrophages after administration of various levels of bisindolylmaleimides (p 0.001). The higher level of bisindolylmaleimide is given, the less active PKC is, and the less RME are formed, the less Candida albicans cells attached to the surface of macrophages. Conclusion: Research shows that activator PKC (PMA) can increase the expression of RME on macrophages. Another research shows that inhibitor PKC (bisindolylmaleimides) can decrease the expression of RME on macrophages.

关键词： PMA Bisindolylmaleimides RME PKC Phagocytosis

The effect of curcumin as an antioxidant on cochlea fibroblasts in ototoxic rat models

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Journal of Chinese Pharmaceutical Sciences 2018年第12期27卷 847-854页

作者： Tengku Siti Hajar Haryuna Agustinus Hamonangan Winston Purba Farhat Farhat Soehartono taat putra Department of Otorhinolaryngology Faculty of MedicineUniversitas Sumatera Utara Pathobiology Division Department of Anatomic Pathology Faculty of Medicine Universitas Airlangga

Aminoglycosides(e.g. Gentamicin) are ototoxic drugs and widely prescribed due to their effective antimicrobial actions and affordable prices. This study focused on determining protective effect of curcumin against the damage caused by aminoglycosides. We aimed to demonstrate the potential of curcumin as an antioxidant to increase the expressions of superoxide dismutase(SOD) in fibroblasts of cochlea lateral wall in ototoxic rat models. The experiment was conducted with randomized post test-only control group design by using 32 male Rattusnorvegicus adults which received a combination of gentamicin and curcumin with different durations and doses. Then, the rats underwent terminations and immunohistochemical assay to determine the expression of SOD. The rats receiving gentamicin injection showed significantly decreased expression of SOD(P<0.05), and the administration of curcumin before and after the gentamicin injection showed significantly increased expression of SOD(P<0.05). Collectively, we showed that curcumin was an antioxidant against oxidative stress due to ototoxicity evidenced by the expression of SOD.

关键词： Curcumin Antioxidant Gentamicin Superoxide dismutase Preventive Rat Experimental