Tono Djuwantono
Grup Peneliti Sel Punca, Unit Penelitian Kesehatan, Fakultas Kedokteran Universitas Padjadjaran

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Akt–the Mammalian Target of Rapamycin (mTOR) Pathway Inhibition Increases Cervical Cancer Cell Chemosensitivity to Active Form of Irinotecan (SN-38) Septiani, Leri; Hidayat, Yudi Mulyana; Effendi, Yusuf Sulaeman; Djuwantono, Tono; Luftimas, Dimas Erlangga; Faried, Ahmad
International Journal of Integrated Health Sciences Vol 1, No 1 (2013)
Publisher : International Journal of Integrated Health Sciences

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Objective: To investigate the molecular pathway of the cytotoxic effect of SN-38 in human cervical cancer cell lines.Methods: Two human cervical cancer cell lines were treated with various concentrations of irinotecan for 24–72 hours and the sensitivity was analysed using the MTT assay. Apoptosis was further observed through microscopic examinations. The protein expression was determined using Western blot analysis.  Results: CaSki cells demonstrated the highest sensitivity to SN-38, whereas HeLa cells showed the lowest. In cervical cancer cells, SN-38 induced apoptosis through an intrinsic- and extrinsic-pathways. In addition, we showed that SN-38 downregulated the phosphorylation of Akt-mTOR pathways in CaSki cells, but not in HeLa cells. Interestingly, in HeLa cells, which were more suggestive of a resistant phenotype, pre-treatment with LY294002 and rapamycin inhibited activation of Akt-mTOR signaling and significantly enhanced the sensitivity of HeLa cells to SN-38. Conclusions: Irinotecan exerts its anti-neoplastic effects on cervical cancer cells by inducing apoptosis through caspase-cascade. Inhibition of Akt-mTOR, LY294002 and rapamycin, which is targeted to Akt-mTOR pathways, may sensitize irinotecan-resistant cervical cancer cells.Keywords: Akt-mTOR pathways anti-neoplastic drugs, cervix cancer cells, LY294002, rapamycin DOI: 10.15850/ijihs.v1n1.103
MODIFIKASI METODE ISOLASI SEL ENDOTEL PEMBULUH DARAH OTAK (EPDO) TIKUS: TEKNIK DASAR KULTUR SEL PRIMER DI BIDANG NEUROSAINS Faried, Ahmad; Zafrullah Arifin, Muhammad; Sutiono, Agung Budi; Halim, Danny; Djuwantono, Tono; Achmad, Tri Hanggono
Majalah Kedokteran Bandung Vol 42, No 4
Publisher : Faculty of Medicine, Universitas Padjadjaran

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Metode konvensional isolasi sel endotel pembuluh darah otak (EPDO) masih tergolong sulit, sehingga upaya mendapatkan populasi murni sel ini adalah tantangan. Pada penelitian ini dilakukan isolasi endotel dari tikus Wistar dan mencit C57/Bl6, berdasarkan protokol the care and use of laboratory animals, Universitas Gunma, Jepang. Modifikasi metode isolasi adalah menggunakan gradasi bovine serum albumin (BSA), bukan Dextran-70 yang umumnya dipakai, untuk memisahkan sel EPDO yang bersatu menjadi sel EPDO tunggal. Penelitian ini dilakukan di laboratorium sel kultur, Universitas Padjadjaran bekerjasama dengan Universitas Gunma, Jepang, Januari 2008?Juni 2009. Uji hasil isolasi dan karakteristik sel EPDO dilakukan dengan teknik imunofloresen. Ekspresi tight junction ZO-1, menunjukkan sel EPDO membentuk selapis sel utuh, rapat, tidak bertumpuk dan kompak, sesuai dengan karakteristik dinding EPDO. Fenotip sel EPDO dikonfirmasi dengan acethylated LDL, faktor von Willebrand dan CD31. Penghancuran kapiler dengan collagenase/dispase masih menghasilkan populasi sel yang terkontaminasi perisit. Kontaminasi dimurnikan dengan menggunakan puromycin, tingkat pemurnian sel EPDO mencapai 98,3%. Simpulan, teknik modifikasi berhasil mengisolasi sel EPDO tikus dan mencit, tanpa melakukan intervensi genetik. Puromycin dapat digunakan untuk memurnikan sel EPDO. [MKB. 2010;42(4):161?8].Kata kunci: Metode modifikasi isolasi sel EPDO, pembuluh sawar otak, teknik pemurnian Isolation Modified-Method of Mouse-Brain Microvessel Endothelial Cells: Primary Cell Culture Technique in NeuroscienceIsolation method to obtain pure BMVECs is hard to be done consistently and remains a challenge. In this study, we isolated BMVECs from Wistar rat and C57/Bl6 mouse from Japan SLC. All procedures performed according to guidelines for the care and use of laboratory animals of Gunma University, Japan. The modification of isolation method was using bovine serum albumin (BSA) gradation, not Dextran-70 in which generally used, to separate clusters of BMVECs into single cell. This study was done at Universitas Padjadjaran, in colaboration with Gunma University, Japan, January 2008?June 2009. Further,characteristic and purification results were proven by imunofluorescene staining. The results showed that staining of tight junction, ZO-1, formed a monolayer, tightly packed, non-overlapping and contact-inhibited BMVECs, as expected for a vessel wall endothelial. ECs phenotype confirmed by acethylated LDL, von Willebrand and CD31. The digestion of capillaries generated contaminating pericytes. Contamination was purified using puromycin and the results considered satisfactory (98.3%). In conclusion, our modification procedure allows the isolation of primary rat and mouse BMVECs, which form an endothelial-like monolayer in few days. Puromycin can be used for purification of primary rat and mouse BMVECs. [MKB. 2010;42(4):161?8].Key words: Blood brain barrier, isolation modified-method of mouse-BMVECs, purification methods DOI: http://dx.doi.org/10.15395/mkb.v42n4.30
Cancer Stem Cell: Target Baru Obat Antikanker HALIM, DANNY; DJUWANTONO, TONO; AHMAD, TRI HANGGONO
Indonesian Journal of Cancer Vol 4, No 3 (2010): Jul - Sep 2010
Publisher : "Dharmais" Cancer Center Hospital

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Relaps, resistance and metastasis has become prominent problems that oncologists and cancer patients have to dealt with. Various studies have been done previously concluded that there are a subpopulation of cancer cells, identified as cancer stem cell, most likely to be the cause of relaps, resistance and metastasis of cancer. Cancer stem cell is a subpopulation of cancer cells that possess tumorigenicity, hence it can initiate the growth of tumor. Cancer stem cell has been suspected to be originated from normal stem cells reside in mature tissues, or from progenitor cells that gone through some series of alterations on its characteristics, including mutagenic and non-mutagenic changes. As seen in normal stem cells, cancer stem cell is also oftenly found in its inactive state. Therefore, cancer stem cell is not affected when it treated with many chemotherapeutic agents that are targeting cancer cells that proliferate extensively. Eventually, this event leads to the incidence of cancer relaps on cancer patients who already had series of cancer therapy. Based on this knowledge, it can be concluded that the only absolute way to overcome the incidence of metastasis, resistance and relaps on cancer patients, is to targeting cancer stem cell. Therefore, optimization on protocols of cancer stem cell identification and isolation strived continously. Some molecular markers that are oftenly used as a standard on cancer stem cell isolation are CD34, CD44 and CD133. In line with that, isolation methods that are based on sphere formation and the absorption of coloring dye could also be done to obtain cancer stem cell population. This review article would like to explain the nature of cancer stem cell existence, the pathology underlies its formation, characteristics and identification techniques that are commonly used, and challenges that have to be faced by scientists and physicians in order to optimize the application of cancer stem cell theory for the progress of science and patients’ sake.
Effect of Lycopene on Peritoneal Fluid Malondialdehyde Level in Endometriosis Astarto, Nanang W.; Djuwantono, Tono; Yelliantty, .
Journal of Medicine and Health Vol 1, No 1 (2015)
Publisher : Maranatha Christian University

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Oxidative stress is a factor that contribute in pathogenesis of endometriosis. This condition could also implicated in infertility. Lycopene is a carotenoid that has antioxidant activity. Supplementation of lycopene can reduce oxidative stress in peritoneal fluid of women with endometriosis. This study aims to compare the effect of lycopene on malondialdehyde (MDA) levels in peritoneal fluid of women with endometriosis. It is an experimental study on peritoneal fluid from five women with endometriosis. The result show that lycopene supplementation affecting the MDA levels in peritoneal fluid. MDA levels in peritoneal fluid with lycopene is lower than without lycopene. It conclude that lycopene is proved lowering MDA levels in peritoneal fluid of women with endometriosis. Keywords: endometriosis lycopene, malondialdehyde
Embryo Quality: The Most Critical Factor for Pregnancy Rates after day-2, day-3, and day-5 of Embryo Transfer Djuwantono, Tono
Indonesian Journal of Obstetrics and Gynecology Volume. 34, No. 4, October 2010
Publisher : Indonesian Socety of Obstetrics and Gynecology

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Objective: To determine the most critical factor on day-2, day-3, and day-5 of embryo transfer in correlation with pregnancy rates. Method: This research is a retrospective study in Aster Fertility Clinic, IVF program- Dr. Hasan Sadikin Hospital, Bandung, Indonesia. One hundred ninety five women enrolled in an IVF program conducted from March 2006 through November 2009 at the Aster Fertility Clinic. Effect of embryo-transfer day and any other factors (including quality of embryo, oocyte quantity, difficulty of embryo transfer technique, and blood or mucus contamination on the catheter) on pregnancy rate in IVF. Results: The mean age of the pregnant group was 34.65 (SD = 3.91), and the mean of the infertility period was 7.25 years (SD = 3.54). There were no siginificant differences in pregnancy rates in the day-2, day-3, and day-5 groups. The most critical factor influencing pregnancy was the total score for the quality of embryos [p = 0.001; OR (CI 95%) = 1.94 (0.91 - 4.08)]. Otherwise, the day of embryo transfer, oocyte quantity, and difficulties in embryo transfer did not affect the pregnancy rate (p > 0.05). Conclusion: Our study suggests that the total score for the quality of the embryos was the most critical factor for the success rate of pregnancy rather than the day of embryo transfer, oocyte quantity, difficulty of embryo transfer technique, or contamination of blood and mucus on the catheter. [Indones J Obstet Gynecol 2010; 34-4: 175-9] Keywords: day of embryo transfer, in vitro fertilization, quality of embryo total score, oocytes quantity, catheter contamination
Knowledge and attitude of pregnant woman toward umbilical-cord-blood collection Djuwantono, Tono
Indonesian Journal of Obstetrics and Gynecology Volume. 34. No. 2, April 2010
Publisher : Indonesian Socety of Obstetrics and Gynecology

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Objective: Stem cell technology from umbilical cord blood (UCB) has developed fast as health services. UCB could be used in many kinds of disease. Private UCB bank has been operated in many places all over Indonesia. There’s a need for research to evaluate the knowledge and attitude of pregnant woman towards the act of collecting UCB. Method: This is a cross-sectional study base on question-naire given to 163 pregnant women in three private hospitals in the Bandung City from September - November 2009. The result was analyzed with Rank-spearman correlation and Kruskal-Wallis chisquare. Result: Most respondent were 20 - 29 years old pregnant women (54.6%), with educational level of strata-1 (S-1) (41.1%), with monthly income within 5 - 10 million rupiah (35.6%). This is the second pregnancy or more (42.9%), with gestational age less than 24 weeks (61.3%) and variative obstetric history (59.5%). The average level of knowledge was poor and attitude scale was uncertain (3.4). There is a significant correlation between age (p = 0.008%; CI 95%), level of education (p = 0.0001; CI 95%) and knowledge. There is a significant correlation between gestational age and attitude (p = 0.003; CI 95%). Respondent’s level of knowledge also has a significant correlation with attitude (τs = 0.421; p = 0.0001; CI 95%). Conclusion: This study described the respondent’s poor knowledge will influence their attitudes toward UCB collection, so it is necessary for disseminating more information on UCB as an effort to get better research result on UCB. [Indones J Obstet Gynecol 2010; 34-2: 97-100] Keywords: knowledge level, attitudes, collecting UCB, pregnant woman, the Bandung city
Postlaparoscopic GnRHagonist Therapy does not Improve Spontaneous Conception Rates of Women with Endometriosis Tjandraprawira, Kevin D; Djuwantono, Tono
Indonesian Journal of Obstetrics and Gynecology Volume. 5, No. 2, April 2017
Publisher : Indonesian Socety of Obstetrics and Gynecology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (111.049 KB) | DOI: 10.32771/inajog.v5i2.525

Abstract

Objective: To investigate the reproductive benefits of combining laparoscopic surgery with GnRH-agonist hormonal therapy in treating women with endometriosis. Methods: This is a non-randomized prospective analytic study of patients with endometriosis conducted in a private hospital in Bandung during the period of January 2014 to December 2015, whom were later followed up after 12-24 months for assessment of post-surgical reproductive performance. A total of 121 patients were enrolled, 60 of which received post-surgical GnRH-agonist hormonal therapy. Results: We discovered that 56 women (46.3%) eventually achieved spontaneous conception after surgery but upon further analysis, it was discovered that GnRH-agonist played no significantly meaningful role in improving the spontaneous pregnancy rates of these patients (OR 1.539; 95% CI 0.750-3.159; p-value 0.239). Furthermore, even though there was a wide range as to when they achieved conception, those untreated with hormonal therapy tended to conceive far more quickly than those who were (5.91 ± 6.28; 8.56 ± 4.24; p-value: 0.011). Conclusion: Post-laparoscopic GnRH-agonist administration to women with endometriosis does not significantly improve their chances of spontaneous conception. In fact, such administration seems to delay it. [Indones J Obstet Gynecol 2017; 5-2: 87-93] Keywords: endometriosis, GnRH-agonist, laparoscopy
Postlaparoscopic GnRHagonist Therapy does not Improve Spontaneous Conception Rates of Women with Endometriosis Tjandraprawira, Kevin D; Djuwantono, Tono
Indonesian Journal of Obstetrics and Gynecology Volume. 5, No. 2, April 2017
Publisher : Indonesian Socety of Obstetrics and Gynecology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (111.049 KB) | DOI: 10.32771/inajog.v5i2.525

Abstract

Objective: To investigate the reproductive benefits of combining laparoscopic surgery with GnRH-agonist hormonal therapy in treating women with endometriosis. Methods: This is a non-randomized prospective analytic study of patients with endometriosis conducted in a private hospital in Bandung during the period of January 2014 to December 2015, whom were later followed up after 12-24 months for assessment of post-surgical reproductive performance. A total of 121 patients were enrolled, 60 of which received post-surgical GnRH-agonist hormonal therapy. Results: We discovered that 56 women (46.3%) eventually achieved spontaneous conception after surgery but upon further analysis, it was discovered that GnRH-agonist played no significantly meaningful role in improving the spontaneous pregnancy rates of these patients (OR 1.539; 95% CI 0.750-3.159; p-value 0.239). Furthermore, even though there was a wide range as to when they achieved conception, those untreated with hormonal therapy tended to conceive far more quickly than those who were (5.91 ± 6.28; 8.56 ± 4.24; p-value: 0.011). Conclusion: Post-laparoscopic GnRH-agonist administration to women with endometriosis does not significantly improve their chances of spontaneous conception. In fact, such administration seems to delay it. [Indones J Obstet Gynecol 2017; 5-2: 87-93] Keywords: endometriosis, GnRH-agonist, laparoscopy