Hubungan Ekspresi P-53 Dengan Gambaran Klinikopatologi Pada Penderita Karsinoma Penis Yang Dirawat Di Rumah Sakit Sanglah, Denpasar (2001-2005)

Insiden karsinoma penis di daerah Bali, khususnya yang dirawat di Rumah Sakit Sanglah, cukup tinggi. Sampai saat ini, etiologi dari karsinoma penis multifaktorial, termasuk peran Human Papilloma Virus dalam mekanisme di tingkat selular. Penanganan karsinoma penis saat ini cenderung mengarah ke tissue preserving dan peningkatan kualitas hidup penderita, serta masih terjadi kontroversi. Faktor prediksi terhadap rekurensi, progresivitas, dan prognosis masih didasarkan pada faktor prognostik konvensional berupa gambaran klinikopatologi. Hasil penelitian marker molekuler yang memberikan hasil menjanjikan telah melaporkan mengenai overekspresi p-53 dan antigen skuamosa sel karsinoma untuk memprediksi pembesaran nodul, rekurensi, progresivitas, dan prognosis. Untuk menjawab permasalahan tersebut, penulis meneliti hubungan antara ekspresi p-53 dengan gambaran klinikopatologi pada penderita karsinoma penis yang dirawat di Rumah Sakit Sanglah, Denpasar, periode 2001-2005.Studi ini merupakan penelitian observasional analitik yang dilakukan secara potong lintang (cross sectional) terhadap penderita karsinoma penis yang dirawat di Rumah Sakit Sanglah, Denpasar, periode Januari 2001 sampai Mei 2005. Sampel penelitian diambil dari blok parafin. Kemudian dilakukan pengecatan imunohistokimia dan dinilai ekspresi protein p-53. Hasil penilaian tersebut kemudian dihubungkan dengan stadium, diagnosis histopatologi, grading, dan hasil biopsi kelenjar getah Bening. Kemudian dianalisis dengan uji statistik nonparametrik Mann-Whitney U dan tabel 2 x 2 (crosstabs). Kemaknaan ditentukan berdasarkan nilai p< 0,05. Terdapat hubungan bermakna antara ekspresi p-53 dengan stadium klinis tumor (p = 0,030 ) dan hasil biopsi kelenjar getah Bening inguinal (p = 0,040). Sedangkan hubungan antara ekspresi p-53 dengan diagnosis dan grading histopatologi didapatkan hasil yang tidak bermakna.Kesimpulannya, ekspresi p-53 pada karsinoma penis dapat dipakai sebagai salah satu faktor prognostik dan pertimbangan melakukan biopsi kelenjar getah Bening inguinal, sehingga tindakan biopsi menjadi lebih selektif

α-Mangostin Effectively Inhibits Chikungunya Virus Replication in HepG2 Cells

Chikungunya virus (CHIKV) is an arthropod-transmitted Alphavirus endemic to countries in Africa and Asia, including Indonesia, which causes debilitating arthralgia which can last several years. The rapid spread of CHIKV to new areas makes the discovery of antiviral agents a high priority. α-mangostin is a xanthone from mangosteen (Garcinia mangostana) pericarp and has antiviral activity against Hepatitis C and Dengue viruses. We investigated the antiviral activity of α-mangostin against CHIKV in HepG2 cells in pre-, post- and combination treatments compared to the common antiviral medicine ribavirin, as well their cytotoxicity. Our results show dose-responsive reductions in viral titer in all treatment regimes, with post- and combination treatments being more effective than pre-treatment only (IC50 = 7.79, 5.99 and 6.39 µM, respectively), but with poor specificity (SI = 1.39, 1.81 and 1.70, respectively) compared to ribavirin. Neither compound showed a direct virucidal effect. These results suggest α-mangostin effectively inhibits CHIKV replication in this cell line

THE EXPRESSION AND PURIFICATION OF OCTA-ARGININE APOPTIN AND ITS ABILITY TO KILL CANCER CELLS

Objective: In this research, chicken anemia virus apoptin optimized genetically for expression in Escherichia coli and also modified using (His)6 tag, (Arg)8 tag, and HlyA tag intended for purification needs, penetration enhancement, and secretion from bacterial host to the growth media.Methods: The modified apoptin gene was optimized using an Integrated DNA Technology (IDT). The gene (606 bp) then ordered and synthesized by Eurofins. The apoptin gene was expressed using E. coli BL21 CodonPlus as host, in cultivation temperature of 37 °C, and 25 °C and purified using Ni-NTA agarose beads. The addition of (His) 6 tag enabled the apoptin to be purified in only one step by using nickel column. The expression and purification data analyzed qualitatively as well as quantitatively using SDS-PAGE. MTT assay was used to identify the antitumor effect of octa arginine-apoptin to two kinds of cancer cells, cervix HeLa cancer cell and colon Widr cancer cell. The viability of cell was analyzed when the cell incubated in the variation concentration protein for 72 h.Results: The constructed apoptin gene were expressed in E. coli successfully. The MTT assay indicated that Octaarginin-Apoptin was able to induce apoptosis of HeLa and Widr cells lines in a dose-dependent manner. The recombinant apoptin without tagging with octa-arginine, have no ability to induce apoptosis of HeLa and Widr cells lines. Conclusion: This octa arginine-apoptin may in the future allow the development of a therapeutic protein that is able to kill cancer cells specifically

Correlation between Follicular Fluid’s Androgen Level and Fertilization Rate in Poor Responder Patients Undergone IVF: A Prospective Cohort Done in Yasmin IVF Clinic, Jakarta, Indonesia

Introduction Androstenedione (A4), testosterone (T), and dehidroepiandrosterone (DHEA) are known to be involved in folliculogenesis and follicular maturity. Lower follicular androgen levels in poor responder due to malfunctioned granulosa and theca cells and decreased inhibin B production will decrease FSH and LH. However, androgenic follicular fluid might also induce follicular atresia, decreased oocytes viability, thus affecting fertilization. The aim of current study is to find the correlation between intra-follicular androgen levels and fertilization rate, specifically in poor responder whom contributed in 84% cancelled cycle.Material and Methods This prospective cohort study was done at Yasmin IVF Clinic, Jakarta, Indonesia, in January 2014-March 2015. Infertile women undergone IVF were asked to participate, grouped into poor responder and other, and the androgen levels in the follicular fluid of each consenting patients were measured.Results From total 62 patients, aged 23-47 years old (37.6±5,068), there were 21 poor responders, whereas the other 41 patients with other indication. In poor responder group, levels of androstenedione, testosterone, and DHEA were 50.8 – 272.3 (103.5 ± 59.9), 383.2 – 1747.9 (1114.4 ± 373.2), 11.3 – 454.8 (151.3 ± 96.8), whereas in other group the androgen levels were 44.3 – 326.8 (95.1 ± 61.2), 414.1 – 1463.7 (976.9 ± 258.4), 44.6 – 265.8 (132.7 ± 61.3) with the correlation with fertilization rate 0.609, 0.095, and 0.361 respectively. Conclusion Correlation between follicular androgen levels and fertilization rate found to be not significantly different. The low number of subjects might cause this result, as well as the presence of bias, e.g.male factor and endometriosis might also affect fertilization. A multi-center study with larger sample size added with thorough analysis is needed to reconfirm current data

Effect of Ethylene Glycol on Structural Integrity at Each Stage of Preantral Follicle Development Post Vitrification of Rat Ovary-Histological Analysis

The structure of follicular tissue affects the ability to maintain the structural integrity of follicles against cryoinjury post-vitrification. Histological analysis was conducted on the structural integrity of each stage of preantral follicles post-vitrification using 7.5% and 15.0% doses of ethylene glycol (EG), and ovarian sections with HE staining were observed using an Olympus CX21 microscope connected to Optilab 3.0 lens and Image Raster software. Analysis was conducted on the ovarian cortex in the tracing line area using polygon measure tools to obtain follicle density (follicles/mm2) and follicle index (%) data. The result showed that the EG group 7.5% (KP1) increased follicle density compared to the vitrified group (KKV) in primordial (15.83±1.77) and primary (22.94±8.51) stages. Meanwhile, KP2 (EG 15%) was in primordial (41.92±6.45), primary (11.69±1.95), secondary (33.48±3.63), and tertiary (5.93±0.69) stages. KP1 increased grade 3 follicle index compared to KKV in primary (27.66±2.34), secondary (32.41±6.99), and tertiary (25.00±5.00) stages. Meanwhile, KP2 was in primary (26.87±6.68) and tertiary (25.00±5.00) stages. Both doses of 7.5% and 15.0% EG were able to maintain structural integrity at certain stages of preantral follicles. Secondary and tertiary follicles are the best stages in maintaining grade 3 follicular integrity with the addition of 7.5% EG

Effect of Oral Administration of Dehydroepiandrosterone on PCOS-Like Phenotype of Female C57BL/6 Mice

We aim to evaluate the efficacy and optimal dose of orally administered DHEA in the PCOS mice model by assessing their ovarian morphology and serum FSH, LH, and testosterone levels. Female C57BL/6 mice were divided into a control group (n=5, received daily injections of 0.2 ml sesame oil) and an experimental group, which was further classified into 1) D-45 group (n=5), 2) D-60 (n=5), and 3) D-90 group (n=5) that were treated with 45, 60, and 90 mg/Kg body weight of oral DHEA. After modelling, mice in the control group had a regular estrous cycle, while mice in all experimental groups had a disturbed estrous cycle. Ovarian histology showed several growing follicles and some corpora lutea (CL) in the control, D-60, and D-90 groups. However, some large antral follicles and decreased CL were observed in the D-45 group. Serum FSH was significantly lower in the D-45 group than in the control group (3.73 ± 0.12 vs. 5.28 ± 0.31 mIU/mL, P&lt;0.01), but D-60 and D-90 groups had a similar FSH level to the control group (P&gt;0.05). The serum level of LH and testosterone were significantly elevated in the D-45 group than in the control group (2.52 ± 0.43 vs. 1.30 ± 0.33 mIU/mL, P&lt;0.01 and 1.80 ± 0.32 vs. 1.24 ± 0.23 ng/mL, P&lt;0.01, respectively). Still, the level of LH and testosterone in the D-60 and the D-90 groups was comparable to the control group (P&gt;0.05). Our study suggests that oral administration of DHEA is efficacious in establishing PCOS-like phenotype in mice with the suggested optimal dosage of 45 mg/Kg body weight

The impact of late follicular progesterone level on in vitro fertilization-intracytoplasmic sperm injection outcome: Case-control study

Background: Studies have been conducted to improve the pregnancy rate through the in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) program. In recent years, researchers have been focusing on finding impact of high progesterone level on endometrial receptivity. However, data on whether progesterone level also affects the quality of the embryo is still limited. Objective: The aim is to assess the effect of late follicular progesterone level on the outcome of in vitro fertilization-intracytoplasmic sperm injection (IVF-ICSI). Materials and Methods: This was a case-control of 245 women who underwent in vitro fertilization cycle at Halim Fertility Center, Indonesia. The outcomes assessed were number of oocytes retrieved (OR), maturation rate (MR), fertilization rate (FR), number of good embryos (GE), number of fair embryos (FE), and number of poor embryos (PE). The progesterone (P4) and estradiol (E2) levels were analyzed on the day of human chorionic gonadotropin injection. Serum progesterone level was divided into three groups: 1. low progesterone (≤ 0.50 ng/ml), 2. normal progesterone (0.51-1.50 ng/ml), and 3. high progesterone (&gt; 1.50 ng/ml). All outcomes were compared amongst the groups. Results: Significant differences occurred between progesterone level on the day of human chorionic gonadotropin administration. The number of OR in group 1, 2, and 3 were 8.41 ± 5.88 vs. 12.99 ± 8.51 vs. 17.58 ± 9.52, respectively. Conclusion: Progesterone level on the day of human chorionic gonadotropin injection may have an impact on the outcome of IVF-ICSI. Key words: Intracytoplasmic sperm injection, Embryo, Progesterone, In vitro fertilization