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Abstrak :
ABSTRAK
Manifestasi klinis yang menonjol pada demam berdarah dengue adalah kebocoran plasma karena gangguan pada sel endotel vaskular. Anti NS-1 dapat bereaksi silang dengan Protein Disulfide Isomerase (PDI) pada sel endotel. Jambu biji biasa digunakan untuk mengatasi gejala dengue namun belum pernah ada penelitian secara in vitro untuk mengetahui mekanisme senyawa aktif yaitu lycopene yang terdapat di dalamnya. Tujuan penelitian ini adalah untuk mengetahui apakah lycopene dapat menurunkan apoptosis yang ditandai dengan Annexin V dan Heme oxygenase -1 (HO-1). Penelitian ini menggunakan kultur Human Umbillical Vein Endothelial Cells (HUVEC) yang diberi stimulasi anti NS-1 dan terdiri atas 5 perlakuan yaitu kontrol positif, kontrol negatif, dan lycopene dosis 0.5, 1 dan 2 μM. Kontrol positif adalah HUVEC yang diberi anti NS-1 dan basitrasin karena basitrasin sudah diketahui bekerja sebagai anti PDI dan dapat menghambat apoptosis. Kontrol negatif adalah kultur HUVEC yang diberi anti NS-1 tanpa perlakuan lycopene. Hasil penelitian menunjukkan bahwa hanya Annexin V pada kontrol positif menunjukkan hasil yang rendah secara bermakna dibandingkan perlakuan lainnya. Hal ini menunjukkan bahwa lycopene dengan dosis 0.5, 1 dan 2 μM tidak dapat menghambat apoptosis. Kadar HO-1 pada semua perlakuan tidak menunjukkan perbedaan bermakna. Hal ini menunjukkan bahwa baik basitrasin maupun lycopene tidak mempengaruhi metabolisme HO-1. Dapat disimpulkan bahwa senyawa lycopene dengan dosis dosis 0.5, 1 dan 2 μM tidak menunjukkan pengaruh dalam apoptosis sel endotel setelah terjadi infeksi dengue.

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ABSTRACT
Prominent clinical manifestation of dengue hemorrhagic fever is plasma leakage due to malfunction of endothelial cells. There is cross reaction between anti NS-1 and Protein Disulfide Isomerase (PDI) on endothelial cells. Psidium guajava is commonly used to improve condition in dengue symptoms but there is no research yet that study the in vitro mechanism how lycopene, a compound in Psidium guajava, works in this case. So this study aimed to know whether lycopene will decrease apoptosis of endothelial cells marked by Annexin V and Heme oxygenase-1 (HO-1) This study used Human umbillical vein endothelial cells (HUVEC) that given anti NS-1 stimulation and consisted of positive control, negative control and lycopene treatment with 0.5, 1 and 2 μM dose. Positive control is HUVEC with anti NS-1 and bacitracin that known act as anti PDI and inhibit apoptosis. Negative control is HUVEC with anti NS-1. Results showed that Annexin V only in positive control had lower Annexin V significantly compared to other treatments. This showed that lycopene 0.5,1 and 2 μM was not able to inhibit apoptosis. HO-1 in all treatments did not show significant difference. This showed that either bacitracin nor lycopene did not give effect to HO-1 metabolism. It was concluded that lycopene with 0.5, 1 dan 2 μM dose has no effect in endothelial cell apoptosis after dengue infection.

Abstrak :
Tujuan Studi ini adalah untuk mengetahui pengaruh hipoksia terhadap pola ekspresi gen HIF-1α pada jantung tikus serta mengamati timbulnya apoptosis pada kardiomiosit akibat hipoksia sistemik. Metode Hewan coba (tikus Sprague-Dawley) dibagi secara acak menjadi 7 kelompok (n= 4 per kelompok): kelompok kontrol normoksia (oksigen atmosfir), dan beberapa kelompok hipoksia yang ditempatkan dalam sungkup-hipoksik (kadar O2 8%) selama 1, 3, 7, 14, 21, dan 28 hari. Pemeriksaan ekspresi gen HIF-1α dilakukan dengan real-time PCR dan apoptosis dengan metode TUNEL. Hasil Dibandingkan dengan kelompok normoksia, ekspresi gen HIF-1α meningkat secara bertahap sejalan dengan lamanya hipoksia dan mencapai puncak pada hari ke-21. Tidak ada sel yang terlabel dengan cara TUNEL pada kelompok kontrol. Dibandingkan dengan kontrol, indeks apoptotik meningkat sejalan dengan lamanya hipoksia. Tidak ada hubungan bermakna antara peningkatan ekspresi HIF-1α dengan peningkatan indeks apoptotik. Kesimpulan Hipoksia sistemik kronik mengakibatkan peningkatan ekspresi mRNA HIF-1α dan apoptosis pada kardiomiosit.

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Abstract
Aim This study explored the expression of HIF-1α in hypoxic cardiac muscle in mice, and observed the evidence of apoptosis in hypoxia induced cardiomyocyte. Methods Male Sprague-Dawley rats, were randomized into 7 groups (n= 4 per group): control normoxia group that was exposed to atmospheric oxygen and hypoxia groups that were housed in hypoxic chambers (O2 level 8%) for 1, 3, 7, 14, 21, and 28 days respectively. Animals were sacrificed, hearts were rapidly excised, total RNA was extracted with an mRNA isolation kit and the expression of HIF-1α mRNA was then detected by real-time RT-PCR. Apoptosis was assessed by TUNEL method. Results For rat in hypoxia group, the expression of HIF-1α mRNA in cardiac myocytes was clearly up-regulated compared to the control normoxia group. Further, HIF-1α expression level elevated gradually and reached a peak at 21 days of hypoxia. No cell labeled by the TUNEL method was detected in the control group. Compared with the control group, the apoptotic index was significantly increased in the hypoxia group (P < 0.05). There was no significant correlation between the elevation of HIF-1α mRNA and the elevation of apoptotic index. Conclusion Systemic chronic hypoxia caused the elevation of HIF-1α mRNA and apoptosis in cardiac myocytes.

Abstrak :
Mahkota Dewa as a traditional plant has been commonly used as traditional cancer medication. However, the mechanism of usage is not yet clear. The objective of this study was to know the mechanism of the protection effect of Mahkota Dewa on Benzo(a)pyrene (BaP) induced cytotoxicity in CCRF-CEM cell line. The result showed BaP induced cell death with in CCRF-CEM cell line was dose-dependent but not based on time-course. Exposure of this cell for 24 h with variation of dose between 5-20 μM increased the percentage of apoptosis to about 15%. On the other hand, Mahkota Dewa itself has dose-dependently induced cytotoxicity and has no effect in the inhibition of BaP exposure. Phosphorylation of p38 MAPK in both BaP and Mahkota Dewa induced cytotoxicity has been seen but the involvement of oxidative stress is unclear. However, in other cancer cell line SH-SY5Y human neuroblastoma cells, the inhibition efffect of Mahkota Dewa in BaP exposure has been seen and no cytotoxicity effect appeared in this cell line. In conclusion, Mahkota Dewa has induced apoptosis in CCRF-CEM cancer cell line but not in SH-SY5Y cell line, so it has a potential anticancer effect; Mahkota Dewa, however, requires more researches on DNA level using other type of cancer to observe the mechanism.

Efek Penghambatan Mahkota Dewa (Phaleria macrocarpa) pada Sitotoksisitas CCRF-CEM Cell Lines yang Terpajan oleh Benzo(a)pyrene. Mahkota Dewa adalah tumbuhan tradisional yang umumnya digunakan sebagai obat kanker tradisional. Namun belum terdapat kejelasan mengenai mekanisme penggunaannya. Tujuan penelitian ini adalah untuk mengetahui mekanisme efek proteksi Mahkota Dewa pada sitotoksisitas CCRF-CEM cell line yang terpajan oleh Benzo(a)pyrene. Hasil penelitian menunjukkan kematian sel dalam CCRF-CEM cell line yang diinduksi oleh BaP terjadi secara dependen terhadap dosis, tetapi bukan didasari oleh jangka waktunya. Paparan sel ini dibiarkan selama 24 jam dengan dosis bervariasi antara 5-20 μM dan mengakibatkan peningkatan persentase apoptosis sampai sekitar 15%. Di lain pihak, Mahkota Dewa itu sendiri telah menginduksi sitotoksisitas secara dependen terhadap dosis, dan tidak ditemukan efek terhadap penghambatan paparan BaP. Fosforilasi p38 MAPK baik dalam BaP dan sitotoksisitas yang terpajan oleh Mahkota Dewa telah terlihat. Akan tetapi keterlibatan stress oksidatif tidak jelas terlihat. Meskipun demikian, dalam cell line kanker lainnya seperti SH-SY5Y sel neuroblastoma manusia, efek penghambatan Mahkota Dewa dalam paparan BaP telah terlihat dan tidak terdapat adanya efek sitotoksisitas yang muncul di cell line ini. Sebagai kesimpulan, Mahkota Dewa telah menginduksi apoptosis pada cell line kanker CCRF-CEM. Namun apoptosis tidak diinduksi pada SH-SY5Y cell line sehingga tumbuhan ini berpotensi memiliki efek antikanker. Meskipun demikian, perlu lebih banyak penelitian mengenai Mahkota Dewa pada level DNA dengan menggunakan jenis kanker lainnya agar mekanismenya dapat diobservasi.

Abstrak :
Kanker serviks termasuk salah satu jenis kanker dengan tingkat prevalensi kasus yang tinggi di Indonesia. Berdasarkan GLOBOCAN 2020, ditemukan 36.633 (36% dari total kasus kanker pada wanita) kasus kanker serviks baru dan 21.003 kematian. Pengobatan terhadap kanker serviks saat ini masih menimbulkan efek samping seperti resistensi obat pada sejumlah kasus sehingga diperlukan alternatif pengobatan. Salah satu senyawa yang memiliki potensi antikanker, yaitu Eucalyptol. Eucalyptol adalah senyawa utama dalam tumbuhan eukaliptus seperti Eucalyptus globulus. Studi pengaruh konsentrasi eucalyptol terhadap viabilitas dan deteksi apoptosis sel HeLa dilakukan dengan konsentrasi 25, 50, 100 dan 200 μg/mL. Uji viabilitas dengan WST-1 menunjukkan bahwa variasi konsentrasi eucalyptol tersebut belum dapat menekan viabilitas sel HeLa (p>0,05 dengan uji ANOVA). Selanjutnya, deteksi apoptosis (menggunakan pewarna Annexin V-FITC) dengan mikroskop fluoresens menunjukkan bahwa variasi konsentrasi eucalyptol dapat menginduksi apoptosis dibandingkan nekrosis. Meskipun demikian, belum dilakukan pengamatan pada sel hidup sehingga tidak diketahui persentase sel apoptosis dan nekrosis dalam populasi total. Pengujian lanjutan dengan pewarna DAPI yang mewarnai sel hidup dan metode kuantifikasi lainnya untuk validasi hasil pengamatan dengan mikroskop fluoresens diperlukan. ......Cervical cancer is one type of cancer with a high case prevalence rate in Indonesia. Based on GLOBOCAN 2020, found 36,633 (36% of total cancer cases in women) new cervical cancer cases and 21,003 deaths. Treatment of cervical cancer is currently still causing side effects such as drug resistance in a number of cases so that alternative treatment is needed. One of the compounds that have anticancer potential is Eucalyptol. Eucalyptol is the main compound in eucalyptus plants such as Eucalyptus globulus. The study of the effect of eucalyptol concentration on the viability and detection of apoptosis of HeLa cells was carried out at concentrations of 25, 50, 100 and 200 g/mL. The viability test with WST-1 showed that the variation in eucalyptol concentration had not been able to suppress HeLa cell viability (p>0.05 by ANOVA test). Furthermore, detection of apoptosis (using Annexin V-FITC dye) by fluorescent microscopy showed that variations in eucalyptol concentration could induce apoptosis rather than necrosis. However, no observations have been made on living cells so that the percentage of apoptotic and necrotic cells in the total population is unknown. Further testing with DAPI dye that stains living cells and other quantification methods for validation of observations by fluorescent microscopy is required.

Abstrak :
ABSTRAK
Latar belakang. Karsinoma medular sulit dibedakan secara histopatologik dan imunohistokimia dengan karsinoma invasif NST dengan gambaran medular derajat 3, karena beberapa gambaran yang tumpang tindih. Pembedaannya sangat penting terkait perbedaan tatalaksana dan prognosis. Karsinoma invasif NST dengan gambaran medular derajat 3 dianggap varian dari karsinoma invasif NST derajat 3, sehingga dapat mewakilinya. Karsinoma medular menunjukkan indeks apoptosis yang lebih tinggi dibandingkan karsinoma invasif NST derajat 3. Tujuan penelitian ini adalah mengetahui apakah indeks apoptosis dapat digunakan untuk mempertajam diagnosis karsinoma payudara medular secara obyektif menggunakan indeks apoptosis. Bahan dan Cara. Dilakukan penelitian retrospektif observasional analitik secara potong lintang terhadap 20 kasus karsinoma medular dan 20 kasus karsinoma invasif NST derajat 3. Dilakukan penilaian indeks apoptosis dengan metode TUNEL (terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate in situ nick endlabeling); selanjutnya membandingkan nilai keduanya dan menghitung titik potongnya. Dari titik potong yang didapat, selanjutnya dibandingkan indeks apoptosisnya pada sediaan simulasi core biopsy dan sediaan mastektomi/eksisinya pada kedua kasus. Hasil. Indeks apoptosis (IA) pada karsinoma medular lebih tinggi secara bermakna dibandingkan karsinoma invasif NST derajat 3 ( p 0,001). Berdasarkan kurva ROC, kami mendapatkan titik potong yang optimal pada IA 1.25. Uji kappa terhadap keselarasan sediaan core biopsy dan eksisi/mastektomi mendapatkan hasil 0,3. Kesimpulan. IA dapat digunakan untuk mempertajam diagnosis karsinoma meduler payudara pada sediaan eksisi/mastektomi. Didapatkan titik potong IA: dinyatakan ´medular´ apabila lebih besar/ sama dengan 1,25. IA potensial dapat membantu pada sediaan core biopsy jika >1.25 pada gambaran histopatologik yang memenuhi sebagian kriteria karsinoma medular.

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ABSTRACT
Background. Difficulties are often faced to differentiate between medullary breast carcinoma and invasive carcinoma of no special type with medullary features grade 3, due to morphology and immunohistochemistry overlapping features. It is important to differentiate between them due to differences in the treatment and prognosis . Invasive carcinoma NST with medullary features grade 3 is considered a variant of invasive carcinoma NST grade 3 so it can represent it. Some study showed that apoptotic index in medullary breast carcinoma is higher than invasive carcinoma of no special type grade 3. The aim of this study is to investigate whether apoptotic index can be more definitive in diagnosing medullary breast carcinoma. Patients and methods. This is a retrospective-analytic cross-sectional study using 20 cases of medullary breast carcinoma and 20 cases of invasive carcinoma of no special type grade 3. Apoptotic cell were assessed by TUNEL and the apoptotic index (AI) was calculated. Results. AI in medullary breast carcinoma is significantly higher than invasive carcinoma of no special type grade 3 (p 0,001). The cut off point of AI between medullary carcinoma and invasive carcinoma NST grade 3 is 1.25. Kappa test was done to determine the concordance between core biopsy simulation AI with the related excision/mastectomy and the result is 0,3. Conclusion. The AI can be used to improve diagnostic accuracy of medullary breast carcinoma in excision/mastectomy. The cut off point of the apoptotic index between medullary carcinoma and invasive carcinoma NST grade 3 is 1.25. Only if AI >1.25 can potentially be used to support the diagnosis of medullary carcinoma in core biopsy in case showing some of the medullary carcinoma morphologic criteria.

Abstrak :
Apoptosis or programmed cell death is a normal condition for development and live multicellular organism. Apoptosis is a morphological phenomenon that play important role in physiological processes during fetal development and in adult. Mitochondria play an important role in apoptosis. Mitochondria can do apoptosis directly. Mitochondria has 2 family of protein Bcl-2. Bcl-2 and Bcl-XL are anti apoptosis while Bad and Bax are pro apoptosis. There are 3 different mechanism apoptosis. One generated by signals arising within the cell another triggered by death activators binding to receptors at the cell surface and a third may be triggered by dangerous agent that different from two ways before. Apoptosis also need caspase as cell death executor. Study of apoptosis still done especially in case of disease. Some disease have known related with disturbing of apoptosis mechanism for example cancer and auto immun. This article reviews about molecular mechanism of apoptosis for understanding disease and future therapy.

Abstrak :
Latar belakang: Buceng {kombinasi pasak bumi (Eurycoma longifolia Jack) dan purwoceng (Pimpinella alpine Molk)}telah terbukti meningkatkan kadar testosteron (Te) dan menurunkan apoptosis. Namun belum ada bukti apakah efek tersebut dimediasi oleh penurunan ekspresi caspase3. Tujuan penelitian ini adalah untuk mempelajari apakah pemberian buceng dapat menurunkan ekspresi caspase3 sel penis dan prostat pada tikus jantan Sprague Dawley. Metode: Studi eksperimental dilakukan pada 24 tikus jantan galur Sprague Dawley, umur 90 hari dengan berat badan (BB) + 300 g, dibagi menjadi 4 kelompok secara acak masing-masing terdiri dari 6 ekor. Kelompok A, tikus dikastrasi dan diberi buceng 50 mg. Kelompok B, tikus tanpa dikastrasi, langsung dimatikan sebagai kontrol positif. Kelompok C, tikus dikastrasi dan diberi akuades 2 mL, sebagai kontrol negatif. Kelompok D, tikus dikastrasi dan diberi mesterolone 6,75 mg yang dilarutkan dalam air. Analisis statistik yang digunakan untuk menguji perbedaan ekspresi caspase3 adalah uji MANOVA, dilanjutkan dengan Post Hoc. Hasil: Analisis MANOVA pada empat kelompok menunjukkan perbedaan ekspresi caspase3 yang bermakna (p = 0,000). Analisis tes Post Hoc menunjukkan bahwa ekspresi caspase3 penis dan prostat pada kelompok A (buceng) (33,56; 35,83) lebih rendah bermakna dibanding kelompok C (kontrol negatif) (54,33;60,07) dan kelompok D (mesterolone) (51,91;56,21), p = 0,000, dan lebih tinggi dibanding kelompok B (kontrol positif atau tikus normal) (29,40; 27,72), namun secara statistik tidak bermakna ( p = 0,826). Kesimpulan: Pemberian buceng 50 mg/hari selama 30 hari berturut-turut dapat menurunkan ekspresi caspase3 pada sel penis dan prostat.

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Abstract
Background: Buceng {combination of pasak bumi (Eurycoma longifolia Jack) and purwoceng (Pimpinella alpine Molk)}has been proven to increase testosterone (Te) level and decrease apoptosis. Unfortunately, there is no evidence whether these effects are mediated by the declining of caspase3. Objective of this study was to evaluate whether buceng could decrease the expression of caspase3 of penis and prostate cells in Sprague Dawley male rats. Methods: Twenty four Sprague Dawley male rats weighing 300 g (90 days old) were randomly assigned into 4 groups of 6 male rats. Group A, rats were castrated and received buceng 50 mg. Group B, rats were not castrated, sacrifices as positive control. Group C, rats were castrated and given 2 mL aquadest as negative control. Group D, rats were castrated and got of 6.75 mg mesterolone, dissolved in 2 mL water. MANOVA statistical analysis was adopted to examine the difference expression of caspase3 in all groups. The comparison of caspase3 expression between two groups exhibiting difference values were evaluated by Post Hoc test. Results: MANOVA revealed statistically significant differences in the expression of caspase3 of penis and prostate tissues among the four groups. Post Hoct test also indicated that expression of caspase3 in group A (buceng) (33.56; 35.83) was significantly lower compared to group C (negative control) (54.33; 60.07) and group D (mesterolone) (51.91;56.21), p = 0.000, and higher compared than group B or normal rats (29.40; 27.72), but statistically not significant (p = 0.826). Conclusion: The treatment of 50 mg buceng/day for 30 consecutive days could decrease caspase3 expression in penis and prostate cells.

Abstrak :
Latar belakang : Glioblastoma Multiforme (GBM) merupakan kanker otak primer yang paling umum terjadi pada orang dewasa dan merupakan glioma yang paling agresif yang diklasifikasikan oleh WHO sebagai tingkat keempat dari astrositoma. Penatalaksanaan utama GBM dengan operasi disertai dengan kemoradiasi memberikan rata-rata harapan hidup sekitar 14,6 bulan saja, ini disebabkan pertumbuhan tumor yang difus disertai tingginya resistensi. Apoptosis, dinamakan juga “kematian sel terprogram” merupakan mekanisme kematian sel yang berperan penting dalam terapi kanker dan menjadi target utama dalam berbagai tehnik terapi kanker. Dua jalur utama apoptosis adalah jalur intrinsik yang melibatkan mitokondria dan jalur ekstrinsik yang diinduksi oleh ikatan ligan dengan reseptornya. Rotenon, inhibitor rantai respirasi kompleks I mitokondria dapat menyebabkan peningkatan kadar superoksida (ROS) endogen sehingga menginduksi jalur apoptosis intrinsik melalui lepasnya sitokrom C dan agen-agen proapoptotik ke sitosol. Rotenon bersifat lipofilik dan dapat menembus sawar darah otak dengan mudah, menjadikannya kandidat yang menarik untuk digunakan pada terapi GBM. Hingga kini mekanisme apoptosis yang disebabkan oleh rotenon pada GBM masih belum jelas diketahui, karena itu penelitian ini bertujuan untuk menelusuri jalur-jalur apoptosis yang timbul akibat induksi rotenon pada galur sel Glioblastoma Multiforme T98G. Metode : Penelitian eksperimental in vitro menggunakan kultur galur sel Glioblastoma Multiforme T98G yang diinduksi stres oksidatif dengan rotenon dosis 10μM, 20μM, dan 40μM selama enam jam, selanjutnya dilakukan identifikasi morfologi sel menggunakan inverted mikroskop. Identifikasi jalur apoptosis intrinsik dengan cara menganalisis ekspresi protein sitokrom-C dan protein kaspase-9 menggunakan metode sandwich ELISA (Enzyme Linked Immuno Assay) serta identifikasi jalur apoptosis ekstrinsik dengan menganalisis ekspresi mRNA TRAIL (Tumor Necrosis Factor Related Apoptosis Inducing Ligand) menggunakan q-RTPCR (quantitative-reverse transcriptase-Polymerase Chain Reaction) selain itu dilakukan pula elektroforesis hasil amplifikasi cDNA TRAIL pada agarosa 2%. Analisis statistik dilakukan menggunakan uji Mann Whitney pada tingkat kepercayaan 95%. Hasil : Induksi rotenon 10 μM pada galur sel T98G menyebabkan meningkatnya kadar sitokrom C yang tidak disertai meningkatnya kadar kaspase-9 dan ekspresi mRNA TRAIL. Kadar sitokrom C turun menjadi setara dengan kadar pada sel kontrol saat induksi rotenon 20μM disertai meningkatnya kadar kaspase-9 dan ekspresi mRNA TRAIL yang bermakna. Sedangkan Induksi rotenon 40μM menyebabkan turunnya kadar sitokrom C, kaspase-9 dan mRNA TRAIL. Pada pemeriksaan elektroforesis hasil RT-PCR kami mendapatkan varian TRAIL sepanjang sekitar 300 bp yang ikut teramplifikasi bersama varian sTRAIL (soluble TRAIL) sepanjang 232 bp. Varian TRAIL panjang ini nampak ditranskripsi lebih banyak saat dilakukan induksi rotenon hingga 20μM sementara varian pendek transkripsinya nampak semakin berkurang. Kesimpulan : Pada penelitian kami, induksi rotenon 20μM dapat menyebabkan terinduksinya jalur apoptosis intrinsik yang melibatkan kaspase serta jalur apoptosis ekstrinsik melalui pengaturan post transkripsi mRNA TRAIL pada galur sel Glioblastoma Multiforme T98G. ......Background : Glioblastoma Multiforme (GBM) is the most common primary brain tumor in adults and the most aggressive gliomas classified by WHO as grade IV astrocytoma. Primary treatment of GBM is surgery followed by Chemoradiation give the median survival only for about 14,6 months, this is due to difusse tumor growth with high therapy resistance. Apoptosis, named as “programmed cell death” is a mechanism of cell death that plays an important role in the treatment of cancer and being a primary target of many cancer treatment strategies. There are two central pathways of apoptosis, the intrinsic pathway that involves the mitochondria and the extrinsic pathway induced by ligands binding to the death receptors. Rotenone, respiratory chain inhibitor complex I mitochondria may increase endogenous superoxide (ROS) levels that induce intrinsic apoptotic pathway by release of cytochrome-C and several proapoptotic agents to cytosol. Rotenone is a lipophilic compound and could easily cross the Blood Brain Barrier that makes rotenone become an interesting candidate for GBM therapy. Up to now, the apoptosis mechanism induced by rotenone in GBM is not well known yet, therefore we aim to investigate the apoptosis pathways in Glioblastoma Multiforme T98G cell line induced by rotenone. Methods : This experimental study in vitro using GBM T98G cell line cultured in complete DMEM medium. We induced oxidative stress for six hours with 10 μM, 20 μM and 40 μM of rotenone respectively. After harvested, the cell morphology was identified using inverted microscope. We identified the intrinsic apoptotic pathway by analyzing the expression of cytochrome C protein and Caspase-9 protein using sandwich ELISA method, furthermore we identified the extrinsic apoptotic pathway by analyzing the expression of mRNA TRAIL using q-RT-PCR followed by gel electrophoresis to confirm the amplification of cDNA TRAIL. Statistical analysis was performed by Mann Whitney test with 95% confidence intervals. Results : Rotenone treatment of T98G resulted in increase of cytochrome C by 10μM of rotenone but no increase of caspase-9 and mRNA TRAIL. While rotenone 20 μM showed relative decrease of cytochrome C and increase of caspase-9 expression together with significant increase of the mRNA TRAIL expression (p<0,05) and induction with 40μM showed decrease of cytochrome C, caspase-9 and mRNA TRAIL expression. In the electrophoresis examination of the RT-PCR product we obtain an isoform of TRAIL (length about 300 bp) was coamplified along with our isoform (soluble TRAIL, length about 232 bp). This long isoform band become more dens in samples induced by rotenone 20μM, while the short isoform was gradually missing. Conclusions : In our study, 20μM of rotenone was able to induced both the intrinsic apoptotic pathway by caspase dependent mechanism and the extrinsic apoptotic pathway through post transcriptional regulating of mRNA TRAIL in Glioblastoma Multiforme T98G cell line.

Abstrak :
[ABSTRAK
Latar Belakang: Kanker prostat adalah kanker yang paling umum pada pria. Kanker terjadi karena hilangnya kontrol atas proliferasi sel dan apoptosis sehingga sel berproliferasi terus menerus tanpa ada kematian sel. Apoptosis diregulasi oleh beberapa protein tertentu diantaranya protein keluarga Bcl-2 dan protein kanal. Perkembangan kanker prostat memerlukan transformasi dari sel epitel yang normal menjadi sel ganas yang kehilangan kemampuan untuk mengakumulasi zinc. Salah satu efek utama zinc adalah mencegah pertumbuhan sel kanker prostat dengan menginduksi apoptosis dengan memfasilitasi proses pembentukan pori Bax yang memulai apoptogenesis mitokondria. Selain keluarga Bcl-2, VDAC1 juga berperan penting dalam proses apoptosis. Beberapa penelitian menyatakan Bcl-2 mempunyai kaitan erat dengan VDAC1 terkait proses apoptosis dan protein pro-apoptotik Bax juga secara langsung berinteraksi dengan VDAC yang kemudian menginduksi keluarnya sitokrom c dari membran mitokondria. Tujuan: Mengevaluasi ekspresi mRNA dari gen mengkode keluarga protein Bcl-2 (Bax dan Bcl-2) dalam proses apoptogenesis pada galur sel kanker prostat yg diinduksi oleh zinc; Mengevaluasi ekspresi mRNA dari gen VDAC1 dalam proses apoptogenesis pada galur sel kanker prostat yang diinduksi oleh zinc; Menganalisis hubungan antara ekspresi VDAC1 dengan protein keluarga Bcl-2 pada apoptogenesis galur sel kanker prostat. Desain: Penelitian ini menggunakan eksperimental in vitro dan analisis statistik Metode: Untuk memperbanyak galur sel kanker prostat (PC3) dilakukan kultur sel, kemudian diberi perlakuan dengan tiga kelompok (kontrol, zinc 20 μM dan staurosporin 0,16 μM). Selanjutnya dilakukan isolasi RNA dan elektroforesis RNA untuk mengetahui keutuhan RNA. Terakhir dilakukan qRT PCR yang kemudian datanya dianalisis secara statistika. Hasil: Ekspresi Bax, Bcl-2 dan VDAC1 pada galur sel kanker prostat (PC-3) yang diberi perlakuan zinc mengalami penurunan dibandingkan dengan kontrol (tidak diberi perlakuan). Akan tetapi penurunan ekspresi tersebut tidak bernilai signifikan karena nilai p > 0,05 (nilai signifikansi Bax = 0,309; nilai signifikansi Bcl-2 = 0,236; nilai signifikansi VDAC1 = 0,437). VDAC1 mempunyai korelasi yang signifikan (p < 0,05) dengan Bax (p = 0,01) dibandingkan dengan Bcl-2 (p = 0,118). Kesimpulan: Terjadi perubahan ekspresi pada setiap gen (Bax, Bcl-2 dan VDAC1) pada galur sel kanker prostat yang diberi perlakuan zinc dengan yang tidak diberi perlakuan, akan tetapi tidak bernilai signifikan. VDAC1 mempunyai korelasi yang bermakna dengan Bax dan mempunyai korelasi yang tidak bermakna dengan Bcl-2. ABSTRACT
Background: Prostate cancer is the most common cancer in men. Cancer occurs due to loss control of cell proliferation and apoptosis thus continuously proliferating cells without cell death. Apoptosis is regulated by specific proteins including Bcl-2 family proteins and channel proteins. The development of prostate cancer requires the transformation of normal epithelial cells into malignant cells that lose the ability to accumulate zinc. One of the main effects of zinc is to prevent the growth of prostate cancer cells by inducing apoptosis by facilitating the process of pore formation Bax that started apoptogenesis mitochondrial. In addition to Bcl-2 family, VDAC1 also plays an important role in the process of apoptosis. Some studies suggest Bcl-2 has close links with related VDAC1 apoptosis and pro-apoptotic protein Bax also directly interact with VDAC which then induces the release of cytochrome c from the mitochondrial membrane. Objective: To evaluate the expression of mRNA of the gene encoding the Bcl-2 family proteins (Bax and Bcl-2) in the process apoptogenesis on prostate cancer cell line that is induced by zinc; Evaluate the mRNA expression of genes in the process VDAC1 apoptogenesis on prostate cancer cell line induced by zinc; Analyzing the relationship between the expression of VDAC1 with Bcl-2 family proteins in prostate cancer cell lines apoptogenesis. Design: This study used an experimental in vitro and statistical analysis Methods: To reproduce the prostate cancer cell lines (PC3) performed cell culture, then treated with three groups (control, zinc 20 μM and staurosporin 0,16 μM). Furthermore, the isolation of RNA and RNA electrophoresis to determine the integrity of the RNA. Recently performed qRT PCR and the data were analyzed statistically. Results: The expression of Bax, Bcl-2 and VDAC1 on prostate cancer cell line (PC-3) were treated with zinc decreased than the control (untreated). However, a decrease in the expression of no significant value because the value of p > 0.05 (Bax significant value = 0.309; the value of the significance of Bcl-2 = 0.236; VDAC1 significant value = 0.437). VDAC1 has a significant correlation (p < 0.05) with Bax (p = 0.01) than Bcl-2 (p = 0.118). Conclusion: There is a change in the expression of each gene (Bax, Bcl-2 and VDAC1) in prostate cancer cell lines that treated with zinc than untreated, but no significant value. VDAC1 has a significant correlation with Bax and had no significant correlation with Bcl-2.;Background: Prostate cancer is the most common cancer in men. Cancer occurs due to loss control of cell proliferation and apoptosis thus continuously proliferating cells without cell death. Apoptosis is regulated by specific proteins including Bcl-2 family proteins and channel proteins. The development of prostate cancer requires the transformation of normal epithelial cells into malignant cells that lose the ability to accumulate zinc. One of the main effects of zinc is to prevent the growth of prostate cancer cells by inducing apoptosis by facilitating the process of pore formation Bax that started apoptogenesis mitochondrial. In addition to Bcl-2 family, VDAC1 also plays an important role in the process of apoptosis. Some studies suggest Bcl-2 has close links with related VDAC1 apoptosis and pro-apoptotic protein Bax also directly interact with VDAC which then induces the release of cytochrome c from the mitochondrial membrane. Objective: To evaluate the expression of mRNA of the gene encoding the Bcl-2 family proteins (Bax and Bcl-2) in the process apoptogenesis on prostate cancer cell line that is induced by zinc; Evaluate the mRNA expression of genes in the process VDAC1 apoptogenesis on prostate cancer cell line induced by zinc; Analyzing the relationship between the expression of VDAC1 with Bcl-2 family proteins in prostate cancer cell lines apoptogenesis. Design: This study used an experimental in vitro and statistical analysis Methods: To reproduce the prostate cancer cell lines (PC3) performed cell culture, then treated with three groups (control, zinc 20 μM and staurosporin 0,16 μM). Furthermore, the isolation of RNA and RNA electrophoresis to determine the integrity of the RNA. Recently performed qRT PCR and the data were analyzed statistically. Results: The expression of Bax, Bcl-2 and VDAC1 on prostate cancer cell line (PC-3) were treated with zinc decreased than the control (untreated). However, a decrease in the expression of no significant value because the value of p > 0.05 (Bax significant value = 0.309; the value of the significance of Bcl-2 = 0.236; VDAC1 significant value = 0.437). VDAC1 has a significant correlation (p < 0.05) with Bax (p = 0.01) than Bcl-2 (p = 0.118). Conclusion: There is a change in the expression of each gene (Bax, Bcl-2 and VDAC1) in prostate cancer cell lines that treated with zinc than untreated, but no significant value. VDAC1 has a significant correlation with Bax and had no significant correlation with Bcl-2., Background: Prostate cancer is the most common cancer in men. Cancer occurs due to loss control of cell proliferation and apoptosis thus continuously proliferating cells without cell death. Apoptosis is regulated by specific proteins including Bcl-2 family proteins and channel proteins. The development of prostate cancer requires the transformation of normal epithelial cells into malignant cells that lose the ability to accumulate zinc. One of the main effects of zinc is to prevent the growth of prostate cancer cells by inducing apoptosis by facilitating the process of pore formation Bax that started apoptogenesis mitochondrial. In addition to Bcl-2 family, VDAC1 also plays an important role in the process of apoptosis. Some studies suggest Bcl-2 has close links with related VDAC1 apoptosis and pro-apoptotic protein Bax also directly interact with VDAC which then induces the release of cytochrome c from the mitochondrial membrane. Objective: To evaluate the expression of mRNA of the gene encoding the Bcl-2 family proteins (Bax and Bcl-2) in the process apoptogenesis on prostate cancer cell line that is induced by zinc; Evaluate the mRNA expression of genes in the process VDAC1 apoptogenesis on prostate cancer cell line induced by zinc; Analyzing the relationship between the expression of VDAC1 with Bcl-2 family proteins in prostate cancer cell lines apoptogenesis. Design: This study used an experimental in vitro and statistical analysis Methods: To reproduce the prostate cancer cell lines (PC3) performed cell culture, then treated with three groups (control, zinc 20 μM and staurosporin 0,16 μM). Furthermore, the isolation of RNA and RNA electrophoresis to determine the integrity of the RNA. Recently performed qRT PCR and the data were analyzed statistically. Results: The expression of Bax, Bcl-2 and VDAC1 on prostate cancer cell line (PC-3) were treated with zinc decreased than the control (untreated). However, a decrease in the expression of no significant value because the value of p > 0.05 (Bax significant value = 0.309; the value of the significance of Bcl-2 = 0.236; VDAC1 significant value = 0.437). VDAC1 has a significant correlation (p < 0.05) with Bax (p = 0.01) than Bcl-2 (p = 0.118). Conclusion: There is a change in the expression of each gene (Bax, Bcl-2 and VDAC1) in prostate cancer cell lines that treated with zinc than untreated, but no significant value. VDAC1 has a significant correlation with Bax and had no significant correlation with Bcl-2.]

Abstrak :
ABSTRAK
Sepsis merupakan penyakit umum di perawatan intensif dan hampir 1/3 pasien yang dirawat di ICU adalah pasien sepsis. Banyak penelitian dilakukan untuk mencari penanda sepsis yang handal dan jumlah apoptosis limfosit mulai banyak diteliti sebagai penanda sepsis. Apoptosis limfosit terjadi mulai 24 jam pertama setelah onset sepsis. Saat ini belum terdapat data yang menunjukkan dapat digunakannya jumlah apoptosis limfosit sebagai penanda prognostik sepsis. Tujuan penelitian ini adalah mengetahui dapat tidaknya jumlah apoptosis limfosit digunakan sebagai penanda prognostik pada pasien sepsis berat

Desain penelitian adalah uji prognosis secara prospektif, terdiri dari 30 pasien sepsis berat dibagi berdasarkan mortalitas 14 hari, yaitu 15 pasien hidup dan 15 pasien meninggal. Diagnosis sepsis berdasarkan modifikasi definisi sepsis oleh International Sepsis Definitions Conference 2001. Jumlah apoptosis limfosit dihitung menggunakan metode flowcytometry dengan reagen antibodi monoklonal CD45 berlabel PerCP, Annexin V berlabel FITC, dan Propidium Iodide. Pada kedua kelompok tersebut dicatat data karakteristik subyek dan dilakukan penghitungan jumlah apoptosis limfosit

Rerata jumlah apoptosis limfosit pada kelompok pasien hidup adalah 0,992% dengan simpang baku 0,44% dan rerata jumlah apoptosis limfosit pada kelompok pasien meninggal adalah 1,5853% dengan simpang baku 0,57%. Jumlah apoptosis limfosit pada kedua kelompok berbeda bermakna dengan nilai p 0,004. Ditentukan nilai cut-off jumlah apoptosis limfosit 0,97%untuk menentukan prognosis pasien sepsis, dengan AUC 0,791 (IK 95% 0,631 ? 0,951), sensitivitas 86,7%, dan spesifisitas 60%. Kurva Kapplan Meier berdasarkan nilai cut-off 0,97% menunjukkan gambar yang memenuhi asumsi proporsional hazard dengan rasio hazard 0,182 (IK 95% 0,041 - 0,814), p = 0,026. Kami menyimpulkan jumlah apoptosis limfosit pasien sepsis berat dapat digunakan untuk memprediksi pasien yang meninggal dilihat dari mortalitas 14 hari, dengan nilai AUC sedang. Cut-off jumlah apoptosis limfosit 0,97% dapat digunakan sebagai cut-off dalam tatalaksana pasien sepsis berat

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ABSTRACT
Sepsis is a common illness in intensive care, almost 1/3 of patients admitted to the ICU were sepsis patients. There are plenty of researches to find a reliable marker of sepsis and the number of apoptotic lymphocytes began widely studied as a marker of sepsis. Apoptosis of lymphocytes occurred from the first 24 hours after the onset sepsis. There are currently no data on whether the number of apoptotic lymphocytes can be used as a prognostic marker of sepsis. The purpose of this study was to determine whether the number of apoptotic lymphocytes can be used as a prognostic marker in patients with severe sepsis

This was a prospective prognosis study, consisting of 30 severe sepsis patients grouped based on 14-day mortality, 15 patients are survivors and 15 patients are nonsurvivors. The diagnosis of sepsis is based on a modified definition of sepsis by the International Sepsis Definitions Conference 2001. The number of apoptotic lymphocytes was calculated using flowcytometry with PerCP-labeled anti-CD45 monoclonal antibody, FITC-labeled Annexin V, and Propidium Iodide. In both groups, characteristics of subjects were recorded and the number of apoptotic lymphocytes was calculated.

The mean of apoptotic lymphocytes in the survivor group is 0.992% with a standard deviation of 0.44%, and the mean of apoptotic lymphocytes in the nonsurvivor group is 1.5853% with a standard deviation of 0.57%. The difference between the two groups is significant with p = 0.004. This study yields an apoptotic lymphocytes cut-off value of 0.97% to determine prognosis of severe sepsis patients, with AUC of 0.791 (CI 95% from 0.631 to 0.951), 86.7% sensitivity and 60% specificity. Kapplan Meier curve based on the 0.97% cut-off demonstrates that hazard proportion is fulfilled with hazard ratio of 0.182 (95% CI 0.041 to 0.814) and p= 0.026. It is concluded that the number of apoptotic lymphocytes in severe sepsis patients can be used to predict nonsurvivors based on 14-day mortality, with moderate AUC. The apoptotic lymphocytes cut-off value of 0.97% can be used as a cut-off for severe sepsis patient management