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"ABSTRAKIn vivo apoptosis of fibroblast pulp cells by ionizing radiation from radiotherapy of the head and neck area has not yet been demonstrated. The study aimed to show in vivo the effect of a single dose of ionizing radiation on apoptosis of fibroblast pulp cells. The sample group consisted of 24 healthy male Wistar rats that were 3-4 months old and 150- 200 g in weight. The rats were divided into 4 groups of 6 rats that were subjected to Cobalt 60 radiation to the head at the levels of 0, 100, 200 or 400 rad. The rats were sacrificed 24 hours after radiation exposure, and the lower incivus were taken for histopatological processing. Apoptosis was detected by using the TUNEL Assay method. The apoptotic fibroblast pulp cells were counted under light microscope by multiple observers using the blind test approach. The fraction of apoptotic cells was counted as mean of labial and palatal sides of the teeth below odontogenic and free-cell zone. The data were statistically analyzed using one-way anova. The results showed the percentage of apoptotic of fibroblast pulp cells was 6.4, 23.7, 34.5 and 17.8% after 0, 100, 200 and 400 rad doses, respectively. There were significant differences in the apoptotic percentages between the four groups (p<0.05). In conclusion, the highest fraction of apoptotic fibroblast pulp cells was found after a single 200 rad dose, and this fraction decreased after a single dose of 400 rad."

"ABSTRAKLatar Belakang: Vitrifikasi merupakan suatu teknik untuk menjaga sel dari kerusakan saat proses simpan beku tanpa adanya pembentukan kristal es. Keberhasilan vitrifikasi ditentukan oleh beberapa faktor diantaranya: jenis dan konsentrasi krioprotektan. Telah banyak penelitian vitrifikasi oosit dengan menggunakan berbagai macam jenis krioprotektan, namun belum diperoleh hasil yang optimal.Tujuan: untuk mengetahui efek sukrosa dan trehalosa pada medium kriopreservasi terhadap morfologi, permeabilitas membran mitokondria, dan apoptosis oosit mencit strain DDY setelah simpan bekuMetode: Mencit Mus musculus albinus betina strain DDY usia 8 minggu disuperovulasi dengan 10 IU Gonadotropin dan diinduksi dengan10 IU hCG. Lima belas jam kemudian, oosit dikoleksi, lalu divitrifikasi dengan menggunakan Equilibrium Solution ES yaitu 7,5 DMSO dan 7,5 Ethylene Glycol EG , dan Vitrification Solution VS yang terdiri dari: VS1 berupa 16,5 DMSO ditambah 16,5 EG ditambah 0,5 M sukrosa, sedangkan VS2 berupa 16,5 DMSO ditambah 16,5 EG ditambah 0,5 M trehalosa. Selanjutnya oosit diletakkan di dalam cryotop dan dimasukkan ke dalam nitrogen cair. Warming dilakukan dengan memasukkan oosit pada Warming Solution WS yakni: WS1a berupa 0,3 M sukrosa dan WS1b berupa 0,15 M sukrosa, sedangkan WS2a berupa 0,3 M trehalosa dan WS2b berupa 0,15 M trehalosa. Oosit yang telah di-warming lalu dianalisis morfologinya, permeabilitas membran mitokondria, dan apoptosisnya.Hasil: Pada kelompok medium sukrosa, didapatkan 85.7 oosit dengan morfologi normal, rasio intensitas pendaran merah per hijau 3.57, dan 84.6 oosit dengan TUNEL negatif. Di lain pihak, pada kelompok medium trehalosa, didapatkan 93.1 oosit dengan morfologi normal, rasio intensitas pendaran merah per hijau 3.79, dan 92.3 oosit yang TUNEL negatif.Kesimpulan: Trehalosa memiliki efek yang lebih baik pada oosit setelah simpan beku dibandingkan sukrosa

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ABSTRACTBackground Vitrification is a cryopreservation method used in assisted reproductive technology ART . Vitrification preserves cells and prevents from cryodamage by eliminating ice crystal formation. The successful of vitrification depends on type and concentration of cryoprotectant. Although there are many researches about oocyte vitrification, there are still no appropriate kind and composition of cryoprotectant which give the optimum result.Aim This research was aimed to analyze the effect of sucrose and trehalose as cryoprotectant on morphology, mitochondrial membrane potential, and apoptotic status of mice oocyte DDY strain after cryopreservationMethod DDY female mice 8 weeks old were superovulated with 10 IU Gonadotropin Gonal F followed by 10 IU Pregnil 48 hours later. Oosit were collected 15 hrs after Pregnyl injection and cumulus cell were removed. Cumulus free oocytes were vitrified in two different Vitrification Solution VS VS1 16,5 DMSO, 16,5 EG, and 0,5 M sucrose in HM, VS2 16,5 DMSO, 16,5 EG, and 0,5 M trehalose in HM using cryotop. Two steps warming was performed with Warming Solution WS WS1a 0,3 M sucrose and WS1b 0,15 M sucrose, besides WS2a 0,3 M trehalose and WS2b 0,15 M trehalose . Then, the warmed oocytes was analyzed based on morphology, mitochondrial membrane potential and their apoptotic status.Result The sucrose group showed 85.7 oocytes with normal morphology, 3.57 fluorescence red per green intensity, and 84.6 negative TUNEL oocytes. While, trehalose group showed 93.1 oocytes with normal morphology, 3.79 fluorescence red per green intensity, and 92.3 negative TUNEL oocytes.Conclusion Trehalose has better effect for oocytes in vitrification than sucrose."

"Sel kanker kanker payudara jenis TNBC memiliki kadar MnSOD yang tinggi. Ekspresi MnSOD yang tinggi pada sel kanker TNBC dapat menghasilkan sel dengan kapasitas proliferasi tinggi, apoptosis minimal, dan resistensi terapeutik. Keberadaan teknologi edit genom CRISPR/Cas9 yang menargetkan MnSOD diyakini dapat menurunkan ekspresi MnSOD sehingga mengurangi aktivitas proliferasi sel kanker BT-549 dan meningkatkan apoptosisnya. MnSOD diketahui mengandung polimorfisme yang meningkatkan risiko berkembangnya kanker. Oleh karena itu, sangat penting untuk menganalisis ekson 2 dari pengeditan genom gen MnSOD pada sel BT-549. Pada penelitian ini, memanfaatkan kultur sel BT-549 untuk diperbanyak dan dilakukan pemisahan DNA, RNA, dan protein dari sel kultur. Analisis western blot MnSOD, survivin, caspase 3, caspase 9, cleaved caspase 3, dan cleaved caspase 9. Selain itu, ekspresi relatif mRNA MnSOD dan survivin dihitung menggunakan teknik qRT-PCR. Selain itu dilakukan sekuensing DNA, pemodelan homologi protein, dan prediksi protein pengeditan genom. Analisa fenotip terdapat analisis proliferasi sel juga dilakukan untuk menentukan waktu penggandaan, serta studi siklus sel flowcytometric. Pengukuran flow cytometric dari apoptosis juga dilakukan untuk menguji pengaruh genome editing pada MnSOD. Hasil pada pengeditan genom yang difokuskan pada ekson 2 gen MnSOD dapat menurunkan ekspresi relatif mRNA MnSOD dan ekspresi protein MnSOD. Selain itu, adanya genome editing pada MnSOD juga mempengaruhi pola proliferasi klon KO 11.3 dan 11.4 yang lebih lambat jika dibandingkan dengan sel WT; hal ini sesuai dengan hasil analisis siklus sel, yang menunjukkan persentase fase G0/G1 yang tinggi dan persentase fase S, G2/M yang rendah pada klon KO 11.3 dan 11.4 jika dibandingkan dengan WT. Penghambatan ekspresi MnSOD juga dapat meningkatkan kemampuan apoptosis sel, yang ditunjukkan dengan tingginya proporsi sel yang mengalami apoptosis yang diukur dengan flow cytometry, serta peningkatan kadar protein inisiator dan efektor apoptosis. Kesimpulannya, KO gen MnSOD pada sel BT-549 mengurangi ekspresi protein MnSOD, sehingga menghambat pertumbuhan sel dan mendorong kematian sel BT-549.

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It is known that TNBC cancer cells have a high level of MnSOD. High MnSOD expression in TNBC cancer cells can result in cells with a high proliferation capacity, minimal apoptosis, and therapeutic resistance. It is believed that the existence of CRISPR/Cas9 genome editing technology that targets MnSOD can decrease MnSOD expression, hence reducing the proliferative activity of BT-549 cancer cells and increasing their apoptosis. MnSOD is known to include polymorphisms that increase the risk of developing cancer. Therefore, it is crucial to analyze exon 2 of the MnSOD gene genome editing in BT-549 cells. Methods: This work utilized BT-549 cell culture to proliferate cells and to separate DNA, RNA, and protein from the cultured cells. Western blot analysis of MnSOD, survivin, caspase 3, caspase 9, cleaved caspase 3, and cleaved caspase 9. Additionally, the relative expression of MnSOD and survivin mRNAs was calculated using the qRT-PCR technique. There was DNA sequencing, protein homology modeling, and genome editing protein prediction. In phenotypic analysis, cell proliferation analysis was also performed to determine doubling time, in addition to flowcytometric cell cycle study. Flow cytometric measurement of apoptosis was also performed to examine the effect of genome editing on MnSOD. Result: Genome editing focused at exon 2 of the MnSOD gene can reduce the relative expression of MnSOD mRNA and the expression of MnSOD protein. In addition, the existence of genome editing in MnSOD also affected the slower proliferation pattern of KO clones 11.3 and 11.4 when compared to WT cells; this is consistent with the results of cell cycle analysis, which demonstrated a high percentage of G0/G1 phase and a low percentage of S phase, G2/M, in KO clones 11.3 and 11.4 when compared to WT. The inhibition of MnSOD expression can also boost the capability of cell apoptosis, as indicated by the high proportion of cells undergoing apoptosis as measured by flow cytometry, as well as the elevated levels of apoptosis initiator and effector proteins. Conclusion: MnSOD gene knockout on BT-549 cells reduces MnSOD protein expression, hence inhibiting cell growth and promoting BT-549 cell death."

"Dengue merupakan mosquitos borne virus yang paling cepat menyebar di dunia dan menjadi masalah kesehatan utama di daerah tropis dan subtropis. Cedera dan disfungsi hati merupakan fitur penting yang terlihat pada infeksi virus dengue (DENV) pada manusia. Patogenesis kerusakan sel hati akibat infeksi DENV belum dapat dijelaskan secara pasti dan melibatkan banyak mekanisme yang berbeda, terutama mekanisme infeksi virus secara langsung dan secara tidak langsung, melalui respon imun tubuh akibat sekresi sitokin yang berlebih. Untuk mengetahui infeksi DENV pada sel hati serta peranan beberapa sitokin pada sel hati akibat infeksi DENV, maka pada penelitian ini dilakukan infeksi DENV pada sel Huh 7it-1 yang dikultur bersama PBMC dan dilakukan pengukuran kadar sitokin yang berperan dalam infeksi DENV pada sel Huh 7it-1 menggunakan ELISA. Selain itu, pada penelitian ini juga dilakukan uji infektivitas DENV menggunakan FFU assay, pengukuran viabilitas sel Huh 7it-1 menggunakan MTT assay, pengukuran kadar ALT serta AST, dan uji Flow Cytomeri untuk mengetahui apoptosis serta nekrosis sel Huh 7it-1 akibat infeksi DENV pada sel hati. Hasil menunjukkan bahwa Infeksi DENV-2 pada sel Huh 7it-1 yang dikultur bersama PBMC menyebabkan peningkatan kadar ALT dan AST, penurunan viabilitas sel yang dipicu oleh apoptosis serta nekrosis sel Huh 7it-1. Selain itu, terjadi peningkatan kadar sitokin IL-1α, IL-1β, dan IP-10, dimana peningkatan kadar IL-1α dan IL-1β diketahui berkorelasi dengan penurunan viabilitas sel, peningkatan apoptosis serta peningkatan nekrosis sel Huh 7it-1. Namun, peningkatan kadar IP-10 diketahui hanya berkorelasi dengan peningkatan kejadian apoptosis sel Huh 7it-1. Selanjutnya, peningkatan apoptosis serta nekrosis pada sel Huh 7it-1 yang terinfeksi DENV-2 baik yang dikultur bersama PBMC maupun tidak, diketahui berkorelasi dengan penurunan viabilitas sel Huh 7it-1.

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Dengue is a mosquito borne virus that spreads rapidly in the world and is a major health problem in the tropics and subtropics region. Human infections are often associated with liver injury and dysfunction. The pathogenesis of liver cell damage due to DENV infection is not clear and many different mechanisms are involve. Especially, the direct cytopathic effect of the virus and indirectly by the immune response due to excessive secretion of cytokines. To find out liver cell damage due to DENV infection and to know the cytokine profiles that play a role in liver cell damage, this study conducted a DENV infection in Huh 7it-1 cells co-cultured with PBMC and cytokine measurements using ELISA. In addition, this study also conducted a DENV infectivity test using FFU assay, measurement of Huh 7it-1 cell viability using MTT assay, measurement of ALT and AST levels, and Flow Cytomeri assay to determine apoptotic and necrotic cell due to DENV infection in Huh 7it-1 cells. The results showed that DENV-2 infection in Huh 7it-1 cells cocultured with PBMC caused an increased of ALT and AST levels, decreased in cell viability triggered by apoptotic and necrotic of Huh 7it-1 cells. Furthermore, increased levels of IL-1α, IL-1β, and IP-10 are higher when compared to control cells. Increased levels of IL-1α and IL-1β are known to correlate with decreased cell viability and an increased in apoptotic and necrotic cells, however an increased in IP-10 levels only correlates with an increased of apoptotic Huh 7it-1 cells. Moreover, increased cell apoptotic and necrotic in Huh 7it-1 cells infected with DENV-2 with or without PBMC, is known to correlate with decreased of Huh 7it- 1 cell viability."

"Prevalensi kanker payudara pada tahun 2020 menduduki peringkat pertama di dunia maupun di Indonesia. Pencarian obat antikanker menggunakan metode komputasi dinilai lebih efektif dan selektif. Asam galat dan turunannya (ester dan amida) merupakan senyawa yang memiliki aktivitas biologis seperti antikanker. Tujuan dari studi ini adalah melakukan analisis secara in-siliko dan in-vitro terhadap senyawa turunan asam galat (N-Alkil galamida) sebagai agen apoptosis sel kanker payudara MCF7. Sebelas senyawa N-Alkil galamida yang telah disintesis dilakukan studi in-siliko meliputi, interaksi protein-protein, interaksi obat-protein, analisis ADMET dan pemodelan molekuler. Tiga senyawa terbaik berdasarkan studi in-siliko diuji aktivitas sitotoksisitasnya pada sel MCF7 menggunakan metode MTT dan analisis apoptosis menggunakan annexin V-FITC/PI dengan flow cytometry. Protein target terpilih, yaitu JUN, AKT1, CASP3 dan CASP7. Senyawa N-Oktil galamida, N-Ters-Butil galamida dan N-Isoamil galamida merupakan tiga senyawa terbaik. Senyawa asam galat dan turunannya, secara analisis prediksi ADMET termasuk dalam kategori aman sebagai kandidat obat. Aktivitas sitotoksik ketiga senyawa tersebut dinyatakan dengan nilai IC50 berturut-turut adalah 205.2 ± 0,44 μM, 372.6 ± 4,09 μM, dan 441.7 ± 1,41 μM. Aktivitas apoptosis mencapai 55 hingga 56% dibandingkan dengan kontrol sel. Senyawa N-Oktil galamida, N-Ters-Butil galamida dan N-Isoamil galamida berpotensi sebagai agen apoptosis pada sel kanker payudara MCF7.

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The prevalence of breast cancer in 2020 is ranked first in the world and in Indonesia. Searching for anticancer drugs using computational methods is considered more effective and selective. Gallic acid and its derivatives (esters and amides) are compounds that have biological activities such as anticancer. In this study, N-Alkyl gallamides were analyzed in-vitro and in-silico for their potential to act as apoptotic agents for MCF7 breast cancer cells. In-silico investigations on eleven N-alkyl gallamide compounds, including protein-protein interactions, drug-protein interactions, ADMET analysis, and molecular modelling, have been conducted. The three best compounds based on in-silico studies were tested for cytotoxicity activity on MCF7 cells using the MTT assay and apoptosis analysis using annexin V-FITC/PI with flow cytometry. Selected target proteins, namely JUN, AKT1, CASP3 and CASP7. According to ADMET study, gallic acid and their derivatives are safe as therapeutic candidates. The cytotoxic activities of the three compounds were expressed by IC50 values of 205.2 ± 0.44 μM, 372.6 ± 4.09 μM, and 441.7 ± 1.41 μM, respectively. Apoptotic activity reached 55 to 56% compared to control cells. The N-Octyl gallamide, N-Ters-Butyl gallamide and N-Isoamil gallamide compounds have the potential as apoptotic agents in MCF7 cells."

"Preeklamsia merupakan salah satu penyebab kematian utama ibu dan perinatal di dunia. Inflamasi disertai tingginya indeks apoptosis di syncytiotrophoblast dan ekspresi Cyclophilin A diduga berperan pada preeklamsia. Faktor tersebut diasumsikan menyebabkan jejas jantung/pembuluh darah yang meningkatkan risiko morbiditas dan mortalitas ibu dan perinatal. Tujuan penelitan ini adalah menganalisis peran inflamasi, indeks apoptosis dan Cyclophilin A terhadap jejas jantung/pembuluh darah pada preeklamsia awitan dini, lanjut dan kehamilan normal. Sebanyak 47 wanita hamil yang terpilih dilakukan pemeriksaan hematologi, ekokardiografi dan ultrasonografi Doppler. Plasenta diperiksa secara histopatologis untuk mengukur ekspresi NF-KB dan PARP-1, indeks apoptosis berdasarkan pemeriksan TUNEL, ekspresi Cyclophilin A dan pemeriksaan ultrastruktur mikroskopik pada syncytiotrophoblast. Analisis Anova digunakan untuk mengidentifikasi perbedaan antara ketiga kelompok, sedangkan regresi linier digunakan untuk mengetahui korelasi faktor yang diduga terhadap jejas jantung/pembuluh darah menggunakan SPSS 20. Usia ibu, indeks massa tubuh (IMT), hitung trombosit, NF- KB dan indeks apoptosis lebih tinggi disertai Cyclophilin A lebih rendah pada preeklamsia awitan dini dibandingkan preeklamsia awitan lanjut dan kehamilan normal. Hitung leukosit lebih tinggi pada preeklamsia awitan lanjut dibandingkan awitan dini dan normal. Total peripheral resistance (TPR) paling tinggi pada kelompok awitan dini dibandingkan awitan lanjut dan kehamilan normal, sedangkan cardiac index (CI) tidak berbeda bermakna pada ketiga kelompok. Resistensi indeks (RI) lebih tinggi pada preeklamsia awitan dini dibandingkan awitan lanjut dan kehamilan normal. Berdasarkan analisis regresi linier multivariat, membuktikan indeks apoptosis dan Cyclophilin A memiliki hubungan dengan jejas jantung/pembuluh darah. Hal tersebut menunjukkan inflamasi, indeks apoptosis, Cyclophilin A disertai pemeriksan ekokardiografi dan ultrasonografi Doppler merupakan metode yang cepat, tepat dan noninvasif faktor risiko terhadap jejas jantung/pembuluh darah pada preeklamsia. Penelitian yang dianjurkan di masa datang adalah menilai geometri jantung dengan ekokardiografi dan volumetri plasenta dengan ultrasonografi.

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Preeclampsia is one of the leading cause of maternal and perinatal death in the world. Inflammation accompanied by a high apototic index of syncytiotrophoblast and Cyclophilin A were speculated to play a role in preeclampsia. Those response were assumed to cause cardiovascular injury which lead to the risk of maternal and perinatal morbidity and mortality in preeclampsia. The objective of the study was to investigate the role of inflammation, apoptotic index and Cyclophilin A in cardiovascular injury in early and late onset preeclampsia compared to normal pregnancy.

A total of 47 pregnant women were selected, consisting almost the same size of each group (30%) and assessed for maternal hematology, echocardiography and Doppler ultrasound. Placentae were assessed histopathologically by measuring nuclear factor kappa-light-chain-enhancer of activated B cells (NF- KB) and Poly (ADP-ribose) polymerase 1 (PARP-1) expression for inflammation marker, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay for apoptotic index and Cyclophilin A. Confirmation by transmission electron microscopy (TEM) was done. Anova analysis was used to identify the differences between the three groups while linier regression was employed to assess the correlation between factors on cardiovascular injury using SPSS 20.

Maternal age, body mass index (BMI), platelet count, NF- KB and apoptotic index, resistance index (RI) were higher supported by low Cyclophilin A in early onset preeclampsia (EOP) than in late onset preeclampsia (LOP) and normal pregnancy. Leukocyte count was higher in late onset preeeclampsia than in early and normal pregnancy. Total peripheral resistance (TPR) was highest in the EOP compared to LOP and normal pregnancy, while the cardiac index (CI) was not significantly different in all groups.

Based on multivariate linear regression analysis, the apoptotic index and Cyclophilin A correlated to cardiovascular injury. Assesing inflammation, apoptotic index, Cyclophilin A, echocardiography examination and Doppler ultrasound examination might indicated timely and non-invasive detection as an alarm entry point for cardiovascular injury in both early and late onset preeclampsia. Cardiac geometry by echocardiography and placental volumetry by Doppler ultrasound should be performed in future research."

"Kanker adalah suatu penyakit dimana sel tubuh bersifat abnormal dan tumbuh secara tidak terkontrol yang bisa meluas dan menyebar ke setiap bagian di tubuh manusia. Ditemukan ekspresi berlebih procaspase-3 pada beberapa kanker yang harus diaktivasi menjadi Caspase-3 agar dapat menginduksi apotosis pada sel. Pada penelitian ini, dilakukan penapisan virtual senyawa dari basis data tanaman herbal Indonesia sebagai aktivator Procaspase-3 menggunakan peranti lunak Autodock dan Autodock Vina. Penelitian ini divalidasi menggunakan parameter Enrichment Factor EF, Receiver Operating Characteristics ROC gunakan Autodock diperoleh sepuluh senyawa terbaik dengan energi ikatan-8,28 -9,31 kkal/mol dan menggunakan Autodock Vina diperoleh sepuluh senyawa terbaik dengan energi ikatan -8,1 -8,8 kkal/mol. Didapatkan dua senyawa yang beririsan pada kedua peranti lunak, yaitu Betulinic acid dan Maslinic acid. Betulinic acid berinteraksi dengan residu dengan Leu136A, Lys137A, Tyr195A, dan Pro201 pada Autodock dan Autodock Vina. Sedangkan Maslinic acid berinteraksi pada residu Leu136A, Lys137A, dan Pro201 pada Autodock dan Autodock Vina.

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Cancer is a disease where body cell being abnormal and grow uncontrolled which can spread to every part of human body. Previous research found excessive expression of Procaspase 3 on cancer that must be activated to Caspase 3 in order to induce apoptotic in cells. In this research, virtual screening of Indonesian Herbal Database as Procaspase 3 activator was performed using Autodock and Autodock Vina software. This research was validated using Enrichment Factor EF , Receiver Operating Characteristics ROC, and Area Under Curve AUC parameters. Results of virtual screening using Autodock obtained the best ten compounds with binding energy 8,28 9,31 kcal mol and Autodock Vina obtained the best ten compounds with binding energy 8,1 8,8 kcal mol. Both virtual screening software showed two compounds in common, which is Betulinic Acid, and Maslinic acid. Betulinic acid interacts with residue Leu136A, Lys137A, Tyr195A, and Pro201 in Autodock and Autodock Vina. While Maslinic acid interacts with residue Leu136A, Lys137A, and Pro201 in Autodock and Autodock Vina."