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"Legionella. Oleh karena itu, dalam penelitian ini dikembangkan uji PCR duplex (dPCR) untuk deteksi Legionella sp. danL. peneumophila secara simultan pada sampel air tower. Metode kultur digunakan sebagai baku emas.Metode: Dilakukan optimasi metode dPCR untuk mendapatkan teknik uji yang memiliki sensitivitas dan spesifi sitastinggi. Metode kemudian diuji pada 9 sampel air tower yang diperoleh dari 9 gedung di Jakarta. Untuk metode kultur,bakteri ditumbuhkan pada media selektif ?growth factor supplemented-buffered charcoal yeast extract? (BCYE).Hasil: Dari 9 sampel yang diuji dengan dPCR, 6 menunjukkan positif Legionella sp., 1 positif L. pneumophila, dan 2menunjukkan hasil uji negatif. Untuk sampel yang sama, metode kultur menunjukkan hasil uji negatif.Kesimpulan: Uji dPCR adalah uji yang sangat sensitif dibandingkan dengan metode kultur, dan uji dPCR ini dapatdigunakan untuk pemeriksaan rutin Legionella sp. dan L. pneumophila pada sampel air dari ?tower?.

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AbstractAim: Since culture method is time-consuming and has low sensitivity, we developed a duplex PCR (dPCR) assay for thedetection of Legionella sp. and L. pneumophila in cooling tower samples. We used culture method as a gold standard.Methods: Optimization of dPCR method was performed to obtain an assay with high sensitivity and specifi city. Theoptimized method was used to detect Legionella sp. dan L. pneumophila in 9 samples obtained from 9 buildings inJakarta. For culture method, the bacteria were grown or isolated on selective growth factor supplemented-bufferedcharcoal yeast extract (BCYE) media.Results: Of 9 samples tested by dPCR assay, 6 were positive for Legionella species,1 was positive for L. pneumophila,and 2 showed negative results. For the same samples, no Legionella sp. was detected by the culture method.Conclusion: dPCR assay was much more sensitive than the culture method and was potentially used as a rapid,specifi c and sensitive test for routine detection of Legionella sp. dan for L. pneumophila in water samples."

"Metilasi DNA merupakan salah satu penyebab umum inaktivasi Mismatch Repair Gene (MMR). Gen MMR memperbaiki kesalahan penyisipan/penghapusan basa nukleotida pada proses sintesis DNA. Metilasi pada promoter gen MMR memiliki asosiasi dengan pembentukan kanker kolon, sehingga metilasi tersebut perlu diidentifikasi. Identifikasi gen MMR dapat dilakukan menggunakan teknik methylation-specific multiplex ligation-dependent probe amplification amplification (MS-MLPA). Prinsip dari teknik MS-MLPA yaitu amplifikasi probe yang menempel pada sekuens termetilasi. Tujuan dari penelitian ini yaitu untuk mengoptimasi teknik MS-MLPA dan mengidentifikasi metilasi gen MMR pada kanker kolon dengan teknik MS-MLPA. Penelitian ini menggunakan 27 sampel jaringan frozen kanker kolon yang telah tersedia di Biobank Rumah Sakit Kanker Dharmais (RSKD). Sampel tersebut dianalisis menggunakan probemix Mismatch Repair Gene [ME011-C1][C1-0518] yang telah didesain khusus untuk mendeteksi pada beberapa gen MMR yakni MLH1, PMS2, MSH6, dan MSH2. Hasil penelitian menunjukkan optimasi teknik MS-MLPA telah berhasil dilakukan, sehingga identifikasi metilasi pada gen MMR telah berhasil diperoleh pada 4 sampel pasien. Gen MMR tersebut yakni MLH1 dan MSH6, dengan persentase masing-masing 75% dan 25%.

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DNA methylation is one of the most common causes of mismatch repair gene (MMR) inactivation. The MMR gene corrects errors in the insertion/deletion of nucleotide bases in the DNA synthesis process. MMR gene promoter methylation has an association with the formation of colon cancer, so the methylation needs to be identified. Identification of the MMR gene can be done using the methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) technique. The principle of the MS-MLPA technique is the amplification of the probe attached to the methylated sequence. The purpose of this study was to optimize the MS-MLPA technique and identify MMR gene methylation in colon cancer using the MS-MLPA technique. This study used 27 samples of frozen colon cancer tissue that were available at the Dharmais Cancer Hospital Biobank (RSKD). The samples were analyzed using the Mismatch Repair Gene probemix [ME011-C1][C1-0518] which has been specially designed to detect several MMR genes, namely MLH1, PMS2, MSH6, and MSH2. The results show that the optimization of the MS-MLPA technique has been successfully carried out, so that the identification of methylation in the MMR gene has been successfully obtained in 4 patient samples. The MMR genes are MLH1 and MSH6, with a percentage of 75% and 25%, respectively. analyzed using probemix Mismatch Repair Gene [ME011-C1][C1-0518] which has been specifically designed to detect several MMR genes namely MLH1, PMS2,

MSH6, and MSH2. The results showed that the optimization of the MS-MLPA technique was successful, so that identification of the methylation in the MMR gene was successfully obtained in 4 patient samples. The MMR genes are MLH1 and MSH6, with percentages of 75% and 25% respectively."

"Cellular responses to stress including DNA damage show multiple options involving the mechanisms of growth arrest, DNA repair and programmed cell death or apoptosis. Failures in these mechanisms can result in oncogenesis or accelerated senescence. Much of the response is coordinated by p53, a nuclear phosphoprotein with a central role in the defences against physical, chemical and pathogenic agents which challenge the DNA integrity. The p53 pathways for mobilising the cellular defences are linked to the pRb/E2F pathways regulating the cell cycle progression. This paper aims to review the current understanding on the networks and main molecular machinery of these processes. In addition, the implications on cellular decision making for the defences as well as evolutionary aspects of these mechanisms are discussed in brief."

"Radiasi merupakan terapi pilihan untuk kanker serviks stadium III B, namun permasalahan timbul karena adanya sifat radioresisten. Sel punca kanker SPK merupakan salah satu faktor yang diduga berkontribusi terhadap hal tersebut. SOX2 dan OCT4 merupakan faktor transkripsi yang mengekspresikan sifat-sifat SPK, yaitu mengontrol sifat pluripoten, self-renewal, berperan pada karsinogenesis, metastasis, resistensi terhadap terapi dan rekurensi tumor. Faktor apoptosis, DNA repair dan telomerase merupakan mekanisme yang berkaitan dengan radioresisten. Penelitian ini bertujuan untuk mempelajari hubungan antara SOX2 dan OCT4 sebagai penanda SPK terhadap respons terapi radiasi, serta kaitannya dengan faktor apoptosis caspase-3 , DNA repair Chk1 dan telomerase hTERT .Penelitian ini merupakan case control, terhadap 48 kasus karsinoma sel skuamosa serviks stadium III B yang telah menjalani terapi radiasi/kemoradiasi di RS Cipto Mangunkusumo/FKUI. Kasus dibagi dalam 2 kelompok, yaitu hasil terapi komplet 27 kasus dan hasil terapi inkomplet 21 kasus . Kasus dengan respons awal terapi radiasi baik dilakukan pemeriksaan bulan Pap smear dan HPV pada bulan ke-6 atau sampai ke-12 setelah terapi. Ekspresi SOX2, OCT4, caspase-3, Chk1 dan hTERT diperiksa secara imunohistokimia dari blok parafin biopsi awal.Ekspresi kuat SOX2 dan OCT4 dengan H-score masing-masing lebih dari 96,6 dan 61,9 mempunyai hubungan bermakna dengan respons awal terapi radiasi maupun respons akhir terapi radiasi SOX2 p = 0,017, p = 0,004 dan OCT4 p < 0,001, p < 0,001 . Ditemukan hubungan bermakna antara ekspresi Chk1 dan hTERT dengan respons awal terapi radiasi Chk1 p = 0,006, hTERT p = 0,029 . Tidak ditemukan hubungan yang bermakna antara ekspresi caspase-3, Chk1, hTERT dengan ekspresi SOX2 dan OCT4. Uji multivariat menunjukkan bahwa SOX2 dan OCT4 yang paling memengaruhi respons terapi OR = 5,12, p = 0,040 dan OR = 17,03, p < 0,001, secara berurutan . Uji probabilitas menunjukkan kemungkinan respons akhir terapi radiasi inkomplet sebesar 87,91 bila ekspresi kedua penanda SPK kuat.Ekspresi kuat SOX2 dan OCT4 dapat memprediksi hasil terapi radiasi inkomplet pada karsinoma serviks stadium III B.

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Radiotherapy is the main choice of treatment for stage III B cervical cancer, but radioresistance becomes a difficult matter. Cancer stem cell is one of the factors suspected involving in radioresistant cancers. SOX2 and OCT4 are transcription factors which have pluripotent cell characteristics, and self renewal ability. They also involved in carcinogenesis, metastasis, tumor recurrent, and resistance toward therapy. Apoptotic, DNA repair, and telomerase factors are mechanisms that also contribute to radioresistance. This study aims to know the role of SOX2 and OCT4 as CSC markers, apoptotic factor caspase 3 , DNA repair Chk1 and telomerase hTERT toward radiotherapy.The design of this study was case control with 48 cases of stage III B cervical squamous cell carcinoma patients who had finished receiving radiation chemo radiation therapy at Cipto Mangunkusumo Hospital FMUI, Jakarta. They were classified in 2 groups based on the final response of treatment, which were complete and incomplete one. Pap smear and DNA HPV were performed in month 6 or until month 12 after therapy for good initial therapy. Immunohistochemistry was done to analyze SOX2, OCT4, caspase 3, Chk1 and hTERT expression from the paraffin block of initial biopsy.Strong expression of SOX2 and OCT4 with each H score was higher than 96.6, and 61.9 had significant association with both initial and final therapy response SOX2 p 0.017, p 0.004 and OCT4 p 0.001, p 0.001, repectively . There was significant association between expression of Chk1 and hTERT, and initial therapy response p 0.006 for Chk1, and p 0.029 for hTERT . No significant differences were found between caspase 3, Chk1, hTERT, and SOX2 and OCT4. Multivariate analysis showed SOX2 and OCT4 were the most influenced antibodies for radiotherapy response OR 5.12, p 0.040, and OR 17.03, p 0.001, respectively . The likelihood of incomplete final therapy response was 87.91 if the expression both of CSC markers were strong.Expression of SOX2, and OCT4 could predict the incomplete radiotherapy of stage III B cervical cancer cases. "