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"Penelitian mengenai transformasi gen cry IAc ke padi subpesies Indica, Inpari 13, IR 64, dan Inpari 6, menggunakan Agrobacterium tumefaciens telah dilakukan, namun tanaman transgenik belum berhasil diperoleh. Gen cry IAc merupakan salah satu kelompok gen cry 1 yang memiliki toksin paling tinggi terhadap hama Lepidotera khususnya hama penggerek batang. Transformasi gen cry IAc dilakukan ke kalus padi menggunakan Agrobacterium tumefaciens strain LBA4404 yang membawa plasmid pAY560325_cryIAc. Kalus padi ditumbuhkan dalam media induksi kalus N6D selama 5 hari sebelum diinfeksi dengan Agrobacterium tumefaciens strain LBA4404. Selanjutnya, kalus ditanam dalam media resting, seleksi, dan media regenerasi. Gen cry IAc dan hptII telah dikonfirmasi menggunakan teknik isolasi DNA dan PCR. Transformasi gen cry IAc berhasil dilakukan dan terdapat perbedaan respons diantara ketiga varietas terhadap perlakuan kultur jaringan dan transformasi. Inpari 13 memberikan respons terbaik terhadap perlakuan transformasi berdasarkan hasil PCR gen cry IAc dan gen hptII, sedangkan IR 64 memberikan respons terbaik terhadap perlakuan kultur jaringan berdasarkan jumlah kalus embriogenik pada media induksi kalus.

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Transformation of Cry IAc Gene Treatment Mediated by Agrobacterium tumefaciens

Research about transformation of cry IAc gene into subspesies Indica rice, Inpari 13, IR 64, and Inpari 6, had been conducted, but transgenic plant have not been yet obtained. Cry IAc is one of the group cry 1 gene with the highest toxin to pest of Lepidoptera especially stem borer. Transformation of cry IAc gene was conducted using Agrobacterium tumefaciens strain LBA4404 harboring plasmid pAY560325_cryIAc. Callus were grown in N6D induction media for 5 days before transformed by soaking in culture of Agrobacterium tumefaciens LBA4404. Subsequently, callus were grown in resting, selection, and regeneration medium. Cry IAc and hptII genes were confirmed through DNA extraction and PCR methods. Transformation of cry IAc gene successfully conducted and there were different responses of those varieties to tissue culture and transformation treatment. Inpari 13 showed the best response to transformation treatment based on the result of PCR of cry IAc and hptII genes, IR 64 gave the best response to tissue culture treatment based on the amount of embriogenic callus at callus induction medium."

"Latar Belakang: Insidensi infeksi jamur oportunistik yang disebabkan oleh Candida krusei terus meningkat. Di sisi lain, beberapa penelitian melaporkan adanya penurunan sensitivitas C. krusei terhadap caspofungin, vorikonazol, Amfoterisin B, flusitosin, dan ketokonazol. Selain itu, pilihan obat untuk infeksi Candida krusei menimbulkan berbagai efek samping. Oleh karena itu, diperlukan pengobatan alternatif yang lebih efektif dan aman, salah satunya adalah daun Polyscias scutellaria. Tujuan: Penelitian ini dilakukan untuk mengetahui efektivitas daun Polyscias scutellaria terhadap Candida krusei in vitro. etode: Penelitian eksperimental ini untuk menguji efektivitas daun Polyscias scutellaria terhadap Candida krusei in vitro dengan menggunakan metode difusi cakram dan dilusi. Konsentrasi daun Polyscias scutellaria yang digunakan adalah 800 mg/mL, 1600 mg/mL, 3200 mg/mL, 6400 mg/mL, dan 12800 mg/mL. Hasil: Ekstrak daun Polyscias scutellaria memiliki aktivitas fungistatik terhadap Candida krusei dengan nilai KHM 12800 μg/mL. Diskusi: Daun Polyscias scutellaria berpotensi sebagai antifungi terhadap Candida krusei. Pembacaan hasil setelah 24 dan 48 jam inkubasi dapat dilakukan pada penelitian selanjutnya untuk memberikan hasil yang lebih optimal.

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Background: The incidence of opportunistic fungal infection caused by Candida krusei has been increased. On the other hand, several researches had been reported the decrease sensitivity of Candida krusei to caspofungin, voriconazole, Amphotericin B, flucytosine, and ketoconazole. Moreover, drug of choice for Candida krusei infection cause various side effects. Therefore, it be required the alternative therapy that is more effective and safer, one of which is Polyscias scutellaria leaf. Objective: This research was done to determine the effectiveness of Polyscias scutellaria leaf to Candida krusei in vitro. Methods: This experimental study is to test the effectiveness of Polyscias scutellaria leaf against Candida krusei in vitro using disc diffusion method and dilution method. The extract concentrations of Polyscias scutellaria leaf that be used are 800 mg/mL, 1600 mg/mL, 3200 mg/mL, 6400 mg/mL, and 12800 mg/mL. Results: Polyscias scutellaria leaf extract has fungistatic activity to Candida krusei with MIC value is 12800 μg/mL. Discussion: Polyscias scutellaria leaf extract is potent as antifungal against C. krusei. The reading time after 24 and 48 hours incubation can be considered in the next research to provide more optimal results."

"Telah dilakukan penelitian yang bertujuan untuk melakukan transformasi gen cry IAc ke dalam kalus padi Inpari 13 dengan bantuan Agrobacterium tumefaciens serta mengetahui pengaruh kombinasi NAA+BAP dan NAA+Kinetin pada tahapan regenerasi. Gen cry IAc merupakan gen yang berperan dalam sintesis δ-endotoksin yang sifatnya sangat toksik pada serangga Lepidoptera. Gen cry IAc ditransformasi ke kalus padi Inpari 13 dengan menggunakan Agrobacterium tumefaciens strain LBA4404 yang membawa plasmid rekombinan pAY560325_cryIAc. Regenerasi kalus padi Inpari 13 hasil trasformasi dilakukan dengan penambahan kombinasi NAA+BAP dan NAA+Kinetin pada media regenerasi. Gen cry IAc berhasil ditransformasi dengan menggunakan metode Agrobacterium tumefaciens. Penggunaan kombinasi NAA+Kinetin menunjukkan regenerasi kalus yang lebih baik dibandingkan dengan kombinasi NAA+BAP.

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Research about transformation of cry IAc gene into Inpari 13 rice calli using Agrobacterium tumefaciens and Effect of Combination of NAA+BAP and NAA+Kinetin during regeneration process had been conducted. Cry IAc gene encodes δ-endotoxin which is highly toxic to Lepidoptera. Cry IAc gene was transformed into Inpari 13 rice calli using Agrobacterium tumefaciens strain LBA4404 containing recombinant plasmid pAY560325_cryIAc. Regeneration of transformed calli was grown in regeneration media that added by combination of NAA+BAP and NAA+Kinetin. Cry IAc gene had successfully been transformed using Agrobacterium tumefaciens method. The use of combination of NAA+Kinetin showed better calli regeneration than NAA+BAP."

"Telah dilakukan penelitian yang bertujuan untuk melakukan transformasi gen OsERA1 ke kalus padi cv. Taipei 309 menggunakan Agrobacterium tumefaciens. Gen OsERA1 adalah gen yang berperan dalam meningkatkan sensitifitas dari sel penjaga pada stomata terhadap asam absisat. Transformasi gen OsERA1 dilakukan menggunakan Agrobacterium tumefaciens strain LBA4404 yang membawa plasmid rekombinan pCAMBIA1301-OsERA1. Gen OsERA1 telah dikloning sebelumnya pada vektor pengklonaan pGEM-T Easy. Gen dipotong dengan enzim restriksi BamHI dan SalI. Gen diligasikan dengan pCAMBIA 1301 dan ditransformasikan ke dalam Escherichia coli DH5α dihasilkan vektor rekombinan pCAMBIA-ERA1. Plasmid vektor rekombinan pCAMBIA-ERA1 belum berhasil ditransformasi ke dalam Agrobacterium tumefaciens dengan elektroporasi tetapi berhasil dilakukan dengan particle bombardment.

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Research about transformation of OsERA1 gene into rice calli cv. Taipei 309 using Agrobacterium tumefaciens had been done. OsERA1 gene is a gene that has response to enhance sensitivity of guard cell to absicid acid. Transfer of OsERA1gene into calli was carried out using Agrobacterium tumefaciens strain LBA4404 harboring recombinant plasmid pCAMBIA1301-OsERA1. OsERA1 gene had been cloned previously in the pGEM-T Easy cloning vector and for inserting into pCAMBIA1301, the gene was digested from cloning vector using restriction enzymes BamHI and SalI. Furthermore, the gene was ligated into vector pCAMBIA 1301 and transformed into Escherichia coli DH5α in order to obtain recombinant plasmid pCAMBIA-OsERA1. The recombinant plasmid pCAMBIAOsERA1has not sucessfully transformed with electroporation into Agrobacterium tumefaciens yet, but it can be transformed by particle bombardment method."

"Telah dilakukan penelitian yang bertujuan untuk melakukan transformasi gen Osdep1-Tc ke kalus padi cv. Taipei 309 menggunakan Agrobacterium tumefaciens. Transformasi gen Osdep1-Tc dilakukan menggunakan A. tumefaciens strain LBA4404 yang membawa plasmid rekombinan pCAMBIA1301-Osdep1-Tc, mengandung gen reporter (gus), gen nptII dan hptI. Gen Osdep1-Tc yang telah dikloning ke vektor pengklonaan pGEM-T Easy pada penelitian sebelumnya digunakan sebagai sampel untuk kemudian isubkloning ke pCAMBIA 1301 dan ditransformasikan ke dalam Escherichia coli DH5α sehingga dihasilkan vektor rekombinan pCAMBIA-Osdep1-Tc. Vektor rekombinan kemudian dielektroporasi ke A. tumefaciens dan ditransformasi ke kalus embriogenik padi. Aktivitas GUS pada kalus berhasil dideteksi 3 hari setelah infeksi dengan A. tumefaciens. Analisis PCR kalus transforman menunjukkan bahwa gen hptI berhasil terintegrasi dengan stabil pada kelima kalus uji.

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Research about transformation of Osdep1-Tc gene into rice calli cv. Taipei 309 using Agrobacterium tumefaciens had been done. Transformation of Osdep1-Tc was carried out using A. tumefaciens strain LBA4404, harbored recombinant plasmid pCAMBIA1301-Osdep1-Tc, which contained a reporter gene (gus), hptI and nptII gene. Osdep1-Tc gene had been cloned previously into the pGEM-T Easy cloning vector. The gene was being subcloned into pCAMBIA 1301 and transformed into Escherichia coli DH5α in order to obtain recombinant vectors pCAMBIA-Osdep1-Tc. Furthermore, the recombinant vectors was electroporated into A. tumefaciens and transformed into rice embryogenic calli. GUS activity in rice calli was detected 3 days after infection with A. tumefaciens. PCR analysis of the transformant calli revealed that all five calli tested showed a succeeded stable integration of hptI gene."

"Dari semua spesies Candida yang ada, Candida krusei memiliki resistansi alami terhadap flukonazol, terapi lini pertama pada infeksi jamur. Salah satu faktor resistansi yang ada adalah adanya protein Abc1p yang dikode oleh gen ABC1. Penelitian eksperimental observasional ini menganalisis 16 whole-genome sequence (WGS) dari C. krusei dan membandingkannya terhadap gen ABC1 pada C. krusei lain dan homolognya pada Candid lain yang didapat dari database NCBI dan UniProtKB. Hasil analisis menunjukkan karakteristik dari gen Abc1p yang ditemukan memiliki common ancestor dengan gen serupa pada Candida lain. Hasil analisis juga memprediksi lokasi-lokasi pada sekuens yang diduga memiliki efek perubahan fungsi yang besar jika terjadi mutasi pada titik tersebut. Pemodelan 3D menemukan protein Pdr5 dari S. cerevisiae sebagai protein dengan struktur yang paling mirip dengan Abc1p.

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Of all the Candida species, Candida krusei has a natural resistance to fluconazole, the first line therapy for fungal infections. One of the factors that contributes to C. krusei’s resistance is the Abc1p protein that is coded by the ABC1 gene. This experimental observational study analyzed 16 whole-genome sequences (WGS) of C. krusei and compared them with the ABC1 genes of other C. krusei and its homologues in other Candida species gathered from the NCBI and UniProtKB databases. Results showed the characteristics of the Abc1p gene. A common ancestor among the ABC1p protein and other similar proteins in other Candida species was found. A prediction was also made on the effects an amino acid mutation would have and the location of the mutation. Three-dimensional modeling of the Abc1p protein showed that the protein with the most similar structure is the Pdr5 protein from S. cerevisiase."

"Sepsis merupakan suatu penyakit sistemik yang dapat disebabkan oleh translokasi bakteri. B. animalis subsp. lactis merupakan salah satu bakteri yang berpotensi dalam mencegah translokasi bakteri. Gen grpE merupakan salah satu target spesifik dalam mendeteksi B. animalis subsp. lactis. Penelitian bertujuan untuk mengoptimasi primer dengan target gen grpE untuk kurva standar, mengetahui hubungan konsentrasi primer terhadap nilai Ct, serta mengetahui sensitivitas dan spesifisitas primer. Isolat diisolasi dari sampel feses bayi menggunakan metode fenol-kloroform. Pasangan Primer dirancang berdasarkan sekuens gen grpE B. animalis subsp. lactis (NZ_ABOT01000010.1) menggunakan program Primer3. Optimasi primer dilakukan menggunakan lima konsentrasi berbeda,yaitu 50/50 nM, 100/100 nM, 300/300 nM, 500/500 nM, dan 1.000/1.000 nM.Hasil penelitian menunjukkan bahwa pasangan primer F_HNO19_grpE dan R_HNO19_grpE dengan konsentrasi 1.000/1.000 nM menghasilkan kurva standar yang optimal dengan efisiensi dan koefisien korelasi (R2) masing-masing sebesar 99,095% dan 0,971. Berdasarkan uji sensitivitas dan spesifistas, pasangan primer F_HNO19_grpE dan R_HNO19_grpE konsentrasi 1.000/1.000 nM dapat mengamplifikasi DNA target sampai dilusi 10-5 , tetapi spesifisitasnya hanya sampai dilusi 10-2. Konsentrasi primer tidak berkorelasi terhadap nilai Ct. Pasangan primer F_HNO19_grpE dan R_HNO19_grpE dapat digunakan untuk kuantifikasi B. animalis subsp. lactis dengan kisaran nilai Ct sebesar 15,74--33,89.

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Sepsis is a systemic disease that can be caused by bacterial translocation. B. animalis subsp. lactis is one of the bacteria that has the potential to prevent the bacterial translocation. grpE gene is a specific target in the detection of B.animalis subsp. lactis. The research aims to optimize primer pair with target gene grpE for generating standard curve, to know the correlation between primer concentration and Ct value, and to know the primer sensitivity and specificity. Isolates were isolated from infant stool samples using phenol-chloroform method. Primer pair is designed based on B. animalis subsp. lactis grpE gene sequence (NZ_ABOT01000010.1) using the Primer3 program. The primer optimization is done using five different concentrations, which are 50/50 nM, 100/100 nM,300/300 nM, 500/500 nM, and 1.000/1.000 nM. The results showed that the primer pair F_HNO19_grpE and R_HNO19_grpE with 1.000/1.000 nM concentration can be used to generate an optimal standard curve with efficiency and correlation coefficient (R2) each by 99.095% and 0.971. Based on sensitivity and specificity test, primer pair F_HNO19_grpE and R_HNO19_grpE with 1.000/1.000 nM concentration can amplify DNA targets up to 10-5 dilution, but its specificity is only up to 10-2 dilution. Primer concentration and DNA samples with different concentrations were not correlated to the Ct value. F_HNO19_grpE and R_HNO19_grpE primer pair can be used for quantification of B. animalis subsp. lactis with a range of Ct values of 15.74 to 33, 89."

"ABSTRAKMakanan modern di dunia saat ini banyak menggunakan pemanis dengan kadar fruktosa tinggi disertai makanan tinggi lemak yang meningkatkan resiko sindrom metabolik dan dapat menyebabkan perlemakan organ seperti ginjal. Ekstrak dari tanaman Acalypha indica Linn AI disinyalir memiliki efek protektif terhadap kerusakan jaringan pada perlemakan organ seperti hati. Penelitian ini bertujuan menguji efek protektif ekstrak AI pada perlemakan ginjal dengan hewan coba tikus. Tikus Sprague-Dawley dibagi secara acak ke dalam lima kelompok. Semua kelompok kecuali kelompok normal diinduksi dengan diet tinggi fruktosa dan lemak selama 4 minggu yang kemudian diberikan perlakuan berupa akuades, obat standar gemfibrosil G , ekstrak AI, dan kombinasi G AI pada masing-masing kelompok selama 4 minggu. Variabel yang diukur adalah keliling glomerulus, keliling tubulus, dan diameter tubulus pada ginjal. Perbedaan bermakna p < 0,05 hanya ditemukan pada variabel keliling tubulus antara kelompok tikus dengan terapi ekstrak AI dan kelompok kontrol negatif. Perbedaan yang tidak bermakna antara kelompok normal dan kontrol negatif menandakan gagalnya induksi perlemakan pada organ ginjal yang mungkin disebabkan duraksi induksi terlalu singkat. Penelitian ini menyimpulkan bahwa kombinasi ekstrak AI dan G tidak menunjukkan efek protektif terhadap perlemakan ginjal pada tikus.

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ABSTRAKModern foods these days often use high fructose sweetener and contain high level of fat which increase risk of metabolic syndrome and cause fat accumulation in organs such as kidney. An extract of Acalypha indica Linn AI was known to have protective effect against cell destruction on organs with fat accumulation such as liver. This research aimed to examine the protective effect of extract from Acalypha indica Linn on rats with fatty kidney. Male Sprague Dawley rats were randomized into five groups. All groups except normal groups were induced with high fructose and fat diets for 4 weeks and then were given aquades, standard medication gemfibrozil G , extract from AI, and combination of both G AI for another 4 weeks according to the group. Variables measured are glomerulus circumference, tubules circumference, and tubules diameter of kidney. Significant difference p 0.05 was found only in tubules circumference between group treated with AI and negative control group. Unsignificant difference between negative control and normal group indicate failure of fatty kidney induction probably caused by duration of induction that was too short. This research concludes that combination of AI and G does not show any protective effect on rat rsquo s fatty kidney."

"Instalasi Pengolahan Lumpur Tinja (IPLT) merupakan pengolahan air limbah yang dirancang hanya menerima dan mengolah lumpur tinja yang berasal dari sistem setempat yang diangkut melalui sarana pengangkutan lumpur tinja. Lumpur tinja yang dihasil tersebut tentu harus diolah terlebih dahulu agar sesuai dengan baku mutu yaitu, Peraturan Menteri Lingkungan Hidup Nomor 68 Tahun 2016 Tentang Baku Mutu Air Limbah Domestik. Namun demikian, dalam proses pengolahan air limbah ini, tidak dapat dihindari kemungkinan terlepasnya pencemar udara mikrobiologis (bioaerosol) ke udara sekitar. Tujuan penelitian ini untuk mengetahui sumber pencemar, mengetahui total bakteri dan jamur di udara serta perbedaan konsentrasi bakteri dan jamur pada musim kemarau dan musim hujan, dan meninjau faktor lingkungan (suhu, kelembaban, dan Kecepatan angin) yang mempengaruhi konsentrasi. Penelitian ini dilakukan pada musim kemarau dan hujan dengan masing-masing lima hari pengambilan dan dilakukan di empat titik pada IPLT Kalimulya Depok (unit bak pengisian, digester anaerob, pemekat lumpur, dan biofilter aerob-anaerob). Dari hasil penelitian, rata-rata konsentrasi bakteri pada musim kemarau yaitu unit bak pengisian sebesar 243±265 CFU/m3, pemekat lumpur sebesar 155±326 CFU/m3, digester anaerob sebesar 154±157 CFU/m3, dan biofilter aerob anaerob sebesar 76±122 CFU/m3. Sedangkan pada musim hujan konsentrasi bakteri yaitu unit bak pengisian sebesar 33±24 CFU/m3, pemekat lumpur sebesar 25±62 CFU/m3, biofilter aerob-anaerob sebesar 21±20 CFU/m3, dan digester anaerob sebesar 16±13 CFU/m3. Kemudian pada musim kemarau, konsentrasi jamur pada pemekat lumpur sebesar 516±554 CFU/m3, unit bak pengisian sebesar 364±202 CFU/m3, digester anaerob sebesar 340±181 CFU/m3, dan biofilter aerob-anaerob sebesar 231±201 CFU/m3. Sedangkan pada musim hujan konsentrasi jamur pada unit bak pengisian sebesar 58±39 CFU/m3, pemekat lumpur sebesar 55±33 CFU/m3, digester anaerob sebesar, 36±32 CFU/m3, dan biofilter aerob-anaerob sebesar 32±23 CFU/m3. Sehingga, diketahui konsentrasi bakteri tertinggi ditemukan pada unit bak pengisian pada musim kemarau dan terendah pada digester anaerob pada musim hujan. Konsentrasi jamur tertinggi ditemukan di pemekat lumpur pada musim kemarau dan terendah pada biofilter aerob-anaerob pada musim hujan. Konsentrasi bakteri dan jamur berada dibawah standar baku mutu. Sedangkan korelasi antara faktor lingkungan terhadap konsentrasi bakteri dan jamur ditemukan di beberapa tempat dan terdapat juga perbedaan konsentrasi bakteri dan jamur pada musim kemarau dan musim hujan.

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Sewage Treatment Plants (STPs) are wastewater processing systems that are designed to process only stool mud received from local systems of stool mud transport. The stool mud received must be processed so that it abides to the standard of quality according to the Regulation of the Minister of the Environment Number 68 Year 2016 concerning Domestic Wastewater Quality Standards. However, in the treatment process, there is a probability for a microbiological air pollutant (bioaerosol) to be produced that cannot be avoided. This research aims to analyze the source of pollution, the total amount of bacteria and fungi in the air, the difference of bacteria and fungi concentration between the dry and rainy season, and observe the environmental factors (temperature, humidity, wind speed) that affects bacteria and fungi concentration. This research was done during the dry and rainy season, each for a 5 day period in four observation points at the Kalimulya Depok STP (filling unit, anaerobic digester, mud concentrator and aerobic-anaerobic biofilter). The results of this research shows that the average bacteria concentration during the dry season is 243±265 CFU/m3 at the filling unit, 155±326 CFU/m3 at the mud concentrator, 154±157 CFU/m3 at the anaerobic digester, and 76±122 CFU/m3 at the aerobic-anaerobic biofilter. During the rainy season, the average bacteria concentration is 33±24 CFU/m3 at the filling unit, 25±62 CFU/m3 at the mud concentrator, 21±20 CFU/m3 at the aerobic-anaerobic biofilter, and 16±13 CFU/m3 at the anaerobic digester. The average fungi concentration during the dry season is 516±554 CFU/m3 at the mud concentrator, 364±202 CFU/m3 at the filling unit, 340±181 CFU/m3 at the anaerobic digester, and 231±201 CFU/m3 at the aerobic-anaerobic biofilter. As for the rainy season, the average fungi concentration is 58±39 CFU/m3 at the filling unit, 55±33 CFU/m3 at the mud concentrator, 36±32 CFU/m3 at the anaerobic digester, and 32±23 CFU/m3 at the aerobic-anaerobic biofilter. It can be seen that for the bacteria concentration, its highest value occurs at the filling unit during the dry season while its lowest value occurs at the anaerobic digester during the rainy season. For the fungi concentration, its highest value occurs at the mid concentrator during the dry season while its lowest value occurs at aerobic-anaerobic biofilter during the rainy season. The bacteria and fungi concentration values lie below the standard of quality. There are several correlations between environmental factors and the bacteria and fungi concentration values in some of the observed locations. There is also a difference between the bacteria and fungi concentration during the dry season and the rainy season.

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