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Afriyanti Sandhi
Abstrak :
[ABSTRAK
LATAR BELAKANG: Erythropoetin (EPO) sebagai hematopoietic growth factor, menarik perhatian para peneliti akibat efeknya dalam melindungi jaringan. EPO berinteraksi dengan vascular endothelial growth factor (VEGF) dan menstimulasi mitosis dan motilitas sel endotel dalam proses neo-angiogenesis; dan hal ini penting dalam fenomena kompleks penyembuhan luka, Tujuan penelitian ini adalah menyelidiki efek pemberian EPO pada penyembuhan luka bakar eksperimental di hewan coba. METODE: Lima belas ekor tikus Sprague-Dawley, strain dari Rattus Novergicus dengan berat antara 300-350 gram yang merupakan subjek hewan coba pada penelitian ini dibuat perlakuan eksperimental luka bakar grade 2B (dermis dalam). Lalu hewan coba akan dibagi ke dalam tiga grup secara acak dan mendapatkan terapi injeksi EPO dosis rendah (600 IU/mL), injeksi EPO dosis tinggi (3000 IU/mL) dan tidak mendapatkan perlakuan terapi apapun (grup kontrol). Setelah 14 hari observasi, dilakukan penilaian secara kuantitatif dari proses penyembuhan luka dengan menghitung persentasi epitelialisasi menggunakan perangkat lunak Analyzing Digital Images®. Dilakukan pula penilaian secara kualitatif dengan menghitung skor perubahan histopatologis pada penyembuhan luka. HASIL: Ukuran luka dan percepatan epitelialisasi dihitung pada hari ke-0, hari ke-5, hari ke-10 dan hari ke-14. Didapatkan bahwa hasil rerata ukuran raw surface (p value: 0.012 pada hari ke-5; 0.009 pada hari ke-10 and 0.000 pada hari ke-14) dan persentase penyembuhan luka (p value: 0.011 pada hari ke-5; 0.016 pada hari ke-10 and 0.010 pada hari ke-14), nilai terbaik dicapai oleh grup injeksi EPO dosis rendah. Evaluasi histopatologis menunjukkan bahwa skor tertinggi untuk re-epitelialisasi, jaringan granulasi dan neo-angiogenesis juga didapatkan pada grup injeksi EPO dosis rendah. SIMPULAN: Pada studi hewan coba menggunakan tikus Sprague-Dawley ini, didapatkan bahwa injeksi Recombinant Human EPO (rHuEPO) dapat mempercepat proses re-epitelialisasi dan penyembuhan luka yang disebabkan oleh luka bakar grade 2B (dermis dalam). Temuan ini diharapkan akan membuka pengetahuan baru dalam peningkatan kualitas terapi pada penyembuhan luka bakar.
ABSTRACT
BACKGROUNDS: The hematopoietic growth factor erythropoietin (EPO) attracts attention due to its all-tissue-protective pleiotropic properties. EPO interacts with vascular endothelial growth factor (VEGF) and stimulates endothelial cell mitosis and motility in neo-angiogenesis; thus it may of importance in the complex phenomenon of wound healing. The purpose of this study is to investigate the effect of EPO in experimental burn wounds healing. METHODS: Fifteen healthy Sprague-Dawley, strain of Rattus Novergicus weighing 300-350 grams, were prepared to achieve deep dermal burns. Animals were randomized to receive either low-dose EPO injection (600 IU/mL), high-dose EPO injection (3000 IU/mL) or nothing (control group). After 14 days of observations, a quantitative assessment of wound healing was determined by percentage of wound closure and epithelialization using Analyzing Digital Images® Software. And qualitative assessment was done to evaluate the score of histopathological changes in wound healing. RESULTS: The size of the wound area and re-epithelialization rate percentage was determined on Day-0, Day-5, Day-10 and Day-14. The average of raw surface areas measurement (p value: 0.012 in day-5; 0.009 in day-10 and 0.000 in day-14) and healing percentage of the lesions (p value: 0.011 in day-5; 0.016 in day-10 and 0.010 in day-14) were significantly best in the low- dose EPO group compared to the control group and high-dose EPO group. The histopathology evaluation revealed that the highest score for re-epithelialization, granulation tissue and neo- angiogenesis were achieved by the low-dose EPO injection group than in both control and high- dose EPO injection groups. CONCLUSIONS: In this animal study using Sprague-Dawley rats, Recombinant Human EPO (rHuEPO) injection administration prompted the evidences of improved re-epithelialization and wound healing process of the skin caused by deep dermal burns. These findings may lead to a new therapeutic approach to improve the clinical outcomes for the management of burns wound healing., BACKGROUNDS: The hematopoietic growth factor erythropoietin (EPO) attracts attention due to its all-tissue-protective pleiotropic properties. EPO interacts with vascular endothelial growth factor (VEGF) and stimulates endothelial cell mitosis and motility in neo-angiogenesis; thus it may of importance in the complex phenomenon of wound healing. The purpose of this study is to investigate the effect of EPO in experimental burn wounds healing. METHODS: Fifteen healthy Sprague-Dawley, strain of Rattus Novergicus weighing 300-350 grams, were prepared to achieve deep dermal burns. Animals were randomized to receive either low-dose EPO injection (600 IU/mL), high-dose EPO injection (3000 IU/mL) or nothing (control group). After 14 days of observations, a quantitative assessment of wound healing was determined by percentage of wound closure and epithelialization using Analyzing Digital Images® Software. And qualitative assessment was done to evaluate the score of histopathological changes in wound healing. RESULTS: The size of the wound area and re-epithelialization rate percentage was determined on Day-0, Day-5, Day-10 and Day-14. The average of raw surface areas measurement (p value: 0.012 in day-5; 0.009 in day-10 and 0.000 in day-14) and healing percentage of the lesions (p value: 0.011 in day-5; 0.016 in day-10 and 0.010 in day-14) were significantly best in the low- dose EPO group compared to the control group and high-dose EPO group. The histopathology evaluation revealed that the highest score for re-epithelialization, granulation tissue and neo- angiogenesis were achieved by the low-dose EPO injection group than in both control and high- dose EPO injection groups. CONCLUSIONS: In this animal study using Sprague-Dawley rats, Recombinant Human EPO (rHuEPO) injection administration prompted the evidences of improved re-epithelialization and wound healing process of the skin caused by deep dermal burns. These findings may lead to a new therapeutic approach to improve the clinical outcomes for the management of burns wound healing.]
2015
T-Pdf
UI - Tesis Membership  Universitas Indonesia Library
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Resky Dwi Cahyati
Abstrak :

Xilanase merupakan enzim yang dapat mendegradasi xilan menjadi xilooligosakarida berbagai ukuran dengan memotong ikatan 1,4-β-D-xylosidik dan memiliki potensi tinggi dalam aplikasi industri. Pada penelitian sebelumnya, untuk produksi xilanase pendekatan teknologi DNA rekombinan telah dilakukan, yaitu dengan mengkonstruksi Pichia pastoris yang telah dimodifikasi genetik sehingga dapat memproduksi xilanase dari Bacillus halodurans CM1. Peningkatan produksi xilanase menggunakan starter Pichia pastoris telah dilakukan. Namun, karena tingginya biaya medium standar produksi dengan Pichia pastoris pada skala laboratorium dan bioreaktor, maka komposisi media standar disubtitusi dengan medium minimal. Akan tetapi aktivitas xilanase yang dihasilkan relatif rendah. Oleh karena itu dalam penelitian ini dilakukan modifikasi medium berbiaya lebih rendah, dimana gliserol murni disubtitusi dengan gliserol teknis sebagai sumber C, sedangkan sumber N organik yaitu pepton dan yeast ekstrak masing-masing disubtitusi dengan hidrolisat tepung kedelai (18% (b/v) total kandungan N) dan dedak padi (bekatul) (14,63% (b/v) total kandungan N), dan amonium sulfat sebagai tambahan sumber N anorganik yang dioptimasi untuk menghasilkan xilanase dengan harapan biaya yang digunakan lebih murah. Penggunaan gliserol teknis 1% (v/v) dan campuran hidrolisat tepung kedelai 15 g/100 mL, hidrolisat dedak padi 30 g/100 mL,dan amonium sulfat 2,5% (b/v) ditemukan sebagai media sesuai yang menghasilkan tingkat tertinggi aktivitas xilanase (1383,898 U/mL), aktivitas spesifik (861,705 U/mg), kadar protein (1,606 mg/mL), dan berat sel kering (43,300 g/L). Penambahan konsentrasi amonium sulfat meningkatkan produksi xilanase rekombinan sekitar hampir dua kali lipat, dengan persen peningkatan sebesar 97,46%


Xylanase is an enzyme that can degrade xylan into xylooligosaccharides of various sizes using 1,4-β-D-xylosidik bonds and has high potential in industrial applications. In the previous study, to produce xylanase using recombinant DNA technology was done, namely by constructing Pichia pastoris which has facilitated genetics so that it can produce xylanase from Bacillus halodurans CM1. The increase in xylanase production using starter has been done by Pichia pastoris. However, due to the high cost of standard production with Pichia pastoris on a laboratory scale and bioreactor, the composition of standard media is substituted with a minimum medium. However, the xylanase activity produced is relatively low. Therefore in this study modification of the lower cost medium, pure glycerol was substituted with technical glycerol as karbon source, while the organic nitrogen source is peptone and yeast extract substituted with soybean hydrolyzate (18% (b/v) total N content) and rice bran (14.63% (b/v) total N content), respectively,and ammonium sulfate as an additional source of nitrogen inorganic which was optimized to produce xylanase in the hope that costs are cheaper. The use of technical glycerol 1% (v/v) and a mixture of 15 g/100 mL soybean hydrolyzate, rice bran hydrolyzate 30 g/100 mL, and 2.5% (b/v) ammonium sulfate were found to be suitable media that yielded profit high xylanase activity (1383,898 U/mL), specific activity (861.705 U/mg), protein content (1,606 mg/mL, based bradford method), and dry cell weight (43,300g/L). The addition of ammonium sulfate concentrations increased the production of recombinant xylanase by almost double, with a percent increase of 97,46%

Depok: Fakultas Matematika dan Ilmu Pengetahuan Alam, 2020
T55461
UI - Tesis Membership  Universitas Indonesia Library
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Indri Aderni
Abstrak :
Latar belakang: Beta defensin diekspresikan terutama oleh sel epitel pada permukaan mukosa berbagai organ seperti kulit, usus, mulut dan saluran genital. Studi sebelumnya menunjukkan bahwa Beta defensin 30 (Defb30) terekspresi spesifik di epididimis. Defb30 merupakan peptida kationik berukuran kecil yang diduga berperan penting pada proses pematangan spermatozoa di epididimis dan juga memiliki kemampuan untuk membunuh mikroba. Untuk mempelajari aktivitas antimikroba Defb30 ini diperlukan analisis pada tingkat protein dan hal tersebut memerlukan protein dalam jumlah yang cukup. Karena itu perlu dilakukan suatu rekayasa genetika berupa perancangan gen yang mengkode Defb30, pengklonaan dan ekspresi untuk pembuatan protein rekombinan DEFB30. Metode: Gen sintetik penyandi protein DEFB30 yang telah dioptimasi kodonnya diklona ke dalam vektor pQE-80L. Plasmid rekombinan yang mengandung sisipan gen target dikonfirmasi dengan analisis enzim restriksi dan sekuensing untuk selanjutnya diekpresikan ke dalam E. coli BL21 dan diinduksi menggunakan IPTG (Isopropyl-1-Thio-d-Galactopyranoside) dengan berbagai waktu inkubasi. Deteksi protein rekombinan dilakukan dengan SDS-PAGE dan westernblotting. IMAC (Immobilized Metal Affinity Chromatography) digunakan untuk mempurifikasi protein rekombinan. Uji antimikroba protein rekombinan dilakukan dengan cara pengukuran nilai optical density (OD) dan dianalisis hasilnya menggunakan uji one way anova. Hasil: Gen sintetik penyandi protein rekombinan DEFB30 berhasil dikonstruksi pada plasmid pQE-80L. Ekspresi ke dalam E. coli BL21 menghasilkan suatu protein fusi setelah diinduksi menggunakan IPTG selama 4 jam. Hasil analisis protein rekombinan dengan westernblotting menggunakan antibodi Anti-His G-HRP menunjukkan terbentuk pita tebal yang berukuran diatas 10 kDa (±12 kDa). Uji antimikroba protein rekombinan DEFB30 menunjukkan bahwa protein tersebut dapat menghambat pertumbuhan bakteri Eschericia coli dan Bacillus subtilis. Kesimpulan: Gen sintetik penyandi beta defensin 30 berhasil diklona ke dalam plasmid pQE-80L. Ekspresi protein rekombinan DEFB30 menghasilkan suatu protein fusi berukuran ±12kDa. Protein rekombinan DEFB30 terbukti memiliki sifat antimikroba terhadap Eschericia coli dan Bacillus subtilis. ......Background: Beta defensins are primarily expressed by epithelial cells at mucosal surfaces, such as those in skin, gut, mouth and genital tract. Previous studies have demonstrated that beta defensin 30 (Defb30) is exclusively expressed in the epididymis. Defb30 is known as a small cationic antimicrobial peptide which plays an important role in epididymal sperm maturation and also acts as a host defence against microbial infection. Study of Defb30 role in the antimicrobial activity requires generating DEFB30 protein for characterization. For the purpose of this study, Defb30 gene was designed, synthesized, cloned, and expressed for the manufacture of the DEFB30 recombinant protein. Method(s): In this study, according to the preferred codon in E. coli, the Defb30 gene was optimized and synthesized. The gene was cloned into pQE-80L vector and subsequently expressed in E. coli BL21; using IPTG (Isopropyl-1-Thio-d-Galactopyranoside) as an inducer. Detection of recombinant protein was carried out by using SDS-PAGE and westernblotting. IMAC (Immobilized Metal Affinity Chromatography) was used to purify recombinant protein. Optical density measurement was used to analyze antimicrobial property of the DEFB30 recombinant protein. Results: The synthetic gene was successfully constructed into pQE-80L plasmid and expression of the recombinant protein in E. coli BL21 produced a fusion protein after being induced by IPTG for 4 hours. Westernblotting analysis using Anti-His G-HRP antibody showed band above 10kDa (±12kDa). Antimicrobial assay for DEFB30 recombinant protein showed inhibition towards growth rates of Eschericia coli and Bacillus subtilis. Conclusion: Defb30 synthetic gene was succesfully cloned into pQE-80L plasmid. Expression of recombinant DEFB30 produced a fusion protein of ±12kDa. This recombinant protein has antimicrobial property towards Eschericia coli and Bacillus subtilis.
Depok: Fakultas Kedokteran Universitas Indonesia, 2020
T-Pdf
UI - Tesis Membership  Universitas Indonesia Library
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Syarif Safarudin
Abstrak :
Hemodialisis merupakan terapi pengganti fungsi ginjal terbanyak pada pasien gagal ginjal tahap akhir. Pasien yang menjalani terapi hemodialisis rutin sering mengalami penurunan kualitas hidup. Penelitian ini bertujuan menganalisis hubungan pola terapi, nilai ureum-kreatinin plasma dan hemoglobin dengan kualitas hidup pasien hemodialisis. Desain menggunakan cross sectionaldengan consecutive sampling terhadap 62 responden yang menjalani hemodialisis rutin di RSUD Dr. Soedarso Pontianak. Penilaian kualitas hidup dengan menggunakan kuesioner SF-36. Data menggunakan hasil regresi linier bergandamenunjukkan ada hubungan signifikan (p<0,05) antara durasi, frekuensi, terapi eritropoetin,nilai ureum-kreatinin plasma, hemoglobin, dan keputusasaan dengan kualitas hidup. Perawat perlu meningkatkan kualitas asuhan keperawatan untuk meningkatkan kualitas hidup pasien hemodialisis. ......Hemodialysis is the most renal replacement therapy for end stage renal disease. Patients undergoing regular hemodialysis often experience decreased in quality of life. The aim of this study was to analyze the relationship between pattern of therapy, urea-creatinine level of plasma and hemoglobin with quality of life patients undergoing hemodialysis. Research design used is cross sectional with consecutive sampling to 62 respondents underwent regular hemodialysis at Dr Soedarso general hospital. Quality of life was measured using SF-36 questionnaires. Data using the multiple linear regression showed no significant relationship (p <0.05) between duration, frequent, Erythropoietin therapy, urea-creatinine level of plasma, hemoglobin and hopelessness with patients quality of life. Nurses need to enhance quality of nursing care to improve the quality of life for the patients undergo hemodialysis.
Depok: Fakultas Ilmu Keperawatan Universitas Indonesia, 2012
T30441
UI - Tesis Open  Universitas Indonesia Library