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Abstrak :
Ruang Lingkup dan Cara : Gel cincau hijau memiliki sifat seperti agar sehingga menimbulkan pemikiran apakah dapat dipakai sebagai medium elektroforesis. Kenyataan ba,h a gel dapat terbentuk tanpa melalui proses pemanasan dianggap sebagai suatu keunggulan. Namun terdapat masalah apakah warna hijau disebabkan klorofil dapat dibuang, dan karakteristik gel belum diketahui dengan baik. Gel cincau dibuat dari daun Stephania hernandifolia, dan beberapa karakteristiknya dipelajari seperti : kondisi pembuatan, daya tahan /perubahan dalam penyimpanan pada berbagai suhu, dan upaya untuk menghilangkan klorofil. Kekuatan gel dinilai dari kemampuannya menahan beban (buatan sendiri) dan dengan curdmeter. Analisis kualitatif terhadap bubuk gel dilakukan untuk menentukan karbohidrat pembentuk gel serta monomernya (uji Molisch, jodium, Benedict, Barfoed, Bial, Tauber, osazon). Dilakukan pula upaya untuk menilai adanya enzim pektin esterase (produk : asam asetat), yang dilaporkan memegang pcranan dalam pembentukan gel. Hasil dan kesimpulan : Gel dengan konsistensi yang baik diperoleh dari 5 g daun segar dan 50 ml air. Pada suhu kamar (30°C) gel dapat bertahan sampai 3 hari; selanjutnya terjadi pengeluaran cairan (sineresis) yang berlangsung lebih cepat pada suhu lebih tinggi. Gel mampu menahan beban 20 g, dan dengan alas curdmeter 95,93 g (nilai untuk agar swallow 1% : 45 g dan 154,72 g). Etanol absolut dapat dipakai untuk mengekstraksi kiorofil dari gel, yang setelah dikeringkan, meninggaikan bubuk berwarna keabu-abuan yang tidak lagi mampu membentuk gel (ireversibel). Gel yang dikeringkan sampai berat konstan (3 hari , 70°C) menunjukkan kandungan bahan padat 0,147% (0,35731242,0815 g). Analisis kualitatif menyatakan karbohidrat pembentukan gel mengandung heksosa, pentosa dan asam uronat. Aktivitas enzim pektin esterase tidak berhasil diidentifikasi. Gel cincau belum berhasil digunakan sebagai medium elektroforesis antara lain disebabkan warna hijau belum dapat dihilangkan tanpa merusak sifat dan kemampuan membentuk gel, dan gel mengecil akibat panas yang terbentuk pada saat elektroforesis.

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Cincau Gel (Stephania hernandifalia) : Some Characteristics and The Possibility for use as Medium for ElectrophoresisScope and methods: It was thought that green cincau gel, on account of its agar-like property, could be used as a medium for electrophoresis. The fact that cincau gel could form without involving heat treatment., in contrast with agar, is considered an advantage. However, the properties of green cincau have not been well characterized, and the green color due to the presence of chlorophyll has to be removed. The gel, prepared from the leaves of Stephania hernandifolia, was studied for : preparative c9ndition, stability/changes during storage at different temperatures, and method for removing chlorophyll. Gel strength was evaluated from its ability to hold weight and by the use of curdmctcr. The gel powder, after removal of chlorophyll and drying to constant weight, was analysed for its carbohydrate constituents (test : Molisch, iodine, Benedict, Barfoed, Bial, Tauber, ozason). The presence of pectin esterase, which has been reported to be involved in the gelling process, was also examined (product : acetic acid). Findings and Conclusion: Green gel of good consistency was made from 5 g of leaves and 50 ml water. The gel was stable under storage for 3 days at room temperature (34°C). Water loss (syneresis) was observed during longer storage, and was more pronounced with increasing temperatures. The gel could withstand 20 g weight, or 95,93 by using a curdmotor (respective findings for 1% agar, 45 g and 154,72 g) Absolute ethanol was use to remove chlorophyll from the gel, however, the powder let after drying the gel was grayish in color and could no longer be reconstituted (irreversible). The decolorized gel, dried to constant weight (3 days, 70°C) conteined 0.147% solid (0.3573/242.0815). Qualitative analyses indicate the presence of hexose, pentose and uronic acid. Pectin esterase activity could not be detected. The gel could not yet be used as support material for electrophoresis due others, failure to remove chlorophyll without loss of the gel-forming ability, and shrinkage caused by the heat produced during an electhroporetic run.

Abstrak :
Latar belakang: Sitoglobin (Cygb) adalah protein pengangkut O2 yang diekspresikan oleh fibroblas dan fibroblast like cells aktif. Keperluan O2 dan energi meningkat pada fibrosis akibat proliferasi fibroblas dan sintesis kolagen. Pada fibrosis terjadi hipoksia yang ditandai oleh stabilisasi hypoxia inducible factor-1α (HIF-1α), yang kemudian membentuk HIF-1 yang merupakan faktor transkripsi untuk ekspresi protein adaptasi (termasuk Cygb). Diduga Cygb berperan dalam suplai O2 pada fibrosis. Tujuan penelitian ini adalah untuk memperoleh informasi mengenai peran Cygb pada hipoksia jaringan fibrosis dengan keloid sebagai model. Metode: Penelitian bersifat observasional deskriptif. Sampel keloid diperoleh melalui biopsi, sedangkan kontrol preputium diperoleh melalui sirkumsisi, masing-masing 10 sampel jaringan. Pengukuran ekspresi mRNA Cygb, HIF-1α, kolagen I dan III dilakukan dengan real time RT-PCR; kadar protein Cygb dan HIF-1α dengan ELISA; dan ekspresi protein Cygb, HIF-1α, FGF, kolagen I dan III di lapisan dermis dengan imunohistokimia (IHK). Pengukuran kadar MDA dan GSH (tingkat stres oksidatif) serta kadar hidroksiprolin (untuk pematangan kolagen) dengan spektrofotometri, sedangkan pengukuran kepadatan kolagen dengan pewarnaan Van Gieson. Data dianalisis secara statistik menggunakan uji-t. Hasil: Pada keloid dibandingkan preputium, ekspresi mRNA Cygb meningkat 8,7 kali, protein Cygb meningkat bermakna (1,196 Vs 0,779 ng/mg protein dan 95% Vs 63% ; p <0,05). Ekspresi mRNA HIF-1α meningkat 5,1 kali, protein HIF-1α meningkat bermakna (0,201 Vs 0,122 ng/mg protein dan 80% Vs 38%; p <0,05). Terdapat korelasi kuat antara ekspresi protein HIF-1α dan mRNA Cygb (Pearson; R = 0,649; p <0,01). Ekspresi protein FGF keloid meningkat bermakna (78% Vs 41%; p <0,05). Demikian pula ekspresi mRNA prokolagen I dan III keloid meningkat bermakna (35 kali dan 27,1 kali), serta ekspresi protein kolagen I dan III (61% Vs 37% dan 39% Vs. 16%; p <0,05). Juga terdapat korelasi kuat antara protein HIF-1α dengan FGF, prokolagen I dan III (Pearson; R= 0,878; R=0,960; dan R=0884; p<0,01). Kadar hiroksiprolin lebih tinggi pada keloid (0,297 Vs 276 ng/mg protein; p >0,05) dan pematangan kolagen lebih tinggi bermakna (1,2 kali; p <0,05). Cygb berkorelasi kuat dengan pematangan kolagen (kadar hidroksiprolin) (Pearson; R = 0,790; p <0,001). Kesimpulan: Cygb berperan pada hipoksia jaringan fibrosis yang ditandai dengan peningkatan ekspresinya. Peran Cygb terkait dengan ekspresi HIF-1α yang berkorelasi dengan peningkatan FGF, pro/kolagen I dan III yang merupakan faktor penting pada fibrosis. Cygb juga berperan pada pematangan kolagen.

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Background: Cytoglobin (Cygb) is an O2 carrier protein expressed by fibroblasts and active fibroblast like cells. O2 and energy demand increased in fibrosis due to proliferation of fibroblasts and synthesis of collagen. In fibrosis hypoxia occurred which is characterized by stabilization of hypoxia inducible factor-1α (HIF-1α), which later forming the HIF-1, a transcription factor for the expression of adaptation protein (including Cygb). Cygb alleged role in the supply of O2 in fibrosis. The purpose of this study was to obtain information about Cygb role in fibrosis hypoxia with keloid tissue as a model. Methods: This was an observational descriptive study. Keloid samples were obtained from biopsy, while the preputium as control were obtained from circumcision, 10 tissue samples each. Measurement of Cygb, HIF-1α, collagen I and III mRNA expression were carried out by real time RT?PCR. Cygb and HIF-1α protein level were measured by ELISA; while Cygb, HIF-1α, FGF, and collagen I and III protein expressions in the dermis layer by immunohistochemistry (IHC). Measurement of MDA and GSH levels (oxidative stress) and hydroxyprolin concentration (marker of mature collagen) by spectrophotometry, while the collagen density measurement with van Gieson staining. Data were analyzed statistically using t-test. Results: In keloid compared preputium, Cygb mRNA expression increased 8.7 times compared to preputium, Cygb protein increased significantly (1.196 Vs 0.779 ng/mg protein and 95% Vs 63%, p <0.05). HIF-1α mRNA expression increased by 5.1 times in keloid tissue, and protein HIF-1α increased significantly (0.201 Vs 0.122 ng/mg protein and 80% Vs 38%, p <0.05). There is a strong correlation between the expression of HIF-1α protein and Cygb mRNA (Pearson; R = 0.649, p <0.01). Keloid FGF protein expression increased significantly (78% Vs 41%; p <0.05). Similarly, mRNA expression of procollagen I and III keloid increased significantly (35 times and 27.1 times), and protein expression of collagen I and III (61% Vs 37% and 39% Vs 16%, p <0.05). There is also a strong correlation between HIF-1α protein with FGF, procollagen I and III (Pearson, R = 0.878, R = 0.960; and R = 0.884, p <0.01). Hydroxyprolin concentration were higher in keloid (0.297 Vs 0.276 ng/mg protein; p >0.05) and collagen maturation was significantly higher (1.2 times, p <0.05). Cygb is correlated with maturation of collagen (hydroxyproline levels) (Pearson, R = 0.790, p <0.001). Conclusion: Cygb play role in fibrosis hypoxia which is characterized by its increased expression. Cygb role is associated with the expression of HIF-1α which are correlated with increased FGF, pro/collagen I and III, which are important factor in fibrosis. Cygb also play a role in the maturation of collagen.