Identifikasi Sel-sel Target Virus Penyakit Jembrana dengan Teknik Imunositokimia Ganda

Penelitian bertujuan untuk mengidentifikasi sel target virus Jembrana dengan teknik imunositokimia ganda. Penelitian ini menggunakan sapi Bali yang diinokulasi dengan virus Jembrana secara intramuskuler. Pada demam hari kedua setelah inokulasi virus, sapi dinekropsi. Limpa diambil secara aseptik, kemudian direndam dalam buffer formalin 10% selama 24 jam. Potongan limpa diproses untuk pembuatan sediaan histologis dengan menggunakan cryomicrotome. Preparat histologis limpa diwarnai dengan teknik imunositokimia ganda. Untuk mengidentifikasi subset limfosit digunakan antibody monoclonal anti BoCD4 + dan anti BoCD8 + , serta diamino benzidine (DAB) sebagai substrat. Pada pewarnaan ini, sel-sel terinfeksi akan tampak berwarna biru, sedangkan sel-sel marka BoCD4 + atau BoCD8 + akan tampak berwarna coklat. Untuk identifikasi sel-sel terinfeksi virus Jembrana digunakan antibodi monoklonal anti-capsid JDV (BB-Vet Denpasar) dan nitro blue tetrazolium (NBT) sebagai substrat. Hasil penelitian menunjukkan bahwa sel-sel yang terinfeksi virus Jembrana hanya bermarka BoCD4 + , dan sama sekali tidak pada sel BoCD8 + . Simpulan penelitian ini adalah sel-sel BoCD4 + merupakan sel target virus Jembrana

Role of Isoflavones on Lipid Peroxidation, Superoxide Dismutase in Lymphocytes under Oxidative Stress Conditions

Oxidative stress conditions forms free radicals in cells. Free radicals react continuously and cause cell membranes damage including lymphocytes, thus decreasing cell function. Therefore, there were efforts need to be done to overcome abnormalities and decreased lymphocyte cell function under conditions of oxidative stress. One of them using bioactive compounds that have antioxidant activity, such as isoflavones. Isoflavones have been reported to have antioxidant activity and the potential to protect cells of oxidative stress. This study aims to determine the role of isoflavones as antioxidative in protecting lymphocytes under conditions of oxidative stress, be reviewed of CD4, CD8, malondialdehyde levels and superoxide dismutase activity. This study was used rat lymphocyte cell cultures in two conditions, i.e: oxidative stress and without oxidative stress. The cell culture were treated with 20 ml of 2 mM paraquat and  isoflavone at dose 0, 1000, 2000, and 3000 ppm, respectively. The parameters were observed: (1) malondialdehyde  (MDA) levels,  (2) superoxide dismutase (SOD) activity, and (3) percentage of CD4 and CD8 lymphocytes cell using a monoclonal anti-rat CD4 and CD8 FITC-conjugate. The results showed paraquat administration in cell culture on oxidative stress conditions could reduce the number of CD4 and CD8 lymphocytes cels, increased of malondialdehyde (MDA) and superoxide dismutase (SOD) activity. Administration of isoflavones on culture that treated without oxidative stress condition could increase percentage number of CD4, CD8, MDA levels, and SOD activity. Whereas, administration of isoflavones on oxidative stress condition prevented the decrease percentage number of CD4, CD8, MDA levels, and SOD activity. Administration of isoflavone at dose 3000 ppm in cell culture that treated on oxidative stress condition provided optimal results in preventing the decrease number of CD4, CD8 lymphocytes cell, MDA levels, and SOD activity. Keywords: isoflavone, lymphocytes, CD4, CD8, malondialdehyde, superoxide dismutase

Antioxidant Capacity Of Frangipani (Plumeria Alba) Powder Extract

This research aimed to identify the antioxidant capacity, vitamin C (ascorbic acid), and total phenolic compounds of frangipani flower powder. The powder was extracted using ethanol, methanol, acetic acid, and water (aquadest). Antioxidant capacity of each extract were determined using 2,2-diphenyl-1-picrylhydrazyl radical scavenging method, vitamin C were determined using 2,4 Dinitrophenylhydrazine, and total phenolic compounds were determined using Folin Ciocalteu reagent. All of parameters were measured by spectrofotometer. The result shows that the highest value of antioxidant capacity was ethanolic extract (18.19%) and the lowest value was acetic acid extract (12.74%). The highest value of vitamin C was aqueous extract (3.49 mg/100g) and the lowest value was acetic acid extract (3.02 mg/100g). The highest value of total phenolic content was aqueous extract (25.49 mg GAE/g) and the lowest value was acetic acid extract (22.74 mg GAE/g). In conclusion, the higher antioxidant capacity was not always followed by the higher of vitamin C and total phenolic compounds

Microbiological and Chemical Properties of Kefir Made of Bali Cattle Milk

Information regarding to microbiological and chemical characteristics, and incubation time is crucial in developing kefir prepared using Bali cattle milk. This study was intended to investigate microbiological and chemical properties of the kefir prepared of Bali Cattle milk and Indonesian kefir grains after 24, 48 and 72 hours incubation periods. A completely randomized design, with 3 treatments, and 9 replicates were undertaken. Kefir samples were taken at the end of incubation period for determination of total lactic acid bacterial and yeast counts, pH, titratable acidity, lactose percentage and protein content. The result of this research showed that the total lactic acid bacterial counts were 108 – 109 cfu/ml, while yeast counts were ranging from 105 – 106 cfu/ml, no coliform and Escherichia coli were detected in any kefir samples in this research. Identification of the lactic acid bacteria and yeast revealed that the Lactobacillus paracasei ssp. paracasei 1 was the predominant species found in the kefir samples, followed by Lactobacillus brevis and the yeast Candida famata. Chemical analysis of the kefir samples showed that, protein, lactose, titratable acidity and pH of the kefir samples were in the range of 5.68 - 6.26%, 3.98 - 4.67%, 0.89 - 1.73% and 3.38 - 4.35, respectively. The result also indicated that the incubation period significantly affected the microbial counts and chemical composition of Bali Cattle milk kefir. Keywords: Kefir, Bali cattle milk, Lactic acid bacteria, Yeast, Protein, Lactos

Hubungan Positif antara Ulkus Kaki Diabetik dengan Persentase Sel Bermarkah Cd4+ Pembawa Malondialdehid

Tingginya angka kejadian ulkus kaki diabetik (UKD) dan luka di kaki yangsulit sembuh memberi petunjuk kemungkinan ada proses kematian sel imun yangsangat banyak dan belum jelas mekanismenya secara molekuler pada jaringan UKD.Telah diteliti hubungan antara derajat UKD dengan persentase sel bermarkahCD4+ pembawa malondialdehid (MDA). Penelitian ini adalah penelitianobservasional dengan rancangan cross sectional analytic study yang dilakukan diRumah Sakit pemerintah dan swasta di Denpasar, Badung, Tabanan, dan Gianyar.Parameter yang diukur dari bahan darah adalah kadar gula darah memakai metodeenzimatik (heksokinase), dan dari bahan jaringan kaki, dihitung sel bermarkah CD4+pembawa MDA memakai metode imunohistokimia (reagen dari Biodesign danAbcam ). Dari 80 sampel UKD didapatkan 49 (61,2%) penderita laki-laki dan 31(38.8%) penderita wanita, berdasarkan tingkat keparahan UKD, sampel dipilah lagimenjadi: 29 (31,9%) derajat 2; 20 (21,9%) derajat 3; 13 (14,3%) derjat 4; dan18(19,8%) derajat 5, rata-rata persentase sel bermarkah CD4+MDA adalah 75,0 ±20,5 %, Didapatkan korelasi positif kuat antara persentase sel bermarkah CD4+pembawa malondialdehid dengan derajat UKD (r = 0,71; p < 0,01). Pada penelitianini membuktikan ada mekanisme kematian sel imun dan sekaligus menjawabpermasalahan bahwa pada penderita UKD mudah terkena infeksi dan sulit untukdisembuhkan, dengan dibuktikan bahwa ada korelasi positif kuat antara derajat UKDdengan persentase pembentukan MDA dari sel bermarkah CD4+, ini menyatakanbahwa semakin berat derajat UKD semakin banyak mengalami kematian sel imun.

Respon Kekebalan Humoral Mencit Balb/c yang Divaksinasi dengan Vaksin Limpa dan Vaksin Kultur Penyakit Jembrana terhadap Protein Virus Jembrana

Sapi Bali adalah salah satu aset nasional Indonesia yang harus dilestarikan karena mempunyai keuntungan ekonomi. Tetapi sapi Bali mempunyai beberapa kelemahan penyakit khususnya penyakit Jembrana yang disebabkan oleh virus penyakit Jembrana (JDV). Pencegahan terhadap penyakit Jembrana telah dilakukan dengan vaksinasi. Vaksin yang terbukti dapat menurunkan tingkat kematian sapi Bali terserang JDV adalah vaksin limpa. Jenis vaksin ini hanya mampu menginduksi kekebalan dengan perlindungan 70%. Proteksi ini dapat ditingkatkan jika jumlah virus yang digunakan dalam vaksin meningkat. Tekhnik kultur in vitro adalah salah satu metode meningkatkan jumlah virus penyakit Jembrana, dan selanjutnya dibuat vaksin kultur. Hasil penelitian menunjukkan bahwa sel limfosit sapi Bali terinfeksi JDV adalah 9,5% pada limpa dan 57,43% pada sel kultur. Uji westernimmunoblotting sel limfosit sapi Bali dari darah tepi dan limpa terinfeksi JDV menggunakan antibodi monoklonal (AbMo) anti Ca, terdeteksi protein dengan berat molekul 26 kDa, 42 kDa dan 51 kDa. Pada medium kultur PBMC dan endapan plasma sapi Bali terinfeksi JDV, teridentifikasi protein dengan berat molekul 16 kDa an 26 kDa menggunakan AbMo, dan teridentifikasi protein dengan berat molekul 16 kDa; 21,5 kDa. 26 kDa; 29,7 kDa; 40 kDa dan 50 kDa menggunakan AbPo. Uji Elisa didapatkan nilai absorban antibodi mencit balb/c yang divaksinasi dengan vaksin kultur penyakit Jembrana lebih tinggi yaitu sebesar 0,3089 dibandingkan vaksin limpa yaitu sebesar 0,177 dengan p<0,05. Nilai absorban antibodi mencit balb/c terhadap antigen Ca, SU dan tat, memperlihatkan nilai absorban terhadap antigen SU berbeda sangat signifikan dibandingkan dengan antigen Ca dan antigen tat (p<0,01). Antigen Ca berbeda signifikan terhadap antigen tat (p<0,05)

The role of caspase-dependent and caspase-independent pathways of apoptosis in the premature rupture of the membranes: A case-control study

Background: Premature rupture of membrane (PROM) remains a problem in obstetrics, the mechanisms of PROM have not been clearly defined. Apoptosis is thought to play a key role in the mechanism, via caspase-dependent and caspase-independent pathways. Caspase-3, Apoptosis-inducing factor (AIF), and anti-apoptosis B-cell lymphoma 2 (Bcl-2) are hypothesized to be involved in PROM. Objective: To determine the role of caspase-dependent and caspase-independent pathways in the mechanism of PROM. Materials and Methods: This was a case-control study involving 42 pregnant women with gestational age between 20-42 wk. Participants were divided into the case group (with PROM) and control group (without PROM). Amniotic membranes were collected immediately after the delivery, and samples were taken from the site of membrane rupture. Immunohistochemical examination was done to determine the expression of Caspase-3, AIF, and Bcl-2. Results: The expressions of Caspase-3 (OR = 9.75; 95% CI = 2.16-43.95; p = 0.001) and AIF (OR = 6.60; 95% CI = 1.48-29.36; p = 0.009) were significantly increased, whereas, Bcl-2 expressions (OR = 8.00; 95% CI = 1.79-35.74; p = 0.004) were significantly decreased in the case group. Conclusion: High Caspase-3, AIF, and low Bcl-2 expression were the risk factors for PROM. Thus, it is evident that caspase-dependent and caspase-independent pathways are involved in the mechanism of PROM. Key words: Premature, Membrane, Apoptosis, Caspase, Pregnancy

CENTELLA ASIATICA EXTRACT INCREASED ON THE LEVEL OF INTERLEUKIN 6 (IL-6) IN MICE

Salmonellosis is still problem in many developing countries including Indonesia. Themain problem in controlling and handling the disease is that only few antibiotics are availableto cure the disease. In addition, the prolonged use of such antibiotics often lead to bacterialresistant against the antibiotics. A herbal drugs such as Centella asiatica (in Indonesia isknown as pegagan) contains triterphenoid saphonins which acts as immunostimulant capableof enhancing the phagocytic activity of macrophages. However, no study has been conductedto investigate the use pegagan in activating macrophage of mice infected with Salmonellatyphi. A study was therefore conducted to find out the ability of Cantella asiatica inenhancing on the level interluekine (IL)-6 following challenge with Salmonella typhi. It istherefore expected that herbal drug such as Cantella asiatica can be used as an alternativemedicine to prevent and cure salmonellosis in both animals and human.Experimental laboratory studies were conducted using Completely FactorialRandomized Design. Mice were divided into 4 groups and they were treated respectively withdestilated water (negative control), 125, 250, and 500 mg/kg bw of Centella asiatica extract.The treatment was conducted daily for 2 weeks and the mice were then inoculated with 105cells of S. typhi. The level of IL-6 response were examined 24 hours, 2 weeks, and 4 weeksafter inoculation with S. typhi.The result showed that treatment of mice with Centella asiatica extract significantly(p<0,05) enhaced IL-6 level of Balb/c mice following inoculation with S. typhi. The highes tIL-6 level were observed in mice treated Centella asiatica extract at the dose of 500 mg/kgBW (385,9257±125,4314 pg/ml serum). And the highest IL-6 level were observed at 2 weeksfollowing inoculation with S. typhi (533.4262 ± 81.7184 pg / ml).A further study is recomended to examine the celluler immune response and moredetailed study on the humoral immune response of animals or human before this herbal isused as alternatif medicine to prevent and cure typhoid fever. It is also important to study thebest preparation, the half life, and the side effect of Centella asiatica in human and animals

Antioxidant properties of topical Caulerpa sp. extract on UVB-induced photoaging in mice

Caulerpa sp., a genus of seaweed native to the Indo-Pacific region, has been known for its antioxidant properties and health benefits when consumed as food. Previous studies have reported Caulerpa sp.’s potential as a strong antioxidant, but its effects on the skin in a topical preparation, especially its role in UV protection, have not been studied extensively. Our study investigated the protective effects of 0.2% and 0.4% Caulerpa sp. extract gels on photoaging in the UVB-irradiated skin of Wistar mice. The subjects were divided into naive control, vehicle control, and 3 treatment groups (0.2% Caulerpa sp. extract gel, 0.4% Caulerpa sp. extract gel, and 0.02% astaxanthin gel as a standard antioxidant). The groups, except the naive control group, received a total of 840 mJ/cm2 of UVB irradiation in four weeks. Protective effects of the extract were measured through the evaluation of collagen expression, MMP-1 expression and levels, and 8-OhDG expression. Mice who received topical application of Caulerpa sp. extract gel had higher collagen expression, betterpreserved collagen structure, lower levels of MMP-1, and less MMP-1 and 8-OHdG expressions compared to the vehicle control group. There was no difference between different concentrations of the extract. Our findings demonstrated that topical application of Caulerpa sp. extract gel significantly protected UVB-irradiated mice skin from photoaging