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The Role of Interferon–Tau (Ifn–τ) in The Reproduction Handling of Female Ruminant Depamede, Sulaiman Ngongu
Indonesian Bulletin of Animal and Veterinary Sciences Vol 19, No 4 (2009)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (144.106 KB) | DOI: 10.14334/wartazoa.v19i4.912

Abstract

In ruminants, interferon-tau (IFN–τ)  is well known as an important pregnancy factor. Interferon-tau as antiluteolytic cytokine is secreted from the tropoblast during the time of implantation. About 10 – 15% of pregnancy losses in cows were possibly due to the lack of IFN–τ to maintain corpus luteum. Interferon-tau induces some early pregnancy factors which are expressed around 15 days post insemination. This has led that IFN–τ to be a strong candidate for the development of ruminant early pregnancy tests. The aim of this paper is to review the existing literatures on the roles of IFN–τ in biology reproduction as well as researches that have been carried out on reproductive biotechnology especially in regards to the development of early pregnancy test for ruminant. It can be concluded that administration of IFN–τ is able to improve reproductive performance of female ruminants experimentally. Interferon-tau and its derivates can be used to develop a rapid test for early pregnancy diagnostic.   Key words: Interferon-tau, early pregnancy test, ruminant
Assessment of the purity and characteristics of rat splenic T cells isolated by one-step discontinuous gradient of percoll Depamede, Sulaiman Ngongu Depamede
Indonesian Journal of Animal and Veterinary Sciences Vol 15, No 2 (2010)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (243.1 KB) | DOI: 10.14334/jitv.v15i2.653

Abstract

T cells isolated from spleen lymphocytes of rat (Rattus rattus) have been commonly used in an immunological research. However the T cells are still contaminated with other splenic cells.  Therefore a simple and reliable technique to isolate the T cells in one step process is needed. The aim of the present study was to assess whether T cells isolated from spleen lymphocytes of rat as a model by means of discontinuous density gradient of Percoll could give pure T cells with their activities as needed in an immunological research. Isolation was carried out in 5 density gradients of Percoll i.e. 1.052, 1.063, 1.075, 1.085, and 1.122 g/ml. The purity of the cell fractions was determined by immunocytochemistry and fluorescence-activated cell sorting (FACS) methods. The activity of each lymphocyte fraction was determined through cell proliferation assays by culturing the cells in the presence of phytohemaglutinin (PHA) and Concanavalin A (Con A) as T cells inducers. The FACS results show that fraction I and II were found to contain mostly B cells and macrophages. The fractions were obtained on the layers of gradient 1.052/1.063 and 1.063/1.075. Fraction III (gradient 1.075/1.085) contain T cells (43.2%) which consisted of CD4 (34.4%) and CD8 (18.6%), B cells (36.5%), and macrophages (10.0%). Fraction IV was strongly expressed T cell receptors (76.2%) which consisted of CD4 (63.6%) and CD 8 (20.0%), while B cells were found 20.0% and macrophages just about 0.6%. The activity of T cells as expressed by their respond to PHA and Con A shown that fraction III gave the most positive response, while fraction IV which contained the most ‘pure’ T cells gave less activity. From the present study it can be concluded that the discontinuous density gradient of Percoll method can be used to isolate and purify T cells in one step process. Furthermore, the present study indicated that T cells to proliferate in response to mitogen require other immune cells such as B cells and macrophages. Key words: Percoll, Lymphocyte, T Cell, Rat, Mitogen
Improvements of PCR Amplification of Guanine plus Cytosine-Rich Constructs of Mycobacterium tuberculosis Gene using DMSO Sabrina, Yunita; Lestarini, Ima Arum; Ekawanti, Ardiana; Sriasih, Made; Depamede, Sulaiman N.; Ali, Muhamad
Jurnal Kedokteran Vol 1 No 1 (2012)
Publisher : Faculty of Medicine Universitas Mataram

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Abstract

Vaccine research entered a new era when several useful molecular research tools were established. Instead of attenuated virulent microorganisms or killed virulent microorganisms, effective subunit vaccines were developed using recombinant DNA technology. By using the technology, selected genes of the virulent microorganisms can be amplified, cloned, expressed, and evaluated as vaccine components in challenge studies. However, a major bottleneck with the amplification of functional genes from Mycobacterium tubeculosis containing guanine plus cytosin-rich templates is often hampered by the formation of secondary structures like hairpins and higher melting temperatures. To solve this problem in this research, the amplification reaction was modified by addition of dimethyl sulfoxide (DMSO) into amplification reaction mixtures. It was found that 10% (v/v) of DMSO in the reaction mixture improved the amplification of GC-rich template of M. tuberculosis gene. This result indicating that amplification of unbalanced content of G and C deoxyribonucleotides genome could be improved using low-cost organic molecule, DMSO. Therefore, the DMSO should be widely useful as an enhancer to improve the amplification of GC rich construct from other genome. Keywords: Mycobacterium tubeculosis,vaccine, dimethyl sulfoxide, Guanine-Cytosine
PEMURNIAN ANTIBODI MONOKLONAL TERHADAP VIRUS HEPATITIS B YANG DIISOLASI DARI ASITES MENCIT BALB/C MENGGUNAKAN “HOMEMADE” PROTEIN A SPIN MINI KOLOM Ngongu Depamede, Sulaiman
Jurnal Kedokteran YARSI Vol 20, No 2 (2012): MEI - AGUSTUS 2012
Publisher : Lembaga Penelitian Universitas YARSI

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (133.615 KB) | DOI: 10.33476/jky.v20i2.160

Abstract

Penelitian ini bertujuan mengembangkan pemurnian antibodi monoklonal menggunakan eksmini kolom yang isinya diganti dengan partikel gelassilika yang terkonjugasi dengan Protein A. Matriks Protein A yang digunakan adalah Prosep vA, sedangkan antibodi target adalah antibodi monoklonal (MAb) terhadap antigen permukaan virus hepatitis B yang diisolasi dari asites mencit Balb/c. Sebelum pemurnian, sampel asites dibagi menjadi 3 yang masingmasing diinkubasikan dengan Protein A dalam 3 perlakuan. Perlakuan A dan B adalah inkubasi 30 dan 60 menit pada suhu kamar, sedangkan perlakuan C adalah inkubasi semalam pada suhu 4°C. Hasil pemurnian menunjukkan bahwa aktivitas spesifik dan tingkat kemurnian MAb secara berurutan adalah 358.45 U/mg, 237.83 U/mg untuk perlakuan C dan B, dan 229.67 U/mg untukperlakuan A. Hasil penelitian ini menunjukkan bahwa 'homemade' mini kolom yang dikembangkan dalam penelitian ini dapat digunakan untuk memurnikan MAb untuk kebutuhan riset skala laboratorium.Penelitian ini bertujuan mengembangkan pemurnian antibodi monoklonalmenggunakan eks-mini kolom yang isinya diganti dengan partikel gelassilikayang terkonjugasi dengan Protein A. Matriks Protein A yang digunakan adalahProsep vA, sedangkan antibodi target adalah antibodi monoklonal (MAb)terhadap antigen permukaan virus hepatitis B yang diisolasi dari asites mencitBalb/c. Sebelum pemurnian, sampel asites dibagi menjadi 3 yang masingmasingdiinkubasikandenganProteinAdalam3perlakuan.PerlakuanAdanBadalahinkubasi30dan60menitpadasuhukamar,sedangkanperlakuanCadalahinkubasisemalampadasuhu4°C.HasilpemurnianmenunjukkanbahwaaktivitasspesifikdantingkatkemurnianMAbsecaraberurutanadalah358.45U/mg,237.83 U/mg untuk perlakuan C dan B, dan 229.67 U/mg untukperlakuan A. Hasil penelitian ini menunjukkan bahwa 'homemade' mini kolomyang dikembangkan dalam penelitian ini dapat digunakan untuk memurnikanMAb untuk kebutuhan riset skala laboratorium.
Penghambatan Proliferasi Limfosit Mencit Balb/c oleh Ekstrak Testis Sapi Bali: Peran TGF-β S N Depamede; A Rosyidi
Media Peternakan Vol. 32 No. 2 (2009): Media Peternakan
Publisher : Faculty of Animal Science, Bogor Agricultural University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (314.299 KB)

Abstract

Testis extract (ET) of several animals such as rats, mice, boars, and bulls, has been reported able to suppress lymphocyte proliferation. In order to elucidate if ET of bali cattle has the suppressive activity, the present study was conducted. ET was isolated aseptically from the testis of bali cattle slaughtered at a slaughter house in Mataram, Lombok. After decapsulating, the testis were minced, homogenized (1 g tissue per 3 ml phosphate buffered saline, PBS, pH 7.2). The ET (40, 20, and 10µg/ml) was added on to lymphocytes isolated from Balb/c spleen, then cultured for 72 hours in RPMI media containing 10% foetal bovine serum in 96 well plates with the concentration of 250,000 cells per well, with- or without concanavalin A (10 µg/ml), or IL-2 (0.2 ng/ml). After 72 h of culture, cell proliferation was examined by colorimetric assay. The results showed that bali cattle ET were able to suppress the proliferation of lymphocytes in vitro through the process of apoptosis. Western blot analysis revealed that bali cattle ET contain TGF-β1. The suppression effects of the ET were eliminated by anti-TGF-β1 antibody. It can be concluded that ET of bali cattle expressed TGF-β  which inhibited lymphocyte proliferation through an apoptotic cell mechanism. The actual roles of TGF-β  in the bali cattle testis need to be elucidated further. Key words: bali cattle testis, lymphocyte, proliferation, TGF-β1, apoptosis
Development of a Rapid Immunodiagnostic Test for Pork Components in Raw Beef and Chicken Meats: a Preliminary Study S N Depamede
Media Peternakan Vol. 34 No. 2 (2011): Media Peternakan
Publisher : Faculty of Animal Science, Bogor Agricultural University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1102.898 KB) | DOI: 10.5398/medpet.2011.34.2.83

Abstract

A rapid immunodiagnostic test that provides visual evidence of the presence of pork components in raw beef and chicken meats was developed. Colloidal gold was prepared and conjugated with anti-Swine IgG polyclonal antibody. Immunochromatographic test strips were produced, and then were used to test laboratory adulterated raw meat samples. The samples consisted of pork-in-beef, or pork-in-chicken at 1/0; 1/100; 1/1,000; 1/5,000; 1/10,000 (w/w) adulteration levels that were extracted in phosphate-buffered saline. Raw beef and chicken meats without pork were included as controls. Analysis was completed in 10 min. Detection limit was 1/5,000 (w/w), although 1/10,000 was also observed. This immunodiagnostic tests can be conveniently applied to detect low levels of pork components in raw beef and chicken meat products. For the commercial purposes, further studies need to be carried out.
Identifikasi Mutasi FecX Pada Gen BMP15 dan Pengaruhnya Terhadap Sifat Prolifik pada Kambing Lokal di Kabupaten Lombok Barat Rahmat Agus Hidayat; Sulaiman Ngongu Depamede; Maskur Maskur
Jurnal Ilmu Dan Teknologi Peternakan Indonesia (JITPI) Indonesian Journal of Animal Science and Technology) Vol 1 No 1 (2015): Jurnal Ilmu dan Teknologi Peternakan Indonesia (JITPI)
Publisher : Faculty of Animal Husbandry, University of Mataram

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (236.638 KB) | DOI: 10.29303/jitpi.v1i1.3

Abstract

The aims of this study were to identify the mutations of FecX gene in the local goats and to analyze its polymorphism as well as its influence on the prolific nature of the local goats in West Lombok Regency, Indonesia. The study was conducted in the Immunobiology Laboratory of Mataram University, using DNA isolated from 100 blood samples of local female goats which have given birth of once to three times. The methods used were PCR-RFLP method and the PCR products were digested with HinfI restriction enzyme (G|ANTC) then analyzed visually based on DNA banding patterns on 2% agarose gels. The frequency of allele and genotype obtained, were then analyzed through a comparison with the secondary data of litter size obtained from the local goat keepers information. The results showed that the gene mutation of FecXG produced two alleles: "wild-type" (+) sized of 110 bp and 31 bp, and the mutant allele (G) of 141 bp with the allele frequency of 0,965 and 0,035 respectively. Combinations of alleles in the gene BMP15 produced two genotypes, namely (a) genotype ++ (110 bp/110 bp) with a frequency of 0.93, with the average litter size of 1.59 ± 0.319, and (b) genotype G + (141bp/110 bp), with a frequency of 0.07 and with the average litter size of 1.65 ± 0.202. The results of this study indicated that mutation occurred in BMP15 gene, i.e. FecXG gene, the gene responsible for the prolificacy of animals studied. Furthermore there was a correlation between polymorphism of FecXG gene and the prolific nature of the local goats, which was predicted to lead the divergence in litter size of each local goat genotype
Perubahan Status Fisiologis dan Bobot Badan Sapi Bali Bibit yang Diantarpulaukan dari Pulau Lombok ke Kalimantan Barat Ambius Anton; Lalu Muhammad Kasip; Lalu Wira Pribadi; Sulaiman Ngongu Depamede; A. Rai Somaning Asih
Jurnal Ilmu Dan Teknologi Peternakan Indonesia (JITPI) Indonesian Journal of Animal Science and Technology) Vol 2 No 1 (2016): Jurnal Ilmu dan Teknologi Peternakan Indonesia (JITPI)
Publisher : Faculty of Animal Husbandry, University of Mataram

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (146.521 KB) | DOI: 10.29303/jitpi.v2i1.17

Abstract

The aims of this study were to investigate the changes of physiology and body weight of Bali cattle heiferstransported from Lombok Island to West Kalimantan. This was a case study on effect of transportation by shipof Bali cattle heifers from the Lombok Island to West Kalimantan. Twenty Bali cattle heifers with the initialbody weight of 123.28±11.00 kg were observed in this study. Samples were determined using the purposivesampling of the total 378 cattle during transportation. Body temperature, respiratory, pulse, hemoglobin,hematocrit, blood glucose and body weight of the heifers were measured prior to transportation (T0) and aftertransportation (T1). The temperature and humidity of the environment in the ship during the transportationwere also noted. Data were analyzed by using descriptive analysis (Arithmetic means) and the differenceswere tested using T-Test. The results show that livestock transportation had some negative effects on theperformances of the heifers. The average of the ship paddock temperature and the humidity during thetransportation period were 28.83±0.860C and 72.57±4.08%, respectively. The physiological status of bodytemperature; pulse; respiratory and blood glucose concentrations increased (P>0.05) by 0.600C; 8.25time/min; 7.6 times/min and 8.7 mg/dL, respectively. Whilst, the hematocrit levels, hemoglobin concentrationand body weight of the heifers decreased significantly (P>0.05) by 2.73%; 0.90 g/dL; and 21.23 kg/head,respectively. The high decrease of the heifers’ body weight changes (17.22%) was suspected to be due to lackof feed and water intakes during transportation. It is suggested that the heifers should have enough feed andwater consumption during tranportation for reducing stress.
Infection and Distribution of Liver Fluke (Fasciola Sp) Zoonotic Parasite on Cattle in Central Lombok and East Lombok Districts Wayan Wariata; Made Sriasih; Anwar Rosyidi; Muhamad Ali; Sulaiman N. Depamede
Jurnal Ilmu Dan Teknologi Peternakan Indonesia (JITPI) Indonesian Journal of Animal Science and Technology) Vol 5 No 2 (2019): Jurnal Ilmu dan Teknologi Peternakan Indonesia (JITPI)
Publisher : Faculty of Animal Husbandry, University of Mataram

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (174.317 KB) | DOI: 10.29303/jitpi.v5i1.55

Abstract

Salah satu faktor yang sangat mempengaruhi populasi dan produksi ternak adalah masalah penyakit dan parasit. Gejala serangan parasit seringkali tidak tampak dan kadang-kadang petani kurang menyadarinya. Cacing hati (Fasciola sp.) merupakan parasit zoonosis yang dapat menginfeksi berbagai macam hewan ternak ruminansia terutama sapi. Fasciolamemiliki dua spesies utama yaitu hepatica dan gigantica. Penyebaran Fasciola sp. adalah daerah beriklim tropis basah yaitu di Asia dan Afrika. Fasciola sp. yang sering dijumpai di Indonesia adalah spesies gigantica, sedangkan spesies hepatica sering terdeteksi pada sapi-sapi impor. Untuk mengetahui infeksidan tingkat penyebaran Fasciola giganticadan Fasciola hepaticapada ternak sapi di kabupaten Lombok Tengahdan Lombok Timurmaka dilakukan survei, pengamatan dan pengambilan sampel organ hati sapi di pasar-pasar daging yang ada pada dua daerah tersebut. Pada masing-masing daerah diambil 3 lokasi pasar daging, yaitu di kabupaten Lombok Tengah: pasar Praya, pasar Mantangdan pasar Kopang; di kabupaten Lombok Timur: pasarSelong, pasarMasbagikdan pasarTerara. Pada masing-masing pasar, pengamatan dan pengambilan sampel diulang sebanyak 3 kalipada 3 orang pedagang.Selanjutnya sampel diamati di laboratorium untuk mengetahui adanya cacing hati dan kerusakan hati sapi. Hasil penelitian menunjukkan bahwa, sapi-sapi dan daging sapi yang diperjual belikan di pasar-pasar Lombok Tengah dan Lombok Timur terinfeksi parasit cacing hati (Fasciola sp.), dengan tingkat infeksi yang masih rendah, yaitu antara 1,3 sampai 2,3. Di samping itu, warna dan tekstur hati sapi yang diperjual belikan di pasar-pasar Lombok Tengah dan Lombok Timur relatif masih baik dan layak dikonsumsi.
Assessment of the purity and characteristics of rat splenic T cells isolated by one-step discontinuous gradient of percoll Sulaiman Ngongu Depamede Depamede
Jurnal Ilmu Ternak dan Veteriner Vol 15, No 2 (2010): JUNE 2010
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (243.1 KB) | DOI: 10.14334/jitv.v15i2.653

Abstract

T cells isolated from spleen lymphocytes of rat (Rattus rattus) have been commonly used in an immunological research. However the T cells are still contaminated with other splenic cells.  Therefore a simple and reliable technique to isolate the T cells in one step process is needed. The aim of the present study was to assess whether T cells isolated from spleen lymphocytes of rat as a model by means of discontinuous density gradient of Percoll could give pure T cells with their activities as needed in an immunological research. Isolation was carried out in 5 density gradients of Percoll i.e. 1.052, 1.063, 1.075, 1.085, and 1.122 g/ml. The purity of the cell fractions was determined by immunocytochemistry and fluorescence-activated cell sorting (FACS) methods. The activity of each lymphocyte fraction was determined through cell proliferation assays by culturing the cells in the presence of phytohemaglutinin (PHA) and Concanavalin A (Con A) as T cells inducers. The FACS results show that fraction I and II were found to contain mostly B cells and macrophages. The fractions were obtained on the layers of gradient 1.052/1.063 and 1.063/1.075. Fraction III (gradient 1.075/1.085) contain T cells (43.2%) which consisted of CD4 (34.4%) and CD8 (18.6%), B cells (36.5%), and macrophages (10.0%). Fraction IV was strongly expressed T cell receptors (76.2%) which consisted of CD4 (63.6%) and CD 8 (20.0%), while B cells were found 20.0% and macrophages just about 0.6%. The activity of T cells as expressed by their respond to PHA and Con A shown that fraction III gave the most positive response, while fraction IV which contained the most ‘pure’ T cells gave less activity. From the present study it can be concluded that the discontinuous density gradient of Percoll method can be used to isolate and purify T cells in one step process. Furthermore, the present study indicated that T cells to proliferate in response to mitogen require other immune cells such as B cells and macrophages. Key words: Percoll, Lymphocyte, T Cell, Rat, Mitogen