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All Journal Jurnal Sain Veteriner Indonesian Journal of Biotechnology Journal of Health (JoH) CHEESA: Chemical Engineering Research Articles
Asmarani Kusumawati
Fakultas Kedokteran Hewan, Universitas Udayana, Bali
Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Health Professions Immunology & microbiology Materials Science & Nanotechnology Medicine & Pharmacology Nursing Public Health Veterinary

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Characterization of envelope-transmembrane Gene of Jembrana Disease Virus Tabanan 1995 Isolate Kusumawati, Asmarani; Pratiwi, Rarastoeti; Astuti, Pudji; Hamid, Penny Humaidah
Indonesian Journal of Biotechnology Vol 15, No 1 (2010)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (299.935 KB)

Abstract

The availability of specific and rapid detection methods is essential for monitoring the health status of farmed species, particularly in viral disease as in this case early diagnosis is a critical factor in containing disease outbreaks. Jembrana Disease Virus (JDV) is a lentivirus that causes an acute, severe disease syndrome in infected Bali cattle in Indonesia, resulting in heavy economic losses because of the high mortalities. The virus-host interaction and the modes of transmission are still unknown. The goal of the research was to designa probe candidate of Jembrana Disease Virus based on envelope-transmembrane (env-tm) gene to optimize Jembrana disease detection method. The DNA fragment derived from env-tm of JDV was used, cloned in pGEX-TM and expressed in E.coli DH 5α. Sequence analysis was conducted with BLAST programs from NCBI. Sequence analyses of the fragments of env-tm clone, indicated that it has a very closed genetic relation with 97,68% homology identity. Probe was designed based on the conserved region of env-tm using Geneious resulted in JT2 252 bp long. BLAST analyses showed that probes had high specifity to other strains of JDV in Indonesia.Key words : probe, env-tm, JDV, specifity, sensitivity.
DIGoxigenin (DIG) Labeled Probe Candidate of Surface Antigen 1 (SAG1) for Toxoplasma gondii Detection Kusumawati, Asmarani; Septiana, Nafratilova; Hartati, Sri
Indonesian Journal of Biotechnology Vol 16, No 1 (2011)
Publisher : Universitas Gadjah Mada

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Abstract

Toxoplasma gondii is one of the opportunistic pathogen that causes toxoplasmosis. Infection of Toxoplasma gondii has been estimated as high both in human and animal. The manifestation of infection were abortion, hydrocephalus, brain calcification, chorioretinal scar, and loss of productivity even to death in patients with acquired immunosuppression. Early diagnostic method which are rapid and accurate is essential for T. gondii detection because of its high prevalence. The purpose of this study was to develop a sensitive probes derived from Surface Antigen 1 (SAG1) for detection T. gondii and to examine the specificity and sensitivity of probe as diagnostic tool for toxoplasmosis. This research used SAG1 gene of T. gondii local isolate IS-1 that was cloned into pGEX-2T and transformed into Eschericia coli DH5α. The sequence of SAG1 was labeled with DIGoxigenin (non radioactive labeled) using PCR DIG Labeling Mix to derive 213 bp (probe-TS). BLAST and dot-blot hybridization analyses showed that probes had high specifity with other strains of T. gondii. Probe was able to detect T. gondii DNAup to 10 ng/μl of total sample DNA.
Pengaruh Ukuran Ovarium dan Folikel terhadap Penampilan Reproduksi Pada Sapi PO dan SimPO di Kecamatan Jatinom, Kabupaten Klaten Priyo Jr, Topas Wicaksono; Budiyanto, Agung; Kusumawati, Asmarani
Jurnal Sain Veteriner Vol 38, No 1 (2020): April
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (423.167 KB) | DOI: 10.22146/jsv.57362

Abstract

The increasing beef cattle population in Indonesia is not significantly with high demand of meat every year. The insignificant increasing in population is caused by fertility decreasing, infectious disease and the reduction amount of forage land. The factor which causes cattle fertility decreasing can’t be separated from the ovarian and follicular diameter which have an effect on high various reproductive appearance. Parameters that used for knowing reproductive appearance of cattle are Service per Conception (S/C) and Calving Interval (CI). The aims of this study was to describe ovary sizes and follicular diameter in cattle, to describe ovary and follicular diameter toward S/C and CI variation. This research uses female cattle, 8 Simmental-PO (SimPO) and 9 Peranakan Ongole breeds (PO), multiparous, are not pregnant, 4-8 years old, Body Condition Score (BCS) 2.5-3.5, peak phase estrus, has no reproductive problems, clear recordings and live in the area of farmer in Jatinom District, Klaten Regency, Central Java Province. Cattle are examined for ovarian size and follicle size using ultrasonography. S/C and CI data were obtained from records of artificial insemination cards (AI). The data obtained were analyzed by t test. The results of this study showed there were no differences in the diameter of ovaries in the size of SimPO and PO cattle (P> 0.05), there were no differences in the diameter of SimPO and PO cattle follicles (> 0.05), there were differences in ovarian and follicular diameter sizes with respect to S/C and CI in cattle with good and bad reproductive performance (P <0.05). The conclusion of the study showed that there was no difference in ovarian and follicular diameter size in SimPO and PO cattle (P> 0.05) but there was an influence on S/C and CI (P <0.05). 
Optimasi Kodon dan Konstruksi Plasmid Rekombinan Protein RBD SARS-CoV-2 pada E. coli BL21(DE3): Codon Optimization and Construction of Recombinant Plasmid for SARS-CoV-2 RBD Protein on E. coli BL21(DE3) Purnamasari, Elly Widyarni Eka; Dini Wahyu Kartika Sari; Tenri Ashari Wanahari; Khairun Nissa; Kusumawati, Asmarani
Journal of Health (JoH) Vol 11 No 1 (2024): Journal of Health (JoH) - January
Publisher : LPPM STIKES Guna Bangsa

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.30590/joh.v11n1.715

Abstract

Coronavirus Disease 2019 (COVID-19), caused by the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) resulted over 7 million deaths worldwide. Comprehensive strategies are necessary to combat this pandemic, especially vaccination to mitigate further infection and devastation. The development of vaccines hinges upon the utilization of immunogenic proteins from the SARS-CoV-2 virus, with particular emphasis on the receptor-binding domain (RBD) located within the spike glycoprotein’s subunit 1. This protein plays a key role in the virus’s interaction with host cells via ACE2 receptor to infect the host along with the subunit 2 to mediate the cell membrane fusion. While there are multiple vaccines in existence, there is a requirement to create more affordable and accessible vaccine alternatives, especially in developing countries. Potential improvements can be implemented within the vaccine development paradigm, including the exploration of recombinant protein-based vaccines as a viable alternative. In the present study, we optimized the RBD gene sequence as the critical step to construct a recombinant plasmid for heterologous prokaryotic protein expression in Escherichia coli BL21(DE3). The optimized codon then inserted into the plasmid vector pET-15b and constructed a recombinant plasmid for transformation into cell host by heat-shock method. The study was evaluated and validated through colony PCR, plasmid isolation, and sequencing. These findings offering a foundational basis for further research in the expression and purification of RBD SARS-CoV-2 protein in E. coli BL21(DE3) as a candidate for subunit recombinant protein vaccine or diagnostic tool in COVID-19 research.
Chitosan Potential as Delivery Agent for S1 Gene from SARS-CoV-2 with pEGFP-N1 as its Vectors: Potensi Kitosan sebagai Sistem Penghantaran Gen S1 Virus SARS-CoV-2 dengan Vektor pEGFP-N1 Kusumawati, Asmarani; Fahhlevi, Nabilah Adzra; Pratiwi, Rarastoeti; Jannah, Raudatul; Maulina, Nurina Tahta Afwi
Journal of Health (JoH) Vol 11 No 02 (2024): Journal of Health (JoH) - July
Publisher : LPPM STIKES Guna Bangsa

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.30590/joh.v11n2.716

Abstract

Coronavirus Disease-19 (COVID-19) is a disease caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), this disease began to emerge at the end of 2019 and has become a pandemic to this day. Vaccination can play an important role to prevent the spread of the disease, so there are many vaccine developments, including DNA vaccines. Delivery systems such as chitosan can increase the effectiveness of vaccines. The purpose of this study was to optimize and evaluate the pEGFP-N1-S1 formula with a chitosan delivery system as a DNA vaccine candidate. Plasmids were cloned in E.coli DH5α and extracted for further evaluation with restriction enzymes and sequencing. The EGFP-N1-S1 plasmid was then formulated with chitosan nanoparticles with a mass ratio of Plasmid DNA:chitosan of 1:0.1; 1:0.2; 1:0.3; 1:0.4; 1:0.5; 1:0.6 and 1:0.7. The isolated plasmid has a concentration of 2182.32 ng/L and a purity of 1.911. Restriction results showed that the S1 gene (684 bp) was successfully inserted into the pEGFP-N1 plasmid (4813 bp). DNA:chitosan mass ratio of 1:0.6 is the optimal formulation for DNA to bind with chitosan perfectly.
Efek Penambahan Antioksidan Selenium, Kurkumin dan Kombinasinya Terhadap Motilitas, Recovery Rate dan Viabilitas Spermatozoa pada Kriopreservasi Semen Sapi Peranakan Ongole Yuniar, Resa Miftahatu; Kusumawati, Asmarani; Setyawan, Erif Maha Nugraha
Jurnal Sain Veteriner Vol 42, No 3 (2024): Desember
Publisher : Faculty of Veterinary Medicine, Universitas Gadjah Mada bekerjasama dengan PB PDHI

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jsv.85255

Abstract

Salah satu kendala dalam pelaksanaan kriopreservasi semen adalah tingginya kadar reactive oxygen species (ROS) yang dapat menyebabkan reduksi pada motilitas sperma dan viabilitas sperma post thawing. Strategi untuk mengatasi tingginya kadar ROS adalah dengan menambahkan antioksidan pada pengencer selama proses kriopreservasi. Tujuan dari penelitian ini adalah mengetahui pengaruh penambahan antioksidan selenium dan kurkumin dalam menurunkan stress oksidatif pada proses kriopreservasi semen sapi peranakan ongole (PO). Penelitian ini menggunakan semen yang dikoleksi dari sapi PO dengan vagina buatan. Sampel semen yang ditampung diperiksa secara mikroskopis dan makroskopis kemudian dibagi menjadi 4 perlakuan yaitu kontrol, penambahan selenium (sodium selenite) dengan konsentrasi 50 µM, kurkumin 10 µM dan kombinasi antara selenium 50 µM dengan kurkumin 10 µM.  Semen yang telah dicampur dengan pengencer dan antioksidan kemudian dibekukan dalam proses kriopreservasi, selanjutnya parameter yang diamati meliputi pemeriksaan motilitas sperma, recovery rate dan viabilitas sperma. Semua parameter dianalisis statistika menggunakan One Way ANNOVA dan uji DMRT. Berdasarkan penelitian menunjukkan bahwa perlakuan kurkumin 10 µM memiliki nilai motilitas 48.00 ± 4.47%, recovery rate 66.66 ± 5.54% dan viabilitas 67.00 ± 4.30%, sedangkan perlakuan selenium 50 µM memiliki nilai presentase motilitas 46,00 ± 4,18%, recovery rate 63.90 ± 5.59% dan viabilitas 64.00 ± 3.93%. Pemanfaatan kombinasi kurkumin dan selenium tidak memberikan pengaruh yang nyata dibandingkan dengan kontrol. Kesimpulan penelitian ini adalah penambahan antioksidan kurkumin dan selenium bermanfaat dalam mempertahankan kualitas sperma sapi PO, namun perlakuan kombinasi kurang baik dalam mempertahankan kualitas sperma.
Deteksi dan Identifikasi Senyawa Flavonoid Ekstrak Etanol Kulit Batang Tanaman Majapahit (Crescentia cujete) dengan LCMS Fatimah, Fatimah; Martha, Rahma Diyan; Kusumawati, Asmarani
CHEESA: Chemical Engineering Research Articles Vol. 3 No. 2 (2020)
Publisher : Universitas PGRI Madiun

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.25273/cheesa.v3i2.7688.88-98

Abstract

Tanaman Majapahit (Crescentia cujete) merupakan salah satu tanaman yang banyak tersebar di Indonesia. Namun, tanaman ini kurang mendapatkan perhatian, karena kurangnya informasi mengenai potensi tanaman tersebut. Flavonoid merupakan salah satu kelompok senyawa terbesar yang terdapat pada tanaman yang memiliki beberapa fungsi farmakologis dan medisinal, salah satunya anti kanker. Dalam penelitian ini, dilakukan deteksi dan identifikasi senyawa flavonoid pada ekstrak etanol kulit batang tanaman majapahit menggunakan LCMS, untuk mengetahui potensi tanaman majapahit sebagai salah satu kandidat tanaman potensial antikanker. Berdasarkan hasil deteksi menggunakan LCMS diketahui terdapat sekitar 88 senyawa yang terdeteksi, termasuk didalamnya 12 senyawa flavonoid. Senyawa flavonoid yang ditemukan, yaitu; acetoin, quercetine, kaempferol-3-O-rhamnoside, acacetin7-rutinoside, fortunellin, kaempferol 3-[6"-(3-hydroxy-3-methylglutaryl) glucoside], didymin, diosmin, hesperidin, rutin, narirutin 4'-glucoside, kaempferol 3-[6"-(3-hydroxy-3-methylglutaryl) glucoside]-7-glucoside. Kaempferol-3 -O-rhamnoside merupakan flavonoid dengan komposisi terbesar yakni, sebesar 4,072%. Keseluruhan flavonoid yang teridentifikasi memiliki potensi sebagai antikanker, kecuali acetoin dan fortunellin.
Co-Authors A.A. Ketut Agung Cahyawan W Agung Budiyanto Dini Wahyu Kartika Sari Erif Maha Nugraha Setyawan Fahhlevi, Nabilah Adzra fatimah Fatimah Khairun Nissa Maulina, Nurina Tahta Afwi Nafratilova Septiana, Nafratilova Penny Humaidah Hamid, Penny Humaidah Priyo Jr, Topas Wicaksono Purnamasari, Elly Widyarni Eka Rahma Diyan Martha Rarastoeti Pratiwi raudatul jannah Sri Hartati Tenri Ashari Wanahari Yuniar, Resa Miftahatu