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Expression of SARS-CoV-2 Nucleocapsid (N) Recombinant Protein Using Escherichia coli System Rizki Aulia Ansari; Uus Saepuloh; Silmi Mariya; Yuliana; Rachmitasari Noviana; Irma Herawati Suparto; Huda Shalahudin Darusman
HAYATI Journal of Biosciences Vol. 30 No. 3 (2023): May 2023
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.30.3.445-450

Abstract

One of the main antigen that can be used for serological testing is the nucleocapsid (N) which is the most abundant viral-derived protein in SARS-CoV-2 where this virus can cause COVID19 disease. The aim of this study was to develop the SARS-CoV-2 N recombinant protein using Escherichia coli expression system. A total of 1,089 nucleotides encoding 362 amino acids of SARS-CoV-2 N was cloned to pET-14b vector. The plasmid then expressed in E. coli BL21 (DE3) and induced with 1.0 mM IPTG (Isopropyl-β-d-1-thiogalactopyranoside). The cell was harvested using denaturation lysis buffer due to inclusion body formation of SARS-CoV-2 N protein. Dialysis processed and concentrated using PEG-6000 resulted 0.992 mg/ml protein yield. Analysis of SARS-CoV-2 N recombinant protein using SDS-PAGE technique showed approximately 37.0 kDa specific band target protein. Application of this SARS-CoV-2 N recombinant protein to vaccinated and non-vaccinated antibody serum samples using ELISA technique indicated the significant result of optical density mean at 0.603 and 0.135, respectively. This study revealed that the production of SARS-COV-2 N recombinant protein could be carried out in E. coli expression system under denatured conditions, therefore the methods are more effective in producing the protein as a basic material in immuno-diagnostic assay.
MONYET EKOR PANJANG (Macaca fascicularis) SEBAGAI HEWAN MODEL DAN ETIKA PENGGUNAANNYA Ratni Ernita; Tatang Mitra Setia; Fitriya Nur Annisa Dewi; Huda Shalahudin Darusman
Gunung Djati Conference Series Vol. 6 (2021): Seminar Nasional Biologi (SEMABIO) 6 Tahun 2021
Publisher : UIN Sunan Gunung Djati Bandung

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (376.278 KB)

Abstract

The use of nonhuman primates as animal models in biomedical research has an important role in the development of human health. A close phylogenetic relationship with humans as well as similarities in genetics, behavior and biochemical activities are some of the things that underlie the selection of nonhuman primates as animal models. Cynomolgus macaques (Macaca fascicularis) is one of Indonesia's endemic primates which is widely used in research. Its current status as Appendix II in Convention International Trade in Endangered Species of Wild Fauna and Flora (CITES) defines that the population remains quite large in nature, which allows this species to be used in research, as long as they are sourced from the captive-bred population. Although the use of nonhuman primates in research is unavoidable, it is warranted that potential harm such as distress, discomfort, stress, pain and even death have to be avoided or minimized. Implementation of ethics is therefore required for the care and use of cynomolgus macaques in research, such as by assuring that the 3R principles (replacement, reduction, refinement) are met. Refinement can be achieved by applying the principles of animal welfare known as the Five Freedoms of Animal Welfare (5F). The active implementation of these ethical principles continues to develop along with the development of science and technology, and receives more spotlight in the world of biomedical research globally. Ensuring adequate implementation of ethical principles in research with nonhuman primates in each research institution must be under the oversight of an IACUC (Institutional Animal Care and Use Committee), which will weigh on the harm to animals compared to the benefits to humans. Active implementation of the principles of ethics and animal welfare will result in increasing the quality of research, serves as an assurance to the society, and in compliance to the requirements for good scientific publication.
Analyses of Vegetation Used by Long-tailed Macaque (Macaca fascicularis Raffles 1821) in Tinjil Island Dyah Perwitasari-Farajallah; Hana Intishar Sawitri; Silvy Thiyana; Tommy Langgeng Abimanyu; Entang Iskandar; Huda Shalahudin Darusman
Journal of Tropical Biodiversity and Biotechnology Vol 8, No 1 (2023): April
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jtbb.70739

Abstract

Tinjil Island is a semi-natural breeding facility for long-tailed macaques (Macaca fascicularis) managed by Primate Research Center, IPB University, located at the southern of Java Island and surrounded by the Indian Ocean. Long-tailed macaques are considered frugivorous even though they are well-known for their flexible diet. This study aims to analyse the vegetation supporting the population of long-tailed macaques. Data were collected from six tracks using square sampling plots with the size of 20 m x 20 m for trees as the main plot, inside the main plot were square subplots consisting of 10 m x 10 m for poles, 5 m x 5 m for saplings, and 2 m x 2 m for seedlings. The Important Value Index (IVI) was calculated for each level of vegetation. Hanjuang (Dracaena elliptica) dominated the seedlings with 29.35%, followed by Kampis (Hernandia peltata) with 18.73%, and Kalapari (Pongamia pinnata) with 13.73%. Hanjuang (Dracaena elliptica) also dominated the saplings with 26.83%, followed by Pancal (Syzygium antisepticum) with 19.19%, and Laban (Vitex pubescens) with 12.30%. The poles were dominated by Ki Cau (Dolichandrone spathacea) as high as 59.28%, while Waru (Thespesia populnea) and Ki Ciat (Ficus septica) dominated at 40.47% and 36.15%, respectively. Kampis (Hernandia peltata) dominated the trees with 39.28%, followed by Ki Ara (Ficus glomerata) with 35.56%, and Ki Langir (Dysoxylum amooroides) with 28.70%. Species found on Tinjil Island are mostly Moraceae (9.84%) and Fabaceae (9.84%), followed by Malvaceae (8.20%), Euphorbiaceae (4.92%), Myrtaceae (4.92%), and Anacardiaceae (4.92%). The vegetation in Tinjil Island supports the livelihood of long-tailed macaques on the island because they have an abundance of food and staple food such as figs to help them fulfil the energy needed to survive and reproduce. 
CHARACTERIZATION OF MESENCHYMAL STEM CELLS FROM WHITE ADIPOSE TISSUE OF Macaca fascicularis Harrold Halilintar MR Subu Taopan; Fitriya Nur Annisa Dewi; Silmi Mariya; Huda Darusman; Permanawati Permanawati; Dondin Sajuthi; Irma Herawati Suparto
Jurnal Kedokteran Hewan Vol 15, No 4 (2021): December
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (526.996 KB) | DOI: 10.21157/j.ked.hewan.v15i4.18203

Abstract

 Mesenchymal stem cells (MSC) derived from white adipose tissue are considered to be an appropriate therapeutic candidate for various human diseases due to their natural ability to maintain homeostasis. They have also been considered for use as a therapeutic alternative due to their sufficient availability and ease of harvesting. MSC play an important role in the process of adipogenesis and energy metabolism that is directly related to the pathogenesis of various degenerative diseases and metabolic syndromes. The purpose of this study was to obtain MSC cultures from white adipose tissue biopsied from three adult males Macaca fascicularis and to evaluate these cultures qualitatively and quantitatively regarding their characteristics and ability to differentiate. Polymerase chain reaction (PCR) was used to confirm the expression of mesenchymal stem cell-specific gene markers, and a specific growth medium was used to differentiate these cells into adipocytes, chondrocytes, and osteocytes. The results obtained showed that the MSC culture from M. fascicularis WAT had a fibroblast-like morphology with a spindle shape. The MSC markers of CD73, CD90 and CD105, were also expressed. The MSC culture was also able to differentiate into adipocytes, chondrocytes, and osteocytes. Based on these results, white adipose tissue from Macaca fascicularis was found to have potential as a source of MSC.
CHARACTERIZATION OF MESENCHYMAL STEM CELLS FROM WHITE ADIPOSE TISSUE OF Macaca fascicularis Harrold Halilintar MR Subu Taopan; Fitriya Nur Annisa Dewi; Silmi Mariya; Huda Darusman; Permanawati Permanawati; Dondin Sajuthi; Irma Herawati Suparto
Jurnal Kedokteran Hewan Vol 15, No 4 (2021): December
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21157/j.ked.hewan.v15i4.18203

Abstract

 Mesenchymal stem cells (MSC) derived from white adipose tissue are considered to be an appropriate therapeutic candidate for various human diseases due to their natural ability to maintain homeostasis. They have also been considered for use as a therapeutic alternative due to their sufficient availability and ease of harvesting. MSC play an important role in the process of adipogenesis and energy metabolism that is directly related to the pathogenesis of various degenerative diseases and metabolic syndromes. The purpose of this study was to obtain MSC cultures from white adipose tissue biopsied from three adult males Macaca fascicularis and to evaluate these cultures qualitatively and quantitatively regarding their characteristics and ability to differentiate. Polymerase chain reaction (PCR) was used to confirm the expression of mesenchymal stem cell-specific gene markers, and a specific growth medium was used to differentiate these cells into adipocytes, chondrocytes, and osteocytes. The results obtained showed that the MSC culture from M. fascicularis WAT had a fibroblast-like morphology with a spindle shape. The MSC markers of CD73, CD90 and CD105, were also expressed. The MSC culture was also able to differentiate into adipocytes, chondrocytes, and osteocytes. Based on these results, white adipose tissue from Macaca fascicularis was found to have potential as a source of MSC.
THE GENE EXPRESSION OF ADAM17 AS A GENETIC MARKER OF ALZHEIMER DISEASES IN THE BRAIN OF LONG-TAILED MACAQUES (Macaca fascicularis) Lis Rosmanah; Karin katina Hasya; Uus Saepuloh; Wasmen Manalu; Adi Winarto; Etih Sudarnika; Huda Shalahudin Darusman
Jurnal Kedokteran Hewan Vol 17, No 3 (2023): September
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21157/j.ked.hewan.v17i3.31014

Abstract

This study aims to identify the expression of the a disintegrin and metalloproteinase 17 (ADAM17) gene as a marker of Alzheimer's disease in long-tailed macaques (Macaca fascicularis). This study used six brain samples (hippocampus and cortex regions) of long-tailed macaques which was divided into two groups consisting of aged long-tailed macaques and adult long-tailed macaques. The expression of ADAM17 gene was determined by comparing the relative quantification values between the two age groups, and brain regions consisting of the hippocampus and cortex regions. The results of data analysis showed no significant difference in the expression of ADAM17 gene between brain regions and between age groups of long-tailed macaques. However, numerically the results showed a lower expression of ADAM17 gene in the hippocampus region of aged macaques. Lower expression of ADAM17 gene could be a marker of old animals indicating the stages of Alzheimer's disease.
Expression of APP, CDK5, and AKT1 Gene Related to Alzheimer Disease in Brain of Long-tailed Macaques Lis Rosmanah; Uus Saepuloh; Sela Septima Mariya; Irma Herawati Suparto; Wasmen Manalu; Adi Winarto; Huda Shalahudin Darusman
HAYATI Journal of Biosciences Vol. 31 No. 1 (2024): January 2024
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.31.1.145-152

Abstract

Amyloid plaques and Neurofibrillary Tangles (NFTs) are known to be key pathological features of Alzheimer disease. To gain a better understanding of this disease, studies were carried out on the Indonesian primates, the long-tailed macaques, using a spontaneous Alzheimer's disease model. Examining and identifying genetic markers involved in plaque formation and NFTs in long-tailed macaques is necessary to reveal their physiological processes. In this study, the expression of genes involved in the development of amyloid plaque (Amyloid Precursor Protein (APP)) and those that control the phosphorylation of tau protein (CDK5 and AKT1) was examined in the long-tailed macaque brain. This study showed that APP, CDK5, and AKT1 may potentially be developed as genetic markers of Alzheimer's disease. Long-tailed macaques exhibited the development of amyloid plaque in the aging brain based on the analysis of the gene expression profile of its biomarker. Furthermore, long-tailed macaques can be optimized for neurodegenerative models.
Standarisasi Penjaminan Kesejahteraan Hewan Untuk Kemajuan Riset Biomedis Fitriya Nur Annisa Dewi; Huda Shalahudin Darusman; Joko Pamungkas
Policy Brief Pertanian, Kelautan, dan Biosains Tropika Vol 4 No 4 (2022): Policy Brief Pertanian, Kelautan dan Biosains Tropika
Publisher : Direktorat Kajian Strategis dan Reputasi Akademik IPB University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.29244/agro-maritim.4.4.375-380

Abstract

Pengembangan berbagai obat, vaksin dan produk kesehatan memerlukan tahap penelitian dan pengujian pada hewan laboratorium/hewan coba. Kesejahteraan hewan selama pemeliharan dan penggunaannya dalam studi merupakan salah satu faktor penting yang menentukan kesahihan hasil, dan pemenuhannya semakin menjadi sorotan publik dan tuntutan global. Diperlukan aturan hukum, kebijakan dan pedoman nasional untuk menjamin implementasi prinsip kesejahteraan hewan yang terstandar di berbagai institusi di Indonesia, dan mengacu pada standar internasional. Kajian ini bertujuan menelaah aspek legal di Indonesia yang mengatur implementasi prinsip etik dan kesejahteraan hewan di bidang penelitian biomedis, dan mengidentifikasi kebijakan dan pedoman yang diperlukan untuk mencapai standarisasi penjaminan tersebut.
Development and Optimization of an Immunoassay for the Detection of Antibodies Against SARS-CoV-2 with In-house Recombinant RBD Protein Ratu, Safira Pinaka Pramestika; Mariya, Silmi; Noviana, Rachmitasari; Saepuloh, Uus; Darusman, Huda Salahudin
Makara Journal of Science Vol. 26, No. 3
Publisher : UI Scholars Hub

Show Abstract | Download Original | Original Source | Check in Google Scholar

Abstract

COVID-19 caused by SARS-CoV-2 poses a major threat to the global community, particularly in Indonesia. Countermeasures to prevent the spread of this disease have also been implemented, including the implementation of a vaccination program. An immunoassay technique that can be used to analyze antibodies that might develop following vaccination is the indirect enzyme-linked immunosorbent assay (ELISA). We produced the recombinant spike protein used in this study. The optimization comprised adjusted concentrations of spike recombinant protein (5 and 10 ng/mL), blocking agent (2.5% and 5%), and conjugate (1:1000 and 1:5000). The optimal conditions in this study included a spiked concentration of 10 ng/mL, a blocking agent concentration of 5%, sample dilution of 1:33, and a conjugate concentration of 1:1000. The intra-assay value of this optimized indirect ELISA was 7.3, and the inter-assay value was 5.3. The commercial MyBioSource kit and immunodiagnostic were utilized as a reference in the T-test, with P-values of 0 and 0.313, indicating that the recombinant protein in-house ELISA kit in this study demonstrated the same ability as the commercial immunodiagnostic kit in detecting SARS-CoV-2 antibodies, allowing it to be used for post-vaccination efficacy evaluation.
Kejadian nematodosis gastrointestinal pada monyet ekor panjang (Macaca fascicularis) di Pusat Studi Satwa Primata LPPM IPB Rosyid, Bahaudien; Darusman, Huda Shalahudin; Retnani, Elok Budi
Current Biomedicine Vol. 1 No. 1 (2023): January
Publisher : School of Veterinary Medicine and Biomedical Sciences, IPB University, Bogor, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.29244/currbiomed.1.1.33-45

Abstract

Background: The long-tailed macaque (Macaca fascicularis) is often used as a model animal in biomedical research. Nematodosis is common among non-human primates. Gastrointestinal nematodosis may affect study results or be a potential source of transmission to humans (zoonosis). Objective: The study aimed to identify the type, calculate the degree of infection, and the prevalence of gastrointestinal nematode worms in long-tailed macaques in captivity at the Primate Research Center, Institute for Research and Community Service (PSSP LPPM) IPB University. Methods: Faeces samples from 24 long-tailed macaques were examined using the McMaster and Kato-Katz simple flotation methods. Results: Nine (37.5%) of the 24 samples were infected by trichurid (20.8%), ascarid (12.5%), and strongylid (4.2%), each with an average degree of infection 833.3 ± 923.8, 32 ± 27.7, and 12 ± 17 eggs per gram of faeces (EPG), respectively. The prevalence of nematodosis in long-tailed macaques based on age group and sex was in infants 50%, children 50%, adolescents 16.7%, adults 33.3%, males 33.3%, and females 41.7%. Conclusion: The incidence and degree of gastrointestinal nematodosis infection in long-tailed macaques showed a moderate to low level. These results can be considered in maintenance management, biomedical research, and mitigation of potential transmission risk to humans.
Co-Authors A.S. Satyaningtijas Adi Winarto Adi Winarto Agik Suprayogi Ahmad Kamal Nasution Ahmad Subhan Ai Hertati, Ai Akhiruddin Maddu Andhika Yudha Prawira Andi Hiroyuki Angela Mariana Lusiastuti Apon Zaenal Mustopa Aprilyana, Lia Ardiansyah Nurdin Benvika Damiana Rita Ekastuti Dayana, Hepy Dewi Ratih Agungpriyono Dewi, Fitriya Nur Annisa Dondin Sajuthi DYAH PERWITASARI -FARAJALLAH Ekowati Handharyani Elok Budi Retnani Entang Iskandar Entang Iskandar Entang Iskandar Entang Iskandar Ernes Andhesfa Etih Sudarnika Fadla, Ilma Nadya Fauzi Abdurrahman Munggaran Femke Den Haas Ferdinandez, Mariana Firdaus, Moh Egy Rahman Fitri, Arni Diana Habib Ilham Maulana Al Adhim Hana Intishar Sawitri Hardo Basuki Harrold Halilintar MR Subu Taopan HERA MAHESHWARI Hidayaturrahmah Hidayaturrahmah I wayan Teguh Wibawan Ice Lusia Marta IETJE WIENTARSIH Ikramullah, Muh. Chaeril Indra Bachtiar Indra Bachtiar Indriawati, Iin Iqbal Ngabdusani Irawan, Shasmita Irma Herawati Suparto Jusak Nugraha Kanti Rahmi Fauziyah Karin katina Hasya Katrin Roosita koekoeh santoso Krisnayanti, Ni Putu Eka Lina Elfita Lina Elfita Linda Tjhin Lis Rosmanah Lis Rosmanah Loes (ML) Schure Maria Fatima Palupi Maria Fatima Palupi Maria Fatima Palupi Megawati Iskandar Mia Audina, Mia Muhammad Hamdan Ningrum, Emilna Mega Novi Mayasari Noviana, Rachmitasari Novriyandi Hanif Nugroho, Setyo Widi Pamungkas, Joko Perdhana, Ika Satya Permanawati Permanawati Puji Rianti Putri, Indah Aprianti Rachmitasari Noviana Rahmania Hanim Rahminiwati, Min Randall C. Kyes Rasyida Ulfa Ratna, Lita Tri Ratni Ernita Ratu, Safira Pinaka Pramestika Raymond R. Tjandrawinata Ridzki M.F. Binol Rifqiyah Nur Umami, Rifqiyah Nur Rika Sudranto Rizki Aulia Ansari Rosyid, Bahaudien Savitri Novelina Sela Septima Mariya Setyo Widi Nugroho Silmi Mariya Silmi Mariya SILMI MARIYA Silvy Thiyana Sony Hartono Wijaya Srihadi Agungpriyono Suduri, Adam Sukenda Sukenda Sumitro - Tambunan, Anggitha Tatang Mitra Setia Tatik Mufidah Tommy Langgeng Abimanyu Tri Wibawa Uus Saepuloh Uus Saepuloh UUS SAEPULOH Vinka Aftinata Kusumaputri Wartika Rosa Farida Wasmen Manalu WIDANARNI WIDANARNI Wira, Dwi Wahyuda Yuliana Yuliana Yuliana Zaelani, Bella Fatima Dora