HAYATI Journal of Biosciences

Antimigratory Activity of Brazilin-Containing Fraction from Caesalpinia sappan L. on MDAMB-231 Cells

Thu, 01 Oct 2020 00:00:00 +0700

Caesalpinia sappan is studied for several biological activities. The aim of this research is to determine the cytotoxic and antimigratory activities of Caesalpinia sappan active fraction in combination with cisplatin on human TNBC cells (MDA-MB-231). Caesalpinia sappan heartwood was extracted with methanol. Then, several fractions of the methanol extract were obtained by using a liquid-liquid extraction method followed by column chromatography. The cytotoxicity was determined using MTT assay. Synergistic effects were analyzed by calculating the combination index (CI). Migration was examined using wound-healing assay. Levels of MMP2 activity were determined with gelatin zymography assay. The results showed that most of the fractions included in this study exhibited cytotoxic effects against MDA-MB-231 cells, and C fraction demonstrated the highest cytotoxic activity of all fractions. The combination of C-cisplatin revealed a synergistic inhibitory effect on MDA-MB-231 cell growth (CI<1). Furthermore, C fraction, alone and in combination with cisplatin, inhibited migration of MDA-MB-231 and suppressed MMP2 activity. The C fraction isolated from Caesalpinia sappan increased the cytotoxic and antimigratory activities of cisplatin on MDA-MB-231 cells. Based on these findings, the potential of Caesalpinia sappan to act as a supportive agent in metastatic TNBC treatment with cisplatin warrants further exploration.

Biological Conservation of Molluscs Based on Spatial and Temporal Distribution in Tropical Tidal Lake, Medan-Indonesia

Thu, 01 Oct 2020 00:00:00 +0700

The tidal lake ecosystem is formed by the flow of the Belmera River and seawater through a tidal process along a course of 12 km from Belawan (Malacca Strait). This lake has a tidal lake that has the characteristics of fresh water to brackish water and Mollusc communities from freshwater species to brackish water species. This study was performed to develop a mollusc conservation strategy based on the spatial and temporal distribution of Mollusca in the tides. The study was conducted from September 2018-August 2019. Mollusc samples were taken every month at high tide and low tide using a Petersen grab tool. Nine Mollusca species, consisting of three bivalves and six gastropods, were identified. Polymedosa expansa is a mollusc species that is considered for conservation. The population size of P. expansa is small, and its distribution is limited to the northern lake. P. expansa was found only in September at high tide and in December at low tide. Competition among Mollusca and habitat availability are obstacles to the survival of P. expansa. The habitat of P. expansa is mangrove, and therefore a conservation approach was carried out through improvement of mangrove quality.

Integration Stability of sHBsAg-Multi Expression Cassettes in Pichia pastoris GS115 during Methanol Induction

Thu, 01 Oct 2020 00:00:00 +0700

Hepatitis B is the major health problem worldwide including in Indonesia. Vaccination is the best prevention strategy for the disease. For the purpose of vaccine development and to decrease drug import, production of Hepatitis B Virus (HBV) small surface antigen (sHBsAg) from Indonesian HBV subtype is needed. The recombinant protein production can be conducted by integrating multi expression cassettes of sHBsAg gene in Pichia pastoris chromosome using gene replacement method. Such integration method turns out to allow loss of foreign gene from chromosome by excisional recombination-mediated looping out. This research was aimed to determine integration stability of four copies of sHBsAg expression cassette in P. pastoris GS115 chromosome inducted with 2% methanol in FM22 medium. The methanol induction was conducted twice at 63-h and 75-h. Integration stability determination was conducted qualitatively using PCR and quantitatively using qPCR absolute quantification. A band of 208 bp with similar intensity was observed after amplification of genomic DNA. All samples generated the same Ct value of around 22 with four copies of sHBsAg gene per genome. The result from this experiment shows that integration of four copies of sHBsAg expression cassette in P. pastoris GS115 chromosome is stable during methanol induction.

Cysteamine in Maturation Medium Enhances Nuclear Maturation and Fertilization Rate of Sheep Oocytes In Vitro

Feni Dwi Kartika Gulo, Ni Wayan Kurniani Karja, Mohamad Agus Setiadi — Thu, 01 Oct 2020 00:00:00 +0700

Low nuclear maturation and fertilization rate is one obstacle in the in vitro embryo production which decrease embryo yield. This problem is presumable related with high production of reactive oxygen species (ROS) during maturation process. Glutathione (GSH) as an antioxidant is well known to overcome effect of ROS production. GSH synthesis in the cytosol part of the oocyte cytoplasm is influenced by cysteine availability. It is therefore, this research was conducted to evaluate the ability of cysteamine to provide cysteine availability as GSH precursor on the nuclear maturation and fertilization rate of sheep oocytes. Results of this experiment revealed additional cysteamine at 150 µm and 200 µm could significantly improve nuclear maturation rate. On the other side, although additional of cysteamine at 50 µm could not improve nuclear maturation rate, however 50 µm cysteamine in the maturation medium could significantly improve the fertilization rate. Based on those experiment results, it seems that the additional cysteamine might be improve not only GSH availability but also the oocyte quality which characterized the ability of pronuclear formation. This finding strongly suggested that additional cysteamine in the maturation medium could improve nuclear maturation and fertilization rate of sheep oocytes.

An Optimization Model for Environmental Ergonomics Assessment in Bioproduction of Food SMEs

Mirwan Ushada, Hani Febri Mustika, Aina Musdholifah, Tsuyoshi Okayama — Thu, 01 Oct 2020 00:00:00 +0700

Environmental ergonomics in bioproduction of food Small Medium-sized Enterprises (SMEs) become a concern and need to be optimized. An optimization model was developed using a Genetic Algorithm (GA). The weight of an Artificial Neural Network Model was used as a fitness function for GA. The research objectives were: 1) To design an environmental ergonomic assessment system for bioproduction of Food SMEs, 2) To develop an optimization model for environmental ergonomic assessment using a Genetic Algorithm. GA is utilized to search optimal set points of environmental ergonomics based on the predicted fitness values. Each chromosome of GA represents the environmental ergonomics value. The parameters were heart rate, bioproduction temperature, distribution of bioproduction relative humidity and light intensity. The target of the optimization model was the bioproduction temperature set points. The research result indicated the model generated optimum values of environmental ergonomics parameter in bioproduction of food SMEs. The parameters could be used to provide standard workplace environment for the sustainability of food SMEs.

Protective Signatures of Roselle (Hibiscus sabdariffa L.) Calyx Fractions against Staphylococcus aureus in Drosophila Infection Model

Thu, 01 Oct 2020 00:00:00 +0700

The rise of antibiotic-resistant Staphylococcus aureus-related clinical cases is an alarming chronicle for global communities. This research was conducted to examine the antistaphylococcal effect of roselle (Hibiscus sabdariffa L.) calyx fractions in the Drosophila model. In the infection experiment, wild-type and immunodeficient Drosophila were pricked with S. aureus and subsequently subjected to fly survivorship and colony-forming assays, in the presence or absence of roselle calyx fractions. The Involvement of immune stimulation in the host antibacterial protection was assessed in vitro using cell-based luciferase reporter assay and in vivo using RT-qPCR analysis on adult flies. A declining rate of fly survivorship and augmentation of bacterial growth were observable in S. aureus-infected wild-type flies but subject to improvement in the presence of roselle calyx fractions. Cell-based analysis revealed the absence of host immune stimulation via Drosophila Toll pathway and roselle calyx fractions-treated immune-deficient flies lacking for components in the Toll pathway were protected from infection-induced early death phenotype and harbored reduced number of S. aureus colonies. Overall, our data confirmed the in vivo anti-staphylococcal activity of roselle calyx fractions in Drosophila infection model and such protective signature was devoid of host immune stimulation.

Implementation of SExI–FS (Spatially Explicit Individual-based Forest Simulator) Model using UAV Aerial Photo Data Case Study: Jatinangor ITB Campus

Thu, 01 Oct 2020 00:00:00 +0700

Landscape architecture affected by interaction between built and natural environment such as vegetation. Nowadays, landscape architects are using 3D city models for simulations, which requires highly dynamic and time-varying attributes. 3D city modelling structure has been standardized by CityGML, although researches that are related to the storing of dynamic data had been conducted for the past years, it has not been supported by any standard until this very moment. In dynamizer, it is added as a data structure into a CityGML structure that is already existed, although the existing structure is a static one. Kolbe’s research on dynamic data using CityGML called dynamizer could use the spatial data in more dynamic way by changing its geometric, thematic, or appearance data, but its purpose is not specific for trees or vegetation. In this paper, a method of simulating the vegetation growth using SeXI-FS will be discussed to show the dynamic changes that happen in vegetation as part of the dynamic changes in landscape architecture. The result of this research will be used to address the importance of information on vegetation by studying its changes in Jatinangor ITB Campus and as initial research to build dynamizer in CityGML for landscape architecture.

Quality Improvement of Semi-Wet Terasi by Optimizing the Starter Culture Ratio of Controlled Fermentation

Arisa Sato, Dea Indriani Astuti, Sastia Prama Putri, Eiichiro Fukusaki — Thu, 01 Oct 2020 00:00:00 +0700

Terasi is a traditional fermented shrimp paste used in Indonesian dish as condiments. Due to its affordability, the paste is widely consumed among the general population, and thus has a great impact in Indonesia. Currently, small-scale or home industry is common for terasi production, and natural fermentation process is the traditional method. Fermentation process is considered complete when desired aromatic odors are obtained. However, this makes the fermentation process subjective, because the decision is solely dependent on the producer. Additionally, natural fermentation poses a higher risk for contamination of microbial pathogens. As a result, the quality of the final product varies greatly from region to region. Therefore, it is necessary to improve the quality of terasi by means of controlled fermentation. Hence, the objective of the research is to optimize the controlled fermentation condition of terasi by determining the most optimal ratio of mixed starter culture. Optimal fermentation conditions were determined by analyzing the effect of the various starter inoculum on the inner microbial community, and results indicated that mixed culture of Staphylococcus saprophyticus, Bacillus subtilis, and Lactobacillus murinus with ratio of 2:1:2 was the most effective for suppressing the growth of unwanted microorganisms. The difference in the microbial composition also resulted to a change in the metabolite profile of terasi.

The Genetic Structure of Nilaparvata lugens (Stal.) in Java Populations

Ruth Martha Winnie, Rika Raffiudin, I Nyoman Widiarta, Aunu Rauf — Thu, 01 Oct 2020 00:00:00 +0700

The brown planthopper Nilaparvata lugens (Stal.) causes hopperburn in the rice field. The wing macropterous adults of this insect have capability for long distance flight. We aimed to analyze the genetic structure of six N. lugens populations in Java inferred from the combined COI-COII mitochondrial and ten loci of microsatellites markers. This study found low levels of nucleotides and high levels of haplotype diversity of the planthopper. The demographic test might indicate the genetic bottleneck history of N. lugens population in Java. The genetic diversity analyzed by using microsatellite markers also showed high levels of heterozygosity (Ho>He) that indicated an isolated-breaking effect in the six populations in Java. Moreover, we found a homogenous genetic structure of N. lugens based on pairwise fixation indices (Fst) analyzes that appears to be maintained by high levels of gene flow and showed no correlation between genetic and geographical distance. Importantly, these studies also support accurate information of widely distributed and genetically intermixed among N. lugens across Asia populations. Thus, our results support the theory of long-distance migration among N. lugenspopulations. The genetic structure information of N. lugens in Java could support regional management, such as the new controlling strategies based on forecasting systems.

Differentiation of Cardiomyocytes and Identification of Cardiac Conduction System Connexins Derived from Bone Marrow Mesenchymal Stem Cells of Macaca nemestrina

Thu, 01 Oct 2020 00:00:00 +0700

Bone marrow mesenchymal stem cells have been widely used, because plasticity, specific surface markers, self-renewal to transform into various lineages including cardiomyocytes. Information about the connexin (Cx) cardiac conduction systems of the pigtail macaque (Macaca nemestrina) is limited. This study aimed to evaluate cardiomyocyte differentiation from bone marrow mesenchymal stem cells of pigtail macaques and to clarify the Cx cardiac conduction system. Bone marrow aspirates were obtained from the proximal humerus of four adult male pigtail macaques, collected into heparinized tubes, then centrifuged to obtain mononuclear cells that were isolated and cultured in an incubator. After these cells reached 70–80% monolayer confluency as homogeneous fibroblast-like cells, they were subcultured. On the second subculture passage, the cells were pelleted to extract the mRNA, which was analysed by reverse transcription–polymerase chain reaction, and then cultured for a third passage. Cells were positive for CD73 and CD105 and the reference gene glyceraldehyde-3-phosphate dehydrogenase, and negative for CD34 and CD45. Osteogenic, chondrogenic, adipogenic, and cardiomyocyte differentiation was confirmed based on specific staining. The pigtail macaque bone marrow mesenchymal stem cells can be isolated and subcultured. The transcription of genes and translation of proteins of the connexin cardiac conduction systems was successfully identified.