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Sea cucumbers or holothurians are marine invertebrates, belonging to the phylum Echinodermata (kingdom Animalia). In Asia, they are commonly used as food, while they are applied in traditional medicine as well. A wide range of secondary metabolites from these animals has been reported, but triterpene glycosides are the most predominant and most studied and are considered as valuable chemotaxonomic markers. With respect to this, the genus Holothuria is an important genus of sea cucumbers, given the high number and variety of triterpene glycosides previously reported. Holothuria triterpene glycosides are typically composed of a 3β-hydroxyholost-9(11)-ene aglycon and a glycosidic moiety comprised of one up to six sugar units, connected to position 3 of the sapogenin. In the past decade (2010–2020) as much as 63 saponins were identified for the first time in the Holothuria genus, of which 36 had not been reported before. Identification was mainly carried out by LC–MS (liquid chromatography—mass spectrometry) analysis, while also NMR (nuclear magnetic resonance) spectroscopy is often applied for structure elucidation. However, while carrying out a literature search on these compounds, various inconsistencies were observed. Therefore, this review intends to provide an overview of typical signals that can occur in NMR and MS data, accompanied with an extensive explanation of their interpretation, in order to facilitate any future research on identification/structure elucidation of Holothuria saponins. Moreover, the data published in the past decade were critically reviewed and various questionable results are discussed.

Salmonellosis that attacks poultry, caused by Salmonella enteritidis is a cause of food borne disease in zoonotic humans. Treatment of this disease uses antibiotics but if used irrationally it can result in the emergence of multi-drug resistant bacteria. At present many treatments use herbs which are considered to reduce the negative effects of antibiotics and are environmentally friendly, one of which is coffee. Green coffee is considered to have many active ingredients such as chlorogenic acid (CGA), alkaloids, tannin, polyphenols and polysaccharides which are useful as antimicrobial, anti-inflammatory and enhances the body's immune system. The study used 60 heads ISA Brown day-old chicks (layer strain) chickens artificially infected with S. enteritidis bacteria with concentration of 10 8 CFU/ml, consisting of negative controls (healthy chickens, without coffee, positive control (chickens were given), T1, T2 and T3 was given different doses of coffee extract with 500 mg/kg BW, 1000 mg/kg BW and 1500 mg kg BW, respectively. The treatments with coffee extracts were given on days 3-15, then infected with 0.5 ml bacteria on day 16 and 60 chickens were necropsied at day 18 for histopathology. Data on relative levels of CD8 Tc cells were calculated using the flowcytometer test and analyzed quantitatively using the One Way ANOVA with a confidence level of 95% while the jejunum histopathology was observed using a microscopy and analyzed descriptively. Results showed that Robusta coffee extract from Lampung can act as an antibacterial by increasing CD8 + T cells and repairing jejunum histopathology

Mycobacterium tuberculosis (M. tuberculosis) is the causative agent of tuberculosis in human. One of the major M. tuberculosis virulence factors is early secretory antigenic target of 6-kDa (ESAT-6), and EccB5 protein encoded by eccB5 is one of its components. EccB5 protein is a transmembrane protein in ESX-5 system. The aim of this study is to explore the characteristics of wild-type EccB5 and its mutant form N426I. We expressed the EccB5 protein by cloning the mutant and wild-type eccB5 gene in Escherichia coli (E. coli). We compared the protein structure of wild type and mutant form of EccB5 and found changes in structure around Asn426 (loop structure) in wild type and around Ile426 (β-strand) in the mutant. The truncated recombinant protein of EccB5 was successfully cloned and expressed using plasmid pCold I in E. coli DH5α and E. coli strain Rosetta-gami B (DE3) and purified as a 38.6 kDa protein by using the affinity column. There was no detectable adenosine triphosphatase activity in truncated forms of EccB5 and its mutant. In conclusion, our study reveals successful cloning and protein expression of truncated form of eccB5 gene of M. tuberculosis. EccB5 protein in ESX-5 system may be an important membrane component involved in the transport machinery of type VII secretion system, which is essential for growth and virulence.

Background and Aim: Gliricidia sepium is a medium-sized leguminous plant found widely in tropical to subtropical areas. It has been used as a medicinal ingredient and in rodenticides by local communities in both Indonesia and the Philippines. This study aimed to investigate the wound healing effects of an ointment containing G. sepium leaves on inflammatory cells using a rat model. We also determined its effect on the expression of interleukin (IL) 6 and IL-1β. Materials and Methods: We used 16 Wistar male rats aged approximately 2 months and weighing 150-200 g. They were divided into four treatment groups (T1, positive control; T2, negative control; T3, wounds treated with G. sepium from Indonesia; and T4, wounds treated with G. sepium from the Philippines), and the ointment therapies were applied to wounds for 3 days. Hematoxylin and eosin staining was performed to examine the inflammatory cells microscopically. IL-1β and IL-6 expression were observed immunohistochemically. Results: G. sepium leaves significantly (p

Background and Aim: Anisakis is a global challenge for a fish product which may lead to a decrease in economic value and consumers' preference. Skipjack (Katsuwonus pelamis) in Kupang, Nusa Tenggara Timur, Indonesia, have important economic value for local fisheries. Anisakis typica is one of the Anisakis species which potent to induce an allergic reaction. However, the study about A. typica involved in the dendritic cells (DCs), T helper 1 (Th1), T helper 2 (Th2), and regulatory T cells (Tregs) is still limited. This study aimed to analyze the dynamic changed of the immune system including DCs, CD4+ T cells, and Tregs after 1 week of A. typica sensitization. Materials and Methods: Twenty-four male Balb/C mice were randomly divided into four groups (n=6), mice treated with crude A. typica extract (CAE) 50, 75, and 100 mg/kg BW, respectively. CAE was given orally per day for a week. At the end of the experiment, the animals were sacrificed and the spleen was collected. DCs were labeled as CD11c+ interleukin-6+ (IL-6+); CD4+ T cells were distinguished as Th1 (CD4+ interferon-γ+ [IFN-γ+]) and Th2 (CD4+ IL-4+ and CD4+ IL-5+); Tregs were labeled as CD4+CD25+CD62L+. The expression of each cell was determined by flow cytometry. Results: Our result described that CAE elicits CD11c+ IL-6+, CD4+ IFN-γ+, CD4+ IL-4+, and CD4+ IL-5+ and reduces CD4+CD25+CD62L+ significantly (p

The aim of this research was to study the antidiabetic activity of root extract of Ruellia tuberosa L. on rats ( Rattus novergicus ) induced by multiple low doses of streptozotocin. The parameters investigated were blood glucose levels, protease activities, and the histopathologic profiles of the kidneys. Three groups of rats, i.e. control, diabetic, and treatment, were used. R. tuberosa L. extracts were given orally for treatment at a dose of 250 mg/kg body weight per day for 21 consecutive days. The induced diabetes group treated with root extract of R. tuberosa L. (group III) had 60.3% lower glucose levels (119 mg/dL) compared to the group with induced diabetes only. Moreover, antidiabetic activity of R. tuberosa L extract was evidenced by the 55% lower protease activity in the treatment group compared to the untreated diabetic group. Finally, a positive effect from root extract of R. tuberosa L. was shown from the less damaged renal glomerulus and smaller gap between extracellular matrices and epithelium cells in the histopathologic profiles of the kidneys in the extract treatment group. These results show the potential of root extract of R. tuberosa L. as a natural product anti-diabetic agent.

Lead (Pb) is one of the heavy m metals polluting the environment which is harmful to t the organism. Pb accumulation triggers free radic cals and disrupts the body's metabolism. Pluche indic ca leaf extract is reported contain flavonoids whiich act as antioxidants and can counteract free radiccal attacks. The purpose of this study was to de etermine the role of Pluchea indica leaf extract preve entive therapy in counteracting free radicals due too exposure to Pb in rats (Rattus norvegicus). Wistar mmale rats aged of 8-10 weeks with a body weight of about 200-250 grams. Rats were divided into 5 tr reatment groups, were group A as negative contr rol, group B as positive control induced by lead (Pb)) 10 mg/rat/day, groups C,D,E given with ethano olic extract of Pluche indica leavesof 200 mg/kgBW, , 400 mg/kgBW, and 600 mg/kgBW and lead (Pb b) exposure of 10mg/ rat/day. Kidney, liver and jejunaal histopathology using HE staining was observed microscopically. The results showed that the administtration of Pluche indica leaves extract could preve ent tissue damage to the kidney, liver and jejunum in rats exposured to Pb. Dose of 600 mg/kgBW show wed histopathological improvement in kidney, liver annd jejunum. The greater the dose, the better prev ention of tissue damage as indicated by tissue condition resembling those of control rats. The conclusion of this study, the Pluchea indica leaf extract has the potential to prevent organ damaged exposed to lead.

Chronic exposure to UVB irradiation can caused premature skin aging (photoaging), as it can trigger the production of ROS and initiate both oxidative stress in cell membrane and affect the molecular pathway. This paper figured out the potential of oral and topical administration of C. asiatica extract on hairless rats skin tissue exposed by UVB based on expression of TGF-β1, MMP-1 and MDA dermis. A total of 30 male-hairless rats (Rattus novergicus) divided into 6 experimental groups, exposed by UVB with total dose 840 mJ/cm2 for four weeks. Topical gel therapy administered twice a day, 20 minutes before and 4 hours after irradiation. Oral therapy administered once a day before irradiation. Expression of TGF-β1, MMP-1 and MDA dermis observed by immunohistochemistry. The result showed that oral and topical therapy of C. asiatica extract was capable of inhibiting the increase of MMP-1 and MDA expression and the decrease of TGF- β1 significantly (p

Background: Hypertension is a disease that has the potential to become the most significant cause of death in the world in 2020. Hypertension caused by an imbalance of the renin angiotensinogen system. Viscera hydrolyzate extract has an anti-hypertension activity. The research aimed to determine tilapia viscera hydrolyzate extract’s ability to reduce blood pressure, suppressing the expression of TNF-α and IL-6. Viscera hydrolyzed with alcalase and extracted to obtain viscera bioactive crude extract (VisBC).Methods: Spontaneously hypertensive rats induced by DOCA-salt, gave VisBC therapy. 15 Wistar rats divided into 5 treatment groups, namely normal control (without induced DOCA-salt), positive control (SHR-Spontaneous Hypertensive Rat = induced by DOCA-salt), comparative control (SHR + Captopril 5 mg/kg), VisBC 1 (SHR + BC 150 mg/kg) and VisBC 2 (SHR + BC 300 mg/kg), where each treatment consisted of 3 replications. Each treatment measured for blood pressure, TNF-α and IL-6 expression. The data represented as mean ± SD and analyzed by ANOVA (p less than 0.05) in any difference, a subsequent Duncan test. Data analyzed by SPSS 20.0 for Windows 10.Result: The results showed the treatment of VisBC could reduce systole (19.64%) and diastole (29.15%) blood pressure. Also, VisBC inhibited the expression of TNF-α (8.08%) and IL-6 (8.39%). The VisBC of tilapia with a dose of 150 mg/kg could inhibit the expression of TNF-α and IL-6, thereby affected the decrease in blood pressure (systole and diastole) in rats induced by DOCA-salt. This research proved that fish waste could produce bioactive as anti-hypertension.

Human Papilloma Virus (HPV) infection suggested could induced gene hypermethylation. Hypermethylation occurs on the p16 tumor suppressor gene promoter resulted in silencing tumor suppressor gene so that the gene is inactive and contributes to the malignancy of cancer cells. Tea parasites or Loranthaceae are known to be a plant that has an anticancer potency, correlated to flavonoids content. Flavonoid from Scurrula atropurpurea (Blume) Danser (SAD) suggested have a role in inhibiting methylation towards p16. This research objective was to identify the potency of Lawang - East Java SAD flavonoids compound towards Hela cells. In this study, we examine flavonoids from SAD that been extracted in n-hexane, chloroform, ethanol solvent, using TLC Chromatogram, and LCMS. We also examined the cells proliferation using MTT assay, the apoptotic level was examined by Tunnel, and then completed with methylation level detection using bisulfite direct sequencing method. The result showed that flavonoids that found were: flavanone, dihydro flavonol, and flavon, dihydro flavonol, flavanone and catechin, and flavonol, flavon, and EGCG, respectively n-hexane, chloroform and ethanol fraction. Flavonoids have proliferation inhibitor and apoptotic inductor potency via inhibition of p16 promoter of HeLa cells methylation. SAD flavonoids in the n-hexane and chloroform fraction shown the most potent in methylation inhibition.

Widyaningrum T, Suharjono S, Ardyati T, Aulanni’am A. 2020. Diversity and potency of indigenous yeast from some palm juices for bioethanol production. Biodiversitas 21: 318-325. Main energy source commonly used by the community comes from fossil energy, especially petroleum. The crisis of energy showed that Indonesia's fossil energy reserves are limited. Based on this fact it is important to develop alternative energy that environmentally friendly and sustain, especially bioethanol. The objective of this research was to diversity and potency of indigenous yeast from palm juice of Arenga pinnata Merr., Cocos nucifera L., Nypa fruticans Wurmb., and Borassus flabellifer L. for bioethanol production. Yeast from juice of the four species of palm was isolated using Yeast Malt Extract Agar (YMEA). Those isolates were screen base on ethanol production in coconut water media. The potential of yeast isolates was identified base on 18S DNA sequence similarity. The four potential isolates of yeast were A3A from A. pinnata, K1A from C. nucifera, N3E from N. fruticans, and S1A from B. flabellifer have highest production of ethanol are 8.6 %, 14.2 %, 9.0 %, and 9.2 % respectively. Four potential isolates of yeast were A3A and N3E, K1A, and S1A have relationship with Saccharomyces cerevisiae NRRL Y-12632T, Pichia manshurica IFO 10726T, and Candida tropicalis ATCC 20615 respectively.

Background: Mycobacterium tuberculosis Complex (MTBC) is a group of Mycobacterium that causes tuberculosis (TB). TB is an infectious disease that remains a global health problem. Indonesia is one of the five countries in the world where TB is the most prevalent and became the country with tle second largest rate of TB in 2014 and 2015. MTBC has high pathogenicity that can cause infections in animals and humans. The most common route of transmission is via airborne droplet nuclei and contact with animals or humans infected with TB. MTBC has many virulence factors. One of these factors is EccB5 that is encoded by eccB5 gene. EccB5 is a transmembrane proteinconserved membrane protein and could play a role in inducing damage in host cells, macrophage infection, and may correlate with active disease. The characterization of eccB5 gene needs to be studied to determine the nucleotide sequences, which may be associated with active disease. The aim of this research was to analyze the nuclotide sequences of eccB5 gene of MTBC from suspected pulmonary tuberculosis patients, SNPs of eccB5 gene and possible correlation with the disease, especially in Indonesia. Materials and Methods: Samples were collected from the Tuberculosis Laboratory, Clinical Microbiology of Dr. Soetomo Hospital Surabaya Indonesia. DNA extraction used boiling extraction method and continued nucleic acid amplification using PCR techniques. Primer pairs used eccB5 SK.. The positivity of DNA specific revealed amplicon in 1592 bp. PCR product was sequenced by 1st Base (First BASE Laboratories Sdn Bhd, Selangor, Malaysia). The sequence analysis used Genetyx-Win version 10.0 (Genetyx Corporation, Tokyo, Japan). Results: Total isolates of Mycobacterium spp. were 28 and those that showed positive MTBC were 24 isolates and 4 nontuberculosis mycobacteria (NTM) using immunochromatographic test (ICT). The amount of homology from MTBC using blast NCBI was 99%-100%. Two SNPs were found in position c.1277 which revealed replacement of amino acid in 426 of codon position. Conclusion: The sequence of eccB5 gene of MTBC showed high significant homology, while proposed nonsynoymous single nucleotide polymorphisms (nsSNP) may associated with clinical outcomes. Keywords: Pulmonary Tuberculosis, Mycobacterium tuberculosis Complex, eccB5 gene, SNPs

Diabetes mellitus type 1 (DMT1) is an insulin deficiency disorder caused by pancreatic damage characterized by chronic hyperglycemia. Animal modeling of DMT1 using inducting Multiple Low Dose Streptozotocin (MLD-STZ) elevat levels of free radicals and inflammation in pancreatic organs which then damage β cells. Ruellia tuberosa L. is a plant containing triterpenoids from the class of antioxidant and anti-inflammatory flavonoids. This study used three groups of rat: negative control group, positive control and 240 mg/kg bw therapy group. Study are based on changes MDA levels, blood glucose and histologic of pancreas. MDA level determining with TBA method, glucose levels using EasyTouch GCU and histopalogy of pancreas using Hematoxylin-Eosin staining. Statistical analysis of MDA and blood glucose (α = 0.05) showed significant decreases in MDA and blood glucose levels. Based on the results of histologic observations of pancreas descriptively showed a decreas in cell damage on Langerhans island. Thus, the provision of root extract of Ruellia tuberosa L. can inhibit the damage in rat DMT1 model so that it can be used as therapy of diabetes mellitus.

This research studies the effect of anthocyanin from purple sweet potato in the cerebellum of stroke ischemic rats by MCAO (Middle Cerebral Artery Occlusion) technique. This technique was carried out by ligating the blood flow in ECA and CCA region for 3 hours, followed by reperfusion. The MCAO technique proved as a technique for preparing ischemic stroke rats. This technique induces releasing of BDNF and VEGF in the cerebellum of ischemic stroke rats. The level of BDNF measured by ELISA technique and VEGF expression used immunohistochemistry technique. The results showed that anthocyanin from purple sweet potato increased the level of both BDNF and VEGF expression in the cerebellum in ischemic stroke rats. It is suggested that anthocyanin could be used as a therapeutic agent for ischemic stroke.

The larvae of Anisakis were living parasites and made marine mammals, birds and reptiles the definitive host. Identification of Anisakis larvae using morphological observation methods will be difficult, especially if there are only a few samples that can be identified. PCR is an identification method using DNA from a small sample quantity and can provide DNA sequence samples. This study aimed to determine the type and infection level of Anisakis sp. at skipjack tuna (Katsuwonus pelamis) from the Oeba Fish Auction (TPI) Kupang City morphologically and molecularly. Morphological analysis results of 30 Anisakis larvae showed the body parts of Anisakis larvae, namely the head, digestive tract, and tail. The infection of Anisakis nematodes in skipjack tuna found five individual nematodes in muscle tissue, 59 individuals in stomach tissue, and 1991 individuals in internal organs. Alignment results between isolates At1 and At2 against isolates A. typica comparing (outgroup), isolate At1 and At2 have high homologs. Based on the results of the study concluded that the type I Anisakis isolated from skipjack tuna (Savu Sea) was Anisakis typica.

The production of Tilapia increased the impact on the increase of fish waste, such as viscera. One of the efforts to overcome and reduce fishery waste are by using them optimally. One of the efforts is hydrolysis technology, which can produce extracts that have bioactivity. This research aimed to find out and obtain an overview of the chemical components in the protein hydrolyzate extract from viscera of Tilapia (Oreochromis niloticus). The screening was performed for flavonoids, alkaloids (Meyer, Dragendrof, Bouchardat), tannins, terpenoids (steroids, triterpenoids), polyphenols, and saponins. The color intensity or the precipitate formation was used as analytical responses to these assays. The results of screening and identification showed the presence of chemical components of alkaloids, tannins, triterpenoids, polyphenols, saponins from the protein hydrolyzate extract of viscera of Tilapia (Oreochromis niloticus), on the other hand, flavonoids and steroids were not detected. These chemical components have the potential bioactivity as antioxidants, anti-nociceptive, anti-inflammatory, anti-HIV, anti-fungal, cardioprotective, immunoregulative, anti-malarial, anti-inflammatory, Cerebro-protective, anti-mutagenic, vaso-relaxing, anti-bacterial, anti-anxiolytic, analgesic, anti-nociceptive and anxiolytic. The extract has the potential to be developed in the field of health as pharmaceutical or nutraceutical products.

Probiotic of Lactobacillus casei Shirota strain is lactic acid bacteria that have benefits to enhance immunity system, as an antioxidant and has the ability to decrease the cholesterol level. The aim of this study was to determine the potential of Lactobacillus casei Shirota strain probiotic toward total cholesterol decreasing and SOD activity increasing in rats’ aorta with a high-cholesterol diet. This study used 25 male rats of Rattus novergicus Wistar strain that will be divided into 5 groups. The 1 st group as the negative control, and the 2 nd group as positive control. The positive control group was induced by high-cholesterol diet contained quail egg yolk, pure cholesterol, cholic acid and waste oil that were given for 3 weeks. The 3 rd , 4 th and 5 th group were treated by probiotic Lactobacillus casei Shirota strain with variation dose of 3.6 x 10 7 , 7.3 x 10 7 and 10.9 x 10 7 cells/kg bodyweight/day. The results showed that probiotic of Lactobacillus casei Shirota strain therapy dose of 10.9 x 10 7 cells/kg bodyweight/day can decrease the total blood cholesterol of rats’ serum that were given with high-cholesterol diet of about 32.0%. In addition, the probiotic of Lactobacillus casei Shirota strain therapy also increase the activity of SOD about 50.70%.

UVB generates the production of Reactive Oxygen Species (ROS) and decrease antioxidants enzymatic excessively. Both of these biological effects caused photo-aging. Excessive ROS production afforded overexpression of AP-1 as a major regulator of photoaging. This paper figured out the potential of Aloe vera extract as topical gel treatment on the UVB-induced skin photo-aging in twenty male wistar hairless rats (Rattus novergicus) divided into 2 groups. First group was induced by UVB and the second was induced by UVB and topical gel extract Aloe vera treatment. Each group was given treatment for 4 weeks. The expression of MDA and SOD were measured with immunohistochemistry. The result showed that the topical gel therapy decreased the MDA expression and increased SOD expression significantly (p < 0.01). The conclusion from this study was Aloe vera extract had potentially as an alternative topical treatment of photoaging.

This study aimed to determine the fermented milk of Lactobacillus casei Shirota strain potential on cholesterol levels, SOD activity, LPL enzyme activity, and cardiac histopathology. This study used 25 rats (Rattus novergicus) Wistar strain aged 2-3 months, weighing 150-200 grams. These rats were divided into 5 groups: negative control group, positive control group with high-cholesterol diet for 21 days, and therapy group with 1.0 ; 1.5 ; 2.0 mL/rat/day of fermented milk for 14 days. The analysis methods used in this study are One Way ANOVA and Tukey. The results showed that the fermented milk of Lactobacillus casei Shirota strain had a strong effect on blood cholesterol levels and increasing the SOD activity. This study also showed that the fermented milk of Lactobacillus casei with 2.0 mL/rat/day is the most effective dose to decrease the total cholesterol and increase the SOD activity.

Shigella flexneri is the most common causal agent of shigellosis. Its pili are composed of pili protein subunits. Adhesion molecules can be found on the pili and outer membrane proteins (Omp). A hemagglutination reaction can be used for screening of adhesion molecules. Objectives: The purpose of this study was to determine the molecular weight of the pili protein subunits and outer membrane proteins of S. flexneri that act as hemagglutinin proteins, and to prove whether there is a cross-reaction between antibodies against hemagglutinin pili protein subunits and outer membrane proteins of S. flexneri. Methods: Pili protein subunits were isolated using pili bacteria cutters, and the outer membrane proteins were solubilized and obtained using sodium dodecyl sulfate 0.05% as detergent for Omp isolation. The hemagglutination reaction used mice erythrocytes. The cross reactions between subunit pili proteins were conducted by Western blot and Dot blot. Results:. Antibodies against hemagglutinin sub unit pili protein 18 kDa responded to pili protein subunits 18 kDa; 23 kDa; 34 kDa; and 53 kDa; and Omp 23 kDa and 27 kDa. Omp and subunit pili proteins S. flexneri consists of several identical epitopes that were responsible for the similarity of the response profile in the cross-reactions of antibodies.

Cellulase enzyme is a multi-enzyme system consists of endo-β-1,4-glucanase (CMCase; EG; EC 3.2.1.4), exo- β-1,4-glucanase (cellobiohydrolase; CBH; EC 3.2.1.91), and β-1,4-glucosidase (GH; EC 3.2.1.21). The enzyme system works synergistically to degrade cellulosic substrates to glucose. The endo-β-1,4-glucanase enzyme has an important role in the initiation of cellulose hydrolysis. The exploration of endoglucanase is still continuing in order to achieve the best activity and efficiency. The aim of this study was to characterize the endoglucanase-encoding gene of indigenous Bacillus subtilis, a cellulolytic bacterium isolated from rice bran, by in silico analysis. The endoglucanase gene was isolated using forward (5’-TTCGCAGCAAGTGTAACGGA-3′) and reverse (5′-CTCGCCGTACATCGCATAGT-3′) primer, and amplified through PCR in this following condition; pre-denaturation at 94°C for 4 min, denaturation at 94°C for 30 sec, annealing at 54°C for 30 sec and extension at 72 °C for 1 min, the final extension was performed at 72°C for 2 min, all repeated to 35 cycles. The endoglucanase gene of indigenous Bacillus subtilis from rice bran comprises of 862 bp (246 amino acid). In silico analysis showed that endoglucanase gene of Bacillus subtilis has a catalytic cellulase domain constituted of BglC gene and CBM (Cellulose Binding Module). The binding region of the enzyme is located at residues 228 – 246 and the constituent amino acids of the active site are Asp44, Glu84 and Glu172. The enzyme belongs to Glycosyl Hydrolase/GH1 and GH5 family.

Impaired glucose balance affects the regeneration of neurotransmitters in the brain. One of neurotransmitters that is known to have an important role in the brain is dopamine. Dopamine production by dopaminergic (DA) neurons is regulated by the enzyme tyrosine hydroxylase (TH). Previous research has been reported that a decline on TH expression in DA neurons was a result of exposure to excess glucose. One contributing factor is an excess amount of glucose, which interfere homeostasis in the body by producing reactive oxygen species (ROS) more and cause oxidative stress. This condition can cause damage to cells in the brain, including DA neurons, to cell death occurs (apoptosis). The characteristic of neuroprotective owned by the hormone estrogen allegedly able to protect nerve cells from the damaging effects of this. The aim of this study is to explore the involvement of 17β-Estradiol as neuroprotection on the effects of glucose exposure during early development on dopaminergic neurons. This study used zebrafish (Danio rerio) embryos as animal model. They were divided into 4 groups: negative control group, hyperglycemia group, group that was coincubated glucose and estrogen, and the group that only incubated with estrogen alone. Cell death (apoptosis) in the brain was observed using acridine orange staining and TH expression was observed using immunocytochemistry. The results showed that the embryos were exposed to 5% glucose significantly (p less than 0.05) increased the number of apoptotic cells and decreased the expression of TH. While the results of the analysis of embryos in co-incubation group with estrogen showed improvement with decreased apoptotic cells and the increasing expression of TH up to the normal amount. It can be concluded that estrogen shown to have neuroprotective effect in protecting the decreasing TH expression, as the effects of glucose exposure, which could potentially lead to diabetes, by acting as an antioxidant through cellular mechanisms in the central nervous system

Several studies have been reported behavioral changes in patients with type 1 diabetes mellitus (T1DM) recently. The increasing levels of glucose affects neuronal activity and may inhibit the process of differentiation and regeneration of neurons in the brain. Dopaminergic (DA) neurons produce dopamine as a neurotransmitter which has an important role to regulate motor functions in the brain. This study aimed to determine the effect of exposure to high concentrations of glucose on the activity of dopaminergic neurons on early development of zebrafish (Danio rerio). Zebrafish embryos were exposed to glucose (1%, 3% and 5%) as a diabetic animal model at the beginning of development. Co-incubation was performed by incubated both of glucose and L-DOPA. Swimming distance of zebrafish larvae was measured as a parameter to determine the motor function regulated by dopamine activity. Results of statistical analysis in zebrafish embryos exposed to 1% and 3% glucose showed significant differences (p less than 0.05) reduction on swimming distance. Co-incubation glucose with L-DOPA partially increased the swimming distance. It can be concluded that the administration of excessive exposure to glucose was able to decrease the activity of dopaminergic neurons by decreasing the distance of swim in early zebrafish development. .

Summary: Humans develop anti-salivary proteins after arthropod bites or exposure to insect salivary proteins. This reaction indicates that vector bites have a positive effect on the host immune response, which can be used as epidemiological markers of exposure to the vector. Our previous study identified two immunogenic proteins with molecular weights of 31 kDa and 56 kDa from salivary gland extract (SGE) of Aedes aegypti that cross-reacted with serum samples from Dengue Hemorrhagic Fever (DHF) patients and healthy people in an endemic area (Indonesia). Serum samples from individuals living in non-endemic area (sub-tropical country) and infants did not show the immunogenic reactions. The objective of this research was to identify two immunogenic proteins, i.e., 31 and 56 kDa by using proteomic analysis. In this study, proteomic analysis resulted in identification of 13 proteins and 7 proteins from the 31 kDa- and 56 kDa-immunogenic protein bands, respectively. Among those proteins, the D7 protein (Arthropode Odorant-Binding Protein, AOBP) was the most abundant in 31-kDa band, and apyrase was the major protein of the 56-kDa band. Keywords: Proteomic analysis, Immunogenic proteins, Salivary gland, Aedes aegypti

Aggregatibacter actinomycetemcomitant (Aa) is gram-negative bacteria composed of lipopolysaccharides (LPS) in its outer membrane. This bacteria is normally found in dental plaque and periodontal pockets causing parotid gland damage when it is present in excess amounts. LPS is an endotoxin and a complex glycolipid composed of polysaccharides (hydrophilic) and lipid A (hydrophobic) which can induces Reactive Oxygen Species (ROS), and increase the levels of malonaldialdehyde (MDA) and tissue damage. The aims of this research are to find the expression of iNOS parotid gland cell and to determine levels of MDA using thiobarbituric acid (TBA) spectrophotometric test at 533 nm. The results showed a significantly difference (PAa rats (6.307 ± 0.0473 ppm). iNOS expression is confirmed by the brown spots formed on the parotid gland, affording the enhancements of iNOS expression in Aa rats and control rats of 2.292 ± 0.038 and 0.166 ± 0.003, respectively (P Keywords : Aggregatibacter actinomycetemcomitans, Reactive Oxygen Species, malondialdehyde, iNOS

Kesum (Polygonum minus L.) is a popular plant and is often used as additives in many typical dishes in the West Kalimantan. These plants belong to the family Polygonaceae, and potential to use in the prevention and treatment of lung cancer. This research aimed at studying the effect of giving the n-hexane fraction of the kesum to induces of apoptosis in epithelial cells of lung tissue of the animal models that exposed by benzopyrene. research was conducted on three groups of animal models: i) the healthy group, ii) the group treated by benzopyrene 200 mg/kg, and iii) the therapy group by n-hexane fraction 100 mg/kg. The results of the study then carried measurements of lung epithelial cell apoptosis index and measurements of lung ROS levels. The results showed that n-hexane fraction able to reduce of lung ROS levels of the animal models that exposed by benzopyrene with MDA levels respectively are 5.09 ± 0.76 ppm for the healthy group, 8.44 ± 1.89 ppm for the group treated by benzopyrene, and 5.47 ± 1.76 ppm for the therapy group. The n-hexane fraction also able to induce apoptosis of the lung epithelial cells of the animal models that exposed by benzopyrene with apoptosis index values respectively are 2.61 ± 1.98 for the healthy group, 7.41 ± 2.26 for the group treated by benzopyrene, and 16.31 ± 5.73 for the therapy group.

Curcumin is an agent affecting epigenetic factors and the expression of drug efflux transporters such as Cdr1p. Drug efflux transporters are also affected by the epigenetic factor HAT (Histone Acetyl Transferase). HAT-Rtt109 is specific to the fungal kingdom and related to resistance to antifungal agents. Strains of the pathogenic yeast Candida albicans were isolated from AIDS patients with oropharyngeal candidiasis and assigned to control and treatment groups (fluconazole, curcumin and the combination of fluconazole + curcumin). The minimum inhibitory concentration (MIC) for C. albicans resistant strains were determined using tube dilution test in accordance with CLSI micro dilution method, followed by SDS-PAGE. Yeast DNA isolation was performed using DNA isolation kit followed by bisulfate treatment, PCR, and sequencing of Rtt109. The most significant decrease in C. albicans growth occurred on growth media treated with the combination of fluconazole and curcumin. The MIC of the combination group was lower compared to the fluconazole-only or curcuminonly groups. Protein analysis showed that Cdr1p expression for the curcumin and combination groups was decreased compared to the control and fluconazole groups. There was a similar expression of HAT-Rtt109 among four groups. Based on the alignment of Rtt109 sequences, it shows that there was similar methylation among the four groups. The synergy of curcumin and fluconazole to helped in fluconazole resistance was likely through inhibition of Cdr1p expression, but not HAT-Rtt109 or Rtt109 methylation.

Hypertension is an abnormality of cardiovascular condition, marked by high-blood pressure over the normal condition. One of the factors that causes hypertension is altered equilibrium of Renin-Angiotensin-Aldosteron system (RAAS). This condition, which commonly by genetic could causes excess production of Angiotensin II, which known as vasoconstrictor, carried out by Angiotensin Converting Enzyme (ACE). Angiotensin II could increases the body’s blood pressure, and stimulates the production of free radicals, causing hypertension and oxidative stress. One of the common medication for hypertensive patient are using ACE inhibitor pills. Consumption of commercial pills, had a negative effect on the patient, such as skin iritation, cough, allergic reaction, etc. This study was conducted to explore the potential peptide of bakasang extract isolated from fermented skipjack tuna (Katsuwonus pelamis L.) for antihypertensive therapy, based on the activity of protease and histopathology of cardiac tissue on hypertensive rats (Rattus norvegicus) induced by DOCA-salt. The rats were divided into five groups: (1) negative control group, (2) hypertensive control group, (3) hypertension with captopril therapy dose of 5 mg/kg of body weight (BW), (4) and (5) hypertension with bakasang’s peptide extract therapy dose of 200 mg/kgBW, and 300 mg/kgBW, respectively. The results showed that bakasang’s peptide extract dose of 200 mg/kgBW significantly (p less than 0.05) decrease protease activity and improve histopathology of cardiac tissue on hypertensive rats. Protease activity post-therapy decreased to be 39.27 % from positive hypertension group. It could be concluded that the peptide of bakasang’s peptide extract has antihypertension effect (ACE-inhibitors), which was able to improve histopathology and decrease protease activity of the cardiac tissue on hypertensive rats induced by DOCA-salt.

Polyalthia longifolia (family: Annonaceae) is widely planted to effectively reduces noise pollution. This plant spread in many countries including Indonesia and the Philippines. Alkaloids are the main active compounds other than terpenoids in P.longifolia and it has the potential to be anti-inflammatory. Each leaves considered have different active compounds because of the different geographic factor from each country. To confirm this, we investigated the differences alkaloid compounds from two different sources of plants and predicted their anti-inflammatory potential. Shade-dried leaves from Indonesia and the Philippines were extracted by ethanol 70%. Two extracts were analyzed with LC-MS to ensure alkaloid compounds. Ensured alkaloid compounds further take on molecular docking. The compounds were drawn with ChemDraw then convert to .pdb with Open Babel. The protein COX-2 obtained from .pdb then prepared with PyMol. The docking process held by PyRx and the interaction was visualized by LigPlot+. LC-MS analysis identified 5 alkaloids contained from the ethanol extract of P.longifolia leaves from Indonesia and the Philippines. O-methylbulbocapnine-N-oxide was found only in ethanol extract leaves from Indonesia, while N-methylnandingerine-β-N-oxide was only found in ethanol extract leaves from the Philippines. All compounds have the potential as an anti-inflammation. Liriodenine as the most potent compound with binding energy −10.9kcal/mol. O-methylbulbocapnine-N-oxide has lower binding energy than N-methylnandingerine-β-N-oxide. In conclusion, there are differences between the alkaloid compounds and anti-inflammatory potential of the ethanol extract of P.longifolia leaves from Indonesia and the Philippines. Moreover, Indonesia’s ethanol extract leaves showed more potential than Philippines’s.

In the present study, phytochemical screening of ethanolic extracts of Azadirachta indica leaves from Indonesia and the Philippines revealed the presence of different phytoconstituents. Preliminary qualitative chemical test, TLC and LC-MS were used. TLC for all the extracts showed bands in long UV 366 for presence of flavonoid, tannin, saponin, terpenoid. As a result of LC-MS analysis of ethanolic extract Azadirachta indica leaves from Indonesia and Philippines, 10 compounds from Indonesian varian and 7 compounds from the Philippines varian were detected using m/z value. In conclusion, phytochemical screening based on TLC and LC-MS/MS show diverse bioactive compounds in ethanolic extract Azadirachta indica leaves from Indonesia and the Philippines. These can be effective approach for selecting best quality of varian leaves and planting area.

Acan is a typical fermented product from Maduranese, Indonesia. The product made from small shrimp called odhang acan or geraguh (Acetes sp.). Acan is a traditional condiment and has a distinctive aroma and flavor. This study aim is to identify the characteristics of Maduranese acan including raw material, fishing gears, manufacturing process, physical properties, chemical, and microbial profiles of the product. This research conducted by survey and interview technique, following by laboratory checking of the quality profile of the product. The results of the study indicate that acan produced by SMEs in Madura. The production of acan is quite simple through fermentation technology which added with 5-10% solar salt. Product properties have different characteristics depended on processing steps and fermentation period. Generally, the quality of acan has a convenient to the Indonesian shrimp paste standard product (SNI).

The main energy source commonly used by the community comes from fossil energy, especially petroleum. The energy crisis shows that Indonesia's fossil energy reserves are limited. Based on this fact, it is important to develop alternative energy that is environmentally friendly and sustainable, for example bioethanol. The purpose of this research is to screening and identify indegenous yeast from neera siwalan which is potential for bioethanol production. Isolation of yeast from neera siwalan is done using coconut medium. Purification is done to get one pure isolate. Screening is done by observing several parameters: the level of bioethanol production (using an alcohol meter), the content of reducing sugar with the DNS method, and the number of cells with optical density (600nm). Identification is done using ITS 4 and ITS 5. Based on the results of screening, four yeast isolates were produced, namely S3D, S1A, S2D, S1C. Based on these results the superior isolates producing bioethanol are S1A (14.4%). The results of the identification showed that the isolate was similar to Candida tropicalis with 100% similarity value.

Endophytic fungi are those fungi that are able to grow in plant tissue without causing symptoms of disease. It is thought that these fungi may confer on the host plants degree of resistance to parasitic invasion. Endophytic fungi have been isolated from stem tissue and these fungi are known to be antagonistic to pathogenic fungi. These endophytes produce chitinase and β-1,3-glucanase enzymes. Based on the fact that chitin and β-1,3-glucan are the main skeletal polysaccharides of the cell walls of fungal patogen. The aim of this research is to do potential test on some of isolates of Trichoderma's endophytic (L-1, L-2, Is-1, Is-2 and Is-7) in the chitinase and β-1,3-glucanase activity in effort to determine endophytic which be chossen to be gene resource for the next research. The gene will be transformed to citrus plant japanese citroen in effort to make citrus plant transgenic resistance to phytopatogenic invasion. The result of this research is endofit namely L-1 is the most potential endophytic fungi with chitinase activities is 4,8 10−2 Unit and glucanase 24,2. 1012 Unit. The addition of chitin and cell wall of phytophtora causes chitinase activity significantly increase, and also addition of laminarin and cell wall of phytophtora makes glucanase activity increase.

The aim of this research is to study antidiabetic activity of root extract of Ruellia tuberosa L on rats (Rattus novergicus) induced by multiple-low dose streptozotocin as animal diabetic models. The parameters investigated were blood glucose levels, free radicals (MDA, malondialdehyde) levels and hepatic histopathology. The main materials used were n-hexane root extracts from Ruellia tuberosa L. Three groups of rats, including control group (group I), diabetic group (group II), and therapy group with Ruellia tuberosa L (group III), were used. Streptozotocin was given at multiple-low dose of 20 mg/kg of body weight for 5 times in 5 consecutive days i.p. to rats in groups II and III. The Ruellia tuberosa L extracts were then given orally for group III in the dose of 250 mg/kg of body weight per day for 3 weeks. Results of the current work showed that root extract Ruellia tuberosa L had lowered blood glucose levels on rats in group III by 60.3%, from 299.7 ± 24.7 mg/dL up to 119.0 ± 26.6 mg/dL. Moreover, the antidiabetic activity of Ruellia tuberosa L extracts also deduced from decrease of MDA levels in group III, from 3.5 ± 0.3 μg/mL up to 1.7 ± 0.4 μg/mL. The recovery of hepatic organ from treatment group has also been proven from the its histology profiles stained with hematoxylin-eosin.

Induction laserpuncture effectively stimulates gonad maturation and spawning, but the associated gonadotropin releasing hormone (LH / GtH-II) mechanism is still unknown. So the purpose of this study demonstrate the mechanism of the release of GTH-II on catfish (Clarias sp.) after induction laserpuncture at the reproduction acupoint. The test fish consisted of 54 males and 54 females aged 8-9 months from the F1 hybrid Sangkuriang type female and Paiton type male parents. Ourstudy employed an experimental method with completely randomized design. The treatment comprised 6 levels with 9 repetitions. We were to observe gonadotropin hormone (GtH-II) concentrations pre-spawn, spawn, post-spawning in the laserpuncture exposed. As a comparison investigations were also conducted to untreated (control). Blood sampling performed at 6 h after induction laserpuncture. Hormone levels using the Elisa test kits Fish Luteinizing Hormone (LH) BIOTECH CO. CUSABIO., LTD. by Catalog No.. CSB-E15791Fh. The test results significant effect (P

The present study aimed to investigate the role of inhibin in the regulation of oocytesmaturation. Inhibin was isolated from goat granulose cells and injected by intraperitoneal in to femaleNile-Tilapia. Three days later its gonad was taken by decapitation and the change of oocyte was evaluatedunder microscope. Results showed that average of percentages of oocytes that undergo maturated after exposed with inhibin of 0, 20, 40, and 60µg/individual 50.05, 29.44, 21.46, and 30.83%, respectively.The exposure of doses of goat inhibin in Nile-Tilapia showed significantly different (P0.01) to percentages of mature oocytes. It was conluded that exposition of goat inhibin from granulose cells caninducing the maturation of Nile-Tilapia oocytes.Keywords: inhibin, maturation, oocytes

This paper reports a screening results of the secondary metabolites composed in Spathodea campanulata Beauv stem bark, evaluate inhibiting activity of malondialdehyde (MDA) on rat’s cancer model exposed with benzopyrene, and the histology of its lung. The secondary metabolite of the stem bark fraction consisted of alkaloids, flavonoids-phenolic, terpenoid and steroid compounds. The isolated fraction contained of these metabolites significantly indicate bioactivity by reducting of malondialdehyde (MDA) level, and also histology appearance of the lung tissue prepared from the benzopyrene-exposed rat indicated a curative activity.

Seagrass Thalassia hemprichii is used to study lead (Pb) metal accumulation and synthesis of phytochelatin (PC) and glutathione (GSH) as defense mechanisms against lead toxicity. The plants were exposed by lead (Pb (NO 3 ) 2 ) metal in some concentrations (0, 5, 15 and 25 ppm) for some periods of time (1, 2, 3 and 4 weeks). The contents of lead (Pb), phytochelatin (PC) and glutathione (GSH) are analyzed in leaf and root tissue. Lead accumulation in seagrass depends on the greatness of metal concentration and length of exposure time where the root accumulates lead higher than the leaf. Glutathione is produced higher in the root during lead exposure, while phytochelatin is produced in week 1 and 2. Then, production of phytochelatin turns to higher in the leaf in week 3, although it decreases in week 4. Phytochelatin and glutathione are formed at different retention time in UPLC, when glutathione content is formed earlier and lower than phytochelatin. The study not only reveals that content formation of phytochelatin (PC) and glutathione (GSH) occurs to respond Pb metal, but also shows that production of PC and GSH correlates significantly to Pb metal accumulation in the root and leaf of T. hemprichii . Therefore, in short, formation of PC and GSH, for sure, is a part of defense mechanisms conducted by seagrass T. hemprichii to protect itself against Pb metal toxicity.

Bakasang is a traditional fermented fish product which is often used as condiments. This study aimed to determine the physical and chemical characteristics and potential ACE (Angiotensin Converting Enzyme) inhibitor of bakasang. Color and viscosity were physical characteristics measured in which color was presented in value of L* (36.05), a* (18.76), b* (15.65), while viscosity value was 6 950 cP. The result of chemical characteristics including salinity, acidity, pH, TVB-N and LAB were 72 %, 2.56 %, 4.66, 36.88 mg N per 100 g and 3.32 log CFU g–1 respectively. Proximate and amino acid compositions analysis were also identified, resulting in 14.77 % protein, 1.11 % fat, 57.15 % moisture, 25.97 % ash and 1.00 % carbohydrate while the predominant amino acid found was histidine. The ACE inhibitory activity of the isolated bioactive peptides of bakasang was 68.80 %.