Diagnosis and Management of Hepatic Hydrothorax / 대한소화기학회지

Hepatic hydrothorax is a pleural effusion (typically ≥500 mL) that develops in patients with cirrhosis and/or portal hypertension in the absence of other causes. In most cases, hepatic hydrothorax is seen in patients with ascites. However, ascites is not always found at diagnosis and is not clinically detected in 20% of patients with hepatic hydrothorax. Some patients have no symptoms and incidental findings on radiologic examination lead to the diagnosis of the condition. In the majority of cases, the patients present with symptoms such as dyspnea at rest, cough, nausea, and pleuritic chest pain. The diagnosis of hepatic hydrothorax is based on clinical manifestations, radiological features, and thoracocentesis to exclude other etiologies such as infection (parapneumonic effusion, tuberculosis), malignancy (lymphoma, adenocarcinoma) and chylothorax. The management strategy involves a stepwise approach of one or more of the following: Reducing ascitic fluid production, preventing fluid transfer to the pleural space, fluid drainage from the pleural cavity, pleurodesis (obliteration of the pleural cavity), and liver transplantation. The complications of hepatic hydrothorax are associated with significant morbidity and mortality. The complication that causes the highest morbidity and mortality is spontaneous bacterial empyema (also called spontaneous bacterial pleuritis).

Detection mecA gene and Staphylococcus aureus resistance to several antibiotics isolated from cat ear swabs at a veterinary hospital located at Surabaya ­ Indonesia / Detecção do gene mecA e resistência a diversos antibióticos em Staphylococcus aureus isolados do ouvido de gatos em um hospital localizado em Surabaya ­ Indonesia

Cats are susceptible to S. aureus, which mainly colonizes the nose and ears of these feline species. Otitis externa in cat ears is one of the illnesses produced by S. aureus in animals. Antibiotic therapy for affected animals is the conventional treatment for infections by S. aureus. Antibiotic use during prolonged treatment and given at the wrong doses can cause germs to become resistant. Given this context, research on S. aureus isolated from cat ears and tests for antibiotic resistance and the mecA gene is required. Samples of cat ears were obtained from the Amies media using a sterile cotton swab. Bacterial isolation was done on MSA media, and then the catalase and coagulase assays were used to identify the bacteria. S. aureus isolates were evaluated for sensitivity using disks of the antibiotics cefoxitin, tetracycline, erythromycin, gentamicin, and chloramphenicol connected to MHA media. All positive isolates of S. aureus underwent MRSA testing, and then the mecA gene was detected. The sample investigation revealed that 91% (91/100) were positive for S. aureus, and 3.30% (3/91) were confirmed to be multidrug-resistant (MDR) because they are resistant to 3­4 antibiotic classes. Out of the 12 MRSA isolates analyzed, the mecA gene was detected in one isolate. Inappropriate antibiotic use causes bacterial resistance in pets. Additionally, excessive antibiotic use in a population might develop acquired bacterial resistance to an antibiotic. Antibiotic use in animals must be assessed to administer medication and prevent the development of antibiotic resistance appropriately.(AU)

Gatos são suscetíveis a adquirir S.aureus que colonizam principalmente as narinas e os ouvidos de espécies de felinos. A otite externa no ouvido dos gatos é uma das doenças produzidas pelo S.aureus nos animais. A terapia com antibióticos é o tratamento convencional para as infecções produzidas pelo S.aureus. Os antibióticos utilizados durante o prolongado tratamento e o emprego de sub doses podem selecionar microorganismos resistentes. Com base em tais argumentos torna-se necessária a pesquisa de S.aureus isolados do ouvido dos gatos, bem como, a realização de testes para a resistência a antibióticos e do gene mecA. Empregando swabs estéreis de algodão foram obtidas amostras dos ouvidos dos gatos em meio de Amies. O isolamento bacteriano foi efetuado em meio MAS e os testes catalase e coagulase foram realizados para a identificação das bactérias. A sensibilidade dos isolados de S.aureus foi avaliada com o emprego de discos dos antibióticos cefoxitin, tetraxiclina, eritromicina, gentamicina e cloranfenicol, incorporados no meio MHA. Todos os isolados positivos de S.aureus foram submetidos ao test MRSA para a detecção do gene mecA. A amostra investigada revelou 91% (91/100) de positivos para S.aureus, dos quais, 3,30% (3/91) foram resistentes a múltiplas drogas (MDR) pois foram resistentes a 3-4 classes de antibióticos. De 12 MRSA isolados analisados o gene mecA foi detectado em um isolado. O uso inapropriado de antibióticos é a causa da resistência bacteriana em pets. Adicionalmente o emprego excessivo de antibióticos em uma população pode resultar no desenvolvimento de resistência bacteriana adquirida a antibióticos. O uso de antibióticos em animais deve ser ordenado por uma administração de medicamentos apropriada para prevenir o desenvolvimento da resistência.(AU)

Effect of Spirulina on Cochlea Histopathological Changes in Wistar Rats Induced by Kanamicin

@#Introduction: Streptomycin and kanamycin are aminoglycosides that are toxic to the cochlea vestibular system, can causing hearing loss. This antibiotic is used for the treatment of tuberculosis and its ototoxicity occurs in 20% of tuberculosis patients. Spirulina is a cyanobacterial species that is used as a dietary supplement and contains phycocyanin compounds that function as antioxidants and anti-inflammatory. The aim of this study was to determine the effect of spirulina on histopathological changes in the cochlea in Wistar rats after kanamycin induction. Methods: this study is a form of posttest-only controlled group design research with a sample of 24 wistar rats divided into 4 groups, namely negative control group, positive control group, treatment group 1 and treatment group 2. Observations of the study took place in November-December 2021. Histopathological measurements in hair cells, macrophages and cochlear vasculature. The analysis used non-parametric Kruskal-Wallis and post-hoc Mann-Whitney tests. Results: There were more hair cell damage, macrophage cell count, and significant vascular dilatation in the kanamycin group than in the without kanamycin group with the value p=0.001. There was significantly less number of hair cell damage in the kanamycin group with spirulina at a dose of 1000 mg than in the kanamycin group with spirulina at a dose of 400 mg p=0.045. Conclusion: There was a significant effect on the administration of spirulina on histopathological changes in the cochlea of rats.

Fatty Liver in Hormone ReceptorPositive Breast Cancer and Its Impact on Patient's Survival / 한국유방암학회지

Purpose@#Long-term estrogen inhibition may cause fatty liver disease (non-alcoholic fatty liver disease; NAFLD) among other adverse conditions such as osteoporosis, climacteric symptoms, thromboembolism, dyslipidemia, and metabolic syndrome. The prevalence of NAFLD among breast cancer patients ranges from 2.3%–45.2%. This study aimed to determine the risk factors for newly developed NAFLD among breast cancer patients after hormonal treatment and whether it influences survival outcomes. @*Methods@#This retrospective study investigated hormone receptor (HR)-positive, human epidermal growth factor receptor 2 (HER2)-negative (HR+/HER2−), nonmetastatic breast cancer patients diagnosed between January 2010 and December 2018. All patients received adjuvant hormonal treatment for at least 6 months. Clinical data on metabolic profile indicators such as body mass index (BMI), waist circumference, serum cholesterol, triglycerides, low-density lipoprotein-cholesterol (LDL-C), high-density lipoproteincholesterol (HDL-C), diabetes, and presence of metabolic syndrome (MetS) were collected. In total, 160 eligible patients with complete covariate data and survival follow-up were included. @*Results@#NAFLD was diagnosed in 35% of patients. There were significant associations of being overweight (BMI ≥ 25 kg/m2 ), waist circumference > 80 cm, triglycerides ≥ 150 mg/ dL, HDL-C ≤ 50 mg/dL, LDL-C < 150 mg/dL, and presence of MetS with the development of NAFLD. However, unlike other factors, MetS and HDL-C were not independently associated with NAFLD. Patients with breast cancer who developed NAFLD had longer disease-free survival (DFS). The median DFS was not reached in the NAFLD group, whereas it was 59.3 (45.6–73.0) months in the non-NAFLD group. No worsening of overall survival was observed in patients with breast cancer and NAFLD. @*Conclusion@#The development of NAFLD during treatment in patients with HR+/HER2− breast cancer was associated with several independent risk factors: being overweight, waist circumference, triglycerides, and LDL-C. Interestingly, breast cancer patients with NAFLD during treatment had longer DFS than those without NAFLD.

Perspectives on sustainable management of the Poso Lake (Indonesia) endemic ricefish, Oryzias nigrimas (Actinopterygii: Adrianichthyidae) / Perspectivas sobre el manejo sostenible del pez Oryzias nigrimas, endémico del Lago Poso en Indonesia (Actinopterygii: Adrianichthyidae)

Introduction: The endemic fishes of the ancient lakes of Sulawesi are under increasing threat. Objective: To evaluate the data and information available from a holistic management perspective and to formulate measures to conserve the endemic ricefish Oryzias nigrimas in Poso Lake, Indonesia. Methods: Collection of primary data from three stations around Lake Poso and literature study. Results: Threats to O. nigrimas include habitat degradation and loss, introduced alien species, and exploitation as a locally important food fish. Options to promote sustainable fisheries management include spatial and temporal limitations to minimise catch of gravid or brooding fish. Habitat protection should include measures to minimise impacts from activities which can reduce water quality and disturb or kill aquatic vegetation. Conclusion: Measures to prevent further O. nigrimas population decline are considered urgent and further research is recommended to fill identified knowledge gaps. Ex-situ conservation, including the development of captive breeding, could also contribute to a holistic O. nigrimas conservation strategy.

Introducción: Los peces endémicos de los antiguos lagos de Sulawesi están cada vez más amenazados. Objetivo: Evaluar los datos y la información disponible desde una perspectiva de gestión holística y formular medidas para conservar el pez endémico Oryzias nigrimas en el lago Poso, Indonesia. Métodos: Recopilación de datos primarios de tres estaciones alrededor del lago Poso y revisión de la literatura. Resultados: Las amenazas para O. nigrimas incluyen la degradación y pérdida del hábitat, la introducción de especies exóticas y la explotación como producto pesquero de importancia local. Las opciones para promover la ordenación pesquera sostenible incluyen limitaciones espaciales y temporales para minimizar la captura de peces grávidos o reproductores. La protección del hábitat debe incluir medidas para minimizar los impactos de las actividades que pueden reducir la calidad del agua y perturbar o matar la vegetación acuática. Conclusión: Las medidas para prevenir una disminución de la población de O. nigrimas se consideran urgentes y se recomienda realizar más investigaciones para llenar los vacíos de conocimiento identificados. La conservación ex situ, incluido el desarrollo de la cría en cautividad, también podría contribuir a una estrategia de conservación holística de O. nigrimas.

Techniques Closed Suction Influence on Oxygen Saturation In Patients Using Mechanical Ventilation In Intensive Care Unit Room

@#Introduction: The problem in patients who are intubated with endotracheal tube is the occurrence of accumulation of secretions. One of the actions to reduce the secretion is closed suction. This research monitors the patient's oxygensaturation and also reveals the effect of closed suction on oxygen and AaDO2 saturation (alveolar-arterial oxygen difference) in patients who were mechanically ventilated in the Intensive Care Unit (ICU) of Dr. Kariadi Hospital. The design of this study uses pre experiment. Methods: The research process has been carried out on April 2, 2019 until May 26, 2019 in the Intensive Care Unit with a total sampling technique, on 15 respondents. Result: The results of the study showed that there was effect of closed suction technique on the value of oxygen saturation in patients with mechanical ventilation (p-value = 0.010). Conclusion: Closed suction can lead to decrease of air volume in the lungs up to 1200 ml. So hyperoxygenation must be done before closed suction as it can reduces the incidence of hypoxemia by 49%. Before closing the suction, it is recommended to do hyperoxygenation.

A novel wound healing accelerator: Effect of vitreous gel of cow eyeball on a chronic wound model

@#ss of wound healing after administration ofmitomycin-C, which inhibits granulation tissue formationand collagen synthesis, resulting in chronic wounds. Thevitreous gel of cow eyeballs contains a high level ofhyaluronic acid, which has a role in inflammation,granulation, re-epithelialization, and remodelling. This studyaims to understand the effect of 1% povidone iodine andvitreous gel of cow eyeballs on wound healing afteradministration of mitomycin-C.Methods: This was an invivostudy with quasi-experimentalmethods on 32 Wistar mice. Full-thickness wounds weremade and then treated with mitomicyn-C. The mice weredivided into 4 groups: a control group with NaCl 0.9%vitreous gel of cow eyeball (VGCE), 1% povidone-iodine, anda combination of VGCE and 1% povidone-iodine groups.Macroscopic and microscopic observations of the processof wound healing were performed on days 3, 7, and 14.Results: Vitreous gel administration produced significantwound healing rates within the first three days, andhistological analysis revealed an increased number offibroblasts and polymorphonuclear cells. However, thepovidone iodine group and the combination group withvitreous gel did not produce significant results.Conclusion: The single administration of VGCE canaccelerate the wound healing process, increase the numberof fibroblasts, and reduce inflammation in a chronic woundmodel.

Antioxidant activity, total phenolic and flavonoid contents of selected medicinal plants used for liver diseases and its classification with chemometrics

Liver disease has become one of the major health problems in the world, and the death rate is going rapidly toincrease. Oxidative stress plays a crucial role in the emerging, development, and the progression of liver diseases.Ethnobotanical research has an undoubted profound impact on the development of numerous new drugs. The aimof this research, therefore, was to examine the antioxidant activities of 14 selected plants used for treating liverdiseases by traditional healers of Indonesia ethnicities and to classify these plants using chemometrics of principalcomponent analysis (PCA). The extraction using methanol as the solvent was performed with two stages maceration.Total phenolic and flavonoid compounds were determined by Folin–Ciocalteau and AlCl3 method, respectively,whereas antioxidant activity was estimated using 2,2′-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging,trolox equivalent antioxidant capacity (TEAC), and ferric reducing antioxidant power (FRAP) assay. Among 19methanol extracts of 14 plants, the leaves of Baccaurea racemosa, Macaranga subpeltata, and Piper sp. showed thehighest antioxidant properties. The phenolic content correlated with TEAC, FRAP, and DPPH radical scavengingactivity, while flavonoid did not significantly affect these antioxidant activity methods. PCA successfully classifiedthe plant samples using the variables of antioxidant activities and phenolic-flavonoid contents. The selected plantshave promising antioxidant properties which support their utilization for either liver diseases medication or oxidativestress-related diseases prevention

Dosage-Related Prebiotic Effects of Inulin in Formula-Fed Infants / 대한소아소화기영양학회지

PURPOSE: The aim of this study was to identify the minimally meaningful dosage of inulin leading to a prebiotic effect in Indonesian infants. METHODS: In a randomized controlled double-blinded, parallel, 3-arm intervention study, 164 healthy formula-fed infants aged 3 to 5 months first obtained formula-A (without inulin) during a 4-week adaptation period. Subsequently, 142 subjects were subjected to a 4-week feeding period by administering either formula-A (no inulin), formula-B (0.2 g/100 mL inulin) or formula-C (0.4 g/100 mL inulin). The primary outcome parameter was %-bifidobacteria in faecal samples determined using quantitative polymerase chain reaction analyses. Secondary outcome parameters were faecal %-lactobacilli, pH and stool frequency, and consistency. Growth and tolerance/adverse effects were recorded as safety parameters. RESULTS: Typical %-bifidobacteria and %-lactobacilli at the end of the adaptation period in the study population were 14% and 2%, respectively. For faecal pH, significant differences between formula groups A vs. C and A vs. B were found at the end of the intervention period. Testing for differences in faecal %-bifidobacteria and %-lactobacilli between groups was hampered by non-normal data set distributions; no statistically significant differences were obtained. Comparisons within groups revealed that only in formula group C, all the three relevant parameters exhibited a significant effect with an increase in faecal %-bifidobacteria and %-lactobacilli and a decrease in pH. CONCLUSION: A consistent prebiotic effect along with a decrease in pH and increase in %-bifidobacteria and %-lactobacilli was found only in the group administered 0.4 g inulin/100 mL.

The Role of Two Human Milk Oligosaccharides, 2′-Fucosyllactose and Lacto-N-Neotetraose, in Infant Nutrition / 대한소아소화기영양학회지

Human breast milk contains numerous biomolecules. Human milk oligosaccharides (HMOs) are the third most abundant component of breast milk, after lactose and lipids. Amongst the synthetized HMOs, 2′-fucosyllactose (2′-FL) and lacto-N-neotetraose (LNnT) are widely studied and are considered safe for infant nutrition. Several studies have reported the health benefits of HMOs, which include modulation of the intestinal microbiota, anti-adhesive effect against pathogens, modulation of the intestinal epithelial cell response, and development of the immune system. The amount and diversity of HMOs are determined by the genetic background of the mothers (HMO secretors or non-secretors). The non-secretor mothers secrete lower HMOs than secretor mothers. The breastfed infants of secretor mothers gain more health benefit than those of non-secretor mothers. In conclusion, supplementation of infant formula with 2′-FL and LNnT is a promising innovation for infant nutrition.

V279F polymorphism might change protein character and immunogenicity in Lp-PLA2 protein

Background: Lipoprotein-associated phospholipase A2 [Lp-PLA2] plays a strong role in the occurrence of certain cardiovascular disease processes. Polymorphisms at the protein level are also estimated to correlate with increased risk factors for heart attacks. One such polymorphism is the V279F polymorphism in Lp-PLA2 which results in a change in enzyme performance capability. This in turn implies a reduced risk of acute myocardial infarct [AMI] in Korean and Indonesian subpopulations

Aim: This study aimed to analyze changes in protein properties, structure, energy stability, epitope, and immunogenicity that are due to the substitution of the Valine [V] amino acid at position 279 to Phenylalanine [F] in the Lp-PLA2 protein

Methodology: The role of Lp-PLA2 in the cardiovascular process and in AMI was analyzed based on the protein-protein network according to the BioGRID, MENTHA, and STRING databases. Protein properties and energy stability were examined by FoldX; this was followed by identification of epitope using ElliPro. The immunogenicity was evaluated in vivo by injecting the protein into mice and subsequently measuring their antibody production using the ELISA technique

Results: The substitution of Valine for Phenylalanine was predicted to increase protein stability and epitope shifts. Further studies on animal experiments exhibit that the 279V variant is able to induce IgG production better than the 279F variant

Conclusion: Based on these data, it can be concluded that the V279F polymorphism influences the surface structure, energy stability, epitope and immunogenicity of the Lp-PLA2 protein. The changes in the immunogenicity and epitope shift indicated that the protein is valuable as a biomarker for use in acute myocardial infarct. The results of this study provide an opportunity to develop monoclonal antibodies that are specifically able to identify V279F polymorphisms as a predictor of the risk of acute myocardial infarct

Alu insertion/deletion of ACE gene polymorphism might not affect significantly the serum bradykinin level in hypertensive patients taking ACE inhibitors

Background: Angiotensin I-converting enzyme [ACE] has two homologous catalytic domains, the N- and C-domains. Our previous study suggested that Alu insertion [I allele] in the intron 16 of ACE resulted in premature codon termination. The I allele has only one active site in the N-domain while the Alu deletion [D allele] still has two active sites of ACE. Therefore the effect of I/ D polymorphism of ACE on the enzyme's ability to catalyse bradykinin is still not widely known

Aims: This study aimed to examine the serum bradykinin level in hypertensive patients with I/D polymorphism of ACE, who were treated with ACE inhibitor

Subjects and methods: The serum bradykinin and I/D polymorphism have been detected in 64 hypertensive patients taking ACE inhibitor [lisinopril or captopril] for at least eight weeks with good medication adherence. The binding affinity of ACE with its receptor was calculated by molecular docking

Results: The findings show that genotype II is more frequent in the population the researchers observed [53.12%] compared to ID [23.44%] and DD [23.44%] variances. On the other hand, the bradykinin level is not affected by genotype of the ACE genes on the population. Bradykinin increases in patients with genotype II who are given captopril, but decreases in patients treated with lisinopril. Nevertheless, there is no statistically significant difference

Conclusion: This study suggests that the polymorphism might not significantly affect the serum bra-dykinin level in hypertensive patients taking ACE inhibitors

In silico molecular analysis of novel L-specific dehalogenase from Rhizobium sp. RC1

Aims: This study presents the first structural model and proposed the identity of four important key amino acid residues, Asp13, Arg51, Ser131 and Asp207 for the stereospecific haloalkanoic acid dehalogenase from Rhizobium sp. RC1. Methodology and results: The enzyme was built using a homology modeling technique; the structure of crystallized LDEX YL from Pseudomonas sp. strain YL as a template. Model validation was performed using PROCHECK to generate the Ramachandran plot. The results showed 80.4% of its residues were located in the most favoured regions suggested that the model is acceptable. Molecular dynamics simulation of the model protein was performed in water for 10 nanoseconds in which Na+ was added to neutralize the negative charge and achieved energy minimization. The energy value and RMSD fluctuation of Cα backbone of the model were computed and confirmed the stability of the model protein. Conclusion, significance and impact of study: In silico or computationally based function prediction is important to complement with future empirical approaches. L-haloacid dehalogenase (DehL), previously isolated from Rhizobium sp. RC1 was known to degrade halogenated environmental pollutants. However, its structure and functions are still unknown. This structural information of DehL provides insights for future work in the rational design of stereospecific haloalkanoic acid dehalogenases.

comparative analysis of serum albumin from different species to determine a natural source of albumin that might be useful for human therapy

Objectives: Serum albumin [SA] is one of the most abundant proteins in human plasma and performs functions for protein transport, scavenges for free radicals, and regulates osmotic pressure. SA has been used for therapy in humans with several diseases including hypovolemia, hypoalbuminemia, and chronic liver disease. However, the most appropriate source of albumin for medical applications remains unclear, especially in the case of viral infections, which complicates finding viable donors. Substitution of human serum albumin [HSA] with albumin from other natural sources is a viable alternative. Therefore, we elucidated the similarity in character, structure and evolutionary relationship among serum albumin isolated from seven different species

Methods: We compared the sequence, structure, and properties of SA from different species using an in-silico approach

Results: These data suggested that SA has sequence polymorphism that clusters based on closely relatedspecies. However, these polymorphisms do not change the three-dimensional structure of the protein; this may serve to maintain its function as a transporter. The Gallus gallus albumin has the lowest number of the epitopes that closely resemble HSA

Conclusion: This study is crucial in providing explicit information about the structural similarity of albumin isolated from other species compared to HSA. The Gallus gallus SA might be used as a primary natural source of albumin where warranted for human therapy

Identification of antigenic proteins from salivary glands of female Anopheles maculatus by proteomic analysis

Objective To identify antigenic proteins from the salivary glands of female Anopheles maculatus using a proteomic approach to find the biomarker candidate for serological tools. Methods The identification of antigenic proteins of Anopheles maculatus salivary gland used these techniques: one-dimensional gel electrophoresis (sodium dodecyl sulfate polyacrylamide gel electrophoresis), western blot, and liquid chromatography–mass spectrometry. Results The proteins that have molecular weight (MW) 43 and 34 kDa were the antigenic protein. Computational bioinformatic analysis by Mascot Server revealed seven novel hypothetical proteins (MW: 43 kDa) and two novel hypothetical proteins (MW: 34 kDa). Further analysis (BLASTP, antigenicity, epitope mapping, and specificity analysis) showed that two novel proteins were identified as apolipoprotein D and cathepsin D in Anopheles darlingi. Conclusions The identified proteins are potential to be developed as a biomarker of mosquito bite's exposure.

Platelet-Rich Fibrin Lysate Can Ameliorate Dysfunction of Chronically UVA-Irradiated Human Dermal Fibroblasts

To determine whether platelet-rich fibrin lysate (PRF-L) could restore the function of chronically ultraviolet-A (UVA)-irradiated human dermal fibroblasts (HDFs), we isolated and sub-cultured HDFs from six different human foreskins. HDFs were divided into two groups: those that received chronic UVA irradiation (total dosages of 10 J cm-2) and those that were not irradiated. We compared the proliferation rates, collagen deposition, and migration rates between the groups and between chronically UVA-irradiated HDFs in control and PRF-L-treated media. Our experiment showed that chronic UVA irradiation significantly decreased (p<0.05) the proliferation rates, migration rates, and collagen deposition of HDFs, compared to controls. Compared to control media, chronically UVA-irradiated HDFs in 50% PRF-L had significantly increased proliferation rates, migration rates, and collagen deposition (p<0.05), and the migration rates and collagen deposition of chronically UVA-irradiated HDFs in 50% PRF-L were equal to those of normal fibroblasts. Based on this experiment, we concluded that PRF-L is a good candidate material for treating UVA-induced photoaging of skin, although the best method for its clinical application remains to be determined.

Decreasing α-synuclein aggregation by methanolic extract of Centella asiatica in zebrafish Parkinson's model

Objective: To observe the effects of Centella asiatica ( C. asiatica) methanolic extract on α-synuclein aggregation and its expression in rotenone-exposed zebrafish. Methods: Zebrafish ( Danio rerio) were exposed to 5 μg/L rotenone for 28 days and co-incubated with 2.5, 5.0 and 10.0 μg/mL of C. asiatica methanolic extract. The medium was changed every 48 h for maintain the concentration of rotenone and extract. After 28 days zebrafish were sacrificed on the ice block and protein was isolated from zebrafish brain for ELISA of dopamine and Western blotting of α-synuclein. Immunohistochemistry was conducted to observe the α-synuclein expressions from histopathological preparation of zebrafish brain. The head were soaked in 10% formaline for less than 24 h and embedded onto paraffin block, then sliced for immunohistochemistry using anti α-synuclein antibody. We also measured zebrafish motility for 5 min in each week. Results: C. asiatica has important bioactive compounds such as asiaticoside that has anti-inflammatory and antioxidant properties. It may inhibit cascade reaction due to oxidative stress induced by rotenone. Decreasing reactive oxygen species proposed probability of radical attack to α-synuclein protein that caused aggregation and increase of its expression. The motility of zebrafish was also maintained in C. asiatica groups due to the increasing dopamine level in rotenone-induced zebrafish. High level of reactive oxygen species inactivated enzyme for dopamine synthesis such as tyrosine hydroxylase, and oxidized dopamine itself. Oxidized dopamine increased α-synuclein aggregation. Thus, the dopamine level decreased in rotenone-induced zebrafish, but C. asiatica increased dopamine level. Conclusions: C. asiatica has a potential to be developed as an anti-Parkinson's disease treatment due to its capability for minimized the sign of Parkinson's such as α-synuclein aggregation and expression, increasing motility and dopamine as well.

Anti-diabetic potential of Urena lobata leaf extract through inhibition of dipeptidyl peptidase IV activity

Objective: To evaluate the anti-diabetic potential of leaf extract from Urena lobata (U. lobata) through dipeptidyl peptidase IV (DPP-IV) inhibitory activity. Methods: U. lobata leaf was extracted in hot water and ethanol. The activity of DPP-IV inhibitor was tested by in vitro study using gly-pro-p-nitroanilide as substrat of DPP-IV and vildagliptin, as standard reference. A product of the reactions between gly-pro-pnitroanilide and DPP-IV, was observed by microplate readers with λ = 405 nm. All data were expressed as mean ± SD and the IC50 value was determined by non linear regression curve fit. Active substances in leaf extract of U. lobata was analyzed by liquid chromatography-mass spectrometry. DPP-IV inhibitory activity of active compounds was evaluated in silico using docking server. Results: The ethanolic extract of U. lobata showed stronger DPP-IV inhibitor activity than water extract with the IC50 values of 1 654.64 and 6 489.88 mg/mL, respectively. Vildagliptin, based on standard reference for DPP-IV inhibitor activity, has IC50 value of 57.44 μg/mL. Based on in silico analysis, mangiferin, stigmasterol and β-sitosterol in U. lobata extract have a strong inhibitory activity on DPP-IV. Conclusions: The results showed that DPP-IV inhibitory activity of U. lobata is related to its active compounds such as mangiferin, stigmasterol and β-sitosterol.

Plasma CRH Level Difference Between Wistar Rats Exposed To Acute Stress Due To Predator And To The Psychological Stress Device

Objective: Stress triggers and causes psychiatric disorders. This study compared stress generated by different stressors: a cat as the predator of rats and a Psychological Stress Device (PSD) which was developed and modified by the researchers based on the model by Xu and Rocher. Methods: Twenty-eight Wistar rats were simple randomly divided into one control group and six treatment groups, each consisting of 4 rats. Each treatment group was individually exposed to stressor for 30, 60, and 90 minutes. The first three treatment groups were treated using the PSD while the other three treatment groups were treated exposed to the cat. Plasma CRH level was measured using the ELISA (Cusabio) method. Result: Plasma CRH levels in the rat exposed to stressor using the PSD ranged from 9.89 to 50.22 ng/mL, higher than plasma CRH level in the groups exposed to cat ranged from 0.22 to 23.44 ng/mL with significance level (p0.05). Conclusion: Plasma CRH level of the rats exposed to stressor using the PSD was higher and positively correlate with the length of exposure compared to those exposed to cat.

Decreasingα-synuclein aggregation by methanolic extract of Centella asiatica in zebraifsh Parkinson’s model

Objective:To observe the effects of Centella asiatica (C. asiatica) methanolic extract onα-synuclein aggregation and its expression in rotenone-exposed zebrafish. Methods: Zebrafish (Danio rerio) were exposed to 5 μg/L rotenone for 28 days and co-incubated with 2.5, 5.0 and 10.0 μg/mL of C. asiatica methanolic extract. The medium was changed every 48 h for maintain the concentration of rotenone and extract. After 28 days zebrafish were sacrificed on the ice block and protein was isolated from zebrafish brain for ELISA of dopamine and Western blotting of α-synuclein. Immunohistochemistry was conducted to observe the α-synuclein expressions from histopathological preparation of zebrafish brain. The head were soaked in 10%formaline for less than 24 h and embedded onto paraffin block, then sliced for immunohistochemistry using antiα-synuclein antibody. We also measured zebrafish motility for 5 min in each week. Results:C. asiatica has important bioactive compounds such as asiaticoside that has anti-inflammatory and antioxidant properties. It may inhibit cascade reaction due to oxidative stress induced by rotenone. Decreasing reactive oxygen species proposed probability of radical attack toα-synuclein protein that caused aggregation and increase of its expression. The motility of zebrafish was also maintained in C. asiatica groups due to the increasing dopamine level in rotenone-induced zebrafish. High level of reactive oxygen species inactivated enzyme for dopamine synthesis such as tyrosine hydroxylase, and oxidized dopamine itself. Oxidized dopamine increasedα-synuclein aggregation. Thus, the dopamine level decreased in rotenone-induced zebrafish, but C. asiatica increased dopamine level. Conclusions: C. asiatica has a potential to be developed as an anti-Parkinson's disease treatment due to its capability for minimized the sign of Parkinson’s such asα-synuclein aggregation and expression, increasing motility and dopamine as well.