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Context: Acute myocardial infarction (AMI) is one of the clinical manifestations of coronary heart disease caused by the cessation of coronary blood flow. The development of the management of AMI has resulted in a reduction in the mortality rate of AMI patients. Nevertheless, the apparent inflammation triggers detrimental remodeling, so antifibrotic such as colchicine are needed. Aims: To analyze the impact of colchicine administration on the reduction of ventricular remodeling through the NLRP3 inflammasome, TGF-β, and α-SMA in a 3T3 cell line culture under ischemic conditions. Methods: The 3T3 cell line culture underwent ischemia treatment using CoCl2 and subsequent treatment with colchicine. This treatment was conducted for 24 hours with a concentration of 300 μM CoCl2, followed by the administration of colchicine at a concentration of 1 μM after the ischemia treatment. The expression levels of NLRP3, TGF-β, and α-SMA were assessed using flow cytometry, and the data were analyzed through one-way ANOVA. Results: Administration of colchicine for 24 hours following ischemia resulted in a significant decrease in the relative levels of NLRP3, TGF-β, and α-SMA (p

Background: Insomnia is difficulty initiating or maintaining sleep for at least three nights a week or more and lasting for at least 3 months. One of the molecules that play a role in the circadian rhythm of arousal system is hypocretin/orexin. Orexin activates the p38-MAPK signaling pathway and increases phosphorylated ERK1/2 levels. Centella asiatica (CA) has a role in the signal work of the MAPK/ERK, Akt, and p38 path in many various diseases. Methods: The research method used is true laboratory experimental. The research approach used was randomized control group post-test only. Zebrafish embryos aged 0-7 dpf were used in this study. The treatment group consisted of 5 groups: normal, insomnia, insomnia + 2.5 μg/mL CA, insomnia + 5 μg/mL CA, and insomnia + 10 μg/mL CA. The locomotor motion of zebrafish larvae was observed using Basler cameras on days five-, six- and seven-day post fertilization (dpf), then analyzed by using Western Blot method. Results: The results proved that exposure to CA extract was able to reduce the expression of orexin (91963 ± 9129) and p38 (117425 ± 6398) as an arousal trigger in the sleep-wake cycle, with the most optimal concentration of CA 5 μg/mL. Exposure to CA extract was also able to reduce the expression of ERK (94795 ± 30830) and Akt (60113.5 ± 27833.5) with an optimum concentration of CA 2.5 μg/mL. Conclusion: Exposure to CA extract was able to improve the sleep activity of zebrafish larvae insomnia model by extending the total inactivity time (cumulative duration) and shortening the duration of first sleep (latency to first) in light and dark phases through inhibition of orexin, ERK, p38, and Akt.

Introduction The main alkaloid component in cigarettes is nicotine. Cotinine, a metabolite of nicotine, is capable of causing dependence effects through endless mechanisms modulated by the ion channel nicotinic acetylcholine receptors nAChRs. Nicotine and cotinine can also cause damage to blood vessels through a chronic inflammatory process mediated by the Ligand-Tie2 Angiopoietin Receptor system. Hypoxic conditions that occur due to vascular inflammation cause a decrease in the concentration of nitric oxide (NO). This study aimed to evaluate the relationship between NO levels and cotinine through the expression of nAChRs that mediate the nicotine dependence mechanism and Tie2 (Tyrosine Kinase 2) expression. Methods A cross-sectional study was conducted with 200 participants grouped into two groups based on their smoking status: 100 smokers and 100 non-smokers. All participants were men aged 20–40 years with no history of cardiovascular disease, diabetes mellitus, or dyslipidemia, and were not currently on medication. According to the parameters used, all blood samples were taken from peripheral blood for analysis using the ELISA kit or Colorimetric Assay Kit. Results Cigarette consumption increases blood cotinine concentrations in smokers and causes dependence by modulating nAChRs. The study indicates an emerging cycle regarding nicotine–cotinine consumption and nAChRs expression. In addition, the data in this study showed a significant relationship (p

Abstract Background Inflammaging, the characteristics of immunosenescence, characterized by continuous chronic inflammation that could not be resolved. It is not only affect older people but can also occur in young individuals, especially those suffering from chronic inflammatory conditions such as autoimmune disease, malignancy, or chronic infection. This condition led to altered immune function and as consequent immune function is reduced. Detection of immunosenescence has been done by examining the immune risk profile (IRP), which uses flow cytometry. These tests are not always available in health facilities, especially in developing countries and require fresh whole blood samples. Therefore, it is necessary to find biomarkers that can be tested using stored serum to make it easier to refer to the examination. Here we proposed an insight for soluble biomarkers which represented immune cells activities and exhaustion, namely sCD163, sCD28, sCD80, and sCTLA-4. Those markers were reported to be elevated in chronic diseases that caused early aging and easily detected from serum samples using ELISA method, unlike IRP. Therefore, we conclude these soluble markers are beneficial to predict pathological condition of immunosenescence. Aim To identify soluble biomarkers that could replace IRP for detecting immunosenescence. Conclusion Soluble costimulatory molecule suchsCD163, sCD28, sCD80, and sCTLA-4 are potential biomarkers for detecting immunosenescence.

BACKGROUND: Colorectal cancer (CRC) is reported as the third most frequently diagnosed cancer worldwide. Salmonella infection plays a role in developing this cancer, which is chronically related to the avirulence protein A (AvrA) effector protein produced by the bacteria. This study was conducted to point out the effect of AvrA Salmonella on the occurrence of CRC through the regulation of TLR4/NF-κB/β-catenin/TGF-β expressions in the azoxymethane (AOM)/dextran sodium sulfate (DSS) CRC model. METHODS: A randomized control group post-test-only study was conducted using male Balb/c mice with 30-gram body weight (10-12 weeks), which were divided into three groups, namely the negative control (normal mice without any treatment), positive control (AOM/DSS-treated mice), treatment 1 (T1) (AOM/DSS-treated mice + AvrA Salmonella) groups. Colon tissue was collected and then prepared for immunohistochemistry staining using TLR4, β-catenin, NF-κB, TGF-β, and Ki67 antibodies, whereas apoptotic cells were stained using TUNEL assay. RESULTS: The expressions of TLR4, β-catenin, NF-κB, TGF-β, Ki67, and apoptosis percentage indicated significant differences among the three groups, which statistically showed p

Background: Insomnia is difficulty initiating or maintaining sleep for at least three nights a week or more and lasting for at least 3 months. One of the molecules that play a role in the circadian rhythm of arousal system is hypocretin/orexin. Orexin activates the p38-MAPK signaling pathway and increases phosphorylated ERK1/2 levels. Centella asiatica (CA) has a role in the signal work of the MAPK/ERK, Akt, and p38 path in many various diseases. Methods: The research method used is true laboratory experimental. The research approach used was randomized control group post-test only. Zebrafish embryos aged 0-7 dpf were used in this study. The treatment group consisted of 5 groups: normal, insomnia, insomnia + 2.5 μg/mL CA, insomnia + 5 μg/mL CA, and insomnia + 10 μg/mL CA. The locomotor motion of zebrafish larvae was observed using Basler cameras on days five-, six- and seven-day post fertilization (dpf), then analyzed by using Western Blot method. Results: The results proved that exposure to CA extract was able to reduce the expression of orexin (91963 ± 9129) and p38 (117425 ± 6398) as an arousal trigger in the sleep-wake cycle, with the most optimal concentration of CA 5 μg/mL. Exposure to CA extract was also able to reduce the expression of ERK (94795 ± 30830) and Akt (60113.5 ± 27833.5) with an optimum concentration of CA 2.5 μg/mL. Conclusion: Exposure to CA extract was able to improve the sleep activity of zebrafish larvae insomnia model by extending the total inactivity time (cumulative duration) and shortening the duration of first sleep (latency to first) in light and dark phases through inhibition of orexin, ERK, p38, and Akt.

AIM: To evaluate the effect of epigallocatechin gallate (EGCG) in preventing lens opacity and the aggregation of lens αB-crystallin in model rats of diabetes mellitus (DM). METHODS: This experimental study included Wistar rats for DM as in vivo models and divided into 5 groups. The treatment groups were administered EGCG by orally for 20d and were then assessed for their degree of lens opacity with binocular microscope and lens αB-crystallin expression from Western blot analyze. RESULTS: Pearson correlation test and regression analysis on EGCG exposure and final random blood sugar (RBS) obtained a significance level of P

The aim of this study was to investigate the effects of mobile phone electromagnetic radiation (MP-EMR) on the thyroid glands and hormones in Rattus norvegicus brain in term of thyroid function, reactive oxygen species (ROS), and monocarboxylate transporter 8 (MCT8) concentration. Forty rats were divided into different groups: control (without EMR exposure), EMR1 (120-min/day exposure), EMR2 (150-min), and EMR3 (180-min). The levels of serum thyroid stimulating hormone (TSH), thyroxine (T4), and malondialdehyde (MDA) and brain and MCT8 were measured using enzyme-linked immunosorbent assay. One-way analysis of variance followed by the Duncan test was used to analyze the data. Our data indicated that the levels of serum TSH and T4 in all the EMR groups were lower significant postexposure compared to the control with P < 0.01 (EMR1 and EMR2) and P < 0.001 (EMR3), suggesting hypothyroidism due to MP-EMR exposure. Increased MDA and decreased MCT8 levels were also observed following the intervention; however, the changes in both concentrations were notably significant after being subjected to 150-min and 180-min of exposure. In conclusion, a significant reduction in TSH, T4, and MCT8 levels indicated thyroid dysfunction due to MP-EMR exposure.

Background: In tobacco, nicotine is the primary toxic agent that causes health problems. As a primary metabolite of nicotine, cotinine is frequently used as an accurate measure of exposure to tobacco smoke. This study aims to find the factors affecting the serum cotinine level of male smokers in Malang, Indonesia. Methods: This cross-sectional study was conducted on 183 men who smoke regularly in Malang, Indonesia. They were selected by a purposive sampling method. Nicotine dependency and cotinine level were measured using the Fagerstrom test for nicotine dependence and the human cotinine ELISA Kit, respectively. Multiple linear regression analysis was performed to determine the effect of age, smoking duration, cigarette type, and the nicotine dependence on the continue level. ANOVA and independent t-test were also used for testing the study hypothesis. The significance level was set at 0.05. Results: Based on the ANOVA results, the factors of age, smoking duration, type of cigarette, and nicotine dependence together had a significant effect on the cotinine level (P

Background and Aim: The morbidity and mortality of Shigella infections remain a global challenge. Epitope-based vaccine development is an emerging strategy to prevent bacterial invasion. This study aimed to identify the ability of the 49.8 kDa pili subunit adhesin protein epitope of Shigella flexneri to induce an intestinal immune response in mice. Materials and Methods: Thirty adult male Balb/c mice were divided into a control group, cholera toxin B subunit (CTB) group, CTB+QSSTGTNSQSDLDS (pep_1) group, CTB+DTTITKAETKTVTKNQVVDTPVTTDAAK (pep_2) group, and CTB+ ATLGATLNRLDFNVNNK (pep_3). We performed immunization by orally administering 50 μg of antigen and 50 μl of adjuvant once a week over 4 weeks. We assessed the cellular immune response by quantifying T helper 2 (Th2) and Th17 using flow cytometry. In addition, we assessed the humoral immune response by quantifying interleukin (IL-4), IL-17, secretory immunoglobulin A (sIgA), and β-defensin using enzyme-linked immunoassay. Statistical analysis was performed using one-way analysis of variance and Kruskal–Wallis test. Results: Peptide oral immunization increases the cellular immune response as reflected by the increase of Th2 (p=0.019) and Th17 (p=0.004) cell counts, particularly in the CTB_pep_1 group. Humoral immune response activation was demonstrated by increased IL-4 levels, especially in the CTB+pep_3 group (p=0.000). The IL-17 level was increased significantly in the CTB+pep_1 group (p=0.042). The mucosal immune response was demonstrated by the sIgA levels increase in the CTB+pep_3 group (p=0.042) and the β-defensin protein levels (p=0.000). Conclusion: All selected peptides activated the cellular and humoral immune responses in the intestine of mice. Further studies are necessary to optimize antigen delivery and evaluate whether the neutralizing properties of these peptides allow them to prevent bacterial infection.

This study aimed to evaluate the effects of ciplukan (Physalis angulata L.) leaf ethanol extract on L-NG-nitroarginine methyl ester (L-NAME)-induced hypertensive rats. We randomly divided twenty-five Wistar rats into five groups. The sham group was given a PBS injection. The hypertensive group was injected with L-NAME on days 1 to 28. Three groups of hypertensive rats were given the extract on days 4 to 28. Blood pressure was measured using the tail-cuff method on days 0, 4, 10, and 27. The endothelial progenitor cells (EPCs) in the blood were measured by flow cytometry as a percentage of circulating angiogenic cells (CACs, CD34+/CD309+/CD45+) and endothelial colony-forming cells (ECFCs, CD34+/CD309+/CD45-). Serum NO and MDA levels, as well as serum SOD activity, were measured colorimetrically. Serum TNF-α levels were measured by the ELISA method. The ciplukan leaf extract reduced systolic and diastolic blood pressure, reduced the percentage of EPCs in the blood, increased serum NO levels, reduced MDA levels, increased serum SOD activity, and reduced serum TNF-α levels in L-NAME-induced hypertensive rats. It is concluded that ciplukan ethanol leaf extract exerts protective effects on L-NAME-induced hypertensive rats. These study results can strengthen the scientific basis of using ciplukan leaf ethanol extract to treat hypertension.

Context: Cataracts are currently the main cause of blindness worldwide. This condition caused by a low concentration of reduced glutathione (GSH) in the lens. Aims: To evaluate the effect of glutathione (GSH) on the hyperglycemia-induced oxidative stress, reticulum endoplasmic stress, and glycation in human lens epithelial cells. Methods: Human lens epithelial cells were cultured under high-glucose conditions. Malondialdehyde (MDA), protein carbonyl content (PCC), glucose reactive protein (GRP), and advanced glycation end product (AGE) were measured by enzyme-linked immunosorbent assay after 72 h of incubation for MDA, PCC, GRP and after 2 weeks incubation for AGE. Results: The MDA and PCC levels increased in response to high-dose glucose administration compared to the control group. MDA and PCC levels were decreased at all GSH doses, whereas the lowest mean MDA and PCC levels were observed at GSH doses of 10 and 100 μM, respectively. GRP levels increased after high-glucose administration as compared to the control group. Additionally, the groups co-treated with 30 and 100 μM GSH showed reduced PCC. The AGE level was reduced at all doses of GSH compared to those in the control group. Conclusions: The results suggest that GSH inhibits oxidative stress, reticulum endoplasmic stress, and AGE formation, which may lead to the progression of diabetic cataract. Additionally, GSH may maintain lens transparency by acting as an antiglycation and controlling the AGE formation.

Lead exposure (Pb) may aggravate the decrease in serum SOD levels, memory function reduction and social interaction in autism. This study aims to determine the effect of Moringa oleifera on autism models of rat exposed to lead. The five groups of rat exposed to tin were studied: normal rat (C -), autistic rat (C +) and autism rats given 180 mg (T1), 360 mg (T2) or 720 mg (T3) oleifera leaves powder. Lead acetate is given through a filler tube (0.5 gr / kg) and powdered moringa leaves mixed with feed. The results showed that serum SOD levels were lower in autistic rat than normal rat and intake of Moringa olifeifera leaves powder increased serum SOD levels in autistic rats but was not seen in normal rat. The autistic rat had higher lead levels but lower zinc levels, which were not influenced by Moringa olifeira. The social interaction between rats is strongly influenced by unbidden lead with Moringa olifeira, both in normal rat and in autistic rat. Moringa oleifera will increase serum SOD levels in autistic rats but not at normal levels. Moringa olifeira does not protect significantly rat from lead causing less social interaction, nor does it lower serum Pb levels or increase Zn levels. Moringa leaves powder does not affect memory significantly.

AIM:To evaluate the effects of reduced glutathione(GSH)and niacin combination on protein oxidative stress, endoplasmic reticulum(ER)stress, glycation, and aggregation of the αβ crystalline in human lens epithelial(HLE)cells treated with high glucose levels. METHODS:HLE cells were cultured and exposed to 25 mmol/L glucose to promote high glucose conditions. Groups of cells were co-treated with three different combinations of dosages: 10 μmol/L GSH+2 μmol/L niacin, 30 μmol/L GSH+25 μmol/L niacin, and 100 μmol/L GSH+25 μmol/L niacin. After 72h incubation, protein carbonyl content(PCC)and glucose reactive protein(GRP78)content were assessed using ELISA examinations. After two-week incubation, advanced glycation end products(AGEs)were also assessed and the expression of αβ crystalline was measured using Western Blot examination. The SPSS 18.0 statistical package was used for all data analyses.RESULTS:PCC and GRP78 levels in the co-treated groups were not significantly reduced compared to control(P>0.05). In contrast, there was a significant decrease of the AGEs levels in all groups co-treated with GSH and niacin when compared with the control group(PCONCLUSION:The results suggest that combinations of GSH and niacin inhibit the aggregation of proteins and prevent glycation in hyperglycemic human lens epithelial cells. This study shows that this combination may play an active role in preventing diabetic cataract mainly from the AGEs pathway.

Citicoline, presumed to be involved in the dopaminergic pathway, might play a role as a candidate agent in controlling myopia. However, its study with respect to myopia is limited. The aim of this study is to demonstrate the effect of citicoline on the expression of MMP-2, TGF-β1, and Ki-67, and on the thickness of scleral tissue of a rat myopia model. Immunohistochemistry was performed to evaluate the expression of MMP-2, TGF-β1, and Ki-67 as the markers for fibroblast proliferation. Hematoxylin and eosin staining were used to evaluate scleral thickness. An electronic digital caliper was used to evaluate the axial length. The treatment group administered with 200 mg/kg BW/day had the lowest mean MMP-2 expression, axial elongation, and fibroblast proliferation, but it had the highest mean scleral thickness. The treatment group administered with 300 mg/kg BW/day had the highest mean TGF-β1 expression. Citicoline is able to decrease MMP-2 expression and fibroblast proliferation and increase TGF-β1 expression and scleral tissue thickness significantly in the scleral tissue of rat models for myopia.

AIM: To investigate the effect of Physalis angulata leaf extract on apoptotic and proliferation of retinoblastoma cells. Despite several previous studies evidencing the anti-cancer potential of Physalis angulata; however, certain study that proves its benefits in retinoblastoma cancer cells has been limited. METHODS: This study utilizes an in-vitro experimental study by applying Y79 human retinoblastoma cell line culture obtained from the American Type Culture Collection (ATCC; 10801 University Boulevard Manassas, VA 20110, USA). The cell was divided into 4 groups. Group I was the control group without the administration of Physalis angulata leaf extract. Whereas, group II, II and IV are engaged with 25, 50, and 100 µg/mL of Physalis angulata leaf extract respectively. After a 24h incubation, an examination with microtetrazolium (MTT) cell proliferation assay and Annexin V apoptosis detection was conducted. Statistical analysis was performed with the Tukey test. RESULTS: Physalis angulata leaf extract improved apoptosis and significantly reduced the number of living cells in retinoblastoma cells, along with the increase in the given dose. Based on the Tukey test, a significant difference was found in the treatment group at 50 µg/mL (P=0.025) and 100 µg/mL (P=0.001) in the measurement of apoptosis. Proliferation measurements also indicated a significant decrease in the number of living cells in the 50µg/mL treatment group (P=0.004), and in the 100 µg/mL treatment group (P=0.000). Meanwhile, a dose of 25 µg/mL indicated insignificant difference in the two measurements. Improved apoptosis and decreased number of living cells occured at a dose of 100 µg/mL. Decreased number of living cells (in the measurement of proliferation) was due to the inhibited proliferation or improved apoptosis. CONCLUSION: Physalis angulata leaf extract improve apoptosis in retinoblastoma cell culture, requiring further research to inhibit proliferation.

Diabetes is a disorder in the metabolism of carbohydrates, lipids and proteins with various causes. From the previous research, GPR43 is a potent clinical target for the treatment of type 2 diabetes. It can be activated by Short Chain Fatty Acid (SCFA) which is an organic fatty acid produced through fermentation of dietary fiber by bacteria in the distal intestine. The aim of this study was to analyze the effect of C680T GPR43 Gene Variations to Its Interaction with Short Chain Fatty Acid (SCFA) in silico. The study used 8 sequences of GPR43: ID AF024690, AC002511, EU432114, BC095535, BC096198, BC096199, BC096200, BC096201 and data of ligand (SCFA): acetic acid (CID 176), Butyric acid (CID 264) and Propionic acid (CID 1032). GPR43 was modelled using I-TASSER, Sequence and structural alignments were conducted using Bioedit and Superpose V.10, respectively. The Docking process was done using PyRx and molecular interaction was analyzed using Discovery Studio 2016. The Result showed that three types of SCFA bind to GPR43 variants T and GPR43 variant M obtained a similar pattern. The binding affinity from the largest to the smallest is butyric acid > propionic acid > acetic acid. GPR43 variant T has greater affinity to the SCFA than the GPR43 variant M. There is no differences of preference between GPR43 variant T and GPR43 variant M to bind SCFA.

The thyroid-stimulating hormone receptor (TSHR) is one of the thyroid antigens responsible for Graves’ disease and acts as a biomarker for early detection. The purpose of this study was to computationally compare the effectiveness of TSHR56 and TSHR169 fragments in binding to the thyroid-stimulating human monoclonal autoantibody (M22) at the molecular level. The 3D model of M22 was obtained from RCSB (ID 3G04), while the TSHR antigen models were submitted to homology modeling using SWISSMODEL and PhyreV2.0 server to predict the protein structures. The model was validated by generating a Ramachandran plot with the RAMPAGE server. Prediction of the molecular interaction between TSHR and M22 was performed using the HADDOCK web server (version 2.2). Analysis of the binding affinity was conducted using the PRODIGY server. Interactions within the TSHR-M22 complex were analyzed using DIMPLOT. The antigen models had reliability scores of 100% and 97.7% for TSHR56 and TSHR169, respectively. The results of the molecular docking analysis revealed a better HADDOCK score for TSHR169 (-144.7 ± 2.4) compared to TSHR56 (-53.3 ± 10.1). However, the affinity of TSHR56 (-10.1 Kcal/mol) and TSHR169 (-10.2 Kcal/mol) were not significantly different. The results of the TSHR-M22 interaction specificity analysis suggested that TSHR169 is superior to TSHR56, as the number of interacting amino acids was comparable to the control (TSHR260-M22). It can be concluded that TSHR169 represents a specific full-length TSHR antigen and can be developed as biomarker for Graves’ disease.

Background: Serum Glial Fibrillary Acidic Protein (GFAP) is a great potential for biomarker that is widely studied as a diagnostic biomarker of acute stroke. Sampling within 6 hours after onset is the best time window, but in Indonesia, stroke patients often arrive late more than 6 hours. Objective: To identify the difference in time of blood sampling with serum GFAP levels within 24 hours onset of ischemic stroke (IS) patients and intracerebral hemorrhage (ICH) strokes. Methods: Cross-sectional analysis with purposive sampling, sampling in IS and ICH strokes that arrive at the ER within 24-hour on-site. The serum GFAP examination was performed with ELISA. Results: In this study, 41 acute stroke patients with 24-hour onset of each stroke were grouped into group 1 (

Objective: To analyze the effects of subchronic Cypermethrin on the ovary and endometrium as well as the involvement of apoptosis in the toxicity of cypermethrin. Methods: A total of 32 female Wistar rats were randomly divided into four groups, with 8 rats in each group. The control group received no treatment, and the other three groups received oral cypermethrin at 10, 15 or 20 mg/kg body weight for 28 days (sub-chronic). The granulosa cells were calculated histopathologically. The apoptotic index was determined by in situ technique. Histopathological examination was performed on the uterus and ovary. Results: There was no significant difference in the number of primary follicular granulosa cells between the treatment groups and the control group (P>0.05). However, the number of secondary and tertiary follicle granulosa cells in the treatment groups was significantly decreased compared to that of the control group (P all

Abstract: Rhodamin B is harmful to the body because it is proven can increase the oxidative stress on various reproductive chain. Rhodamin B contains compounds of chlorine (Cl), is dangerous and halogen compounds are reactive. Damage to the hypothalamus will interfere with the secretion of FSH and LH cause the follicle ripening so distracted that result in a decrease in the hormone estrogen. The hormone estrogen plays an important role in the menstrual cycle and proliferation of the endometrium. This research aimed to prove the effect of Extra Virgin Olive Oil (EVOO) with the expression of estrogen receptors α and endometrial thickness of Rattus norvegicus exposed by rhodamin B. The design used true experimental with an approach of post test only control group design. The research used white rat (Rattus norvegicus wistar strain) as the sample. The results showed that EVOO could become an alternative to prevent oxidative stress due to exposure of rhodamin B. Keywords: Rhodamin B, Extra Virgin Olive Oil (EVOO), Estrogen receptors α, the thickness of the Endometrium

Abstract: The use of dyes in food today is very apprehensive. Many foods contain dyes that are prohibited to be used in food. The addition of the dye will have an impact on health. The aim of this research was to know the effectiveness of the administration of extra virgin olive oil to the number of granulosa cell and 17β estradiol level of Rats exposed by rhodamin B. The design of this study was true experimental with post test approach of group design control. The subject of the study was 30 white mice Rattus novergicus in the age of 12 weeks weight average 220 grams and in healthy condition. Rats were divided into 5 groups. Control group without treatment, positive control treatment with rhodamine B 18 mg / 200gr. Treatment 1 administration of rhodamine B 18 mg / 200gr and EVOO (Extra Virgin Olive Oil) 1.5 ml / KgBB, treatment 2 rhodamine B 18 mg / 200gr and EVOO 3 ml / KgBB, treatment 3 rhodamine B 18 mg / 200gr and EVOO 4,5 ml / KgBB. The number of granulosa cells was determined by HE staining, the level of 17β estradiol with ELISA. The data was analyzed by Kruskal Wallis test, Spearman Rank and Simple Linear Regression with degree of significance α

Background — Cigarette smoking negatively impacts the health of both active smokers and passive smokers. ROS (Reactive Oxygen Species) from cigarette smoke can cause damage to the ovary and inhibit ovarian follicle growth. Objectives — This study aimed to assess the effects of the anthocyanins from the sweet potato (Ipomoea batatas L.) purple cultivar on ovarian malondialdehyde (MDA) and 17β-estradiol levels in rats (Rattus norvegicus) exposed to cigarette smoke. Material and Methods — The Rattus norvegicus Wistar strain was used in this study. Thirty rats were divided into five groups. Cigarette smoke was given as two cigarettes/day for four minutes, in the morning and afternoon for eight weeks after rats were at the proestrus stage. MDA levelwere measured with the BIOXYTECH MDA-586TM spectrophotometric assay for malondialdehyde, whilst 17β-estradiol was measured by ELISA. Data were then analyzed using one-way ANOVA. Results — MDA level were significantly different between the negative control and positive control groups based on double comparison with the LSD test (p=0.000

Objective: This study aimed to determine the effect of Extra Virgin Olive Oil (EVOO) on vasodilator enzyme by repairing angiogenic function in rat model of preeclampsia. Materials and methods: This research consisted of five groups; negative control (normal pregnant rats) group, positive control (preeclampsia rat model) group, preeclampsia rat model groups given EVOO in 3 different doses (0.5 ml/day, 1 ml/day, and 2 ml/day, respectively). Blood pressure measurements were carried out on day 12, 15, and 19 of pregnancy. After the rats were sacrificed, the placentas were collected to determine endothelial Nitric Oxide Synthase (eNOS) level of maternal plasma to determine soluble Fms-like Tyrosine Kinase 1 (sFlt-1) and Vascular Endothelial Growth Factor (VEGF) level. Results: There were significant higher sFlt-1 level (p < 0.001), lower VEGF level (p = 0.009), and lower eNOS level (p = 0.034) between negative and positive control groups. After EVOO administration, sFlt-1 level was lower in dose 1 and 2 groups but higher in dose 3 group in accordance with VEGF and eNOS levels that were increasing both in dose 1 and dose 2 groups but decreasing in dose 3. There were significant differences between positive control and dose 1 (p = 0.015) and dose 2 (p = 0.001) in sFlt-1 level. None of all dose groups were statistically different with positive control group in VEGF level (dose 1 p = 0.601; dose 2 p = 0.297; dose 3 p = 0.805). eNOS levels of all dose groups were statistically different from that of the positive control group (dose 1 p = 0.014; dose 2 p = 0.001; dose 3 p = 0.024). Conclusion: Administration of EVOO modulates eNOS as vasodilator enzyme by repairing the angiogenic function indicated by decreased sFlt-1 level and increased VEGF in rat model of preeclampsia.

The purpose of this study was to investigate the effect of genistein administration on the modulation of the estrogen receptor, inhibition of inflammation and angiogenesis in the murine model of peritoneal endometriosis. A total of thirty-six mice (Mus musculus) were divided into six groups (n = 6), including the control group, endometriosis group, endometriosis group treated with various doses of genistein (0.78; 1.04; 1.3 mg/day), and endometriosis group treated with leuprolide acetate (0.00975 mg/day every 5 days for 15 days). Analysis of estrogen receptor-α, estrogen receptor-β, TNF-α, IL-6, VEGF, and HIF-1α were performed immunohistochemically. Expression of estrogen receptor-α, estrogen receptor-β, TNF-α, IL-6, VEGF and HIF-1α increased significantly compared with the control group (p 0.05). Genistein also decreased the expression of TNF-α and IL-6 (1.04 and 1.3 mg/day) compared with the endometriosis group, reaching level comparable to that of the control group (p > 0.05). It was concluded that genistein is able to modulate estrogen receptor-α and estrogen receptor-β and inhibit the development of inflammation and angiogenesis in the murine model of peritoneal endometriosis. Thus, genistein can be a candidate in the treatment of endometriosis. Keywords: Estrogen receptor, Growth factor, Inflammation, Angiogenesis, Peritoneum

Background. One of pediatric emergencies that has high mortality is status epilepticus (SE). Correlation between tumor necrosis factor (TNF)-α, interleukin (IL)-6, and SE had been reported but human study of it is limited. Objective. To compare TNF-α and IL-6 level in children with SE to those of children without SE and to find correlation between both cytokines. Methods. Cross-sectional study was conducted in dr.Saiful Anwar Hospital Malang with 48 children were enrolled in this study. All subjects were divided into three groups, including children who had SE; children who had seizure but not SE; and children who have no seizure. The levels of TNF-α and IL-6 serum were measured by ELISA. Results. TNF-α and IL-6 serum level were not significantly different between groups (p=0.920, p=0.829). We found interesting fact that the level of IL-6 in children with SE who have no disability was significantly higher than that of children who died or had disability (p=0.015). There was strong correlation between TNF-α and IL-6 in SE group (R 2 = 0.841 and p = 0.0001). Conclusion. IL-6 serum level was higher in SE children who have no disability and correlate with TNF-α serum level.

AIM: To investigate the mechanism of pericyte migration through Angiopoietin-2 (Ang-2)/Tie-2 signaling pathway. METHODS: We divided the rats into 5 groups. Each diabetic rat model groups injected with Tie-2 inhibitor, ERK1/2 inhibitor, Akt/PKB inhibitor, and DMSO intravitreal. Retinal digest preparation was done to examine the retinal vasculature including pericyte: endothelial ratio, and morphology of pericyte migration. Tie-2, ERK1/2 and Akt/PKB phosporylation were analyzed by confocal laser scanning microscopy. RESULTS: There was a correlation between pericyte migration with increasing Ang-2 (P

Background. Seizures are a common clinical manifestation in the emergency room. Nearly 5% of children aged under 16 years. Seizures are important as a sign of neurological disorders. Members of interferon are widely studied with seizures and epileptogenesis is interferon-γ. In a study by Choi in 2011, a study of seizures in humans showed that the condition of status epilepticus increased levels of interferon-γ which is quite high compared to patients not status epilepticus seizures. In response to the aftermath of the seizure, Interferon-γ system induces the formation of IL-10 acts as an anti-inflammatory agent that aims to put an end to the action of Interferon-γ. Objective. To know role of Interferon-γ and IL-10 in children with status epilepticus. Methods. This research using cross sectional design with recruiting 30 study sample consisted of 15 children in the seizure group SE and 15 children in the group without SE seizures. Measured levels of IFN-γ and interleukin-10 by ELISA. Results were analyzed with the Mann-Whitney and corelation Spearman to see the relationship levels of IFN-γ and IL-10 in children with status epilepticus with SPSS-23. Results. The results showed significant differences between groups SE and convulsive seizures non SE ;IL10 (p= 0.000) and IFN-γ (p=000). Conclusion. There are significant correlation between the levels of IFN-γ and IL-10 with status epilepticus.

Currently, umbilical cord mesenchymal stem cells have the potential to be used as treatment options for any cartilage lesion. This research aimed to evaluate the effects of conditioned medium from umbilical cord mesenchymal stem cells (UC-MSC) on damaged cartilage through the expression of proteins TGF-β1, SOX-9, type II collagen and aggrecan, which are known to be related to chondrogenesis. UC-MSC were isolated from 19-days-pregnant Wistar mice and were cultured using the standard procedure to obtain 80% confluence. Subsequently, the culture was confirmed through a microscopic examination that was driven to be an embryoid body to obtain a pre-condition medium. This research utilized 3-month-old male Wistar mice and was categorized into 6 groups (3 control and 3 treatment groups). Each animal had surgery performed to create a femur condyle cartilage defect. The treatment groups were administered a dose of stem cells at 1 mL/kg. Next, immunohistochemical (IHC) staining was performed to examine the expression of TGF-β1, SOX-9, type II collagen and aggrecan in the 2nd, 3rd, and 4th month of evaluation. The results were analyzed statistically using ANOVA test. For each of the treatment groups, there was increased expression (p < 0.05) in all proteins TGF-β1, SOX-9, type II collagen and aggrecan when compared with control groups at the 2nd, 3rd, and 4th month of evaluation. Pre-conditioned medium from UC-MSC potentially increases the expression of TGF-β1, SOX-9, type II collagen and aggrecan in the damaged cartilage of Wistar mice.

Proinflammatory cytokines are elevated in status epilepticus and febrile seizure and associated with tissue damage. This study aimed to investigate the differences in interleukin 1 beta (IL-1β) and interleukin 1 receptor agonist (IL-1ra) levels in status epilepticus as compared with febrile seizure and febrile. This cross-sectional study was designed to include 45 subjects divided equally into three groups (status epilepticus, febrile seizure, and febrile). Both IL-1β and IL-1ra were measured by using an ELISA method. Results showed that IL-1β levels were significantly higher in the status epilepticus group as compared with the febrile seizure and febrile groups (p < 0.05). IL-1ra levels in the status epilepticus group were significantly lower compared with the febrile seizure group (p = 0.04). Consistently, the IL-1β/IL-1ra ratio in the status epilepticus group was significantly higher as compared with the febrile seizure group (p = 0.01). We concluded that IL-1β and the IL-1β/IL-1ra ratio were significantly higher in status epilepticus. IL-1ra levels were significantly higher in the febrile seizure group.

In primary open angle glaucoma, decrease of hyaluronic acid level is related to extracellular matrix deposition of trabecular meshwork. Extracellular matrix deposition will increase humor aqueous outflow resistance which in turn increase the intraocular pressure. MMP-9 as one of matrix metalloproteinase (MMP) family, has a high potency to degrade the extracellular matrix. The expression of MMP-9 can be activated by hyaluronic acid through its binding with CD44 as hyaluronic main receptor. The aim of this research is to determine the effect of hyaluronic acid on MMP-9 expression in trabecular meshwork cell culture of patient with primary open angle glaucoma by immunocytochemical method. Trabecular meshwork cell culture were exposed to hyaluronic acid at four different concentration (0mg/ml, 1 mg/mL, 3 mg/mL, 6 mg/mL). After 24 hours incubated, we find a significant difference of MMP-9 expression between control group and group with hyaluronic acid. There was significant correlation between hyaluronic acid and MMP-9 expression. Hyaluronic acid exposure will increase MMP-9 expression in trabecular meshwork cell culture. This finding suggest that hyaluronic acid can influence the resistance of humor aqueous outflow in trabecular meshwork by up-regulating MMP-9 expression, and therefore will increase extracellular matrix degradation. Key words : hyaluronic acid, primary open angle glaucoma, MMP-9

Background. Tuberculosis caused by mycobacterium tuberculosis. The support of these inflamated factors contained in this focus determine the prognosis of TB towards Central Nervous System. One of the inflammation factors is TNF-α. Objective. To find the expression of TNF-α on the brain which are infected by M. tuberculosis. Methods. Experimental study, Wild Mus musculus to compare the group which has got infected before. The similiar study has been conducted by Laksmi Wulandari, pulmonologist. The observation towards the expression of TNF-a into a mouse’s brain was done by using the imunohistokimia method. It is marked by the brown color in the core, sitoplasma and the cell with upgrading size 100x times 20. Results. This is evident that significanly corelation (P = 0.000) between independent (Mycobacterium tuberculosis infection at incubation periode 8 and 16 weeks) and dependent variabel (TNF-α expression on brain winstar tissue that infected by Mycobacterium tuberculosis for 8 and 16 weeks). Conclusion. Strong relationship between expression of TNF-a and the M. Tuberculosis’ infection.

Cervical cancer recurs in 90% cases and linked to cancer stem cells that able to self-renew and responsible for recurrence, metastasis, and mortality of cancer. Isolation and identification of cancer stem cells using serum-free medium needs expensive growth factors and consume time. This study try to grow tumor sphere using culture plate coated with 1% agarose as an efficient and economical alternative to non-adhesive culture plate. HeLa cell line was grew in culture plate coated with 1% agarose and non-adhesive culture plate using similar medium and culture condition. Tumor spheres morphology was observed and the colonies were counted in 7 days followed by single cell assay. Tumor spheres then counted for CD133+, CD34+, and Sox2 expression using flowcytometry. Culture plate coated with 1% agarose can be used as an economic and efficient alternative to culture tumor sphere. Using culture plate coated with 1% agarose, the tumor spheres formed in 7 days with similar morphology to non-adhesive culture plate. Tumorsphere had three dimensional – sphere shape that tightly attached, colonized, and overlapped. The tumor sphere colony counts of two plates were statistically have no significant difference (p=0,667). Single cell assay of a tumor sphere shows that it can grow new tumor spheres with similar morphology. The tumor sphere from culture plate coated with 1% agarose express CD133+ and CD34+ as much as 8.78% ± 2.14 and Sox2 as much as 35.30% ± 23.82 whereas tumor sphere from non-adhesive culture plate express CD133+ and CD34+ as much as 62.36% ± 1.06 and Sox2 as much as 98.86% ± 0.56 (p = 0000).

AIM: To evaluate effect of hypertension on retinal ganglion cell (RGC) apoptosis, intraocular pressure (IOP), and the activation of endothelin-1 (ET-1) signaling pathway in central retinal artery (CRA) in rats. METHODS: The experimental study was performed on 20 male Sprague Dawley rats that were divided into control group, and hypertension groups. The hypertension was induced by subcutaneous deoxycorticoacetate (DOCA) 10 mg/kg twice a week and administered 0.9% NaCl solution daily for 2, 6, and 10wk. Blood pressure (BP) was measured using animal BP analyzer. IOP was measured by handheld tonometry. Retinal tissue preparations by paraffin blocks were made after enucleation. The expression of ET-1, eNOS, ET-1 receptor A (ETRA), ET-1 receptor B (ETRB), and phosphorylated myosin light chain kinase (MLCK), and caldesmon (CaD) in CRA and RGC apoptosis were evaluated through immunofluorescent staining method then observed using laser scanning confocal microscopy. RESULTS: BP significantly increased in all of the hypertension groups compared to control (P=0.001). Peak IOP elevation (7.78±4.14 mm Hg) and RGC apoptosis (576.15±33.28 Au) occurred on 2wk of hypertension. ET-1 expression (1238.6±55.1 Au) and eNOS expression (2814.2±70.7 Au) were found highest in 2wk of hypertension, although the ratio of ET-1/eNOS decreased since 2wk. ETRA reached peak expression in 10wk of hypertension (1219.4±6.3 Au), while ETRB significantly increased only in 2 weeks group (1069.2±9.6 Au). The highest MLCK expression (1190.09±58.32 Au), CaD (1670.28±18.36 Au) were also found in 2wk of hypertension. CONCLUSION: Hypertension effects to activation of ET-1 signaling pathway significantly in CRA, elevation of IOP, and RGC apoptosis. The highest value was achieved at 2wk, which is the development phase of hypertension.

There is considerable evidence, suggest that, consumption of food additives monosodium glutamate (MSG), a flavor enhancer was unhealthy. Herbal medicine Nigella sativa (NS) has antioxidant properties able to cure the toxic induced by MSG. This study aimed to evaluate the risks of excessive use of MSG and to study the role of NS to inhibit inflammation and renal damage. Treated rats (twenty four male wistar rats) were divided into six group and analyzed by measuring the cells in blood, interleukin-10, interferon-γ serum levels by ELISA method and remove kidneys for histological examination. Histological of kidney for all groups except control, were showed different abnormalities include congestion of some blood vessels, hemorrhage between tubules, widening in the renal tubules, revealed severe dilatation of Bowman's capsule and shrinkage of glomeruli, and areas of huge vacuole, were observed compared with control. Interleukin-10 was reduced in Groups 2,3,4 and 5, whereas increase in NS group compared with control. Interferon-γ was increased in groups 2,3,4 and reduced in groups 5,6 compared with control. Eosinophil was increased in groups 2,5 and reduced in groups 3,4, 6 compared with control. This present study showed that administration of MSG to rats induced many changes effects on inflammatory cells, cytokines and histological of kidneys. NS has benefit in blood parameters, whereas harmful on kidney at these doses.

Monosodium glutamate (MSG) is a commonly used food additive and found in most soups, fish, and processed meat. The use of MSG in food is growing. However, the fear of consuming MSG has increased in the last few years due to the adverse reactions and toxicity in the liver. Nigella sativa (NS) is used as traditional medicine for the treatment of many diseases. It has been extensively investigated in recent years due to its notable pharmacological properties such as inhibit oxidative stress. The present study was undertaken to investigate the effect of different doses of Nigella Sativa on alpha KGDH activity and liver histology of MSG-induced rats. The animals (n=30) were grouped into A (control), B (treated with MSG 1g/kg.bw ), C (treated with MSG 1g/kg.bw and NS 0.1 g/kg.bw), D (treated with MSG 1g/kg.bw and NS 0.2 g/kg.bw), E (treated with MSG 1g/kg.bw and NS 0.4 g/kg.bw) and F (given a daily NS extract 0.2 g/kg.bw). Alpha KGDH activity was investigated using ELISA method and liver histopathology by light microscope. The MSG treatment increased Alpha KGDH activity and disturbed liver architecture, hemorrhage in the central veins, areas of necrosis, vacuolation and increased inflammatory cells infiltration. The condition was normalized by treatment NS on dose 0.2 and 0.4 g/kg.bw. The findings showed that the administration of MSG increases alpha KGDH and induces damage in liver tissue. Nigella sativa extract can reduce alpha KGDH and prevent liver damage caused by MSG.

Purpose: The aim of this study were to determine the association of increasing of FasL with increasing proliferation of retinoblastoma cells. Materials and Methods: The protein expression was analyzed in 30 retinoblastoma samples from paraffin block using immunohistochemical method for evaluation of FasL, CDK4, and Ki-67 expression. Results: Among 30 retinoblastoma samples, FasL expression majority was negative in 33.3 % (10 samples) and strong in 36,8 % (11 samples). CDK4 majority 53,3% was weak expression and Ki-67 was high expression also in 53,3% (16 samples). The expression of FasL was significantly related to CDK4 (r: 0.363; p: 0.048). The CDK4 was also significantly related to Ki-67 expression (r: 0.601; p: 0.000). Conclusion: The increasing of FasL on the mechanism Fas counterattack induces proliferation of retinoblastoma cells. [Cukurova Med J 2014; 39(4.000): 788-794]

Cervical cancer is one kind of many cancers that cause death to women around the world. Many studies had support the statement that inflammation has a strong linkage with cancer development. Several factors like proinflammatory factor can influence tumor cell microenvironment, and induce a faster proliferation. TNF-α is suspected can induce proliferation. While cancer itself can induce inflammation, which is marked by several marker. One of them is HMGB1, released from the cell as active secretory lysosomes or passive diffusion. Genistein has demonstrated growth inhibitory effects of various types of cancer cells. It inhibits tyrosine kinase pathway, which can be activated by TNF-α. One of those pathways that have the link with proliferation is p38. This study tries to reveal about inhibitory effect of genistein toward p38 pathway that had been activated by TNF-α. This research was conducted by exposing cultured HeLa cells with various doses of genistein for 90 minutes, and then exposed to TNF-α 10 ng / mL for 20 minutes. Observations were made with a confocal microscope, by staining the cells with pp38-TRITC and HMGB1 antibody. The intensity was measured and analyzed by Fluoview software. The results suggest that there be significant differences between pp38 intranuclear intensity and HMGB1 extranuclear intensity of each dose of genistein (p = 0.000, ANOVA). pp38 and HMGB1 intensity were increased along with increasing genistein dose, but at high dose there were noted decreasing of pp38 and HMGB1 intensity. At apoptotic dose, pp38 and HMGB1 intensity were increased markedly, showing the effect of apoptosis. In general, increasing doses of genistein increase intranuclear p38 activation and HMGB1 extranuclear translocation. So there were a strong linkage between p38 activation and HMGB1 translocation in this study.

Renin catalyzes the cleavage of angiotensinogen into angiotensin I. Genetic variant C-5312T of renin enhancer has been reported to increase in vitro renin gene transcription. However, no obvious in vivo study was performed to see the renin level in C-5312T when treated with angiotensin receptor blockers (ARB). Therefore, this study aimed to investigate the serum renin level and blood pressure response in ARB treated hypertensive patients. Single nucleotide polymorphism (SNP) of C-5312T was identified in 55 hypertensive patients by using multiplex PCR and renin serum level was assayed by ELISA. The data showed that the increase of serum renin levels after 5 months of ARB treatment was significantly higher in patients with CT/TT genotype (10 pg/mL) than those with CC genotype (4.08 pg/mL) (P = 0.025). Hypertensive patients with CT/TT genotypes also showed less diastolic pressure reduction than CC genotypes in hypertensive patients with valsartan treatment (P = 0.04) or telmisartan treatment (P = 0.03). Finally, these findings suggested that SNP of C-5312T REN enhancer might contribute to higher increased renin serum levels and less diastolic blood pressure response to ARB treatment.

Background: Insomnia is difficulty initiating or maintaining sleep for at least three nights a week or more and lasting for at least 3 months. One of the molecules that play a role in the circadian rhythm of arousal system is hypocretin/orexin. Orexin activates the p38-MAPK signaling pathway and increases phosphorylated ERK1/2 levels. Centella asiatica (CA) has a role in the signal work of the MAPK/ERK, Akt, and p38 path in many various diseases. Methods: The research method used is true laboratory experimental. The research approach used was randomized control group post-test only. Zebrafish embryos aged 0-7 dpf were used in this study. The treatment group consisted of 5 groups: normal, insomnia, insomnia + 2.5 μg/mL CA, insomnia + 5 μg/mL CA, and insomnia + 10 μg/mL CA. The locomotor motion of zebrafish larvae was observed using Basler cameras on days five-, six- and seven-day post fertilization (dpf), then analyzed by using Western Blot method. Results: The results proved that exposure to CA extract was able to reduce the expression of orexin (91963 ± 9129) and p38 (117425 ± 6398) as an arousal trigger in the sleep-wake cycle, with the most optimal concentration of CA 5 μg/mL. Exposure to CA extract was also able to reduce the expression of ERK (94795 ± 30830) and Akt (60113.5 ± 27833.5) with an optimum concentration of CA 2.5 μg/mL. Conclusion: Exposure to CA extract was able to improve the sleep activity of zebrafish larvae insomnia model by extending the total inactivity time ( cumulative duration) and shortening the duration of first sleep ( latency to first) in light and dark phases through inhibition of orexin, ERK, p38, and Akt.

Amaç: Bu çalışmanın amacı artan FasL ekspressyonu ile retinoblastom hücrelerindeki proliferasyon artışı arasındaki ilişkiyi incelemektir. Materyal ve Metod: Ki-67, CDK4 ve FasL ekspresyon değerlerini incelemek için parafin blok yapılmış 30 retinoblastom örneğinin protein ekspresyon değerleri immünohistokimyasal analiz ile belirlendi. Bulgular: 30 retinoblastom örneği arasından %33.3 ünde (10 örnek) FasL ekspresyonu yok iken %36,8 inde güçlü bir ekspresyon vardır. %53.3 (16 örnek) örnekte CDK4 ekspresyonunun zayıf olduğu görülürken Ki-67 ekspresyonunun yüksek olduğu tespit edilmiştir. FasL ekspresyonu yüksek oranda CDK4 ekspresyonu ile ilgilidir (r:0,363, p:0,048). Aynı zamanda CDK4 ün ekspresyonu da Ki-67 ekspresyonuyla ilgilidir (r:0,601, p: 0,000). Sonuç: FasL atağı mekanizmasındaki artan FasL ekspresyonu retinoblastoma hücrelerinin proliferasyonunu indüklemektedir

Background: Insomnia is difficulty initiating or maintaining sleep for at least three nights a week or more and lasting for at least 3 months. One of the molecules that play a role in the circadian rhythm of arousal system is hypocretin/orexin. Orexin activates the p38-MAPK signaling pathway and increases phosphorylated ERK1/2 levels. Centella asiatica (CA) has a role in the signal work of the MAPK/ERK, Akt, and p38 path in many various diseases. Methods: The research method used is true laboratory experimental. The research approach used was randomized control group post-test only. Zebrafish embryos aged 0-7 dpf were used in this study. The treatment group consisted of 5 groups: normal, insomnia, insomnia + 2.5 μg/mL CA, insomnia + 5 μg/mL CA, and insomnia + 10 μg/mL CA. The locomotor motion of zebrafish larvae was observed using Basler cameras on days five-, six- and seven-day post fertilization (dpf), then analyzed by using Western Blot method. Results: The results proved that exposure to CA extract was able to reduce the expression of orexin (91963 ± 9129) and p38 (117425 ± 6398) as an arousal trigger in the sleep-wake cycle, with the most optimal concentration of CA 5 μg/mL. Exposure to CA extract was also able to reduce the expression of ERK (94795 ± 30830) and Akt (60113.5 ± 27833.5) with an optimum concentration of CA 2.5 μg/mL. Conclusion: Exposure to CA extract was able to improve the sleep activity of zebrafish larvae insomnia model by extending the total inactivity time ( cumulative duration) and shortening the duration of first sleep ( latency to first) in light and dark phases through inhibition of orexin, ERK, p38, and Akt.

Background: This review aimed to emphasize the effects, mechanisms involved, conditions required to trigger impacts, and the adapted responses of EMF on thyroid gland. Methods: The included literatures were collected from the Oceania Radiofrequency Scientific Advisory Association (ORSAA) database which were published before April 2021. The searched was then refined with keyword ‘thyroid’. Results: A total of 15 experimental studies, 10 observational studies, 4 review articles, and 1 medical hypothesis were included. Hypothyroidism is the most reported functional abnormality. The identified morphological abnormalities of the thyroid gland include increased follicular epithelial and interfollicular tissue, decreased colloid volume of cubic cells, follicular fluid, interfollicular space, changes in lysosomes, granular endoplasmic reticulum, cell nuclei, changes in glandular structures including cell hypotrophy, glandular hypertrophy, and increased apoptosis via caspase-dependent pathways. The observational studies reported an increase in thyroid cancer incidence and the diameter of the left anteroposterior of the thyroid gland. Postulation that thyroid cancer is associated with thyroid cancer EMF exposure, is suggested by a medical hypothesis paper. The possibilities of hypothyroidism, thyroid cancer, and cell apoptosis due to EMF exposure have been highlighted in the review articles. Conclusion: Despite these findings, critical elements, such as the effects and mechanisms of EMF exposure on thyroid hormone transporters, genomic and non-genomic actions, conditions required for the effect to occur, and the resultant adapted responses, have not been explored suggesting the need of further research.

Background: The most common type of lips and oral cavity cancer is oral squamous cell carcinoma (OSCC). Doxorubicin (DX) is commonly used as a chemotherapy drug, but its use is limited due to risk factors and drug resistance. Chrysanthemum cinerariifolium (Trev.) (CC) has potential as an anticancer agent. Combining the plant extract and chemotherapy drug might prevent OSCC proliferation by inhibiting PI3K and cyclin D protein. Therefore, the present study aimed to determine the synergistic effect of the combination of C. cinerariifolium (Trev.) and doxorubicin in inhibiting PI3K and Cyclin D protein. Methods: Human oral squamous carcinoma cell lines SCC-9 were used in this study. A cytotoxicity assay was performed to obtain the IC 50 value of CC ethanol extract and DX on the SCC-9 cell line. Synergism evaluation of the combination CC and DX was analyzed using CompuSyn software. ELISA and the immunofluorescent assay were performed to determine the level of PI3K and cyclin D in the SCC-9 cell line after being treated with IC 50 value of CC, IC 50 value of DX and three combinations of CC and DX [7/8 IC 50 CC + 1/8 IC 50 DX (dose 1), 6/8 IC 50 CC + 2/8 IC 50 DX (dose 2), and 4/8 IC 50 CC + 4/8 IC 50 DX (dose 3). Results: CC stem ethanol extract and DX inhibited the proliferation of SCC-9 cell lines with the IC 50 value of 133.4 µg/mL and 288.3 nM, respectively. The combination of CC and DX at dose 2 (6/8 IC 50 CC + 2/8 IC 50 DX) exhibited a high decrease in PI3K and cyclin D expression. Conclusions: The combination of C. cinerariifolium and doxorubicin synergistically declined OSCC proliferation by inhibiting PI3K and cyclin D expression.

Background: Neurodegeneration due to neurotoxicity is one of the phenomena in temporal lobe epilepsy. Experimentally, hippocampal excitotoxicity process can occur due to kainic acid exposure, especially in the CA3 area. Neuronal death, astrocyte reactivity and increased calcium also occur in hippocampal excitotoxicity, but few studies have investigated immediate effect after kainic acid exposure. The organotypic hippocampal slice culture (OHSC) is a useful model for studying the neurodegeneration process, but there are still many protocol differences. In this study, minor modifications were made in the OHSC protocol. Methods: OHSC was obtained from two healthy wild type Wistar rats aged P10. Healthy culture slices were obtained and lasted up to 10 days in vitro (DIV 10). Bath application of kainic acid for 48 hours in DIV 10 followed by observation of its initial effects on neurons, astrocytes, and calcium via the expression of MAP2, GFAP, and intracellular calcium, subsequently. Results: After 48 h of kainic acid administration, there was a significant increase in intracellular calcium (p = 0.006 < α), accompanied by a significant decrease in MAP2 (p = 0.003 < α ) and GFAP (p = 0.010 < α) expression. Conclusion: These findings suggest early neuronal and astrocyte damage at the initial onset of hippocampal injury. This implies that astrocyte damage occurs early before an increase in GFAP that characterizes reactive astrogliosis found in other studies. Damage to neurons and astrocytes may be associated with increased intracellular calcium. It is necessary to develop further research regarding expression of calcium, MAP2, and GFAP at a spatial time after exposure to kainic acid and strategies to reduce damage caused by excitotoxicity.

With widespread users across the globe, topic on the impact of mobile phone in particular regarding its electromagnetic wave radiation deserves a priority. Carcinogenic effects of the electromagnetic wave radiation have been suggested by multiple studies. Effects of the radiation are to involve thermal and non-thermal pathways. Among human organs, thyroid gland is susceptible to the radiation exposure due to its location being superficial and close to the radiation transmitter. Previous studies have indicated the correlation between the mobile phone radiation and thyroid dysfunctions. However, a conclusion that the electromagnetic wave radiation is the causative agent of the thyroid dysfunctions has not been reached. Most of the study are unable to remove the influence of the external electromagnetic wave radiation. Hence, this study aimed to perform an investigation with true experimental design using animal model in an anechoic chamber – an isolated chamber which could minimize the electromagnetic wave noise from the outside. In this article, we present the description of the anechoic chamber along with the research protocols. Parameters affecting the investigation, including signal frequencies inside the chamber and specific absorption rate are also measured.

With widespread users across the globe, topic on the impact of mobile phone in particular regarding its electromagnetic wave radiation deserves a priority. Carcinogenic effects of the electromagnetic wave radiation have been suggested by multiple studies. Effects of the radiation are to involve thermal and non-thermal pathways. Among human organs, thyroid gland is susceptible to the radiation exposure due to its location being superficial and close to the radiation transmitter. Previous studies have indicated the correlation between the mobile phone radiation and thyroid dysfunctions. However, a conclusion that the electromagnetic wave radiation is the causative agent of the thyroid dysfunctions has not been reached. Most of the study are unable to remove the influence of the external electromagnetic wave radiation. Hence, this study aimed to perform an investigation with true experimental design using animal model in an anechoic chamber – an isolated chamber which could minimize the electromagnetic wave noise from the outside. In this article, we present the description of the anechoic chamber along with the research protocols. Parameters affecting the investigation, including signal frequencies inside the chamber and specific absorption rate are also measured.

Background: Neurodegeneration due to neurotoxicity is one of the phenomena in temporal lobe epilepsy. Experimentally, hippocampal excitotoxicity process can occur due to kainic acid exposure, especially in the CA3 area. Neuronal death, astrocyte reactivity and increased calcium also occur in hippocampal excitotoxicity, but few studies have investigated immediate effect after kainic acid exposure. The organotypic hippocampal slice culture (OHSC) is a useful model for studying the neurodegeneration process, but there are still many protocol differences. In this study, minor modifications were made in the OHSC protocol. Methods: OHSC was obtained from two healthy wild type Wistar rats aged P10. Healthy culture slices were obtained and lasted up to 10 days in vitro (DIV 10). Bath application of kainic acid for 48 hours in DIV 10 followed by observation of its initial effects on neurons, astrocytes, and calcium via the expression of MAP2, GFAP, and intracellular calcium, subsequently. Results: After 48 h of kainic acid administration, there was a significant increase in intracellular calcium (p = 0.006 < α), accompanied by a significant decrease in MAP2 (p = 0.003 < α ) and GFAP (p = 0.010 < α) expression. Conclusion: These findings suggest early neuronal and astrocyte damage at the initial onset of hippocampal injury. This implies that astrocyte damage occurs early before an increase in GFAP that characterizes reactive astrogliosis found in other studies. Damage to neurons and astrocytes may be associated with increased intracellular calcium. It is necessary to develop further research regarding expression of calcium, MAP2, and GFAP at a spatial time after exposure to kainic acid and strategies to reduce damage caused by excitotoxicity.

Background: Tourniquet is a common instrument used in Orthopedics field to reduce blood loss, providing a better operating field. One of the deleterious effects of tourniquet is ischemic-reperfusion injury, which occurs after deflation of the tourniquet. One of the possible ways to mitigate ischemic-reperfusion injury is by administrating antioxidants to reduce oxidative stress. Natural ingredients like Aloe vera are well known for its antioxidant and anti-inflammatory activities. This experimental study was conducted by fracturing the tibia of male Wistar strain rats ( Rattus norvegicus) and administering Aloe vera gel orally as antioxidant. Method: A total of 18 rats were used in this study, divided into 6 groups. The first group was the control group, that included rats with fractured tibia not receiving torniquet and aloe vera treatment. The second and third groups were rats with fracture, who received torniquet application for one hour and two hours respectively without aloe vera treatment. The rest were the treatment groups, the rats with factures were given different dosages of Aloe vera extract: 40 mg/kg bodyweight (BW), 60 mg/kg BW, and 80 mg/kg BW. Three hours after administration, the tourniquet was inflated for two hours, followed by its deflation. The tibia was harvested to examine levels of superoxide dismutase (SOD), Malondialdehyde (MDA), and Bone morphogenetic protein 7 (BMP-7), and callus diameter, and osteoblast numbers were evaluated. Result: Administration of Aloe vera extract reduced the oxidative stress parameters (SOD and MDA). Tourniquet application decreased the osteoblast cell count and callus diameter, and administration of Aloe vera extract increased both variables close to the control value (p Conclusion: The result of this study suggests that Aloe vera extract could be used to ameliorate ischemic-reperfusion injury.