Effect of Centella asiatica ethanol extract on zebrafish larvae ( Danio rerio) insomnia model through inhibition of Orexin, ERK, Akt and p38.

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.

sCD163, sCD28, sCD80, and sCTLA-4 as soluble marker candidates for detecting immunosenescence.

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.

Nicotine is associated with smoking dependence and vascular inflammation through cotinine: A mediation analysis.

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<0.001) regarding the cycle formed with decreased NO levels as a result of damage caused by Tie2-mediated inflammation. CONCLUSIONS: There is a relationship between NO levels and cotinine through nAChRs, which mediate the nicotine dependence mechanism and Tie2 expression.

Effects of mobile phone electromagnetic radiation on thyroid glands and hormones in Rattus norvegicus brain: An analysis of thyroid function, reactive oxygen species, and monocarboxylate transporter 8.

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.

Effect of epigallocatechin gallate in green tea on preventing lens opacity and αB-crystallin aggregation in rat model of diabetes.

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<0.05. EGCG exposure can significantly lower RBS with an R 2 of 0.5634 (56.34%). The same analysis on EGCG exposure and the degree of lens opacity obtained a significance level of P<0.05 and increased exposure to EGCG can significantly lower the degree of lens opacity with an R 2 of 0.8577 (85.77%). Correlation analysis between EGCG and the expression of lens αB-crystallin can be concluded that the higher the EGCG exposure administered, the higher the native lens αB-crystallin expression and the lower the aggregate lens αB-crystallin expression. There was also significant effect in which every 1 mg/kg body weight dose of EGCG can increase the native lens αB-crystallin expression by 0.0063 and decrease the aggregate lens αB-crystallin expression by 0.0076. CONCLUSION: The administration of EGCG at a dose of 300, 600, and 1200 mg shows a significant effect on preventing lens opacity and aggregation of αB-crystallin in diabetic rat models and this research could be a biomolecular prevention of cataract.

The Effect of Salmonella AvrA Infection on Colon Cancer: An in-silico Study.

OBJECTIVE: Colorectal cancer (CRC) is one of the main causes of morbidity and mortality due to cancer. The purpose of this in-silico study was to examine the relationship of chronic infection mechanisms caused by Salmonella Anti virulence agent A (AvrA) to gene mutations in the carcinogenic process of CRC. METHODS: Gene expression data on the mouse colon was obtained from the GSE22215 dataset | Gene Expression Omnibus (GEO). Adjusted p-value was calculated using Benjamini & Hochberg False Discovery Rate (FDR<0.01). Gene expression in colon adenocarcicoma tumors was obtained from The Cancer Genome Atlas's (TCGA) Genomic Data Commons (GDC) dataset containing 458 colon tumor samples. RESULT: Expressions of MLH1, MSH2, EPCAM, APC, and PMS2 in cases of colon adenocarcinoma tumor showed a correlation with genes that underwent changes due to Salmonella AvrA infection. Among the gens of interest, EPCAM was the gene that had the highest correlation compared to other genes (MLH1, MSH2, APC, and PMS2) (n= 514, Gene r-p value < 0.01 =22355). There were 514 genes that had a correlation with cases of AvrA infection. Tumor Necrosis Factor (TNF), which is a gene that is upregulated in AvrA infection and correlates negatively with EPCAM, had the highest BC value compared to other gens (p= 0.0000768). Survival probability showed that EPCAM was highly expressed and it can increase survival time. In addition to TNF, our study indicated that IL1B (p= 0.000419), S100A8 (p= 2.02E-05), S100A9 (p=0.000419) correlated with the gene of interest. CONCLUSION: Late Salmonella AvrA infection affects the expression of genes involved in inflammation in colorectal cancer samples.

The Effect of Citicoline on the Expression of Matrix Metalloproteinase-2 (MMP-2), Transforming Growth Factor-ß1 (TGF-ß1), and Ki-67, and on the Thickness of Scleral Tissue of Rat Myopia Model.

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-ß1, 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-ß1, 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-ß1 expression. Citicoline is able to decrease MMP-2 expression and fibroblast proliferation and increase TGF-ß1 expression and scleral tissue thickness significantly in the scleral tissue of rat models for myopia.

Detection of Vascular Inflammation and Oxidative Stress by Cotinine in Smokers: Measured Through Interleukin-6 and Superoxide Dismutase.

Purpose: Smoking is a significant risk factor in developing cardiovascular disease pathogenesis through oxidative stress and inflammation mechanisms. This study used cotinine as a biomarker of nicotine exposure levels in the body, which was associated with levels of Interleukin-6 (IL-6) and Superoxide Dismutase (SOD) as markers of oxidative stress and vascular inflammation. The research aimed to analyze the effect of cotinine levels on the expression of IL-6 and SOD. Methods: This study used a cross-sectional design on 200 subjects, consisting 100 smokers and 100 non-smokers. Cotinine levels, IL-6 expression, and SOD were measured from the blood serum of each subject using the Enzyme-Linked Immunosorbent Assay (ELISA) method. Then the data were analyzed using Generalized Structured Component Analysis (GSCA). Results: There was a significant effect of cotinine levels on the reduction of SOD mediated by IL-6 (CR = 4.006). Cotinine levels also increased IL-6 mediated by SOD (CR = 4.292). The structural model shows that higher cotinine levels will increase IL-6 expression, and conversely, SOD expression will decrease. Conclusion: High cotinine levels cause an increase in the inflammatory process and oxidative stress in the vasculature of smokers, which is characterized by high IL-6 expression and low SOD expression.

IMMUNE RESPONSE UPON THE ADMINISTRATION OF RECOMBINANT PROTEIN ANTIBODIES Ag-38 KDa Mycobacterium tuberculosis AND RIFAMPICIN EX-VIVO.

Background: Development a granuloma model resembling latent tuberculosis in vitro is needed with a fast and efficient time to be used as an effective therapy. This study aimed to form efficient granulomas, increase cellular immunity and humoral immunity, and evaluate growth on media using recombinant protein antibody Ag38kDa, Rifampicin, and a combination of both. Peripheral Blood Mononuclear Cell (PBMC) in vitro is derived from a healthy individual separated from monocytes and lymphocytes. Materials and methods: Monocytes are matured into macrophages and then combined macrophages and lymphocytes to the Roswell Park Memorial Institute (RPMI) medium. Flow cytometry analysis was used to count the number of cells, and cytokine levels were measured using ELISA. The result from the treatment was planted on the Lowenstein-Jensen medium. Results: Granulomas-like aggregates was formed after one-day post-infection with Mycobacterium tuberculosis (M.tb). A significant increase in immune response occurred in the number of macrophages, Th1, and Tregs in the combination group compared to the Mtb infection group. The number of Th2 and Th17 cells in the combination group was compared with the control but not significantly. TNF-α cytokine levels increased in the combination group compared to Mtb infection, while in IL-4, we found between all groups, there was no significant difference. Bacterial colonies on culture in the Lowenstein-Jensen medium were only seen in positive controls. Conclusion: Our study concluded that administration of a combination between Ag38kDa recombinant antibody and rifampicin could inhibit granuloma formation and enhance immune response.

Shigella flexneri vaccine development: Oral administration of peptides derived from the 49.8 kDa pili protein subunit activates the intestinal immune response in mice.

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 ß-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 ß-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.

Comparative performance of ELISA and dot blot assay for TSH-receptor antibody detection in Graves' disease.

BACKGROUND: Graves' disease (GD) is an autoimmune disease, and it accounts for major cases of hyperthyroidism. Antibody against thyroid-stimulating hormone receptor/TSHR (TRAb) is responsible for hyperthyroidism and is considered as a diagnostic marker for GD. Therefore, we developed a recombinant protein of human TSHR-169 (hTSHR-169), which was specifically recognized TRAb in the serum of GD patients and then compare the diagnostic performance between ELISA and dot blot of TRAb tests for their ability to diagnose GD. METHODS: 20 GD patients and 20 healthy individuals from the Indonesian population were enrolled. TRAb concentration and density were quantified. Comparative analysis was performed using receiver-operating curve (ROC) analysis. RESULTS: For dot blot assay, the minimum concentration to detect TRAb requiring 100 ng of antigen with antiserum diluted at 1:60. For diagnosing GD, the ELISA yielded a higher AUC compared with the dot blot assay (0.95 and 0.85, respectively). Using the recommended cutoff values, the efficiency of both assays was examined by comparing the specificity and sensitivity of the assays to the clinical diagnosis. The ELISA showed 80% and 95%, while the dot blot assay showed 70% and 95% sensitivity and specificity, respectively. CONCLUSION: Although the dot blot assay exhibited lower performance than the ELISA method, the dot blot assay is a simple and rapid diagnostic assay that is suitable for diagnosing GD in rural areas, in which healthcare facilities sometimes are not accessible.

Association of CDH13 Gene Polymorphism and Metabolic Syndrome in Gambian Population.

BACKGROUND: Polymorphism in CDH13 gene, which encodes for the adiponectin receptor, T-cadherin, is a genetic risk factor associated with metabolic syndrome. CDH13 rs3865188, which is found in the promoter region of the CDH13 gene, has been found to be associated with metabolic syndrome and its traits in Asian and European Caucasian populations. However, to the best of our knowledge, it was yet to be assessed in a Black African population. OBJECTIVE: The aim of this study was to investigate the association of CHD13 rs3865188 and metabolic syndrome in a Gambian population. METHODS: It was a genetic association study in a cross-sectional design in 136 Gambian participants. CDH13 rs3865188 was genotyped using PCR master mix and sequencing. Blood sugar, triglyceride and high-density lipoprotein levels were determined by standard clinical laboratory methods. RESULTS: CDH13 rs3865188 was found to be significantly associated metabolic syndrome (p=0.034). Genotype AT appeared to be risk factor for metabolic syndrome (OR=2.41, 95% CI, 1.20-4.84, p=0.014). We found genotypes CC and CA in CHD13 rs3865188 for the first time. CONCLUSION: Our study demonstrated significant association between CDH13 rs385618 and metabolic syndrome in a Gambian population (Black African population for the first time). Individuals with genotype AT are at higher risk of developing metabolic syndrome.

Prevalence of Metabolic Syndrome and its Components in Kanifing Municipality, The Gambia.

BACKGROUND: Metabolic syndrome (MetS) is a complex syndrome with clustering of interrelated risk factors for cardiovascular disease and diabetes. Its rising worldwide prevalence has been largely related to the increasing obesity. In The Gambia, the last and only time a MetS related study was conducted, and then reported, was 21 years. Therefore, there is need for evaluating the prevalence of MetS and its components in the country. OBJECTIVE: This study was aimed to evaluate the prevalence of MetS and its individual components in Kanifing Municipality (KM). METHODS: It was a cross-sectional study conducted at Kanifing General Hospital, Kanifing Municipality. Data obtained from each participants included anthropometric indices, blood pressure, fasting plasma glucose, triglyceride and high-density lipoprotein levels, and clinical information. RESULTS: One hundred and thirty-six participants were included in the analysis. The overall MetS prevalence was 54.4% with significant female predominance (female, 58%; male, 29.4%; P=0.025). The most predominant component among the study population was central obesity (raised WC) (72.8%). Hypertriglyceridemia was found to be the strongest predictor of MetS among our participants (OR: 118.13; 95% CI: 33.79-412.77; P < 0.001). CONCLUSION: Our study discloses a very high prevalence of MetS among the participants, and a significant female predominance, with central obesity the commonest Mets component. The results suggest that hypertriglyceridemia is the strongest predictor of metabolic syndrome in our study participants.

Extra Virgin Olive Oil Modulates Vasodilator Enzyme Level by Repairing Angiogenesis Function in Rat Model of Preeclampsia.

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.

Reperfusion Interval as a Prevention of Lung Injury Due to Limb Ischemia-Reperfusion After Application of Tourniquet in Murine Experimental Study.

BACKGROUND: Tourniquet use is prevalent in the orthopaedic field to achieve a bloodless operating field, but it poses risks of local and systemic complications, including lung injury. This study aims to examine the effect of tourniquet application on the hindlimb of a rat to its lung. MATERIALS AND METHODS: This is an experimental study with 48 male Wistar strain rats as samples. The rats were divided into group A (n = 24), killed directly after fracturization and tourniquet application, and group B (n = 24), killed 14 days post-procedure. Each group was divided into four: group A1/B1 (control group, three hours tourniquet application without reperfusion interval), A2/B2 (5-min reperfusion between 2-h and 1-h tourniquet application), A3/B3 (10-min reperfusion), and A4/B4 (15-min reperfusion). The lung tissue was examined histologically within ten high-power fields (400 × magnification). The severity of lung injury was measured using the Lung Injury Score (LIS). The oxidative damage was measured by determining the malondialdehyde (MDA) level, using the TBARS (thiobarbituric acid reactive substance assay) method. RESULTS: There was a dose-dependent decrease of LIS and MDA in groups A and B with increasing reperfusion interval. Fifteen-minute reperfusion interval caused a 54.55% and 45.33% LIS reduction in groups A and B, respectively. All pair-wise group comparisons (p < 0.05) showed significant differences. Five-minute interval reduced the MDA level by 16.56% and 30.13% in groups A and B, respectively. All possible pair-wise comparisons in both groups A and B also showed a significant difference (p < 0.05). CONCLUSIONS: Reperfusion interval is a possible clinical approach to mitigate the remote organ damage induced by limb ischemia-reperfusion injury.

Preparation and biodegradable properties of hydroxyapatite nanoparticle composite coated with poly lactic-co-glycolic acid/polyvinyl alcohol for bone regeneration.

BACKGROUND: Bone loss rapidly increases 6 months post tooth extraction, which causes the atrophy of the alveolar bone. Two kinds of biomaterials which can stimulate bone regeneration are bioceramics and polymers. Making a composite of biomaterials results in better physical and biomolecular characteristics in comparison with a bioceramic or a polymer alone. Hydroxyapatite nanoparticles (HANPs) are one of the bioceramics commonly used for bone regeneration; they can degrade faster than hydroxyapatite (HA) microparticles, but have an insufficient pore size. Polyvinyl alcohol (PVA) and poly lactic-co-glycolic acid (PLGA) are polymers which have been used for biomedical applications. However, PLGA alone has insufficient cell attachment and PVA alone slowly degrades in the bone tissue. OBJECTIVES: The aim of the present study was to analyze the biodegradation properties of the HANP/PLGA/PVA composites and investigate the pore size. MATERIAL AND METHODS: The HANP/PLGA/PVA composites were prepared using the freeze-drying method, with 20% (w/w) of HANP and 20% (w/w) of PLGA. Morphology and the pore size were determined by means of the field emission scanning electron microscopy (FE-SEM) analysis. Biodegradation properties were determined by calculating water uptake and water loss for 1, 3 and 6 weeks. Statistical analysis was performed based on the one-way analysis of variance (ANOVA) at p < 0.05. RESULTS: The HANP/PLGA/PVA composites had the greatest mean pore size and a rougher surface than others (176.00 ±61.93 µm; p < 0.05). Moreover, the HANP/PLGA/PVA composites had the greatest water uptake, significantly in the 3rd (730.46%; p < 0.05) and 6th weeks (731.07%; p < 0.05), and water loss in the 6th week (67.69%; p < 0.05). CONCLUSIONS: The HANP/PLGA/PVA composites have optimal pore size, morphology and degradability, which shows their high potential as an effective bone scaffold to repair the alveolar defect post tooth extraction.

Pro-apoptotic and anti-proliferative effects of Physalis angulata leaf extract on retinoblastoma cells.

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.

Autistic Children Are More Responsive to Tactile Sensory Stimulus.

OBJECTIVE: This research was an experimental study that was aimed to detect differences response of tactile sensory stimulus between normal children and children with sensory brain development disorders such as Autism Spectrum Disorder (ASD). MATERIALS & METHODS: A total of 134 children, in two groups including 67 healthy children (control) and 67 children with autism were studied. Tactile sensory stimulus responses in children were tested directly using a Reflex Hammer. In addition, tactile sensory sensitivity was also assessed via questionnaire Short Sensory Profile (SSP) filled out by the child's parents. All response data were analyzed using Fisher's Exact Test; questionnaire data was analyzed with the Mann-Whitney U Test. RESULTS: Autistic children were more sensitive to palpation and pain than children who were not autistic. Furthermore, the value of SSP was also significantly higher (P<0.05) in autistic children, which means that they always responded to all categories in the SSP questionnaire than children who are not autistic. CONCLUSION: Autistic children are more sensitive to tactile sensory stimulus and all categories of SSP than children who are not autistic.

Genistein modulates the estrogen receptor and suppresses angiogenesis and inflammation in the murine model of peritoneal endometriosis.

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-ß, TNF-α, IL-6, VEGF, and HIF-1α were performed immunohistochemically. Expression of estrogen receptor-α, estrogen receptor-ß, TNF-α, IL-6, VEGF and HIF-1α increased significantly compared with the control group (p < 0.05). All doses of genistein decreased the expression of estrogen receptor-α, increased estrogen receptor-ß, lowered VEGF and HIF-1α significantly compared with endometriosis 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-ß 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.

Mechanism of retinal pericyte migration through Angiopoietin/Tie-2 signaling pathway on diabetic rats.

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<0.05). Pericyte number reduced by 40% (1:2.4) after 5wk diabetes on diabetic rats. The pericyte: endothelial ratio on group with Tie-2 inhibitor were 1:1.8. The same result shows on group with Akt/PKB inhibition. ERK1/2 inhibitor group shows the best results of pericyte: endothelial ratio (1:1.7). Inhibition on Tie-2 receptor decreased the phosphorylation activity of Tie-2, ERK1/2 and Akt/PKB pathway. ERK1/2 inhibition also decreasing the phosphorylation of Tie-2 and Akt/PKB. But on Akt/PKB inhibition, the phosphorylation of Tie-2 and ERK1/2 were relative the same. CONCLUSION: Ang-2 has a role for pericyte migration on diabetic rats through Tie-2 receptor, ERK1/2 and Akt/PKB pathways. ERK1/2 is a dominant pathway based on the ability to supress another pathway activity and decreasing pericyte migration on diabetic rats.