Molecular Exploration of Biomarkers as Early Warning System of Aquatic Pollution

LAS exposure to potentially cause pollution in aquatic environments. LAS Content in the waters, harmful to aquatic organisms and humans. Biomarkers required as "early warning" of pollution on aquatic biomonitoring program. Biomarkers can provide a picture of the effect of changes in environmental quality. The response to the stress caused by LAS is used as a biomarker of pollution and is expected to provide an overview effect on humans in the future. This study analyzes the differences in molecular changes in the hepatocyte Cyprinus carpio L were exposed to various concentrations of LAS. LAS as stressors created by the concentration of 0.01, 0.02; 0.03; 0.04; 0.05 mg / l with a long exposure of 24 hours, 48 hours, 72 hours, 96 hours, and 8 days. Biomarker expression in hepatocyte observed using immunohistochemical methods. The results obtained were analyzed statistically using the 2-way MANOVA. The results showed a significant level of p = 0.0005 on HSP70 protein expression, iNOS, p38 MAPK, and CYP 1A compared to the control group. Increased concentrations of LAS and the duration of exposure resulted in increased number expressing hepatocyte biomarker with a significant level (p = 0.0005). The conclusion of this study is that the expression of HSP70, iNOS, p38 MAPK, and CYP 1A can be used as a biomarker of pollution in aquatic environments LAS. Keywords: Aquatic,  biomarkers, early warning system, molecular, pollution

Anticancer Effects of LBA-ST Yogurt Supernatant on HeLa Cells via Heat Shock Protein 27 Decrease In Vitro

Heat Shock Protein 27 (Hsp27) is overexpressed in cervical cancer as a response to stress conditions. Hsp27 overexpression increase invasion, migration, and adhesion pathways of cancer cells. The Yogurt supernatant contains Short-Chain Fatty Acids (SCFA) include acetate, lactate, and butyrate which have anticancer activity. This study aimed to investigate supernatant of LBA-ST (Lactobacillusbulgaricus-acidophilus, Streptococcusthermophillus) Yogurt can decrease the expression of Hsp27 in HeLa culture cells. The mechanism on how supernatant yogurt inhibit invasion, migration, and adhesion was studied by immunocytochemistry. The data was then collected and analyzed using One-Way ANOVA. From this study, it can be concluded that the expression of proteins that play a role in invasion, adhesion, and migration of the Hsp27 was proven to be decreased (p< 0.05 ± 0.005).Keywords: HeLa cells, yogurt supernatant, Lactobacillus bulgaricus-acidophilus, Streptococcus thermophillus, Hsp2

In silico and in vivo anti-inflammatory studies of curcuminoids, turmeric extract with zinc oxide, and eugenol

Purpose: To determine the anti-inflammatory activity of curcuminoids in comparison with that of eugenol in silico, and to determine the anti-inflammatory activity of wound dressings made from zinc oxide powder and liquid turmeric extract with a high curcuminoid content.Methods:  In silico studies were conducted, using Molegro Virtual Docker program, to predict the antiinflammatory potency of curcuminoids (curcumin, demethoxycurcumin, bisdemethoxycurcumin) and eugenol against COX-2 receptors. In vivo studies to evaluate anti-inflammatory activity via TNFα expression, were carried out using thirty Wistar rats as subjects, divided into two groups: A (sacrificed on day 3) and B (sacrificed on day 7). Each group contained three subgroups (n = 5): A1, B1 were excised without a dressing as control subgroups; A2, B2 were excised followed by the application of zinc oxide with a turmeric extract dressing; and A3, B3 were excised followed by the application of zinc oxide with an eugenol dressing.Results: The in silico studies confirmed the anti-inflammatory activity of curcumin (-132.905 kcal/mol), demethoxycurcumin (-130.265 kcal/mol), bisdemethoxycurcumin (-118.827 kcal/mol) in relation to the COX-2 receptor to be greater than that of eugenol (-78.718 kcal/mol). The in vivo studies of TNFαexpression showed that the levels of activity in the groups without dressings were significantly higher than in those with dressing (p&lt;0.05), while the lowest TNFα expression were for zinc oxide with turmeric extract dressings.Conclusion: The combination of zinc oxide with turmeric liquid extract has a higher anti-inflammatory effect than eugenol as demonstrated by both in vivo and in silico studies. This combination can, therefore, be used as an alternative to zinc oxide eugenol wound dressings.Keywords: Anti-inflammatory, Curcuminoids, Turmeric, Zinc oxide, Eugenol, Wound dressing, In silico, TNF

Protective Effects of Propolis Extract in a Rat Model of Traumatic Brain Injury via Hsp70 Induction

BACKGROUND: Traumatic brain injury (TBI) is one of the major global health problems. Secondary brain injury is a complex inflammation cascades process that causes brain cell apoptosis. Propolis is a natural product that has neuroprotective property. AIM: This study aimed to investigate the effect of propolis toward Hsp70 expression with apoptosis marker in brain tissue after TBI. METHODS: Thirty-three Sprague Dawley rats were randomised into three treatments group, i.e. sham-operated controls, closed head injury (CHI), and CHI with propolis extract (treatment group). In the treatment group, propolis was given 200 mg/kg per oral for 7 days then harvested brain tissues after sacrificed by cervical dislocation at day 8. We investigated Hsp70, Caspase 3, apoptosis-inducing factor (AIF), and TUNEL assay expression using immunohistochemistry staining. Statistical test using one-way ANOVA test and Tukey HSD as post hoc test. RESULTS: Mean of positive Hsp70 stained cells in group 1 was 6.82 ± 2.14, group 2 was 3.91 ± 2.26, and group 3 was 9.64 ± 3.53 with a significant difference of Hsp70 expression distribution within groups (p = 0.0001). Mean of positive caspase 3 stained cells in group 1 was 5.45 ± 2.30, group 2 was 13.82 ± 2.44, and group 3 was 7.03 ± 1.54 with a significant difference of caspase3 expression distribution within groups (p=0.0001). Mean of positive AIF stained cells in group 1 was 5.36 ± 2.11, group 2 was 12.82 ± 1.40, and group 3 was 8.09 ± 1.81 with a significant difference of AIF expression distribution within groups (p = 0.0001). Mean of positive TUNEL assay stained cells in group 1 was 4.82 ± 2.04, group 2 was 11.55 ± 1.51, and group 3 was 7.64 ± 1.96 with a significant difference of TUNEL test expression distribution within groups (p = 0.0001). CONCLUSION: Propolis may protect brain cell from apoptosis after injury by maintaining Hsp70 expression in addition to antioxidant and anti-inflammatory

CURCUMIN PREVENTS COCHLEAR OXIDATIVE DAMAGE AFTER NOISE EXPOSURE

Objective: To demonstrate curcumin as the safe and effective therapeutic agent in the prevention and treatment of oxidative damage in fibroblasts within the cochlear supporting tissues and lateral wall following noise exposure by neutralizing oxidative stress-inducing agents, such as malondialdehyde (MDA) and hydrogen peroxide (H2O2).Methods: Twenty-four Rattus norvegicus were randomly divided into 4 groups (n = 6). Group 1: The control group; group 2: noise (+); group 3: noise (+), 50 mg/day curcumin (+); group 4: noise (+), 100 mg/day curcumin (+). All groups (except group 1) were subjected to 100 dB SPL for 2 h per day for 14 d. Curcumin used in this study was derived from Curcuma longa L. (Turmeric) with curcumin [28.1±1.0]% w/w compared to Standard and administered orally for 14 d. All samples were Immuno histo-chemistrically examined for the expressions of MDA in cochlear fibroblasts and colorimetrically examined for H2O2 levels in cochlear tissues using the colorimetric reader.Results: The results obtained showed significant differences for the expressions of MDA (P&lt;0.05) in all groups, and significant differences for H2O2 levels (P&lt;0.05) in all groups, except in group 1 compared to 4 and group 3 compared to 4.Conclusion: Curcumin proved to be potentially effective in the prevention and treatment of oxidative damage in fibroblasts within the cochlear supporting tissues and lateral wall following noise exposure by decreasing the expressions of MDA and H2O2 levels.Â

Efficacy of Neuroprotection from Curcumin through Heat Shock Protein 70 Induction in Traumatic Brain Injury – Rat Model

BACKGROUND: Traumatic brain injury (TBI) is the most common problem that caused morbidity and mortality in the world. Secondary brain injury is a complex cascade that causes brain cell apoptosis. Curcumin is a natural product that has neuroprotective properties. AIM: This study aimed to investigate the effect of curcumin toward heat shock protein 70 (HSP 70) expression against the expression apoptosis marker (apoptosis-inducing factor [AIF], caspase-3, and TUNEL assay) in brain tissue after TBI. METHODS: Thirty-three Sprague Dawley rats were randomized into three treatment groups, that is, sham-operated controls, closed head trauma (CHT), and CHT with curcumin extract (treatment group). In the treatment group, curcumin was given 500 mg/kg per oral for 7 days, then brain tissues were investigated (marker AIF, caspase-3, TUNEL assay, and HSP 70) through immunohistochemistry. Statistical test using one-way ANOVA test and Tukey honestly significant difference as post hoc test. RESULTS: The mean of positive AIF stained cells in Group A was 5.36 ± 2.11, Group B was 12.82 ± 1.40, and Group C was 3.82 ± 1.40, with a significant difference of AIF expression between Groups C and B (p &lt; 0.05). Mean of positive caspase-3 stained cells in Group A was 5.45 ± 2.30, Group B was 13.82 ± 2.44, and Group C was 3.82 ± 1.54, with a significant difference of caspase-3 expression between Groups C and B (p &lt; 0.05). Mean of positive TUNEL assay stained cells in Group A was 4.82 ± 2.04, Group B was 11.55 ± 1.51, and Group C was 3.55 ± 1.70, with a significant difference between Groups C and B (p &lt; 0.05). Mean of positive HSP 70 stained cells in Group A was 6.82 ± 2.14, Group B was 3.91 ± 2.26, and Group C was 10.27 ± 2.45 with a significant difference of HSP 70 expression distribution within groups (p &lt; 0.05). CONCLUSION: Curcumin may protect brain cells from apoptosis after close head trauma by upregulated HSP 70 expression

Expression of Osterix and SOX9 after Administration of Gourami Fish Scale Collagen in Wistar Rats

The current goal of periodontal treatments is to regenerate periodontal tissue by adding bone grafts. Various materials are being explored as bone substitutes, including type 1 collagen. Gourami fish scales are known to be an alternative source of type 1 collagen and have potential as a substitute material for bone grafts. The objective of this study was to determine whether there is an increase in the expression of Osterix (OSX) and SOX9 after the administration of gourami (Osphronemus goramy) fish scale collagen in extracted socket Wistar rats (Rattus norvegicus). The research was conducted on 32 male Wistar rats that were classified into four groups: control groups on day 7 (K7) and day 14 (K14); and treatment groups on day 7 (P7) and day 14 (P14). The expression of OSX and SOX9 was visualized by immunohistochemical staining of bone tissue preparations by anti-OSX and SOX9 monoclonal antibodies. The ANOVA results for OSX expression had a significant difference in OSX expression between that in the control and treatment groups on day 7 and day 14. The ANOVA results for SOX9 expression on day 7 had a significant difference in SOX9 expression between that in the control and treatment groups. ANOVA results for SOX9 expression on day 14 had no significant difference in the expression of SOX9 between the treatment and control groups. The conclusion of this study was administration of Gourami scale collagen can increase the expression of OSX and SOX9 in socket’s Wistar rats

Analysis of the Histopathology, TNF-α of Microglia Cells Expression, NRG-1/erbB Oligodendrocyte, and Ki67/Apoptosis of Dentate Gyrus Rattus novergicus Brain After Acute Traumatic Brain Injury

Head trauma or traumatic brain injury (TBI) gives most serious impact on the central nervous system. Several experimental models have been established to mimic different pathogenesis characteristics of TBI. The purpose of this study was to determine whether there is evidence of hystopathological lesions in the brain tissue after Marmorou TBI models. This study uses Rattus norvegicus Sprague Dawley strain. Macroscopic and microscopic observations on the brain tissue were done. Macroscopic lesions were observed in the brain. Microscopic observation was performed with Haematoxylin-Eosin (HE) staining and immunohistochemistry on the distribution of microglia cells and pyramidal cells in the cortex. Meanwhile, the distribution of NRG-1/ErbB, proliferation, and apoptosis were observed in the hippocampus. The results of macroscopic observation showed that there were wounds caused by falling loads and vasodilatation. On microscopic observation, the TBI group showed an increase in neutrophils distribution and distribution of activated microglia to produce TNF-α, and decrease in the number of cortical pyramidal cells significantly. The distribution of NRG-1 tended to decrease after exposure of TBI and had no effect on its receptor, erbB. Exposure of TBI appears to lower the activity of neuronal cells proliferation in dentate gyrus (DG) area and significantly increase the number of apoptotic cells. Marmarou model is a physiological model of TBI that spontaneously occurs following a trauma to the head, for example trauma due to an accident. This data can be used as a preliminary data of inflammation and tissue regeneration of disrupted adult brain. Therefore, this research could be used as the basis in the studies of therapeutic agents in the process of neurogenesis of brain cells.Keywords: traumatic brain injury, ERG-1/ErbB, dentate gyrus, Ki67, TNF-a, microgli

Spade Leaf Extract Phytosome Modulates Krox-20, Neuregulinـ1, Phospholipids, and Cognitive Function of Traumatic Brain Injury Model in Rats

Traumatic brain injury (TBI) is a disorder of the central nervous system due to head trauma. TBI can damage nerve membrane phospholipids and decrease protein synthesis of neuregulin-1 (NRG-1) because of the transcription factor Krox-20. These conditions cause the lowering in nerve re-myelination which contribute to the decline of cognitive function. In Indonesia, citicoline is a neuroprotective drug that widely used to repair and prevent further damage of the nerve cells membrane caused by trauma. Spade leaf (Centella asiatica) extract phytosome (SEP) is a model of drug delivery system which expected to enhance the therapeutic effects as neuroprotective drug. This study aims to demonstrate and compare the effectiveness from SEP and citicoline as a neuroprotective characterized by increasing the activation of Krox-20, the expression of NRG-1, the distribution of phospholipids, and the improvement of cognitive levels on TBI-induced rats. Rats were divided into 5 groups namely: control (-); control (+); treatment with SEP 90mg/kgBW; citicoline 250mg/kgBW, and SEP in combination with citicoline. Krox-20, NRG-1, and phospholipids expression were measured by immunohistochemical assay, while cognitive function were assessed with the Morris Water Maze test. According to ANOVA test results, it was showed that SEP improved the nerve cells through the activation of Krox-20, NRG-1 expression, and distribution of phospholipids significantly (p <0.05). Based on Morris Water Maze test, SEP also improved the cognitive function in TBI-induced rats. Thus, it can be concluded that spade leaf extract phytosome combined with citicoline increase a higher phospholipids distribution and give the fastest time in the cognitive tests compared with of spade leaf extract phytosome and citicoline alone.Keywords : NRG-1, Krox-20, phospholipids, phytosome, Centella asiatica

Immunohistochemical analysis of stem cells from human exfoliated deciduous teeth in carbonate apatite scaffold for the alveolar bone defect in Wistar rats (Rattus Novergicus)

Background: Stem cells from human exfoliated deciduous teeth (SHED) seeded in carbonate apatite scaffold (CAS) may have multiple functions that could be used to regenerate the alveolar bone defects. The purpose of this study is to examine the ability of SHED and CAS in alveolar bone defects using an immunohistochemical analysis. Methods: ten three-month-old healthy male Wistar rats (R. novergicus) that weighed between 150–250 grams (g) were used as animal models. A simple blind random sampling method was used to select the sample that was assigned to the study group for CAS and SHED seeded in CAS (n=5). The animal study model of the alveolar bone was established by extracting the anterior mandible teeth. Rodent anesthesia was applied to relieve the pain during the procedure for all test animals. Immunohistochemistry was performed after seven days to facilitate the examination of the receptor activator of NF-κβ ligand (RANKL), osteoprotegrin (OPG), transforming growth factor-β (TGF-β), vascular endothelial growth factor (VEGF), runt-related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), osteocalcin, and osteopontin expression. The data was analyzed using the unpaired ttest (p<0.01) and Pearson’s correlation test (p<0.05). Results: The OPG, RUNX2, TGF-β, VEGF, ALP, osteocalcin, and ostepontin expressions were higher in SHED seeded in CAS than CAS only with a significant difference between the groups (p<0.01). Furthermore, the RANKL expression was lower in SHED seeded in CAS compared to CAS only. There was a strong reverse significant correlation between OPG and RANKL expression (p<0.05). Conclusions: The number of osteogenic marker expressing cells, suc