Supplementation of Bacillus subtilis and Lactobacillus casei to increase growth performance and immune system of catfish (Clarias gariepinus) due to Aeromonas hydrophila infection.

Background and Aim: Catfish has a high economic value and is popular among consumers. To ensure well-stocked catfish stocks, good fisheries management must also be ensured. The high demand for catfish must be supplemented by preventive measures against pathogenic bacterial infections using probiotics with high potential for Lactobacillus casei and Bacillus subtilis. The aim of this study was to determine the effect of probiotic supplementation consisting of a combination of L. casei and B. subtilis probiotics on the growth, immune system, water quality, proximate value of feed, and body composition of catfish infected with Aeromonas hydrophila. Materials and Methods: This study used a completely randomized study with eight treatments and three replications. The manipulated factor was the probiotic concentration [0% (A), 0.5% (B), 10% (C), and 15% (D)] in groups of catfish infected and uninfected with A. hydrophila. Combination of B. subtilis, and L. casei that were used in a 1:1 ratio of 108 colony forming unit/mL. The study lasted for 42 days. On the 35th day, A. hydrophila was infected by intramuscular injection into fish. The Statistical Package for the Social Sciences (SPSS) software version 23.0 (IBM SPSS Statistics) was used to analyze data on growth, immune system, and water quality. Results: Providing probiotics in feed can increase the nutritional value of feed based on proximate test results. There were significant differences in average daily gain (ADG), feed conversion ratio (FCR), and survival rate (SR) parameters in the group of catfish infected with A. hydrophila (p > 0.05); however, there were no significant differences in final body weight, specific growth rate (SGR), and percentage weight gain. Interleukin-1ß (IL-1ß) levels were significantly different between treatments C and D. The tumor necrosis factor (TNF) α parameters were significantly different between treatments A and C, whereas the phagocytic activity of treatment A was significantly different from that of treatment D. There was a significant difference (p > 0.05) in the growth parameters of SGR, ADG, and FCR in the group of fish that were not infected with A. hydrophila, with the best treatment being a probiotic concentration of 15%, but there was no significant difference in the SR parameters. IL-1ß and TNF-α levels significantly differed between E and E0 (15% probiotics) but were not significantly different in terms of phagocytosis parameters. Conclusion: Based on the results of this study, it can be concluded that using a combination of probiotics L. casei and B. subtilis can improve the growth, immune system, water quality, proximate value of feed, and body composition of catfish infected with A. hydrophila.

The Effect of G. applanatum Crude Polysaccharide Extract on Proinflammatory Cytokines and Proapoptotic Caspases in HeLa Cell Line: An In Vitro Study.

Polysaccharide extracts exhibit promise as potential anticancer agents. Among the fungi rich in polysaccharide content, G. applanatum stands out; however, its anticancer activity necessitates further investigation. This study aims to explore the impact of G. applanatum crude polysaccharide (GACP) extract by assessing its effects on cell viability, levels of proinflammatory cytokines such as TNF-α, IFN-γ, IL-2, and IL-12, and levels of proapoptotic markers including caspase-3 and caspase-9, as well as the percentages of necrosis and apoptosis in the HeLa cell line. Employing the HeLa cell line as a research model, four groups were studied: KN (media and DMSO), K+ (doxorubicin 10 µg/mL), P1 (G. applanatum extract 200 µg/mL), and P2 (G. applanatum extract 400 µg/mL). The G. applanatum extract was obtained via boiling distilled water. Anticancer activity was evaluated through the MTT test (3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide) conducted over three treatment durations (24, 48, and 72 hours). Cytokine levels and caspase-3 and caspase-9 levels were assessed using the ELISA test. Cell apoptosis was determined using the Annexin V-PI biomarker and analyzed through flow cytometry. The MTT test exhibited optimal results at the 48-hour treatment mark. Cytokine level analysis revealed significant reductions in TNF-α, IFN-γ, IL-2, and IL-12 levels (p < 0.005). Concurrently, caspase-3 and caspase-9 levels exhibited substantial increases (p < 0.005). Flow cytometry highlighted the highest percentage of apoptosis in HeLa cells. In conclusion, G. applanatum's polysaccharide extract demonstrates potential as an anticancer and therapeutic agent for cancer treatment.

Effectiveness of ozone-laser photodynamic combination therapy for healing wounds infected with methicillin-resistant Staphylococcus aureus in mice.

Background and Aim: According to 2013 data from the Ministry of Health of the Republic of Indonesia, there were 8.2% more wounds than typical in Indonesia; 25.4% were open wounds, 70.9% were abrasions and bruises, and 23.2% were lacerations. A wound is defined as damage or loss of body tissue. This study aimed to determine the effectiveness of wound healing using red-laser therapy (650 nm, 3.5 J/cm2), blue-laser therapy (405 nm, 3.5 J/cm2), ozone therapy, red-laser therapy (650 nm, 3.5 J/cm2) with ozone, and blue-laser therapy (405 nm, 3.5 J/cm2) with ozone. Materials and Methods: One hundred and twelve mice were given incision wounds and infected with methicillin-resistant Staphylococcus aureus (MRSA). The study used a factorial design with two factors: The type of therapy (n = 7) and irradiation time (days 1, 2, 4, and 6). The mice were divided into seven therapy groups: Control group with NaCl, control with Sofra-tulle® treatment, red-laser therapy (650 nm, 3.5 J/cm2), blue-laser therapy (405 nm, 3.5 J/cm2), ozone therapy, red-laser therapy (650 nm, 3.5 J/cm2) with ozone, and blue-laser therapy (405 nm, 3.5 J/cm2) with ozone. This therapy was performed using irradiation perpendicular to the wound area. The photosensitizer used was curcumin 10 mg/mL, which was applied to the wound area before exposure to a laser and ozone. The ozone concentration was 0.011 mg/L with a flow time of 80 s. The test parameters were the number of collagens, bacterial colonies, lymphocytes, monocytes, and wound length measurement to determine their acceleration effects on wound healing. Data were analyzed by a two-way (factorial) analysis of variance test. Results: Acceleration of wound healing was significantly different between treatments with a laser or a laser-ozone combination and treatment using 95% sodium chloride (NaCl) and Sofra-tulle®. On day 6, the blue-laser with ozone treatment group had efficiently increased the number of bacteria and reduced the wound length, and the red-laser treatment with ozone increased the amount of collagen. In addition, the red-laser also reduced the number of lymphocytes and monocytes, which can have an impact on accelerating wound healing. Blue-laser therapy was very effective for increasing the number of epithelia. Conclusion: The blue- and red-laser combined with ozone treatments effectively accelerated the healing of incisional wounds infected with MRSA bacteria.

In vitro and in vivo antiplasmodial activities of leaf extracts from Sonchus arvensis L.

BACKGROUND: Malaria continues to be a global problem due to the limited efficacy of current drugs and the natural products are a potential source for discovering new antimalarial agents. Therefore, the aims of this study were to investigate phytochemical properties, cytotoxic effect, antioxidant, and antiplasmodial activities of Sonchus arvensis L. leaf extracts both in vitro and in vivo. METHODS: The extracts from S. arvensis L. leaf were prepared by successive maceration with n-hexane, ethyl acetate, and ethanol, and then subjected to quantitative phytochemical analysis using standard methods. The antimalarial activities of crude extracts were tested in vitro against Plasmodium falciparum 3D7 strain while the Peter's 4-day suppressive test model with P. berghei-infected mice was used to evaluate the in vivo antiplasmodial, hepatoprotective, nephroprotective, and immunomodulatory activities. The cytotoxic tests were also carried out using human hepatic cell lines in [3(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] (MTT) assay. RESULT: The n-hexane, ethyl acetate, and ethanolic extracts of S. arvensis L. leaf exhibited good in vitro antiplasmodial activity with IC50 values 5.119 ± 3.27, 2.916 ± 2.34, and 8.026 ± 1.23 µg/mL, respectively. Each of the extracts also exhibited high antioxidant with low cytotoxic effects. Furthermore, the ethyl acetate extract showed in vivo antiplasmodial activity with ED50 = 46.31 ± 9.36 mg/kg body weight, as well as hepatoprotective, nephroprotective, and immunomodulatory activities in mice infected with P. berghei. CONCLUSION: This study highlights the antiplasmodial activities of S. arvensis L. leaf ethyl acetate extract against P. falciparum and P. berghei as well as the antioxidant, nephroprotective, hepatoprotective, and immunomodulatory activities with low toxicity. These results indicate the potential of Sonchus arvensis L. to be developed into a new antimalarial drug candidate. However, the compounds and transmission-blocking strategies for malaria control of S. arvensis L. extracts are essential for further study.

Increasing the effect of annonacin using nanodiamonds to inhibit breast cancer cells growth in rats (Rattus norvegicus)-Induced breast cancer.

Background: Annonaceous acetogenins have been reported to have anti-cancer properties but low viability. In this study, we aimed to investigate the potency of nanodiamonds to be employed as a carrier of annonacin to help increase its viability and inhibit the growth of breast cancer cells. Methods: The annonacin was coupled with nanodiamond and characterized using UV-Vis spectrophotometer, FTIR, SEM, and PSA, and determined their stability and drug release. A cell growth inhibition assay and cell migration assay was performed using the breast cancer MCF7 and T747D cell lines, and in vivo analysis was performed in rats (Rattus norvegicus). MCF7 and T747D cells were treated with 12.5 µg/mL annonacin coupled with nanodiamonds for 24 and 48 h and further analyzed by MTT, cell migration, and reactive oxygen species (ROS) assays. Twenty-five female rats were divided into five groups. Breast cancer was induced using two intraperitoneal doses of N-nitroso-N-methylurea (NMU) (50 and 30 mg/kg body weight). Annonacin coupled with nanodiamonds was administered by intraperitoneal injection (17.5 mg/kg body weight) for 5 weeks, one injection per 3 days. Results: Administration of annonacin coupled with nanodiamonds significantly reduced MCF7 cell growth and reactive oxygen species (ROS) levels. The in vivo study showed that administration of annonacin coupled with nanodiamonds significantly reduced PI3KCA levels and increased p53 expression, reduced cancer antigen-15-3 (CA-15-3) levels in serum, increased caspase-3 expression, reduced Ki-67 levels, and reduced the thickness of the mammary ductal epithelium. Conclusions: Collectively, this study demonstrated the effectiveness of nanodiamonds as a carrier of annonacin to inhibit breast cancer cell growth through inhibition of the PI3K/Akt signaling pathway.

The Regulation of Hypoxia Inducible Factor (HIF)1α Expression by Quercetin: an In Silico Study.

Background: Cancer disease is a growing health problem in developing and developed countries. Hypoxia-inducible factor-1a (HIF1α) is a transcription factor responsible for expressing several proteins involved in angiogenesis. Quercetin can suppress HIF1α expression due to the inhibition of protein synthesis. However, to date, the study exploring the potential of quercetin in repressing HIF1α through its degradation mechanism has never been done. An in silico study is needed as a preliminary study to understand the mechanism underlining this possibility. Objective: This study aimed to investigate the potential of quercetin in regulating HIF1α expression through the ubiquitin degradation pathway by in silico study. Methods: This study was performed by in silico analysis, including biological activity prediction, 3D protein structure collection, protein-ligand and protein-protein docking, and the visualization of the docking results. Results: The probability activity (Pa) score of quercetin as an HIF1α expression inhibitor was 0.969. In the absence of quercetin, the center-weighted score of HIF1α - pVHL, HIF1α - FIH, and HIF1α - PHD2 was -699.4 kJ/mol, -846.0 kJ/mol, and -650.5 kJ/mol, respectively. In the presence of quercetin, the weighted score of HIF1α - pVHL, HIF1α - FIH, and HIF1α - PHD2 was reduced to -728.1 kJ/mol, -854.2 kJ/mol, and -650.5 kJ/mol, respectively. Conclusion: Quercetin could directly promote HIF1α and pVHL interaction, thus increasing the degradation of HIF1α by ubiquitin-dependent pathway.

Effect of crude Ganoderma applanatum polysaccharides as a renoprotective agent against carbon tetrachloride-induced early kidney fibrosis in mice.

Background and Aim: Interstitial fibrosis is the final stage of chronic kidney injury, which begins with an inflammatory process. Crude Ganoderma applanatum polysaccharides are known to have anti-inflammatory properties. The potential role of crude G. applanatum polysaccharides in renal fibrosis through pro-inflammatory cytokines needs further investigation. This study aimed to determine the renoprotective effect of crude G. applanatum polysaccharide extract in mice with carbon tetrachloride (CCL4)-induced early kidney fibrosis. Materials and Methods: This study was conducted for 4 weeks using 24 male BALB/c mice selected for their metabolic stability. The mice were randomly divided into six groups, including control (CG), model (MG), silymarin group and crude G. applanatum polysaccharide extract groups comprising doses of 25, 50, and 100 mg/kg body weight. After sacrificing the mice, whole blood was analyzed for urea and creatine levels, and kidney tissue was prepared to assess tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), hyaluronic acid (HA), and laminin levels, both using enzyme-linked immunosorbent assay. Kidney histology was determined using hematoxylin and eosin staining, while the extracellular matrix (ECM) components were stained using Masson's trichome staining. The α-smooth muscle actin (α-SMA) concentration was determined using immunohistochemistry. These parameters were measured to determine the effectiveness of the crude G. applanatum polysaccharide extract in preventing interstitial fibrosis. Results: Administration of crude G. applanatum polysaccharides effectively prevented increases in kidney weight and physiological enzymes, pro-inflammatory cytokines, and ECM production compared with those in the MG, as evidenced by the low levels of urea, creatinine, TNF-α, IL-6, HA, and laminin. Histopathological results also showed that crude G. applanatum polysaccharides prevented the occurrence of inflammatory infiltration, desquamated nuclei, cytoplasm debris, rupture at the brush border, dilatation of the glomeruli space and lumen of the proximal tubule, and necrotic cells compared with the MG. Masson's trichrome staining revealed lower collagen levels in the interstitial tubules of kidney tissue than those in the MG. Immunohistochemical analysis revealed low α-SMA expression in the crude G. applanatum polysaccharides treatment groups than that in the MG. Conclusion: The crude polysaccharide extract of G. applanatum has a protective effect that prevents the progression of kidney fibrosis in mice.

Anti-cancer activity of an ethanolic extract of red okra pods (Abelmoschus esculentus L. Moench) in rats induced by N-methyl-N-nitrosourea.

Background and Aim: Breast cancer is the most frequent malignancy in women. The consumption of phytochemical components from plants may play an essential role in preventing and treating this cancer. This study aimed to investigate the anti-cancer activity of an ethanolic extract of red okra pods (EEROP) in rats (Rattus norvegicus) induced by N-methyl- N-nitrosourea (MNU). Materials and Methods: The experimental animals were divided into six groups (n=5/group), namely, KN (normal control, without any treatment), K- (negative control, exposed to MNU without EEROP), K+ (positive control, exposed to MNU and Methotrexate), and the treatment Groups P1, P2, and P3 (exposed to MNU and EEROP at doses of 50, 100, and 200 mg/kg body weight [BW], respectively). Intraperitoneal delivery of MNU and EEROP oral administration was carried out for 8 weeks. After the end of treatment, the parameters of cytokines, CD4+ and CD8+ T cells, and mammary gland histology were measured. Results: The results showed that EEROP at doses of 100 and 200 mg/kg BW significantly downregulated interleukin (IL)-6, IL-1ß, tumor necrosis factor (TNF)-α, IL-17, IL-10, and tumor growth factor-ß (p<0.05). In addition, doses of 200 mg/kg BW significantly increased the activity of CD4+ and CD8+ T cells, prevented the proliferation of mammary gland epithelial cells, and yielded a significantly thinner epithelium of the mammary gland (p<0.05). Conclusion: It can be concluded that EEROP was an effective anti-cancer agent by modulating the immune response. Further studies using a nanoparticle system are warranted to achieve optimal working conditions for these bioactive compounds.

Effect of the Ethanol Extract of Red Okra Pods (Abelmoschus esculentus (L.) Moench) to Inhibit Cervical Cancer Cells Growth through Cell Cycle-Associated Oncogenes.

This study aims to evaluate the potency of ethanol extract of red okra pods (EEROP) in inhibiting growth of cervical cancer cells through repression of the cell cycle-associated oncogenes. The EEROP treatment was given to HeLa cells cultured with RPMI medium and incubated at 37°C with 5% CO2. The MTT method was used to measure HeLa cell growth and IC50 values. The mRNA levels of the three cell cycle-associated oncogenes (MYC, TYMS, and MDM2) were evaluated by qRT-PCR to determine the effect of EEROP treatment on the cell cycle. The lowest percentage of viable cells at 24, 48, and 72 hours after EEROP treatment was in the dose of 1000 µg/mL with a growth percentage of 71.60% at 24 hours, 55.61% at 48 hours, and 46.97% at 72 hours. The IC50 values were 2845, 1153, and 776.8 µg/mL for 24, 48, and 72 hours, respectively. The three oncogenes at a dose of 1000 µg/mL significantly decreased the lowest mRNA levels compared to other doses with MYC oncogene that experienced the greatest decrease. The mRNA level of dose 1000 µg/mL EEROP at the MYC oncogene was 0.014-fold changes, at the TYMS oncogene was 0.097-fold changes, and at the MDM2 oncogene was 0.028-fold changes. The EEROP has been shown to decrease the expression of three cell cycle-associated oncogenes. This is also supported by the growth of HeLa cells that did not increase throughout 24, 48, and 72 hours. However, further research is needed on the main active components in red okra that function as anticancer, so that in the future, okra can not only stop cancer cell growth but also induce cancer cell death.

The potential of A. Muricata Bioactive Compounds to Inhibit HIF1α Expression Via Disruption of Tyrosine Kinase Receptor Activity: an In Silico Study.

BACKGROUND: Cancer is a debilitating disease that is on the increase in both developed and developing countries. The plant extract of A. muricata have been known to have a variety of anticancer effects, including anti-angiogenic potential. An in silico study is needed as a preliminary study to understand the mechanism underline this process. OBJECTIVE: The aim of this study was to investigate the potential of the bioactive compounds of A. muricata in regulating angiogenesis process, primarily by the regulation of hypoxia inducible factor (HIF)-1α expression by in silico study. METHODS: This study was performed by in silico analysis including the bioactive compounds preparation, biological activity prediction, protein target and pathway analysis, 3D protein modelling, protein-ligand and protein-protein docking, and the visualization of docking results. RESULTS: There are 3 bioactive compounds of A. muricata with the ability to inhibit HIF-1α expression, including kaempferol, genistein, and glycitein. The inhibition of HIF-1α expression was associated with phosphoinositide 3-kinases (PI3K)/Akt signaling pathway, which involved tyrosine kinase receptor activity on the cell membrane. Based on the silico analysis in this study, we shown that kaempferol, genistein, and glycitein inhibit HIF-1α expression through the disruption of interleukin (IL)-6R and toll-like receptor (TLR)-4 and their respective ligands interaction. CONCLUSION: The findings of this study show that A. muricata bioactive compounds could inhibit HIF-1α expression through disruption of the tyrosine kinase receptor binding with its ligand.

Protective Effect of Red Okra (Abelmoschus esculentus (L.) Moench) Pods against Sodium Nitrite-Induced Liver Injury in Mice.

Vegetables, drinking water, and preserved meats may contain sodium nitrite (NaNO2), which causes liver disease by inducing oxidative stress. Phytochemicals are highly recommended as an alternative to synthetic drugs and affordable medicines to treat liver disease because they have fewer or no side effects. Therefore, this study aims to determine the antioxidant and hepatoprotective potential of red okra fruit ethanol extract against NaNO2-induced liver damage. Thirty-six male mice were separated into six groups. The normal control group (WA) was given distilled water only, and the NaNO2 (SN) group was given only 50 mg/kg BW NaNO2. The other four groups (P1, P2, P3, and P4) were given NaNO2 and red okra ethanol extract at doses of 25, 50, 75, and 100 mg/kg BW, respectively. Gavage was administered orally for 21 consecutive days. Commercial kits define all biochemical parameters according to the manufacturer's instructions. Liver tissue staining followed standard protocols using hematoxylin and eosin. The study revealed that NaNO2 induction causes oxidative stress and damages the liver. The activity of antioxidant enzymes (superoxide dismutase and catalase) significantly increased in the groups treated (P2-P4) with ethanol extract of red okra (p < 0.05). Besides, the oxidants (malondialdehyde, F2-isoprostanes, and nitric oxide) in the liver homogenate significantly decreased in the P4 group, which were given red okra ethanol extract (p < 0.05). Likewise, red okra pods decreased significantly for the serum biochemical parameters of liver damage (aspartate aminotransferase, alkaline phosphatase, and alanine aminotransferase) in the P3 and P4 groups (p < 0.05). Then, it led to a restoration of the histological structure compared to exposed mice (SN), as the pathological scores decreased significantly in the P3 and P4 groups (p < 0.05), as well as the number of the necrotic and swollen liver cells was reduced. Hepatocytes returned to normal. The results showed that the ethanol extract of red okra fruit could be helpful as an affordable medicine. It is an antioxidant and hepatoprotective agent to protect the liver from damage caused by NaNO2.

Dual role of immunomodulation by crude polysaccharide from okra against carcinogenic liver injury in mice.

Polysaccharides have long been recognized as the anticancer agent with low toxicity and slight side effects. Okra (Abelmoschus esculentus L.), a flowering plant from the Malvaceae family that is found in tropical, subtropical, and warm temperate regions around the world. Hence, no in vivo studies have addressed the anticancer and immunomodulatory potential of polysaccharides from okra pods grown in Indonesia. This study aims to investigate the effect of okra raw polysaccharide extract (ORPE) to immune cells and cytokines of mice with hepatocarcinogenic conditions induced by diethylnitrosamine (DEN). Thirty-six male mice (BALB/C, 3-4 months old) were divided into six groups: the normal control group (CN), negative control (C-), positive control giving doxorubicin (C+), and three groups of ORPE treatment given the dose of 50 (P1), 100 (P2) and 200 (P3) mg/kg body weight. The administration of ORPE directly suppressed the regulatory T cells accumulations, suppressed macrophages activations, and balanced the number of effector T cells. However, it promoted CD8+ T cell activation at a low dose and increased interleukin-2 levels at all doses. These results suggest that ORPE has unique dual-functions as immunosuppressant and immunostimulant which can be a foundation for the application of the ORPE in the nutraceutical and pharmaceutical industries.

Hepatoprotective activity of okra (Abelmoschus esculentus L.) in sodium nitrite-induced hepatotoxicity.

BACKGROUND AND AIM: For years, people have used sodium nitrite as a food preservative. This study determined the effect of okra (Abelmoschus esculentus L.) pod methanol extract (OPME) on mice with hepatotoxicity induced by sodium nitrite. The flavonoid and total phenolic levels, serum biochemistry, and liver histology were examined. MATERIALS AND METHODS: Green okra pod extraction was performed using ethanol methanol solvent. Thirty adult male BALB/c mice (8-10 weeks, ~30 g) were divided into six groups: Normal control, negative control (sodium nitrite 50 mg/kg BW exposure), and treatment groups (sodium nitrite exposure and OPME at doses of 50, 100, 200, and 400 mg/kg BW). Subsequently, they were exposed to sodium nitrite and administered multiple doses of OPME for 19 days by gavage. After that, serum was used for biochemical evaluation, and liver histological analysis was performed. All data were statistically analyzed (α=0.05). RESULTS: All doses of OPME reduced the levels of nitric oxide (NO), malondialdehyde (MDA), alanine aminotransferase (ALT), and aspartate aminotransferase (AST). In this research, both superoxide dismutase (SOD) and catalase (CAT) levels increased in all OPME-administered treatments. All doses also reduced necrotic cells, proportion of swollen cells, and inflammation in liver histological analysis. The results of this study showed that OPME exerted hepatoprotective effects by lowering MDA, NO, ALT, and AST levels. It also improved SOD and CAT levels and recovered damaged liver tissue to its normal state. The optimal dose of OPME was 50-100 mg/kg BW. CONCLUSION: OPME has potential as a natural hepatoprotective agent against sodium nitrite exposure.

Antioxidant and Nephroprotective Effects of Okra Pods Extract (Abelmoschus esculentus L.) against Lead Acetate-Induced Toxicity in Mice.

In this study, we determine the curative effects of okra pods (Abelmoschus esculentus L.) extract against lead acetate toxicity in mice kidney. n-Hexane, ethyl acetate, and methanol solvent were used for extracting okra pods. The role of the extract as an antioxidant was tested by DPPH and FRAP methods. The methanol extract was used for experiments in animals. A total of 30 male BALB/c mice were randomly divided into six equal groups: normal control, negative control (lead-induced), and treatment groups (lead-induced for 28 days and administration of methanol extract at doses of 50, 100, 200, and 400 mg/kg BW for the 28 days). The following were analyzed in all groups: activity of the antioxidant enzymes, namely, superoxide dismutase (SOD) and catalase (CAT); oxidant level, namely, malondialdehyde (MDA) and nitric oxide (NO); and markers of kidney injury, namely, blood urea nitrogen (BUN) and creatinine (Cre). Kidney histopathology was also evaluated. This study showed that the methanol extract showed the highest antioxidant activity (IC50 is 35.21 µg/mL, and FRAP is 57.58 µM Fe2+/g). The CAT and SOD activities increased significantly in okra-treated groups (P < 0.05). The okra administration groups experienced a significant decrease in MDA, NO, BUN, and Cre levels (P < 0.05). Thickness of the epithelial proximal tubule, diameter of the proximal tubule, and percentage of necrotic cells in proximal tubule decreased, but the diameter ratio of glomerular Bowman's capsule in mice treated with okra was optimally improved and repaired like normal control (P < 0.05). The results of this study reveal that methanol extract has a very strong antioxidant effect and can reduce the influence of toxicity induced by lead acetate in mice kidney.

Immunomodulatory potential of polysaccharides from Coriolus versicolor against intracellular bacteria Neisseria gonorrhoeae.

BACKGROUND AND AIM: For many years, people use natural products from the plant and fungal to improve immune response against microorganism. This study aimed to investigate the immunomodulatory properties of polysaccharides (PS) from Coriolus versicolor in mice infected by intracellular bacteria Neisseria gonorrhoeae. MATERIALS AND METHODS: Thirty-six female BALB/C mice were divided into six groups: Normal control, negative control, positive control, P1 (PS before infection), P2 (PS after infection), and P3 (PS before and after infection). PS were administrated for 10 days. N. gonorrhoeae was infected twice with 2 weeks gap from the first to second exposure with a dose of 106 cells. 1 week after the end of treatment, level of oxidants, innate immune responses, and adaptive immune responses were measured. RESULTS: This study showed that PS administration could restore the number of leukocytes as normal but could not enhance the number of phagocytes and its activity. PS administration also showed immunosuppression activity by lowering nitric oxide levels in P2 and P3 groups (p<0.05). This result showed that PS prevent over-expression of pro-inflammatory cytokines by decreasing phagocytic activity. Contrast with innate immune response result; PS administration could significantly increase interferon-gamma level in P1, P2, and P3 groups (p<0.05). Level of antibodies was significantly increased in the P3 group (p<0.05). PS administration also showed an increased level of tumor necrosis factor-α, but the difference was not significant (p>0.05). CONCLUSION: PS enhance adaptive immunity due to the capability of N. gonorrhoeae that able to survive and replicate in phagocytes. Thus, PS from C. versicolor could be potentially be used as a natural immunomodulator against intracellular bacteria.

Hepatoprotective effect of crude polysaccharides extracted from Ganoderma lucidum against carbon tetrachloride-induced liver injury in mice.

BACKGROUND AND AIM: Natural products are currently widely used as alternative treatments for liver disease. The study aimed to determine the hepatoprotective effect of crude polysaccharides extracted from Ganoderma lucidum against liver injury induced by carbon tetrachloride (CCl4). MATERIALS AND METHODS: Twenty-four male BALB/C mice were randomly divided into six groups. Serum and liver samples were taken on day 10 after G. lucidum administration. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), malondialdehyde (MDA), superoxide dismutase (SOD), and catalase (CAT) were measured using enzyme-linked immunosorbent assays, and the histology of the liver was evaluated using light microscopy. RESULTS: G. lucidum extract significantly decreased the levels of ALT, AST, and MDA and significantly increased the levels of SOD and CAT. In the histological evaluation, the liver tissue of CCl4-treated mice exhibited hydropic degeneration, necrosis, and sinusoidal dilatation. G. lucidum extract administration improved this liver tissue histopathology. CONCLUSION: Crude polysaccharides extracted from G. lucidum showed a hepatoprotective effect, regenerating damaged liver tissue.

Crude Polysaccharides from Okra Pods (Abelmoschus esculentus) Grown in Indonesia Enhance the Immune Response due to Bacterial Infection.

Okra pods were widely consumed by Indonesians to maintain health. The aim of this study was at investigating the potential of crude polysaccharides from okra pods on immune response in mice infected with Staphylococcus aureus. Thirty male Balb/C mice were divided into six groups: normal control, negative control, and treatment groups (administration of crude polysaccharides at doses of 25, 50, 75, and 100 mg/kg). Crude polysaccharides were administrated for fourteen days. Furthermore, mice were exposed to S. aureus at the fifteenth day. Two weeks after the end of treatment, the parameters were measured. This study showed that crude polysaccharides at a dose of 75 and 100 mg/kg improved phagocytic activity, spleen index, and splenocytes proliferation. Rising of TNF-α levels was shown in groups treated with crude polysaccharides at doses of 25, 50, and 100 mg/kg. All treatment groups showed a decreasing level of IL-17. Crude okra polysaccharides also showed a slight increase in NK cells activity and IFN-γ level. Thus, crude okra polysaccharides could act as an effective material to enhance immune response including phagocytic activity, spleen index, splenocytes proliferation, and control immune responses through cytokine production.