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.

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.

Topical Administration Effect of Sargassum duplicatum and Garcinia mangostana Extracts Combination on Open Wound Healing Process in Diabetic Mice.

This research aimed to determine the topical administration effect of the combination of Sargassum duplicatum and Garcinia mangostana extracts to ameliorate diabetic open wound healing. The study used 24 adult males of Mus musculus (BALB/c strain, 3-4 months, 30-40 g). They were divided into normal control groups (KN) and diabetic groups. The diabetic group was streptozotocin-induced and divided further into three treatment groups: the diabetic control group (KD), the S. duplicatum treatment group (PA), and the combination of S. duplicatum and G. mangostana treatment group (PAM). The dose of treatment was 50 mg/kg of body weight. Each group was divided into three treatment durations, which were 3 days, 7 days, and 14 days. The wound healing process was determined by wound width, the number of neutrophils, macrophages, fibroblasts, fibrocytes, and collagen density. Histological observation showed that the topical administration of combination extracts increased the re-epithelialization of the wounded area, fibroblasts, fibrocytes, and collagen synthesis. The topical administration of combination extracts also decreased the number of neutrophils and macrophages. This study concluded that the topical administration of the combination of S. duplicatum and G. mangostana extracts improved the open wound healing process in diabetic mice.

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.

The combined effect of magnetic and electric fields using on/off infrared light on the blood sugar level and the diameter of Langerhans islets of diabetic mice.

BACKGROUND AND AIM: At present, diabetes is treated with oral antidiabetic medicines, such as sulfonylureas and thiazolidine, as well as insulin injection. However, these methods have several shortcomings. Therefore, alternatives for treating diabetes mellitus (DM) are needed. This study aims to determine the combined effect of magnetic and electric fields on blood sugar levels and the diameter of Langerhans islets of diabetic mice. MATERIALS AND METHODS: Induction of DM in mice was carried out by administering lard for 2 weeks and continued with an intraperitoneal injection of streptozotocin, dissolved in a 4.5 pH citrate buffer, and administered in a dose of 30 mg/kg bodyweight for 5 days. Treatments were used in combination with magnetic and electric fields using on/off infrared light. Blood samples were pipetted through the tip of mice's tails to establish the blood sugar level for each individual mouse. Histology preparation of the pancreas organ was affected using the histology standard as well as hematoxylin and eosin staining methods. Langerhans islet diameter data were analyzed using analysis of variance followed by Duncan's multiple range test. Data analysis was performed at ssssssss=0.05. RESULTS: The results showed that the combined treatment of permanent magnetic and unidirectional electric fields (PS) caused changes in blood sugar levels that were not significantly different from the normal control group. The PS treatment improved the diameter of the Langerhans islets but not to a significant degree compared to other treatments. CONCLUSION: The use of PS treatment is effective for reducing the blood sugar levels of diabetic mice and improving the diameter of their Langerhans islets.

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.

Efficacy and Tolerability of Topical Nicotinamide Plus Antibacterial Adhesive Agents and Zinc-Pyrrolidone Carboxylic Acid Versus Placebo as an Adjuvant Treatment for Moderate Acne Vulgaris in Indonesia: A Multicenter, Double-blind, Randomized, Controlled Trial.

OBJECTIVE: We investigated the efficacy and tolerability of nicotinamide cream plus an antibacterial adhesive agent and zinc-pyrrolidone carboxylic acid compared to placebo in patients with moderate acne vulgaris (MAV) in Indonesia. METHODS: This was a multicenter, double-blind, randomized, placebo-controlled, parallel-group study conducted in five teaching hospitals in Indonesia from August 2016 to January 2017. Eligible participants included 140 patients with MAV, aged 12 to 50 years, who were enrolled and randomly divided into two groups to receive either adapalene and the study formulation or adapalene and a placebo cream twice daily for six weeks. Clinical response and treatment efficacy were assessed through acne lesion counts, presence of side effects, and patient satisfaction at the second, fourth, and sixth weeks after the first visit. RESULTS: A total of 140 subjects from five different centers (28 subjects in each center) were enrolled. One hundred twenty-seven subjects completed the study, including 63 subjects in the study group and 64 subjects in the placebo group. A significant decrease in the number of noninflammatory lesions in the second week was noted in the study group compared to in the placebo group. There were no significant differences in adverse effects between the two groups in the second and fourth weeks. CONCLUSION: Treatment using nicotinamide plus an antibacterial adhesive agent and zinc-pyrrolidone carboxylic acid was effective in reducing noninflammatory lesions by the second week of therapy. ClinicalTrials.gov registration no. NCT0326298.

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.

The effect of electric field, magnetic field, and infrared ray combination to reduce HOMA-IR index and GLUT 4 in diabetic model of Mus musculus.

Diabetes mellitus (DM) is a metabolic disorder characterized by high blood glucose level (hyperglycemia). Type 2 diabetes mellitus is mainly featured by low cell sensitivity towards insulin stimulation, caused by ectopic fat storage. Insulin resistance can be quantified from high number of HOMA-IR index and observed from glucose transporter 4 (GLUT-4) translocation on membrane of skeletal muscle cells. Combined treatment of electric field, magnetic field, and infrared ray have potential to reduce insulin resistance due to improving blood circulation and increasing intracellular Ca2+ level. The aim of study was to determine the effect of electric field, magnetic field, and infrared ray combination to lower insulin resistance in the type II diabetic model of Mus musculus. This study used 30 adult male mice strain BALB/c. Diabetes was induced using high-fat diet/streptozotocin method until random blood glucose level reached > 200 mg/dL. Diabetic mice were then exposed to electrical field (static and dynamic), magnetic field (static and induce), and infrared ray (with or without infrared ray) combination therapy 15 min daily for 28 days. Fasting blood glucose level, plasma insulin level, HOMA-IR index, and membrane GLUT-4 density after treatment were analyzed statistically at α = 0.05. Result showed that exposure combination of electrical field, magnetic field, and infrared were found to be able to lower fasting blood glucose level and HOMA-IR index significantly, but plasma insulin level and GLUT-4 density were not found to be significantly different compared to diabetic control. Based on current study result, the best combination for reducing insulin resistance in diabetic mice is BsEsI (combination of static magnetic field (Bs), static electric field (Es), with infrared (I)), indicated by lowest HOMA-IR compared to other groups. Exposure to combination of magnetic field, electrical field, and infrared resulted in lowering fasting blood glucose level and HOMA-IR index in diabetic mice, indicating reduced insulin resistance.

The effects of laser diode treatment on liver dysfunction of Mus musculus due to carbofuran exposure: An in vivo study.

OBJECTIVE: The aim of this study is to determine the effect of laser diode as an alternative treatment on liver dysfunction (in vivo study) that is caused by carbofuran using male mice (Mus musculus) strain Balb/C. MATERIALS AND METHODS: The samples were divided into three groups, namely, Group C-L- (control group, no treatment), Group C+L- (only treated by carbofuran treatment), and Group C+L+ (treatment group, treated by carbofuran and laser-puncture) with five replications each. After being treated, each liver slice of samples was observed using microscope to get the histology result and then scored. RESULTS: Carbofuran contamination can lead to inflammation of cells and necrosis. The histology results and the scoring test showed that the liver cells repair with the energy dose of laser diode at 0.5 and 1.0 Joule. CONCLUSION: The optimum energy dose in this study was 1.0 Joule which had the closest score of inflammatory cells and necrosis to normal liver cells.

In vivo approach on femur bone regeneration of white rat (Rattus norvegicus) with the use of hydroxyapatite from cuttlefish bone (Sepia spp.) as bone filler.

BACKGROUND: Hydroxyapatite (HA) from bovine bone has been widely used as bone filler in many fractures cases. HA can also be made from cuttlefish bone (Sepia spp.) that has abundant availability in Indonesia and contains 84% CaCO3, which is a basic ingredient of HA. However, research on the effects of HA from cuttlefish bone on bone regeneration parameters has not been done yet. AIM: This study aimed to determine femur bone regeneration of white rats (Rattus norvegicus) through the use of HA from cuttlefish bone (Sepia spp.) as bone filler. MATERIALS AND METHODS: HA was made using the hydrothermal method by mixing 1M aragonite (CaCO3) from cuttlefish bone and 0.6 M NH4H2PO4 at 200°C for 12 h followed by sintering at 900°C for 1 h. In vivo test was carried out in three groups, including control group, bovine bone-derived HA group, and cuttlefish bone-derived HA group. The generation of femur bone was observed through the number of osteoblasts, osteoclasts, woven bone, lamellar bone, havers system, and repair bone through anatomical pathology test for 28 days and 56 days. RESULTS: Anatomical pathology test results are showed that administration of bovine bone-derived HA and cuttlefish bone-derived HA increased the number of osteoblasts, osteoclasts, woven bone, lamellar bone, havers system, and bone repair at recuperation of 56 days. Statistical test using Statistical Package for the Social Sciences with Kruskal-Wallis and Mann-Whitney U-test was resulted in significant differences between the bovine bone-derived HA control group and the cuttlefish-derived HA control group. There was no significant difference toward the indication of bone formation through the growth of osteoblasts, osteoclasts, woven bone, lamellar bone, havers system, and bone repair in the bovine bone-derived HA and cuttlefish bone-derived HA groups. CONCLUSION: It can be concluded that cuttlefish bone-derived HA has the potential as bone filler based on the characteristics of bone regeneration through in vivo test.

Anticancer Activity of Methanol Extract of Ficus carica Leaves and Fruits Against Proliferation, Apoptosis, and Necrosis in Huh7it Cells.

The polyphenol plant extracts have previously been demonstrated to act as chemopreventive and anticancer agents. Ficus carica is a rich source of polyphenols, yet its antioxidant and anticancer activities remain poorly characterized. This study aimed to determine the anticancer activity of F carica leaf and fruit extracts by investigating their impact on proliferation, apoptosis, and Huh7it cell necrosis. Leaves and fruits were extracted using methanol, and the phytochemical contents were analyzed using Fourier-transform infrared spectroscopy. The antioxidant activity was measured using the 2,2-diphenyl-1-picrylhydrazyl method. Anticancer activities were examined through MTT (3-(4,5-dimethylthiazol-2yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) assay on Huh7it liver cancer cells. The apoptosis and necrosis conditions were examined using Annexin biomarkers V-PI and later analyzed in flow cytometry. F carica leaves and fruit examined were found to have strong antioxidant activities with IC50 values of 7.9875 µg/mL and 13.402 µg/mL, respectively. MTT assay results indicated F carica leaves and fruit had IC50 values >653 µg/mL and >2000 µg/mL, respectively. The flow cytometry analysis indicated a higher percentage of Huh7it apoptosis and necrosis in leaf extracts compared with fruit extracts. The difference in anticancer activity was attributed to differing compounds present in each extract.

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.

ACANTHAMOEBA SP.S-11 PHAGOCYTOTIC ACTIVITY ON MYCOBACTERIUM LEPRAE IN DIFFERENT NUTRIENT CONDITIONS.

BACKGROUND: Mycobacterium leprae (M. leprae) is a pathogenic bacterium that causes leprosy. The presence of M. leprae in the environment is supported by microorganisms that act as the new host for M. leprae. Acanthamoeba's potential to be a host of M. leprae in the environment. Acanthamoeba sp. is Free Living Amoeba (FLA) that classified as holozoic, saprophytic, and saprozoic. The existence of nutrients in the environment influence Acanthamoeba ability to phagocytosis or pinocytosis. This study is aimed to determine Acanthamoeba sp.S-11 phagocytic activity to Mycobacterium leprae (M. leprae) which cultured in non-nutrient media and riched-nutrient media. MATERIALS AND METHODS: This research conducted by culturing Acanthamoeba sp.S-11 and M. leprae on different nutrient media conditions. M. leprae intracellular DNA were isolated and amplified by M. leprae specific primers through Real Time PCR (Q-PCR). RESULT: The results showed that Acanthamoeba co-cultured on non-nutrient media were more active to phagocyte M. leprae than on rich-nutrient media. CONCLUSION: The use of non-nutrient media is recommended to optimize Acanthamoeba sp. phagocytic activity to M. leprae.