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-1β, tumor necrosis factor (TNF)-α, IL-17, IL-10, and tumor growth factor-β (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.

Background and Aim: Water pollution caused by industrial waste and human activities has disrupted the reproductive health of aquatic organisms. This study aimed to analyze the effects of water pollution caused by polystyrene particles (PP) on the steroid (estradiol and testosterone) levels and histopathology of male tilapia gonads. In addition, we also analyzed the potential of supplementary feeding to remove and neutralize oxidants. Materials and Methods: Thirty-six tilapia fishes were taken for the study and were divided into 12 groups (n=3), including a control group (fed with commercial pellets only) and groups fed with a mixture of commercial-probiotic pellets (200 mL/kg, 1×108 colony-forming unit [CFU]/mL) and commercial vitamin C pellets (100 mg/kg), respectively. The PP concentrations used for this study were 0, 0.1, 1, and 10 mg/L, and the treatment time was 2 weeks. The testosterone and estradiol concentrations were analyzed by enzyme-linked immunosorbent assay and histopathological analysis of the gonads. Results: Laboratory analysis performed using tilapia fishes showed that exposure to a PP concentration of <74 μm, mixed with feed for 14 days, could decrease estradiol and testosterone levels. Exposure to plastic particles could change the structure, shape, and size of male gonads. It can also affect the spermatogenic cell number and alter the diameter inside the cysts. Originally, plastic particles were believed to reduce the permeability of the cyst membrane, and this damages the membrane or ruptures the cyst. Supplementary feed containing probiotics (200 mL/kg, 1×108 CFU/mL) and vitamin C (100 mg/kg) can ameliorate the impact of PP exposure on steroid levels. The steroid levels increase with a concurrent improvement in cysts and seminiferous tubule structures. Conclusion: Overall, this study indicates that PP concentrations in the aquatic environment negatively affect tilapia reproduction, and this may pose a potential threat to the fish population in freshwater. Provision of supplementary feed containing probiotics and vitamin C may serve as an alternative way to counter the negative impact caused by plastic particles.

Background and Aim: Industrial waste, such as heavy metals, is a major source of water pollution; at high levels, such pollution can reduce river water quality to the extent that it becomes unsuitable for aquaculture of freshwater fish. This study aimed to focus on the effects of copper (Cu) exposure in Nile tilapia (Oreochromis niloticus) and specifically the ability of Lactobacillus-based probiotics supplementation in fish feed to ameliorate damage to gonads and negative effects on red blood cells (RBCs), whole blood cells (WBCs), hematocrit (HCT) levels, hemoglobin (HGB) levels, and malondialdehyde (MDA) levels following such exposure. Materials and Methods: Thirty-two Nile tilapia fish were divided into eight groups: A negative control (without probiotics or Cu), a positive control (with probiotics but without Cu), three treatments with probiotics in feed, and one of three Cu concentrations (0.75, 1.50, or 3.00 mg/L), and three treatments with these three Cu concentrations but without probiotics in feed. The probiotics concentration in feed was 25 mL/kg (1×108 CFU/mL). Feeding was for 15 days, after which the hematological parameters, gonadal (testis) structure, and MDA levels of fish were analyzed. Results: Exposure to Cu significantly (p<0.05) affected fish hematology (decreased HGB, HCT, RBC, and WBC levels) and altered the structure of the testes. However, the addition of probiotics to fish feed significantly (p<0.05) ameliorated these effects on hematology and maintained the histological structure of the gonads. Conclusion: Cu exposure at ≥1.5 mg/L affected the hematologic parameters, gonadosomatic index, MDA levels, and testicular cells and tissue of Nile tilapia. However, probiotics supplementation in fish feed helped ameliorate the negative effects of Cu on these parameters. Thus, the Lactobacillus-based probiotics used in this study were apparently able to neutralize Cu toxicity in Nile tilapia.

Polystyrene nanoplastics (PS-NPs) are result from the degradation of plastic and have diameters ranging from 1 nm to 100 nm. The objective of this study is to provide information on the adverse effects of PS-NPs with in vitro and in vivo analyses of liver injury. An in vitro study was conducted using confocal microscopy, flow cytometry, and MTT test analysis. An in vivo study was conducted to determine apoptosis levels, glucose metabolism gene expressions, liver enzymes, and liver histology. Data were analyzed using GraphPad Prism software 10.2.1. The in vitro study showed the absorption of PS-NPs in the cell cytoplasm, the percentage of apoptosis, 3t3, and the WiDr cell lines’ viability. The in vivo analysis showed that PS-NPs can stimulate liver injuries, such as inducing the elevation of liver enzymes, necrosis, edema, inflammation, and the dilatation of the portal vein diameter. High levels of caspase-3, caspase-9, and Bax were detected, as well as the expression of several genes including PI3K, AKT, PEPCK, GLUT2, and PK. In conclusion, the in vitro analysis showed the detrimental effects of PS-NPs on cells, such as high levels of apoptosis and low cell viability, while the in vivo studies displayed the impairment of liver tissue and disturbances in glucose metabolism regulation.

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 (p0.05). 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.

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.