Correlation of High-Grade Osteosarcoma Response to Chemotherapy with Enhanced Tissue Immunological Response: Analysis of CD95R, IFN-γ, Catalase, Hsp70, and VEGF.

High-grade osteosarcoma, a primary malignant bone tumour, is experiencing a global increase in reported incidence with varied prevalence. Despite advances in management, which include surgery and neoadjuvant chemotherapy often an unsatisfactory outcome is found due to poor or heterogeneous response to chemotherapy. Our study delved into chemotherapy responses in osteosarcoma patients and associated molecular expressions, focusing on CD95 receptor (CD95R), interferon (IFN)-γ, catalase, heat-shock protein (Hsp)70, and vascular endothelial growth factor (VEGF). Employing immunohistochemistry and Huvos grading of post-chemo specimens, we analysed formalin-fixed paraffin-embedded (FFPE) osteosarcoma tissue of resected post-chemotherapy specimens from Dr. Soetomo General Academic Hospital in Surabaya, Indonesia (DSGAH), spanning from 2016 to 2020. Results revealed varied responses (poor 40.38%, moderate 48.08%, good 11.54%) and distinct patterns in CD95R, IFN-γ, catalase, Hsp70, and VEGF expression. Significant differences among response groups were observed in CD95R and IFN-γ expression in tumour-infiltrating lymphocytes. The trend of diminishing CD95R expression from poor to good responses, accompanied by an increase in IFN-γ, implied a reduction in the count of viable osteosarcoma cells with the progression of Huvos grading. Catalase expression in osteosarcoma cells was consistently elevated in the poor response group, while Hsp70 expression was highest. VEGF expression in macrophages was significantly higher in the good response group. In conclusion, this study enhances our understanding of immune-chemotherapy interactions in osteosarcoma and identifies potential biomarkers for targeted interventions.

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.

The effect of laparoscopy on mast cell degranulation and mesothelium thickness in rats.

BACKGROUND: Laparoscopy induces adhesion due to ischemia-reperfusion injury. However, the detail pathomechanism is poorly understood. This study aimed to investigate the impact of laparoscopy on mast cell and mesothelium morphological changes in the rat. METHODS: Forty-nine males of Sprague-Dawley Rattus norvegicus were divided into four groups: a) control and b) intervention groups P1, P2, and P3 that underwent 60 min laparoscopic using carbon dioxide (CO2) insufflation at 8, 10, and 12 mmHg groups, respectively. Serum hydrogen peroxide (H2O2), catalase (CAT), superoxide dismutase (SOD), malondialdehyde (MDA), and oxidative stress index (OSI) levels were determined 24 h after laparoscopy. Histopathological analyses of mast cell infiltration and degranulation and mesothelium thickness in the liver, greater omentum, mesenterium, small intestine, and peritoneum were performed 7 days after the procedure. RESULTS: H2O2, MDA, and OSI levels were significantly increased in the intervention groups compared with the control (p<0.05), while the SOD and CAT levels were decreased in the intervention groups compared with the control (p<0.05). Mast cell infiltration and degranulation were higher in the intervention groups than in control (p<0.05), while the mesothelium thickness was significantly lower in the laparoscopic groups than in control (p<0.05). Interestingly, the decrease in mesothelium thickness was strongly associated with the increase in mast cell infiltration and degranulation (p<0.01). CONCLUSIONS: Our study shows that laparoscopy in rats increases mast cell infiltration and degranulation, which also results in and correlates with a decrease in mesothelial thickness.

Interleukin 10 Induces the Expression of Membrane-Bound HLA- G and the Production of Soluble HLA-G on HeLa CCL-2 Cells.

BACKGROUND: Interleukin-10 is a cytokine that has a pleiotropic effect on the immune system and inflammation. IL-10 can contribute to the anti-tumour immune response by increasing HLA-G expression. AIM: This study aimed to determine the effect of IL-10 induction on membrane HLA-G expression and soluble HLA-G production of HeLa CCL-2 cells. METHODS: HeLa CCL-2 cells were cultured in the well plate and divided into 4 groups consist of 1 control group that was not induced by IL-10 and 3 treatment groups that were induced by IL-10 500 ng/ml, 1000 ng/ml and 2000 ng/ml respectively. All groups were incubated for 48 hours in a 37°C incubator at 5% CO2 atmospheric pressure. HLA-G measurements were carried out both in cell lysate and cell culture supernatant using ELISA and in membrane-bound using immunofluorescence method. The expression of HLA-G in membrane-bound calculated using the ImageJ application. Data obtained were analysed by ANOVA and LSD test. RESULTS: In the control group, the HLA-G level in the culture supernatant was higher than in cell lysate (p = 0.000), as well as in all treatment groups (p = 0.000). There were significant differences between the treatment group (p = 0.000) and within the treatment group (p = 0.000) at HLA levels. The highest expression of HLA-G in HeLa cell membranes found in cell culture induced by IL-10 concentrations of 500 ng/ml, i.e., 59.28 AU in view. HLA-G membrane expression in the IL-10 1000 ng/ml induced group was significantly different from all treatment groups (p = 0.000). CONCLUSION: HeLa CCL-2 cells express HLA-G on the membrane and release dissolved HLA-G without induction of IL-10 although IL-10 induction augments the presence and the production of HLA-G in HeLa CCl-2 cells.

Oxidants-Antioxidants Profile in the Breast Cancer Cell Line MCF-7

Reactive oxygen species (ROS) have various biological effects and they are non-linear in characteristic. In high oxidative stress, they may cause cytotoxicity, inhibit cell proliferation, and induce cell death in the form of apoptosis/necrosis; while in low or medium oxidative stress, ROS may cause DNA damage, cell mutation, inflammation, cell proliferation, and eventually they may induce carcinogenesis. Antioxidants are compounds with the ability to reduce ROS. Cell line MCF-7 is one of the breast cancer cell lines that is known to have small amount of antioxidant MnSOD compared to the other cell lines. Low antioxidant MnSOD level in breast cancer cell line MCF-7 leads to low concentration of hydrogen peroxide, because antioxidant MnSOD will convert radical superoxide to hydrogen peroxide. The aim of this research was to analyze oxidants and antioxidants profile in breast cancer cell line MCF-7 and their relationship with cell number. Observations were conducted for 5 days. The cell number was counted with tryphan blue method and haematometer. The concentration of radical superoxide was measured with DHE staining using LSCM tipe Olympus Fluoview FV 1000-Ver 1.7. MnSOD activity, hydrogen peroxide concentration, and catalase activity were measured with ELISA. The results showed that the longer of observation, the greater concentration of oxidants and MnSOD activity, but there was no change in catalase activity. Conclusion the increase in cancer cells number is influenced by radical superoxide.

Strong renal expression of heat shock protein 70, high mobility group box 1, inducible nitric oxide synthase, and nitrotyrosine in mice model of severe malaria.

INTRODUCTION: Renal damage is a consequence of severe malaria, and is generally caused by sequestration of Plasmodium falciparum -infected erythrocytes in the renal microcirculation, which leads to obstruction, hypoxia, and ischemia. This triggers high mobility group box 1 (HMGB1) to send a danger signal through toll-like receptors 2 and 4. This signal up-regulates inducible nitric oxide (iNOS) and nitrotyrosine to re-perfuse the tissue, and also increases heat shock protein 70 (HSP70) expression. As no study has examined the involvement of intracellular secondary molecules in this setting, the present study compared the renal expressions of HSP70, HMGB1, iNOS, and nitrotyrosine between mice suffered from severe malaria and normal mice. METHODS: C57BL/6 mice were divided into an infected group (intraperitoneal injection of 10 6 P. berghei ANKA) and a non-infected group. Renal damage was evaluated using hematoxylin eosin staining, and immunohistochemistry was used to evaluate the expressions of HSP70, HMGB1, iNOS, and nitrotyrosine. RESULTS: Significant inter-group differences were observed in the renal expressions of HSP70, HMGB1, and iNOS (p=0.000, Mann-Whitney test), as well as nitrotyrosine (p=0.000, independent t test). The expressions of HSP70 and HMGB1 were strongly correlated (p=0.000, R=1.000). No correlations were observed between iNOS and HMGB, HMGB1 and nitrotyrosine, HSP70 and nitrotyrosine, or iNOS and nitrotyrosine. CONCLUSIONS: It appears that HMGB1, HSP70, iNOS, and nitrotyrosine play roles in the renal damage that is observed in mice with severe malaria. Only HSP70 expression is strongly correlated with the expression of HMGB1.

Strong renal expression of heat shock protein 70, high mobility group box 1, inducible nitric oxide synthase, and nitrotyrosine in mice model of severe malaria

Abstract INTRODUCTION Renal damage is a consequence of severe malaria, and is generally caused by sequestration of Plasmodium falciparum -infected erythrocytes in the renal microcirculation, which leads to obstruction, hypoxia, and ischemia. This triggers high mobility group box 1 (HMGB1) to send a danger signal through toll-like receptors 2 and 4. This signal up-regulates inducible nitric oxide (iNOS) and nitrotyrosine to re-perfuse the tissue, and also increases heat shock protein 70 (HSP70) expression. As no study has examined the involvement of intracellular secondary molecules in this setting, the present study compared the renal expressions of HSP70, HMGB1, iNOS, and nitrotyrosine between mice suffered from severe malaria and normal mice. METHODS C57BL/6 mice were divided into an infected group (intraperitoneal injection of 10 6 P. berghei ANKA) and a non-infected group. Renal damage was evaluated using hematoxylin eosin staining, and immunohistochemistry was used to evaluate the expressions of HSP70, HMGB1, iNOS, and nitrotyrosine. RESULTS Significant inter-group differences were observed in the renal expressions of HSP70, HMGB1, and iNOS (p=0.000, Mann-Whitney test), as well as nitrotyrosine (p=0.000, independent t test). The expressions of HSP70 and HMGB1 were strongly correlated (p=0.000, R=1.000). No correlations were observed between iNOS and HMGB, HMGB1 and nitrotyrosine, HSP70 and nitrotyrosine, or iNOS and nitrotyrosine. CONCLUSIONS It appears that HMGB1, HSP70, iNOS, and nitrotyrosine play roles in the renal damage that is observed in mice with severe malaria. Only HSP70 expression is strongly correlated with the expression of HMGB1.