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Curcumin Analogs Induce Apoptosis and G2/M Arrest In 4T1 Murine Triple-Negative Breast Cancer Cells
Chemotherapy is the first-line treatment for triple-negative breast cancer (TNBC), yet toxicity and resistance effects have been the current problems. Curcumin,a natural compound, has been reported to exert anti-proliferative effects on various cancer cells, including breast carcinoma cells. However, the β-diketone moiety influences the stability of curcumin. Curcumin analogs, pentagamavunon-0 (PGV-0), and pentagamavunon-1 (PGV-1) were synthesized to improve the stability and activity of curcumin by modified the β-diketone moiety into mono-ketone pentanone. In this study, we evaluated the cytotoxicity, inhibition of cell cycle progression, and induction of apoptosis of curcumin and its analogs (PGV-0 and PGV-1) in murine triple-negative breast cancer 4T1 cell line. The cytotoxic evaluation was done by MTT assay, while apoptosis induction and cell cycle evaluation was performed by annexin V staining and detected by flow cytometry. Curcumin and its analogs, PGV-0, and PGV-1, significantly inhibit the viability of 4T1 breast cancer cells with an IC50 value of 34.34µg/mL, 13.76µg/mL and 38.21μg/mL, respectively. Apoptosis analysis with a dose of 10µg/mL and 15µg/mL in 4T1 breast cancer cells showed that curcumin and its analogs effectively induce apoptotic in a dose-dependent manner. In cell cycle analysis using a dose of 15µg/mL, curcumin inhibited the cell cycle progression in the S phase, whereas PGV-0 and PGV-1 inhibited the cell cycle in the G2/M phase. It could be concluded that curcumin analogs, PGV-0 and PGV-1, have higher potential to be developed as anti-cancer agents by inducing cell cycle arrest and apoptosis in triple-negative breast cancer
Effects of Peel Extract from Citrus reticulata and Hesperidin, A Citrus Flavonoid, on Macrophage Cell Line
The extract of Citrus reticulata has been studied for its biological activities, due to its citrus flavonoid content. The extract and its flavonoid compounds exhibit growth inhibition properties in several cancer cell lines and in vivo models. Conversely, the extract can also induce cell proliferation and angiogenesis, and shows estrogenic effects, in vitro and in vivo. Because of the contrasting effects that depend on the concentration or dosage, the precise action of the extract and its flavonoids need to be elucidated in various cell types. The objective of this study is to evaluate the effect of Citrus reticulata peel extract (Citrus extract) and hesperidin, a citrus flavonoid, on the modulation of cell proliferation in the RAW 264.7 macrophage cell line. Cell viability under Citrus extract or hesperidin treatment was assessed by using the MTT assay. The expression of interleukin-10 (IL-10), an anti-inflammatory cytokine, modulated by Citrus extract was also examined by immunostaining. Low concentrations of Citrus extract at 1 and 100 μg/mL were able to induce cell proliferation, though not significantly, as shown by cell viability of 138 and 114%, respectively. At higher concentrations of 500, 750, and 1000 μg/mL, Citrus extract decreased cell viability significantly by up to 64, 46, and 36%, respectively. Accordingly, hesperidin at low (3.1 μg/mL−61.1 μg/mL) or high (152.6 μg/mL−305.3 μg/mL) concentrations increased or reduced cell viability significantly by up to 116−136% or 10−61%, respectively. The value of the 50% inhibitory concentration (IC50) of Citrus extract was more than three times higher (756 μg/mL) than that of hesperidin (203 μg/mL = 332 μM). Additionally, 250 μg/mL of Citrus extract was able to induce IL-10 expression compared with the control. These results demonstrate that Citrus extract and hesperidin exert a biphasic effect on macrophage cells. The future development of Citrus extract as a co-chemotherapeutic, anticancer, or immunomodulatory agent should include careful consideration of its biphasic effect on each cell type
In Vitro Study: Effect of Cobalt(II) Chloride Against Dengue Virus Type 1 in Vero Cells
Dengue virus (DENV) serotypes DENV-1 to DENV-4 are enveloped viruses that belong to the genus Flavivirus of the Flaviviridae. Dengue vaccine or antiviral has not yet been clinically approved for humans, even though there have been great efforts toward this end. Antiviral activity against DENV is needed to develop to be an alternative drug for DENV virus. Cobalt(II) chloride have been used in the treatment and prevention of diseases of humans since ancient times. The aim of this study is to investigate the antiviral effects and Cytotoxicity of Cobalt(II) chloride. This compound was further investigated for its inhibitory effect on the replication of DENV-1 in Vero cells. Antiviral activity and Cytotoxicity measured by WST-1 assay. The IC50 value of the Cobalt(II) chloride for DENV-1 was 0.38 μg/ml. The cytotoxicity of Cobalt(II) chloride to Vero cell suggest that the CC50 value was 2.91 µg/ml The results of this study demonstrate the anti-dengue serotype 1 inhibitory activity of Cobalt(II) chloride was a high toxic compound
Potential Deleterious Effects of L-Citrulline Supplementation in Isoproterenol-Induced Myocardial Infarction: Focus on Nitrosative Stress
L-Citrulline shows potential activity as a supplement to prevent myocardial infarction through vasodilative and possible antioxidative effects but may be deleterious by causing nitrosative stress. This study determined the potentially deleterious effects of L-citrulline supplementation in isoproterenol-induced myocardial infarction with a focus on nitrosative stress. L-Citrulline supplementation was given orally at dosages of 300 or 600mg/kg body weight daily for 6 days. Myocardial infarction was induced in Wistar rats via subcutaneous injection of isoproterenol (85 mg/kg body weight (BW)) on day 4 and 5. Blood pressure was measured at the end of the study (day 6) and rats were sacrificed to collect heart tissue samples for a histopathological evaluation. The histopathological evaluation was done using hematoxylin and eosin staining for the myocardial damage evaluation and immunohistochemical (IHC) staining of arginase-2, inducible nitric oxide synthase (iNOS), and 3-nitrotyrosine to evaluate nitrosative stress. L-Citrulline supplementation failed to show a significant protective effect on blood pressure and exacerbated the decrease of diastolic blood pressure. Both low and high dose L-citrulline supplementation had a significant protective effect on myocardial damage compared to the isoproterenol group (p<0.01). L-Citrulline also caused increased nitrosative stress as shown by increased expression of arginase-2 and 3-nitrotyrosine on IHC staining but tended to show an ameliorative effect on iNOS expression. A significant increase in arginase-2 expression was detected between the high dose group and the other groups (p<0.01 vs. normal and isoproterenol groups; p<0.05 vs. low dose group). L-Citrulline supplementation increased 3-nitrotyrosine expression in a dose-dependent manner, which was significantly different compared to the normal group (low dose: p<0.013; high dose: p<0.003). L-Citrulline increased the production of nitrosative stress but resulted in less myocardial damage through its other effects
Cytotoxic Potential of Arthrospira platensis Extract on Cervical Cancer Cells Line Hela: Study on Antiproliferative, Cell Cycle, Apoptosis Induction and Anti Metastasis
Cervical cancer can be treated conventionally with chemotherapy agents, but its use has side effects and complications in the form of damage to normal cells. This study aims to determine the potential of A. platensis as an alternative anticancer agent that is selective towards normal cells. Based on TLC analysis, A. platensis contains antioxidant compounds such as β-carotene, flavonoids, and terpenoids which are able to inhibit proliferation and trigger apoptosis of cancer cells. The study was conducted using cervical cancer cells HeLa and normal cells HDFa. A. platensis macerated with 96% ethanol at a ratio of 1:4. Based on probit analysis, it is known that ethanol extract of A. platensis has a cytotoxic effect on HeLa cells with IC50 values of 260.444μg/mL and index selectivity towards HDFa cells of 7.931. The mechanism of cytotoxic activity of ethanol extract of A. platensis is related to its ability to extend the doubling time, increase the induction of apoptosis, and reduce the rate of cells migration. Ethanol extract of A. platensis can also increase cells accumulation in the S phase to prevent cells from entering the G2/M phase
Effect of Growth Factor In Callus Induction and Bioactive Compounds In Seed Explant of Kaffir Lime (Citrus hystrix DC.)
Our previous study showed that kaffir lime leaf extracts may have anti-cancer properties. However, production of bioactive compounds is affected by environmental factors. Here, we present a method to control environmental conditions using in vitro culture techniques such as callus induction. Calluses were induced from seed embryo explants of kaffir lime on MS medium with combinations of 2,4-D and BAP at concentrations 1:0.5; 1:1; and 2:1, respectively. Fourty and 60 days-old calluses were extracted using chloroform and ethyl acetate and analyzed by GC-MS. Results showed all combinations of 2,4-D and BAP were able to induce callogenesis from seed embryo explants of kaffir lime with no significant differences of callus initiation time, biomass, morphology and growth rates. However differences were detected in the bioactive compound profiles. In kaffir lime callus, both fatty acids and secondary metabolites were detected. Specifically, in 40 days-old calluses (exponential growth phase) we detected α-pinene and 1.8–cineole in plants treated with 2,4-D: BAP at concentration 1:0.5 and 2:1. In 60 days-old calluses (stationary phase) we detected a number of compounds in plants treated with 2,4-D:BAP at concentrations of 1:0.5 and 2:1, including caryophyllene, linoleoyl chloride, thiogeraniol, stigmasterol, clianosterol, citronellal, neo-isopulegol, citronellol, geraniol, eugenol, cyclopropane, pristane, elemol and farneso
QuEChERS and C18 as solid phase extraction sorbent - ultra-high performance liquid chromatography -ultraviolet-visible method for determination of parabens in cosmetics products
Concerns are growing about human exposure to endocrine-disrupting chemicals (EDCs), especially during the preadolescent development stage. Parabens are prevalent EDCs widely used as additives in cosmetics. So, the determination of parabens in such products is important. In this study, we developed a reliable and sensitive method to determine simultaneously nine common parabens (methylparaben, ethylparaben, phenylparaben, benzylparaben, penthylparaben, and two groups of isomeric compounds include propylparaben, isopropyl paraben, and butylparaben, isobutylparaben) in cosmetics products. The QuEChERS and solid-phase extraction techniques are used for extraction parabens from non-surfactant cosmetics (perfume, mouth wash solution) and surfactant cosmetics (shampoo, cream, gel), respectively and quantified by using ultra-performance liquid chromatography coupled with the ultraviolet-visible detector. All nine compounds showed good linearity with regression coefficients predominantly above 0.990. The LOD and LOQ of parabens were 0.07 µg/mL; 0.2 µg/mL, respectively. The recoveries ranged from 80 to 110% with the relative standard deviations below 8%. The developed method was successfully applied to determine parabens in various commercial cosmetic products from a local supermarket and the total parabens concentrations are in a wide-ranged from 2.0 to 1270 mg/kg
The effect of combination of active fraction Andrographis paniculata (Burm.f) Ness and Centella asiatica (i) Urban on the alpha glucocidase inhibitor and antioxidant activities
The Andrographis panicullata and Centella asistica extract have been reported that had a anti-diabetic effect. However, the specific mechanism and the effect combination of both were not yet reported. This study was purposed to determine the potency of extract, fractions and the combination of Andrographis panicullata (AP) and Centella asistica (CA) active fraction to inhibit alpha glucosidase enzymeand its ability to reduce DPPH radical.AP and CA were extracted using 50% ethanol then fractionated with solvents under different polarity levels. The inhibiting activity to alpha glucosidase enzyme and antioxidant activity of each fractions was tested. The most active fractions from AP and CA were then combined and re-tested for activity. The results result reported that both of AP and CA had inhibition of alpha glucosidase activity and antioxidant activity. Based on calculation combination index (CI) of of active fraction of AP and CA showing in alpha glucocidase activity had a antagonist action and antioxidant had a sinergic action. Therefore, combination of AP and CA not has not recomended for alpha glucocidase inhibitor but the combination has ability to reduce DPPH radical
In Vitro Antiplasmodial Activity and Cytotoxicity of Active Subfractions of Harmsiopanax aculeatus Leaves
Harmsiopanax aculeatus leaves, a medicinal plant with locally named kapur, have been used traditionally to treat malaria in Maluku, Indonesia. However, the scientific information of this plant is still limited. In our previous study, the methanol extract of this plant leaves have been proven to possess in vitro antiplasmodial activity. This study was conducted to evaluate in vitro antiplasmodial activity and cytotoxicity of subfractions of the plant leaves. Fractionation was performed using a column chromatography with Sephadex LH-20 as the stationary phase and methanol as the mobile phase. The subfractions obtained were then tested for in vitro antiplasmodial activity on a chloroquine-resistant FCR3 strain of Plasmodium falciparum using a visual method. Cytotoxicity was evaluated by using MTT assay. The in vitro antiplasmodial activity and cytotoxicity were expressed as IC50, calculated using probit analysis with SPSS 16 for windows. The results showed that the four subfractions tested have a high antiplasmodial activity with IC50 values of 0.09; 0.18; 0.01; and 0.77 µg.mL-1, respectively. In addition, these subfractions had IC50 values of >400 µg.mL-1 against Vero cells indicating that they were non-toxic. In conclusion, the subfractions of H. aculeatus leaves are very active and selective against P. falciparum. Further study will be conducted to isolate the active compounds
FTIR Method for Peptide Content Estimation and Degradation Kinetic Study of Canarium Nut Protein
Quantitative analysis of bioactive peptide mostly conducted by measuring the activity. While the determination of peptide content in natural sources has been conducted using various instruments, vibrational spectroscopy remains underutilized. Here, we attempted new developed method in peptide quantification and degradation kinetic analysis using Fourier-Transform Infrared Spectroscopy. Bovine Serum Albumin was used as standard protein in method development and validation. Peptide content was estimated by converting peak area to concentration. The method was used to estimate peptide content in Canarium nut protein and its hydrolysates, which potentially hold biological activity. Kinetic study was conducted with microwave as an accelerator for hydrolysis, an apparatus rarely used in peptide study. Amide I band on wavenumber range of 1724.05-1619.91 cm-1 was selected for analysis, considering its selectivity and linearity. The method also met other validation requirement, including accuracy and precision. When applied in quantitative analysis, the method was able to calculate peptide content decrease in Canarium nut protein after hydrolysis using papain (38.24%), pepsin (33.67%) and alkaline reagent (28.53%). In kinetic study, microwave-assisted peptide degradation exhibited logarithmic profile with the equation of y=-0.148ln(x)+0.9591 and R² value of 0.963. Based on these results, FTIR is useful in estimating peptide content and in analyzing degradation kinetic profile.