Sida rhombifolia Exerts Anti-Proliferative and Pro-Apoptotic Effects in Human Liver Cancer HepG2 Cells in Vitro.

PURPOSE: Modern research revealed that plants belonging to the Sida rhombifolia family (Malvaceae) contain biologically active compounds that make them prone to discovering and developing anticancer drugs. This study aimed to evaluate the apoptosis effects of S. rhombifolia extracts in HepG2 Cell Line was performed. METHODS: The extractions were prepared, and an MTT assay was applied to evaluate its role in decreasing the viability of HepG2 and HFF cells. Phenolic compounds were analyzed using High-performance liquid chromatography (HPLC). FlowCytometry and RT-qPCR evaluated apoptosis was performed to measure the mRNA expression of pro-and anti-apoptotic mediators. RESULTS: The results can be summarized as EtOAc extract was more cytotoxic against the HepG2 cells (IC50= 364.3 µg/mL) compared to MeOH and HEX extracts (720.2 µg/mL) (560.4 µg/mL) with less cytotoxicity in HFF cells (353.2 µg/mL). The HPLC analysis results revealed most phenolic compounds, such as Epicatechin(1.3 mg/g). The EtOAc extract (300 µg/mL) induced 34% apoptosis in HepG2 cells. RT-qPCR data showed upregulation of the proapoptotic gene (Bax) and increased Bax/BCL-2 ratio by S. rhombifolia EtOAc extract (300 µg/mL). CONCLUSION: In conclusion, the EtOAc extract of S. rhombifolia is capable of inducing apoptosis in HepG2 cells through modulation of the mitochondrial pathway, which explains their antitumor activity.

EISI: Extended inter-spike interval for mental health patients clustering based on mental health services and medications utilisation.

Mental health is vital in all human life stages, and managing mental healthcare service resources is crucial for providers. This paper presents a new method, called Extended Inter-Spike Interval (EISI), on identifying the patients with a similar utilisation of mental health services and medications. The EISI measures the distance between the utilisation patterns of the patients. Then, the pairwise distances are given to a developed split-and-merge Partitioning Around Medoids (PAM) clustering algorithm to identify the patients with similar utilisation patterns. To evaluate the proposed method, we use two years (2013-2014) of the 10% publicly available sample of the Australian Medicare Benefits Schedule (MBS) and Pharmaceutical Benefits Scheme (PBS) administrative data. Results show that mental health patients can be grouped into ten clusters with distinct and interpretable utilisations patterns. The largest cluster comprises individuals who only visit general practitioners and take psycholeptics medications for a short time. The smallest group contains occasional visits with general practitioners and regularly utilises psycholeptics and psychoanaleptics medications over long periods. The proposed method provides insights on whom to target and how to structure services for different groups of individuals with mental health conditions.

Effects of coenzyme Q10 supplementation on inflammation, angiogenesis, and oxidative stress in breast cancer patients: a systematic review and meta-analysis of randomized controlled- trials.

BACKGROUND/OBJECTIVE: Systemic inflammation and oxidative stress (OS) are associated with breast cancer. CoQ10 as an adjuvant treatment with conventional anti-cancer chemotherapy has been demonstrated to help in the inflammatory process and OS. This systematic review and meta-analysis of randomized clinical trials (RCTs) aimed to evaluate the efficacy of CoQ10 supplementation on levels of inflammatory markers, OS parameters, and matrix metalloproteinases/tissue inhibitor of metalloproteinases (MMPs/TIMPs) in patients with breast cancer. METHODS: A systematic literature search was carried out using electronic databases, including PubMed, Web of Science, Scopus, Google Scholar, and Embase, up to December 2020 to identify eligible RCTs evaluating the effect of CoQ10 supplementation on OS biomarkers, inflammatory cytokines, and MMPs/TIMPs. From 827 potential reports, 5 eligible studies consisting of 9 trials were finally included in the current meta-analysis. Quality assessment and heterogeneity tests of the selected trials were performed using the PRISMA checklist protocol and the I2 statistic, respectively. Fixed and random-effects models were assessed based on the heterogeneity tests, and pooled data were determined as the standardized mean difference (SMD) with a 95% confidence interval (CI). RESULTS: Our meta-analysis of the pooled findings for inflammatory biomarkers of OS and MMPs showed that CoQ10 supplementation (100 mg/day for 45-90 days) significantly decreased the levels of VEGF [SMD: - 1.88, 95% CI: (- 2. 62 to - 1.13); I2 = 93.1%, p < 0.001], IL-8 [SMD: - 2.24, 95% CI: (- 2.68 to - 1.8); I2 = 79.6%, p = 0.001], MMP-2 [SMD: - 1.49, 95% CI: (- 1.85 to - 1.14); I2 = 76.3%, p = 0.005] and MMP-9 [SMD: - 1.58, 95% CI: (- 1.97 to - 1.19); I2 = 79.6%, p = 0.002], but no significant difference was observed between CoQ10 supplementation and control group on TNF-α [SMD: - 2.30, 95% CI: (- 2.50 to - 2.11); I2 = 21.8%, p = 0.280], IL-6 [SMD: - 1.56, 95% CI: (- 1.73 to - 1.39); I2 = 0.0%, p = 0.683], IL-1ß [SMD: - 3.34, 95% CI: (- 3.58 to - 3.11); I2 = 0.0%, p = 0.561], catalase (CAT) [SMD: 1.40, 95% CI: (1.15 to 1.65); I2 = 0.0%, p = 0.598], superoxide dismutase (SOD) [SMD: 2.42, 95% CI: (2.12 to 2.71); I2 = 0.0%, p = 0.986], glutathione peroxidase (GPx) [SMD: 2.80, 95% CI: (2.49 to 3.11); I2 = 0.0%, p = 0.543]], glutathione (GSH) [SMD: 4.71, 95% CI: (4.26 to 5.16); I2 = 6.1%, p = 0.302] and thiobarbituric acid reactive substances (TBARS) [SMD: - 3.20, 95% CI: (- 3.53 to - 2.86); I2 = 29.7%, p = 0.233]. CONCLUSION: Overall, the findings showed that CoQ10 supplementation reduced some of the important markers of inflammation and MMPs in patients with breast cancer. However, further studies with controlled trials for other types of cancer are needed to better understand and confirm the effect of CoQ10 on tumor therapy.

Biogenic Silver and Zero-Valent Iron Nanoparticles by Feijoa: Biosynthesis, Characterization, Cytotoxic, Antibacterial and Antioxidant Activities.

BACKGROUND AND PURPOSE: Green nanotechnology is an interesting method for the synthesis of functional nanoparticles. Because of their wide application, they have set up great attention in recent years. OBJECTIVE: The present research examines the green synthesis of Ag and zero-valent iron nanoparticles (AgNPs, ZVINPs) by Feijoa sellowiana fruit extract. In this synthesis, no stabilizers or surfactants were applied. METHODS: Eco-friendly synthesis of Iron and biogenic synthesis of Ag nanoparticles were accomplished by controlling critical parameters such as concentration, incubation period and temperature. Scanning Electron Microscopy (SEM), Transmission Electron Microscope (TEM), Energy-Dispersive X-ray Spectroscopy (EDS), Fourier-Transform Infrared (FT-IR) spectroscopy, X-ray Diffraction analysis (XRD), Dynamic Light Scattering (DLS) and UV-Vis were applied to characterize NPs. The cytotoxicity of NPs was investigated in two cell lines, MCF-7 (breast cancer) and AGS (human gastric carcinoma). A high-performance liquid chromatography (HPLC) analysis was also performed for characterization of phenolic acids in the extract. RESULTS: Both NPs displayed powerful anticancer activities against two tumor cell lines with little effect on BEAS-2B normal cells. Synthesized AgNPs and ZVINPs inhibited the growth of all selected bacteria. Pseudomonas aeruginosa, Proteus mirabilis, Klebsiella pneumonia, Staphylococcus aureus, Enterococcus faecalis, Acinetobacter baumannii and Escherichia coli have been studied in two stages. We initially examined the ATCCs followed by clinical strain isolation. Based on the results from resistant strains, we showed that nanoparticles were superior to conventional antibiotics. DPPH (diphenyl-1-picrylhydrazyl) free radical scavenging assay and iron chelating activity were used for the determination of antioxidant properties. Results showed a high antioxidant activity of scavenging free radicals for ZVINPs and powerful iron-chelating activity for AgNPs. Based on the HPLC data, catechin was the major phenolic compound in the extract. CONCLUSION: Our synthesized nanoparticles displayed potent cytotoxic, antibacterial and antioxidant activities.

Urtica dioica Extract Inhibits Cell Proliferation and Induces Apoptosis in HepG2 and HTC116 as Gastrointestinal Cancer Cell Lines.

BACKGROUND: Nowadays the use of plant-derived products has been extensively examined in the treatment of many types of gastrointestinal cancers such as hepatocarcinoma and colon cancer. Urtica dioica is a traditional herb that has many pharmacological effects and wildly used as a therapeutic agent in cancer. Herein, we have evaluated the effects of the different concentrations of Methanolic Extract of Urtica dioica (MEUD) on viability, death pattern, and expression of the apoptosis-related gene in normal Human Dermal Fibroblast (HDF), hepatocarcinoma cell lines (HepG2) and colon-cancer cell line (HCT116). METHODS: A high-performance liquid chromatography method was developed to simultaneously separate 3 phenolic acids in MEUD. HepG2 and HCT116 cell lines as well as HDF normal cell line were cultured in suitable media. After 24 and 48h, in the cultured cell with different concentrations of MEUD, cells viability was assessed by MTT assay, and apoptosis was also evaluated at the cellular level by Annexin V/PI flow cytometry analyzing and AO/EB staining. BCL2 and BAX gene expressions were assessed by TaqMan real-time PCR assay. RESULTS: MEUD showed antiproliferative effects on HepG2 and HTC116 cells after 48h with an IC50 value of about 410 and 420µg/ml, respectively (P < 0.001). Apoptotic cells were observed in HepG2 and HTC116 cells but not in HDF. Furthermore, the increased level of BAX/BCL-2 ratio was observed in HepG2 and HTC116 cells under the treatment of different concentrations of MEUD. CONCLUSION: The MEUD may influence hepatocarcinoma and colon-cancer cell lines at specific doses and change their proliferation rate by changing the expression of BAX and BCL2.

Highly Concentrated Multifunctional Silver Nanoparticle Fabrication through Green Reduction of Silver Ions in Terms of Mechanics and Therapeutic Potentials.

BACKGROUND: Green synthesis of silver nanoparticles (AgNPs) is limited to produce AgNPs with only relatively low concentrations, and is unsuitable for large-scale productions. The use of Myrtus communis (MC) leaf methanolic extract (rich in hydrolyzable tannins) has been recommended to resolve the issues related to the aggregation of nanoparticles at high concentrations of silver ions with added facet of antioxidant properties. METHODS: The produced highly concentrated MC-AgNPs were characterized by using imaging and spectroscopic methods. Subsequently, antioxidant, anticancer and antifungal activities of the nanoparticles were evaluated. RESULTS: The thermogravimetric analysis and energy dispersive spectroscopy quantitative results suggested that the nanoparticles are biphasic in nature (bio-molecule + Ag0) and layered in structure, suggesting the formation of nanoparticles through a different mechanism than those described in the literature. MC-AgNPs showed greater scavenging activity of nitric oxide and iron (II) chelating ability than the extract. It also showed good reducing power compared to the standard antioxidant. Remarkable anticancer activity of MC-AgNPs (IC50 = 5.99µg/mL) was found against HCT-116 (human colon carcinoma) cell lines after 24h exposure with a therapeutic index value 2-fold higher than the therapeutic index of standard doxorubicin. Furthermore, distinct antifungal activity (MIC = 4µg/mL) was found against Candida krusei. CONCLUSION: The current method outperforms the existing methods because it produces a large amount of multifunctional nanoscale hybrid materials more efficiently using natural sources; thus, it may be used for diverse biomedical applications.

Effect of First Line Gastric Cancer Chemotherapy Regime on the AGS Cell Line - MTT Assay Results.

BACKGROUND: Combination chemotherapy regimes are common treatments for cancer. The aim of this study was to evaluation the effect of individual chemotherapeutic agents in comparison with a first line chemotherapy regime treatment in the AGS gastric cancer cell line by MTT assay. MATERIALS AND METHODS: In this experimental study, AGS cells were grown in RPMI-1640 supplemented with 10% fetal calf serum and 100 IU/ml penicillin, and 10 µg/ml streptomycinin, under a humidified condition at 37° with 5% CO2. All cells were washed with PBS and detached with trypsin, centrifuged and 8000 cells re-plated on to 96- well plates. LD50 doses of Epirubicin, Cisplatin and 5-fluorouracil were added to each well in mono or triple therapy. Anti-proliferative activities were determined by MTT assay after 24, 48 or 72 h. RESULTS: Results of MTT assays showed that there were no significant differences among 3 drugs in monotherapy (p=0.088), but there was significant difference between combination therapy with epirubicin (P=0.031) and 5FU (p=0.013) on cell survival at 24 h. After 48 and 72 hours, cell viability showed significant differences between the 3 drugs (p=0.048 and P=0.000 for 48 and 72 h, respectively) and there was significant difference between combination therapy with epirubicin (P=0.035 and P=0.002 for 48 and 72 h, respectively). CONCLUSIONS: The results showed no significant differences between these chemotherapy drugs each given alone, but combination therapy with 3 drugs had significant effects on cell viability in comparison with epirubicin alone.

Effect of Fibroblast Co-culture on In Vitro Maturation and Fertilization of Mouse Preantral Follicles.

BACKGROUND: The aim of this study was to evaluate fibroblast co-culture on in vitro maturation and fertilization of prepubertal mouse preantral follicles. MATERIALS AND METHODS: The ovaries of 12-14 day old mice were dissected and 120-150 µm intact preantral follicles with one or two layers of granulosa cells, and round oocytes were cultured individually in α-minimal essential medium (α-MEM) supplemented with 5% fetal bovine serum (FBS), 100 mIU/ml recombinant follicle stimulating hormone, 1% insulin, transferrin, selenium mix, 100 µg/ml penicillin and 50 µg/ml streptomycin as base medium for 12 days. A total number of 226 follicules were cultured under two conditions: i) base medium as control group (n=113); ii) base medium co-cultured with mouse embryonic fibroblast (MEF) (n=113). Follicular diameters, alone, in addition to other factors were analyzed by student's t-test and chi-square test, respectively. RESULTS: The co-culture group showed significant differences (p<0.05) in growth rate (days 4, 6 and 8 of the culture period) and survival rate. However, there was no significant difference in antrum formation, ovulation rate and embryonic development of released oocytes. There were significant differences (p<0.05) in the estradiol and progesterone secretion at all days between the co-culture and control groups. CONCLUSION: Fibroblast co-culture increased survival rate and steroid production of preantral follicles by promoting granulosa cell proliferation.