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Eugenol is a phenolic compound isolated from clove essential oil. It is used in dentistry, fragrance, cosmetic, and food industries. According to the fragrance ingredient safety assessment report, eugenol does not cause phototoxic reactions and genotoxicity. However, its effect on UV-induced cytotoxicity and genotoxicity has not been well examined. Here in this study, cytotoxic and genotoxic effects of eugenol are investigated on UVA-induced damage using human keratinocyte cells (HaCaT). HaCaT cells were treated with increasing concentrations of eugenol (10-500 μM) for 1 hour and irradiated with 5-10-15 J/cm2 UVA. 24 hours later the neutral red uptake (NRU) assay was used to evaluate cytotoxicity. For genotoxicity assay cells were exposed to 1-10 μM eugenol for one hour and non-cytotoxic UVA irradiation doses (1, 2.5 J/cm2) were used. The alkaline comet assay was carried out immediately after the UVA irradiation to measure the genotoxic potential of eugenol. The cytotoxicity assay results indicate that eugenol caused a cytotoxic effect in a dose-dependent manner in HaCaT cells and increasing doses of UVA-irradiation enhanced the cytotoxic effect of eugenol. The alkaline comet assay results showed that eugenol causes DNA single-strand breaks and increasing doses of UVA-irradiation aggravates the genotoxic potential of eugenol. These data demonstrate that eugenol has cytotoxic and genotoxic potential and eugenol aggravates UVA-induced cytotoxic and genotoxic response in HaCaT human keratinocytes.

BackgroundOur production unit realises more than 35 000 cytotoxic drug preparations per year in an isolator chamber (IC). The control method is done by in-process gravimetric analysis coupled with scan identification, led by software with interactive instructions. The balances are certified once a year, yet outside the IC. Indeed, turbulent airflow could impact the scales’ measurements. The accepted errors percentages are a function of the volumes weighted.PurposeAfter software development and setup, we need to validate this control method with the two components: the weighing scales and the software.Material and methodsFor the weighting scales, a qualification was made inside and outside the IC with standard weights. For the validation the tests performed were fidelity, accuracy and eccentricity. Then, a comparison to visual control was performed to evaluate the bias of the balance. Six syringes with different volumes were made and then verified by a third person. Next, they were weighed 15 times to obtain the total error. For the software, a method is being developed to analyse the specificity and the sensitivity. For the specificity, an extraction of the software was done to study the forced steps (the steps refused by the software but accepted by the pharmacist because of the correct volume read) over a period of 6 months.ResultsThe metrological tests enable to qualify the balances. The bias of the weighing scales fluctuates between 0.94% and 4.40%. Over 6 months, 15 227 preparations were realised with a total of 1 89 334 steps including 49 180 weighing steps. Among those, there were 2023 forced steps (4.1%). The most forced cytotoxic molecules were identified. The two most forced stages were the weighing of the syringe with cytotoxic (41%) and of the final pouch (23%). The 50 ml syringe is responsible for 41% of this forced stage and, in 85% of the cases, it is because the volume to collect has a decimal value.ConclusionConcerning the sensitivity, a method is elaborated to determine the rate of the false negatives with a fake cytotoxic preparations plan and calculated weighing errors. Our method validation plan is complete with the validation of the two components: precision scale and software.References and/or acknowledgementsNo conflict of interest.

Trifolium L. species with a rich isoflavone content have been used as expectorant, analgesic, antiseptic, tonic, and wound-healer in folk medicine. The aim of the study is to evaluate pharmacological properties of the extracts and isolated compounds of T. tricocephalum. Phytochemical investigation of the aerial parts of T. trichocephalum led to the isolation of daidzein, genistein, quercetin, and daidzein 4'-O-β-glucoside for the first time from this species. Isolated compounds along with the methanol extract, water, ethyl acetate and chloroform subextracts were tested for their radical scavenging and cytotoxic activity which was evaluated by MTT assay. According to the results of activity tests, extracts showed a concentration-dependent radical scavenging activity as well as cytotoxic effect on HepG2 cells at 400 μg/mL, whereas the compounds did not exert any obvious cytotoxic effect at tested concentrations.

Marine microbes comprises approximately a half of the total global biodiversity. Sea offers an enormous resource for novel bioactive compounds and it has been classified as the largest remaining reservoir of natural molecules to be evaluated for drug activity. Marine bacteria have been received attention of researchers due to their innate potential to produce diverse compounds that attracts biological properties such as anticancer, anti-inflammatory, antimicrobial and antioxidant properties. In this study, we reported isolation of 10 strains from sea water and tested them with standard biochemical test and their potential to utilize carbon sugar, to confirm the Actinomycetes character. Among the isolates 10 and 1 was recorded to be gram positive. Carbon utilization test revealed that all the isolates utilized dextrose and none of the isolates utilized ducitol. Strain MS19 a non-motile, catalase positive, oxidase positive, and round shape strain was isolated from sea water of the Bay of Bengal.

Objective As the development and progression of colorectal cancer (CRC) are known to be affected by the immune system, cell subsets such as T cells, natural killer (NK) cells, and natural killer T (NKT) cells are considered interesting targets for immunotherapy and clinical biomarker research. Until now, the role of systemic immune profiles in tumor progression remains unclear. In this study, we aimed to characterize the immunophenotype of circulating T cells, NK cells, and NKT-like cells in patients with CRC, and to subsequently correlate these immunophenotypes to clinical follow-up data. Methods Using multiparameter flow cytometry, the subset distribution and immunophenotype of T cells (CD3 + CD56 − ), CD56 dim NK cells (CD3 − CD56 dim ), CD56 bright NK cells (CD3 − CD56 bright ), and NKT-like (CD3 + CD56 + ) cells were investigated in peripheral blood mononuclear cell (PBMC) samples from 71 CRC patients and 19 healthy donors. Results CRC patients showed profound differences in immune cell subset distribution and their immunophenotype compared to healthy donors, as characterized by increased percentage of regulatory T cells, and reduced expression level of the natural cytotoxicity receptors NKp44 and NKp46 on both CD56 dim NK cells and NKT-like cells. Finally, we showed in a multivariate analysis that above-median percentage of CD16 + NKT-like cells was independently associated with shorter disease-free survival in CRC patients. Conclusion The altered phenotype of circulating immune cell subsets in CRC and its association with clinical outcome highlight the potential use of PBMC subsets as prognostic biomarkers in CRC, thereby contributing to better insight into the role of systemic immune profiles in tumor progression.

NK cells use a variety of receptors to detect abnormal cells, including tumors and their metastases. However, in the case of melanoma, it remains to be determined what specific molecular interactions are involved and whether NK cells control metastatic progression and/or the route of dissemination. Here we show that human melanoma cell lines derived from LN metastases express ligands for natural cytotoxicity receptors (NCRs) and DNAX accessory molecule-1 (DNAM-1), two emerging NK cell receptors key for cancer cell recognition, but not NK group 2 member D (NKG2D). Compared with cell lines derived from metastases taken from other anatomical sites, LN metastases were more susceptible to NK cell lysis and preferentially targeted by adoptively transferred NK cells in a xenogeneic model of cell therapy. In mice, DNAM-1 and NCR ligands were also found on spontaneous melanomas and melanoma cell lines. Interference with DNAM-1 and NCRs by antibody blockade or genetic disruption reduced killing of melanoma cells. Taken together, these results show that DNAM-1 and NCRs are critical for NK cell-mediated innate immunity to melanoma cells and provide a background to design NK cell-based immunotherapeutic strategies against melanoma and possibly other tumors.