A characterization of four B16 murine melanoma cell sublines molecular fingerprint and proliferation behavior

Background: One of the most popular and versatile model of murine melanoma is by inoculating B16 cells in the syngeneic C57BL6J mouse strain. A characterization of different B16 modified cell sub-lines will be of real practical interest. For this aim, modern analytical tools like surface enhanced Raman spectroscopy/scattering (SERS) and MTT were employed to characterize both chemical composition and proliferation behavior of the selected cells. Methods: High quality SERS signal was recorded from each of the four types of B16 cell sub-lines: B164A5, B16GMCSF, B16FLT3, B16F10, in order to observe the differences between a parent cell line (B164A5) and other derived B16 cell sub-lines. Cells were incubated with silver nanoparticles of 50–100 nm diameter and the nanoparticles uptake inside the cells cytoplasm was proved by transmission electron microscopy (TEM) investigations. In order to characterize proliferation, growth curves of the four B16 cell lines, using different cell numbers and FCS concentration were obtained employing the MTT proliferation assay. For correlations doubling time were calculated. Results: SERS bands allowed the identification inside the cells of the main bio-molecular components such as: proteins, nucleic acids, and lipids. An "on and off" SERS effect was constantly present, which may be explained in terms of the employed laser power, as well as the possible different orientations of the adsorbed species in the cells in respect to the Ag nanoparticles. MTT results showed that among the four tested cell sub-lines B16 F10 is the most proliferative and B164A5 has the lower growth capacity. Regarding B16FLT3 cells and B16GMCSF cells, they present proliferation ability in between with slight slower potency for B16GMCSF cells. Conclusion: Molecular fingerprint and proliferation behavior of four B16 melanoma cell sub-lines were elucidated by associating SERS investigations with MTT proliferation assay

Maintaining intestinal microflora balance in heat-stressed broilers using dietary creeping wood sorrel (Oxalis corniculata) powder and chromium (chromium picolinate)

This study aimed to determine the effect of dietary creeping wood sorrel powder (Oxalis corniculata) and chromium supplemented to broilers (1-42 days) exposed to heat stress, on their performance and on the intestinal and caecal microbiota.Area of study: Ilfov, RomaniaMaterial and methods: The feeding trial was conducted on 60, day-old Cobb 500 broilers, divided equally in two groups, each group with six replicates (5 chicks/ replicate). The broilers were housed in an experimental hall at 32° C constant temperature and 23h light regimen. Unlike the dietary control diet (C), the experimental diet (E) was supplemented with 1% creeping wood sorrel powder and 20 mg chromium picolinate/ kg premix. One bird from each replication was slaughtered on days 28 and 42, and samples of caecal and intestinal content were collected for bacteriological analysis.Main results: The dietary creeping wood sorrel powder and chromium supplements for heat-stressed broilers had no significant influence on their growth performance (1-42 d). Overall, E diet had a beneficial effect on the balance of the caecal microflora; however, in the intestine, E diet had a positive influence on the balance of the intestinal microflora, only for the samples collected at 28 days.Research highlights: Dietary creeping wood sorrel powder and chromium supplements can be an efficient tool for maintaining a proper balance of intestinal microflora in heat-stressed broilers.

An Update On Natural Compounds and Their Modern Formulations for the Management of Malignant Melanoma

The chapter includes a brief presentation of the types of skin cancer. The most aggressive type of skin cancer, melanoma, is discussed from the point of view of incidence, molecular, and immunohistochemical mechanism along with the most important biomarkers for identification. Recent studies containing active phytocompounds with chemopreventive activity pointing toward phytochemicals used for melanoma prevention and therapy are reviewed. Modern physicochemical formulations for the enhancement of bioavailability of some active phytochemicals with chemopreventive activity for malignant melanoma are discussed

Total phenolic content, FTIR analysis, and antiproliferative evaluation of lupin seeds harvest from western Romania

Introduction Lupinus spp. are herbaceous perennial flowering plants included in the Fabaceae family. Among the approximately 200 species belonging to this genre, Lupinus albus L., also known as white lupin, Lupinus angustifolius L., and narrow-leafed lupin or blue lupin, represent two of the most studied species due to their intense culinary use and potential biological activity. The intention of the study was to characterize total phenolic content, perform FTIR analysis, and antiproliferative effects against A375 human melanoma cells extracts obtained from germinated and ungerminated seeds from Lupinus albus L. and Lupinus angustifolius L. Material and methods Total phenolic content was assessed using the Folin-Ciocalteu colorimetric method. FTIR spectra were carried out by a Shimadzu Prestige-21 spectrometer in the range 400–4000 cm-1, using KBr pellets and resolution of 4 cm-1. Antiproliferative capacity was screened by employing the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and scratch assay methods. Results The study showed increased values corresponding to total phenolic content for the germinated extracts. FTIR spectra confirmed the presence of genistein and main cinnamic acids derivatives (ferulic, caffeic, rosmarinic, and coumaric acids). All tested extracts showed weak antiproliferative potential against A375 human melanoma cells. Conclusions Germination increased the amount of bioactive compounds in the seeds of the two studied species of lupin. FTIR analyses provided an important fingerprint of the chemical composition

Preclinical Aspects on Magnetic Iron Oxide Nanoparticles and Their Interventions as Anticancer Agents: Enucleation, Apoptosis and Other Mechanism

The broad area of magnetic iron oxide nanoparticle (M-IONP) applications and their exclusive physico-chemical characteristics (superparamagnetic properties per se, solubility and stability in aqueous solutions, and high bioavailability in vivo) make these nanoparticles suitable candidates for biomedical uses. The most employed magnetic iron oxides in the biomedical field are magnetite and maghemite. Cancer represents a complex pathology that implies multiple mechanisms and signaling pathways, this complexity being responsible for the increased resistance to therapy and the lack of an effective curative treatment. A potential useful alternative was considered to be the use of magnetic iron nanoparticles. The M-IONPs proved to be effective as contrast agents in magnetic resonance imaging, as drug delivery carriers for different therapeutic agents, in magnetic cell separation assays, and are suitable to be engineered in terms of size, targeted delivery and substance release. Moreover, their in vivo administration was considered safe, and recent studies indicated their efficiency as anticancer agents. This chapter aims to furnish an overview regarding the physico-chemical properties of M-IONPs (mainly magnetite, maghemite and hematite), the synthesis methods and their in vitro biological impact on healthy and cancer cell lines, by describing their potential mechanism of action—enucleation, apoptosis or other mechanisms

Phytocompounds Targeting Cancer Angiogenesis Using the Chorioallantoic Membrane Assay

Cancer is the second cause of mortality worldwide. Angiogenesis is an important process involved in the growth of primary tumors and metastasis. New approaches for controlling the cancer progression and invasiveness can be addressed by limiting the angiogenesis process. An increasingly large number of natural compounds are evaluated as angiogenesis inhibitors. The chorioallantoic membrane (CAM) assay represents an in vivo attractive experimental model for cancer and angiogenesis studies as prescreening to the murine models. Since the discovery of tumor angiogenesis, the CAM has been intensively used in cancer research. The advantages of this in vivo technique are in terms of low time-consuming, costs, and a lower number of sacrificed animals. Currently, a great number of natural compounds are being investigated for their effectiveness in controlling tumor angiogenesis. Potential reducing of angiogenesis has been investigated by our group for pentacyclic triterpenes, in various formulations, and differences in their mechanism were registered. This chapter aims to give an overview on a number of phytocompounds investigated using in vitro, murine models and the chorioallantoic membrane assay as well as to emphasize the use of CAM assay in the study of natural compounds with potential effects in malignancies

Cardioprotective Effects of Cultivated Black Chokeberries (<em>Aronia</em> spp.): Traditional Uses, Phytochemistry and Therapeutic Effects

Cardiovascular diseases represent the main cause of morbidity and mortality worldwide. Obesity, sedentary life style, diet, smoking and stress are the principal inducers of hypertension, endothelium dysfunction and insulin resistance in the developed countries. The latest in vitro and in vivo studies on different type of extracts obtained from black-fruited Aronia highlight its excellent cardioprotective actions for the prevention and treatment of cardiovascular and metabolic disorders. So, this chapter aims to bring an up-to-date regarding the antioxidant, anti-inflammatory, anti-atherosclerotic, antiplatelet, blood pressure, glucose and lipid reduction properties of black-fruited Aronia, as a possible new therapeutic strategy for the primary and secondary prevention of cardiovascular pathologies