Expression of lamin C2 in mammalian oocytes.

Lamin C2 (LMN C2) is a short product of the lamin a gene. It is a germ cell-specific lamin and has been extensively studied in male germ cells. In this study, we focussed on the expression and localization of LMN C2 in fully-grown germinal vesicle (GV) oocytes. We detected LMN C2 in the fully-grown germinal vesicle oocytes of various mammalian species with confirmation done by immunoblotting the wild type and Lmnc2 gene deleted testes. Expression of LMN C2 tagged with GFP showed localization of LMN C2 to the nuclear membrane of the oocyte. Moreover, the LMN C2 protein notably disappeared after nuclear envelope breakdown (NEBD) and the expression of LMN C2 was significantly reduced in the oocytes from aged females and ceased altogether during meiotic maturation. These results provide new insights regarding LMN C2 expression in the oocytes of various mammalian species

Targeted mass spectrometry for monitoring of neural differentiation.

Human multipotent neural stem cells could effectively be used for the treatment of a variety of neurological disorders. However, a defining signature of neural stem cell lines that would be expandable, non-tumorigenic, and differentiate into desirable neuronal/glial phenotype after in vivo grafting is not yet defined. Employing a mass spectrometry approach, based on selected reaction monitoring, we tested a panel of well-described culture conditions, and measured levels of protein markers routinely used to probe neural differentiation, i.e. POU5F1 (OCT4), SOX2, NES, DCX, TUBB3, MAP2, S100B, GFAP, GALC, and OLIG1. Our multiplexed assay enabled us to simultaneously identify the presence of pluripotent, multipotent, and lineage-committed neural cells, thus representing a powerful tool to optimize novel and highly specific propagation and differentiation protocols. The multiplexing capacity of this method permits the addition of other newly identified cell type-specific markers to further increase the specificity and quantitative accuracy in detecting targeted cell populations. Such an expandable assay may gain the advantage over traditional antibody-based assays, and represents a method of choice for quality control of neural stem cell lines intended for clinical use

Antioxidant bioactive compounds in tomato fruits at different ripening stages and their effects on normal and cancer cells

The changes in antioxidant content of tomato fruits at different ripening stages (mature green, breaker and mature red) were determined and hydrophilic extracts were tested on normal and cancer cells. Large differences existed in the content of bioactive compounds at different ripening stages, with breaker tomatoes containing a higher content of hydrophilic antioxidants. We demonstrated a high cytotoxic effect of α-tomatine in green, but not in breaker, tomato extracts on all cell lines analysed. Cell death was found to be an apoptotic independent mechanism, probably due to α-tomatine binding to cell membrane cholesterol, disruption of cell integrity and necrosis. These results help in understanding which harvesting stage corresponds to the highest functional power of tomato fruits and may lead to the development of tomato-based functional foods