The biosynthesis of nicotinamide adenine dinucleotide during early stages of frog embryonic development of haploid and diploid embryos.

1. Concentration of NAD during embryonic development of haploid and diploid embryos of frog was followed. NAD content in haploid embryonic forms is twice that in diploid embryos. 2. The variation of the NMN adenylyltransferase activity in the oocytes and during the first states of embryonic development as surveyed in the nuclear soluble fraction and the nuclear insoluble fraction (chromatin). 3. The enzyme activity in the soluble fraction is low during embryonic development and shows higher values in haploid embryos. 4. In the nonfertilized mature oocytes, the NMN adenylyltransferase activity is sixfold higher in the insoluble chromatin fraction than in the soluble fraction. 5. The evolution of the NMN adenylyltransferase in the insoluble chromatin fraction also shows higher values in haploid embryos, as compared with diploid forms.     

Proteomic analysis of plasma membranes isolated from undifferentiated and differentiated HepaRG cells

ABSTRACT: Liver infection with hepatitis B virus (HBV), a DNA virus of the Hepadnaviridae family, leads to severe disease, such as fibrosis, cirrhosis and hepatocellular carcinoma. The early steps of the viral life cycle are largely obscure and the host cell plasma membrane receptors are not known. HepaRG is the only proliferating cell line supporting HBV infection in vitro, following specific differentiation, allowing for investigation of new host host-cell factors involved in viral entry, within a more robust and reproducible environment. Viral infection generally begins with receptor recognition at the host cell surface, following highly specific cell-virus interactions. Most of these interactions are expected to take place at the plasma membrane of the HepaRG cells. In the present study, we used this cell line to explore changes between the plasma membrane of undifferentiated (-) and differentiated (+) cells and to identify differentially-regulated proteins or signaling networks that might potentially be involved in HBV entry. Our initial study identified a series of proteins that are differentially expressed in the plasma membrane of (-) and (+) cells and are good candidates for potential cell-virus interactions. To our knowledge, this is the first study using functional proteomics to study plasma membrane proteins from HepaRG cells, providing a platform for future experiments that will allow us to understand the cell-virus interaction and mechanism of HBV viral infection.     

Liposomalization of lactoferrin enhanced its anti-tumoral effects on melanoma cells

A number of studies have reported the anti-tumoral activity of lactoferrin, a property mediated by a variety of mechanisms such as inhibitory effects on tumor cell growth, NK cell activation, and enhancement of apoptosis. Liposomes are known to be an efficient drug delivery system which can enhance the therapeutic potential of the encapsulated compounds. We have used positively charged liposomes composed of phosphatidylcholine (PC), dioleoylphosphatidylethanolamine (DOPE), cholesterol (Chol) and stearylamine (SA) (6:1:2:1 M ratio) as a carrier system for bovine iron-free Lf (ApoBLf), and compared the in vitro effect of free and liposome-entrapped ApoBLf on the growth and morphology of murine melanoma B16-F10 cells. Liposomal formulation of ApoBLf was found to enhance the capacity of the protein to inhibit the cell proliferation by affecting cell cycle progression. The effect appeared to be due to the capacity of liposomes to increase the uptake of the protein and its accumulation into cells and probably to protect it from degradation, as revealed by fluorescence microscopy and flow cytometry. Our results demonstrate the ability of liposomes to improve the anti-tumor activity of Lf and suggest that liposomal protein may have a potential therapeutic use in the prevention and/or treatment of cancer diseases.     

Characterisation of the potamal Danube River and the Delta: connectivity determines indicative macrophyte assemblages

The complexity of water bodies in the eu-potamal river corridor and the main delta channels of the Romanian Danube is exemplified by the macrophyte vegetation. Two hypotheses provided the background for our study: (a) is the macrophyte vegetation of large, permanently connected branches significantly separated from that of the main river channel; (b) is the macrophyte composition of the Danube main stem significantly altered when the river divides into the three large navigable Delta channels. Water bodies considered were two contiguous sections of the main river channel, two large branches remaining from the historical floodplain, and the three main Delta channels. We quantified macrophyte diversity and floristic variation. Our data set was prepared from the MIDCC-project data base, in which macrophyte occurrence, abundance and habitat parameters are stored for contiguous survey units of the whole Danube river corridor. Field survey method followed that of Kohler and the European Standard EN14184. Results confirmed our first hypothesis: permanently connected side branches still support significantly different macrophyte assemblages, making them important indicators of floodplain connectivity. The diversion of the Danube into its three large navigable delta channels significantly alters the macrophyte vegetation from the c. 300 km of main stem up-river, substantially supporting our second hypothesis. Our results largely enhance the knowledge on aquatic plant biodiversity in the eu-potamal Danube, forming a solid base for long-term studies. We also discuss the relevance of our results regarding the ecological, as well as the conservational, quality of rivers and their floodplains.     

Effects of Long-Term exposure to Heavy Metals upon Rhizosphere Bacteria from Baia Mare Area (Maramureş County,Romania)

Abstract

The aim of this study was to investigate the extent of heavy metal (HM) pollution and its effect on microorganisms from rhizosphere soil in Baia Mare area (Maramure? County, Romania). Two sites with different contamination degrees were included in the study: one with a long history of mining activities and one within a drinking water safeguard zone. Rhizosphere soil samples were characterized with respect to physico-chemical parameters and the Cd, Cu, Pb and Zn contents. Native bacteria were investigated for HM tolerance and biofilm formation under toxic exposure by the microdilution assay. The most resistant strains were identified and the minimum inhibitory concentrations for HMs were determined. Cd, Cu, Pb and Zn exceeded the intervention threshold in Bozânta tailings site, while Pb content exceeded the intervention level within the area of the drinking water treatment plant. Cd showed a very high potential ecological risk in Bozânta area. The long-term exposure to HMs contributed to the selection of HM-tolerant and weakly adherent strains. Biofouling was significantly reduced under the influence of copper ions. Arthrobacter, Rhodococcus and Acidovorax strains with exceptional resistant profiles were isolated from the tailings site, indicating the important role of native microorganisms in rhizosphere ecosystems of contaminated sites.     

Towards a roadmap in brain protection and recovery

This article briefly reviews some of the mechanisms involved in the pathogenesis of neurological diseases, i.e. damage mechanisms (DM), and their interactions and overlap with protection and reparatory processes (i.e. endogenous defence activities). A relationship between DM and endogenous defence activity (EDA) regarding therapy principles will also be described. Currently, it is difficult to find the correct therapeutic approach for brain protection and recovery, especially because we do not fully understand all of the endogenous neurobiological processes, the complete nature of the pathophysiological mechanisms and the links between these two categories. Moreover, we continue to use a simplistic and reductionist approach in this respect. Endogenous neurobiological processes, such as neurotrophicity, neuroprotection, neuroplasticity and neurogenesis, are central to protection and recovery and represent the background of EDA. The biological reality of the nervous system is far more complex. In fact, there is an endogenous holistic process of neuroprotection and neurorecovery that should be approached therapeutically in an integrated way. The current tendency to exclusively frame drug activity in terms of single mechanisms and single focus effect might distract from other paradigms with greater explanatory power and hinder the development of more effective treatment strategies. A change of concept is required in pharmacological brain protection and recovery. Prospective considerations include an integrated pharmacological approach, focusing on drugs with multimodal activity and pleiotropic neuroprotective effect which are biological drugs, rather than single mechanism drugs, which usually are chemical drugs.     

TET2 is a component of the estrogen receptor complex and controls 5mC to 5hmC conversion at estrogen receptor cis-regulatory regions

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Synthesis and bioevaluation of some new isoniazid derivatives

The aim of the study was to synthesize some new compounds with potential anti-tuberculosis activity, containing isoniazid and α,β-unsaturated thiocinnamamide-like thioamides as precursors. The obtained derivatives were evaluated regarding their biological activity (antioxidant and antibacterial), as well as their influence on the eukaryotic cell cycle. The results suggested that the newly obtained derivatives of isoniazid exhibited different biological activities, depending on their structure; thus, the most active compound in terms of anti-oxidant and anti-Mycobacterium tuberculosis effects proved to be the isonicotinic acid N′-(1-amino-1-mercapto-3-phenyl-propen-1-yl)-hydrazide. This compound also increased the expression of NAT1 and NAT2 genes, which are implicated in the metabolism of the isoniazid, demonstrating that it could be rapidly metabolized, and thus well tolerated. The largest spectrum of antibacterial activity (excluding M. tuberculosis) was noticed for the isonicotinic acid N′-[1-amino-1-mercapto-3-(p-chloro-phenyl)-propen-1-yl]-hydrazide, which was also the most cytotoxic, especially at high concentrations, although not significantly affecting the cellular cycle phases. The obtained results showed that the new derivatives could represent potential candidates for the treatment of M. tuberculosis infections, but further research is needed in order to improve their pharmacological properties, by increasing their antimicrobial activity and reducing the risk of side-effects.