Fluorescent Labeling of Newborn Dentate Granule Cells in GAD67-GFP Transgenic Mice: A Genetic Tool for the Study of Adult Neurogenesis

Neurogenesis in the adult hippocampus is an important form of structural plasticity in the brain. Here we report a line of BAC transgenic mice (GAD67-GFP mice) that selectively and transitorily express GFP in newborn dentate granule cells of the adult hippocampus. These GFP+ cells show a high degree of colocalization with BrdU-labeled nuclei one week after BrdU injection and express the newborn neuron marker doublecortin and PSA-NCAM. Compared to mature dentate granule cells, these newborn neurons show immature morphological features: dendritic beading, fewer dendritic branches and spines. These GFP+ newborn neurons also show immature electrophysiological properties: higher input resistance, more depolarized resting membrane potentials, small and non-typical action potentials. The bright labeling of newborn neurons with GFP makes it possible to visualize the details of dendrites, which reach the outer edge of the molecular layer, and their axon (mossy fiber) terminals, which project to the CA3 region where they form synaptic boutons. GFP expression covers the whole developmental stage of newborn neurons, beginning within the first week of cell division and disappearing as newborn neurons mature, about 4 weeks postmitotic. Thus, the GAD67-GFP transgenic mice provide a useful genetic tool for studying the development and regulation of newborn dentate granule cells

Homocysteine-induced cardiomyocyte apoptosis and plasma membrane flip-flop are independent of S-adenosylhomocysteine: a crucial role for nuclear p47(phox).

Item does not contain fulltextWe previously found that homocysteine (Hcy) induced plasma membrane flip-flop, apoptosis, and necrosis in cardiomyocytes. Inactivation of flippase by Hcy induced membrane flip-flop, while apoptosis was induced via a NOX2-dependent mechanism. It has been suggested that S-adenosylhomocysteine (SAH) is the main causative factor in hyperhomocysteinemia (HHC)-induced pathogenesis of cardiovascular disease. Therefore, we evaluated whether the observed cytotoxic effect of Hcy in cardiomyocytes is SAH dependent. Rat cardiomyoblasts (H9c2 cells) were treated under different conditions: (1) non-treated control (1.5 nM intracellular SAH with 2.8 muM extracellular L -Hcy), (2) incubation with 50 muM adenosine-2,3-dialdehyde (ADA resulting in 83.5 nM intracellular SAH, and 1.6 muM extracellular L -Hcy), (3) incubation with 2.5 mM D, L -Hcy (resulting in 68 nM intracellular SAH and 1513 muM extracellular L -Hcy) with or without 10 muM reactive oxygen species (ROS)-inhibitor apocynin, and (4) incubation with 100 nM, 10 muM, and 100 muM SAH. We then determined the effect on annexin V/propodium iodide positivity, flippase activity, caspase-3 activity, intracellular NOX2 and p47(phox) expression and localization, and nuclear ROS production. In contrast to Hcy, ADA did not induce apoptosis, necrosis, or membrane flip-flop. Remarkably, both ADA and Hcy induced a significant increase in nuclear NOX2 expression. However, in contrast to ADA, Hcy additionally induced nuclear p47(phox) expression, increased nuclear ROS production, and inactivated flippase. Incubation with SAH did not have an effect on cell viability, nor on flippase activity, nor on nuclear NOX2-, p47phox expression or nuclear ROS production. HHC-induced membrane flip-flop and apoptosis in cardiomyocytes is due to increased Hcy levels and not primarily related to increased intracellular SAH, which plays a crucial role in nuclear p47(phox) translocation and subsequent ROS production.1 december 201

Outcomes from elective colorectal cancer surgery during the SARS-CoV-2 pandemic

This study aimed to describe the change in surgical practice and the impact of SARS-CoV-2 on mortality after surgical resection of colorectal cancer during the initial phases of the SARS-CoV-2 pandemic

Enterocin AS-48 inhibits the growth of – and biofilm formation by – lactic acid bacteria responsible for the accumulation of biogenic amines in cheese

Cheese is a fermented food in which toxic concentrations of biogenic amines (BAs) may accumulate, mainly as a result of the metabolism of certain lactic acid bacteria (LAB); the formation of these compounds needs to be limited. This work reports the antimicrobial potential of bacteriocin AS-48 against the LAB largely responsible for the accumulation of the BA histamine, tyramine and putrescine in cheese. Most of the 50 analysed BA-producing LAB strains were sensitive to AS-48. The bacteriocin also prevented biofilm formation of 92% of the BA-producing strains tested. This result is of great technological importance since, in cheese production, biofilms are an important source of contamination by BA-producing LAB. This study supports the potential use of AS-48 as a biopreservative for controlling BA-producing LAB in fermented food.This work was supported by the Spanish State Research Agency (AEI) and European Regional Development Fund (FEDER) (AGL2016-78708-R, AEI/FEDER, UE), by the Plan for Science, Technology and Innovation of the Principality of Asturias 2018–2020, which is co-financed by FEDER (AYUD/2021/50916, FICYT/FEDER, UE), and by the European Union's Horizon 2020 research and innovation program under the Marie Skłodowska-Curie (Grant Agreement No. 813439).Peer reviewe

High-spin states in the odd-odd N=Z nucleus 50Mn

High-spin states in the odd-odd N=Z nucleus Mn-50 have been identified. At low spin, the T=1 isobaric analogue states of Cr-50 are established up to I-pi=4(+). At high spin, the T=0 band built on the low-lying I-pi=5(+) isomer in Mn-50 becomes energetically favored and this band is observed up to its f(7/2)-shell termination at I-pi=15(+). Spherical shell model calculations in the full pf shell are found to be in excellent agreement with the experimental results. In addition, a weak side band in Mn-50 is observed to spin (12(-)) and is interpreted as a possible octupole vibrational structure. [S0556-2813(98)50211-7]