Loss and reappearance of gap junctions in regenerating liver

Changes in intercellular junctional morphology associated with rat liver regeneration were examined in a freeze-fracture study. After a two-thirds partial hepatectomy, both gap junctions and zonulae occludentes were drastically altered. Between 0 and 20 h after partial hepatectomy, the junctions appeared virtually unchanged. 28 h after partial hepatectomy, however, the large gap junctions usually located close to the bile canaliculi and the small gap junctions enmeshed within the strands of the zonulae occudentes completely disappeared. Although the zonulae occludentes bordering the bile canaliculi apparently remained intact, numerous strands could now be found oriented perpendicular to the canaliculi. In some instances, the membrane outside the canaliculi was extensively filled with isolated junctional strands, often forming very complex configurations. About 40 h after partial hepatectomy, very many small gap junctions reappeared in close association with the zonulae occludentes. Subsequently, gap junctions increased in size and decreased in number until about 48 h after partial hepatectomy when gap junctions were indistinguishable in size and number from those of control animals. The zonulae occludentes were again predominantly located around the canalicular margins. These studies provide further evidence for the growth of gap junctions by the accretion of particles and of small gap junctions to form large maculae

Inhibition of gap junction and adherens junction assembly by connexin and A-CAM antibodies

We examined the roles of the extracellular domains of a gap junction protein and a cell adhesion molecule in gap junction and adherens junction formation by altering cell interactions with antibody Fab fragments. Using immunoblotting and immunocytochemistry we demonstrated that Novikoff cells contained the gap junction protein, connexin43 (Cx43), and the cell adhesion molecule, A-CAM (N-cadherin). Cells were dissociated in EDTA, allowed to recover, and reaggregated for 60 min in media containing Fab fragments prepared from a number of antibodies. We observed no cell-cell dye transfer 4 min after microinjection in 90% of the cell pairs treated with Fab fragments of antibodies for the first or second extracellular domain of Cx43, the second extracellular domain of connexin32 (Cx32) or A-CAM. Cell-cell dye transfer was detected within 30 s in cell pairs treated with control Fab fragments (pre-immune serum, antibodies to the rat major histocompatibility complex or the amino or carboxyl termii of Cx43). We observed no gap junctions by freeze-fracture EM and no adherens junctions by thin section EM between cells treated with the Fab fragments that blocked cell-cell dye transfer. Gap junctions were found on approximately 50% of the cells in control samples using freeze-fracture EM. We demonstrated with reaggregated Novikoff cells that: (a) functional interactions of the extracellular domains of the connexins were necessary for the formation of gap junction channels; (b) cell interactions mediated by A-CAM were required for gap junction assembly; and (c) Fab fragments of antibodies for A-CAM or connexin extracellular domains blocked adherens junction formation

Co-inoculation with Yeast and LAB Under Winery Conditions: Modification of the Aromatic Profile of Merlot Wines

The present work reports the impact of yeast/LAB co-inoculation on the aromatic profile of Merlot winesmade in wineries. This study was carried out over two consecutive years on five Merlot wines in Bordeauxand Swiss wineries, using Saccharomyces cerevisiae and Oenococcus oeni starter cultures. Seventy aromaticcompounds were quantified and, in addition, the sensory profiles of two wines were determined in orderto compare the aromatic notes of the sequential and co-inoculated wines. The influence of the timing ofinoculation with lactic acid bacteria (LAB) on the metabolic profile of wines was observed. It confirmedprevious work carried out on a micro-scale but, for the first time, the impact of yeast/LAB co-inoculationwas significantly demonstrated from a sensory point of view under winery conditions. In particular, thefruity and lactic notes, as well as the markers associated with these descriptors, such as esters and diacetyl,were altered. Co-inoculation does not always favour fruity expression, nor does it reduce the diacetylcontent and lactic aroma intensity. All of the trends were observed either in the production and degradationof metabolites, or by the development of an aromatic mask over the short and long term

Assessment of Domestic Well-Being: From Perception to Measurement

Nowadays, there are plenty of sensing devices that enable the measurement of physiological, environmental, and behavioral parameters of people 24 hours a day, seven days a week and provide huge quantities of different data. Data and signals coming from sensing devices, installed in indoor or outdoor environments or often worn by the users, generate heterogeneous and complex structured datasets, most of the time not uniformly structured. The artificial intelligence (AI) algorithms applied to these sets of data have demonstrated capabilities to infer indices related to a subject's status and well-being [1]. Well-being is a key parameter in the World Health Organization (WHO) definition of health, considering its physical, mental, and social spheres. Quantitatively assessing a subject's well-being is of paramount importance if we want to assess the whole status of a person, which is particularly useful in the case of ageing people living alone. Assessment allows for continuous remote monitoring to improve people's quality of life (QoL) according to their perceptions, needs, and preferences. Technology undoubtedly plays a pivotal role in this regard, providing us new tools to support the objective evaluation of a subject's status, including her/his perception of the living environment. Its potential is huge, also in terms of support to the healthcare system and ageing people; however, there are several engineering challenges to consider, especially in terms of sensors integrability, connectivity, and metrological performance, in order to obtain reliable and accurate measurement systems

Dielectric and optical evaluation of high-emissivity coatings for temperature measurements in microwave applications

In this work, several commercial high-emissivity coatings have been characterized in terms of emissivity, chemical composition and dielectric properties as a function of temperature, under microwave irradiation. Accurate knowledge of their response under exposure to microwaves provides new and crucial information about their practical usability for non-contact temperature measurements in microwave environments. Due to their high metallic content, some of the studied coatings exhibited unexpected microwave-triggered reactions that hindered their use up to the maximum temperature specified by the manufacturers. Emissivity and chemical analyses before and after the heating cycles confirmed the degradation of some of the samples predicted by dielectric measurements. This work illustrates how a careful characterization of optical and dielectric properties under representative operating conditions (temperature range, microwave exposure) is vital in order to select the appropriate reference coating to obtain reliable temperature measurements in microwave applications

Analyse du cis- et trans-resveratrol dans les vins produits au portugal

Nous mettons au point une méthode d'analyse du resveratrol (trallS-3,5,4'-trihydroxystilbene) à I'aide de Ia chromatographie en phase gazeuse et détection par ionisation de tlamme (FID), apres une extraction du vin par I'éther et sylilation par I'hexaméthyldisilazane (HMDS). Le seuil de détection est de 5 IIgII.Une séparation parfaite des différents isom"res tralls et eis du resveratrol est obtenue et permet leur détermination dans tous les vins étudiés. Ainsi nous montrons que I'isomere eis est toujours présent dans le vin et peut atteindre de 30 à 70 p. cent du resveratrol total. Nous présentons ici une premiere étude sur la composition en resveratrol des vins provenant de différentes régions du Portugal. Les vins analysés présentent des teneurs en isomeres trans de l'ordre de 120 à 500 Ilglldans les vins blancs, et de 400 à 4500 IIgIIdans les vins rouges. Une étude plus particuliere du Vinho Verde (vin du Nord dUPortugal) a été effectuée.Trans-resveratrol (trans-3,5,4'-trihydroxystilbene) has been reported as contributing to the health promoting properties of wine. In this study wines made exclusively from native Portuguese varieties were assayed for the levels of trans-resveratrol. The method developed uses GC-FID and GC-MS determinations of TMS derivatives; derivatisation by hexamethyldisilazane following ether extraction of wines. Two currently used food antioxydants nordihydroguaiaretic acid (NDGA) and 2' ,4' ,5' -trihydroxybutyrophenone (THBP), were employed as internai standards, being extracted and derivatised in exactly the same way as the determined resveratrol. These internal standards permitted the verification of a good linearity and reproducibility for this determination, with a detection limit of 5 μg/l. A characteristic of this method is the enhanced discrimination between cis- and trans- isomers of resveratrol with the consequent possibility of the determination of cis-resveratrol. The levels of resveratrol from wines of different regions of Portugal was studied. There appeared to be considerable variation in the concentrations encountered in the various wines although generally red wines contained much higher levels than white wines. The use of glucosidase enzymes in winemaking led to an increase of 3 - to 4 - fold in the levels of resveratrol encountered in Vinho Verde wines. Differences were noted between the resveratrol levels of specifie varietal wines from the Vinho Verde

uncertainty analysis of cell counting by metabolic assays

Cell counting is a fundamental procedure in living cell culture-based experiments and protocols in which the cell number quantification is required. The number of cells is one of the parameters necessary to investigate several cell culture features requiring to be monitored as function of time, such as cell viability, proliferation, growth, fitness and metabolism. Aim of this paper is contributing to declare a comprehensive uncertainty budget for cell counting through metabolic assays according to the EURACHEM/CITAC Guide Quantifying Uncertainty in Analytical Measurement