Insulin receptor signaling and glucagon-like peptide 1 effects on pancreatic beta cells

Glucagon-like peptide-1 (GLP-1) is a potent gluco-incretin hormone, which plays a central role on pancreatic beta cell proliferation, survival and insulin secreting activity and whose analogs are used for treating hyperglycemia in type 2 diabetes mellitus. Notably, abnormal insulin signaling affects all the above-mentioned aspects on pancreatic beta cells. The aim of our study was to investigate whether the protective effects of GLP1-1 on beta cells are affected by altered insulin receptor signaling. To this end, several effects of GLP-1 were studied in INS-1E rat beta cells transfected either with an inhibitor of insulin receptor function (i.e., the Ectonucleotide Pyrophosphatase Phosphodiesterase 1, ENPP1), or with insulin receptor small interfering RNA, as well as in control cells. Crucial experiments were carried out also in a second cell line, namely the βTC-1 mouse beta cells. Our data indicate that in insulin secreting beta cells in which either ENPP1 was up-regulated or insulin receptor was down-regulated, GLP-1 positive effects on several pancreatic beta cell activities, including glucose-induced insulin secretion, cell proliferation and cell survival, were strongly reduced. Further studies are needed to understand whether such a scenario occurs also in humans and, if so, if it plays a role of clinical relevance in diabetic patients with poor responsiveness to GLP-1 related treatments

Determination of the Heavy Metal Bioaccumulation Patterns in Muscles of Two Species of Mullets from the Southern Caspian Sea

Although fish is a food that supplies nutrients of a high biological value, they can also be a source of some harmful substances, such as heavy metals. In the same context, some human activities in the Caspian Sea have contaminated this ecosystem during the past few years. For those reasons, our objective consisted of determining the concentrations of heavy metals and evaluating their bioaccumulation patterns in the different types of musculature in two species of mullets of commercial interest, Chelon auratus and Chelon saliens, from the southern coast of this sea. For this purpose, 20 C. auratus and 29 C. saliens were caught off this coastline and the metal concentrations in 3 different muscle locations were analyzed: the ventral, dorsal and caudal muscles of each fish. The caudal muscle had higher concentrations of Cd, Cu, Pb, and Zn, whereas As, Hg and Ni accumulation seemed to be independent of the musculature type. Overall, the Cd, Hg, and Pb concentrations exceeded the maximum levels permitted in fish by the European Union. In addition, the relationships between pairs of metals were positive and elevated in all the cases, which could be a sign of heavy metal pollution in the region sampled. Therefore, it will be necessary to continue monitoring and evaluating the degree of pollution in the Caspian Sea

Blood flow cytometry in Mugil cephalus and Carassius auratus: a comparative study

Blood and its parameters can give specific indications on the welfare of fishes.Many endogenous and exogenous factors exert influences on the characteristics of blood. The correct interpretation of fish hematology for a given species depends on the availability of reference values. The purpose of the present study was therefore to build a database with data on haematological profiles of Mugil cephalus (Linnaeus 1758) and goldfish Carassius auratus (Linnaeus 1758), in particular of some blood cells, namely red blood cells (RBC), white blood cells (WBC) and thrombocytes (TC). All the blood parameters studied showed significant differences in the two fish species considered, using flow cytometry and optical microscopy coupled with an automated system. In particular, RBC showed an increase in mullets in respect to goldfish, while WBC and TC decreased. The differences found may be due to the environmental conditions and the different eating habits of the two species. The results of this research will allow to better understand how the different dietary habits and environmental conditions can influence the haematological parameters of fishes. Flow cytometry represents a modern diagnostic technique in human. Moreover, the technique used by combining flow cytometry with automated haematological counting, has proved very effective in the early evaluation of haematological parameters of various fish species

Photocatalytic oxidation of aromatic alcohols to aldehydes in aqueous suspension of home-prepared titanium dioxide: 1. Selectivity enhancement by aliphatic alcohols

Oxygenated aqueous suspensions of home-prepared (HP) and commercial TiO2 catalysts were used in a batch photoreactor for carrying out the oxidation of benzyl alcohol (BA) and 4-methoxybenzyl alcohol (MBA) under different operative conditions. HP catalysts were synthesized from TiCl4 and underwent a hydrolysis treatment of different times under mild conditions. The textural characterization of catalysts was carried out with XRD, SEM observations, BET surface area and porosity measurements. For both alcohols the main oxidation products were the corresponding aromatic aldehydes and CO2. The HP catalysts exhibited selectivity values towards the aldehyde production up to 28% (BA conversion: 50%) and 41% (MBA conversion: 65%), about four times higher than those of commercial TiO2. The addition of an aliphatic alcohol (methanol, ethanol, 2-propanol or tert-butanol) in small amounts with respect to water decreased the overall oxidation rate of aromatic alcohols but enhanced the selectivity for aldehyde formation up to 1.5 times. The reactivity results suggest that: (i) the aromatic alcohol molecules interact with the TiO2 surface in different ways that eventually determine two parallel reaction pathways (partial oxidation or mineralization); (ii) the aliphatic alcohols preferentially compete with aromatic alcohols for the mineralizing pathway.Peer reviewe

Effects of pesticides on Chelon labrosus (Risso, 1827) evaluated by enzymatic activities along the north eastern Sicilian coastlines (Italy)

Pesticides are frequently applied to agricultural activities to improve harvest, in terms of yield and product quality. Useful tools for ecotoxicological studies of marine ecosystems are based on biomarker application on bioindicator key fish species. The main aim of the present study was to detect the potential presence of pesticides in a polluted coastal marine environment, namely Milazzo Gulf, situated in the north eastern coast of Sicily (Italy), by measuring the enzymatic activities of the ecotoxicological biomarkers acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) in brain and blood samples of Chelon labrosus. Also, Marinello Reserve was selected as a reference site. The data showed a significant inhibition in AChE (81%) and BChE (71%) activities in fish from Milazzo Gulf in respect to those from the reference site. The esterase inhibition is primarily due to the presence of organophosphorus insecticides and carbamates that resulted, in Milazzo Gulf, higher in concentration in respect to the reference quality standard decree (D.M. 260, 2010). The results obtained in this study confirm the suspected presence of insecticides in waters and fish from Milazzo Gulf, which may lead to a considerable hazard to humans. This study confirms the relevant advantages of the biomarker approach on fish species in the ecotoxicological evaluation of marine environments

CHK1 inhibitor sensitizes resistant colorectal cancer stem cells to nortopsentin

Limited therapeutic options are available for advanced colorectal cancer (CRC). Herein, we report that exposure to a neo-synthetic bis(indolyl)thiazole alkaloid analog, nortopsentin 234 (NORA234), leads to an initial reduction of proliferative and clonogenic potential of CRC sphere cells (CR-CSphCs), followed by an adaptive response selecting the CR-CSphC-resistant compartment. Cells spared by the treatment with NORA234 express high levels of CD44v6, associated with a constitutive activation of Wnt pathway. In CR-CSphC-based organoids, NORA234 causes a genotoxic stress paralleled by G2-M cell cycle arrest and activation of CHK1, driving the DNA damage repair of CR-CSphCs, regardless of the mutational background, microsatellite stability, and consensus molecular subtype. Synergistic combination of NORA234 and CHK1 (rabusertib) targeting is synthetic lethal inducing death of both CD44v6-negative and CD44v6-positive CRC stem cell fractions, aside from Wnt pathway activity. These data could provide a rational basis to develop an effective strategy for the treatment of patients with CRC

ATP-Dependent Infra-Slow (<0.1 Hz) Oscillations in Thalamic Networks

An increasing number of EEG and resting state fMRI studies in both humans and animals indicate that spontaneous low frequency fluctuations in cerebral activity at <0.1 Hz (infra-slow oscillations, ISOs) represent a fundamental component of brain functioning, being known to correlate with faster neuronal ensemble oscillations, regulate behavioural performance and influence seizure susceptibility. Although these oscillations have been commonly indicated to involve the thalamus their basic cellular mechanisms remain poorly understood. Here we show that various nuclei in the dorsal thalamus in vitro can express a robust ISO at ∼0.005–0.1 Hz that is greatly facilitated by activating metabotropic glutamate receptors (mGluRs) and/or Ach receptors (AchRs). This ISO is a neuronal population phenomenon which modulates faster gap junction (GJ)-dependent network oscillations, and can underlie epileptic activity when AchRs or mGluRs are stimulated excessively. In individual thalamocortical neurons the ISO is primarily shaped by rhythmic, long-lasting hyperpolarizing potentials which reflect the activation of A1 receptors, by ATP-derived adenosine, and subsequent opening of Ba2+-sensitive K+ channels. We argue that this ISO has a likely non-neuronal origin and may contribute to shaping ISOs in the intact brain

ENPP1 Affects Insulin Action and Secretion: Evidences from In Vitro Studies

The aim of this study was to deeper investigate the mechanisms through which ENPP1, a negative modulator of insulin receptor (IR) activation, plays a role on insulin signaling, insulin secretion and eventually glucose metabolism. ENPP1 cDNA (carrying either K121 or Q121 variant) was transfected in HepG2 liver-, L6 skeletal muscle- and INS1E beta-cells. Insulin-induced IR-autophosphorylation (HepG2, L6, INS1E), Akt-Ser473, ERK1/2-Thr202/Tyr204 and GSK3-beta Ser9 phosphorylation (HepG2, L6), PEPCK mRNA levels (HepG2) and 2-deoxy-D-glucose uptake (L6) was studied. GLUT 4 mRNA (L6), insulin secretion and caspase-3 activation (INS1E) were also investigated. Insulin-induced IR-autophosphorylation was decreased in HepG2-K, L6-K, INS1E-K (20%, 52% and 11% reduction vs. untransfected cells) and twice as much in HepG2-Q, L6-Q, INS1E-Q (44%, 92% and 30%). Similar data were obtained with Akt-Ser473, ERK1/2-Thr202/Tyr204 and GSK3-beta Ser9 in HepG2 and L6. Insulin-induced reduction of PEPCK mRNA was progressively lower in untransfected, HepG2-K and HepG2-Q cells (65%, 54%, 23%). Insulin-induced glucose uptake in untransfected L6 (60% increase over basal), was totally abolished in L6-K and L6-Q cells. GLUT 4 mRNA was slightly reduced in L6-K and twice as much in L6-Q (13% and 25% reduction vs. untransfected cells). Glucose-induced insulin secretion was 60% reduced in INS1E-K and almost abolished in INS1E-Q. Serum deficiency activated caspase-3 by two, three and four folds in untransfected INS1E, INS1E-K and INS1E-Q. Glyburide-induced insulin secretion was reduced by 50% in isolated human islets from homozygous QQ donors as compared to those from KK and KQ individuals. Our data clearly indicate that ENPP1, especially when the Q121 variant is operating, affects insulin signaling and glucose metabolism in skeletal muscle- and liver-cells and both function and survival of insulin secreting beta-cells, thus representing a strong pathogenic factor predisposing to insulin resistance, defective insulin secretion and glucose metabolism abnormalities

Intra-Decadal (2012–2021) Dynamics of Spatial Ichthyoplankton Distribution in Sevastopol Bay (Black Sea) Affected by Hydrometeorological Factors

High mortality of fish in the early stages of ontogenesis requires the detection of factors affecting it and is an extremely important problem. For this reason, we tried to consider the influence of some hydrometeorological factors on the intra-decadal dynamics and spatial distribution of ichthyoplankton abundance in Sevastopol Bay. To this end, we analyzed ichthyoplankton samples collected in 2012–2021 in Sevastopol Bay, and the four districts were identified. The ichthyoplankton distribution by areas was associated with the peculiarities of the pollution distribution. The maximum numbers in eggs (34.7 ± 7.5 ind. m−2) and larvae (10.0 ± 2.4 ind. m−2) were recorded in July–August. For the first time, a positive effect of temperature and repeatability of north and northeast winds on the ichthyoplankton number during the summer spawning in all identified areas of Sevastopol Bay using principal component analysis (PCA) was reported. In the southern region, positive influence of temperature and northerly winds on the ichthyoplankton abundance was confirmed by ANOVA (p = 0.02), and Pearson’s correlation analysis revealed significant correlations between these parameters on inter-annual scale (r > 0.5). In the eastern area, the discharge of rivers is an additional factor affecting the abundance of ichthyoplankton, while in the western and central areas, these factors were the wind characteristics caused by the orography peculiarities. The pollution spread in Sevastopol Bay is also largely due to the wind regime. The importance of comprehensive monitoring studies using intra-decadal data on ichthyoplankton is emphasized

Tuning the selectivity of visible light-driven hydroxylation of benzene to phenol by using Cu, Fe and V oxides supported on N-doped TiO2

Cu, Fe and V oxides supported on N-TiO2 (Cu/N-TiO2, Fe/N-TiO2, and V/N-TiO2) were synthesized by an incipient wet impregnation method. The prepared photocatalysts were analyzed by N-2 adsorption at -196 degrees C to measure the specific surface area (S-BET) values, scanning electron microscopy (SEM), wide-angle X-ray diffraction (WAXD), X-ray photoelectron spectroscopy (XPS), and Raman, photoluminescence and ultraviolet-visible diffuse reflectance (UV-vis DRS) spectroscopy methods. The prepared photocatalysts were tested in the hydroxylation of benzene to phenol under visible light irradiation in the presence of H2O2 as the oxidant. After 360 min of irradiation, Cu/N-TiO2 achieves a phenol yield equal to 25%, significantly higher than that observed with Fe/N-TiO2 (2%) and V/N-TiO2 (2.5%). Moreover, the Cu/N-TiO2 photocatalyst exhibited a phenol yield higher than that reported in the literature dealing with TiO2 based photocatalysts for photocatalytic benzene hydroxylation. The better photoactivity of Cu/N-TiO2 in phenol production was justified by considering both electronic and surface photocatalyst features. In detail, a significant optical absorption in the visible region has been highlighted, due to the intense electronic interactions between CuO and N-TiO2. Moreover, the surface of the copper oxide component shows low affinity with phenol molecules. Therefore, once photocatalytically generated, phenol easily desorbs from the Cu/N-TiO2 surface, thus limiting parasitic overoxidation reactions. In fact, after 180 min of visible light irradiation, only 30% of phenol was degraded by Cu/N-TiO2, while 100% and 81% of it was degraded by Fe/N-TiO2 and V/N-TiO2, respectively. The comparison of phenol production kinetic constants, obtained by fitting the experimental data with the least-squares methods, showed that the highest rate of phenol formation (k = 1.41 x 10(-3) min(-1)) was obtained by using the Cu/N-TiO2 photocatalyst. Cu/N-TiO2 has been recovered from the aqueous solution after a photocatalytic run and reused four times with no reduction in benzene conversion and phenol yield, thus confirming the high stability of the catalytic system