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Sepsis outcome is determined by a balance between inflammation and immune suppression. We aimed to evaluate monocytes polarization and reprogramming during these processes. We analyzed 93 patients with procalcitonin level >0.5 ng/mL (hPCT) and suspected/confirmed sepsis, and 84 controls by analysis of CD14, CD16 and HLA-DR expression on blood monocytes using fluorescent labeled monoclonal antibodies and BD FACS CANTO II. Complete blood cell count, procalcitonin and other biochemical markers were evaluated. Intermediate monocytes CD14++CD16+ increased in hPCT patients (including both positive and negative culture) compared to controls (13.6% ± 0.8 vs 6.2% ± 0.3, p

Il compito della fisiologia animale è quello di descrivere, analizzare e comparare, qualitativamente e quantitativamente, le funzioni che caratterizzano il mondo animale, dagli organismi unicellulari che possiedono tutte le funzioni a quelli pluricellulari più complessi la cui integrità è garantita dalle funzioni di cellule specializzate e organizzate in tessuti e organi. Il libro inizia con la presentazione delle caratteristiche chimico-fisiche dei diversi ambienti terrestri, a cui fare riferimento per capire o giustificare la comparsa di un determinato adattamento che ha reso possibile la vita in condizioni apparentemente impossibili. Viene proposta una fisiologia animale integrata con le strategie adottate per l’adattamento all’ambiente di vita, che esamina attraverso strumenti moderni e sofisticati non solo le differenze fisiologiche tra gli organismi, ma anche i fini processi e i geni che sottendono alle differenze esistenti. Lo scopo di questo libro è di offrire agli studenti universitari che seguono vari corsi di Fisiologia fonti utili di informazioni sui sistemi fisiologici e spiegazioni chiare dei concetti fisiologici facendo riferimento agli animali (vertebrati e invertebrati) nel loro ambiente naturale, non escludendo principi e concetti legati ai vari processi fisiologici per poter capire e valutare l’origine dei vari processi e considerando argomenti di attualità come la percezione degli stimoli esterni, le strategie respiratorie (la fisiologia dell’immersione, i pigmenti respiratori) o le migrazioni. Per catturare e mantenere l’interesse degli studenti sono state inserite fotografie, figure e schemi originali per illustrare e soprattutto chiarire i vari aspetti discussi nel testo. Il risultato è un testo dai contenuti molto aggiornati, con approfondimenti che originano da ricerche scientifiche molto recenti.

Alamandine is a peptide of the Renin Angiotensin System (RAS), either generated from Angiotensin A via the Angiotensin Converting Enzyme 2 (ACE2), or directly from Ang-(1–7). In mammals, it elicits cardioprotection via Mas-related G-protein-coupled receptor D (MrgD), and the NOS/NO system. In teleost fish, RAS is known to modulate heart performance. However, no information is available on the presence of a cardioactive ACE2/Alamandine axis. To fill this gap, we used the cyprinid teleost Carassius auratus (goldfish) for in silico and in vitro analyses. Via the NCBI Blast P suite we found that in cyprinids ace2 is phylogenetically detectable in a subcluster of proteins including ace2-like isoforms, and is correlated with a hypoxia-dependent pathway. By real-time PCR, Western Blotting, and HPLC, ACE2 and Alamandine were identified in goldfish heart and plasma, respectively. Both increased after chronic exposure to low O2 (2.6 mg O2 L−1). By using an ex-vivo working goldfish-heart preparation, we observed that in vitro administration of exogenous Alamandine dose-dependently stimulates myocardial contractility starting from 10−11 M. The effect that involved Mas-related receptors and PKA occurred via the NOS/NO system. This was shown by exposing the perfused heart to the NOS inhibitor L-NMMA (10−5 M) that abolished the cardiac effect of Alamandine and was supported by the increased expression of the phosphorylated NOS enzyme in the extract from goldfish heart exposed to 10−10 M Alamandine. Our data are the first to show that an ACE2/Alamandine axis is present in the goldfish C. auratus and, to elicit cardiac modulation, requires the obligatory involvement of the NOS/NO system.

Background: Molecular changes associated with the initiation of the epithelial to mesenchymal transition (EMT) program involve alterations of large proteome-based networks. The role of protein products mapping to non-coding genomic regions is still unexplored. Objective: The goal of this study was the identification of an alternative protein signature in breast cancer cellular models with a distinct expression of EMT markers. Methods: We profiled MCF-7 and MDA-MB-231 cells using liquid-chromatography mass/spectrometry (LCMS/ MS) and interrogated the OpenProt database to identify novel predicted isoforms and novel predicted proteins from alternative open reading frames (AltProts). Results: Our analysis revealed an AltProt and isoform protein signature capable of classifying the two breast cancer cell lines. Among the most highly expressed alternative proteins, we observed proteins potentially associated with inflammation, metabolism and EMT. Conclusion: Here, we present an AltProts signature associated with EMT. Further studies will be needed to define their role in cancer progression.

Nucleos(t)ide analogues (NA) belong to a family of compounds widely used in anticancer/antiviral treatments. They generally exhibit a cell toxicity limited by cellular uptake levels and the resulting nucleos(t)ides metabolism modifications, interfering with the cell machinery for nucleic acids synthesis. We previously synthesized purine nucleos(t)ide analogues N7-coordinated to a platinum centre with unaltered sugar moieties of the type: [Pt(dien)(N7-dGuo)]2+ (1; dien = diethylenetriamine; dGuo = 2'-deoxy-guanosine), [Pt(dien)(N7-dGMP)] (2; dGMP = 5'-(2'-deoxy)-guanosine monophosphate), and [Pt(dien)(N7-dGTP)]2- (3; dGTP = 5'-(2'-deoxy)-guanosine triphosphate), where the indicated electric charge is calculated at physiological pH (7.4). In this work, we specifically investigated the uptake of these complexes (1-3) at the plasma membrane level. Specific experiments on HeLa cervical cancer cells indicated a relevant cellular uptake of the model platinated deoxynucleos(t)ide 1 and 3 while complex 2 appeared unable to cross the cell plasma membrane. Obtained data buttress an uptake mechanism involving Na+-dependent concentrative transporters localized at the plasma membrane level. Consistently, 1 and 3 showed higher cytotoxicity with respect to complex 2 also suggesting selective possible applications as antiviral/antitumor drugs among the used model compounds.

Hydroxytyrosol (HT), a peculiar olive and olive oil phenolic antioxidant, plays a significant role in the endothelial and cardiovascular protection associated with olive oil consumption. However, studies examining the effects of HT on the whole-genome expression of endothelial cells, which are prominent targets for vasculo-protective effects of olive oil polyphenols, have been lacking. This study aims to comprehensively evaluate the genomic effects exerted by HT, at the transcriptional level, in endothelial cells under resting or proinflammatory conditions. Human umbilical vein endothelial cells (HUVECs) were treated with 10 µmol/L HT for 1 h and then stimulated with 5 ng/mL interleukin (IL)-1β for 3 h. Total RNA was extracted, and gene expression profile assessed with microarray analysis. Functional enrichment analysis and pathway analysis were performed by Ingenuity Pathways Analysis. Microarray data were validated by qRT-PCR. Fixing a significance threshold at 1.5-fold change, HT affected the expression of 708 and 599 genes, respectively, in HUVECs under resting and IL-1β-stimulated conditions, among these, 190 were common to both conditions. Unfolded protein response (UPR) and endoplasmic reticulum stress resulted from the two top canonical pathways common between HT and HT-IL-1β affected genes. IL-17F/A signaling was found in the top canonical pathways of HT modified genes under resting unstimulated conditions, whereas cardiac hypertrophy signaling was identified among the pathways affected by HT-IL-1β. The transcriptomic analysis allowed pinpointing immunological, inflammatory, proliferative, and metabolic-related pathways as the most affected by HT in endothelial cells. It also revealed previously unsuspected genes and related gene pathways affected by HT, thus broadening our knowledge of its biological properties. The unbiased identification of novel genes regulated by HT improves our understanding of mechanisms by which olive oil prevents or attenuates inflammatory diseases and identifies new genes to be enquired as potential contributors to the inter-individual variation in response to functional food consumption.

The Concise Guide to PHARMACOLOGY 2021/22 is the fifth in this series of biennial publications. The Concise Guide provides concise overviews, mostly in tabular format, of the key properties of nearly 1900 human drug targets with an emphasis on selective pharmacology (where available), plus links to the open access knowledgebase source of drug targets and their ligands (www.guidetopharmacology.org), which provides more detailed views of target and ligand properties. Although the Concise Guide constitutes over 500 pages, the material presented is substantially reduced compared to information and links presented on the website. It provides a permanent, citable, point-in-time record that will survive database updates. The full contents of this section can be found at http://onlinelibrary.wiley.com/doi/bph.15543. Transporters are one of the six major pharmacological targets into which the Guide is divided, with the others being: G protein-coupled receptors, ion channels, nuclear hormone receptors, catalytic receptors and enzymes. These are presented with nomenclature guidance and summary information on the best available pharmacological tools, alongside key references and suggestions for further reading. The landscape format of the Concise Guide is designed to facilitate comparison of related targets from material contemporary to mid-2021, and supersedes data presented in the 2019/20, 2017/18, 2015/16 and 2013/14 Concise Guides and previous Guides to Receptors and Channels. It is produced in close conjunction with the Nomenclature and Standards Committee of the International Union of Basic and Clinical Pharmacology (NC-IUPHAR), therefore, providing official IUPHAR classification and nomenclature for human drug targets, where appropriate.

Food intake is a vital process that supplies necessary energy and essential nutrients to the body. Information regarding luminal composition in the gastrointestinal tract (GIT) collected through mechanical and nutrient sensing mechanisms are generally conveyed, in both mammals and fish, to the hypothalamic neurocircuits. In this context, ghrelin, the only known hormone with an orexigenic action, and the intestinal peptide transporters 1 and 2, involved in absorption of dietary di- and tripeptides, exert important and also integrated roles for the nutrient uptake. Together, both are potentially involved in signaling pathways that control food intake originating from different segments of the GIT. However, little is known about the role of different paralogs and their response to fasting. Therefore, after 3 weeks of acclimatization, 12 Atlantic salmon (Salmo salar) post-smolt were fasted for 4 days to explore the gastrointestinal response in comparison with fed control (n = 12). The analysis covered morphometric (weight, length, condition factor, and wet content/weight fish %), molecular (gene expression variations), and correlation analyses. Such short-term fasting is a common and recommended practice used prior to any handling in commercial culture of the species. There were no statistical differences in length and weight but a significant lower condition factor in the fasted group. Transcriptional analysis along the gastrointestinal segments revealed a tendency of downregulation for both paralogous genes slc15a1a and slc15a1b and with significant lowered levels in the pyloric ceca for slc15a1a and in the pyloric ceca and midgut for slc15a1b. No differences were found for slc15a2a and slc15a2b (except a higher expression of the fasted group in the anterior midgut), supporting different roles for slc15 paralogs. This represents the first report on the effects of fasting on slc15a2 expressed in GIT in teleosts. Transcriptional analysis of ghrelin splicing variants (ghrl-1 and ghrl-2) showed no difference between treatments. However, correlation analysis showed that the mRNA expression for all genes (restricted to segment with the highest levels) were affected by the residual luminal content. Overall, the results show minimal effects of 4 days of induced fasting in Atlantic salmon, suggesting that more time is needed to initiate a large GIT response.

To the SLC6 family belong 20 human transporters that utilize the sodium electrochemical gradient to move biogenic amines, osmolytes, amino acids and related compounds into cells. They are classified into two functional groups, the Neurotransmitter transporters (NTT) and Nutrient amino acid transporters (NAT). Here we summarize how since their first cloning in 1998, the insect (Lepidopteran) Orthologs of the SLC6 family transporters have represented very important tools for investigating functional–structural relationships, mechanism of transport, ion and pH dependence and substate interaction of the mammalian (and human) counterparts. Supplementary Information The online version contains supplementary material available at 10.1007/s11064-021-03410-1.

Fiber photometry is widely used in neuroscience labs for in vivo detection of functional fluorescence from optical indicators of neuronal activity with a simple optical fiber. The fiber is commonly placed next to the region of interest to both excite and collect the fluorescence signal. However, the path of both excitation and fluorescence photons is altered by the uneven optical properties of the brain, due to local variation of the refractive index, different cellular types, densities and shapes. Nonetheless, the effect of the local anatomy on the actual shape and extent of the volume of tissue that interfaces with the fiber has received little attention so far. To fill this gap, we measured the size and shape of fiber photometry efficiency field in the primary motor and somatosensory cortex, in the hippocampus and in the striatum of the mouse brain, highlighting how their substructures determine the detected signal and the depth at which photons can be mined. Importantly, we show that the information on the spatial expression of the fluorescent probes alone is not sufficient to account for the contribution of local subregions to the overall collected signal, and it must be combined with the optical properties of the tissue adjacent to the fiber tip.

The increasing access to antihypertensive medications has improved longevity and quality of life in hypertensive patients. Nevertheless, hypertension still remains a major risk factor for stroke and myocardial infarction, suggesting the need to implement management of pre- and hypertensive patients. In addition to antihypertensive medications, lifestyle changes, including healthier dietary patterns, such as the Dietary Approaches to Stop Hypertension (DASH) and the Mediterranean diet, have been shown to favorably affect blood pressure and are now recommended as integrative tools in hypertension management. An analysis of the effects of nutritional components of the Mediterranean diet(s) on blood pressure has therefore become mandatory. After a literature review of the impact of Mediterranean diet(s) on cardiovascular risk factors, we here analyze the effects of olive oil and its major components on blood pressure in healthy and cardiovascular disease individuals and examine underlying mechanisms of action. Both experimental and human studies agree in showing anti-hypertensive effects of olive oil. We conclude that due to its high oleic acid and antioxidant polyphenol content, the consumption of olive oil may be advised as the optimal fat choice in the management protocols for hypertension in both healthy and cardiovascular disease patients.

A label-free localized surface plasmon resonance (LSPR)-based biosensor exploiting gold nanorods (GNRs) is proposed and demonstrated. For this aim, 35±5 nm long and 20±4 thick GNRs spaced by a few nanometers thick polyelectrolytes (PE) from a gold thin film was analyzed and synthesized. The morphology of the GNRs, the plasmon properties of GNRs, swelling of PE layers and the wettability of the surfaces were characterized by transmission and scanning electron microscopy, spectroscopic reflectivity and contact angle measurements, respectively. Indeed, when immersed in a phosphate buffer saline solution, the GNRs-PE-gold system shows an optical shift of the LSPR wavelength. This shift was found to correspond to a vertical swelling of about 2 nm, demonstrating the extreme sensitivity of the biosensor. Finally, we show that LSPR measurements can be used to detect dynamic resonance changes in response to both thickness and buffer solution, while the hydrophobic behavior of the surface can be exploited for reducing the number of liquid analytes in clinical biosensing application.

Peptide transporter 1 (PepT1) mediates the uptake of dietary di-/tripeptides in vertebrates. However, in teleost fish gut, more than one PepT1-type transporter might operate, because of teleost-specific whole gen(om)e duplication event(s) that occurred during evolution. Here, we describe a novel teleost di-/tripeptide transporter, i.e., the Atlantic salmon ( Salmo salar) peptide transporter 1a [PepT1a; or solute carrier family 15 member 1a (Slc15a1a)], which is a paralog (77% similarity and 64% identity at the amino acid level) of the well-described Atlantic salmon peptide transporter 1b [PepT1b, alias PepT1; or solute carrier family 15 member 1b (Slc15a1b)]. Comparative analysis and evolutionary relationships of gene/protein sequences were conducted after ad hoc database mining. Tissue mRNA expression analysis was performed by quantitative real-time PCR, whereas transport function analysis was accomplished by heterologous expression in Xenopus laevis oocytes and two-electrode voltage-clamp measurements. Atlantic salmon pept1a is highly expressed in the proximal intestine (pyloric ceca ≈ anterior midgut > midgut >> posterior midgut), in the same gut regions as pept1b but notably ~5-fold less abundant. Like PepT1b, Atlantic salmon PepT1a is a low‐affinity/high‐capacity system. Functional analysis showed electrogenic, Na+-independent/pH-dependent transport and apparent substrate affinity ( K0.5) values for Gly-Gln of 1.593 mmol/L at pH 7.6 and 0.076 mmol/L at pH 6.5. In summary, we show that a piscine PepT1a-type transporter is functional. Defining the role of Atlantic salmon PepT1a in the gut will help to understand the evolutionary and functional relationships among peptide transporters. Its functional characterization will contribute to elucidate the relevance of peptide transporters in Atlantic salmon nutritional physiology.

Simple Summary Current research on nanocomposite materials with tailored physical–chemical properties is increasingly advancing in biomedical applications for bone regeneration. In this study, occurrence of differential responsiveness to dextran-grafted iron oxide (DM) nanoparticles and to their hybrid nano-hydroxyapatite (DM/n-HA) counterpart was investigated in human-derived, osteoblast-like cells. Sensitivity of cells in the presence of DMs or DM/n-HAs was evaluated in terms of cytoskeletal dynamics. Remarkably, it was shown that effects triggered by the DM are no more retained when DM is embedded onto DM/n-HA nanocomposites. In parallel, analyses on the expression of genes involved in (a) intracellular signaling pathways triggered by ligands or cell interactions with elements of the extracellular matrix, (b) modulation of processes such as cell cycle arrest, apoptosis, senescence, DNA repair, metabolism changes, and (c) iron homeostasis and absorption through cell membranes, indicated that the DM/n-HA-treated cells retain tracts of physiological responsiveness unlike DM-treated cells. Overall, a shielding effect by the n-HA was assumed (masking the DM’s cytotoxicity), and a modular biomimicry of the DM/n-HA nanocomposites. On these bases, the biocompatibility of n-HA associated to DM’s magnetic responsiveness offer a combination of structural/functional features of these nano-tools for bone tissue engineering, for finely acting within physiological ranges. Abstract The development of nanocomposites with tailored physical–chemical properties, such as nanoparticles containing magnetic iron oxides for manipulating cellular events at distance, implies exciting prospects in biomedical applications for bone tissue regeneration. In this context, this study aims to emphasize the occurrence of differential responsiveness in osteoblast-like cells to different nanocomposites with diverse features: dextran-grafted iron oxide (DM) nanoparticles and their hybrid nano-hydroxyapatite (DM/n-HA) counterpart. Here, responsiveness of cells in the presence of DMs or DM/n-HAs was evaluated in terms of cytoskeletal features. We observed that effects triggered by the DM are no more retained when DM is embedded onto the DM/n-HA nanocomposites. Also, analysis of mRNA level variations of the focal adhesion kinase (FAK), P53 and SLC11A2/DMT1 human genes showed that the DM/n-HA-treated cells retain tracts of physiological responsiveness compared to the DM-treated cells. Overall, a shielding effect by the n-HA component can be assumed, masking the DM’s cytotoxic potential, also hinting a modular biomimicry of the nanocomposites respect to the physiological responses of osteoblast-like cells. In this view, the biocompatibility of n-HA together with the magnetic responsiveness of DMs represent an optimized combination of structural with functional features of the DM/n-HA nano-tools for bone tissue engineering, for finely acting within physiological ranges.

In this work, zebrafish (Danio rerio) embryos were subjected to different physical stresses, consisting of a static and two low frequency magnetic fields, and two radio frequencies. Specifically, embryos were exposed to static (Bo, 0 Hz), very low frequency (VLF, 0.2 Hz), low frequency (LF, 270 kHz), very high frequency (VHF, 100 MHz) and ultra-high frequency (UHF, 900 MHz) magnetic field irradiations for up to 5 days. The field intensities were 40 mT, 40 mT, 470 μT, 240 nT and 240 nT, respectively. Untreated embryos were used as control (n = 10 for each condition).

The feasibility to obtain highly porous collagen-based scaffolds (SCs) integrated with uniformly dispersed poly (lactide-co-glycolide)(PLGA) microparticles (MPs) was evaluated. PLGA-MPs were prepared by double emulsion technique and SCs with different amounts of MPs (SC/PLGA-MPs) were produced. Results showed that SC/PLGA-MPs could be successfully manufactured, the PLGA-MPs being physically retained in the SCs, without affecting the porous structure. PLGA-MPs also increased the SC hydrophilicity, acted as a mechanical reinforcement and retarded the degradation rate. In addition, the interactions between MPs and Caco-2 cells did not interfere with the correct morphological, adhesion and proliferation pathways.

Background Peptide transporter 1 (PepT1, alias Slc15a1) mediates the uptake of dietary di/tripeptides in all vertebrates. However, in teleost fish, more than one PepT1-type transporter might function, due to specific whole genome duplication event(s) that occurred during their evolution leading to a more complex paralogue gene repertoire than in higher vertebrates (tetrapods). Results Here, we describe a novel di/tripeptide transporter in the zebrafish (Danio rerio), i.e., the zebrafish peptide transporter 1a (PepT1a; also known as Solute carrier family 15 member a1, Slc15a1a), which is a paralogue (78% similarity, 62% identity at the amino acid level) of the previously described zebrafish peptide transporter 1b (PepT1b, alias PepT1; also known as Solute carrier family 15 member 1b, Slc15a1b). Also, we report a basic analysis of the pept1a (slc15a1a) mRNA expression levels in zebrafish adult tissues/organs and embryonic/early larval developmental stages. As assessed by expression in Xenopus laevis oocytes and two-electrode voltage clamp measurements, zebrafish PepT1a, as PepT1b, is electrogenic, Na+-independent, and pH-dependent and functions as a low-affinity system, with K0.5 values for Gly-Gln at − 60 mV of 6.92 mmol/L at pH 7.6 and 0.24 mmol/L at pH 6.5 and at − 120 mV of 3.61 mmol/L at pH 7.6 and 0.45 mmol/L at pH 6.5. Zebrafish pept1a mRNA is highly expressed in the intestine and ovary of the adult fish, while its expression in early development undergoes a complex trend over time, with pept1a mRNA being detected 1 and 2 days post-fertilization (dpf), possibly due to its occurrence in the RNA maternal pool, decreasing at 3 dpf (~ 0.5-fold) and increasing above the 1–2 dpf levels at 4 to 7 dpf, with a peak (~ 7-fold) at 6 dpf. Conclusions We show that the zebrafish PepT1a-type transporter is functional and co-expressed with pept1b (slc15a1b) in the adult fish intestine. Its expression is also confirmed during the early phases of development when the yolk syncytial layer is present and yolk protein resorption processes are active. While completing the missing information on PepT1-type transporters function in the zebrafish, these results open to future investigations on the similar/differential role(s) of PepT1a/PepT1b in zebrafish and teleost fish physiology.

Amino acids and small peptides in the form of di/tripeptides are key nutrients in animal diets and essential contributors to growth. In mammals, di/tripeptides represent a highly significant fraction of dietary protein after digestion and are transported across the brush‐border membrane of the intestinal epithelium via a single transport protein, the Peptide Transporter 1 (PepT1), a member of the Solute Carrier 15 (Slc15) family. PepT1 is a low‐affinity/high‐capacity transporter that uses an inwardly‐directed proton electrochemical gradient to drive the uptake of di/tripeptides, and peptidomimetics, across the cell membrane. However, in teleost fishes two pept1 genes, namely pept1b and pept1a, have been identified. PepT1b has been studied in many teleost species, including the Atlantic (A.) salmon (Salmo salar L.), whereas PepT1a remain unexplored. The aim of our study was to characterize, the A. salmon PepT1a vs. PepT1b in terms of tissue/organ distribution and function, and possibly extrapolate their physiological role(s). Despite the extra (4th) whole genome duplication (WGD) in the Actinopterygian lineage, A. salmon has only two pept1 genes (namely pept1a and pept1b), like in other telosts that didn't undergo the extra WGD, e.g., the zebrafish (Danio rerio). At the amino acid level, the two paralogues share 77% similarity, 64% identity. Tissue distribution and relative abundance of A. salmon pept1a and pept1b revealed that both transcripts have a similar expression profile. A. salmon pept1a is highly expressed in the proximal intestine (pyloric caeca ≈ anterior midgut > midgut >> posterior midgut), but ~5‐fold less than pept1b, which is expressed in the same gut regions. No expression signals were detected in the anterior and posterior stomach or the anterior and posterior hindgut. As assessed by heterologous expression in Xenopus laevis oocytes and Two‐Electrode Voltage Clamp measurements, A. salmon PepT1a is, like PepT1b, a low‐affinity/high‐capacity system, able to sustain an electrogenic, sodium‐independent and pH‐dependent transport of di/tripeptides. Transport associated currents show that the protein exhibits a K0.5 value for the reference dipeptide Gly‐Gln of 0.5 mM at pH 7.6 and of 0.07 mM at pH 6.5. The pre‐steady state and steady‐state transport currents analysis indicate important functional differences between A. salmon PepT1a and PepT1b transporters. The difference in expression levels and a significantly dissimilar substrate specificity for certain di/tripeptides, such as lysine‐containing peptides, supports the idea of distinct roles in peptide recognition and transport for PepT1a and PepT1b. Support or Funding Information Funded by RFFV; 247978 SalmoFeedPlus and Cargill

Zebrafish (Danio rerio) embryos were subjected to different physical stresses, consisting of a static and two low frequency magnetic fields, and two radiofrequencies. Embryos were exposed to static (Bo, 0 Hz), very low frequency (VLF, 0.2 Hz), low frequency (LF, 270 kHz), very high frequency (VHF, 100 MHz), and ultra-high frequency (UHF, 900 MHz) magnetic field irradiation for up to 5 days. The field intensities were 40 mT, 40 mT, 470 µT, 240 nT and 240 nT, respectively. Untreated embryos were used as control.(n= 10 for each condition).

Chronic inflammation of the adipose tissue (AT) is a major contributor to obesity-associated cardiometabolic complications. The olive oil polyphenol hydroxytyrosol (HT) contributes to Mediterranean diet cardiometabolic benefits through mechanisms still partially unknown. We investigated HT (1 and 10 &mu, mol/L) effects on gene expression (mRNA and microRNA) related to inflammation induced by 10 ng/mL tumor necrosis factor (TNF)-&alpha, in human Simpson&ndash, Golabi&ndash, Behmel Syndrome (SGBS) adipocytes. At real-time PCR, HT significantly inhibited TNF-&alpha, induced mRNA levels, of monocyte chemoattractant protein-1, C-X-C Motif Ligand-10, interleukin (IL)-1&beta, IL-6, vascular endothelial growth factor, plasminogen activator inhibitor-1, cyclooxygenase-2, macrophage colony-stimulating factor, matrix metalloproteinase-2, Cu/Zn superoxide dismutase-1, and glutathione peroxidase, as well as surface expression of intercellular adhesion molecule-1, and reverted the TNF-&alpha, mediated inhibition of endothelial nitric oxide synthase, peroxisome proliferator-activated receptor coactivator-1&alpha, and glucose transporter-4. We found similar effects in adipocytes stimulated by macrophage-conditioned media. Accordingly, HT significantly counteracted miR-155-5p, miR-34a-5p, and let-7c-5p expression in both cells and exosomes, and prevented NF-&kappa, B activation and production of reactive oxygen species. HT can therefore modulate adipocyte gene expression profile through mechanisms involving a reduction of oxidative stress and NF-&kappa, B inhibition. By such mechanisms, HT may blunt macrophage recruitment and improve AT inflammation, preventing the deregulation of pathways involved in obesity-related diseases.

The small intestine is the central component of the gastrointestinal (GI) tract (gut) where nutrients are absorbed into the body. Its functional structure is mainly based on its extremely extended surface area, further increased by a specific carpet of villi, responsible for the translocation of nutrients from the GI lumen into the bloodstream. Also, in the small intestine, the absorption processes of the orally administered drugs are basically related to the pharmacokinetics [1]. The deficit of cell culture methods to maintain in vivo–like functions forces researchers to optimize and apply methods in which cells are seeded and cultured under controlled and dynamic fluid flow [2]. Moreover, the lack of predictive human organ models has increased the necessity of approaches for proper mimicking of organ function in vitro, studying physiological parameters that regard mechanical, chemical and physical stimuli crucial for differentiation, morphology and function of the epithelia [3]. In this work we present a Gut-On-Chip (GOC) device, equipped with ITO (Indium tin Oxide) electrodes patterned by wet etching techniques, as a multifunctional microsystem for monitoring epithelial parameters. The potential to support cells adhesion, growth and polarization of a functional monolayer is also investigated in the Caco-2 epithelial-like cell line by in-device seeding and culture. In a perspective, this first prototype has established the basis for several technology integrations to study complex cellular phenomena targeted in key physiological topics (e.g. the tight interplay of different physical effects during mechanotransduction processes) and in pharmacological open issues such as drug absorption and metabolism.

Angiotensin II (AngII), the principal effector of the Renin-Angiotensin System, is a pluripotent humoral agent whose biological actions include short-term modulations and long-term adaptations. In fish, short-term cardio-tropic effects of AngII are documented, but information on the role of AngII in long-term cardiac remodelling is not fully understood. Here, we describe a direct approach to disclose long-term morpho-functional effects of AngII on the zebrafish heart. Adult fish exposed to waterborne teleost analogue AngII for 8 weeks showed enhanced heart weight and cardio-somatic index, coupled to myocardial structural changes (i.e. augmented compacta thickness and fibrosis), and increased heart rate. These findings were paralleled by an up-regulation of type-1 and type-2 AngII receptors expression, and by changes in the expression of GATA binding protein 4, nuclear factor of kappa light polypeptide gene enhancer in B-cells 1 and superoxide dismutase 1 soluble mRNAs, as well as of cytochrome b-245 beta polypeptide protein, indicative of cardiac remodelling. Our results suggest that waterborne AngII can sustain and robustly affect the cardiac morpho-functional remodelling of adult zebrafish.

Oxidative stress is involved in the genesis and progress of many disorders of the gastrointestinal (GI) tract. In particular, the colon epithelium is one of the GI tract segments more exposed to pro-oxidant conditions. We aimed to study the intracellular antioxidant activity of hydroxytyrosol (HT), one of the most potent natural antioxidant phenolic compounds typically present in olive oil, directly on the colon epithelium, under basal physiological and pro-oxidant conditions. Our approach was based on the application of in situ confocal microscopy on rat colon explants loaded with the fluorescent probe 5-(and-6)-chloromethyl-2’,7’-dichlorodihydrofluoresceindiacetate, which is sensitive to intracellular oxidative stress. In the intact mucosa, HT exerted a dose-dependent decrease of the basal intracellular reactive oxygen species (ROS) production of superficial colonocytes. Also, it induced a direct dose-dependent antioxidant action on the colon mucosa exposed to a pro-oxidant condition such as the H2O2 challenge. The effect of 100 µM HT was comparable to that of 10 µM Trolox, which is widely used as a standard in in vitro assays for the determination of antioxidant activity. The intracellular antioxidant activity of HT on the intact mucosa was also tested against tert-butyl peroxide, another pro-oxidant. The results show that HT can directly contribute to the redox balance of colonic epithelium by reducing ROS in both basal and pro-oxidant conditions, and support the potential of HT as a functional food ingredient with applications in protecting the intestinal mucosa against oxidative stress.

Background Recent data support the hypothesis that genetic damage occurring early in life during childhood can play an important role in the development of chronic diseases in adulthood, including cancer. Objectives The objective of this paper, part of the MAPEC_LIFE project, is to describe the frequency of micronuclei and meta-nuclear alterations in exfoliated buccal cells of 6–8year-old Italian children recruited in five Italian towns (i.e., Brescia, Torino, Pisa, Perugia and Lecce) with different air pollution levels. Methods About 200 children per town were recruited from primary schools. Biological samples were collected twice from the same children, in two different seasons (winter 2014-15 and late spring 2015). Cytogenetic damage was evaluated by the buccal micronucleus cytome assay. Results Overall,n = 1046 children represent the final cohort of the MAPEC_LIFE study. On the whole, the results showed a higher mean MN frequency in winter (0.42 ± 0.54‰) than late-spring (0.22 ± 0.34‰). MN frequency observed among the five Italian towns showed a trend that follows broadly the levels of air pollution in Italy: the highest MN frequency was observed in Brescia during both seasons, the lowest in Lecce (winter) and Perugia (late-spring). Conclusions To the best of our knowledge, the number of recruited children included in the analysis (n = 1046) is the highest compared to previous studies evaluating the frequency of MN in exfoliated buccal cells so far. MN frequency was associated with winter season and living in towns at various levels of air pollution, suggesting an important role of this exposure in determining early cytogenetic effects.

Kabuki syndrome is a rare autosomal dominant condition characterized by facial features, various organs malformations, postnatal growth deficiency and intellectual disability. The discovery of frequent germline mutations in the histone methyltransferase KMT2D and the demethylase KDM6A revealed a causative role for histone modifiers in this disease. However, the role of missense mutations has remained unexplored. Here, we expanded the mutation spectrum of KMT2D and KDM6A in KS by identifying 37 new KMT2D sequence variants. Moreover, we functionally dissected 14 KMT2D missense variants, by investigating their impact on the protein enzymatic activity and the binding to members of the WRAD complex. We demonstrate impaired H3K4 methyltransferase activity in 9 of the 14 mutant alleles and show that this reduced activity is due in part to disruption of protein complex formation. These findings have relevant implications for diagnostic and counseling purposes in this disease.

The H+-coupled peptide transporter 1 (PepT1) belongs to SoLute Carrier family 15 (SLC15A1) and is responsible for the absorption of di/tripeptides in enterocytes. Beside its nutritional role, it has been hypothesized that PepT1 functions as a transceptor, i.e. a peptide sensor/transporter involved in gut hormone release from entero-endocrine cell(s)1, 2. Studying the role of PepT1 in peptide absorption in the gut is relevant for the direct relation between dietary protein availability and fish growth3. In salmonids, PepT1 gene has been duplicated and two transporters, i.e. PepT1a and PepT1b, have been found in the intestine. The partial and complete transport cycle of the two PepT1-type transporters of Salmo salar, ssPepT1a and ssPepT1b, heterologously expressed in Xenopus laevis oocytes, were studied using Two Electrode Voltage Clamp technique. The pre-steady state currents of ssPepT1b were like that of other fish orthologs4 but differed from those of ssPepT1a. ssPepT1a showed a slower decaying currents, and the charge vs voltage (Q/V) and time constant vs voltage (τ/V) curves shifted to more positive potentials behaving as the mammalian transporter4. In both transporters, reducing external pH from 7.6 to 6.5 slowed the transients decay, shifting to more positive potential the Q/V and τ/V curves (Fig. 1). To evaluate the transport activity of ssPepT1a and ssPepT1b, the transport-associated currents were recorded in presence of 1mM of lysine(Lys)-containing peptides (KcPeps) as Lys is a limiting amino acid for animal growth3. KcPeps elicited transport-associated currents of different amplitudes, for e.g. ssPepT1b generated large currents when exposed to peptides carrying Lys in the N-terminus (KG, KM), while ssPepT1a produced small currents independently of Lys position (Fig. 2). The current vs voltage (I/V), in the presence of KcPeps, showed small and similar currents at two different pH (6.5, 7.6) in ssPepT1a. Conversely, in ssPepT1b the I/V curves differed at the most negative potentials with larger currents recorded at pH 7.6. KG dose-response experiments were also conducted, and while fitting with logistic equation allowed to obtain the kinetic parameters (K0.5 and Imax) at each voltage for ssPepT1b, only an estimation was possible for ssPepT1a at -140 and -120mV. The analysis on the transient and transport currents indicated important functional differences between ssPepT1a and ssPepT1b transporters. The dissimilar substrate specificity for KcPeps supports the idea of distinct roles in peptide recognition and transport for ssPepT1a and ssPepT1b.

The Concise Guide to PHARMACOLOGY 2017/18 is the third in this series of biennial publications. This version provides concise overviews of the key properties of nearly 1800 human drug targets with an emphasis on selective pharmacology (where available), plus links to an open access knowledgebase of drug targets and their ligands (www.guidetopharmacology.org), which provides more detailed views of target and ligand properties. Although the Concise Guide represents approximately 400 pages, the material presented is substantially reduced compared to information and links presented on the website. It provides a permanent, citable, point-in-time record that will survive database updates. The full contents of this section can be found at http://onlinelibrary.wiley.com/doi/10.1111/bph.13882/full. In addition to this overview, in which are identified 'Other protein targets' which fall outside of the subsequent categorisation, there are eight areas of focus: G protein-coupled receptors, ligand-gated ion channels, voltage-gated ion channels, other ion channels, nuclear hormone receptors, catalytic receptors, enzymes and transporters. These are presented with nomenclature guidance and summary information on the best available pharmacological tools, alongside key references and suggestions for further reading. The landscape format of the Concise Guide is designed to facilitate comparison of related targets from material contemporary to mid-2017, and supersedes data presented in the 2015/16 and 2013/14 Concise Guides and previous Guides to Receptors and Channels. It is produced in close conjunction with the Nomenclature Committee of the Union of Basic and Clinical Pharmacology (NC-IUPHAR), therefore, providing official IUPHAR classification and nomenclature for human drug targets, where appropriate.

In tissue engineering field, the production of a porous resorbable matrix, termed scaffold, allows to host cells and guide them towards the synthesis of physiological tissue. Porous scaffolds provide mechanical stability and an initial framework for migrating cells and vascular infiltration. Sustained delivery of bioactive molecules at the defect site may be also particularly important for tissue regeneration. In this context, the goal of this work was the fabrication of highly porous collagen-based scaffolds incorporating uniformly dispersed poly(lactide-co-glycolide) (PLGA) microparticles as depots for the sustained and localized delivery of bioactive molecules. Collagen scaffolds loaded with different amounts of PLGA-microparticles were prepared by freeze-drying and crosslinking. The scaffolds microstructure was assessed to evaluate the spatial distribution of microparticles and the achieved pore size. The impact of the microparticles on the scaffolds stiffness was investigated through compression tests. Preliminarily, the cell-microparticles interactions were also evaluated by imaging of cell morphology in vitro, adopting a human derived epithelial cell model. The experimental findings showed that collagen scaffolds with different amounts of uniformly dispersed PLGA-microparticles were successfully produced. The microparticles did not negatively affect the scaffold porous structure, while acting as a mechanical reinforcement. Additionally, microparticles show high permissiveness to cell adhesion, and the interactions between microparticles and epithelial cell membranes did not interfere with the correct cells morphological differentiation. Such promising results suggest the potential of the developed scaffolds for tissue engineering applications.

The zebrafish (Danio rerio) is a small vertebrate ideally suited to the modeling of human diseases. Large numbers of genetic alterations have now been modeled and could be used to study organ development by means of a genetic approach. To date, limited attention has been paid to the possible use of the zebrafish toolbox in studying human mitochondrial disorders affecting the nervous system. Here, we review the pertinent scientific literature discussing the use of zebrafish in modeling gene mutations involved in mitochondria-related neurological human diseases. A critical analysis of the literature suggests that the zebrafish not only lends itself to exploration of the pathological consequences of mitochondrial energy output on the nervous system but could also serve as an attractive platform for future drugs in an as yet untreatable category of human disorders.

Physiological mechanisms of gastrointestinal absorption of organic solutes among crustaceans remain severely underinvestigated, in spite of the considerable relevance of characterizing the routes of nutrient absorption for both nutritional purposes and formulation of balanced diets in aquaculture. Several lines of evidence attribute a primary absorptive role to the digestive gland (hepatopancreas) and a secondary role to the midgut (intestine). Among absorbed organic solutes, the importance of D-glucose in crustacean metabolism is paramount. Its plasma levels are finely tuned by hormones (crustacean hyperglycemic hormone, insulin-like peptides and insulin-like growth factors) and the function of certain organs (i.e. brain and muscle) largely depends on a balanced D-glucose supply. In the last few decades, D-glucose absorptive processes of the gastrointestinal tract of crustaceans have been described and transport mechanisms investigated, but not fully disclosed. We briefly review our present knowledge of D-glucose transport processes in the crustacean hepatopancreas. A discussion of previous results from experiments with hepatopancreatic epithelial brush-border membrane vesicles is presented. In addition, recent advances in our understandings of hepatopancreatic D-glucose transport are shown, as obtained (1) after isolation of purified R-, F-, B- and E-cell suspensions from the whole organ by centrifugal elutriation, and (2) by protein expression in hepatopancreatic mRNA-injected Xenopus laevis oocytes. In a perspective, the applicability of these novel methods to the study of hepatopancreatic absorptive function will certainly improve our knowledge of this structurally complex organ.

H+/peptide cotransport in brush-border membrane vesicles (BBMVs) from eel (Anguilla anguilla) intestine was studied by measuring D-[3H]-phenylalanyl-L-alanine uptake and by monitoring peptide-dependent intravesicular acidification using the pH-sensitive dye Acridine Orange. D-[3H]-phenylalanyl-L-alanine influx was greatly stimulated by an inside-negative membrane potential and enhanced by an inwardly directed H+ gradient. In parallel, vesicular H+ influx was significantly increased in the presence of extravesicular D-phenylalanyl-L-alanine or a series of glycyl and L-prolyl peptides. H+/peptide cotransport displayed saturable kinetics involving a single carrier system with apparent substrate affinities of 0.9–2.6 mmol l−1 depending on the particular peptide. All substrates tested competed with this system. Pre-incubation of BBMVs with dipeptides prevented diethylpyrocarbonate inhibition of transport activity, suggesting that the substrates mask histidine residues involved in the catalytic function of the transporter. Using human PepT1-specific primers, a reverse transcription–polymerase chain reaction (RT-PCR) signal was detected in eel intestine. Our results suggest that, in eel intestine, a brush-border membrane ‘low-affinity’-type H+/peptide cotransport system is present that shares kinetic features with the mammalian intestinal PepT1-type transporters.