Leukocyte HMGB1 Guides Neoangiogenesis in Regenerating Skeletal Muscle

HMGB1 is ubiquitously present in the cell nucleus and highly conserved among species. It is released by necrotic cells and actively secreted by immune cells in the extracellular environment, where it behaves as a DAMP (Damage-Associate Molecular Pattern). Up to now all studies addressing the role of HMGB1 in wound healing have been carried out either in vitro or using pharmacological inhibitors. To assess the relative contribution of leukocyte HMGB1 and of passively released HMGB1 we have set up an ad hoc animal model. In this system, haematopoietic cells selectively lack HMGB1 by transplanting Hmgb1+/+ or Hmgb1-/- foetal livers into WT recipients. The role of leukocyte HMGB1 in tissue repair has been analyzed after skeletal muscle damage. I have used histological and magnetic resonance imaging (MRI) to non-invasively evaluate muscle inflammation and tissue architecture. In the absence of Hmgbl, there is a lower number of regenerating fibres at day7 after injury and an aberrant repair with smaller fibres at day 15. In vitro no differential effects of Hmgb1-/- and Hmgb1+/+ macrophages are detected on satellite cells differentiation/fusion, suggesting that events taking place in vivo are independent of the direct macrophage/satellite cell cross talk. No obvious differences are observed in the extent of leukocyte infiltration, as assessed both by cytofluorimetric and MRI analysis, suggesting that leukocyte Hmgb1 is not required for the attraction of inflammatory cells. qPCR unveils a role for leukocyte Hmgb1 in the production of angiogenic stimuli such as Ang2 and TNFα, probably in a hypoxia-dependent manner. Accordingly, the number of CD31 cells is dramatically reduced in the absence of leukocyte Hmgb1. Collectively our data suggest that HMGB1 secreted by leukocytes guides muscle regeneration through a paracrine role on vascular bed reconstitution

The nitric oxide-donor molsidomine modulates the innate inflammatory response in a mouse model of muscular dystrophy

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Diseño e implementación de un sistema de captación para el tratamiento de agua lluvia mediante ósmosis inversa para los Laboratorios de Biotecnología y Bromatología - ESPOCH.

Se realizó el diseño e implementación de un sistema de captación de agua lluvia mediante ósmosis inversa para la utilización en los laboratorios de Bioquímica y Bromatología de la Facultad de Ciencias de la Escuela Superior Politécnica de Chimborazo y dotar de agua tipo II en los laboratorios para la ejecución de prácticas estudiantiles. La investigación se fundamentó en la revisión bibliográfica de trabajos realizados con relación al tema planteado, en la caracterización física, química y microbiológica del agua lluvia y del agua tipo II obtenida mediante el tratamiento de ósmosis inversa – desionizador, en los cálculos de ingeniería, y en la construcción del sistema. Para el diseño del sistema de captación de agua lluvia se procedió a tomar valores de precipitación, intensidad perteneciente a la zona de estudio, determinándose el caudal de diseño utilizado en el cálculo de la captación de agua lluvia. El sistema cuenta con: un área de captación en los techos de los laboratorios, el sistema de conducción mediante tubería PVC, almacenamiento en un tanque de reserva de 550 L apto para abastecer el caudal de consumo de agua tipo II requerido para las prácticas estudiantiles y distribución del agua lluvia después del tratamiento de ósmosis inversa – desionizador. Con el tratamiento dado al agua de lluvia se pudo establecer que se logró reducir los parámetros físico químicos y microbiológicos del 98 al 99 %, obteniéndose una Conductividad (1 μ Siemens/cm; Cloro (3 mg/L) y Microbiológicos (10/100 mL), parámetros establecidos en la Norma ASTM D1193- 99 para agua tipo II grado analítico requerido para la utilización en los laboratorios. Con la aplicación del Sistema de captación de agua lluvia se brindará agua de excelente calidad y cantidad al sector estudiantil de la Escuela Superior Politécnica de Chimborazo para realizar las prácticas en los laboratorios de Bromatología y Bioquímica. Se recomienda realizar un mantenimiento adecuado, periódico al sistema y a los equipos implementados.The design and implementation of a system of rain water collection by reverse osmosis was used for the use in the Biochemistry and Bromatology laboratories of the Faculty of Sciences of the Polytechnic Higher School of Chimborazo and to provide type II water in the laboratories for the Execution of student practices. The research was based on a bibliographical review of works carried out in relation to the subject matter, in the physical, chemical and microbiological characterization of rainwater and type II water obtained by the treatment of reverse osmosis - deionizer, in engineering calculations, and In the construction of the system. For the design of the rainwater harvesting system, rainfall values ​​were taken, intensity belonging to the study area, determining the design flow rate used in the calculation of rainwater harvesting. The system has: a collection area in the ceilings of the laboratories, the system of conduction through PVC pipe, storage in a reserve tank of 550 L suitable to supply the type II water consumption required for student practices and Distribution of rainwater after reverse osmosis treatment - deionizer. With the treatment given to rainwater it was possible to establish that it was possible to reduce physical and microbiological parameters from 98 to 99%, obtaining a conductivity (1 μ Siemens / cm, Chlorine (3 mg / L) and Microbiological (10/100 ML), parameters established in ASTM D1193-99 for water type II analytical grade required for use in laboratories. With the application of the rainwater collection system will provide water of excellent quality and quantity to the student sector of the Higher Polytechnic School of Chimborazo to carry out the practices in the laboratories of Bromatology and Biochemistry. It is recommended to perform an adequate, periodic maintenance to the system and to the equipment implemented

Long-term monitoring for the surveillance of the conservation status of Tursiops truncatus in an EU Natura2000 site in the Mediterranean Sea. A pilot study in the Tuscan Archipelago

A pilot study, using the dataset from the research network ‘FLT Med Net’, which regularly monitors transborder regions in the Mediterranean Sea using ferries as platform for systematic surveys, was undertaken to assess common bottlenose dolphin range and population trends within the Natura 2000 EU marine site “Tutela del Tursiops truncatus”. The site was recently designated by the Tuscany Region (Italy) within the requirement of the EU Habitats Directive. In order to evaluate the conservation status of bottlenose dolphin according to the surveillance scheme of the Directive, two six-year periods (2007-2012; 2013-2018) were compared to assess trends in distribution-occurrence (range); Sightings Per Unit of Effort and Density (population). In total, 18146 NM were surveyed along two fixed transects, recording 90 sightings of Tursiops truncatus and a total of 268 specimens. Between the two periods, slight but not statistically significant differences were assessed, with decreasing trend in range and population  of the species; no variation was detected in mean group sizes. Travelling was the most common behaviour, and juveniles were present in 20% of the sightings, concentrated during spring and summer. The consistent FLT Med Net dataset was found to be appropriate to evaluate important parameters for the assessment of trends in the conservation status of Tursiops truncatus at the Natura2000 site scale.

Assessment of seasonal influenza vaccine effectiveness in patients from a central Italy reference hospital: pitfalls and intricacies from a pilot case-control study

Objectives: Influenza vaccination protects high-risk populations from severe outcomes. We assessed the feasibility of testing influenza vaccine effectiveness against hospitalization with laboratory-confirmed influenza. Methods: All hospitalized patients with influenza-like illness within 14 days, were swabbed. Cases were positive at RT-PCR for influenza A/B. Results: AtRome “GemelliHospital” (Season 2011-2012) 104 patients were contacted and 62 recruited. Considering total sample and target group (n= 47, 76%), only 29% and 38% had been vaccinated. Eighteen patients were laboratory-confirmed for influenza. Conclusions: RecruitedILI patients and prevalence of vaccinated subjects were less than expected. Larger numbers are warranted to study vaccine effectiveness against severe influenza outcomes. &nbsp

The Effect of Extensive Reading on Vocabulary Knowledge of First Level University Students

The objective of the research was to analyze how extensive reading or reading for pleasure can improve the development of vocabulary in a sample of 48 first-level university students of the Universidad Tecnológica Indoamérica in the city of Ambato. It was an experimental, bibliographic, descriptive, field and correlational research. To determine the correlation between the two study variables, a deep and thorough investigative analysis of books, journals and research was carried out. An intervention plan was applied through the use of a handbook of 19 vocabulary activities and to measure the knowledge of the vocabulary in the students, an observation card was used in order to measure attitudes and behavior towards extensive reading. A questionnaire of twelve questions related to word knowledge and the word form with its meaning was used as well. The research contains the sociodemographic statistical analysis of the participants and an analysis of the results of the evaluation of reading knowledge and vocabulary to know the descriptive aspects. Furthermore, the descriptive analysis of the vocabulary knowledge assessment from a global point of view and the aspects of word knowledge was carried out; to finally perform the analysis of the presence and degree of correlation between extensive reading and vocabulary knowledge, using the Pearson coefficient. Thus, it allowed to determine that, the existence of the correlation between the two variables investigated. That is to say, the results indicated that the use of extensive reading influences the development of students' vocabulary by raising interest and improving vocabulary

Chemical modulation of <i>in vivo</i> macrophage function with subpopulation-specific fluorescent prodrug conjugates

Immunomodulatory agents represent one of the most promising strategies for enhancing tissue regeneration without the side effects of traditional drug-based therapies. Tissue repair depends largely on macrophages, making them ideal targets for proregenerative therapies. However, given the multiple roles of macrophages in tissue homeostasis, small molecule drugs must be only active in very specific subpopulations. In this work, we have developed the first prodrug–fluorophore conjugates able to discriminate closely related subpopulations of macrophages (i.e., proinflammatory M1 vs anti-inflammatory M2 macrophages), and employed them to deplete M1 macrophages <i>in vivo</i> without affecting other cell populations. Selective intracellular activation and drug release enabled simultaneous fluorescence cell tracking and ablation of M1 macrophages <i>in vivo</i>, with the concomitant rescue of a proregenerative phenotype. <i>Ex vivo</i> assays in human monocyte-derived macrophages validate the translational potential of this novel platform to develop chemical immunomodulatory agents as targeted therapies for immune-related diseases

Alternatively activated macrophages promotes necrosis resolution following acute liver injury

Background & Aim Following acetaminophen (APAP) overdose, acute liver injury (ALI) can occur in patients that present too late for N-acetylcysteine treatment, potentially leading to acute liver failure, systemic inflammation, and death. Macrophages influence the progression and resolution of ALI due to their innate immunological function and paracrine activity. Syngeneic primary bone marrow-derived macrophages (BMDMs) were tested as a cell-based therapy in a mouse model of APAP-induced ALI (APAP-ALI). Methods Several phenotypically distinct BMDM populations were delivered intravenously to APAP-ALI mice when hepatic necrosis was established, and then evaluated based on their effects on injury, inflammation, immunity, and regeneration. In vivo phagocytosis assays were used to interrogate the phenotype and function of alternatively activated BMDMs (AAMs) post-injection. Finally, primary human AAMs sourced from healthy volunteers were evaluated in immunocompetent APAP-ALI mice. Results BMDMs rapidly localised to the liver and spleen within 4 h of administration. Injection of AAMs specifically reduced hepatocellular necrosis, HMGB1 translocation, and infiltrating neutrophils following APAP-ALI. AAM delivery also stimulated proliferation in hepatocytes and endothelium, and reduced levels of several circulating proinflammatory cytokines within 24 h. AAMs displayed a high phagocytic activity both in vitro and in injured liver tissue post-injection. Crosstalk with the host innate immune system was demonstrated by reduced infiltrating host Ly6Chi macrophages in AAM-treated mice. Importantly, therapeutic efficacy was partially recapitulated using clinical-grade primary human AAMs in immunocompetent APAP-ALI mice, underscoring the translational potential of these findings. Conclusion We identify that AAMs have value as a cell-based therapy in an experimental model of APAP-ALI. Human AAMs warrant further evaluation as a potential cell-based therapy for APAP overdose patients with established liver injury. Lay summary After an overdose of acetaminophen (paracetamol), some patients present to hospital too late for the current antidote (N-acetylcysteine) to be effective. We tested whether macrophages, an injury-responsive leukocyte that can scavenge dead/dying cells, could serve as a cell-based therapy in an experimental model of acetaminophen overdose. Injection of alternatively activated macrophages rapidly reduced liver injury and reduced several mediators of inflammation. Macrophages show promise to serve as a potential cell-based therapy for acute liver injury

Nitric Oxide Generated by Tumor-Associated Macrophages Is Responsible for Cancer Resistance to Cisplatin and Correlated With Syntaxin 4 and Acid Sphingomyelinase Inhibition

Tumor microenvironment is fundamental for cancer progression and chemoresistance. Among stromal cells tumor-associated macrophages (TAMs) represent the largest population of infiltrating inflammatory cells in malignant tumors, promoting their growth, invasion, and immune evasion. M2-polarized TAMs are endowed with the nitric oxide (NO)-generating enzyme inducible nitric oxide synthase (iNOS). NO has divergent effects on tumors, since it can either stimulate tumor cells growth or promote their death depending on the source of it; likewise the role of iNOS in cancer differs depending on the cell type. The role of NO generated by TAMs has not been investigated. Using different tumor models in vitro and in vivo we found that NO generated by iNOS of M2-polarized TAMs is able to protect tumor cells from apoptosis induced by the chemotherapeutic agent cisplatin (CDDP). Here, we demonstrate that the protective effect of NO depends on the inhibition of acid sphingomyelinase (A-SMase), which is activated by CDDP in a pathway involving the death receptor CD95. Mechanistic insights indicate that NO actions occur via generation of cyclic GMP and activation of protein kinase G (PKG), inducing phosphorylation of syntaxin 4 (synt4), a SNARE protein responsible for A-SMase trafficking and activation. Noteworthy, phosphorylation of synt4 at serine 78 by PKG is responsible for the proteasome-dependent degradation of synt4, which limits the CDDP-induced exposure of A-SMase to the plasma membrane of tumor cells. This inhibits the cytotoxic mechanism of CDDP reducing A-SMase-triggered apoptosis. This is the first demonstration that endogenous NO system is a key mechanism through which TAMs protect tumor cells from chemotherapeutic drug-induced apoptosis. The identification of the pathway responsible for A-SMase activity downregulation in tumors leading to chemoresistance warrants further investigations as a means to identify new anti-cancer molecules capable of specifically inhibiting synt4 degradation