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Background and Purpose: ATP is highly accumulated in secretory vesicles and secreted upon exocytosis from neurons and endocrine cells. In adrenal chromaffin granules, intraluminal ATP reaches concentrations over 100 mM. However, how these large amounts of ATP contribute to exocytosis has not been investigated. Experimental Approach: Exocytotic events in bovine and mouse adrenal chromaffin cells were measured with single cell amperometry. Cytosolic Ca2+ measurements were carried out in Fluo‐4 loaded cells. Submembrane Ca2+ was examined in PC12 cells transfected with a membrane‐tethered Ca2+ indicator Lck‐GCaMP3. ATP release was measured using the luciferin/luciferase assay. Knockdown of P2X7 receptors was induced with short interfering RNA (siRNA). Direct Ca2+ influx through this receptor was measured using a P2X7 receptor‐GCamp6 construct. Key Results: ATP induced exocytosis in chromaffin cells, whereas the ectonucleotidase apyrase reduced the release events induced by the nicotinic agonist dimethylphenylpiperazinium (DMPP), high KCl, or ionomycin. The purinergic agonist BzATP also promoted a secretory response that was dependent on extracellular Ca2+. A740003, a P2X7 receptor antagonist, abolished secretory responses of these secretagogues. Exocytosis was also diminished in chromaffin cells when P2X7 receptors were silenced using siRNAs and in cells of P2X7 receptor knockout mice. In PC12 cells, DMPP induced ATP release, triggering Ca2+ influx through P2X7 receptors. Furthermore, BzATP, DMPP, and KCl allowed the formation of submembrane Ca2+ microdomains inhibited by A740003. Conclusion and Implications: Autocrine activation of P2X7 receptors constitutes a crucial feedback system that amplifies the secretion of catecholamines in chromaffin cells by favouring submembrane Ca2+ microdomains. [ABSTRACT FROM AUTHOR]

The P2X7 receptor, a member of the P2X purinergic receptor family, is a non-selective ion channel. Over the years, it has been associated with various biological functions, from modulating to regulating inflammation. However, its emerging role in antigen presentation has captured the scientific community's attention. This function is essential for the immune system to identify and respond to external threats, such as pathogens and tumor cells, through T lymphocytes. New studies show that the P2X7 receptor is crucial for controlling how antigens are presented and how T cells are activated. These studies focus on antigen-presenting cells, like dendritic cells and macrophages. This review examines how the P2X7 receptor interferes with effective antigen presentation and activates T cells and discusses the fundamental mechanisms that can affect the immune response. Understanding these P2X7-mediated processes in great detail opens up exciting opportunities to create new immunological therapies. [ABSTRACT FROM AUTHOR]

The climate emergency has prompted rapid and intensive research into sustainable, reliable, and affordable energy alternatives. Offshore wind has developed and exceeded all expectations over the last 2 decades and is now a central pillar of the UK and other international strategies to decarbonise energy systems. As the dependence on variable renewable energy resources increases, so does the importance of the necessity to develop energy storage and nonelectric energy vectors to ensure a resilient whole-energy system, also enabling difficult-to-decarbonise applications, e.g. heavy industry, heat, and certain areas of transport. Offshore wind and marine renewables have enormous potential that can never be completely utilised by the electricity system, and so green hydrogen has become a topic of increasing interest. Although numerous offshore and marine technologies are possible, the most appropriate combinations of power generation, materials and supporting structures, electrolysers, and support infrastructure and equipment depend on a wide range of factors, including the potential to maximise the use of local resources. This paper presents a critical review of contemporary offshore engineering tools and methodologies developed over many years for upstream oil and gas (O&G), maritime, and more recently offshore wind and renewable energy applications and examines how these along with recent developments in modelling and digitalisation might provide a platform to optimise green hydrogen offshore infrastructure. The key drivers and characteristics of future offshore green hydrogen systems are considered, and a SWOT (strength, weakness, opportunity, and threat) analysis is provided to aid the discussion of the challenges and opportunities for the offshore green hydrogen production sector. [ABSTRACT FROM AUTHOR]

The COVID-19 pandemic continues to affect several countries. One of the best ways to control its spread is the timely identification of infected patients for isolation and quarantine. While an episode of infection lasts an average of 8–10 days from the onset of symptoms, there is literature describing long-lasting viral persistence events. Here, we report a case of persistence of SARS-CoV-2 for 386 days in a health worker from Santiago de Chile. Our study could be one of the longest reported viral persistence events. RNA sequencing analyses indicated that the first positive diagnosis (8 June 2020) corresponded to a SARS-CoV-2 variant belonging to Clade Nextstrain 20A. Three hundred eighty-six days later (23 September 2021), the second positive result reached the same viral variant (Clade 20A) but without presence or circulation in Chile since May 2021. Both sequencing coverages showed an identity of 99.21%, with some mutations related to the severity of the disease (ORF1b:P314L) and more infectivity (S:D614G). This work reinforces the idea of implementing an RT-qPCR or rapid antigen test once the quarantine is fulfilled to ensure viral absence, identify potential persistence, and, consequently, minimize the risk of local outbreaks of SARS-CoV-2 infection. [ABSTRACT FROM AUTHOR]

The early detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) using the real-time quantitative polymerase chain reaction (RT-qPCR) as a gold-standard molecular tool has allowed to test and trace the viral spread and the isolation of COVID-19-infected patients. The detection capacity of viral and internal genes is an essential parameter to consider and analyze during the assay. In this study, we analyze the performance of the two commercial RT-qPCR kits used in Chile, TaqMan™ 2019-nCoV Control Kit v1 (Thermo Fisher) and MaxCov19 (TAAG Genetics), for the COVID-19 diagnosis from nasopharyngeal swab samples (NPSs). Our results show a lower sensitivity of the TAAG kit compared to the Thermo Fisher kit, even in the detection of SARS-CoV-2 mutations associated with its variants. This study reinforces the relevance of evaluating the performance of RT-qPCR kits before being used massively since those with lower sensitivity can generate false negatives and produce outbreaks of local infections. [ABSTRACT FROM AUTHOR]

Problem During the 1980s, Spain had very strict laws limiting access to opioid agonist maintenance treatment (OAMT). Because of this, mortality among people who used illicit opioids and other illicit drugs was high. Spain was also the European country with the highest number of cases of acquired immunodeficiency syndrome transmitted through illicit drug injection. Approach The rapid spread of human immunodeficiency virus (HIV) infection among people using heroin led to a shift from a drug-free approach to the treatment of opioid dependence to one focused on harm reduction. A substantial change in legislation made it possible to meet public health needs and offer OAMT as part of harm reduction programmes in the public health system, including prisons. Local setting Legislative changes were made throughout the country, although at a different pace in different regions. Relevant changes Legal changes facilitated the expansion of OAMT, which has achieved a coverage of 60%. A parallel reduction in the annual incidence of HIV infection has been reported. Reductions in morbidity and mortality and improved health-related quality of life have been described in patients undergoing OAMT. Lessons learnt The treatment of opioid dependence has been more heavily influenced by moral concepts and prejudices that hinder legislation and interfere with the implementation of OAMT than by scientific evidence. To fulfil public health needs, OAMT should be integrated in harm reduction programmes offered primarily in public facilities, including prisons. Longitudinal studies are needed to detect unmet needs and evaluate programme impact and suitability. Probleme Dans les annees 1980, l'Espagne avait des lois tres strictes limitant l'acces au traitement de maintien par agoniste des opioides (TMAO). Pour cette raison, la mortalite parmi les utilisateurs d'opioides illicites et autres drogues illegales etait elevee. L'Espagne etait egalement le pays europeen presentant le plus grand nombre de cas de syndrome d'immunodeficience acquise transmis par injection de drogues illicites. Approche La propagation rapide du virus de l'immunodeficience humaine (VIH) chez les personnes consommant de l'heroine a mene au basculement d'une approche non medicalisee du traitement de la dependance aux opiaces a une position axee sur la reduction des risques. Un changement important de la legislation a permis de repondre aux besoins de sante publique et d'offrir un traitement de maintien par agoniste des opioides dans le cadre des programmes de reduction des risques dans les organismes de sante publique, y compris les prisons. Environnement local Des modifications legislatives ont ete apportees dans tout le pays, bien qu'a un rythme different selon les regions. Changements significatifs Les adaptations juridiques ont facilite le developpement du traitement de maintien par agoniste des opioides, qui a atteint une couverture de 60%. Une reduction parallele de l'incidence annuelle de l'infection par le VIH a ete observee. Des reductions de morbidite et de mortalite ainsi qu'une amelioration de la qualite de vie liee a la sante ont ete decrites chez les patients suivant un tel programme. Lecons tirees Le traitement de la dependance aux opiaces a ete plus fortement influence par des concepts moraux et les prejudices qui entravent la legislation et interferent avec la mise en place de programmes de traitement que par des preuves scientifiques. Afin de repondre aux besoins de sante publique, des traitements de maintien par agoniste des opioides devraient etre integres dans les programmes de reduction des risques, majoritairement proposes dans les lieux publics, y compris les prisons. Des etudes longitudinales sont necessaires pour detecter les besoins non satisfaits et pour evaluer l'impact et la pertinence du programme. Situacion Durante la decada de los anos 80, las estrictas leyes en Espana limitaban en acceso al tratamiento de mantenimiento con opioides agonistas, por lo que la mortalidad entre los consumidores de opiaceos y otras drogas ilegales era elevada. Espana era el pais europeo con el mayor numero de casos de sindrome de la inmunodeficiencia adquirida transmitidos por la inyeccion de drogas ilegales. Enfoque La rapida propagacion de la infeccion por el virus de la inmunodeficiencia humana (VIH) entre personas que consumian heroina ocasiono un cambio de un enfoque sin farmacos para el tratamiento de la dependencia de opiaceos a otro dirigido a la reduccion de danos. Un cambio importante en la legislacion permitio satisfacer las necesidades de salud publica y ofrecer el tratamiento de mantenimiento con opioides agonistas como parte de los programas de reduccion de danos en el sistema de salud publica, incluidas las prisiones. Marco regional Se realizaron cambios legislativos en todo el pais, aunque a un ritmo distinto en las diferentes regiones. Cambios importantes Los cambios legales facilitaron la expansion del tratamiento de mantenimiento con opioides agonistas, que alcanzo una cobertura del 60 %. Se ha constatado una reduccion paralela de la incidencia de la infeccion por el VIH, y se ha evidenciado una disminucion de la morbilidad y la mortalidad, asi como una mejora en la calidad de vida entre los pacientes que se someten al tratamiento de mantenimiento con opioides agonistas. Lecciones aprendidas El tratamiento de la dependencia de opiaceos ha estado mas influido por conceptos y prejuicios morales que dificultan la legislacion e interfieren en la puesta en marcha del tratamiento de mantenimiento con opioides agonistas que por las pruebas cientificas. A fin de cumplir con las necesidades de salud publica, se deberia integrar el tratamiento de mantenimiento con opioides agonistas en los programas de reduccion de danos ofrecidos en centros publicos, incluidas las prisiones. Es necesario realizar estudios longitudinales para detectar las necesidades insatisfechas y evaluar el impacto e idoneidad del programa., Introduction The epidemic of dependence on illicit drugs, especially heroin, began in Spain in the late 1970s. By 1980, the incidence of heroin use had peaked. (1) In addition, the [...]

Personal protective measures such use of face masks, hand washing and physical distancing have proven to be effective in controlling the spread of the Covid-19 pandemic. However, adherence to these measures may have been relaxed over time. The objective of this work is to assess the change in adherence to these measures and to find factors that explain the change For this purpose, we conducted a survey in the Metropolitan Region of Chile in which we asked the adherence to these measures in August-September 2021 and retrospectively for 2020. With the answers obtained we fit a logistic regression model in which the response variable is the relaxation of each of the self-care preventive actions. The explanatory variables used are socio-demographic characteristics such as the age, sex, income, and vaccination status of the respondents. The results obtained show that there has been a significant decrease in adherence to the three personal protection measures in the Metropolitan Region of Chile. In addition, it was observed that younger people are more likely to relax these measures. The results show the importance of generating new incentives for maintaining adherence to personal protection measures. [ABSTRACT FROM AUTHOR]

The identification and tracking of SARS-CoV-2 infected patients in the general population are essential components of the global strategy to limit the COVID-19 viral spread, specifically for maintaining traceability and suppressing the resurgence of local outbreaks. Public health programs that include continuous RT-qPCR testing for COVID-19 in the general population, viral sequencing, and genomic surveillance for highly contagious forms of the virus have allowed for the identification of SARS-CoV-2 infections and reinfections. This work identified SARS-CoV-2 reinfection in a homeless person, which occurred 58 days after the first COVID-19 diagnosis. Genomic sequencing identified a different Nextstrain classification clade (20A and 20B) and PANGO lineage, with a divergence of 4 single nucleotide variants (SNVs) in S and ORF1ab genes, suggesting reinfection by different viral variants. This study is the first from the great metropolitan area of Santiago, Chile, one of the top ten countries in the world to live during the COVID-19 pandemic. We support the importance of performing intensive genomic surveillance programs in the whole population and high-risk groups, such as homeless people, nearly 20 thousand people in Chile, and have limited access to health care services and poor viral traceability. [ABSTRACT FROM AUTHOR]

Aging is associated with a deterioration of the acute phase response to inflammatory challenges. However, the nature of these defects remains poorly defined. We analyzed the hepatic inflammatory response after intraperitoneal administration of lipopolysaccharide (LPS) given to Fisher 344 rats aged 6, 15, and 22-23 months. Induction of the acute phase proteins (APPs), haptoglobin, [alpha]-1-acid glycoprotein, and T-kininogen was reduced and/or retarded with aging. Initial induction of interleukin-6 in aged rats was normal, but the later response was increased relative to younger counterparts. An exacerbated hepatic injury was observed in aged rats receiving LPS, as evidenced by the presence of multiple microabscesses in portal tracts, confluent necrosis, higher neutrophil accumulation, and elevated serum levels of alanine aminotransferase, relative to younger animals. Our results suggest that aged rats displayed a reduced expression of APPs and increased hepatic injury in response to the inflammatory insult. Key Words: Liver--Inflammation--injury--Acute phase response--Aging.

T-kininogen (T-KG) is a reliable biomarker of aging in male Sprague-Dawley rats. Here we confirm, in a longitudinal study, a similar behavior in Fisher 344 rats of both sexes. In males, the increase in serum levels of T-KG follows an exponential curve, whereas in females the increase is best fitted by a linear curve. In both genders, dietary restriction delays the increase in T-KG. We have previously shown that T-KG inhibits T lymphocyte proliferation. Here we show that serum T-KG levels correlate negatively with the ability of splenocytes (most likely B cells) to proliferate in response to lipopolysaccharide. A similar correlation was not observed with other markers of inflammation, including [alpha]1-acid glycoprotein (AGP), haptoglobin, or interleukin-10. We conclude that the increase in serum T-KG represents a useful biomarker of aging in Fisher 344, and it correlates with decreased lymphocyte proliferation with age, although a cause-effect relationship has not been established.

Kinins are vasoactive peptides released from precursors called kininogens, and serum levels of both T- and K-kinmogens increase dramatically as rats age. Kinin release is tightly regulated, and here we show that serum kinin levels also increase with age, from 63 [+ or -] 16 nmol/L in young Fisher 344 rats to 398 [+ or -] 102 nmol/L in old animals. Both K- and T-kininogens contribute sequentially to this increase, with the increase in middle-aged animals being driven primarily by K-kininogen, whereas the further augmentation in older rats occurs by increasing T-kininogen. By measuring ERK activation, we show that aorta endothelial cells from old animals are hyporesponsive to exogenous bradykinin. However, if serum kinin levels are experimentally decreased by lipopolysaccharide treatment, then the endothelial response to bradykinin is reestablished. These results indicate that serum levels of kinins increase with age, whereas the responsiveness of target cells to kinins is reduced in these same animals.

We have previously reported on a defect in both extracellular signal-regulated protein kinase (ERK) and c-jun N-terminal kinase (JNK) mitogen-activated protein kinase (MAPK) activation in splenocytes obtained from old rats. In order to investigate whether these effects are conserved across species, we have now used mouse splenocytes to measure the effect of aging on the activation of the same two MAPK families: ERK and JNK. Our results demonstrate that, as in rats, both MAPK signal transduction pathways are affected by aging in mice, indicating the existence of a further defect located downstream of the receptor-proximal events. Whereas ERK1 and [p46.sup.JNK] activation were not significantly modified, the kinetics of both ERK2 and [p54.sup.JNK] activation and inactivation were affected in splenocytes from old animals. Specifically, by analyzing the kinetics of activation and inactivation of these enzymes, we found a nearly 50% decrease in the fold of activation of both ERK2 and [p54.sup.JNK]. These defects result in an overall diminution of enzyme activities without changes in the steady-state levels of relevant proteins. The impaired activity of these two MAPK pathways is likely to play a role in the reduced expression of interleukin-2 and diminished lymphoproliferation observed in old animals.

Drug-addicted patients (N = 435) admitted for treatment in different clinical settings were studied. Patients were classified according to their self-report of consumed drugs and to the results of urine screening tests. Of the patients, 77.8% were active consumers, 9.6% were included in a methadone maintenance program, and 12.6% were abstinent. In the active consumer patients, positive urine screening results surpassed by far the information provided in the self-reports. Most patients tested positive to several drugs, while only 8.7% tested negative to all screened drugs. These results indicate that the information provided by drug-dependent patients lacks reliability when an analytical screening method is used simultaneously. Keywords - drug-addicted patients; urine screening; self-report.

Demographic, health, and socioeconomic factors significantly inform COVID-19 outcomes. This article analyzes the association of these factors and outcomes in Chile during the first five months of the pandemic. Using the municipalities Metropolitan Region's municipalities as the unit of analysis, the study looks at the role of time dynamics, space, and place in cases and deaths over a 100-day period between March and July 2020. As a result, common and idiosyncratic elements explain the prevalence and dynamics of infections and mortality. Social determinants of health, particularly multidimensional poverty index and use of public transportation play an important role in explaining differences in outcomes. The article contributes to the understanding of the determinants of COVID-19 highlighting the need to consider time-space dynamics and social determinants as key in the analysis. Structural factors are important to identify at-risk populations and to select policy strategies to prevent and mitigate the effects of COVID-19. The results are especially relevant for similar research in unequal settings. [ABSTRACT FROM AUTHOR]

Active immunotherapy against cancer is based on immune system stimulation, triggering efficient and long-lasting antigen-specific immune responses. Immunization strategies using whole dead cells from tumor tissue, containing specific antigens inside, have become a promising approach, providing efficient lymphocyte activation through dendritic cells (DCs). In this work, we generate whole dead tumor cells from CT26, E.G7, and EL4 live tumor cells as antigen sources, which termed immunogenic cell bodies (ICBs), generated by a simple and cost-efficient starvation-protocol, in order to determine whether are capable of inducing a transversal anticancer response regardless of the tumor type, in a similar way to what we describe previously with B16 melanoma. We evaluated the anticancer effects of immunization with doses of ICBs in syngeneic murine tumor models. Our results showed that mice's immunization with ICBs-E.G7 and ICBs-CT26 generate 18% and 25% of tumor-free animals, respectively. On the other hand, all carrying tumor-animals and immunized with ICBs, including ICBs-EL4, showed a significant delay in their growth compared to not immunized animals. These effects relate to DCs maturation, cytokine production, increase in CD4+T-bet+ and CD4+ROR-γt+ population, and decrease of T regulatory lymphocytes in the spleen. Altogether, our data suggest that whole dead tumor cell-based cancer immunotherapy generated by a simple starvation protocol is a promising way to develop complementary, innovative, and affordable antitumor therapies in a broad spectrum of tumors. [ABSTRACT FROM AUTHOR]

Reovirus is known to have an anticancer effect in both the preclinical and clinical assays. Current evidence suggests that the reovirus-mediated impact on tumor growth depends on the activation of specific antitumor immune responses. A feasible explanation for the oncolytic effects and immune system activation is through the expression of the fusogenic reovirus protein. In this work, we evaluated the in vivo antitumor effects of the expression of fusogenic protein p10 of avian reovirus (ARV-p10). We used chitosan nanoparticles (CH-NPs) as a vehicle for the ARV-p10 DNA in murine B16 melanoma models both in vitro and in vivo. We confirmed that ARV-p10 delivery through a chitosan-based formulation (ARV-p10 CH-NPs) was capable of inducing cell fusion in cultured melanoma cells, showing a mild cytotoxic effect. Interestingly, intratumor injection of ARV-p10 CH-NPs delayed tumor growth, without changing lymphoid populations in the tumor tissue and spleen. The injection of chitosan nanoparticles (CH-NPs) also delayed tumor growth, suggesting the nanoparticle itself would attack tumor cells. In conclusion, we proved that in vitro ARV-p10 protein expression using CH-NPs in murine melanoma cells induces a cytotoxic effect associated with its cell fusion. Further studies are necessary for establishing a protocol for efficient in vivo DNA delivery of fusion proteins to produce an antitumoral effect. [ABSTRACT FROM AUTHOR]

People use language to express their thoughts and feelings, unveiling important aspects of their psychological traits and social interactions. Although there are several studies describing methodologies to create a collection of words in English related to depression and other conditions, in most of them the selection of words is not clinical or expert based. The objective of this study is twofold: firstly, to introduce a comprehensive collection of Spanish words commonly used by patients suffering from depression, which will be available as a free open source for research purposes (GitHub), and secondly, to study the usefulness of this collection of words in identifying social media posts that could be indicative of patients suffering from depression. The level of agreement among medical doctors to determine the best words that should be used to select tweets related to depression was low. This finding may be due to the complexity of depression and the extraordinary diversity in the way people express themselves when describing their illness. It is critical to perform a thorough analysis of the specific language used in each condition, before deciding the best words to be used for filtering the tweets in each disease. As our study shows, the words supposedly more linked to depression are very common words used in other contexts, and consequently less specific for detecting depressive users. In addition, grammatical gender forms should be considered when analysing some languages such as Spanish. [ABSTRACT FROM AUTHOR]

Oncolytic virus therapy has been tested against cancer in preclinical models and clinical assays. Current evidence shows that viruses induce cytopathic effects associated with fusogenic protein-mediated syncytium formation and immunogenic cell death of eukaryotic cells. We have previously demonstrated that tumor cell bodies generated from cells expressing the fusogenic protein of the infectious salmon anemia virus (ISAV-F) enhance crosspriming and display prophylactic antitumor activity against melanoma tumors. In this work, we evaluated the effects of the expression of ISAV-F on the B16 melanoma model, both in vitro and in vivo, using chitosan nanoparticles as transfection vehicle. We confirmed that the transfection of B16 tumor cells with chitosan nanoparticles (NP-ISAV) allows the expression of a fusogenically active ISAV-F protein and decreases cell viability because of syncytium formation in vitro. However, the in vivo transfection induces a delay in tumor growth, without inducing changes on the lymphoid populations in the tumor and the spleen. Altogether, our observations show that expression of ISAV fusion protein using chitosan nanoparticles induces cell fusion in melanoma cells and slight antitumor response. [ABSTRACT FROM AUTHOR]

Melanoma immunotherapy, specifically the autotransplant of dendritic cells charged with tumors antigens, has shown promising results in clinical trials. The positive clinical effects of this therapy have been associated to increased Th17 response and delayed-type hypersensitivity (DTH) against to tumor antigens. Some synthetic compounds, such as diphenylcyclopropenone (DPCP), are capable of triggering a DTH response in cutaneous malignancies and also to induce clinically relevant effects against melanoma. In this work, we evaluated Litre extract (LExT), a standardized extract of a Chilean stinging plant, Lithraea caustic (Litre). As Litre plant is known to induce DTH, we used a murine B16 melanoma model to compare the topical and intratumor efficacy of LExT with synthetic DTH inducers (DPCP and 2,4-dinitrochlorobenzene [DNCB]). LExt contained mainly long chain catechols and sesquiterpenes. The intratumor injection of LExT induced a significant delay in tumor growth, similarly topical treatment of an established tumor with 0.1% LExT ointment induced a growth delay and even tumor regression in 15% of treated animals. No significant changes were observed on the T-cell populations associated to LExT treatment, and neither DNCB nor DPCP were capable to induce none of the LExT-induced antitumoral effects. Interestingly, our results indicate that LExT induces an antitumor response against melanoma in a mouse model and could bring a new –and affordable- treatment for melanoma in humans. [ABSTRACT FROM AUTHOR]

Second-generation antipsychotics (SGAs) are known to increase cardiovascular risk through several physiological mechanisms, including insulin resistance, hepatic steatosis, hyperphagia, and accelerated weight gain. There are limited prophylactic interventions to prevent these side effects of SGAs, in part because the molecular mechanisms underlying SGAs toxicity are not yet completely elucidated. In this perspective article, we introduce an innovative approach to study the metabolic side effects of antipsychotics through the alterations of the mitochondrial dynamics, which leads to an imbalance in mitochondrial fusion/fission ratio and to an inefficient mitochondrial phenotype of muscle cells. We believe that this approach may offer a valuable path to explain SGAs-induced alterations in metabolic homeostasis. [ABSTRACT FROM AUTHOR]

Antigen cross-presentation is a crucial step in the assembly of an antitumor immune response leading to activation of naïve CD8 T cells. This process has been extensively used in clinical trials, in which dendritic cells generated in vitro are loaded with tumor antigens and then autotransplanted to the patients. Recently, the use of autologous transplant of dendritic cells fused with dying tumor cells has demonstrated good results in clinical studies. In this work, we generated a similar process in vivo by treating mice with dead tumor cells [cell bodies (CBs)] expressing the fusogenic protein of the infectious salmon anemia virus (ISAV). ISAV fusion protein retains its fusogenic capability when is expressed on mammalian cells in vitro and the CBs expressing it facilitates DCs maturation, antigen transfer by antigen-presenting cells, and increase cross-presentation by DCs in vitro. Additionally, we observed in the melanoma model that CBs with or without ISAV fusion protein reduce tumor growth in prophylactic treatment; however, only ISAV expressing CBs showed an increase CD4 and CD8 cells in spleen. Overall, our results suggest that CBs could be used as a complement with other type of strategies to amplify antitumor immune response. [ABSTRACT FROM AUTHOR]

Neisseria gonorrhoeae (Ngo) has developed multiple immune evasion mechanisms involving the innate and adaptive immune responses. Recent findings have reported that Ngo reduces the IL-1β secretion of infected human monocyte-derived macrophages (MDM). Here, we investigate the role of adenosine triphosphate (ATP) in production and release of IL-1β in Ngo-infected MDM. We found that the exposure of Ngo-infected MDM to ATP increases IL-1β levels about ten times compared with unexposed Ngo-infected MDM (P<0.01). However, we did not observe any changes in inflammasome transcriptional activation of speck-like protein containing a caspase recruitment domain (CARD) (ASC, P>0.05) and caspase-1 (CASP1, P>0.05). In addition, ATP was not able to modify caspase-1 activity in Ngo-infected MDM but was able to increase pyroptosis (P>0.01). Notably ATP treatment defined an increase of positive staining for IL-1β with a distinctive intracellular pattern of distribution. Collectively, these data demonstrate that ATP induces IL-1β secretion by a mechanism not related to the NLRP3/ASC/caspase-1 axis and likely is acting at the level of vesicle trafficking or pore formation. [ABSTRACT FROM AUTHOR]

Human Natural Killer (NK) cells are a specialized heterogeneous subpopulation of lymphocytes involved in antitumor defense reactions. NK cell effector functions are critically dependent on cytokines and metabolic activity. Among various cytokines modulating NK cell function, interleukin-2 (IL-2) can induce a more potent cytotoxic activity defined as lymphokine activated killer activity (LAK). Our aim was to determine if IL-2 induces changes at the mitochondrial level in NK cells to support the bioenergetic demand for performing this enhanced cytotoxic activity more efficiently. Purified human NK cells were cultured with high IL-2 concentrations to develop LAK activity, which was assessed by the ability of NK cells to lyse NK-resistant Daudi cells. Here we show that, after 72 h of culture of purified human NK cells with enough IL-2 to induce LAK activity, both the mitochondrial mass and the mitochondrial membrane potential increased in a PGC-1α-dependent manner. In addition, oligomycin, an inhibitor of ATP synthase, inhibited IL-2-induced LAK activity at 48 and 72 h of culture. Moreover, the secretion of IFN-γ from NK cells with LAK activity was also partially dependent on PGC-1α expression. These results indicate that PGC-1α plays a crucial role in regulating mitochondrial function involved in the maintenance of LAK activity in human NK cells stimulated with IL-2. [ABSTRACT FROM AUTHOR]

T helper type 17 (Th17) lymphocytes, characterized by the production of interleukin-17 and other pro-inflammatory cytokines, are present in intestinal lamina propria and have been described as important players driving intestinal inflammation. Recent evidence, supporting the notion of a functional and phenotypic instability of Th17 cells, has shown that Th17 differentiate into type 1 regulatory (Tr1) T cells during the resolution of intestinal inflammation. Moreover, it has been suggested that the expression of CD39 ectonucleotidase endows Th17 cells with immunosuppressive properties. However, the exact role of CD39 ectonucleotidase in Th17 cells has not been studied in the context of intestinal inflammation. Here we show that Th17 cells expressing CD39 ectonucleotidase can hydrolyze ATP and survive to ATP-induced cell death. Moreover, in vitro-generated Th17 cells expressing the CD39 ectonucleotidase produce IL-10 and are less pathogenic than CD39 negative Th17 cells in a model of experimental colitis in Rag-/- mice. Remarkably, we show that CD39 activity regulates the conversion of Th17 cells to IL-10-producing cells in vitro, which is abrogated in the presence of ATP and the CD39-specific inhibitor ARL67156. All these data suggest that CD39 expression by Th17 cells allows the depletion of ATP and is crucial for IL-10 production and survival during the resolution of intestinal inflammation. [ABSTRACT FROM AUTHOR]

Current data suggest that Neisseria gonorrhoeae is able to suppress the protective immune response at different levels, such as B and T lymphocytes and antigen-presenting cells. The present report is focused on gonococcus evasion mechanism on macrophages (MФ) and its impact in the subsequent immune response. In response to various signals MФ may undergo classical-M1 (M1-MФ) or alternative-M2 (M2-MФ) activation. Until now there are no reports of the gonococcus effects on human MФ polarization. We assessed the phagocytic ability of monocyte-derived MФ (MDM) upon gonococcal infection by immunofluorescence and gentamicin protection experiments. Then, we evaluated cytokine profile and M1/M2 specific-surface markers on MФ challenged with N. gonorrhoeae and their proliferative effect on T cells. Our findings lead us to suggest N. gonorrhoeae stimulates a M2-MФ phenotype in which some of the M2b and none of the M1-MФ-associated markers are induced. Interestingly, N. gonorrhoeae exposure leads to upregulation of a Programmed Death Ligand 1 (PD-L1), widely known as an immunosuppressive molecule. Moreover, functional results showed that N. gonorrhoeae-treated MФ are unable to induce proliferation of human T-cells, suggesting a more likely regulatory phenotype. Taken together, our data show that N. gonorroheae interferes with MФ polarization. This study has important implications for understanding the mechanisms of clearance versus long-term persistence of N. gonorroheae infection and might be applicable for the development of new therapeutic strategies. [ABSTRACT FROM AUTHOR]

Sepsis progresses to multiple organ dysfunction due to the uncontrolled release of inflammatory mediators, and a growing body of evidence shows that neural signals play a significant role in modulating the immune response. Thus, similar to all other physiological systems, the immune system is both connected to and regulated by the central nervous system. The efferent arc consists of the activation of the hypothalamic-pituitary-adrenal axis, sympathetic activation, the cholinergic anti-inflammatory reflex, and the local release of physiological neuromodulators. Immunosensory activity is centered on the production of pro-inflammatory cytokines, signals that are conveyed to the brain through different pathways. The activation of peripheral sensory nerves, i.e., vagal paraganglia by the vagus nerve, and carotid body (CB) chemoreceptors by the carotid/sinus nerve are broadly discussed here. Despite cytokine receptor expression in vagal afferent fibers, pro-inflammatory cytokines have no significant effect on vagus nerve activity. Thus, the CB may be the source of immunosensory inputs and incoming neural signals and, in fact, sense inflammatory mediators, playing a protective role during sepsis. Considering that CB stimulation increases sympathetic activity and adrenal glucocorticoids release, the electrical stimulation of arterial chemoreceptors may be suitable therapeutic approach for regulating systemic inflammation. [ABSTRACT FROM AUTHOR]

Natural rubber latex (NRL; Hevea brasiliensis) allergy is an IgE-mediated reaction to latex proteins. When latex glove exposure is the main sensitizing agent, Hev b 5 is one of the major allergens. Dendritic cells (DC), the main antigen presenting cells, modulated with pharmacological agents can restore tolerance in several experimental models, including allergy. In the current study, we aimed to generate DC with tolerogenic properties from NRL-allergic patients and evaluate their ability to modulate allergen-specific T and B cell responses. Here we show that dexamethasone-treated DC (dxDC) differentiated into a subset of DC, characterized by low expression of MHC class II, CD40, CD80, CD86 and CD83 molecules. Compared with LPS-matured DC, dxDC secreted lower IL-12 and higher IL-10 after CD40L activation, and induced lower alloantigenic T cell proliferation. We also show that dxDC pulsed with the dominant Hev b 5 T-cell epitope peptide, Hev b 546–65, inhibited both proliferation of Hev b 5-specific T-cell lines and the production of Hev b 5-specific IgE. Additionally, dxDC induced a subpopulation of IL-10-producing regulatory T cells that suppressed proliferation of Hev b 5-primed T cells. In conclusion, dxDC generated from NRL-allergic patients can modulate allergen-specific T-cell responses and IgE production, supporting their potential use in allergen-specific immunotherapy. [ABSTRACT FROM AUTHOR]