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Introduction and Objectives: International studies have described an epidemiological change in patients with the hepatitis C virus (HCV), with greater involvement of young people and risk groups. The reality in Latin America, particularly in Chile, is unknown. This study aimed to epidemiologically characterize HCV patients treated in the Chilean public health system (period 2016-2021) and compare these characteristics in two periods (2016 - 2019 vs. 2020 - 2021). Materials and Methods: Historical cohort was constructed based on national data and the Hospital del Salvador registry (Santiago, Chile) (n=410). All patients diagnosed with HCV treated in the Chilean public system (2016-2019) and those treated at Hospital del Salvador (2020-2021 period) were included. It was registered: year of diagnosis, age, sex, presence of cirrhosis, HCV genotype, co-infection with hepatitis B virus (HBV) and/or HIV, need for a liver transplant, or intratreatment dead. Both periods were compared using the Mann-Whitney U test or Fisher's exact test, as appropriate. Results: 61.2% of patients were male, with a median age of 57 years. 73.5% presented genotype 1 and 11.6% genotype 4. There was a 19.3% coinfection with HIV. Only 1.4% had therapy failure at 24 weeks and 5.2% of patients underwent liver transplantation. When comparing the periods 2016 – 2019 vs. 2020 – 2021 a reduction in the median age 59 vs 49 (p

Introduction and Objectives: Acute-on-chronic liver failure (ACLF) is a severe clinical entity with organ failures and high short-term mortality. To Date, few ACLF reports have been published in Latin America. This study aimed to characterize patients with ACLF, identifying triggers, organ failure, and survival at 30, 90, and 180 days compared to patients with decompensated cirrhosis without ACLF. Materials and Methods: Retrospective study of decompensated cirrhotic patients hospitalized (between 2017-2019) in three centers in Chile. We evaluated transplant-free survival using Kaplan-Meier curves and Cox-regression. Results: 398 patients were included, a median age of 65.3±11.7-year-old, 50.5% female, 91 (22.9%) presented ACLF (8% ACLF-1, 6.3% ACLF-2, 8.6% ACLF-3); 6.6% underwent liver transplantation. ACLF patients were younger (63.6 vs. 66.0 years; p=0.045), had higher MELD-Na scores (27 [23-32] vs. 17 [13-23]; p

Introduction and Objectives: Hepatocellular carcinoma (HCC) is the third leading cause of cancer-related death worldwide. Still, there are epidemiological and clinical data in Latin America. In Chile, this is the first study regarding HCC with a significant number of patients. This study aimed to obtain and analyze clinical and epidemiological data of Chilean patients with HCC. Materials and Methods: Multicenter study from 12 Chilean hospitals that have members of the Chilean Association of Hepatology as members of their staff. Clinical records from 2015-2021 were included. Kaplan-Meier survival curves and Cox regression analysis were obtained. Results: We obtained data from 856 patients with HCC from 12 different centers. Median age 67 years old; 58.7% male. Cirrhosis is present in 91.2% (779) of cases. Main risk factors reported: fatty liver 47.9%(410), alcohol 19.6% (68), viral 12.2%(104) and autoimmune 3.5% (30). The median MELD score was 11.7 (CI95% 11,4-12). 38% (322) were diagnosed during surveillance; this was associated with earlier BCLC stage (OR 2,6; CI95%1,9-3,4). BCLC stages at diagnosis were 0; 8,2%(69), A: 38,5%(326), B:29,9%(253), C: 15,4%(130) and D: 8,2%(69). The main initial treatments were TACE, ablation, resection, liver transplant and sorafenib in 27,4%(226), 19,3%(159), 11,4%(94), 8%(66) and 5,5%(45), respectively. 53,4%(452) pts were in Milan Criteria at diagnosis. 9,1%(78) patients got a liver transplant. Five-year survival was 24% (CI95%20-28). The main factors associated with survival are depicted in Figure 1. Conclusions: Fatty liver was remarkably the main risk factor reported for HCC in our Chilean cohort. This is a worrisome number since NAFLD is on the rise worldwide, and especially in Latin America. Surveillance is key for early detection. The liver function defined by Child-Pugh and HCC stage using BCLC staging is strongly associated with survival. Liver transplant is still a scarce treatment resource.

Intubating laryngeal mask airways can be used to provide continuous ventilation throughout intubation. This is a case of a morbidly obese (body mass index = 58) 65-year-old woman with T10 and T11 compression fractures. Optimal positioning for airway management was hindered by her unstable spine, minimal neck range of motion, and extreme pain with any movement. An intubating laryngeal mask airway was placed in the awake, topically anesthetized patient, and the laryngeal mask airway and endotracheal tube combination was left in place throughout surgery., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Objective: Recent evidence suggests that amyloid precursor protein (APP) is overexpressed in atherosclerosis-prone regions of mouse aorta. We therefore investigated in the present study whether APP has a role in the progression and composition of atherosclerotic plaques., Methods and Results: Apolipoprotein E-deficient (apoE(-/-)) mice were crossbred with animals lacking APP (APP(-/-)). After 16 weeks on a Western-type diet, apoE(-/-) and APP(-/-)/apoE(-/-) mice showed similar cholesterol levels. However, atherosclerotic plaque size was significantly reduced in the distal thoracic aorta (90% reduction) and abdominal aorta (75% reduction) of APP(-/-)/apoE(-/-) mice as compared to apoE(-/-). Plaques at the level of the aortic valves were not different in size, but showed a more stable phenotype in APP(-/-)/apoE(-/-) mice, as indicated by a reduced macrophage content, an increased amount of collagen and a thicker fibrous cap., Conclusion: Our findings provide evidence that lack of APP attenuates atherogenesis and leads to plaque stability., (Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.)

To study the effect of hypercholesterolemia on vascular smooth muscle cell (VSMC) function, atherosclerosis-prone but plaque-free endothelium-denuded aortic rings (width 2mm) from C57Bl6 Wild Type (WT) and apolipoprotein E-deficient (apoE(-/-)) mice (age 4 months) were mounted in a myograph and loaded with Fura-2 AM to simultaneously measure free Ca(2+) ([Ca(2+)](i)) and force development. In comparison with WT, apoE(-/-) mice displayed higher basal [Ca(2+)](i). Moreover, the time constant of the second phase of the biphasic high K(+)-induced [Ca(2+)](i) response was significantly increased in apoE(-/-) compared to WT mice. This phase was abolished by treatment with cyclopiazonic acid (CPA), depleting sarcoplasmic reticulum (SR). Further investigation of SR dependent [Ca(2+)](i) handling with CPA and caffeine revealed no alteration of maximal SERCA or ryanodine receptor function. Inositol (1,4,5)-triphosphate receptor (IP(3)R)-mediated [Ca(2+)](i) release was, however, significantly increased in apoE(-/-) mice compared to WT mice as established with phenylephrine and ATP. In Ca(2+)-free conditions the ATP-induced [Ca(2+)](i) was not altered. The ATP-induced store-operated Ca(2+) entry was, however, significantly increased in apoE(-/-) compared to WT mice. The results demonstrate that basal [Ca(2+)](i) levels and IP(3)R-mediated store-operated [Ca(2+)](i) release over the plasma membrane were elevated in hypercholesterolemic but plaque-free apoE(-/-) mice.

Defining the activity of an enzyme in a cell-based system is preferable to in vitro biochemical techniques requiring extensive purification. Chiang et al. use small molecules, RNA interference, and metabolomics to characterize a novel enzyme upregulated in cancer cells.

We investigated the participation of different tachykinin receptors in contractility of circular muscle strips of the mouse ileum using selective NK receptor agonists and antagonists. The NK1 receptor agonist septide (1-100 nM) induced dose-dependent contractions which were reduced by atropine and augmented by L-NNA. L-NNA increased and TTX consecutively reduced contractions to the NK2 receptor agonist beta-A-NKA (1-100 nM). Senktide, agonist of NK3 receptors, failed to induce contractions. NANC contractions to EFS were decreased after NK1 receptor blockade with RP67580. This inhibitory effect was more pronounced after additional blockade of NK2 and NK3 receptors. NK3 receptor antagonism alone reduced contractions at higher frequencies of stimulation. When the duration of the EFS stimulus was increased, the participation of all NK receptor subtypes became more evident. Our results suggest that excitatory NANC transmission in the circular muscle layer of the mouse ileum is mediated by tachykinins acting principally on NK1 receptors on cholinergic nerves and smooth muscle cells. Also NK2 receptors, located on smooth muscle cells and nitrergic neurons, and NK3 receptors on enteric neurons are involved.

Recent clinical evidence has indicated that the severity of atherosclerosis is correlated with the level of soluble ICAM-1 (sICAM-1). Nitric oxide (NO) donors are used to treat patients with stable angina pectoris, and the aim of this study was to investigate the short- and long-term effect of molsidomine on the level of this circulating biochemical marker of endothelial function. We included 172 patients and examined the effect of the NO donor treatment on angina related parameters and on sICAM-1 levels after a 4-week- and a 1-year treatment period. After 4 weeks, angina attacks and sublingual (s.l.) isosorbide dinitrate tablet (ISDN) consumption frequency was significantly (p<0.0001) reduced without altering sICAM-1 levels when compared to the baseline values. The anti-anginal effect of molsidomine 16 mg once a day (o.a.d.) was sustained (s.l. ISDN consumption) or improved (angina attacks frequency; p<0.002) during the following year and a significant decrease in sICAM-1 levels (p<0.0001) was observed. When the sICAM-1 changes during the 1-year treatment period were distributed in four categories (quartiles of the distribution), it was demonstrated that the decrease in s.l. ISDN consumption between the start and the end, was most pronounced in the group with the largest sICAM-1 decrease (fourth quartile of distribution; p=0.038). In conclusion, the reduction in the pro-inflammatory marker sICAM-1 after 1-year daily treatment with molsidomine may indicate that this NO donor besides its anti-anginal function, promotes a less activated state of the endothelium in patients with stable angina.

Previous studies demonstrated that macrophages are difficult to transfect. In the present study, we investigated whether J774A.1 macrophages can be efficiently transfected using nucleofector technology. Nucleofection of J774A.1 macrophages with mRNA resulted in transfection efficiencies up to 75% without cell death as compared to control pulsed macrophages. In contrast, introduction of DNA into J774A.1 cells caused apoptosis without expression of the gene of interest. Our results show that mRNA nucleofection is a new high-speed transfection method for macrophages.

Phagocyte recognition of cells undergoing apoptosis is a rapid, efficient way of removing unwanted cells from tissue. The uptake of apoptotic cells prevents the release of potentially toxic cell contents that might otherwise damage neighbouring cells and elicit an inflammatory response. The aim of this work was to evaluate a simple cell culture assay to study phagocytosis of cells undergoing apoptosis. Fluorescent negatively charged beads (1 microm) or fluorescently labelled apoptotic cells, derived from etoposide-treated human monocytes (U937), were co-incubated with J774 cells or human peripheral blood macrophages for 1 h. Flow cytometry (FCM) showed an efficient uptake of both beads and apoptotic bodies. Phagocytosis of apoptotic cells but not of beads was significantly inhibited when macrophages were pre-incubated with cytochalasin D, suggesting that an experimental system based on beads is not an appropriate model of phagocytosis of apoptotic cells.

Apoptosis or programmed cell death is a cellular suicide mechanism that frequently occurs in advanced human atherosclerotic plaques. Caspases, a family of cysteine proteases, have been identified as important effectors of the death machinery. In this study, we report strong caspase-2 immunoreactivity in foam cells of macrophage-origin around the necrotic core of advanced human atherosclerotic plaques. In contrast, smooth muscle cells (SMCs) and macrophages in the fibrous cap as well as endothelial cells, medial SMCs, and SMCs from mammary arteries are negative for caspase-2. Caspase-2-positive macrophages were isolated from human plaques by laser capture microdissection and were then analyzed by Western blotting. A single band of approximately 35 kd corresponding with the precursor of the short, anti-apoptotic isoform of caspase-2 (caspase-2S) could be identified. Treatment of human U937 macrophages with the DNA strand-breaking agents etoposide or camptothecin stimulated caspase-2S expression. Since atherosclerotic plaques contain a high number of DNA strand breaks, our results provide evidence for a survival factor in macrophage-derived foam cells of human atherosclerotic plaques that might be up-regulated in response to DNA damage.

In human atherosclerosis the development of a cell-poor lipid-rich core is an important feature of atheromatous plaque formation. The core is characterized by extracellular lipid deposition, cholesterol crystals and cell death and is situated in the deep layer of the plaque. The aim of the present study was to localize apoptotic cell death and cell replication in atherosclerotic plaques of cholesterol-fed rabbits in order to examine the hypothesis that core formation is a consequence of an imbalance between cell replication and apoptosis. New Zealand White male rabbits were fed a diet supplemented with 0.3% cholesterol for 16 (n = 5) and 27 weeks (n = 9). Cell replication and cell types were demonstrated by immunohistochemistry and apoptotic cell death was demonstrated by DNA in situ end-labeling (ISEL) and transmission electron microscopy. Quantification was done using a colour image analysis system. The plaques showed a clear distinction between a luminal layer composed of numerous lipid-rich foam cells of macrophage origin and a deep layer which was fibrous, containing extracellular lipid deposits and few smooth muscle cells. Cell replication (expressed as percentage of total number of nuclei) in the superficial layer was higher then in the deep layer at both 16 (5.1 +/- 1.8% vs. 1.2 +/- 0.8%) and 27 weeks (11.3 +/- 2.1% vs. 4.4 +/- 1.0%). This was also the case for the total number of nuclei per 50000 microns2 cross-sectional intimal area (numerical density): 235 +/- 13 vs. 147 +/- 7 at 16 weeks and 130 +/- 10 vs. 89 +/- 11 at 27 weeks. Apoptotic cell death (expressed as percentage of total number of nuclei) was low and there was no difference between the superficial and the deep layers of the plaques (0.8% +/- 0.2% vs. 0.4% +/- 0.2% at 16 weeks and 0.6 +/- 0.2% vs. 1.7% +/- 0.6% at 27 weeks). Our results indicate that the control of cell number in superficial vs. deep regions of the plaque is mainly a consequence of differences in cell replication. This may be due to a gradient of endothelial and plasma-derived growth factors. Cells can disappear by apoptosis, albeit at a relatively low level, throughout the lesion. This process may contribute to the pronounced cell loss in more advanced human atherosclerotic plaques, setting the base for plaque rupture.

To test the possible role of nitric oxide production in long-term portal vein ligation in the rat, where the hyperdynamic circulation was reported to be absent, in vivo experiments on isolated thoracic aortic rings from partial portal vein ligated or sham-operated rats were performed, 6 months postoperatively. The concentration-response curves to noradrenaline of both intact and endothelium-denuded rings from portal hypertensive rats were significantly shifted to the right as compared to those from sham-operated animals. In intact rings, addition of NG-nitro-L-arginine, a specific inhibitor of nitric oxide synthase, resulted in a significant shift of the curves to the left in sham-operated and portal vein ligated rats. In endothelium-denuded rings, addition of NG-nitro-L-arginine resulted in a significant shift of the curves to the left in portal vein ligated but not in sham-operated animals. After blockade of the nitric oxide biosynthesis with NG-nitro-L-arginine, the negative logarithm of the concentration of nonadrenaline causing half-maximal response did not significantly differ any more between portal vein ligated and sham-operated rats; in endothelium-denuded rings hyporeactivity to noradrenaline persisted in portal vein ligated rats. Only in the intact rings did NG-nitro-L-arginine significantly increase the maximal contractions. No differences were demonstrated in endothelium-dependent relaxations to acetylcholine between sham-operated and portal hypertensive animals. From these results, it can be concluded that in vitro aortic hyporeactivity to noradrenaline is still present in long-term portal vein ligated rats, and that it results at least partially from activation of the L-arginine: nitric oxide pathway in the aortic vascular wall.

Neutrophil-activating protein-2 (NAP-2), an NH2-terminally processed form of the platelet-release product beta thromboglobulin (beta TG), was purified to homogeneity from stimulated human blood leukocytes. In the presence of a vasodilator substance (PGE2, CGRP) picomolar (pmol/l) amounts of NAP-2 induced neutrophil accumulation and plasma leakage on intradermal injection in rabbit skin, whereas the longer forms, beta TG itself and connective tissue-activating peptide III (CTAP-III), had no such effect. NAP-2-induced increased in microvascular permeability was neutrophil dependent and fast in onset, with a half-life of 65 to 75 minutes, comparable to that previously reported for the structural-related neutrophil-activating protein-1/interleukin-8 (NAP-1/IL-8). However NAP-2 showed a lower potency in that more protein was needed to provoke skin reactivity. Nevertheless the finding that a platelet release product can elicit neutrophil-mediated inflammation further narrows the gap between thrombotic events and inflammatory disorders.

Granulocyte chemotactic protein/Interleukin-8 (GCP/IL-8), purified to homogeneity from endotoxin- or mitogen-stimulated human mononuclear cells, was injected intradermally into rabbits to evaluate the proinflammatory properties of this novel cytokine. In the presence of a vasodilator substance, pmol amounts of GCP/IL-8 induced neutrophil accumulation that was fast in onset, relatively short of duration (half life 60 to 70 minutes), and was associated with a parallel time course of plasma protein extravasation. GCP/IL-8-induced edema formation was found to be neutrophil dependent. These data provide evidence that GCP/IL-8 fulfills two important criteria for consideration as an inflammatory mediator. It is possible that endogenous GCP/IL-8, if produced locally by tissue macrophages, may contribute to the initiation of the inflammatory response to infection.