Prognostic impact of liver resection side in peri-hilar cholangiocarcinoma: A systematic review and meta-analysis.

BACKGROUND: Perihilar cholangiocarcinoma (pCCA) is a highly lethal hepatobiliary cancer. Radical resection offers the best chance for extended survival, but the efficacy of left-sided hepatectomy (LH) versus right-sided hepatectomy (RH) remains controversial. METHODS: A systematic review and meta-analysis of non-randomized cohort studies comparing LH and RH in patients with resectable pCCA was conducted. Subanalyses were performed based on year of publication, region, number of cases and Bismuth classification (BC) ≥ III. RESULTS: Nineteen studies involving 3838 patients were included, with 1779 (46 %) undergoing LH and 2059 (54 %) undergoing RH. LH was associated with increased overall survival (OS) in subgroup analysis of studies reporting hazard ratios (HR) (logHR 0.59; p = 0.04). LH showed higher rates of arterial resection (14 % vs. 1 %), transfusion (51 % vs. 41 %), operation time (MD 31.44 min), and bile leakage (21 % vs. 18 %), but lower rates of post-hepatectomy liver failure (9 % vs. 21 %) and 90-day mortality (8 % vs 16 %). Three-year disease-free survival rates increased in Western centers but decreased in Eastern centers. CONCLUSION: LH is linked to higher OS in this analysis but is a more demanding technique. Resection side decision should consider several factors, including future liver remnant, tumor location, vascular involvement, and surgical expertise.

Sfrp1 inhibits lung fibroblast invasion during transition to injury-induced myofibroblasts.

BACKGROUND: Fibroblast-to-myofibroblast conversion is a major driver of tissue remodelling in organ fibrosis. Distinct lineages of fibroblasts support homeostatic tissue niche functions, yet their specific activation states and phenotypic trajectories during injury and repair have remained unclear. METHODS: We combined spatial transcriptomics, multiplexed immunostainings, longitudinal single-cell RNA-sequencing and genetic lineage tracing to study fibroblast fates during mouse lung regeneration. Our findings were validated in idiopathic pulmonary fibrosis patient tissues in situ as well as in cell differentiation and invasion assays using patient lung fibroblasts. Cell differentiation and invasion assays established a function of SFRP1 in regulating human lung fibroblast invasion in response to transforming growth factor (TGF)ß1. MEASUREMENTS AND MAIN RESULTS: We discovered a transitional fibroblast state characterised by high Sfrp1 expression, derived from both Tcf21-Cre lineage positive and negative cells. Sfrp1 + cells appeared early after injury in peribronchiolar, adventitial and alveolar locations and preceded the emergence of myofibroblasts. We identified lineage-specific paracrine signals and inferred converging transcriptional trajectories towards Sfrp1 + transitional fibroblasts and Cthrc1 + myofibroblasts. TGFß1 downregulated SFRP1 in noninvasive transitional cells and induced their switch to an invasive CTHRC1+ myofibroblast identity. Finally, using loss-of-function studies we showed that SFRP1 modulates TGFß1-induced fibroblast invasion and RHOA pathway activity. CONCLUSIONS: Our study reveals the convergence of spatially and transcriptionally distinct fibroblast lineages into transcriptionally uniform myofibroblasts and identifies SFRP1 as a modulator of TGFß1-driven fibroblast phenotypes in fibrogenesis. These findings are relevant in the context of therapeutic interventions that aim at limiting or reversing fibroblast foci formation.

Percutaneous osteosynthesis of acetabular fractures with quadrilateral plate involvement using an infra-pectineal plate through a new paramedial approach: Description of the technique using cadaveric specimens.

Percutaneous osteosynthesis of acetabular fractures with quadrilateral plate involvement using an infra-pectineal plate through a new paramedial approach using cadaveric specimens. BACKGROUND: Intrapelvic approaches and infrapectineal plates have been used since the mid-nineties to solve Quadrilateral Plate osteosynthesis, with some problems in applying screws in the correct direction and difficulty in fracture reduction. We describe a minimally invasive paramedial approach and new ways to fix infrapectineal plates using one-step osteosynthesis (reduction and fixation). METHODS: Four transverse and four posterior hemitransverse acetabular fractures were reproduced using four fresh frozen cadavers. Acetabular osteosynthesis was performed using the paramedial approach. Sequential lasting time and reduction/stability quality were measured using analysis of variance (ANOVA) with Bonferroni Correction as the statistical method, registering iatrogenic injuries. RESULTS: Osteosynthesis was performed on seven acetabulae using infrapectineal horizontal plates for transverse fractures and vertical plates for posterior hemitransverse fractures. The duration of incision was 3:08 min and osteosynthesis was 55:12 min, with a total of 58:29 min. Median fracture displacement of 13.25 mm turned to a median of 0.01 mm once fracture osteosynthesis was performed with a p = 0.017. The peritoneum was injured twice and good osteosynthesis stability was observed. CONCLUSION: The paramedial approach is safe with direct access to key anatomical structures for acetabular osteosynthesis. Infrapectineal with reverse fixation plate osteosynthesis provides an excellent reduction rate and good stability once the implants act against displacement forces, making it possible to direct them freely. Further clinical and biomechanical trials are required to confirm our findings. We believe that there was an improvement of up to 60% in the result quality for some cases; however, this technique must be compared with other techniques. Evidence Level IV (Experimental Trial).

A different transcriptional landscape sheds light on Russian sturgeon (Acipenser gueldenstaedtii) mechanisms to cope with bacterial infection and chronic heat stress.

Sturgeons are chondrostean fish of high economic value and critically endangered due to anthropogenic activities, which has led to sturgeon aquaculture development. Russian sturgeon (Acipenser gueldenstaedtii), the second most important species reared for caviar, is successfully farmed in subtropical countries, including Uruguay. However, during the Uruguayan summer, sturgeons face intolerable warmer temperatures that weaken their defences and favour infections by opportunistic pathogens, increasing fish mortality and farm economic losses. Since innate immunity is paramount in fish, for which the liver plays a key role, we used deep RNA sequencing to analyse differentially expressed genes in the liver of Russian sturgeons exposed to chronic heat stress and challenged with Aeromonas hydrophila. We assembled 149.615 unigenes in the Russian sturgeon liver transcriptome and found that metabolism and immune defence pathways are among the top five biological processes taking place in the liver. Chronic heat stress provoked profound effects on liver biological functions, up-regulating genes related to protein folding, heat shock response and lipid and protein metabolism to meet energy demands for coping with heat stress. Besides, long-term exposure to heat stress led to cell damage triggering liver inflammation and diminishing liver ability to mount an innate response to A. hydrophila challenge. Accordingly, the reprogramming of liver metabolism over an extended period had detrimental effects on fish health, resulting in weight loss and mortality, with the latter increasing after A. hydrophila challenge. To our knowledge, this is the first transcriptomic study describing how chronic heat-stressed sturgeons respond to a bacterial challenge, suggesting that liver metabolism alterations have a negative impact on the innate anti-bacterial response.

Different response of Acipenser gueldenstaedtii CRP/SAP and SAA to bacterial challenge and chronic thermal stress sheds light on the innate immune system of sturgeons.

Sturgeons are chondrostean fish critically endangered due to anthropogenic loss and degradation of natural habitat and overfishing for meat and caviar production. Consequently, sturgeon aquaculture has extensively developed lately, being Russian sturgeon (Acipenser gueldenstaedtii) the second most important species reared for caviar production. However, Russian sturgeon aquaculture in subtropical countries, such as Uruguay, confronts difficulties because fish have to endure excessive summertime warm temperatures, which weaken their innate defences facilitating opportunistic infections. To address this problem, we look for identifying putative acute phase proteins (APPs), which might be robust serum biomarkers of both infection and chronic thermal stress, applied to monitoring Russian sturgeon health status in farms. We focused on the C-Reactive Protein/Serum Amyloid P (CRP/SAP) pentraxin since the pentraxin family includes well-known APPs, better characterised in mammals than fish. We identified A.gueldenstaedtii CRP/SAP (AgCRP/SAP), as a member of the universal CRP/SAP pentraxin sub-family, and studied AgCRP/SAP involvement in sturgeon response to bacterial challenge and chronic thermal stress, in comparison with A. gueldenstaedtii Serum Amyloid A (AgSAA), a previously described positive APP. Results showed that AgCRP/SAP is a constitutive serum component that remained constant upon Aeromonas hydrophila challenge and chronic thermal stress. Contrastingly, serum AgSAA was subjected to regulation by bacterial and thermal stress challenges, showing a 50-fold increase and 3-fold decline in serum levels, respectively. Overall, results highlight the potential value of AgSAA, but not of AgCRP/SAP, as a biomarker of bacterial infection and the need to continue searching for robust chronic thermal stress biomarkers in sturgeons.

Ocular Toxocariasis in Parasitology Consultation in Quindío, Colombia: Description of Cases and Contact Studies.

We report diagnosis, treatment and evolution of cases of ocular toxocariasis in specialized consultation in Quindío, Colombia. No cases were seen during the 2000-17 period, but five cases were confirmed from November 2017 to March 2019; two children resulted with definitive loss of vision on the affected eye. Studies in contacts found that 12 of 19 (63%) family members and 15 of 25 (60%) children <15 years of age living on the same street were positive for IgG Toxocara antibodies. Epidemiological studies are necessaries to establish the reasons for the increase in cases at this region.

Serum amyloid A is a positive acute phase protein in Russian sturgeon challenged with Aeromonas hydrophila.

The immune system of sturgeons, one of the most ancient and economically valuable fish worldwide, is poorly understood. The lack of molecular tools and data about infection biomarkers hinders the possibility to monitor sturgeon health during farming and detect infection outbreaks. To tackle this issue, we mined publicly available transcriptomic datasets and identified putative positive acute-phase proteins (APPs) of Russian sturgeons that could be induced by a bacterial infection and monitored using non-invasive methods. Teleost literature compelled us to focus on five promising candidates: hepcidin, a warm acclimation associated hemopexin, intelectin, serum amyloid A protein (SAA) and serotransferrin. Among them, SAA was the most upregulated protein at the mRNA level in the liver of sturgeons challenged with heat-inactivated or live Aeromonas hydrophila. To assess whether this upregulation yielded increasing SAA levels in circulation, we developed an in-house ELISA to quantify SAA levels in sturgeon serum. Circulating SAA rose upon bacterial challenge and positively correlated with hepatic saa expression. This is the first time serum SAA has been quantified in an Actinopterygii fish. Since APPs vary across different fish species, our work sheds light on sturgeon acute-phase response, revealing that SAA is a positive APP with potential value as infection biomarker.

Construcción de subjetividad trans a partir de la ocupación entendida como hacer, ser y llegar a ser / Construção da subjetividade trans a partir da ocupação compreendida como fazer, ser e chegar a ser / Construction of trans subjectivity from the occupation understood as doing, being and becoming

Resumen Introducción Esta investigación indaga la influencia del sistema binario de género en la construcción del sujeto trans desde la perspectiva del Hacer y las potencialidades de Llegar a Ser, en el contexto sociocultural y político chileno, centrándose en las experiencias de los participantes mediante el rescate de la historia ocupacional. Objetivos Analizar la construcción del sujeto trans a través del hacer, ser y llegar a ser en base a sus narrativas y en relación a la estructura social chilena en la cual se desenvuelven, visualizando la influencia en sus estados de salud. Metodo Desde una perspectiva cualitativa con enfoque de conocimientos situados, se utiliza la metodología de producciones narrativas y técnicas discontinuas a múltiples voces. La información fue recolectada mediante entrevistas individuales y un grupo focal. Resultados El proceso de tránsito se presenta como un continuo ocupacional, identificando la performatividad del género mediante la ocupación, así como las transformaciones corporales en la construcción de subjetividad. Además se identifica un contexto sociocultural y político que vulnera derechos, lo cual impacta en la salud y el involucramiento en ocupaciones colectivas. Conclusiones La ocupación se presenta como un elemento reproductor del sistema binario de sexo-género, no obstante, también puede ser entendida como una herramienta de emancipación en la medida que permite transgredirla. En este sentido, se interpela a la Terapia Ocupacional a promover ocupaciones colectivas que potencien la capacidad de agencia de sujetos abyectos, determinando estados de salud favorables.

Resumo Introdução Esta pesquisa investiga a influência do sistema binário de gênero na construção do sujeito trans desde a perspectiva do fazer e as potencialidades do tornar-se a ser, no contexto sociocultural e político chileno, enfocando as experiências dos participantes através do resgate de História Ocupacional. Objetivos Analisar a construção do sujeito trans através do fazer, ser e tornar-se, com base em suas narrativas e em relação à estrutura social chilena em que atuam, visualizando a influência em seus estados de saúde. Método Em uma perspectiva qualitativa, com foco no conhecimento situado, foi utilizada a metodologia de produção de narrativa e técnicas descontínuas com múltiplas vozes. As informações foram coletadas por meio de entrevistas individuais e um grupo focal. Resultados O processo de trânsito é apresentado como um continuum ocupacional, identificando a performatividade de gênero através da ocupação, bem como transformações corporais na construção da subjetividade. Além disso, é identificado um contexto sociocultural e político que viola os direitos, com impacto na saúde e no envolvimento em ocupações coletivas. Conclusões A ocupação é apresentada como um elemento reprodutor do sistema binário sexo-gênero, no entanto, também pode ser entendida como uma ferramenta de emancipação, na medida em que permite transgredi-la. Nesse sentido, interpela-se a Terapia Ocupacional a promover ocupações coletivas que aumentem a capacidade de agência de sujeitos abjetos, determinando estados de saúde favoráveis.

Abstract Introduction This research inquires the influence of the binary gender system on the construction of the trans subject from the perspective of Doing and the potential of becoming, in the Chilean socio-cultural and political context, focusing on the experiences of the participants through the rescue of history occupational. Objectives Analyze the construction of the trans subject through doing, being and becoming based on their narratives, the Chilean social structure in which they develop, visualizing the influence on their health states. Method From a qualitative perspective with focus on situated knowledge, the methodology of narrative productions and discontinuous techniques is used to multiple voices. The information was collected through individual interviews and a focus group. Results The transit process is presented as an occupational continuum, identifying the performativization of gender through occupation, such as bodily transformations, in the construction of subjectivity. In addition, a sociocultural and political context that violates rights is identified, which impacts on health and participation in collective occupations. Conclusions Occupation is presented as a reproductive element of the binary sex-gender system, however, it can also be understood as a tool of emancipation insofar as it transgresses it. In this sense, Occupational Therapy is called upon to promote collective occupations that enhance the capacity of agency of abject subjects, determining favorable health states.

CX3CR1-fractalkine axis drives kinetic changes of monocytes in fibrotic interstitial lung diseases.

Circulating immune cell populations have been shown to contribute to interstitial lung disease (ILD). In this study, we analysed circulating and lung resident monocyte populations, and assessed their phenotype and recruitment from the blood to the lung in ILD. Flow cytometry analysis of blood samples for quantifying circulating monocytes was performed in 105 subjects: 83 with ILD (n=36, n=28 and n=19 for nonspecific interstitial pneumonia, hypersensitivity pneumonitis and connective-tissue disease-associated ILD, respectively), as well as 22 controls. Monocyte localisation and abundance were assessed using immunofluorescence and flow cytometry of lung tissue. Monocyte populations were cultured either alone or with endothelial cells to assess fractalkine-dependent transmigration pattern. We show that circulating classical monocytes (CM) were increased in ILD compared with controls, while nonclassical monocytes (NCM) were decreased. CM abundance correlated inversely with lung function, while NCM abundance correlated positively. Both CCL2 and CX3CL1 concentrations were increased in plasma and lungs of ILD patients. Fractalkine co-localised with ciliated bronchial epithelial cells, thereby creating a chemoattractant gradient towards the lung. Fractalkine enhanced endothelial transmigration of NCM in ILD samples only. Immunofluorescence, as well as flow cytometry, showed an increased presence of NCM in fibrotic niches in ILD lungs. Moreover, NCM in the ILD lungs expressed increased CX3CR1, M2-like and phagocytic markers. Taken together, our data support that in ILD, fractalkine drives the migration of CX3CR1+ NCM to the lungs, thereby perpetuating the local fibrotic process.

Antigen B from Echinococcus granulosus is a novel ligand for C-reactive protein.

Antigen B (EgAgB) is a phosphatidylcholine (PC)-rich lipoprotein of Echinococcus granulosus s.l. larva, potentially capable of modulating the activation of various myeloid cells, including macrophages. As C-reactive protein (CRP) can act as an innate receptor with ability to bind the phosphocholine moiety of PC in lipoproteins, we investigated whether EgAgB and CRP could interact during cystic echinococcosis infection (CE), and how CRP binding could affect the modulation activities exerted by EgAgB on macrophages. To that end, we firstly investigated the occurrence of CRP induction during human CE. We found that 61% of CE patients, but none of healthy donors, exhibited serum CRP levels higher than 10 mg/mL, suggesting that CRP can be induced during the chronic phase of CE. Furthermore, human CRP was capable of binding specifically to EgAgB with high affinity (0.6 ± 0.1 nM); this binding was Ca2+ -dependent and involved the phosphocholine moiety of PC, but not EgAgB8/1, EgAgB8/2 or EgAgB8/3 apolipoproteins. Finally, CRP presence altered the modulation exerted by EgAgB on the cytokine response of LPS-activated macrophages. Overall, our results suggest that CRP presence during CE may contribute to a complex scenario of interactions between EgAgB and myeloid cells, influencing the cytokine response induced during macrophage activation.

Instrucción entre pares como metodología a utilizar en enfermería / Peer instruction as a methodology to be used in nursing

Este trabajo presenta resultados de una revisión de la literatura acerca de la implementación de la metodología interactiva de enseñanza Peer Instruction (PI). Se pretende con esta búsqueda responder a las siguientes preguntas: ¿En qué contextos de enseñanza se ha investigado el Peer Instruction (PI)? ¿Qué impactos ha producido el PI en el aprendizaje de los estudiantes? ¿Cuáles son los resultados de la implementación del PI en relación a las actitudes de los estudiantes y profesores frente al uso de la metodología? Se realizó la búsqueda en las bases de datos Pubmed, Web of Science, EBSCO y Google Scholar. Los resultados de la literatura apuntan que gran parte de las publicaciones fueron conducidas en universidades norteamericanas, mayoritariamente enfocadas a la disciplina de Física11,16-18 , pero también las ciencias médicas3-6,10 y matemáticas7-9 . La adopción del PI presenta impactos positivos en el aprendizaje conceptual de los estudiantes, en la capacidad de resolución de problemas y en el desempeño académico. Desarrolla sentimientos positivos relacionados con el aprendizaje de los contenidos y la metodología. Los profesores realizan modificaciones al implementar el PI integrándolo con otras metodologías, demostrando su flexibilidad. En enfermería se ha utilizado escasamente el PI, siendo mezclado con otras metodologías en donde el estudiante es centro del aprendizaje; peer teaching and peer learning, por ejemplo; con buenos resultados en el logro de los resultados de aprendizajes de los estudiantes, dando un puntapié inicial a seguir aplicando e investigando el impacto de esta metodología en los estudiantes de enfermería.

This paper presents the results of a review of the literature about the implementation of the Peer Instruction interactive teaching methodology (PI). The aim of this search is to answer the following questions: In what teaching contexts has Peer Instruction (PI) been investigated? What impacts has IP produced on student learning? What are the results of the implementation of the IP in relation to the attitudes of the students and professors in front of the use of the methodology? The search was performed in Pubmed, Web of Science, EBSCO and Google Scholar databases. The results of the literature indicate that a large part of the publications were conducted in North American universities, mostly focused on the discipline of Physics, but also the medical sciences,and mathematics. The adoption of the PI has positive impacts on students' conceptual learning, problem-solving ability and academic performance. Develop positive feelings related to the learning of content and methodology. The professors make modifications when implementing the PI integrating it with other methodologies, demonstrating their flexibility. In nursing, the PI has been used sparingly, being mixed with other methodologies where the student is the center of learning; peer teaching and peer learning, for example; with good results in the achievement of student learning outcomes, giving an initial kick to continue applying and investigating the impact of this methodology on nursing students.

Russian sturgeon cultured in a subtropical climate shows weaken innate defences and a chronic stress response.

Russian sturgeon (Acipenser gueldenstaedtii) has been successfully farmed in Uruguay for the past ten years. However, during the Uruguayan summer fish endure high water temperatures and increased bacterial infections that threaten aquaculture. Our understanding of sturgeon's immune system and its interplay with environmental factors like temperature is almost unknown. This study analysed the way in which seasonal variations affect enzymatic blood components of Russian sturgeon's innate defences, including the serum alternative complement pathway (ACP), ceruloplasmin (Cp) and lysozyme activities. Results showed that summertime conditions in the farm altered these defences in different ways, inducing a significant decrease in ACP and Cp, and an increase in lysozyme. In addition, serum levels of total protein and cortisol decreased in summer, suggesting a chronic stress response was induced in parallel. Subsequently, we analysed whether the increase in water river temperature during summer could account for the observed results. To that end, we acclimated juvenile sturgeons to mild (18 °C) or warm (24 °C) temperatures for 37 days. Like in summer, sturgeons exposed to 24 °C showed lower levels of serum ACP, Cp and total proteins, together with a progressive decrease in body weight and increased fish mortality. Administration of an immunostimulant containing Se and Zn slightly reverted the temperature-induced effects on sturgeon's defences. Altogether, our study provides novel data on various physiological parameters of the Russian sturgeon and highlights the impact warm temperature has on stress and innate immunity in this chondrostean fish.

Epidermolysis bullosa simplex with muscular dystrophy associated with PLEC deletion mutation.

Epidermolysis bullosa simplex with muscular dystrophy (EBS-MD; OMIM #226670) is an autosomal recessive disorder characterized by neonatal blistering and later-onset muscle weakness.

Echinococcus granulosus Antigen B binds to monocytes and macrophages modulating cell response to inflammation.

BACKGROUND: Antigen B (EgAgB) is an abundant lipoprotein released by the larva of the cestode Echinococcus granulosus into the host tissues. Its protein moiety belongs to the cestode-specific family known as hydrophobic ligand binding protein (HLBP), and is encoded by five gene subfamilies (EgAgB8/1-EgAgB8/5). The functions of EgAgB in parasite biology remain unclear. It may play a role in the parasite's lipid metabolism since it carries host lipids that E. granulosus is unable to synthesise. On the other hand, there is evidence supporting immuno-modulating activities in EgAgB, particularly on innate immune cells. Both hypothetical functions might involve EgAgB interactions with monocytes and macrophages, which have not been formally analysed yet. METHODS: EgAgB binding to monocytes and macrophages was studied by flow cytometry using inflammation-recruited peritoneal cells and the THP-1 cell line. Involvement of the protein and phospholipid moieties in EgAgB binding to cells was analysed employing lipid-free recombinant EgAgB subunits and phospholipase D treated-EgAgB (lacking the polar head of phospholipids). Competition binding assays with plasma lipoproteins and ligands for lipoprotein receptors were performed to gain information about the putative EgAgB receptor(s) in these cells. Arginase-I induction and PMA/LPS-triggered IL-1ß, TNF-α and IL-10 secretion were examined to investigate the outcome of EgAgB binding on macrophage response. RESULTS: Monocytes and macrophages bound native EgAgB specifically; this binding was also found with lipid-free rEgAgB8/1 and rEgAgB8/3, but not rEgAgB8/2 subunits. EgAgB phospholipase D-treatment, but not the competition with phospholipid vesicles, caused a strong inhibition of EgAgB binding activity, suggesting an indirect contribution of phospholipids to EgAgB-cell interaction. Furthermore, competition binding assays indicated that this interaction may involve receptors with affinity for plasma lipoproteins. At functional level, the exposure of macrophages to EgAgB induced a very modest arginase-I response and inhibited PMA/LPS-mediated IL-1ß and TNF-α secretion in an IL-10-independent manner. CONCLUSION: EgAgB and, particularly its predominant EgAgB8/1 apolipoprotein, are potential ligands for monocyte and macrophage receptors. These receptors may also be involved in plasma lipoprotein recognition and induce an anti-inflammatory phenotype in macrophages upon recognition of EgAgB.

Lipid-free antigen B subunits from echinococcus granulosus: oligomerization, ligand binding, and membrane interaction properties.

BACKGROUND: The hydatid disease parasite Echinococcus granulosus has a restricted lipid metabolism, and needs to harvest essential lipids from the host. Antigen B (EgAgB), an abundant lipoprotein of the larval stage (hydatid cyst), is thought to be important in lipid storage and transport. It contains a wide variety of lipid classes, from highly hydrophobic compounds to phospholipids. Its protein component belongs to the cestode-specific Hydrophobic Ligand Binding Protein family, which includes five 8-kDa isoforms encoded by a multigene family (EgAgB1-EgAgB5). How lipid and protein components are assembled into EgAgB particles remains unknown. EgAgB apolipoproteins self-associate into large oligomers, but the functional contribution of lipids to oligomerization is uncertain. Furthermore, binding of fatty acids to some EgAgB subunits has been reported, but their ability to bind other lipids and transfer them to acceptor membranes has not been studied. METHODOLOGY/PRINCIPAL FINDINGS: Lipid-free EgAgB subunits obtained by reverse-phase HPLC were used to analyse their oligomerization, ligand binding and membrane interaction properties. Size exclusion chromatography and cross-linking experiments showed that EgAgB8/2 and EgAgB8/3 can self-associate, suggesting that lipids are not required for oligomerization. Furthermore, using fluorescent probes, both subunits were found to bind fatty acids, but not cholesterol analogues. Analysis of fatty acid transfer to phospholipid vesicles demonstrated that EgAgB8/2 and EgAgB8/3 are potentially capable of transferring fatty acids to membranes, and that the efficiency of transfer is dependent on the surface charge of the vesicles. CONCLUSIONS/SIGNIFICANCE: We show that EgAgB apolipoproteins can oligomerize in the absence of lipids, and can bind and transfer fatty acids to phospholipid membranes. Since imported fatty acids are essential for Echinococcus granulosus, these findings provide a mechanism whereby EgAgB could engage in lipid acquisition and/or transport between parasite tissues. These results may therefore indicate vulnerabilities open to targeting by new types of drugs for hydatidosis therapy.

Echinococcus granulosus antigen B: a Hydrophobic Ligand Binding Protein at the host-parasite interface.

Lipids are mainly solubilized by various families of lipid binding proteins which participate in their transport between tissues as well as cell compartments. Among these families, Hydrophobic Ligand Binding Proteins (HLBPs) deserve special consideration since they comprise intracellular and extracellular members, are able to bind a variety of fatty acids, retinoids and some sterols, and are present exclusively in cestodes. Since these parasites have lost catabolic and biosynthetic pathways for fatty acids and cholesterol, HLBPs are likely relevant for lipid uptake and transportation between parasite and host cells. Echinococcus granulosus antigen B (EgAgB) is a lipoprotein belonging to the HLBP family, which is very abundant in the larval stage of this parasite. Herein, we review the literature on EgAgB composition, structural organization and biological properties, and propose an integrated scenario in which this parasite HLBP contributes to adaptation to mammalian hosts by meeting both metabolic and immunomodulatory parasite demands.

Inhibition of bacillus Calmette-Guérin-induced nitric oxide in bladder tumor cells may improve BCG treatment.

Bacillus Calmette-Guérin (BCG) is considered to be one of the most effective treatments for superficial and in situ bladder cancer. However, either failure to respond initially or relapse within the first 5 years of treatment has been observed in some patients. As nitric oxide (NO) has been detected in the bladder of BCG-treated patients, we analyzed the role of endogenous NO generated after BCG treatments on human (T24) and murine (MB49 and MBT2) bladder tumor cells in the viability of tumor and immune cells, both in vitro and in vivo. In vitro inhibition of cancer cells after BCG treatment was evaluated by cell titer assay. NO production was determined as nitrite by Griess reagent. The death of immunocytes was evaluated by 51Cr release. Tumor histology with hematoxylin and eosin and Masson's trichrome staining was performed. BCG induced a direct inhibition of tumor cell growth in vitro, independently of NO levels. Besides, BCG-mediated NO production by tumor cells induced the death of spleen and peritoneal cells in syngeneic mice. The in vivo inhibition of NO synthase (NOS) activity by NG-nitro-L-arginine methyl ester in combination with BCG, improved tumor regression by generating a healing tissue. The increase of NO generated after BCG administration may induce the death of immunocytes. The in vivo inhibition of NO ameliorated immunotherapy with BCG by additional tumor growth inhibition. Our results suggested the possibility that the final outcome of patients with bladder tumors may improve by modulating NOS activity concomitantly with BCG therapy.