Morphologic Severity of Acute Pancreatitis on Imaging Is Independently Associated with Opioid Dose Requirements in Hospitalized Patients.

BACKGROUND: Prior studies have evaluated clinical characteristics associated with opioid dose requirements in hospitalized patients with acute pancreatitis (AP) but did not incorporate morphologic findings on CT imaging. AIMS: We sought to determine whether morphologic severity on imaging is independently associated with opioid dose requirements in AP. METHODS: Adult inpatients with a diagnosis of AP from 2006 to 2017 were reviewed. The highest modified CT severity index (MCTSI) score and the daily oral morphine equivalent (OME) for each patient over the first 7 days of hospitalization were used to grade the morphologic severity of AP and calculate mean OME per day(s) of treatment (MOME), respectively. Multiple regression analysis was used to evaluate the association of MOME with MCSTI. RESULTS: There were 249 patients with AP, of whom 196 underwent contrast-enhanced CT. The mean age was 46 ± 13.6 years, 57.9% were male, and 60% were black. The mean MOME for the patient cohort was 60 ± 52.8 mg/day. MCTSI (ß = 3.5 [95% CI 0.3, 6.7], p = 0.03), early hemoconcentration (ß = 21 [95% CI 4.6, 39], p = 0.01) and first episode of AP (ß = - 17 [95% CI - 32, - 2.7], p = 0.027) were independently associated with MOME. Among the 19 patients undergoing ≥ 2 CT scans, no significant differences in MOME were seen between those whose MCTSI score increased (n = 12) versus decreased/remained the same (n = 7). CONCLUSION: The morphologic severity of AP positively correlated with opioid dose requirements. No difference in opioid dose requirements were seen between those who did versus those who did not experience changes in their morphologic severity.

S-Nitrosation of ß-catenin and p120 catenin: a novel regulatory mechanism in endothelial hyperpermeability.

RATIONALE: Endothelial adherens junction proteins constitute an important element in the control of microvascular permeability. Platelet-activating factor (PAF) increases permeability to macromolecules via translocation of endothelial nitric oxide synthase (eNOS) to cytosol and stimulation of eNOS-derived nitric oxide signaling cascade. The mechanisms by which nitric oxide signaling regulates permeability at adherens junctions are still incompletely understood. OBJECTIVE: We explored the hypothesis that PAF stimulates hyperpermeability via S-nitrosation (SNO) of adherens junction proteins. METHODS AND RESULTS: We measured PAF-stimulated SNO of ß-catenin and p120-catenin (p120) in 3 cell lines: ECV-eNOSGFP, EAhy926 (derived from human umbilical vein), and postcapillary venular endothelial cells (derived from bovine heart endothelium) and in the mouse cremaster muscle in vivo. SNO correlated with diminished abundance of ß-catenin and p120 at the adherens junction and with hyperpermeability. Tumor necrosis factor-α increased nitric oxide production and caused similar increase in SNO as PAF. To ascertain the importance of eNOS subcellular location in this process, we used ECV-304 cells transfected with cytosolic eNOS (GFPeNOSG2A) and plasma membrane eNOS (GFPeNOSCAAX). PAF induced SNO of ß-catenin and p120 and significantly diminished association between these proteins in cells with cytosolic eNOS but not in cells wherein eNOS is anchored to the cell membrane. Inhibitors of nitric oxide production and of SNO blocked PAF-induced SNO and hyperpermeability, whereas inhibition of the cGMP pathway had no effect. Mass spectrometry analysis of purified p120 identified cysteine 579 as the main S-nitrosated residue in the region that putatively interacts with vascular endothelial-cadherin. CONCLUSIONS: Our results demonstrate that agonist-induced SNO contributes to junctional membrane protein changes that enhance endothelial permeability.

Assessing the negative impact of chlorantraniliprole, isoxaflutole, and simazine pesticides on phospholipid membrane models and tilapia gill tissues.

The indiscriminate and, very often, incorrect use of pesticides in Brazil, as well as in other countries, results in severe levels of environmental pollution and intoxication of human life. Herein, we studied plasma membrane models (monolayer and bilayer) of the phospholipid Dioleoyl-sn-glycerol-3-phosphocholine (DOPC) using Langmuir films, and large (LUVs) and giant (GUVs) unilamellar vesicles, to determine the effect of the pesticides chlorantraniliprole (CLTP), isoxaflutole (ISF), and simazine (SMZ), used in sugarcane. CLTP affects the lipid organization of the bioinspired models of DOPC π-A isotherms, while ISF and SMZ pesticides significantly affect the LUVs and GUVs. Furthermore, the in vivo study of the gill tissue in fish in the presence of pesticides (2.0 × 10-10 mol/L for CLTP, 8.3 × 10-9 mol/L for ISF, and SMZ at 9.9 × 10-9 mol/L) was performed using optical and fluorescence images. This investigation was motivated by the gill lipid membranes, which are vital for regulating transporter activity through transmembrane proteins, crucial for maintaining ionic balance in fish gills. In this way, the presence of phospholipids in gills offers a model for understanding their effects on fish health. Histological results show that exposure to CLTP, ISF, and SMZ may interfere with vital gill functions, leading to respiratory disorders and osmoregulation dysfunction. The results indicate that exposure to pesticides caused severe morphological alterations in fish, which could be correlated with their impact on the bioinspired membrane models. Moreover, the effect does not depend on the exposure period (24h and 96h), showing that animals exposed to pesticides for a short period suffer irreparable damage to gill tissue. In summary, we can conclude that the harm caused by pesticides, both in membrane models and in fish gills, occurs due to contamination of the aquatic system with pesticides. Therefore, water quality is vital for the preservation of ecosystems.

Endomucin marks quiescent long-term multi-lineage repopulating hematopoietic stem cells and is essential for their transendothelial migration.

Endomucin (EMCN) currently represents the only hematopoietic stem cell (HSC) marker expressed by both murine and human HSCs. Here, we report that EMCN+ long-term repopulating HSCs (LT-HSCs; CD150+CD48-LSK) have a higher long-term multi-lineage repopulating capacity compared to EMCN- LT-HSCs. Cell cycle analyses and transcriptional profiling demonstrated that EMCN+ LT-HSCs were more quiescent compared to EMCN- LT-HSCs. Emcn-/- and Emcn+/+ mice displayed comparable steady-state hematopoiesis, as well as frequencies, transcriptional programs, and long-term multi-lineage repopulating capacity of their LT-HSCs. Complementary functional analyses further revealed increased cell cycle entry upon treatment with 5-fluorouracil and reduced granulocyte colony-stimulating factor (GCSF) mobilization of Emcn-/- LT-HSCs, demonstrating that EMCN expression by LT-HSCs associates with quiescence in response to hematopoietic stress and is indispensable for effective LT-HSC mobilization. Transplantation of wild-type bone marrow cells into Emcn-/- or Emcn+/+ recipients demonstrated that EMCN is essential for endothelial cell-dependent maintenance/self-renewal of the LT-HSC pool and sustained blood cell production post-transplant.

Functional significance of cytosolic endothelial nitric-oxide synthase (eNOS): regulation of hyperpermeability.

Endothelial NOS (eNOS)-derived NO is a key factor in regulating microvascular permeability. We demonstrated previously that eNOS translocation from the plasma membrane to the cytosol is required for hyperpermeability. Herein, we tested the hypothesis that eNOS activation in the cytosol is necessary for agonist-induced hyperpermeability. To study the fundamental properties of endothelial cell monolayer permeability, we generated ECV-304 cells that stably express cDNA constructs targeting eNOS to the cytosol or plasma membrane. eNOS-transfected ECV-304 cells recapitulate the eNOS translocation and permeability properties of postcapillary venular endothelial cells (Sánchez, F. A., Rana, R., Kim, D. D., Iwahashi, T., Zheng, R., Lal, B. K., Gordon, D. M., Meininger, C. J., and Durán, W. N. (2009) Proc. Natl. Acad. Sci. U.S.A. 106, 6849-6853). We used platelet-activating factor (PAF) as a proinflammatory agonist. PAF activated eNOS by increasing phosphorylation of Ser-1177 and inducing dephosphorylation of Thr-495, increasing NO production, and elevating permeability to FITC-dextran 70 in monolayers of cells expressing wild-type and cytosolic eNOS. PAF failed to increase permeability to FITC-dextran 70 in monolayers of cells transfected with eNOS targeted to the plasma membrane. Interestingly, this occurred despite eNOS Ser-1177 phosphorylation and production of comparable amounts of NO. Our results demonstrate that the presence of eNOS in the cytosol is necessary for PAF-induced hyperpermeability. Our data provide new insights into the dynamics of eNOS and eNOS-derived NO in the process of inflammation.

Clinical impact of clonal hematopoiesis in patients with lymphoma undergoing ASCT: a national population-based cohort study.

Clonal hematopoiesis of indeterminate potential (CHIP) is suspected of being a risk factor for patients with cancer. This study aimed to assess the clinical consequences of CHIP in patients with lymphoma intended for high-dose chemotherapy and autologous stem-cell transplantation (ASCT) in a population-based setting. We identified 892 lymphoma patients who had undergone stem cell harvest at all transplant centers in Denmark. A total of 565 patients had an available harvest sample, which was analysed for CHIP by next-generation sequencing, and the median follow-up was 9.1 years. Of the patients who were intended for immediate ASCT, 25.5% (112/440) carried at least one CHIP mutation. In contrast to previous single-center studies CHIP was not associated with inferior overall survival (OS) in multivariate analyses. However, patients with mutations in genes of the DNA repair pathway (PPM1D, TP53, RAD21, BRCC3) had a significant inferior OS (HR after 1 year of follow-up 2.79, 95% confidence interval 1.71-4.56; p < 0.0001), which also was evident in multivariate analysis (p = 0.00067). These patients had also increased rates of therapy-related leukemia and admission to intensive care. Furthermore, in patients who did not undergo immediate ASCT, a significant inferior OS of individuals with DNA repair mutations was also identified (p = 0.003).

Androgen receptor CAG and GGN repeat polymorphisms influence performance in boys and girls.

BACKGROUND: Shorter CAG and GGN androgen receptor (AR) repeat polymorphisms are associated with stronger androgen signaling, and therefore, could influence lean mass and exercise performance during growth. METHODS: Physical fitness and body composition were measured by standardized procedures and the length of CAG and GGN repeats was determined by PCR and fragment analysis in 152 boys (11.5±2.6 years; Tanner ≤5) and 116 girls (10.1±3.2 years; Tanner ≤5). Individuals were grouped as CAG short (CAGS) if harboring repeat lengths of ≤21 and CAG long (CAGL) if CAG >21. Moreover, subjects were grouped as GGN short (GGNS) if harboring repeat lengths of ≤23 and GGN long (GGNL) if GGN>23. RESULTS: No significant differences in anthropometrics and body composition were observed between either CAGS and CAGL groups and GGNS and GGNL groups. Boys harboring CAGS completed the 300-meter test faster than their CAGL counterparts. Moreover, girls from the GGNL group showed a significant higher VO2max than those in the GGNS group. CONCLUSIONS: In summary, carrying a short allele of the androgen receptor CAG repeat polymorphism is associated to higher anaerobic performance in boys, whereas long alleles of androgen receptor GGN polymorphisms are associated to higher aerobic capacity in girls.

ANDROGEN RECEPTOR CAG AND GGN REPEAT POLYMORPHISMS AND BONE MASS IN BOYS AND GIRLS.

INTRODUCTION: the human androgen receptor (AR) gene possesses two trinucleotide polymorphic repeats, (CAG and GGN) that affect the amount of AR protein translated. In this study, we genotyped these polymorphic tracts in a representative sample of Caucasian children (Tanner ≤ 5), 152 boys (11.5 } 2.6 yrs) and 116 girls (10.1 } 3.2 yrs) from Spain and investigated their association with bone mass. METHODS: the length of CAG and GGN repeats was determined by PCR and fragment analysis. Body composition was assessed by dual energy x-ray absorptiometry (DXA). Individuals were grouped as CAG short (CAGS) if harboring repeat lengths of ≤ 21 and CAG long (CAGL) if CAG > 21. Moreover, subjects were grouped as GGN short (GGNS) if harboring repeat lengths of ≤ 23 and GGN long (GGNL) if GGN > 23. RESULTS: in boys, significant differences in height, body mass, whole body bone mineral density (BMD) and content (BMC), upper extremities BMC, lower extremities BMC, femoral neck BMD, Ward's triangle BMC and BMD and lumbar spine BMD were observed between CAGS and CAGL groups (P < 0.05). Thus, upper extremities BMD differed between GGNS and GGNL groups. After adjusting for confounding variables, only upper extremities BMD between GGNS and GGNL groups remained significant (P < 0.05). No differences were observed in girls in any measured site in relation to either CAG or GGN polymorphisms length. CONCLUSIONS: our results support the hypothesis that longer alleles of the AR CAG and GGN polymorphisms are associated with increased bone mass in prepubertal boys.

Introducción: el gen humano del receptor de androgenos (AR) posee dos repeticiones polimorficas de trinucleotidos (CAG y GGN) que afectan a la cantidad de proteina AR traducida. En este estudio, genotipamos esos tractos polimorficos en una muestra representativa de ninos caucasicos espanoles (Tanner ≤ 5), compuesta por 152 ninos (11.5 } 2.6 anos) y 116 ninas (10.1 } 3.2 anos) e investigamos su asociacion con la masa osea. Métodos: la longitud de las repeticiones CAG y GGN se determino mediante PCR y analisis de fragmentos. La composicion corporal se midio mediante absorciometria dual de rayos X (DXA). Los participantes fueron agrupados como CAG cortos (CAGS) si poseian una longitud de repeticiones ≤ 21 y CAG largos si esta era > 21. Ademas, los participantes se agruparon como GGN cortos (GGNS) si poseian una longitud de repeticiones ≤ 23 y GGN largos (GGNL) si esta era > 23. Resultados: en los ninos se encontraron diferencias en talla, peso corporal, densidad mineral osea (BMD) y contenido mineral oseo (BMC) del cuerpo entero, BMC de las extremidades superiores e inferiores, BMD del cuello del femur, BMC y BMD del triangulo de Ward's y BMD de la espina lumbar entre los grupos CAGS y CAGL (P < 0,05). Ademas, el BMD de las extremidades superiores fue significativamente diferente entre los grupos GGNS y GGNL. Tras ajustar por variables confusoras, la unica diferencia que se mantuvo significativa fue la del BMD en las extremidades superiores entre los grupos GGNS y GGNL (P < 0,05). No se observaron diferencias entre los grupos CAG y GGN y la masa osea en las ninas. Conclusiones: nuestros resultados apoyan la hipotesis de que los alelos largos de los polimorfismos CAG y GGN del AR estan asociados con una mayor masa osea en ninos prepuberes.

Androgen receptor CAG and GGN repeat polymorphisms and bone mass in boys and girls / Polimorfismos CAG y GGN del gen del receptor de andrógenos y masa ósea en niños y niñas

Introduction: the human androgen receptor (AR) gene possesses two trinucleotide polymorphic repeats, (CAG and GGN) that affect the amount of AR protein translated. In this study, we genotyped these polymorphic tracts in a representative sample of Caucasian children (Tanner ≤5), 152 boys (11.5±2.6 yrs) and 116 girls (10.1±3.2 yrs) from Spain and investigated their association with bone mass. Methods: the length of CAG and GGN repeats was determined by PCR and fragment analysis. Body composition was assessed by dual energy x-ray absorptiometry (DXA). Individuals were grouped as CAG short (CAGS) if harboring repeat lengths of ≤21 and CAG long (CAGL) if CAG >21. Moreover, subjects were grouped as GGN short (GGNS) if harboring repeat lengths of ≤23 and GGN long (GGNL) if GGN >23. Results: in boys, significant differences in height, body mass, whole body bone mineral density (BMD) and content (BMC), upper extremities BMC, lower extremities BMC, femoral neck BMD, Ward’s triangle BMC and BMD and lumbar spine BMD were observed between CAGS and CAGL groups (P<0.05). Thus, upper extremities BMD differed between GGNS and GGNL groups. After adjusting for confounding variables, only upper extremities BMD between GGNS and GGNL groups remained significant (P<0.05). No differences were observed in girls in any measured site in relation to either CAG or GGN polymorphisms length. Conclusions: our results support the hypothesis that longer alleles of the AR CAG and GGN polymorphisms are associated with increased bone mass in prepubertal boys (AU)

Introducción: el gen humano del receptor de andrógenos (AR) posee dos repeticiones polimórficas de trinucleótidos (CAG y GGN) que afectan a la cantidad de proteína AR traducida. En este estudio, genotipamos esos tractos polimórficos en una muestra representativa de niños caucásicos españoles (Tanner ≤5), compuesta por 152 niños (11.5±2.6 años) y 116 niñas (10.1±3.2 años) e investigamos su asociación con la masa ósea. Métodos: la longitud de las repeticiones CAG y GGN se determinó mediante PCR y análisis de fragmentos. La composición corporal se midió mediante absorciometría dual de rayos X (DXA). Los participantes fueron agrupados como CAG cortos (CAGS) si poseían una longitud de repeticiones ≤21 y CAG largos si esta era >21. Además, los participantes se agruparon como GGN cortos (GGNS) si poseían una longitud de repeticiones ≤23 y GGN largos (GGNL) si esta era >23. Resultados: en los niños se encontraron diferencias en talla, peso corporal, densidad mineral ósea (BMD) y contenido mineral óseo (BMC) del cuerpo entero, BMC de las extremidades superiores e inferiores, BMD del cuello del fémur, BMC y BMD del triángulo de Ward’s y BMD de la espina lumbar entre los grupos CAGS y CAGL (P<0,05). Además, el BMD de las extremidades superiores fue significativamente diferente entre los grupos GGNS y GGNL. Tras ajustar por variables confusoras, la única diferencia que se mantuvo significativa fue la del BMD en las extremidades superiores entre los grupos GGNS y GGNL (P<0,05). No se observaron diferencias entre los grupos CAG y GGN y la masa ósea en las niñas. Conclusiones: nuestros resultados apoyan la hipótesis de que los alelos largos de los polimorfismos CAG y GGN del AR están asociados con una mayor masa ósea en niños prepúberes (AU)

Functional role of gap junctions in cytokine-induced leukocyte adhesion to endothelium in vivo.

To assess the hypothesis that gap junctions (GJs) participate on leukocyte-endothelium interactions in the inflammatory response, we compared leukocyte adhesion and transmigration elicited by cytokine stimulation in the presence or absence of GJ blockers in the hamster cheek pouch and also in the cremaster muscle of wild-type (WT) and endothelium-specific connexin 43 (Cx43) null mice (Cx43e(-/-)). In the cheek pouch, topical tumor necrosis factor-alpha (TNF-alpha; 150 ng/ml, 15 min) caused a sustained increment in the number of leukocytes adhered to venular endothelium (LAV) and located at perivenular regions (LPV). Superfusion with the GJ blockers 18-alpha-glycyrrhetinic acid (AGA; 75 microM) or 18-beta-glycyrrhetinic acid (50 microM) abolished the TNF-alpha-induced increase in LAV and LPV; carbenoxolone (75 microM) or oleamide (100 microM) reduced LAV by 50 and 75%, respectively, and LPV to a lesser extent. None of these GJ blockers modified venular diameter, blood flow, or leukocyte rolling. In contrast, glycyrrhizin (75 microM), a non-GJ blocker analog of AGA, was devoid of effect. Interestingly, when AGA was removed 90 min after TNF-alpha stimulation, LAV started to rise at a similar rate as in control. Conversely, application of AGA 90 min after TNF-alpha reduced the number of previously adhered cells. In WT mice, intrascrotal injection of TNF-alpha (0.5 microg/0.3 ml) increased LAV (fourfold) and LPV (threefold) compared with saline-injected controls. In contrast to the observations in WT animals, TNF-alpha stimulation did not increase LAV or LPV in Cx43e(-/-) mice. These results demonstrate an important role for GJ communication in leukocyte adhesion and transmigration during acute inflammation in vivo and further suggest that endothelial Cx43 is key in these processes.

Conjunctivitis alérgica crónica asociada o no a la atopia respiratoria : posible presencia de la IgE total sérica en lágrimas / Chronic allergic conjunctivitis associated or not to respiratory atopy: posible presence of total serum IgE in tears

La IgE sérica puede filtrar a las lágrimas, por esto es posible que las enfermedades atópicas respiratorias pueden influir en los niveles locales en lágrimas en el curso de la conjuntivitis alérgica crónica (CAC) asociada a estas enfermedades, de ahí que se estudiaran 46 pacientes con CAC solitaria, 34 con CAC y rinitis, 27 con CAC, rinitis y asma, así como 59 sujetos controles sin antecedentes alérgicos personales ni familiares. Se determinó la IgE total en suero por punción venosa y en lágrimas por capilaridad, así como la correlación entre éstas. Se observó un incremento de la IgE a medida que se asociaban las enfermedades respiratorias a la CAC. La IgE sérica sólo tuvo significación en el grupo con CAC, rinitis y asma en relación con los controles y la IgE local en los 3 grupos. No hubo correlación entre la sérica y la de las lágrimas y hubo diferencias significativas entre los niveles locales de IgE entre la CAC solitaria con rinitis y asma, lo que sugiere una filtración de esta inmunoglobulina del suero a las lágrimas

Conjuntivitis alérgica crónica asociada o no a la atopia respiratoria : posible presencia de la IgE total sérica en lagrimas / Chronic allergic conjunctivitis associated or not to respiratory atopy : possible presence of total serum IgE in tears

La IgE sérica puede filtrar

Conjunctivitis alérgica crónica asociada o no a la atopia respiratoria : posible presencia de la IgE total sérica en lágrimas / Chronic allergic conjunctivitis associated or not to respiratory atopy: posible presence of total serum IgE in tears

La IgE sérica puede filtrar a las lágrimas, por esto es posible que las enfermedades atópicas respiratorias pueden influir en los niveles locales en lágrimas en el curso de la conjuntivitis alérgica crónica (CAC) asociada a estas enfermedades, de ahí que se estudiaran 46 pacientes con CAC solitaria, 34 con CAC y rinitis, 27 con CAC, rinitis y asma, así como 59 sujetos controles sin antecedentes alérgicos personales ni familiares. Se determinó la IgE total en suero por punción venosa y en lágrimas por capilaridad, así como la correlación entre éstas. Se observó un incremento de la IgE a medida que se asociaban las enfermedades respiratorias a la CAC. La IgE sérica sólo tuvo significación en el grupo con CAC, rinitis y asma en relación con los controles y la IgE local en los 3 grupos. No hubo correlación entre la sérica y la de las lágrimas y hubo diferencias significativas entre los niveles locales de IgE entre la CAC solitaria con rinitis y asma, lo que sugiere una filtración de esta inmunoglobulina del suero a las lágrimas