A new score for airway assessment using clinical and ultrasound parameters.

Background: Over the last few years, ultrasonography has been introduced as the fifth pillar to patient's bedside physical examination. Clinical assessments aim to screen and look for airway difficulties to predict difficult intubations, but none have demonstrated a significant predictive capacity. Recent systematic reviews have established a correlation between ultrasound imaging and difficult direct laryngoscopy. The primary objective of this study was to determine whether the utilization of ultrasonography to examine the upper airway could accurately predict difficult direct laryngoscopy. Methods: This is a prospective observational study including 102 adult patients that required general anesthesia for elective surgery. Preoperatively, clinical airway assessments were performed. Data such as Mallampati-Samsoon grade (MS), upper lip bite test (ULBT), thyromental (TMD) and sternomental distance (SMD), cervical circumference (CC) and the Arné risk index were collected. Ultrasound evaluation was taken at five different levels in two planes, parasagittal and transverse. Therefore, the following measurements were registered: distance from skin to hyoid bone (DSHB), distance from skin to thyrohyoid membrane (DSTHM), distance from skin to epiglottis (DSE), distance from skin to thyroid cartilage (DSTC) and distance from hyoid bone and thyroid cartilage (DHBTC). Patients were divided into two groups based on the difficulty to perform direct laryngoscopy, according to Cormack-Lehane (C-L) classification. Grades I and II were classified as easy laryngoscopy and grades III or IV as difficult. Logistic regression models and the Receiver Operating Characteristic (ROC) curve was employed to determine the diagnostic precision of ultrasound measurements to distinguish difficult laryngoscopy (DL). Results: The following risk score for DL was obtained, DSTHM ≥ 1.60 cm (2 points), DSTC ≥ 0.78 cm (3 points) and gender (2 points for males). The score can range from 0 to 7 points, and showed and AUC (95% CI) of 0.84 (0.74-0.95). A score of 5 points or higher indicates a 34-fold increase in the risk of finding DL (p = 0.0010), sensitivity of 91.67, specificity of 75.56, positive predictive value of 33.33, and negative predictive value of 98.55. Conclusion: The use of ultrasonography combined with classic clinical screening tests are useful tools to predict difficult direct laryngoscopy.

Delayed referral for liver transplant evaluation worsens outcomes in chronic liver disease patients requiring inpatient transplant evaluation.

BACKGROUND: Indications to refer patients with cirrhosis for liver transplant evaluation (LTE) include hepatic decompensation or a model for end stage liver disease (MELD-Na) score ≥ 15. Few studies have evaluated how delaying referral beyond these criteria affects patient outcomes. AIM: To evaluate clinical characteristics of patients undergoing inpatient LTE and to assess the effects of delayed LTE on patient outcomes (death, transplantation). METHODS: This is a single center retrospective cohort study assessing all patients undergoing inpatient LTE (n = 159) at a large quaternary care and liver transplant center between 10/23/2017-7/31/2021. Delayed referral was defined as having prior indication (decompensation, MELD-Na ≥ 15) for LTE without referral. Early referral was defined as referrals made within 3 mo of having an indication based on practice guidelines. Logistic regression and Cox Hazard Regression were used to evaluate the relationship between delayed referral and patient outcomes. RESULTS: Many patients who require expedited inpatient LTE had delayed referrals. Misconceptions regarding transplant candidacy were a leading cause of delayed referral. Ultimately, delayed referrals negatively affected overall patient outcome and an independent predictor of both death and not receiving a transplant. Delayed referral was associated with a 2.5 hazard risk of death. CONCLUSION: Beyond initial access to an liver transplant (LT) center, delaying LTE increases risk of death and reduces risk of LT in patients with chronic liver disease. There is substantial opportunity to increase the percentage of patients undergoing LTE when first clinically indicated. It is crucial for providers to remain informed about the latest guidelines on liver transplant candidacy and the transplant referral process.

Trends and Variation in the Gap Between Current and Anticipated Life Satisfaction in the United States, 2008-2020.

Objectives. To describe national- and county-level trends and variation in a novel measure of hope. Methods. Using data from the Gallup National Health and Well-Being Index (n = 2 766 728), we summarized the difference between anticipated life satisfaction (ALS) and current life satisfaction (CLS), measured by the Cantril Self-Anchoring Scale, for each year from 2008 to 2020 and by county over two 5-year periods in the United States. Results. Across all years, there was a significant positive trend in the difference between ALS and CLS for the nation (P = .024), which remained positive but not significant when we excluded 2020. Maintenance of ALS with a decrease in CLS drove the 2020 increase. From 2008-2012 to 2013-2017, 14.5% of counties with 300 or more responses (n = 599) experienced an increase in the difference of more than 1 SD, whereas 13.9% experienced a more than 1 SD decrease. Fifty-two counties experienced decreases in ALS and CLS. Conclusions. Responding to trends in the gap between ALS and CLS at national and local levels is essential for the collective well-being of our nation, especially as we navigate and emerge from crisis.

Trends and geographical variation in population thriving, struggling and suffering across the USA, 2008-2017: a retrospective repeated cross-sectional study.

OBJECTIVES: Well-being is a holistic, positively framed conception of health, integrating physical, emotional, social, financial, community and spiritual aspects of life. High well-being is an intrinsically worthy goal for individuals, communities and nations. Multiple measures of well-being exist, yet we lack information to identify benchmarks, geographical disparities and targets for intervention to improve population life evaluation in the USA. DESIGN: Using data from the Gallup National Health and Well-Being Index, we conducted retrospective analyses of a series of cross-sectional samples. SETTING/PARTICIPANTS: We summarised select well-being outcomes nationally for each year, and by county (n=599) over two time periods, 2008-2012 and 2013-2017. MAIN OUTCOME MEASURES: We report percentages of people thriving, struggling and suffering using the Cantril Self-Anchoring Scale, percentages reporting high or low current life satisfaction, percentages reporting high or low future life optimism, and changes in these percentages over time. RESULTS: Nationally, the percentage of people that report thriving increased from 48.9% in 2008 to 56.3% in 2017 (p<0.05). The percentage suffering was not significantly different over time, ranging from 4.4% to 3.2%. In 2013-2017, counties with the highest life evaluation had a mean 63.6% thriving and 2.3% suffering while counties with the lowest life evaluation had a mean 49.5% thriving and 6.5% suffering, with counties experiencing up to 10% suffering, threefold the national average. Changes in county-level life evaluation also varied. While counties with the greatest improvements experienced 10%-15% increase in the absolute percentage thriving or 3%-5% decrease in absolute percentage suffering, most counties experienced no change and some experienced declines in life evaluation. CONCLUSIONS: The percentage of the US population thriving increased from 2008 to 2017 while the percentage suffering remained unchanged. Marked geographical variation exists indicating priority areas for intervention.

Predictors of close faculty-student relationships and mentorship in higher education: findings from the Gallup-Purdue Index.

College students' supportive relationships with mentors-professors, advisors, and other caring adults to whom students turn as they develop their interests and career paths-are critical to their development and academic success. The current study sought to explore factors that promote or impede the formation of positive mentor-student relationships during college using a large, nationally representative sample of 5,684 college graduates from the Gallup-Purdue Index. Linear regression models revealed that first-generation college students, as well as students attending larger institutions, rated faculty and other college staff as less caring and supportive, and were less able to identify a supportive mentoring relationship during college. Greater engagement at college, including participation in faculty research, academic internships, long-term projects, and extracurricular clubs or activities, was associated with stronger perceptions of faculty support and mentorship while in college. Interestingly, demographic characteristics moderated the effects of some extracurricular activities on students' experiences. For example, participants with more student loans showed a stronger positive association between participation in long-term academic projects and perceptions of faculty support, relative to students with few loans. These findings have important implications for policies designed to foster sustained and meaningful faculty-student relationships for all students, including those traditionally marginalized on college campuses.

Use, Yield, and Risk of Device-assisted Enteroscopy in the United States: Results From a Large Retrospective Multicenter Cohort.

INTRODUCTION: Since 2001, device-assisted enteroscopy (DAE) has revolutionized the diagnostic and therapeutic capabilities for managing small bowel pathology. Though commonly performed, there have been no recent large studies to assess the use, yield, and risks of DAE and none that include all 3 DAE modalities. We hypothesized that DAE is safe with high diagnostic and therapeutic yields achieved within reasonable procedure duration and here we present a large retrospective multicenter US study evaluating the use, yield, and complications of DAE. METHODS: After obtaining institutional review board approval, electronic records were used to identify all DAE's performed for luminal small bowel evaluation in adult patients at 4 US referral centers (Duke University Medical Center, New York University Langone Medical Center, Louisiana State University Health Sciences Center, and University of Massachusetts Medical Center) from January 1, 2014 to January 1, 2019. Electronic medical records were reviewed to collect and analyze a variety of procedure-related outcomes. Using the data pooled across centers, descriptive statistics were generated for the patient and procedure-related characteristics and outcomes; relationships between characteristics and outcomes were explored. RESULTS: A total of 1787 DAE's were performed over this 5-year period (392 at Duke University Medical Center, 887 at Louisiana State University Health Sciences Center, 312 at New York University Langone Medical Center, and 195 at University of Massachusetts Medical Center). Of these, there were 1017 (57%) double-balloon, 391 (29%) single-balloon, and 378 (21%) spiral enteroscopies. The mean age of patients undergoing DAE was 66 years and 53% of examinations were performed on women; 18% of patients in the cohort underwent >1 DAE over this time span. A total of 53% of examinations were performed for suspected small bowel bleeding, 31% were directly guided by video capsule endoscopy findings and 8% were performed for abnormal imaging. A total of 85% of examinations used an antegrade approach and DAE took a mean of 45 minutes to complete; 76% of examinations revealed abnormal findings, with vascular, inflammatory, and neoplastic findings seen in 49%, 17%, and 15% of the cohort, respectively. Older age was significantly associated with any abnormal finding, including arteriovenous malformations (P<0.0001); 50% of examinations included a therapeutic maneuver, most commonly argon plasma coagulation/cautery (43%). There were complications in 16 examinations (0.9%) including 2 perforations (0.1%), 6 cases with bleeding (0.3%) and 1 episode of pancreatitis (0.1%). CONCLUSIONS: DAE is most commonly performed to evaluate suspected small bowel bleeding and is commonly directed by video capsule findings. DAE is safe, has a high diagnostic yield, with 76% of examinations showing abnormal findings, and frequently features therapeutic maneuvers. Advancing age is associated with abnormal findings on DAE.

Use of Participatory Action Research in the Development of a Survey of Physiotherapy Services for People with Multiple Sclerosis in Canada.

Purpose: Currently, there is a paucity of research describing physiotherapy services for individuals with multiple sclerosis (MS) in Canada. Using qualitative methods, we aimed to develop a survey to examine physiotherapy practice patterns for people with MS receiving services in Canada. Method: We began by conducting a review of the current literature and combining participatory action research methods with the expertise of registered physiotherapists and individuals with MS. Semi-structured interviews were conducted with 10 participants to obtain their input into survey development. The interviews were then transcribed verbatim and analyzed thematically. Results: Five key themes emerged from the thematic analysis: (1) provide additional answer options, (2) reformat or clarify questions, (3) ensure that questions or options are appropriate, (4) ensure good readability and flow, and (5) determine the appropriate length of the survey. After a final revision, the survey consisted of 24 items in the following domains: demographics, MS programme and patient population, interdisciplinary care, and programme and service barriers. Conclusions: This survey is the first of its kind in Canada and is the first step toward improving the quality of health of people living with MS and the effectiveness of current physiotherapy practices for them.

Objectif : très peu d'études portent sur les services de physiothérapie pour les personnes atteintes de sclérose en plaques (SP) au Canada. À l'aide de méthodes qualitatives, la présente étude visait à préparer un sondage sur les modes d'exercice de la physiothérapie pour la SP au Canada. Méthodologie : analyse des publications à jour et combinaison de méthodes de recherche-action participatives avec les compétences de physiothérapeutes diplômés et de personnes atteintes de SP. Les chercheurs ont réalisé des entrevues semi-structurées avec dix participants pour obtenir leur avis sur l'élaboration du sondage. Les entrevues ont ensuite été transcrites textuellement, puis analysées par thèmes. Résultats : cinq thèmes principaux ont émergé de l'analyse thématique : 1) fournir d'autres possibilités de réponses, 2) reformuler ou clarifier les questions, 3) s'assurer que les questions ou les options sont appropriées, 4) s'assurer d'une bonne lisibilité et d'un bon enchaînement et 5) déterminer la bonne longueur du sondage. Après la dernière révision, le sondage se composait de 24 points dans les domaines suivants : démographie, programme pour la SP et population de patients, soins interdisciplinaires et obstacles aux programmes et aux services. Conclusion : le sondage est le premier du genre au Canada et représente la première étape vers l'amélioration de la qualité de vie des personnes ayant la SP ainsi que de l'efficacité des pratiques actuelles de physiothérapie auprès d'elles.

The effect of interval sprinting exercise on vascular function and aerobic fitness of post-menopausal women.

Menopausal transition accelerates an age-associated decrease in vascular function and a decline in aerobic fitness. The purpose of this study was to examine the effect of 8 weeks of interval sprinting cycle ergometer exercise on arterial stiffness, basal forearm blood flow, and aerobic fitness of post-menopausal women. Sixty overweight post-menopausal women were randomized into either exercise (Ex, n = 30) or control (C, n = 30) groups. Ex participants completed 24 interval sprinting exercise (ISE) sessions over 8 weeks. Each 20-minute ISE session comprised of alternating 8 seconds sprints and 12 seconds of light pedaling. Arterial stiffness assessed through ankle-brachial pulse wave velocity (baPWV) and augmentation index (AIx), basal forearm blood flow (FBF) assessed using venous occlusion, and aerobic fitness were assessed before and after the intervention. baPWV was significantly decreased in the Ex group by 7.2%, P = .03, whereas AIx demonstrated a 10% decrease, P = .002. No changes were found in basal FBF. Aerobic fitness was significantly increased, P = .002, in the Ex group (14%) with no change occurring in the control group.ISE training, despite minimal exercise commitment time (8 hours over 8 weeks), significantly lowered the arterial stiffness and increased the aerobic fitness of post-menopausal women. These results suggest that ISE positively influences the negative change in arterial stiffness and aerobic fitness that typically accompanies menopause.

Current Evidence for Acute Pain Management of Musculoskeletal Injuries and Postoperative Pain in Pediatric and Adolescent Athletes.

OBJECTIVE: Sports-related injuries in young athletes are increasingly prevalent with an estimated 2.6 million children and adolescents sustaining a sports-related injury annually. Acute sports-related injuries and surgical correction of sports-related injuries cause physical pain and psychological burdens on pediatric athletes and their families. This article aims to evaluate current acute pain management options in pediatric athletes and acute pain management strategies for postoperative pain after sports-related injuries. This article will also elucidate which areas of pain management for pediatric athletes are lacking evidence and help direct future clinical trials. DATA SOURCES: We conducted a literature search through PubMed and the Cochrane Central Register of Controlled Trials to provide an extensive review of initial and postoperative pain management strategies for pediatric sports-related musculoskeletal injuries. MAIN RESULTS: The current knowledge of acute pain management for initial sports-related injuries, postoperative pain management for orthopedic surgeries, as well as complementary and alternative medical therapies in pediatric sports-related injuries is presented. Studies evaluating conservative management, enteral and nonenteral medications, regional anesthesia, and complementary medical therapies are included. CONCLUSIONS: Adequate pain management is important for sports injuries in children and adolescents for emotional as well as physical healing, but a balance must be achieved to provide acceptable pain relief while minimizing opioid use and side effects from analgesic medications. More studies are needed to evaluate the efficacy of nonopioid analgesic medications and complementary therapies in pediatric patients with acute sports-related injuries.

On-demand manufacturing of clinical-quality biopharmaceuticals.

Conventional manufacturing of protein biopharmaceuticals in centralized, large-scale, single-product facilities is not well-suited to the agile production of drugs for small patient populations or individuals. Previous solutions for small-scale manufacturing are limited in both process reproducibility and product quality, owing to their complicated means of protein expression and purification. We describe an automated, benchtop, multiproduct manufacturing system, called Integrated Scalable Cyto-Technology (InSCyT), for the end-to-end production of hundreds to thousands of doses of clinical-quality protein biologics in about 3 d. Unlike previous systems, InSCyT includes fully integrated modules for sustained production, efficient purification without the use of affinity tags, and formulation to a final dosage form of recombinant biopharmaceuticals. We demonstrate that InSCyT can accelerate process development from sequence to purified drug in 12 weeks. We used integrated design to produce human growth hormone, interferon α-2b and granulocyte colony-stimulating factor with highly similar processes on this system and show that their purity and potency are comparable to those of marketed reference products.

Population well-being and electoral shifts.

Population wellbeing, an aggregate measure of positive mental, physical, and emotional health, has previously been used as a marker of community thriving. We examined whether several community measures of wellbeing, and their change since 2012, could be used to understand electoral changes that led to the outcome of the 2016 United States presidential election. We found that areas of the US which had the largest shifts away from the incumbent party had both lower wellbeing and greater drops in wellbeing when compared with areas that did not shift. In comparison, changes in income were not related to voting shifts. Well-being may be more useful in predicting and understanding electoral outcomes than some more conventional voting determinants.

EP4 Antagonism by E7046 diminishes Myeloid immunosuppression and synergizes with Treg-reducing IL-2-Diphtheria toxin fusion protein in restoring anti-tumor immunity.

Reprogramming of immunosuppressive tumor microenvironment (TME) by targeting alternatively activated tumor associated macrophages (M2TAM), myeloid-derived suppressor cells (MDSC), and regulatory T cells (Tregs), represents a promising strategy for developing novel cancer immunotherapy. Prostaglandin E2 (PGE2), an arachidonic acid pathway metabolite and mediator of chronic inflammation, has emerged as a powerful immunosuppressor in the TME through engagement with one or more of its 4 receptors (EP1-EP4). We have developed E7046, an orally bioavailable EP4-specific antagonist and show here that E7046 has specific and potent inhibitory activity on PGE2-mediated pro-tumor myeloid cell differentiation and activation. E7046 treatment reduced the growth or even rejected established tumors in vivo in a manner dependent on both myeloid and CD8+ T cells. Furthermore, co-administration of E7046 and E7777, an IL-2-diphtheria toxin fusion protein that preferentially kills Tregs, synergistically disrupted the myeloid and Treg immunosuppressive networks, resulting in effective and durable anti-tumor immune responses in mouse tumor models. In the TME, E7046 and E7777 markedly increased ratios of CD8+granzymeB+ cytotoxic T cells (CTLs)/live Tregs and of M1-like/M2TAM, and converted a chronic inflammation phenotype into acute inflammation, shown by substantial induction of STAT1/IRF-1 and IFNγ-controlled genes. Notably, E7046 also showed synergistic anti-tumor activity when combined with anti-CTLA-4 antibodies, which have been reported to diminish intratumoral Tregs. Our studies thus reveal a specific myeloid cell differentiation-modifying activity by EP4 blockade and a novel combination of E7046 and E7777 as a means to synergistically mitigate both myeloid and Treg-derived immunosuppression for cancer treatment in preclinical models.

Análisis de la estancia y la morbimortalidad en una unidad de neurocríticos tras la cirugía tumoral cerebral / Analysis of the stay and morbidity and mortality rates in a unit for neurocritical patients following surgery for a brain tumour

Objetivo. Evaluar la asociación entre las variables pre y postoperatorias con estancias superiores al día de ingreso y la morbimortalidad del paciente operado de tumor cerebral durante su estancia en una unidad de cuidados intensivos neurocríticos (UNC). Pacientes y métodos. El estudio retrospectivo incluyó una cohorte de 317 pacientes operados consecutivamente de tumor cerebral por diferentes neurocirujanos e ingresados en la UNC durante un período de tres años (2010-2012). Resultados. El 21,5% (n = 68) de los pacientes tuvo estancias superiores a un día (grupo L), y el 78,5% (n = 249), igual o menores a un día (grupo C). Se evaluó la asociación univariable de los factores de riesgo pre y perioperatorios con la estancia. No hubo diferencias significativas en los datos demográficos, el estado físico según la clasificación de la American Society of Anesthesiologists (ASA), las características anatomopatológicas ni el índice de gravedad tumoral radiológica entre los grupos L y C. Se necesitó intubación traqueal en el 42,6% (n = 29) de los pacientes del grupo L en algún momento del postoperatorio. El 19,1% (n = 13) de los pacientes del grupo L tuvo complicaciones sistémicas y regionales simultáneamente. Conclusiones. Existe una fracción importante de pacientes que tiene una estancia superior a un día en una UNC. Tanto la necesidad de intubación traqueal como la asistencia respiratoria y la aparición de complicaciones sistémicas y regionales pueden determinar estancias superiores a un día en una UNC (AU)

Aim. To evaluate the association between the pre- and post-operative variables with stays in hospital lasting more than one day and the morbidity and mortality rates of patients undergoing surgery for a brain tumour during their stay in a neurocritical intensive care unit (NCU). Patients and methods. The retrospective study, over a period of three years (2010-2012), involving a cohort of 317 patients who consecutively underwent surgical interventions due to brain tumours performed by different neurosurgeons and were hospitalised in the NCU. Results. A total of 21.5% (n = 68) of the patients were hospitalised for more than one day (group L), and 78.5% (n = 249) stayed for one day or less (group S). The univariable association of the pre- and post-operative risks with the length of stay was evaluated. There were no significant differences between groups L and S in terms of the demographic data, the physical status according to the classification of the American Society of Anesthesiologists (ASA), the pathological features or the radiological tumour severity index. Tracheal intubation was required in 42.6% (n = 29) of the patients in group L at some time during the post-operative period. Of the patients in group L, 19.1% (n = 13) had systemic and regional complications simultaneously. Conclusions. An important fraction of patients remain in an NCU for more than one day. The need for both tracheal intubation and respiratory assistance, together with the appearance of systemic and regional complications, can require stays in an NCU for more than one day (AU)

[Analysis of the stay and morbidity and mortality rates in a unit for neurocritical patients following surgery for a brain tumour]. / Análisis de la estancia y la morbimortalidad en una unidad de neurocríticos tras la cirugía tumoral cerebral.

AIM: To evaluate the association between the pre- and post-operative variables with stays in hospital lasting more than one day and the morbidity and mortality rates of patients undergoing surgery for a brain tumour during their stay in a neurocritical intensive care unit (NCU). PATIENTS AND METHODS: The retrospective study, over a period of three years (2010-2012), involving a cohort of 317 patients who consecutively underwent surgical interventions due to brain tumours performed by different neurosurgeons and were hospitalised in the NCU. RESULTS: A total of 21.5% (n = 68) of the patients were hospitalised for more than one day (group L), and 78.5% (n = 249) stayed for one day or less (group S). The univariable association of the pre- and post-operative risks with the length of stay was evaluated. There were no significant differences between groups L and S in terms of the demographic data, the physical status according to the classification of the American Society of Anesthesiologists (ASA), the pathological features or the radiological tumour severity index. Tracheal intubation was required in 42.6% (n = 29) of the patients in group L at some time during the post-operative period. Of the patients in group L, 19.1% (n = 13) had systemic and regional complications simultaneously. CONCLUSIONS: An important fraction of patients remain in an NCU for more than one day. The need for both tracheal intubation and respiratory assistance, together with the appearance of systemic and regional complications, can require stays in an NCU for more than one day.

TITLE: Analisis de la estancia y la morbimortalidad en una unidad de neurocriticos tras la cirugia tumoral cerebral.Objetivo. Evaluar la asociacion entre las variables pre y postoperatorias con estancias superiores al dia de ingreso y la morbimortalidad del paciente operado de tumor cerebral durante su estancia en una unidad de cuidados intensivos neurocriticos (UNC). Pacientes y metodos. El estudio retrospectivo incluyo una cohorte de 317 pacientes operados consecutivamente de tumor cerebral por diferentes neurocirujanos e ingresados en la UNC durante un periodo de tres años (2010-2012). Resultados. El 21,5% (n = 68) de los pacientes tuvo estancias superiores a un dia (grupo L), y el 78,5% (n = 249), igual o menores a un dia (grupo C). Se evaluo la asociacion univariable de los factores de riesgo pre y perioperatorios con la estancia. No hubo diferencias significativas en los datos demograficos, el estado fisico segun la clasificacion de la American Society of Anesthesiologists (ASA), las caracteristicas anatomopatologicas ni el indice de gravedad tumoral radiologica entre los grupos L y C. Se necesito intubacion traqueal en el 42,6% (n = 29) de los pacientes del grupo L en algun momento del postoperatorio. El 19,1% (n = 13) de los pacientes del grupo L tuvo complicaciones sistemicas y regionales simultaneamente. Conclusiones. Existe una fraccion importante de pacientes que tiene una estancia superior a un dia en una UNC. Tanto la necesidad de intubacion traqueal como la asistencia respiratoria y la aparicion de complicaciones sistemicas y regionales pueden determinar estancias superiores a un dia en una UNC.

SIRT1 knockdown promotes neural differentiation and attenuates the heat shock response.

Neurons have a limited capacity for heat shock protein (HSP) induction and are vulnerable to the pathogenic consequence of protein misfolding and aggregation as seen in age-related neurodegenerative diseases. Sirtuin 1 (SIRT1), an NAD(+) -dependent lysine deacetylase with important biological functions, has been shown to sustain the DNA-binding state of HSF1 for HSP induction. Here we show that differentiation and maturation of embryonic cortical neurons and N2a neuroprogenitor cells is associated with decreases in SIRT1 expression and heat shock-dependent induction of HSP70 protein. Tests of a pharmacological activator and an inhibitor of SIRT1 affirm the regulatory role of SIRT1 in HSP70 induction. Protein cross-linking studies show that nuclear SIRT1 and HSF1 form a co-migrating high molecular weight complex upon stress. The use of retroviral vectors to manipulate SIRT1 expression in N2a cells show that shRNA-mediated knock down of SIRT1 causes spontaneous neurite outgrowth coincident with reduced growth rate and decreased induction of hsp70-reporter gene, whereas SIRT1 over-expression blocks the induced neural differentiation of N2a cells. Our results suggest that decreased SIRT1 expression is conducive to neuronal differentiation and this decrease contributes to the attenuated induction of HSPs in neurons.

Novel small molecule inhibitors of TLR7 and TLR9: mechanism of action and efficacy in vivo.

The discovery that circulating nucleic acid-containing complexes in the serum of autoimmune lupus patients can stimulate B cells and plasmacytoid dendritic cells via Toll-like receptors 7 and 9 suggested that agents that block these receptors might be useful therapeutics. We identified two compounds, AT791 {3-[4-(6-(3-(dimethylamino)propoxy)benzo[d]oxazol-2-yl)phenoxy]-N,N-dimethylpropan-1-amine} and E6446 {6-[3-(pyrrolidin-1-yl)propoxy)-2-(4-(3-(pyrrolidin-1-yl)propoxy)phenyl]benzo[d]oxazole}, that inhibit Toll-like receptor (TLR)7 and 9 signaling in a variety of human and mouse cell types and inhibit DNA-TLR9 interaction in vitro. When administered to mice, these compounds suppress responses to challenge doses of cytidine-phosphate-guanidine (CpG)-containing DNA, which stimulates TLR9. When given chronically in spontaneous mouse lupus models, E6446 slowed development of circulating antinuclear antibodies and had a modest effect on anti-double-stranded DNA titers but showed no observable impact on proteinuria or mortality. We discovered that the ability of AT791 and E6446 to inhibit TLR7 and 9 signaling depends on two properties: weak interaction with nucleic acids and high accumulation in the intracellular acidic compartments where TLR7 and 9 reside. Binding of the compounds to DNA prevents DNA-TLR9 interaction in vitro and modulates signaling in vivo. Our data also confirm an earlier report that this same mechanism may explain inhibition of TLR7 and 9 signaling by hydroxychloroquine (Plaquenil; Sanofi-Aventis, Bridgewater, NJ), a drug commonly prescribed to treat lupus. Thus, very different structural classes of molecules can inhibit endosomal TLRs by essentially identical mechanisms of action, suggesting a general mechanism for targeting this group of TLRs.

Aberrant regulation and modification of heat shock factor 1 in senescent human diploid fibroblasts.

Induction of the heat shock response (HSR), determined by hsp70-luciferase reporter and HSP70 protein expression, is attenuated as a function of age of the IMR-90 human diploid fibroblasts. To better understand the underlying mechanism, we evaluated changes in the regulation and function of the HSF1 transcription factor. We show that the activation of HSF1 both in vivo and in vitro was decreased as a function of age, and this was attributable to a change in the regulation of HSF1 as the abundance of HSF1 protein and mRNA was unaffected. HSF1 was primarily cytosolic in young cells maintained at 37 degrees C, and heat shock promoted its quantitative nuclear translocation and trimerization. In old cells, some HSF1 was nuclear sequestered at 37 degrees C, and heat shock failed to promote the quantitative trimerization of HSF1. These changes in HSF1 could be reproduced by treating young cells with H2O2 to stunt them into premature senescence. Flow cytometry measurement of peroxide content showed higher levels in old cells and H2O2-induced premature senescent cells as compared to young cells. Experiments using isoelectric focusing and Western blot showed age-dependent changes in the mobility of HSF1 in a pattern consistent with its S-glutathiolation and S-nitrosylation; these changes could be mimicked by treating young cells with H2O2. Our results demonstrated dynamic age-dependent changes in the regulation but not the amount of HSF1. These changes are likely mediated by oxidative events that promote reversible and irreversible modification of HSF1 including S-glutathiolation and S-nitrosylation.

Negative regulation of monocyte adhesion to arterial elastic laminae by signal regulatory protein alpha and Src homology 2 domain-containing protein-tyrosine phosphatase-1.

Elastic laminae are extracellular matrix constituents that not only contribute to the stability and elasticity of arteries but also play a role in regulating arterial morphogenesis and pathogenesis. We demonstrate here that an important function of arterial elastic laminae is to prevent monocyte adhesion, which is mediated by the inhibitory receptor signal regulatory protein (SIRP) alpha and Src homology 2 domain-containing protein-tyrosine phosphatase (SHP)-1. In a matrix-based arterial reconstruction model in vivo, elastic laminae were resistant to leukocyte adhesion and transmigration compared with the collagen-dominant arterial adventitia. The density of leukocytes within the elastic lamina-dominant media was about 58-70-fold lower than that within the adventitia from 1 to 30 days. An in vitro assay confirmed the inhibitory effect of elastic laminae on monocyte adhesion. The exposure of monocytes to elastic laminae induced activation of SIRP alpha, which in turn activated SHP-1. Elastic lamina degradation peptides extracted from arterial specimens could also activate SIRP alpha and SHP-1. The knockdown of SIRP alpha and SHP-1 by specific small interfering RNA diminished the inhibitory effect of elastic laminae, resulting in a significant increase in monocyte adhesion. These observations suggest that SIRP alpha and SHP-1 potentially mediate the inhibitory effect of elastic laminae on monocyte adhesion.

Induction of morphological and biochemical apoptosis following prolonged mitotic blockage by halichondrin B macrocyclic ketone analog E7389.

E7389, a macrocyclic ketone analog of the marine natural product halichondrin B, currently is undergoing clinical trials for cancer. This fully synthetic agent exerts its highly potent in vitro and in vivo anticancer effects via tubulin-based antimitotic mechanisms, which are similar or identical to those of parental halichondrin B. In an attempt to understand the impressive potency of E7389 in animal models of human cancer, its ability to induce apoptosis following prolonged mitotic blockage was evaluated. Treatment of U937 human histiocytic lymphoma cells with E7389 led to time-dependent collection of cells in the G2-M phase of the cell cycle, beginning as early as 2 h and becoming maximal by 12 h. Increased numbers of hypodiploid events were seen beginning at 12 h, suggesting initiation of apoptosis after prolonged E7389-induced mitotic blockage. The identity of hypodiploid events as apoptotic cells under these conditions was confirmed by two additional morphologic criteria: green to orange/yellow shifts on acridine orange/ethidium bromide staining, and cell surface annexin V binding as assessed by flow cytometry. Several biochemical correlates of apoptosis also were seen following E7389 treatment, including phosphorylation of the antiapoptotic protein Bcl-2, cytochrome c release from mitochondria, proteolytic activation of caspase-3 and -9, and cleavage of the caspase-3 substrate poly(ADP-ribose) polymerase (PARP). In LNCaP human prostate cancer cells, treatment with E7389 also led to generation of hypodiploid cells, activation of caspase-3 and -9, and appearance of cleaved PARP, indicating that E7389 can activate cellular apoptosis pathways under anchorage-independent and -dependent cell culture conditions. These results show that prolonged mitotic blockage by E7389 can lead to apoptotic cell death of human cancer cells in vitro and can provide a mechanistic basis for the significant in vivo anticancer efficacy of E7389.

Anti-angiogenesis effects of borrelidin are mediated through distinct pathways: threonyl-tRNA synthetase and caspases are independently involved in suppression of proliferation and induction of apoptosis in endothelial cells.

Borrelidin, an antibiotic with anti-angiogenic activity, not only suppresses new capillary tube formation, but also collapses formed capillary tubes in a rat aorta culture model. Since it selectively inhibits threonyl-tRNA synthetase, we examined the effect of threonine on its anti-angiogenic activity. We found that a high concentration of threonine (1 mM) attenuated the ability of borrelidin to inhibit both capillary tube formation in the rat aorta culture model and human umbilical vein endothelial cells (HUVEC) proliferation, yet did not affect the ability of borrelidin to collapse formed capillary tubes or to induce apoptosis in HUVEC. Borrelidin activated caspase-3 and -8, and inhibitors of both caspase-3 and -8 suppressed borrelidin-induced apoptosis in HUVEC. Taken together, these data suggest that the anti-angiogenic effects of borrelidin are mediated through at least two mechanisms, i.e. one threonine-dependent and the other threonine-independent, and borrelidin induces apoptosis in endothelial cells via the caspase-8/-3 pathway.