Multitarget µ-Opioid Receptor Agonists─Neuropeptide FF Receptor Antagonists Induce Potent Antinociception with Reduced Adverse Side Effects.

The design of bifunctional compounds is a promising approach toward the development of strong analgesics with reduced side effects. We here report the optimization of the previously published lead peptide KGFF09, which contains opioid receptor agonist and neuropeptide FF receptor antagonist pharmacophores and is shown to induce potent antinociception and reduced side effects. We evaluated the novel hybrid peptides for their in vitro activity at MOP, NPFFR1, and NPFFR2 and selected four of them (DP08/14/32/50) for assessment of their acute antinociceptive activity in mice. We further selected DP32 and DP50 and observed that their antinociceptive activity is mostly peripherally mediated; they produced no respiratory depression, no hyperalgesia, significantly less tolerance, and strongly attenuated withdrawal syndrome, as compared to morphine and the recently FDA-approved TRV130. Overall, these data suggest that MOP agonist/NPFF receptor antagonist hybrids might represent an interesting strategy to develop novel analgesics with reduced side effects.

Aseguramiento y control de la calidad de los datos en un estudio de cohorte en Colombia

Introducción: La calidad de los datos facilita garantizar la fiabilidad de los estudios observacionales. Objetivo: Describir el aseguramiento y el control de calidad para mantener la fiabilidad y la validez del dato en un estudio de cohorte. Métodos: Presentar el manejo de datos implementado dentro de un seguimiento de enfermos renales crónicos cuya exposición fue un programa de protección renal comparado con el tratamiento convencional y su asociación con desenlaces clínicos. Se evaluó el cambio en la frecuencia de errores después de implementar el plan y la reproducibilidad del ingreso de registros a las bases de datos. Resultados: Se documentó una disminución progresiva en los errores cometidos en la captación de datos. El valor de Kappa entre los recolectores de la información para las variables clínicas más importantes fue 0,960 para la depuración de creatinina 150 mg/dL; 0,730 para la alteración del sedimento urinario; 0,956 para la asignación de estadio al ingreso. Los coeficientes de correlación intraclase para la identificación de las cifras de presión arterial sistólica fue 0,996; para la de presión arterial diastólica 0,993 y para los niveles de creatinina sérica al diagnóstico 0,995. Discusión: La calidad de los datos comienza con el reconocimiento de los retos y dificultades que implica su responsable captación, de ahí el aporte de la estandarización de los procesos y el personal que los lleve a cabo en forma idónea. Estudios evidencian que muchos procesos de mejora surgen en el desarrollo de la investigación sin protocolos preestablecidos. Conclusión: La reducción en la proporción y el tipo de error durante el proceso de captación de datos se debe a su identificación temprana y la corrección de instructivos, del instrumento de control de diligenciamiento y de la capacitación continua del personal. El análisis mostró una buena concordancia interevaluador.

Introduction: Data quality makes it easier to ensure that observational studies are reliable. Objective: To describe assurance and quality control to maintain data reliability and validity in a cohort study. Methodology: We present the data management strategies implemented in a study that followed patients of chronic kidney disease who were in a renal protection program and compared them with those undergoing conventional treatment to observe its association with clinical outcomes. We assessed the changes in error frequency after implementing the plan along with the reproducibility of the strategies for entering records into the databases. Results: We documented a progressive decrease of data collection errors. The Kappa values among data collectors for the most important variables were: 0.960 for creatinine clearance 150 mg/dl; 0.730 for urinary sediment alteration and 0.956 for stage allocation upon admission. The intraclass correlation coefficient for the identification of systolic blood pressure was 0.996; for diastolic blood pressure, the coefficient was 0.993 and for serum creatinine levels at diagnosis, the value was 0.995. Discussion: Data quality begins with the recognition of the challenges and difficulties involved in responsible data collection, hence the contribution of standardized processes and personnel to carry them out in a suitable manner. Studies show that many improvement processes arise in the development of research without pre-established protocols. Conclusion: The reduction in error ratio and type during the data collection process are the result of the early identification of erroneously entered or missing data, the correction of the guidelines for completing forms as well as of the instruments for detecting errors and continuous training of the staff. The analysis showed good inter-rater reliability.

Natural pentacyclic triterpenoid as allosteric modulators of human 5-lipoxygenase with potential anti-inflammatory activity.

This study explored new methods to inhibit human 5-lipoxygenase (5-hLOX) by analyzing natural terpenes that share structural similarities with acetoxyboswellic acid (AKBA). Enzymatic assays were used to evaluate the terpene's ability to inhibit the enzyme, potentially providing anti-inflammatory benefits. Our research focused on how certain types of triterpenes can inhibit 5-hLOX allosterically via a newly discovered allosteric site identified by enzyme crystallization. To determine whether natural boswellic acid analogs mimicked the allosteric known inhibitor AKBA, we combined 5-hLOX inhibition with in silico modeling. Our research has discovered that certain amino acids, specifically Arg 138, Arg 101, Arg 68, and Gln129, located in the allosteric 5-hLOX pocket, play a critical role in stabilizing glycyrrhetinic isomers. These amino acids form hydrogen bonds and hydrophobic interactions that contribute to the inhibitory potency of boswellic acid derivatives. We have found that α and ß glycyrrhetinic acid isomers, carbenoxolone, and to a minor extent, prednisolone, have a potent inhibitory effect against 5-hLOX with IC50 values of 8.64, 3.94, 52.98, and 291.20 µM, respectively. These values are in line with our calculated in silico allosteric site binding energy estimations. In contrast, other steroidal or non-steroidal anti-inflammatory agents exhibited inhibitory potencies larger than 500 µM. However, the specific pharmacodynamic mechanisms are currently unknown. We propose that AKBA analogs may lead to the future development of novel anti-inflammatory agents.Communicated by Ramaswamy H. Sarma.

Introduction of constrained Trp analogs in RW9 modulates structure and partition in membrane models.

Over the past decades, many cell-penetrating peptides (CPP) have been studied for their capacity to cross cellular membranes, mostly in order to improve cellular uptake of therapeutic agents. Even though hydrophobic and anionic CPPs have been described, many of them are polycationic, due to the presence of several arginine (Arg) residues. Noteworthy, however, the presence of aromatic amino acids such as tryptophan (Trp) within CPPs seems to play an important role to reach high membranotropic activity. RW9 (RRWWRRWRR) is a designed CPP derived from the polyarginine R9 presenting both features. In general, when interacting with membranes, CPPs adopt an optimal conformation for membrane interactions - an amphipathic helical secondary structure in the case of RW9. Herein, we assumed that the incorporation of a locally constrained amino acid in the peptide sequence could improve the membranotropic activity of RW9, by facilitating its structuration upon contact with a membrane, while leaving a certain plasticity. Therefore, two cyclized Trp derivatives (Tcc and Aia) were synthesized to be incorporated in RW9 as surrogates of Trp residues. Thus, a series of peptides containing these building blocks has been synthesized by varying the type, position, and number of modifications. The membranotropic activity of the RW9 analogs was studied by spectrofluorescence titration of the peptides in presence of liposomes (DMPG), allowing to calculate partition coefficients (Kp). Our results indicate that the partitioning of the modified peptides depends on the type, the number and the position of the modification, with the best sequence being [Aia4]RW9. Interestingly, both NMR analysis and molecular dynamic (MD) simulations indicate that this analog presents an extended conformation similar to the native RW9, but with a much-reduced structural flexibility. Finally, cell internalization properties were also confirmed by confocal microscopy.

Nanobody Loop Mimetics Enhance Son of Sevenless 1-Catalyzed Nucleotide Exchange on RAS.

RAS proteins control various intracellular signaling networks. Mutations at specific locations were shown to stabilize their active guanosine triphosphate (GTP)-bound state, which is associated with the development of multiple cancers. An attractive approach to modulate RAS signaling is through its regulatory guanine nucleotide exchange factor (GEF) son of sevenless 1 (SOS1). With the recent discovery of Nanobody14 (Nb14), which potently enhances SOS1-catalyzed nucleotide exchange on RAS, we explored the feasibility of developing peptide mimetics by structurally mimicking the complementarity-determining region 3 (CDR3). Guided by a biochemical GEF assay and X-ray co-crystal structures, successive rounds of optimization and gradual conformational rigidification led to CDR3 mimetics showing half of the maximal activation potential of Nb14 with an EC50 value of 29 µM. Altogether, this study demonstrated that peptides able to modulate a protein-protein interaction can be obtained by structural mimicry of a Nb paratope.

Diagnostic Performance of Airway Ultrasound for the Assessment of Difficult Laryngoscopy: A Systematic Review and Meta-Analysis.

OBJECTIVE: This study aimed to review and appraise the evidence regarding airway ultrasound assessment in predicting difficult laryngoscopy in adult patients. DESIGN: A systematic review of the literature was conducted according to the Cochrane collaboration guidelines and the recommendations for the systematic review and meta-analysis of diagnostic studies. Observational studies that evaluated the diagnostic performance of airway ultrasound for the prediction of difficult laryngoscopy were included for consideration. SETTING: Literature searches were performed in 4 databases (PubMed [Medline], Embase, Clinical Trials, and Google Scholar) to identify all observational studies using any ultrasound technique to assess difficult laryngoscopy. The search terms included "sonography," "ultrasound," "airway," "difficult airway," "difficult laryngoscopy," "Cormack," "risk factors," "ultrasound at the point of care," "difficult ventilation," "difficult intubation" and others, combined with sensitive filters. The search was done for studies performed in the last 20 years in English or Spanish. PARTICIPANTS: Adult patients older than 18 years old under general anesthesia for an elective procedure. Evident anatomic airway abnormalities, obstetric populations, those using an alternative imaging method besides ultrasound, and animal studies were excluded. INTERVENTIONS: Preoperative bedside ultrasound measuring distances and ratios from the skin to different reference points, such as the ratio of the hyomental distance in a neutral position (HMDN) and hyomental distance in extension (HMDR), HMDN, and the skin-to-epiglottis distance (SED), the preepiglottic area, and tongue thickness, among others. MEASUREMENTS AND MAIN RESULTS: A total of 24 studies evaluated the prediction of a difficult laryngoscopy using airway ultrasound. The diagnostic performance and the number of ultrasound parameters reported in the studies were variable. Meta-analysis was performed for 3 measurements consistently included in most studies. The SED and the HMDR ratio presented a sensitivity of 75% and 61%, respectively, and a specificity of 86% and 88%, respectively. The ratio of the preepiglottic distance to the epiglottic distance at the midpoint of the vocal cords (pre-E/E-VC) presented the best performance for predicting a difficult laryngoscopy (sensitivity: 82%, specificity: 83%, diagnostic odds ratio: 22.2). CONCLUSION: With the currently available evidence, the 3 commonly used point-of-care ultrasound measures used to identify difficult laryngoscopy, (SED, HMDR, and pre-E/E-VC), showed better sensitivity and similar specificity to clinical measures. Future studies and more data may change the authors' confidence in these conclusions, given the wide variability of measurements noted in studies.

Calidad de vida en pacientes con enfermedades tiroideas

Introducción: Las enfermedades del tiroides son relativamente frecuentes, constituyen un importante grupo dentro de las enfermedades crónicas no transmisibles. Objetivo: Determinar la calidad de vida en pacientes con enfermedades tiroideas. Métodos: Se realizó un estudio observacional, descriptivo y transversal. El universo estuvo constituido por 210 pacientes con diagnóstico de enfermedades tiroideas, que cumplieron con los criterios de inclusión y exclusión previstos en el estudio, previo consentimiento informado y la muestra no probabilística de 198 pacientes. Se utilizaron las variables: edad, sexo, enfermedad tiroidea diagnosticada (hipertiroidismo, hipotiroidismo, bocio difuso eutiroideo) y calidad de vida (facetas según dimensiones, según la clasificación de expertos y enfermedades tiroideas y la calidad de vida integral según enfermedades tiroideas). Se utilizó el cuestionario World Health Organization Quality of Life. Resultados: Predominó el hipotiroidismo como afección tiroidea más frecuente en las féminas entre 50 a 59 años. Las manifestaciones clínicas que puntuaron las medias más bajas fueron el dolor y malestar, seguido de indicaciones médicas y sentimientos negativos; las dimensiones físicas y psicológicas puntuaron con medias bajas, al igual que el ambiente, valores considerados como deficientes. Conclusiones: Predominó una calidad de vida integral media en el mayor por ciento de los pacientes, estas enfermedades deben ser identificadas a tiempo, para evitar otras complicaciones en diferentes sistemas del organismo que pudieran comprometer la vida del paciente.

Introduction: Thyroid diseases are relatively frequent; they constitute an important group within chronic non-communicable diseases. Objective: To determine the quality of life in patients with thyroid diseases. Methods: An observational, descriptive and cross-sectional study was carried out. The universe (210 patients) with a diagnosis of thyroid diseases, who met the inclusion and exclusion criteria provided for in the study, prior informed consent, and the non-probabilistic sample (198 patients). The variables used were: age, sex, diagnosed thyroid disease (hyperthyroidism, hypothyroidism, diffuse euthyroid goiter) and quality of life (facets according to dimensions, according to expert classification and thyroid diseases and comprehensive quality of life according to diseases thyroid). The World Health Organization Quality of Life questionnaire was used. Results: Hypothyroidism predominated as the most frequent thyroid condition in women between 50 and 59 years of age. The clinical manifestations that scored the lowest average were pain and discomfort, followed by medical indications and negative feelings; the physical and psychological dimensions scored with low averages, as well as the environment, values ​​considered deficient. Conclusions: An average comprehensive quality of life prevailed in the highest percentage of patients; these diseases must be identified in time, to avoid other complications in different body systems that could compromise the patient's life.

Unit Collective Medical Training in the 75th Ranger Regiment.

The 75th Ranger Regiment's success with eliminating preventable death on the battlefield is innate to the execution of a continuous operational readiness training cycle that integrates individual and unit collective medical training. This is a tactical solution to a tactical problem that is solved by the entire unit, not just by medics. When a casualty occurs, the unit must immediately respond as a team to extract, treat, and evacuate the casualty while simultaneously completing the tactical mission. All in the unit must maintain first responder medical skills and medics must be highly proficient. Leaders must be prepared to integrate casualty management into any phase of the mission. Leaders must understand that (1) the first casualty can be anyone; (2) the first responder to a casualty can be anyone; (3) medical personnel manage casualty care; and (4) leaders have ownership and responsibility for all aspects of the mission. Foundational to training is a command-directed casualty response system which serves as a forcing function to ensure proficiency and mastery of the basics. Four programs have been developed to train individual and collective tasks that sustain the Ranger casualty response system: (1) Ranger First Responder, (2) Advanced Ranger First Responder, (3) Ranger Medic Assessment and Validation, and (4) Casualty Response Training for Ranger Leaders. Unit collective medical training incorporates tactical leader actions to facilitate the principles of casualty care. Tactical leader actions are paramount to execute a casualty response battle drill efficiently and effectively. Successful execution of this battle drill relies on a command-directed casualty response system and mastery of the basics through rehearsals, repetition, and conditioning.

Anti-HCV and Zika activities of ribavirin C-nucleosides analogues.

Ribavirin is an unnatural nucleoside exhibiting broad spectrum of antiviral and antitumor activities, still very widely studied particularly in a repositioning approach. C-triazolyl nucleoside analogues of ribavirin have been synthesized, as well as prodrugs and glycosylated or peptide conjugates to allow a better activity by vectorization into the liver or by facilitating uptake into the cells. The antiviral properties of all synthesized compounds have been evaluated in vitro against two important human viral pathogens belonging to the Flaviviridae family: hepatitis C virus (HCV) and Zika virus (ZIKV). There are no therapeutic options for Zika virus, whereas those available for HCV can be still improved. Our results indicated that compound 2 carrying an N-hydroxy carboxamide function exhibits the most inhibitory activities against both viruses. This compound moderately inhibited the propagation of HCV with an IC50 value of 49.1 µM and Zika virus with an IC50 of 33.2 µM comparable to ribavirin in the Vero cell line. The results suggest that compound 2 and its new derivatives may be candidates for further development of new anti-HCV and anti-ZIKV antiviral drugs.

Identification and Characteristics of Fusion Peptides Derived From Enveloped Viruses.

Membrane fusion events allow enveloped viruses to enter and infect cells. The study of these processes has led to the identification of a number of proteins that mediate this process. These proteins are classified according to their structure, which vary according to the viral genealogy. To date, three classes of fusion proteins have been defined, but current evidence points to the existence of additional classes. Despite their structural differences, viral fusion processes follow a common mechanism through which they exert their actions. Additional studies of the viral fusion proteins have demonstrated the key role of specific proteinogenic subsequences within these proteins, termed fusion peptides. Such peptides are able to interact and insert into membranes for which they hold interest from a pharmacological or therapeutic viewpoint. Here, the different characteristics of fusion peptides derived from viral fusion proteins are described. These criteria are useful to identify new fusion peptides. Moreover, this review describes the requirements of synthetic fusion peptides derived from fusion proteins to induce fusion by themselves. Several sequences of the viral glycoproteins E1 and E2 of HCV were, for example, identified to be able to induce fusion, which are reviewed here.

Co-Treatment With Verapamil and Curcumin Attenuates the Behavioral Alterations Observed in Williams-Beuren Syndrome Mice by Regulation of MAPK Pathway and Microglia Overexpression.

Williams-Beuren syndrome (WBS) is a rare neurodevelopmental disorder characterized by a distinctive cognitive phenotype for which there are currently no effective treatments. We investigated the progression of behavioral deficits present in WBS complete deletion (CD) mice, after chronic treatment with curcumin, verapamil, and a combination of both. These compounds have been proven to have beneficial effects over different cognitive aspects of various murine models and, thus, may have neuroprotective effects in WBS. Treatment was administered orally dissolved in drinking water. A set of behavioral tests demonstrated the efficiency of combinatorial treatment. Some histological and molecular analyses were performed to analyze the effects of treatment and its underlying mechanism. CD mice showed an increased density of activated microglia in the motor cortex and CA1 hippocampal region, which was prevented by co-treatment. Behavioral improvement correlated with the molecular recovery of several affected pathways regarding MAPK signaling, in tight relation to the control of synaptic transmission, and inflammation. Therefore, the results show that co-treatment prevented behavioral deficits by recovering altered gene expression in the cortex of CD mice and reducing activated microglia. These findings unravel the mechanisms underlying the beneficial effects of this novel treatment on behavioral deficits observed in CD mice and suggest that the combination of curcumin and verapamil could be a potential candidate to treat the cognitive impairments in WBS patients.

Structure-Based Design of Melanocortin 4 Receptor Ligands Based on the SHU-9119-hMC4R Cocrystal Structure†.

The melanocortin receptors (MC1R-MC5R) belong to class A G-protein-coupled receptors (GPCRs) and are known to have receptor-specific roles in normal and diseased states. Selectivity for MC4R is of particular interest due to its involvement in various metabolic disorders, including obesity, feeding regulation, and sexual dysfunctions. To further improve the potency and selectivity of MC4R (ant)agonist peptide ligands, we designed and synthesized a series of cyclic peptides based on the recent crystal structure of MC4R in complex with the well-characterized antagonist SHU-9119 (Ac-Nle4-c[Asp5-His6-DNal(2')7-Arg8-Trp9-Lys10]-NH2). These analogues were pharmacologically characterized in vitro, giving key insights into exploiting binding site subpockets to deliver more selective ligands. More specifically, the side chains of the Nle4, DNal(2')7, and Trp9 residues in SHU-9119, as well as the amide linkage between the Asp5 and Lys10 side chains, were found to represent structural features engaging a hMC4R/hMC3R selectivity switch.

Case Series on 2g Tranexamic Acid Flush From the 75th Ranger Regiment Casualty Database.

Early tranexamic acid (TXA) administration for resuscitation of critically injured warfighters provides a mortality benefit. The 2019 Tactical Combat Casualty Care (TCCC) recommendations of a 1g drip over 10 minutes, followed by 1g drip over 8 hours, is intended to limit potential TXA side effects, including hypotension, seizures, and anaphylaxis. However, this slow and cumbersome TXA infusion protocol is difficult to execute in the tactical care environment. Additionally, the side effect cautions derive from studies of elderly or cardiothoracic surgery patients, not young healthy warfighters. Therefore, the 75th Ranger Regiment developed and implemented a 2g intravenous or intraosseous (IV/IO) TXA flush protocol. We report on the first six cases of this protocol in the history of the Regiment. After-action reports (AARs) revealed no incidences of post-TXA hypotension, seizures, or anaphylaxis. Combined, the results of this case series are encouraging and provide a foundation for larger studies to fully determine the safety of the novel 2g IV/IO TXA flush protocol toward preserving the lives of traumatically injured warfighters.

Optimized Opioid-Neurotensin Multitarget Peptides: From Design to Structure-Activity Relationship Studies.

Fusion of nonopioid pharmacophores, such as neurotensin, with opioid ligands represents an attractive approach for pain treatment. Herein, the µ-/δ-opioid agonist tetrapeptide H-Dmt-d-Arg-Aba-ß-Ala-NH2 (KGOP01) was fused to NT(8-13) analogues. Since the NTS1 receptor has been linked to adverse effects, selective MOR-NTS2 ligands are preferred. Modifications were introduced within the native NT sequence, particularly a ß3-homo amino acid in position 8 and Tyr11 substitutions. Combination of ß3hArg and Dmt led to peptide 7, a MOR agonist, showing the highest NTS2 affinity described to date (Ki = 3 pM) and good NTS1 affinity (Ki = 4 nM), providing a >1300-fold NTS2 selectivity. The (6-OH)Tic-containing analogue 9 also exhibited high NTS2 affinity (Ki = 1.7 nM), with low NTS1 affinity (Ki = 4.7 µM), resulting in an excellent NTS2 selectivity (>2700). In mice, hybrid 7 produced significant and prolonged antinociception (up to 8 h), as compared to the KGOP01 opioid parent compound.

Síndrome coronario agudo con elevación del segmento ST en el Centro de Diagnóstico Integral La Macandona / Acute coronary syndrome with ST elevation in La Macandona Comprehensive Diagnostic Center

RESUMEN Introducción: el síndrome coronario agudo con elevación del segmento ST, se debe a la oclusión completa de una de las arterias responsables del miocardio. Objetivo: determinar el comportamiento clínico epidemiológico del SCACEST en la sala de terapia intensiva del CDI La Macandona, del municipio Maracaibo, estado Zulia desde el año 2014 al 2017. Métodos: se realizó una investigación, observacional, longitudinal, descriptiva y retrospectiva, el universo de estudio fue los 82 portadores de la enfermedad. Se utilizaron las historias clínicas de cada paciente, las variables se agruparon en tablas según frecuencias absolutas y relativas. Resultados: predominó en el sexo masculino (73,17 %), en las edades de 60 a 69 años (55,84 %), como principal factor de riesgo la hipertensión arterial (80,48 %), la mayoría recibió tratamiento trombolítico con estreptoquinasa (88,23 %) y al final de la investigación el 96, 34 % fueron egresados vivos. Conclusiones: se deben diseñar protocolos de actuación en los centros de diagnóstico integral que garanticen una atención pronta y segura del SCACEST, para de e forma garantizar un mejor tratamiento y supervivencia en los pacientes portadores del mismo.

ABSTRACT Introduction: acute coronary syndrome with ST elevation is due to complete occlusion of one of the arteries responsible for the myocardium. Objective: to determine the clinical epidemiological behavior of acute coronary syndrome with ST elevation in the intensive care unit of La Macandona Comprehensive Diagnostic Center, Maracaibo municipality, Zulia state from 2014 to 2017. Methods: an observational, longitudinal, descriptive and retrospective study was conducted where the target group comprised 82 patients suffering from this pathology, the variables used: age, sex, smoking habit, hypercholesterolemia, diabetes mellitus, ischemic heart disease, whether they received thrombolytic treatment and hospital discharge status. To obtain the data the source used was the clinical histories of each patient; the variables were grouped in tables according to absolute and relative frequencies. Results: male sex 60 (73,17 %) predominated, 45 of them were from ages 60 to 69 (55,84 %), 66 suffered from hypertension (80,48 %) which was the main risk factor, most of them underwent thrombolytic treatment with streptokinase 75 (88,23 %) and 76 (96,34 %) were discharged alive from the hospital. Conclusions: protocols of actions must be designed in the Comprehensive Diagnostic Center that guarantee prompt and safe care, in order to guarantee better treatment and the survival of patients with acute coronary syndrome with ST elevation.

Neurotensin Analogues Containing Cyclic Surrogates of Tyrosine at Position 11 Improve NTS2 Selectivity Leading to Analgesia without Hypotension and Hypothermia.

Neurotensin (NT) exerts its analgesic effects through activation of the G protein-coupled receptors NTS1 and NTS2. This opioid-independent antinociception represents a potential alternative for pain management. While activation of NTS1 also induces a drop in blood pressure and body temperature, NTS2 appears to be an analgesic target free of these adverse effects. Here, we report modifications of NT at Tyr11 to increase selectivity toward NTS2, complemented by modifications at the N-terminus to impair proteolytic degradation of the biologically active NT(8-13) sequence. Replacement of Tyr11 by either 6-OH-Tic or 7-OH-Tic resulted in a significant loss of binding affinity to NTS1 and subsequent NTS2 selectivity. Incorporation of the unnatural amino acid ß3hLys at position 8 increased the half-life to over 24 h in plasma. Simultaneous integration of both ß3hLys8 and 6-OH-Tic11 into NT(8-13) produced a potent and NTS2-selective analogue with strong analgesic action after intrathecal delivery in the rat formalin-induced pain model with an ED50 of 1.4 nmol. Additionally, intravenous administration of this NT analogue did not produce persistent hypotension or hypothermia. These results demonstrate that NT analogues harboring unnatural amino acids at positions 8 and 11 can enhance crucial pharmacokinetic and pharmacodynamic features for NT(8-13) analogues, i.e., proteolytic stability, NTS2 selectivity, and improved analgesic/adverse effect ratio.

Studies of membranotropic and fusogenic activity of two putative HCV fusion peptides.

Over the past decades, membranotropic peptides such as positively charged cell-penetrating peptides (CPPs) or amphipathic antimicrobial peptides (AMPs) have received increasing interest in order to improve therapeutic agent cellular uptake. As far as we are concerned, we were interested in studying HCV fusion peptides as putative anchors. Two peptides, HCV6 and HCV7, were identified and conjugated to a fluorescent tag NBD and tested for their interaction with liposomes as model membranes. DSC and spectrofluorescence analyses demonstrate HCV7 propensity to insert or internalize in vesicles containing anionic lipids DMPG whereas no activity was observed with zwitterionic DMPC. This behavior could be explained by the peptide sequence containing a cationic arginine residue. On the contrary, HCV6 did not exhibit any membranotropic activity but was the only sequence able to induce liposomes' fusion or aggregation monitored by spectrofluorescence and DLS. This two peptides mild activity was related to their inefficient structuration in contact with membrane mimetics, which was demonstrated by CD and NMR experiments. Altogether, our data allowed us to identify two promising membrane-active peptides from E1 and E2 HCV viral proteins, one fusogenic (HCV6) and the other membranotropic (HCV7). The latter was also confirmed by fluorescence microscopy with CHO cells, indicating that HCV7 could cross the plasma membrane via an endocytosis process. Therefore, this study provides new evidences supporting the identification of HCV6 as the HCV fusion peptide as well as insights on a novel membranotropic peptide from the HCV-E2 viral protein.

Missed Connections at the Junction of Sociolinguistics and Speech Processing.

In recent years, significant momentum has built up in efforts to integrate the social with the cognitive in theoretical models of speech production/processing and phonological representation. While acknowledging these advances, we argue that what limits our ability to elaborate models of processing and representation in which social-indexical properties of speech are effectively integrated is that we remain some way from fully understanding how these properties are manifested within spoken interaction in the first place. We explore some of these limitations, drawing on data from a study of sociophonetic variability in a population of speakers of Australian English. We discuss issues relating to methods for capturing variability in the realization of vowels and consonants, and we highlight the pivotal role of speech style and the challenges that this raises for models of production and processing.

Caracterización clínico-epidemiológica de las infecciones respiratorias agudas en el menor de 5 años / Clinical and Epidemiological Characterization of Acute Respiratory Infections in Children under Five Years of Age

Las infecciones respiratorias agudas son un complejo sindromático de entidades clínicas, con diversidad epidemiológica y causal; causa más común de consulta médica pediátrica y de hospitalización. El objetivo fue caracterizar el comportamiento de las infecciones respiratorias agudas en pacientes pediátricos menores de 5 años ingresados en el Hospital Provincial Pediátrico Docente Pepe Portilla, Pinar del Río, durante el último semestre del año 2013. Se realizó una investigación observacional descriptiva y transversal. La muestra fue constituida por 88 pacientes pediátricos menores de 5 años. Se calcularon estadígrafos de tendencia central para expresar el comportamiento de variable cuantitativa. El sexo masculino reporta mayor frecuencia de infecciones respiratorias agudas. La lactancia materna no arrojó datos significativos, pero se observó una mayor incidencia de neumonías y bronconeumonía en pacientes que no la practicaban. La mayoría de los pacientes con enfermedades respiratorias agudas fueron niños entre 1 y 2 años. Los antecedentes patológicos personales y familiares de alergia y asma bronquial son importantes en la aparición de las enfermedades respiratorias agudas. Se hace necesario desarrollar acciones de promoción y prevención de salud para disminuir la incidencia de factores de riesgo y aparición de las infecciones respiratorias agudas en pacientes menores de 5 años, así como implementar algunas estrategias para reducir la morbilidad y mortalidad por estas enfermedades(AU)

Acute respiratory infections (ARIs) constitute a complex gamut of clinical features, with epidemiological and causal diversity, a most common cause for children medical consultation and for hospitalization. The objective was to characterize the behavior of ARIs in pediatric patients under 5 years of age admitted to Pepe Portilla Children Province Hospital of Pinar de Río Province, during the last semester of the year 2013. An observation, descriptive and cross-sectional research was carried out, with a sample constituted by 88 pediatric patients under five years of age. Statistic values of central tendency were calculated to express quantitative variable behavior.Results: the male sex reported a higher frequency of ARIs. Maternal breastfeeding did not show significant data, but a higher incidence was observed in pneumonia and bronchopneumonia. Most of the patients with ARIs were children aged between one and two years. Individual and family pathological history of allergy and bronchial asthma are important in the onset of AIRs. It is necessary to develop actions of health promotion and prevention to decrease the incidence of risk factor and the onset of ARIs in patient under five years of age, as well as to implement some strategies for reducing morbidity and mortality on this diseases(AU)

Daily egg consumption in hyperlipidemic adults--effects on endothelial function and cardiovascular risk.

BACKGROUND: Limiting consumption of eggs, which are high in cholesterol, is generally recommended to reduce risk of cardiovascular disease. However, recent evidence suggests that dietary cholesterol has limited influence on serum cholesterol or cardiac risk. OBJECTIVE: To assess the effects of egg consumption on endothelial function and serum lipids in hyperlipidemic adults. METHODS: Randomized, placebo-controlled crossover trial of 40 hyperlipidemic adults (24 women, 16 men; average age = 59.9 +/- 9.6 years; weight = 76.3 +/- 21.8 kilograms; total cholesterol = 244 +/- 24 mg/dL). In the acute phase, participants were randomly assigned to one of the two sequences of a single dose of three medium hardboiled eggs and a sausage/cheese breakfast sandwich. In the sustained phase, participants were then randomly assigned to one of the two sequences of two medium hardboiled eggs and 1/2 cup of egg substitute daily for six weeks. Each treatment assignment was separated by a four-week washout period. Outcome measures of interest were endothelial function measured as flow mediated dilatation (FMD) and lipid panel. RESULTS: Single dose egg consumption had no effects on endothelial function as compared to sausage/cheese (0.4 +/- 1.9 vs. 0.4 +/- 2.4%; p = 0.99). Daily consumption of egg substitute for 6 weeks significantly improved endothelial function as compared to egg (1.0 +/- 1.2% vs. -0.1 +/- 1.5%; p < 0.01) and lowered serum total cholesterol (-18 +/- 18 vs. -5 +/- 21 mg/dL; p < 0.01) and LDL (-14 +/- 20 vs. -2 +/- 19 mg/dL; p = 0.01). Study results (positive or negative) are expressed in terms of change relative to baseline. CONCLUSIONS: Egg consumption was found to be non-detrimental to endothelial function and serum lipids in hyperlipidemic adults, while egg substitute consumption was beneficial.