Blocking ATP-releasing channels prevents high extracellular ATP levels and airway hyperreactivity in an asthmatic mouse model.

Allergic asthma is a chronic airway inflammatory response to different triggers like inhaled allergens. Excessive ATP in fluids from patients with asthma is considered an inflammatory signal and an important autocrine/paracrine modulator of airway physiology. Here, we investigated the deleterious effect of increased extracellular ATP (eATP) concentration on the mucociliary clearance (MCC) effectiveness and determined the role of ATP releasing channels during airway inflammation in an ovalbumin (OVA)-sensitized mouse model. Our allergic mouse model exhibited high levels of eATP measured in the tracheal fluid with a luciferin-luciferase assay and reduced MCC velocity determined by microspheres tracking in the trachea ex vivo. Addition of ATP had a dual effect on MCC, where lower ATP concentration (µM) increased microspheres velocity, whereas higher concentration (mM) transiently stopped microspheres movement. Also, an augmented ethidium bromide uptake by the allergic tracheal airway epithelium suggests an increase in ATP release channel functionality during inflammatory conditions. The use of carbenoxolone, a nonspecific inhibitor of connexin and pannexin1 channels reduced the eATP concentration in the allergic mouse tracheal fluid and dye uptake by the airway epithelium, providing evidence that these ATP release channels are facilitating the net flux of ATP to the lumen during airway inflammation. However, only the specific inhibition of pannexin1 with 10Panx peptide significantly reduced eATP in bronchoalveolar lavage and decreased airway hyperresponsiveness in OVA-allergic mouse model. These data provide evidence that blocking eATP may be a pharmacological alternative to be explored in rescue therapy during episodes of airflow restriction in patients with asthma.

Glycated Albumin Triggers an Inflammatory Response in the Human Airway Epithelium and Causes an Increase in Ciliary Beat Frequency.

The role of inflammation in airway epithelial cells and its regulation are important in several respiratory diseases. When disease is present, the barrier between the pulmonary circulation and the airway epithelium is damaged, allowing serum proteins to enter the airways. We identified that human glycated albumin (GA) is a molecule in human serum that triggers an inflammatory response in human airway epithelial cultures. We observed that single-donor human serum induced IL-8 secretion from primary human airway epithelial cells and from a cystic fibrosis airway cell line (CF1-16) in a dose-dependent manner. IL-8 secretion from airway epithelial cells was time dependent and rapidly increased in the first 4 h of incubation. Stimulation with GA promoted epithelial cells to secrete IL-8, and this increase was blocked by the anti-GA antibody. The IL-8 secretion induced by serum GA was 10-50-fold more potent than TNFα or LPS stimulation. GA also has a functional effect on airway epithelial cells in vitro, increasing ciliary beat frequency. Our results demonstrate that the serum molecule GA is pro-inflammatory and triggers host defense responses including increases in IL-8 secretion and ciliary beat frequency in the human airway epithelium. Although the binding site of GA has not yet been described, it is possible that GA could bind to the receptor for advanced glycated end products (RAGE), known to be expressed in the airway epithelium; however, further experiments are needed to identify the mechanism involved. We highlight a possible role for GA in airway inflammation.

A Report of the interdisciplinary education approach within the Universidad de La Frontera Dental Curriculum

ABSTRACT: Interprofessional education has become widely recognised as fundamental to educate new generations of dentists able to successfully join the dental public and private sectors and provide appropriate dental treatment to complex patients in a complex health system. This article presents the innovated curriculum of the dentistry course at Universidad de La Frontera describing its interprofessional education components. The description of IPE in the dental curriculum was done using Institutional documents. The university vocational training policy provides the framework in which IPE is implemented in undergraduate courses. Six out of eight dental curriculum components included IPE in their educational methodologies: 'Integrated units', 'Culture, society and mankind', 'Prevention', 'Professional Projections', 'Integrated clinics', and 'Profes sional Practice'. IPE is present thorough the dental course from first to final year and it takes different forms within each componen t. IPE is one of the methodologies used to achieve the learning outcomes stated in the curriculum. Passing through all modu- les of the dental curriculum leads to the development of generic and professional skills that allow students to obtain their professional title.

RESUMEN: La educación interdisciplinaria ha sido ampliamente reconocida como fundamental en la educación de nuevas generaciones de odontólogos capaces de unirse exitosamente a los sectores público y privado de salud y de otorgar tratamiento odontológico a pacientes con condiciones complejas en un sistema de salud complejo. Este artículo presenta el curriculum innovado de la carrera de odontología de la Universidad de La Frontera, describiendo sus componentes de educación interdisciplinaria (EID). Para la de EID en el plan de estudios se usaron una serie de documentos institucionales. La política de formación profesional da un marco en el cual EID es implementado en los planes de estudio de pregrado. Seis de ocho componentes del plan de estudios de la carrera de odontología incluyen EID en sus metodologías educativas: "Unidades de integración", "Cultura, hombre y sociedad", "Prevención", "Proyecciones profesionales", "Clínicas Integrales" y "Prácticas Profesionales. EID está presente a lo largo de toda la carrera de odontología del primer al último año y toma diferentes formas de acuerdo a cada componente. EID es una de las metodologías usadas para lograr los objetivos de aprendizaje establecidos en el plan de estudios. Cumplir con todas las asignaturas del plan lleva al desarrollo de habilidades genéricas y profesionales que permiten a los estudiantes obtener su título profesional.

Desde la Admisión Hasta el Ejercicio Profesional. Una Revisión de las Carreras de Odontología en Chile / From Admission to Professional Practice, A Review of Dental Programs in Chile

RESUMEN: Al año 1991 sólo existían tres carreras de odontología en Chile: Universidad de Chile (1911), Universidad de Concepción (1919) y la Universidad de Valparaíso (1955 como sede de la U. de Chile). Actualmente, existen 21 universidades que dictan un total de 32 carreras de odontología, titulando aproximadamente 1500 nuevos profesionales cada año, lo cual, aparte de disparar las cifras de profesionales en nuestro sistema, levanta la duda de cuantos estudiantes se encuentran en formación y cuál es el perfil de admisión que se establece en estas instituciones. El objetivo de este artículo es conocer las cifras de los procesos de admisión, matrícula financiamiento y acreditación universitarios y discutirlas en base a la cantidad de profesionales habilitados para trabajar en nuestro sistema para así reflejar la situación actual que vive la Odontología en Chile. Se realizó un estudio descriptivo de corte transversal realizado en base a la revisión de las bases estadísticas públicas. A pesar de sus altos costos asociados a la implementación de espacios y materiales, la carrera sigue siendo altamente postulada. Los nuevos ingresos rondan en alrededor de 2.20 0 para primer año, llegando aproximadamente a 14.300 estudiantes en todos los niveles para el 2019. El Estado de Chile debería abordar los problemas que se desprenden de estos indicadores, a través de una discusión seria y sistemática, incorporando diversos actores y basándose en información sobre la real necesidad de atención en salud de la población y la disponibilidad estructural del sistema de salu d público y privado para soportar determinado número de profesionales. La apertura de nuevas carreras de odontología se visualiza como un problema, mientras el país avanza a pasos lentos en una regulación que es requerida de manera urgente.

ABSTRACT: In 1991 there were only three dental programs in Chile: Universidad de Chile (1911), Universidad de Concepción (1919) and Universidad de Valparaíso (1955, as campus of U. de Chile). Currently, there are 21 universities that teach a total of 32 dental programs, graduating approximately 1500 new professionals each year, which, in addition to increasing the number of dentists in our system, raises the question as to how many students are in training, and what is the admission profile established by these institutions. The aim of this article is to know the number of admissions, registration, financing, and university accreditation process, based on the number of qualified professionals working in our system, in order to reflect the current situation of dentistry in Chile. A descriptive cross-sectional study was carried out based on the review of the public statistical resources. Despite the high cost of the program, associated with the implementation and materials, the program has many applicants. The number of enrolled students each year in the first semester is around 2200. Therefore, a high number of students are currently enrolled in all semesters, totalling approximately 14,300 in 2019. The Chilean government should address this problem based on information about the real need for healthcare of the population, within the framework of the public and private health system to support a certain number of professionals. The opening of new dental programs is viewed as a problem, while the country is advancing slowly in a regulation that is urgently required.

Lack of Kcnn4 improves mucociliary clearance in muco-obstructive lung disease.

Airway mucociliary clearance (MCC) is the main mechanism of lung defense keeping airways free of infection and mucus obstruction. Airway surface liquid volume, ciliary beating, and mucus are central for proper MCC and critically regulated by sodium absorption and anion secretion. Impaired MCC is a key feature of muco-obstructive diseases. The calcium-activated potassium channel KCa.3.1, encoded by Kcnn4, participates in ion secretion, and studies showed that its activation increases Na+ absorption in airway epithelia, suggesting that KCa3.1-induced hyperpolarization was sufficient to drive Na+ absorption. However, its role in airway epithelium is not fully understood. We aimed to elucidate the role of KCa3.1 in MCC using a genetically engineered mouse. KCa3.1 inhibition reduced Na+ absorption in mouse and human airway epithelium. Furthermore, the genetic deletion of Kcnn4 enhanced cilia beating frequency and MCC ex vivo and in vivo. Kcnn4 silencing in the Scnn1b-transgenic mouse (Scnn1btg/+), a model of muco-obstructive lung disease triggered by increased epithelial Na+ absorption, improved MCC, reduced Na+ absorption, and did not change the amount of mucus but did reduce mucus adhesion, neutrophil infiltration, and emphysema. Our data support that KCa3.1 inhibition attenuated muco-obstructive disease in the Scnn1btg/+ mice. K+ channel modulation may be a therapeutic strategy to treat muco-obstructive lung diseases.

Hallazgo radiográfico de cuerpo extraño en piso de seno maxilar de sujeto asintomático / Radiographic finding of a foreign body in the maxillary sinus floor of an asymptomatic subject

El seno maxilar (SM) es una cavidad par presente en el hueso maxilar, que puede presentar diversas alteraciones o lesiones patológicas factibles de ser pesquisadas mediante estudios imagenológicos. En este sentido, la opacidad, el ocupamiento y/o engrosamiento de las mucosas son los signos orientadores para el diagnóstico de patologías en estas estructuras, las que pueden ir desde sinusitis, hasta antrolitos, pasando por hipoplasias, pseudoquistes u osteomas. En el siguiente reporte presentamos el caso de una mujer de 75 años de edad en la cual mediante examinación radiográfica de rutina se observa un cuerpo extraño en el SM derecho en relación a diente 1.6. De acuerdo a esto, se solicitó una tomografía computarizada cone-beam (CBCT) para evaluar posibles implicancias con tratamiento dental rehabilitador. Al examen se observan dos estructuras de alta densidad (densidad metálica), redondeadas, una al lado de la otra, en contacto inmediato a la cortical sinusal sin comprometerla. También se observa una cortical sinusal conservada en todo su recorrido y discreto engrosamiento mucoso en relación a dichas estructuras. En consideración con los antecedentes de la paciente, se opta por un manejo conservador del hallazgo mediante controles periódicos. Se propone la hipótesis de la formación de un antrolito de origen exógeno, que se corresponde con el historial de tratamiento endodóntico en la zona. El diagnóstico diferencial de los hallazgos radiográficos encontrados es fundamental para establecer los lineamientos terapéuticos de nuestros pacientes, por ello es importante contar con personal clínico capacitado para la interpretación de las imágenes.

The maxillary sinus (SM) is an even cavity present in the maxillary bone, which may present a number of pathological alterations or lesions that can be investigated through imaging analysis. In this sense, opacity, location and / or thickening of the mucous membranes are guides for the diagnosis of pathologies in these structures. These can range from sinusitis, to antrolites, to hypoplasias, pseudocysts or osteomas. In this study the case of a 75-year-old woman is presented. During routine radiographic examination, a foreign body was observed in the right MS in relation to tooth 1.6. A cone-beam computed tomography (CBCT) was requested to evaluate possible rehabilitative dental treatment. Upon examination, two highdensity rounded structures (metal density) adjacent to each other were noted. The structures were in direct contact with the sinus cortex without compromising it. Furthermore in relation to the above structures, scant cortical sinus was observed throughout its course, as well as slight mucous thickening. Considering the patient's background, it was determined to use conservative treatment through periodic controls. Hypothesis of an exogenous mass is proposed, corresponding to the history of endodontic treatment in the area. In view of the differential diagnosis of radiographic findings found, it is essential to implement therapeutic guidelines for patients and have trained clinical staff available to interpret the images.

Recommendations for improving photo quality in close range photogrammetry, exemplified in hand bones of chimpanzees and gorillas / Recomendaciones para mejorar la calidad de las fotos en fotogrametría de corto alcance, ejemplificado en huesos de las manos de chimpancés y gorillas

SUMMARY: Photogrammetry is becoming increasingly popular in morphological research and teaching due to its portability, ability to reliably render 3D models, and quality-to-price relationship relative to some popular surface scanners. Compared to surface scanners, however, the learning process in photogrammetry can be very time consuming. Here we describe common mistakes of photo capture in close-range photogrammetry that greatly affect 3D output and tips to improve them. Problems were identified after the 3D model construction of 780 hand bones of chimpanzees and gorillas from museum collections. Their hands are composed of 27 bones which vary in length and complexity. We show how lighting, object position and orientation, camera angle, and background affect the 3D output. By taking these factors into account, time and error rates for beginners can be greatly reduced and 3D model quality can be considerably improved.

RESUMEN: La fotogrametría está siendo cada vez más popular en la investigación y enseñanza morfológica. Esto debido a su portabilidad, confiabilidad de los modelos 3D y buena relación calidadprecio. Comparada con los escáneres de superficie, sin embargo, el proceso de aprendizaje de la fotogrametría puede llevar mucho tiempo. Aquí se describen errores comunes en la toma de fotos para fotogrametería que afectan de manera importante la creación de los modelos 3D, así como consejos para superarlos. Los problemas descritos fueron identificados luego de la construcción de 780 modelos 3D de huesos de la mano de chimpancés y gorillas depositados en distintas colecciones de museos. Las manos de estas especies están compuestas por 27 huesos que varían en tamaño y complejidad. En este artículo mostramos como la luz, la posición y orientación del objeto, el ángulo de la cámara y el fondo de la imagen afectan el resultado en 3D. Considerando estos factores, personas que están aprendiendo esta técnica pueden reducir de manera importante el tiempo y la probabilidad de error, y mejorar considerablemente la calidad de los modelos 3D.

Comparison of morphological quantitative characteristics of the newly formed alveolar bone after the application of demineralized bone matrix and cortical bone allografts / Comparación de las características morfológicas cuantitativas del hueso alveolar recién formado después de la aplicación de matriz ósea desmineralizada y aloinjertos de hueso cortical

The allografts were used to obtain sufficient alveolar bone tissue for proper dental implant placement. The objective of the present study was to evaluate the morphological and quantitative characteristics (cellular and collagen densities) of the newly formed alveolar bone with the application of cortical bone (CB) and demineralized bone matrix (DBM) allografts. Six samples of alveolar bone tissue from 5 patients (50 ± 6.3 years) were obtained after 6 months of application of the allografts and immediately before the placement of the dental implants. The samples were fixed (buffered formaldehyde, pH7.2), decalcified (EDTA 10 %) and histologically processed (HE and Picro-Sirius) for histologic analysis. Morphological analysis revealed presence of osteocytes and trabeculae in neoformed bone tissue near the allografts and absence of inflammatory and allergic cells; the remnants of CB were located mainly in the periphery of the bone tissue and the remnants of DBM were more incorporated into the tissue. Osteogenitor cells were observed around the remaining material. The cell density was not modified in newly formed bone tissue with the application of both allografts as compared to mature bone tissue. The density of the type I and III collagens present in the osteoids interspersed with the remainder of the materials showed a tendency to increase in the samples treated with DBM. It was concluded that by the histological characteristics observed both grafts were biocompatible, however the bone treated with DBM presented better incorporation and a tendency of increase of the collagen content in the remnant region of the allografts.

Los aloinjertos son utilizados para obtener tejido óseo alveolar apropiado para la colocación correcta del implante dental. El objetivo de este trabajo fue evaluar las características morfológicas y cuantitativas (densidades celulares y de colágeno) del hueso alveolar recién formado con la aplicación de aloinjertos de hueso cortical (CB) y matriz desmineralizada de hueso (DBM). Seis muestras de tejido óseo alveolar fueron obtenidas de 5 pacientes (50 ± 6,3 años) después de 6 meses de aplicación de los aloinjertos e inmediatamente antes de la colocación de los implantes dentales. Las muestras fueron fijadas (formaldehído tamponado, pH 7,2), descalcificadas (EDTA al 10%) y procesadas histológicamente (HE y Picro-Sirius) para el análisis histológico. El análisis morfológico reveló la presencia de osteocitos y trabéculas en el tejido óseo neoformado cerca de los aloinjertos y la ausencia de células inflamatorias y alérgicas; los remanentes de CB se ubicaron principalmente en la periferia del tejido óseo y los remanentes de DBM se incorporaron más en el tejido. Se observaron células osteogenitoras alrededor del material restante. La densidad celular no se modificó en el tejido óseo recién formado con la aplicación de ambos aloinjertos en comparación con el tejido óseo maduro. La densidad de los colágenos de tipo I y III presentes en los osteoides intercalados con el resto de los materiales mostró una tendencia a aumentar en las muestras tratadas con DBM. Se concluyó que, debido a las características histológicas observadas, ambos injertos fueron biocompatibles, sin embargo, el hueso tratado con DBM presentó una mejor incorporación y una tendencia al aumento del contenido de colágeno en la región remanente de los aloinjertos.

Assessment of maxillary premolar region in relation to maxillary sinus floor and buccal bone plate: a cone beam computed tomography study / Evaluación de la región premolar maxilar en relación con el piso del seno maxilar y la tabla osea vestibular: estudio de tomografía computarizada de haz cónico

The maxillary sinus floor location and the buccal bone plate are factors to be considered in the long-term success with implant treatments mainly in the premolar region. the aim of this study was to establish morphometric characteristics of the buccal cortical bone (BCB) thickness of maxillary premolars and its relation to maxillary sinus floor through CBCT. In this study 350 first and second maxillary premolars were analyzed from 110 CBCT images and the buccal cortical bone (BCB) was measured in a coronal view at the major axis level of each premolar. In addition, in 200 first and second maxillary premolars CBCT images were measured the distance from premolar apex to maxillary sinus floor (MSF) in both frontal and sagittal plane. The type of relationship between the apex and MSF was classified according to Ok et al. (2014). The second premolar was observed with higher values of BCB (p<0.001). In first premolar, higher values were observed in the male sex (p>0.05). In second premolar, only significantly higher values were observed in the male sex in MA-MB-MC (p>0.05). Regarding to MSF and its relation to premolar roots, it was observed that 10 % of the sample was classified as type I, 19 % as type II, 55.5 % as type III and 15.5 % as type IV. The BCB of the upper premolar region is thicker in the apical region and decreases toward the coronal region. Almost 50 % of apex of second premolars are closely and risky related MSF (Type I and II).

La localización del piso del seno maxilar (PSM) y la tabla ósea vestibular (TOV) son factores a considerar en el éxito a largo plazo de los tratamientos de implante dental en la región premolar. El objetivo de este estudio fue establecer características morfométricas del grosor de la TOV en los premolares maxilares y su relación con el PSM a través de CBCT. Se analizaron 350 primeros y segundos premolares en 110 CBCT y la TOV fue medida en vista coronal en el eje axial mayor de cada premolar. Además en 200 primeros y segundos premolares maxilares se midió la distancia desde el ápice del diente hasta el PSM en el plano sagital y frontal. El tipo de relación entre el ápice y el PSM se clasificó según Ok et al. (2014). El Segundo premolar obtuvo los mayores valores de grosor de TOV (p<0.001). En primeros premolares se observaron valores altos en el género masculino (p>0.05). En segundos premolares solo se encontraron valores significativamente altos en el género masculino en MA-MB-MC (p>0.05). En la relación con el PSM y ápices de raíces de premolares, el 10% de la muestra se clasificó como tipo I, el 19% como tipo II, el 55,5% como tipo III y el 15.5% como tipo IV. En conclusión la TOV de la región premolares superior es más grueso en la zona apical, decreciendo hacia la zona coronal. Alrededor del 50% de los ápices de raíces de premolares maxilares están cercanas y en relación de riesgo con el PSM (tipo I y II).

Índices de simetría condilar y mandibular a través de radiografías panorámicas digitales en una muestra de pacientes chilenos / Condylar and mandibular symmetry indexes through digital panoramic radiographs in a sample of Chileans patients

La simetría mandibular puede alterarse por diversos motivos. Variados estudios buscan definir parámetros de "normalidad" y para ello se han descrito técnicas que utilizan la radiografía panorámica (RPN). El objetivo de este estudio fue determinar índices de simetría mandibular/condilar en una muestra de pacientes chilenos. Se realizó un estudio descriptivo de corte trasversal en RPN digitales obtenidas de la Clínica Odontológica de la Universidad de La Frontera. Se aplicaron criterios de inclusión: pacientes mayores de 18 años, dentición permanente completa hasta el primer molar; y de exclusión: signos radiográficos de alteraciones óseas y/ o cirugía mandibular, cóndilos no visibles, ortodoncia. Se utilizó el software ImageJ para dibujar rectas y medir la altura de la rama y cóndilo mandibular del lado derecho e izquierdo de acuerdo a la técnica propuesta por Kjellberg et al. (1994). Con estas medidas se calculó el índice de simetría entre altura condilar y rama (S1); y entre altura condilar y mandibular (S2). 235 RPN digitales fueron analizadas (95 hombres; 140 mujeres) con edad promedio de 24,98±7,32 años. El promedio de S1 fue de 93,74 % (±3,85) en hombres y 93,41 % (±5,77) en mujeres, mientras que para S2 fue de 90,50 % (±6,51) en hombres y 90,27 % (±7,80) en mujeres, sin diferencias significativas para ninguno de ambos índices (p=0,347 y p=0,233, respectivamente). En cuanto a la edad, tampoco se encontraron diferencias significativas en el promedio de los índices S1 y S2 para cada rango etario (p=0,594 y p=0,669, respectivamente). Mediante este estudio se concluye que hay un alto porcentaje de pacientes con simetría mandibular/condilar en ambos sexos y en diferentes rangos etarios, esto sin considerar el estado de salud o presencia de alteraciones morfológicas y/o funcionales.

Mandibular symmetry may be modified by various conditions. A number of studies have attempted to define parameters of "normality" and several techniques have been described using digital orthopantomography (OPG). The aim of this study was to determine mandibular/condylar symmetry indexes in a sample of Chilean patients. A cross sectional descriptive study in digital OPG obtained from the Dental Clinic of Universidad de La Frontera was performed. Inclusion criteria applied were: Patients older than 18 years, complete permanent dentition up to the first molar. Exclusion criteria were also applied: Radiographic signs of bone disease and/or mandibular surgery, not visible condyles, orthodontics. ImageJ software was used to draw lines and measure the height of the ramus and mandibular condyle on the right and left side with the technique postulated by Kjellberg (1994). With these measurements, the symmetry index between condylar height and ramus (S1) and between condylar and mandibular height (S2) were calculated. The study analyzed 235 digital OPG (95 men, 140 women) with an average age of 24.98 ± 7.32 years. The S1 average was 93.74 % (± 3.85) for men and 93.41 % (± 5.77) for women, and for S2 it was 90.50 % (± 6.51) for men and 90.27 % (± 7.80) for women, without significant differences for either index (p=0.347 and p=0.233, respectively). No significant differences were found in reference to age, for indexes S1 and S2 (p=0.594 and p=0.669, respectively). Therefore, it is concluded that there is presence of condylar/mandibular symmetry in both sexes and in age ranges, when measuring digital OPGs without considering health status or presence of any morphological or functional alteration.

Nitric oxide activation by progesterone suppresses ATP-induced ciliary activity in oviductal ciliated cells.

Ciliary beat frequency (CBF) regulates the oviductal transport of oocytes and embryos, which are important components of the reproductive process. Local release of ATP transiently increases CBF by increasing [Ca2+]i. Ovarian hormones also regulate ciliary activity and oviductal transport. Progesterone (P4) induces nitric oxide (NO) production and high P4 concentrations induce ciliary dysfunction. However, the mechanism by which P4 affects CBF has not been elucidated. To evaluate the role of P4 in NO production and its effect on ATP-induced increases in CBF, we measured CBF, NO concentrations and [Ca2+]i in cultures of oviductal ciliated cells treated with P4 or NO signalling-related molecules. ATP induced a [Ca2+]i peak, followed by an increase in NO concentrations that were temporally correlated with the decreased phase of the transiently increased CBF. Furthermore, P4 increased the expression of nitric oxide synthases (iNOS and nNOS) and reduced the ATP-induced increase in CBF via a mechanism that involves the NO signalling pathway. These results have improved our knowledge about intracellular messengers controlling CBF and showed that NO attenuates oviduct cell functions. Furthermore, we showed that P4 regulates neurotransmitter (ATP) actions on CBF via the NO pathway, which could explain pathologies where oviductal transport is altered and fertility decreased.

Bioluminiscencia: Herramienta de Medición y Análisis en Lavado de Manos Clínico Aplicado a la Odontología / Bioluminiscence Measurement Tool and Analysis in Clinical Hand Washing Technique Applied to Dentistry

RESUMEN: El estudio fue realizado en 40 estudiantes de ambos sexos, que cursaban 2° año el año 2017 de la carrera de Odontología, en estudiantes de la Universidad de La Frontera. El objetivo fue evaluar la efectividad de la técnica de Lavado de Manos Clínico (LMC) demostrando el grado de eficiencia, mediante Bioluminiscencia. La técnica que permite una medición cuantitativa rápida de los residuos orgánicos de superficie, por la cuantificación de cantidad de Adenosín Trifosfato (ATP). Empleándose un Luminómetro, 3M™ Clean-Trace™, se programó la medición de contaminación en dos puntos claves, región interdigital e hiponiquio, en ambas manos, por medio de la lectura de Unidades Relativas de Luz (URL). Se analizó la contaminación previa a la técnica de LMC y posterior al desarrollo de la técnica, en las regiones indicadas, por medio de Hisopado v/s aprobación/rechazo considerando como Aprobado <250 URL y Rechazado >251 URL., nivel de contaminación v/s sexo, y distribución de aprobación/rechazo tanto antes y después de realizada la técnica de LMC. El muestreo previo al LMC determinó en todas las localizaciones muestreadas existen rangos de aprobación/ rechazo entre 35-42,5 % / 57,5-65 %, encontrándose la mayor contaminación en ambas manos en hiponiquio. Posteriormente de realizada la técnica de LMC el rango de corte de aprobación/rechazo fue de 80-90 % / 10-20 %, el porcentaje más alto se pesquisó en mano izquierda hiponiquio. Para el análisis estadístico se utilizó el programa estadístico SPSS Statistics for Windows, análisis descriptivo, prueba t- test, prueba de Levene. Un valor de p < 0,05 para significancia. Obteniéndose diferencias significativas en análisis del valor la media, según zonas de muestreo y rangos de aprobación/rechazo en URL, que varían entre p=0,000 y p=0,015, que se presentó en mano izquierda zona interdigital posterior al LMC. Según la media concluimos que las regiones hisopadas tanto previas y posteriores a la ejecución del LMC en hiponiquio la piel está más contaminada, que es paradojal ya que la técnica incorpora un acento en esta localización, tiene un mejor acceso y contacto con jabón y agua.

SUMMARY: The study was conducted in 40 undergraduate students of both sexes, who were in the 2nd year of the 2017 year enrolled in the Undergraduate Dentistry Program of the Universidad de La Frontera. The objective was to evaluate the effectiveness of a Clinical Hand Washing technique (CML), demonstrating the degree of efficiency and implementing improvement behaviors of the technique using Bioluminescence. This technique allows a rapid quantitative measurement of organic residues on a surface by quantifying the amount of Adenosine Triphosphate (ATP). For that purpose, a Luminometer, 3M™ Clean-Trace™, was used to obtain the contamination level in two key points, Interdigital Region and Hyponychium, of both hands by quantification of Relative Light Units (URL). Hand contamination prior to CML and after the development of the technique was tested in the Interdigital and Hyponychium regions of both hands, through surface ATP swab considering as Approved <250 URL and Rejected > 251 URL. Also, the level of contamination v/s sex, and approval/rejection distribution before and after the CML were tested. The approval/rejection range for pre-CML sampling at all selected locations was 35-42.5 % / 57.5-65 %, fin ding the highest contamination rate in Hyponychium in both hands. After performing the CML we obtained 90 % of approval in all sites, with a 10 % rejection rate; the highest percentage was surveyed on left hand Hyponychium. Statistical analysis was performed using SPSS Statistics for Windows, a descriptive analysis, test t-test for independent samples, test of homogeneity of Levene variances. A p value <0.05 was established for statistical significance. There were significant differences in analysis of the mean value, according to sampling zones and URL approval / rejection ranges, ranging from p = 0.000 to p = 0.015 (Interdigital region of left hand, post-LA TÉCNICA DE LMC). The left hand presented greater contamination in both swab areas both before and after the execution of CML. The most contaminated region was the Hyponychium, which was paradoxical, because the technique emphasizes the hygiene of this location and has better access and contact with soap and water.

In vivo study on the slow release of glucose in vacuum fried matrices.

In vitro studies have shown that vacuum frying may be an effective process to reduce starch digestibility as it may limit gelatinization; this is significant as overconsumption of starchy foods contributes to obesity and type 2 diabetes. Although in vitro studies are an instrumental tool, in vivo studies allow observation of the overall effect on a living organism. The aim of this research was to assess how in vivo starch digestibility can be reduced when frying under vacuum (9.9 kPa), after feeding Sprague-Dawley rats, while also understanding its relationship to in vitro starch digestibility. Results showed that vacuum-fried dough has a lower degree of gelatinization (∼53.8%) and a maximal blood glucose level at 60 min (slower glycemic response) than atmospheric counterparts (∼98.3% degree of gelatinization and maximal blood glucose level at 30 min). Similarly, in vitro procedures exhibited less rapidly available glucose and higher unavailable glucose fractions in vacuum-fried dough.

An autocrine ATP release mechanism regulates basal ciliary activity in airway epithelium.

KEY POINTS: Extracellular ATP, in association with [Ca2+ ]i regulation, is required to maintain basal ciliary beat frequency. Increasing extracellular ATP levels increases ciliary beating in airway epithelial cells, maintaining a sustained response by inducing the release of additional ATP. Extracellular ATP levels in the millimolar range, previously associated with pathophysiological conditions of the airway epithelium, produce a transient arrest of ciliary activity. The regulation of ciliary beat frequency is dependent on ATP release by hemichannels (connexin/pannexin) and P2X receptor activation, the blockage of which may even stop ciliary movement. The force exerted by cilia, measured by atomic force microscopy, is reduced following extracellular ATP hydrolysis. This result complements the current understanding of the ciliary beating regulatory mechanism, with special relevance to inflammatory diseases of the airway epithelium that affect mucociliary clearance. ABSTRACT: Extracellular nucleotides, including ATP, are locally released by the airway epithelium and stimulate ciliary activity in a [Ca2+ ]i -dependent manner after mechanical stimulation of ciliated cells. However, it is unclear whether the ATP released is involved in regulating basal ciliary activity and mediating changes in ciliary activity in response to chemical stimulation. In the present study, we evaluated ciliary beat frequency (CBF) and ciliary beating forces in primary cultures from mouse tracheal epithelium, using videomicroscopy and atomic force microscopy (AFM), respectively. Extracellular ATP levels and [Ca2+ ]i were measured by luminometric and fluorimetric assays, respectively. Uptake of ethidium bromide was measured to evaluate hemichannel functionality. We show that hydrolysis of constitutive extracellular ATP levels with apyrase (50 U ml-1 ) reduced basal CBF by 45% and ciliary force by 67%. The apyrase effect on CBF was potentiated by carbenoxolone, a hemichannel inhibitor, and oxidized ATP, an antagonist used to block P2X7 receptors, which reduced basal CBF by 85%. Additionally, increasing extracellular ATP levels (0.1-100 µm) increased CBF, maintaining a sustained response that was suppressed in the presence of carbenoxolone. We also show that high levels of ATP (1 mm), associated with inflammatory conditions, lowered basal CBF by reducing [Ca2+ ]i and hemichannel functionality. In summary, we provide evidence indicating that airway epithelium ATP release is the molecular autocrine mechanism regulating basal ciliary activity and is also the mediator of the ciliary response to chemical stimulation.

TNFα Affects Ciliary Beat Response to Increased Viscosity in Human Pediatric Airway Epithelium.

In airway epithelium, mucociliary clearance (MCC) velocity depends on the ciliary beat frequency (CBF), and it is affected by mucus viscoelastic properties. Local inflammation induces secretion of cytokines (TNFα) that can alter mucus viscosity; however airway ciliated cells have an autoregulatory mechanism to prevent the collapse of CBF in response to increase in mucus viscosity, mechanism that is associated with an increment in intracellular Ca+2 level ([Ca2+]i). We studied the effect of TNFα on the autoregulatory mechanism that regulates CBF in response to increased viscosity using dextran solutions, in ciliated cells cultured from human pediatric epithelial adenoid tissue. Cultures were treated with TNFα, before and after the viscous load was changed. TNFα treatment produced a significantly larger decrease in CBF in cultures exposed to dextran. Furthermore, an increment in [Ca2+]i was observed, which was significantly larger after TNFα treatment. In conclusion, although TNFα has deleterious effects on ciliated cells in response to maintaining CBF after increasing viscous loading, it has a positive effect, since increasing [Ca2+]i may prevent the MCC collapse. These findings suggest that augmented levels of TNFα associated with an inflammatory response of the nasopharyngeal epithelium may have dual effects that contribute to maintaining the effectiveness of MCC in the upper airways.

New species of the earthworm genus Zapatadrilus (Clitellata, Acanthodrilidae) from northern Mexico.

Three new species of the earthworm genus Zapatadrilus are described from Tamaulipas, Mexico: Zapatadrilus aurelius sp. nov., Zapatadrilus huastecus sp. nov., and Zapatadrilus montezumensis sp. nov. Delimitation of the new species was supported by morphological and molecular (genetic distances) evidence. Diagnostic characters of the three new species include: tubulo-racemose prostates in segments 18 and 20, penial setae absent, typhlosole present and intestine beginning in 17/18. Z. montezumensis sp. nov. is separated by its metandric condition, Z. aurelius sp. nov. by the last hearts in 13 and Z. huastecus sp. nov. by the shape of the spermathecae and patterns of genital markings.

Low physiological levels of prostaglandins E2 and F2α improve human sperm functions.

Prostaglandins (PGs) have been reported to be present in the seminal fluid and cervical mucus, affecting different stages of sperm maturation from spermatogenesis to the acrosome reaction. This study assessed the effects of low physiological PGE2 and PGF2α concentrations on human sperm motility and on the ability of the spermatozoa to bind to the zona pellucida (ZP). Human spermatozoa were isolated from seminal samples with normal concentration and motility parameters and incubated with 1µM PGE2, 1µM PGF2α or control solution to determine sperm motility and the ability to bind to human ZP. The effects of both PGs on intracellular calcium levels were determined. Incubation for 2 or 18h with PGE2 or PGF2α resulted in a significant (P<0.05) increase in the percentage of spermatozoa with progressive motility. In contrast with PGF2α, PGE2 alone induced an increase in sperm intracellular calcium levels; however, the percentage of sperm bound to the human ZP was doubled for both PGs. These results indicate that incubation of human spermatozoa with low physiological levels of PGE2 or PGF2α increases sperm functions and could improve conditions for assisted reproduction protocols.

Regional-Scale Spatio-Temporal Analysis of Anastrepha ludens (Diptera: Tephritidae) Populations in the Citrus Region of Santa Engracia, Tamaulipas, Mexico.

Large citrus areas in Tamaulipas are affected by Anastrepha ludens (Loew) populations. Here we report the findings of a spatio-temporal analysis of A. ludens on an extended citrus area from 2008-2011 aimed at analyzing the probabilities of A. ludens infestation and developing an infestation risk classification for citrus production. A Geographic Information System combined with the indicator kriging geostatistics technique was used to assess A. ludens adult densities in the spring and fall. During the spring, our models predicted higher probabilities of infestation in the western region, close to the Sierra Madre Oriental, than in the east. Although a patchy distribution of probabilities was observed in the fall, there was a trend toward higher probabilities of infestation in the west than east. The final raster models summarized the probability maps using a three-tiered infestation risk classification (low-, medium-, and high risk). These models confirmed the greater infestation risk in the west in both seasons. These risk classification data support arguments for the use of the sterile insect technique and biological control in this extended citrus area and will have practical implications for the area-wide integrated pest management carried out by the National Program Against Fruit Flies in Tamaulipas, Mexico.

Independent anti-angiogenic capacities of coagulation factors X and Xa.

Knockout models have shown that the coagulation system has a role in vascular development and angiogenesis. Herein, we report for the first time that zymogen FX and its active form (FXa) possess anti-angiogenic properties. Both the recombinant FX and FXa inhibit angiogenesis in vitro using endothelial EA.hy926 and human umbilical cord vascular endothelial cells (HUVEC). This effect is dependent on the Gla domain of FX. We demonstrate that FX and FXa use different mechanisms: the use of Rivaroxaban (RX) a specific inhibitor of FXa attenuated its anti-angiogenic properties but did not modify the anti-angiogenic effect of FX. Furthermore, only the anti-angiogenic activity of FXa is PAR-1dependent. Using in vivo models, we show that FX and FXa are anti-angiogenic in the zebrafish intersegmental vasculature (ISV) formation and in the chick embryo chorioallantoic membrane (CAM) assays. Our results provide further evidence for the non-hemostatic functions of FX and FXa and demonstrate for the first time a biological role for the zymogen FX.

Calcium (Ca2+) waves data calibration and analysis using image processing techniques.

BACKGROUND: Calcium (Ca2+) propagates within tissues serving as an important information carrier. In particular, cilia beat frequency in oviduct cells is partially regulated by Ca2+ changes. Thus, measuring the calcium density and characterizing the traveling wave plays a key role in understanding biological phenomena. However, current methods to measure propagation velocities and other wave characteristics involve several manual or time-consuming procedures. This limits the amount of information that can be extracted, and the statistical quality of the analysis. RESULTS: Our work provides a framework based on image processing procedures that enables a fast, automatic and robust characterization of data from two-filter fluorescence Ca2+ experiments. We calculate the mean velocity of the wave-front, and use theoretical models to extract meaningful parameters like wave amplitude, decay rate and time of excitation. CONCLUSIONS: Measurements done by different operators showed a high degree of reproducibility. This framework is also extended to a single filter fluorescence experiments, allowing higher sampling rates, and thus an increased accuracy in velocity measurements.