Early Interaction of Alternaria infectoria Conidia with Macrophages.

Fungi of the genus Alternaria are ubiquitous indoor and outdoor airborne agents, and individuals are daily exposed to their spores. Although its importance in human infections and, particularly in respiratory allergies, there are no studies of how Alternaria spp. spores interact with host cells. Our aim was to study the early interaction of Alternaria infectoria spores with macrophages, the first line of immune defense. RAW 264.7 macrophages were infected with A. infectoria conidia, and the internalization and viability of conidia once inside the macrophages were quantified during the first 6 h of interaction. Live cell imaging was used to study the dynamics of this interaction. TNF-α production was quantified by relative gene expression, and the concentration of other cytokines (IL-1α, IL-1ß, IL-6, IL-4, IL-10, IL-17, GM-CSF and INF-γ) and a chemokine, MIP-1α, was quantified by ELISA. Conidia were rapidly internalized by macrophages, with approximately half internalized after 30 min of interaction. During the first 6 h of interaction, macrophages retained the ability to mitotically divide while containing internalized conidia. The classical macrophage-activated morphology was absent in macrophages infected with conidia, and TNF-α and other cytokines and chemokines failed to be produced. Thus, macrophages are able to efficiently phagocyte A. infectoria conidia, but, during the first 6 h, no effective antifungal response is triggered, therefore promoting the residence of these fungal conidia inside the macrophages.

Mechanistic study of the adjuvant effect of chitosan-aluminum nanoparticles.

The use of tailored particle-based adjuvants constitutes a promising way to enhance antigen-specific humoral and cellular immune responses. However, a thorough understanding of the mechanisms underlying their adjuvanticity is crucial to generate more effective vaccines. We studied the ability of chitosan-aluminum nanoparticles (CH-Al NPs), which combine the immunostimulatory effects of chitosan and aluminum salts, to promote dendritic cell activation, assess their impact on innate and adaptive immune responses, and compare the results to those reported for conventional chitosan particles (CH-Na NPs). All tested CH-NP formulations were capable of modulating cytokine secretion by dendritic cells. CH-Al NPs promoted NLRP3 inflammasome activation, enhancing the release of IL-1ß without significantly inhibiting Th1 and Th17 cell-polarizing cytokines, IL-12p70 or IL-23, and induced DC maturation, but did not promote pro-inflammatory cytokine production on their own. In vivo results showed that mice injected with CH-Al NPs generated a local inflammatory response comparable to that elicited by the vaccine adjuvant alum. Importantly, after subcutaneous immunization with CH-Al NPs combined with the hepatitis B surface antigen (HBsAg), mice developed antigen-specific IgG titers in serum, nasal and vaginal washes. Overall, our results established CH-Al NPs as a potential adjuvant to enhance both innate and adaptive immune responses.

Association of chitosan and aluminium as a new adjuvant strategy for improved vaccination.

The use of particulate adjuvants offers an interesting possibility to enhance and modulate the immune responses elicited by vaccines. Aluminium salts have been extensively used as vaccine adjuvants, but they lack the capacity to induce a strong cellular and mucosal immune response. Taking this into consideration, in this study we designed a new antigen delivery system combining aluminium salts with chitosan. Chitosan-aluminium nanoparticles (CH-Al NPs) exhibited a mean diameter of 280nm and a positive surface charge. The newly developed CH-Al NPs are more stable at physiological environment than classical CH NPs, showing no cytotoxic effects and revealing potential as a delivery system for a wide range of model antigens. In vivo studies showed that mice immunized with hepatitis B surface antigen (HBsAg)-containing CH NPs display high anti-HBsAg IgG titers in the serum, as well as the highest antigen-specific IgG on vaginal washes. Furthermore, in contrast to mice receiving antigen alone, mice immunized with the particulate adjuvant were able to elicit IgG2c antibody titers and exhibited higher antigen-specific IFN-γ levels in splenocytes. In conclusion, we established that CH-Al NPs, combining two immunostimulants to enhance both humoral and cellular immune responses, are a safe and promising system for antigen delivery. Our findings point towards their potential in future vaccination approaches.

Starch characterization in seven raw, boiled and roasted chestnuts (Castanea sativa Mill.) cultivars from Portugal.

Changes occurring in seven chestnut (Castanea sativa sp.) cultivars, caused by boiling and roasting, on starch content, cell and starch granules dimension were evaluated, and morphological changes were characterized by scanning electron microscopy. Three clear patterns of variation were detected after processing, namely: i) decrease of starch content with processing; ii) starch increase with the applied treatments; iii) increase of starch with boiling and decrease with roasting. Starch granules of raw chestnuts presented round, oval or elliptical form, external smooth surface and eccentric hilum, with rather ellipsoid-shaped growth rings. Processing resulted in modifications of the granules, with fusion of individual granules, and gelatinization taking place with the formation of elongated clusters. The present results indicate that boiling and roasting, besides changing the starch content of chestnut, causes important modifications in the starch granules, which can affect the sensory, rheological and chemical characteristics of chestnuts.

Intranasal Administration of Novel Chitosan Nanoparticle/DNA Complexes Induces Antibody Response to Hepatitis B Surface Antigen in Mice.

The generation of strong pathogen-specific immune responses at mucosal surfaces where hepatitis B virus (HBV) transmission can occur is still a major challenge. Therefore, new vaccines are urgently needed in order to overcome the limitations of existing parenteral ones. Recent studies show that this may be achieved by intranasal immunization. Chitosan has gained attention as a nonviral gene delivery system; however, its use in vivo is limited due to low transfection efficiency mostly related to strong interaction between the negatively charged DNA and the positively charged chitosan. We hypothesize that the adsorption of negatively charged human serum albumin (HSA) onto the surface of the chitosan particles would facilitate the intracellular release of DNA, enhancing transfection activity. Here, we demonstrate that a robust systemic immune response was induced after vaccination using HSA-loaded chitosan nanoparticle/DNA (HSA-CH NP/DNA) complexes. Furthermore, intranasal immunization with HSA-CH NP/DNA complexes induced HBV specific IgA in nasal and vaginal secretions; no systemic or mucosal responses were detected after immunization with DNA alone. Overall, our results show that chitosan-based DNA complexes elicited both humoral and mucosal immune response, making them an interesting and valuable gene delivery system for nasal vaccination against HBV.

Development of a novel adjuvanted nasal vaccine: C48/80 associated with chitosan nanoparticles as a path to enhance mucosal immunity.

In a time in which mucosal vaccines development has been delayed by the lack of safe and effective mucosal adjuvants, the combination of adjuvants has started to be explored as a strategy to obtain potent vaccine formulations. This study describes a novel adjuvant combination as an effective approach for a nasal vaccine - the association of the mast cell activator compound 48/80 with chitosan based nanoparticles. It was hypothesized that mucoadhesive nanoparticles would promote the cellular uptake and prolong the antigen residence time on nasal cavity. Simultaneously, mast cell activation would promote a local microenvironment favorable to the development of an immune response. To test this hypothesis, two different C48/80 loaded nanoparticles (NPs) were prepared: Chitosan-C48/80 NP (Chi-C48/80 NP) and Chitosan/Alginate-C48/80 NP (Chi/Alg-C48/80 NP). The potential as a vaccine adjuvant of the two delivery systems was evaluated and directly compared. Both formulations had a mean size near 500nm and a positive charge; however, Chi-C48/80 NP was a more effective adjuvant delivery system when compared with Chi/Alg-C48/80 NP or C48/80 alone. Chi-C48/80 NP activated mast cells at a greater extent, were better internalized by antigen presenting cells than Chi/Alg-C48/80 NP and successfully enhanced the nasal residence time of a model antigen. Superiority of Chi-C48/80 NP as adjuvant was also observed in vivo. Therefore, nasal immunization of mice with Bacillus anthracis protective antigen (PA) adsorbed on Chi-C48/80 NP elicited high levels of serum anti-PA neutralizing antibodies and a more balanced Th1/Th2 profile than C48/80 in solution or Chi/Alg-C48/80 NP. The incorporation of C48/80 within Chi NP also promoted a mucosal immunity greater than all the other adjuvanted groups tested, showing that the combination of a mast cell activator and chitosan NP could be a promising strategy for nasal immunization.

Multiple, unilateral lisch nodules in the absence of other manifestations of neurofibromatosis type 1.

Lisch nodules associated with Neurofibromatosis Type 1 (NF1) are usually multiple and bilateral in nature. Here, we report a 21-year-old healthy, Caucasian female who was diagnosed with multiple, unilateral Lisch nodules during routine eye examination. A thorough history and physical examination revealed no other signs of NF1. We diagnosed the rare occurrence of numerous, unilateral Lisch nodules in the absence of additional features of NF1 in our patient and provide a discussion concerning the differential diagnosis of Lisch nodules as well as the potential genetic explanation of this finding.

Sistema para el estudio de la circulación venosa periférica por Fotopletismografía. Validación clínica preliminar / System for the study of the peripheral venous circulation by Fotopletismografía: preliminary clinical validation

El trabajo presenta un sistema destinado para el estudio de la circulación venosa periférica por Fotopletismografía que se adicionará al equipo médico ANGIODIN PD 3000. El mismo permite calcular el Tiempo de Reflujo Venoso y a partir de éste realizar el diagnóstico de la insuficiencia valvular venosa en las extremidades inferiores. Se explican las características fundamentales del hardware empleado, que lo hacen idóneopara la prueba clínica de la bomba del músculo. Por último se realiza una comparación de la señal obtenida con la curva teórica. Se realizó una validación clínica preliminar obteniéndose parámetros estadísticos con un comportamiento positivo llegando a la conclusión de que el sistema es válido para la aplicación en el equipo médico ANGIODIN PD 3000(AU)

Autoinmunidad y función tiroidea en escolares de 2 regiones del Estado Mérida con deficiencia de yodo corregida

Objetivos: El presente estudio transversal fue planificado para evaluar la presencia de anticuerpo antitiroideo antiperoxidasa (ac.anti-TPO) y la función tiroidea, en dos áreas endémicas de bocio, con deficiencia de yodo corregida. Métodos: Se estudiaron 109 escolares, de ambos sexos, entre 6-12 años de edad, procedentes de Bailadores (BA=54) y Mucuchíes (MU=55). El diagnóstico de bocio se obtuvo por palpación clínica. La yoduria se determinó en 34 niños de BA y en 37 de MU, mientras que la concentración sérica de T4 libre (T4L), T3 libre (T3L), T3 total (T3T), TSH y ac.anti-TPO fueron determinadas en todos los escolares estudiados. Adicionalmente se realizó la cuantificación de T3T y ac.anti-TPO en uno de los progenitores de cada niño. Resultados: En BA, el 53,5% de los escolares presentaron bocio y en MU el 42,5%. La mediana de la excreción urinaria de yodo no fue estadísticamente diferente entre ambos grupos (BA=181 µg/L, MU=200 µg/L). La frecuencia global de detectabilidad de ac.anti-TPO en escolares fue del 45,5%, pero solo el 5% presentó seropositivad. Una concentración de ac. anti-TPO, entre 10 y 30 UI/mL, se observó en el 39% de los niños de BA y 40% en MU; 87% en adultos de BA y 54% en MU. En BA, concentraciones de ac.anti-TPO mayores de 30 UI/mL se observaron en el 10% de los niños y en el 17% de los adultos; en MU, no se observó ningún caso positivo para ac.anti-TPO en niños, mientras que en adultos la frecuencia de positividad fue del 8%. Las pruebas de función tiroidea de los escolares y de los progenitores de ambas regiones fueron similares y compatibles con estado de eufunción No hubo asociación entre la presencia de ac.anti-TPO y bocio. Conclusiones: Estos resultados indican que la corrección de la deficiencia de yodo en las áreas endémicas estudiadas se asocia con una alta frecuencia de detectabilidad de ac.anti-TPO, sin cambios en la función tiroidea.

Objective: This cross-sectional study was designed to evaluate the thyroid peroxidase autoantibody (TPO-Ab) concentrations and thyroid function in schoolchildren from two geographic regions were iodine deficiency was corrected. Methods: One hundred nine schoolchildren 6 to 12 years of age were studied: Bailadores (BA=54) and Mucuchíes (MU=55). Goiter size, urinary iodine, and serum freeT4, freeT3, total T3, TSH and TPO-Ab were measured. Total T3 and TPO-Ab were also determined in some parents. Results: Overall frequency of TPO-Ab detectability and positivity in schoolchildren were 45,5% and 5% respectively. TPO-Ab concentration ranging between 10-30 UI/mL was observed in 39% of children from BA and 40% from MU; 87% of parents from BA and 54% from MU. TPO-Ab concentration >30 UI/mL was observed in 10% of children and 17% of their parents. None of the children from MU had positive TPO-Ab, while 8% of their parents were positive. Median urinary iodine was similar in both regions (BA=181 µg/L, MU=200 µg/L). Thyroid function tests were normal in all cases. Goiter was found in 53,5% of children from BA and 42,5% from MU . No association between goiter and ac.TPO-Ab concentration was observed. Conclusions: These results suggest that in both endemic goiter regions, sufficient iodine prophylaxis by iodized salt is frequently associated with low detectable TPO-Ab levels and no changes in thyroid function.

Evaluation of the developmental toxicity of annatto in the rat.

Annatto, a dye extracted from Bixa orellana seeds, is used as a color additive in butter, cheese and in a variety of other foods as well as in drugs and cosmetics. Toxicological data on annatto and on its main carotenoid pigment bixin are still scarce. In this study we evaluated the developmental toxicity of annatto (28% of bixin). Annatto (0, 31.2, 62.5, 125, 250 and 500 mg/kg body weight/day) was given by gavage to Wistar rats on days 6-15 of pregnancy. Ceasarean sections were performed on day 21. Implantations, living and dead fetuses and resorptions were recorded. Fetuses were weighed and examined for externally-visible anomalies. One-third of fetuses from each litter was examined for visceral anomalies by a microsectioning technique. The remaining fetuses were cleared and stained with Alizarin Red S for skeleton evaluation. No adverse effect of annatto on the mothers was noted. No increase in embryolethality and no reduction of fetal body weight were observed among annatto-exposed rats. Annatto did not induce any increase in the incidence of externally-visible, visceral or skeletal anomalies in the exposed offspring. These findings suggest that annatto was neither maternally toxic nor embryotoxic in the rat. Therefore, the no-observed-adverse-effect level (NOAEL) for annatto-induced maternal and developmental toxicity was 500 mg/kg body weight/day or greater (or > or = 140 mg bixin/kg body weight/day) by the oral route.

Prevalencia del deficit nutricional de hierro y vitamina c en niños asistentes a círculos infantiles de la ciudad de Sancti Spíritus

Se obtuvo una muestra de 205 niños comprendidos entre 2,5 y 5,5 años de edad decimal procedentes de cinco círculos infantiles del municipio de Sancti Spíritus. Se les realizó hemoglobina, hematocrito, hierro sérico capacidad total de fijación de hierro(CTFH) y vitamina C sérica. Además se les calculó el porcentaje de saturación de la transferencia y la concentración media corpuscular hemoglobínica. También se efectuó un estudio dietético a una submuestra conformada por 97 niños. Se encontraron valores medios acordes a los criterios establecidos para todas las variables hematológicas estudiadas excepto la CTFH que resultó aumentada en ambos sexos. El 12,1 por ciento de las hembras y el 7,5 por ciento de los varones mostraron deficit de hierro, los cuales presentaron cifras significativamente menores de hemoglobina. Se constató un grupo considerable de niños con concentraciones de vitamina C en la categoria de alto riesgo.Por otra parte, la encuesta dietética arrojó que el consumo de hierro no cumple las recomendaciones por grupos etáreos, mientras la vitamina C sobrepasó lo estipulado.(AU)

Prevalencia del deficit nutricional de hierro y vitamina c en niños asistentes a círculos infantiles de la ciudad de Sancti Spíritus

Se obtuvo una muestra de 205 niños comprendidos entre 2,5 y 5,5 años de edad decimal procedentes de cinco círculos infantiles del municipio de Sancti Spíritus. Se les realizó hemoglobina, hematocrito, hierro sérico capacidad total de fijación de hierro(CTFH) yvitamina C sérica. Además se les calculó el porcentaje de saturación de la transferencia y la concentración media corpuscular hemoglobínica. También se efectuó un estudio dietético a una submuestra conformadapor 97 niños. Se encontraron valores medios acordes a los criterios establecidos para todas las variables hematológicas estudiadas excepto la CTFH que resultó aumentada en ambos sexos. El 12,1 por ciento de las hembras y el 7,5 por ciento de los varones mostraron deficit de hierro, los cuales presentaron cifras significativamente menores de hemoglobina. Se constató un grupo considerable de niños con concentraciones de vitamina C en la categoria de alto riesgo.Por otra parte, la encuesta dietética arrojó que el consumo de hierro no cumple las recomendaciones por grupos etáreos, mientras la vitamina C sobrepasó lo estipulado.(AU)

Enzyme activities in type I and II muscle fibres of human skeletal muscle in relation to age and torque development.

The quadriceps muscles from 20- 30- and 70-year-old clinically healthy men and women were studied regarding maximal isometric and isokinetic muscle torque in Newton metres (Nm), morphology and enzyme activity. Biopsy specimens were taken from the vastus lateralis muscle and freeze-dried, and individual fibres were dissected out and identified as type I or type II. The activities of citrate synthase (CS), 3-OHacyl-coA dehydrogenase (HAD), lactate dehydrogenase (LDH), myokinase (MK) and creatine phosphokinase (CPK) were determined in pools of type I and type II fibres. In both age groups a higher oxidative (CS, HAD, 1.3-1.5 x) and a lower glycolytic (LDH, 0.7 x) capacity was found in type I than in type II fibres. The myokinase activity was higher in type II (2 x) than in type I, whereas CPK activity was similar. The young men showed higher CS activity in both type I and type II fibres (1.5 x) and higher CPK activity in type I fibres (1.4 x) than the young women. There were only minor changes in oxidative or glycolytic capacities in relation to age. Myokinase was the only enzyme that decreased markedly with age in both pools of fibre types. Type II fibre area and mean fibre area correlated significantly to muscle torque in both sexes. In men, myokinase activity in type II fibres was significantly correlated to type II fibre area and to maximal muscle torque.

The quadriceps femoris muscle in 20-70-year-old subjects: relationship between knee extension torque, electrophysiological parameters, and muscle fiber characteristics.

Comparisons have been made between torque (isometric and isokinetic), electrophysiological (SFEMG, Macro EMG), and muscle fiber characteristics in the vastus lateralis muscle of both legs in healthy subjects aged between 20 and 70 years. Torque was greater in males and decreased with age in both sexes. Multifactorial analysis showed a positive correlation between torque, body surface area, and mean fiber area. These variables explained only about 30-40% of the torque changes. The electrophysiological parameters (Marco EMG amplitudes and fiber density) revealed evidence of reinnervation, indicating preceeding denervation and therefore loss of motor units. It was concluded that this fall out of motor units also contributes to the reduction in torque, when compensatory reinnervation begins to fall. Other factors, such as reduction in muscle fiber contractility, metabolic factors, and central factors, may also play a role in age-related reduction in torque.

Isometric and isokinetic knee extension and flexion torque in men and women aged 20-70.

An investigation was made of the influence of age and sex on peak muscle torque in knee extension and flexion during maximal isokinetic and isometric contraction. The study was performed on both legs of 139 clinically healthy men and 141 clinically healthy women aged 20, 30, 40, 50, 60 or 70 years. Maximum knee extension and flexion muscle torque (Newtonmetre, Nm) was measured isokinetically at various angles of velocity (12, 90 and 150 degrees/s) and isometrically under standardized conditions (Cybex II). No significant differences were found between the right and the left leg in the whole material. Muscle torque was higher in men than in women in all age groups (p less than 0.001). Both isokinetic and isometric torque decreased with age in both sexes. Isokinetic torque decreased significantly (p less than 0.05) between 20 and 30 years of age in men and between 40 and 50 years of age in women (at all velocities studied; p less than 0.05). A significant decrease (p less than 0.05) was found between the ages of 60 and 70 years in both sexes. Maximum isometric torque showed a significant decrease (p less than 0.05) between 60 and 70 years in men and women. There were no significant differences in isokinetic or isometric torque between moderately active and inactive men or women. Significant correlations were found between muscle torque and body weight, height and body surface area.

Estudio de los niveles de lípidos séricos y su relación con indicadores antropométricos en niños asistentes a círculos infantiles

En la ejecución de este trabajo nos propusimos conocer los niveles séricos de colesterol y triglicéridos y la relación existente con los indicadores antropométricos estudiados. Con tal propósito se obtuvo una muestra de 101 niños (52 varones y 49 hembras), provenientes de 4 circulos infantiles de la provincia a los que se les realizaron las determinaciones bioquímicas y las mediciones corporales que incluyeron: peso, estatura y pliegues cutáneos. Los valores medio de colesterol ascendieron a 169,5 mg/dl y 172 mg/dl para niñas y niños respectivamente. Se obtuvieron cifras de 63,9 mg/dl para los triglicéridos en las hembras y los varones alcanzaron 67 mg/dl para este parámetro. Se encontró correlación estadísticamente significativa en el sexo masculino y el colesterol y entre éste y los valores del pliegue tricipital situados en el P90 y por encima de éste de las normas nacionales.(AU)

Estudio de los niveles de lípidos séricos y su relación con indicadores antropométricos en niños asistentes a circulos infantiles

En la ejecución de este trabajo nos propusimos conocer los niveles séricos de colesterol y triglicéridos y la relación existente con los indicadores antropométricos estudiados. Con tal propósito se obtuvo una muestra de 101 niños (52 varones y 49 hembras), provenientes de 4 circulos infantiles de la provincia a los que se les realizaron las determinaciones bioquímicas y las mediciones corporales que incluyeron: peso, estatura y pliegues cutáneos. Los valores medio de colesterol ascendieron a 169,5 mg/dl y 172 mg/dl para niñas y niños respectivamente. Se obtuvieron cifras de 63,9 mg/dl para los triglicéridos en las hembras y los varones alcanzaron 67 mg/dl para este parámetro. Se encontró correlación estadisticamente significativa en el sexo masculino y el colesterol y entre éste y los valores del pliegue tricipital situados en el P90 y por encima de éste de las normas nacionales.(AU)

Histochemical and metabolic characteristics of human skeletal muscle in relation to age.

Muscle fibre composition, fibre areas and enzyme activities were determined in muscle biopsy samples (m. vastus lateralis) from both legs of 34 clinically healthy men and 31 clinically healthy women (aged 20-70 years). No significant difference was found in any of the three variables between the right and left leg. In the whole material the mean percentage of type I fibres was significantly higher in men, whereas that of type IIA fibres was significantly higher in women (men 58% I, 27% IIA, 13% IIB, women 51% I, 32% IIA, 15% IIB). The areas of all fibre types were significantly larger in men than in women. When the subjects were divided into age groups of 20, 30, 40, 50, 60 and 70 years, respectively, the fibre composition was found to be similar at different ages in both men and women, but changes in fibre areas were observed. The 70-year-old men and women showed significantly reduced areas of both type I and II fibres compared with the 60-year-olds. Only minor changes were seen in enzyme activities in relation to age. Men of the youngest age group had significantly higher levels of citrate synthase activity than those of the older age groups. Similar findings were seen for women except for the 60-year-olds, who had as high activity levels of citrate synthase as the 20-year-olds.

Muscle strength and balance in post-stroke patients.

Isokinetic torque of the knee and elbow during maximal extension and flexion and isometric strength of the handgrip, as well as balance and some circulatory variables, were tested in 37 patients one year after an acute stroke. Correlations with locomotion, household work and other activities of daily living (ADL) were tested. Male patients who had received special activation on the ward for up to 4 weeks after the stroke showed a significantly smaller difference in strength between the paretic and non-paretic knee at an angular velocity of 90 degrees per second than males who had received only routine activation on their ward. There was a high correlation between locomotion and the isokinetic torque of the paretic knee and also between locomotion and balance. The correlations between elbow strength and ADL functions were weaker. It was concluded that the evaluation of the isokinetic muscle torque of the knee and the balance tests are valid instruments for estimating functional capacity after a stroke.