Correlation of frailty assessment metrics in one-year follow-up of aged care residents: a sub-study of a randomised controlled trial.

INTRODUCTION: Frailty is increasingly recognised as a dynamic syndrome, with multiple causes, dimensions and consequences. There is little understanding of how those frailty assessment metrics interact over time. The aim of this study was to describe the longitudinal correlation between five frailty metrics, namely multimorbidity, muscular strength, mood alterations, cognitive capacity, and functional capacity in a cohort study of aged care (nursing home) residents. METHODS: 248 aged care residents with Frailty Index at baseline of < 0.4 and no dementia were followed for 12 months. A multimorbidity score and an activity of daily living limitation score were created using individual items of the Frailty Index. Muscular strength was measured by grip strength. Cognitive capacity was measured using the Montreal Cognitive Assessment (MoCA) test. Mood alterations were measured using the anxiety/depression screening question from EQ-5D. We analysed the inter-individual correlation at baseline, association between baseline and future change, and within-individual correlation at baseline, 6 and 12 months. RESULTS: Population analysis shows that metrics were not associated at baseline. All of the studied metrics at baseline were associated with change in 12 months, with the exception of anxiety/depression scores. Pairwise within-individual correlation was strong between MoCA and grip strength (0.13, p = 0.02) and activity of daily living (- 0.48, p < 0.001), and between activities of daily living and multimorbidity index (0.28, p < 0.001). No within-individual correlation was found between anxiety depression score and other metrics. CONCLUSION: The results suggest an interdependence between comorbidities, physical capacity, cognition and activities of daily living in aged care residents. Comprehensive measurement of frailty-related metrics may provide improved understanding of frailty progression at later life stages.

Can different degrees of resistance training improve mood states in patients with fibromyalgia? A randomized controlled trial.

The purpose of this study was to look at the effects of low and high intensity resistance training (RT) on the mood states of fibromyalgia patients (FM). A total of 69 women participated in the study, with 31 healthy women serving as control group (HC), and 28 women diagnosed with FM being randomly assigned to one of two RT groups: low intensity or high intensity. Ten women diagnosed with FM formed a group of preferred intensity (PI). FM patients were subjected to 8 weeks of supervised RT with low, high, or PI doses. The exercise protocol was the same for both groups, with large muscle group exercises. Each intervention group performed a specific number of repetitions and rest periods based on the intensity. Training sessions took place twice a week. The HC received no type of intervention. The Brunel mood scale was used to assess mood states. When the mood profiles of patients with FM and healthy women were compared, patients with FM showed a worse mood profile. Low and high intensity RT for eight weeks did not improve the mood profile of FM patients. Anger showed a significant difference between LIRT and HIRT groups in the follow-up period (p=0.01); similarly significant differences between HIRT and HC were seen at baseline and at the 4 week evaluation in vigor (p=0.01 and p=0.001) and fatigue (p=0.01 and p=0.03). FM patients have a worse mood profile than healthy women, and eight weeks of low and high intensity RT did not result in significant improvements.

Food consumption on campus is associated with meal eating patterns among college students.

This is a cross-sectional analysis of data obtained in the baseline of the Longitudinal Study on the Lifestyle and Health of University Students (n 685) carried out in a public Brazilian university. Food intake was assessed using a 24-h dietary recall. Dietary patterns (DP) for breakfast, lunch and dinner were identified using principal component analysis. Generalised linear models were used to analyse the variables associated with each DP. Three DP were extracted for each meal: breakfast: 'White bread and butter/margarine', 'Coffee and tea' and 'Sausages, whole wheat bread and cheese'; lunch: 'Traditional', 'Western' and 'Vegetarian' and dinner: 'Beans, rice and processed juice', 'White bread and butter/margarine' and 'White meat, eggs and natural juice'. Students who had meals at the campus showed greater adherence to the 'White bread and butter/margarine' (exp (ßadj) = 1·15, 95 % CI 1·11, 1·19) and 'Coffee and tea' (exp (ßadj) = 1·06, 95 % CI 1·02, 1·10) breakfast patterns; 'Western' lunch pattern (exp (ßadj) = 1·04, 95 % CI 1·01, 1·08) and to the 'Beans, rice and processed juice' dinner pattern (exp (ßadj) = 1·10, 95 % CI 1·06, 1·14). Having meals at the campus was associated with lower adherence to the 'Sausages, whole wheat bread and cheese' breakfast pattern (exp (ßadj) = 0·93, 95 % CI 0·89, 0·97), 'Traditional' lunch pattern (exp (ßadj) = 0·96, 95 % CI 0·93, 0·99) and to the 'White bread and butter/margarine' (exp (ßadj) = 0·96, 95 % CI 0·93, 0·99) and 'White meat, eggs and natural juice' (exp (ßadj) = 0·96, 95 % CI 0·93, 0·99) dinner pattern. The food environment at campus may influence students' DP. Recognising meal eating patterns is important to support healthy eating promotion strategies on campus. Adjustments in the University Canteen menu could contribute to healthier eating choices among students.

Antifungal activity of extracts and purified saponins from the rhizomes of Chamaecostus cuspidatus against Candida and Trichophyton species.

AIM: To evaluate the antifungal activity of extracts of Chamaecostus cuspidatus against Candida and Trichophyton species. METHODS AND RESULTS: Crude ethanol extracts of leaves, stems and rhizomes were prepared and evaluated for antimicrobial activity. Only the rhizomes extract (RE) showed antifungal activity but had no inhibitory effect against bacteria (Staphylococcus aureus and Escherichia coli). The RE was then submitted to liquid-liquid partition with hexane (Hex), dichloromethane, chloroform, ethyl acetate and water. The Hex fraction (Hex Fr) from the RE was found to have the best antifungal effect. Three known saponins were isolated from the Hex Fr, of which two (dioscin and aferoside A) showed good antifungal activity. In addition, Hex Fr and the two bioactive compounds had no antibacterial effect, but exhibited fungicidal activity, caused significant changes in the morphology of the fungal cells and showed anti-Candida albicans biofilm activity. Finally, the bioactive plant products presented greater selectivity for fungal cells over normal human cells. CONCLUSIONS: The rhizomes of C. cuspidatus have bioactive saponins that function as effective antifungals against Candida and Trichophyton species, and have antibiofilm activity against C. albicans. SIGNIFICANCE AND IMPACT OF THE STUDY: Chamaecostus cuspidatus REs may have potential clinical application towards the management of superficial mycoses caused by Candida and Trichophyton species.

Long-term decline of the Amazon carbon sink.

Atmospheric carbon dioxide records indicate that the land surface has acted as a strong global carbon sink over recent decades, with a substantial fraction of this sink probably located in the tropics, particularly in the Amazon. Nevertheless, it is unclear how the terrestrial carbon sink will evolve as climate and atmospheric composition continue to change. Here we analyse the historical evolution of the biomass dynamics of the Amazon rainforest over three decades using a distributed network of 321 plots. While this analysis confirms that Amazon forests have acted as a long-term net biomass sink, we find a long-term decreasing trend of carbon accumulation. Rates of net increase in above-ground biomass declined by one-third during the past decade compared to the 1990s. This is a consequence of growth rate increases levelling off recently, while biomass mortality persistently increased throughout, leading to a shortening of carbon residence times. Potential drivers for the mortality increase include greater climate variability, and feedbacks of faster growth on mortality, resulting in shortened tree longevity. The observed decline of the Amazon sink diverges markedly from the recent increase in terrestrial carbon uptake at the global scale, and is contrary to expectations based on models.

Evaluating the menopausal transition with the STRAW + 10 in a Brazilian cohort of women with HIV, 2015-2016.

BACKGROUND: Menopausal transition is a physiological process encompassing hormonal and body changes that impact women's health and life quality. This period may be characterized by the Stages of Reproductive Aging Workshop (STRAW + 10) criteria using menstrual patterns. Use of the STRAW + 10 is uncertain in HIV infection. We aimed to characterize menopausal transition in women with HIV (WWH) using the STRAW + 10 criteria, hormonal measures and menopause symptoms. METHODS: We performed a cross-sectional study, nested to the HIV-Infected Women's Cohort, in Rio de Janeiro, Brazil. Eligible women included those aged 30 years or older, without clinical or surgical menopause, hormonal contraception, replacement therapy and ovarian disorders. We conducted face-to-face interviews and collected blood samples for follicle stimulating hormone (FSH) and estradiol measures. RESULTS: We enrolled 328 WWH (28.3% of women in the cohort). The distribution of age, hormonal levels and reported symptoms per each STRAW + 10 stage was consistent with the expected distribution in the menopausal transition. Age and FSH significantly increased and estradiol decreased from stage -2 (7 + days of menstrual delay) to stage +2 (8 + years of amenorrhea). CONCLUSIONS: The present results support use of the STRAW + 10 to characterize the menopausal transition of WWH with good clinical and immunological control.

First molecular evidence of frogs as a food source for sand flies (Diptera: Phlebotominae) in Brazilian caves.

Genus and species of phlebotomine sand flies have been recorded and described in caves in Brazil, but no study has provided the food source used by sand flies in these environments. Herein, we identified the blood source used by sand fly species in caves located at "Quadrilátero Ferrífero" (QF), Minas Gerais state. Specimens were manually collected near or on anurans inside ferruginous caves in Serra do Gandarela National Park and Serra do Rola Moça State Park. Males and females were placed in vials with 70% alcohol and stored at -10°C. Females engorged, after specific identification, had DNA extracted and followed for PCR amplification using specific primers. Sequencing was analyzed in the GenBank and Barcode of Life. A total of 198 specimens were collected (107 females and 91 males), all of them belonging to species Sciopemyia aff. microps (88.89%), Sciopemyia sordellii (10.61%), or Martinsmyia oliveirai (0.50%). When it comes to the females, 89 were S. aff. microps and 18 S. sordellii. Nineteen engorged females of S. aff. microps were analyzed and most of them (n=18) presented blood from Bokermannohyla martinsi and one contained blood from Scinax fuscovarius. The blood present in engorged females of S. sordellii (n=4) was from B. martinsi. Sciopemyia genus specimens are commonly found in collections carried out inside natural caves, but this was the first study to prove that females of this genus feed on cold-blooded animals in nature. HIGHLIGHTS: • Here we proved that sand flies feed in cold-blooded animals in in Brazilian caves. • Females of the Sciopemyia genus were for the first time found feeding in natural habitats. • Anurans of the family Hylidae were identified as source by molecular analyzes. • Insect bloodmeal identification can help assessing the fauna in several biomes. • This is the first record of S. aff. microps in caves of Brazil.

Effectiveness of Trichoderma strains isolated from the rhizosphere of citrus tree to control Alternaria alternata, Colletotrichum gloeosporioides and Penicillium digitatum A21 resistant to pyrimethanil in post-harvest oranges (Citrus sinensis L. (Osbeck)).

AIMS: Penicillium digitatum, Alternaria alternata and Colletotrichum gloeosporioides are pathogens responsible for large decays and production losses of citrus. They are commonly controlled by fungicides, whose excessive applications have led to the emergence of resistant P. digitatum strains. Alternative approaches are imperative for sustainable and environmental harmless citrus production, being biological control a promising strategy. The objective was to evaluate the potential of Trichoderma strains native from the rhizosphere of citrus trees to control these pathogens. METHODS AND RESULTS: Seven strains were isolated and identified as Trichoderma harzianum, T. guizhouense, T. atroviride and T. koningiopsis through morphological and molecular analyses. Five of them showed effective antagonist performance in vitro against the pathogens. The strain T. harzianum IC-30 was the best biological control agent in vivo, obtaining a reduction of rot percentage around 80% after 3 weeks of infection of oranges with P. digitatum A21 (resistant to pyrimethanil). This strain also showed the highest chitinase and glucanase activities. CONCLUSIONS: Trichoderma harzianum IC-30 is an optimal antagonist for the control of green mould spreading and other pathogens in post-harvest citrus fruits. SIGNIFICANCE AND IMPACT OF THE STUDY: The strain combined with supplementary practices could lead to sustainable management of citrus fungal diseases, dispensing with synthetic fungicides.

Preparation and evaluation of caffeine bioadhesive emulgels for cosmetic applications based on formulation design using QbD tools.

OBJECTIVE: The aim of the present study was to evaluate the influence of the incorporation of caffeine as a model active ingredient on the quality attributes of a bioadhesive emulgel formulation previously optimized by Quality by Design (QbD) tools. Emulgels are emerging topical drug delivery systems for cosmetic or pharmaceutical uses, which combine the advantages of both emulsions and gels. METHODS: In this work, the observed and predicted values for spreadability, phase separation by centrifugation and detachment forces performed by texturometer were compared with those of the control and active-containing emulgel formulations. In addition, rheological properties, release of caffeine and comparative in vitro/ex vivo bioadhesion properties were evaluated using human skin. RESULTS: The flow curves of emulgel formulations showed the typical pseudoplastic and no thixotropic flow with yield stress. The incorporation of active ingredient did not produce significant changes. All emulgels were uniformly spread and no significant differences in spreadability values between control and caffeine containing formulations and neither respect to those predicted values from experimental design optimization were found. Emulgel formulations showed appropriate detachment forces values and no significant differences between caffeine loading and control emulgel formulations were observed. CONCLUSION: Caffeine was successfully vehiculized in this optimized bioadhesive emulgel formulation, which showed high robustness regarding the process variability. There were no significant changes in the critical quality attributes after the incorporation of the active ingredient and a promising stability was observed for at least one year. Results suggested that the optimized emulgel is an interesting topical biodhesive delivery system for cosmetic applications, including agents for skin conditioning, not present in gels formulations.

OBJECTIF: la présente étude visait à évaluer l'influence de l'incorporation de la caféine comme une substance active modèle dans les attributs de qualité d'une préparation d'émulgel bioadhésif préalablement optimisée grâce à des outils QbD (Quality by Design, Qualité par la conception). Les émulgels sont des systèmes d'administration de médicaments topiques émergents destinés à des usages cosmétiques ou pharmaceutiques, qui associent les avantages des émulsions et des gels. MÉTHODES: dans cet étude, les valeurs observées et prédites en matière d'étalement, de séparation des phases par centrifugation et de forces de détachement par texturomètre ont été comparées à celles des préparations de contrôle et d'émulgel contenant des actifs. En outre, les propriétés rhéologiques, la libération de caféine et la comparaison des propriétés de bioadhérence in vitro / ex vivo ont été évaluées sur la peau humaine. RÉSULTATS: les courbes de débit des préparations d'émulgel révèlent un flux pseudoplastique typique et non thixotropique avec charge d'écoulement. L'incorporation de la substance active n'a pas produit de changements importants. Tous les émulgels ont été uniformément étalés et aucune différence significative dans les valeurs d'étalement entre les préparations de contrôle et celles contenant de la caféine n'a été observée. Aucune différence concernant les valeurs prédites dans l'optimisation de la conception expérimentale n'a été observée. Les préparations d'émulgel ont montré des valeurs de forces de détachement appropriées et aucune différence significative n'a été observée entre les préparations d'émulgel à base de caféine et les préparations de contrôle. CONCLUSION: la caféine a été administrée avec succès dans cette préparation optimisée d'émulgel bioadhésif, qui a affiché une grande résistance en ce qui concerne la variabilité du procédé. Il n'y a pas eu de changement significatif dans les attributs de qualité critiques après l'incorporation de la substance active et une stabilité prometteuse a été observée pendant au moins un an. Les résultats ont suggéré que l'émulgel optimisé est un système d'administration de bioadhérence topique intéressant pour les applications cosmétiques, y compris les agents de revitalisation de la peau qui ne se trouvent pas dans les préparations de gels.

A new acridone with antifungal properties against Candida spp. and dermatophytes, and antibiofilm activity against C. albicans.

AIM: The increase in the number of fungal infections worldwide, coupled with the limitations of current antifungal chemotherapy, demand the development of safe and effective new antifungals. Here, we presented the synthesis of a novel acridone (M14) and its antifungal properties against Candida and dermatophytes species. METHODS AND RESULTS: A series of 17 acridones was designed, synthesized and tested for its antifungal activity. The minimum inhibitory concentration (MIC) was determined by the broth microdilution method. Only the acridone M14 showed growth-inhibitory activity against reference strains and clinical isolates of Candida and dermatophytes, with MIC range of 7·81-31·25 µg ml-1 . Moreover, M14 exhibited fungicidal activity and prevented biofilm formation by C. albicans as well as reduced the viability of preformed biofilms, even at sub-MICs. The confocal laser scanning microscopy analysis revealed that C. albicans hyphal growth was completely inhibited in the presence of M14. Similarly, there was a severe inhibition on hyphal growth of Trichophyton rubrum. We also found that M14 has relatively low toxicity to human fibroblasts. CONCLUSIONS: The new acridone M14 has antifungal properties against Candida spp. and dermatophytes, and antibiofilm activity against C. albicans. In addition, M14 is relatively selective to fungal cells compared to human normal cells. SIGNIFICANCE AND IMPACT OF THE STUDY: Because of its in vitro antifungal activity, anti-Candida biofilm effect and moderate cytotoxicity towards normal human cell, M14 may serve as a valuable lead compound to develop a new antifungal agent.

High prevalence of non-alcoholic fatty liver disease and metabolic risk factors in Guatemala: A population-based study.

BACKGROUND: There are no data on the prevalence of non-alcoholic fatty liver disease (NAFLD) in general population samples in Guatemala or in other Central American countries. The prevalence and distribution of NAFLD and its associated risk factors were evaluated in a population-based sample of adults in Guatemala. METHODS: Cross-sectional study of 411 men and women 40 years of age or older residing in urban and rural areas of Guatemala. Metabolic outcomes included obesity, central obesity, hypercholesterolemia, diabetes, and metabolic syndrome (MetS). Liver disease outcomes included elevated liver enzymes, elevated Fatty Liver Index (FLI), and elevated FIB-4 score. RESULTS: The overall prevalence of obesity, central obesity, diabetes, and MetS were 30.9, 74.3, 21.6, and 64.2%, respectively. The fully-adjusted prevalence ratios (95% CI) for obesity, central obesity, diabetes, and MetS comparing women to men were 2.83 (1.86-4.30), 1.72 (1.46-2.02), 1.18 (1.03-1.34), and 1.87 (1.53-2.29), respectively. The overall prevalence of elevated liver enzymes (ALT or AST), elevated FLI, and elevated FIB-4 scores were 38.4, 60.1, and 4.1%, respectively. The fully-adjusted prevalence ratios (95% CI) for elevated liver enzymes (either ALT or AST) and elevated FLI score comparing women to men were 2.99 (1.84-4.86) and 1.47 (1.18-1.84), respectively. CONCLUSIONS: The prevalence of metabolic abnormalities and liver outcomes in this general population study was very high. The prevalence of metabolic and liver abnormalities was particularly high among women, an observation that could explain the atypical 1:1 male to female ratio of liver cancer in Guatemala.

Evaluation of the Induction of Cell-Mediated Immunity Against Candida albicans in a Model of Cutaneous Infection in Newborn 0-Day-Old Mice.

Candida albicans is a commensal fungus of the skin and mucous membranes in humans, but it is also responsible for mucocutaneous and systemic infections in immunocompromised patients like low birth weight neonates and premature newborns. The epicutaneous application of C. albicans is widely used to study the immune response against this pathogen in adult mice models. However, the immune response of newborns against infections caused by the genus Candida is poorly understood. In order to mimic premature human infection, we developed a model of C. albicans epicutaneous infection in newborn mice. We found that yeasts were able to colonize while the pseudohyphae invaded the epidermis. Recruitment of polymorphonuclear and mononuclear cells at the infection zone was observed. Fungal invasion, fungal burden and cellular infiltration displayed a time- and dose-dependent response. Interestingly, newborn mice were able to control C. albicans primary infection. Finally, we showed that the epicutaneous infection of C. albicans in newborn mice at birth results in the induction of cell-mediated immunity as evinced by delayed-type hypersensitivity assays.

Acute effect of strength training on mood of patients with fibromyalgia syndrome.

Fibromyalgia syndrome (FM) is a musculoskeletal disorder characterized by chronic pain and frequently associated changes in mood states. The aim of this study was to analyze the acute effect of strength training (ST) sessions on the mood states of patients with fibromyalgia. A total of 110 FM patients were eligible for this study. After the inclusion criteria, twenty-eight women with FM (mean age: 51.88±10.22 years) performed three sessions of ST. Each training session worked the main muscle groups and lasted 60 min. Three sets of 12 repetitions were performed with 1-min intervals between them. Outcome measures were assessed at baseline, after one session, and after three ST sessions. The Brunel Mood Scale (BRUMS) was used to assess mood states, and the Wilcoxon test was used to verify differences in mood after one and three ST sessions. The ST practice had positive effects on the patients' mood states after a single session. Reductions in anger, mental confusion, mood depression, fatigue, and tension were observed. The results of the 3rd ST session were similar. We concluded that a single ST session was sufficient to improve the mood states of patients with fibromyalgia.

Effect of photobiomodulation (670 nm) associated with vitamin A on the inflammatory phase of wound healing.

Wound healing is a complex biological process with specific phases. Photobiomodulation (PBM) decreases the inflammatory infiltrate, stimulating fibroblast proliferation and angiogenesis, and therefore, is indicated for wound healing. Vitamin A is used to reverse the inhibitory effects on wound healing and accelerate the healthy granulation tissue. The study aimed to evaluate the effect of topical vitamin A and PBM (GaAlAs) in inflammatory phase of cutaneous wounds. Forty Wistar male rats were separated into four groups: (1) control (CG); (2) laser group (LG) GaAlAs, 670 nm, 30 mW, energy per point of 0.9 J, radiating by 1 point in 30 s; (3) vitamin A group (VitAG); and (4) laser group plus vitamin A (LG + VitAG). Wounds were surgically made by a punch biopsy with 10 mm of diameter on the back of the animals and all treatments were started according to the experiment. The treatments were administered for four consecutive days and biopsy was performed on day 4. We performed both H&E and immunohistochemistry analysis. The results were compared between groups by one-way analysis of variance ANOVA test with post hoc Tukey (p < 0.05). Inflammatory infiltrate increased significantly in LG compared to CG and VitAG (p < 0.05). Regarding angiogenesis, VEGF expression was increased significantly in LG and LG + VitAG groups, p < 0.01. The results indicate that proposed treatments were effective on the healing process improved by LG and LG + VitAG. We show that laser plus vitamin A enhances healing by reducing the wound area and may have potential application for clinical management of cutaneous wounds.

Study of the Huntington's disease IT-15 gene in different ethnic groups in Ecuador.

This study aims to establish the current state of the IT-15 (HTT) gene in different Ecuadorian ethnic groups and patients by determining CAG triplet repeats, compared with the ethnicity of individuals. A total of 412 individuals were studied using nested polymerase chain reaction and Sanger sequencing: 75 individuals were indigenous (Kichwas), 211 mestizos, and 65 Afro-Ecuadorians. We included 31 patients who were clinically diagnosed with Huntington's disease (HD) and relatives of the affected patients (n = 30). Moreover, we correlated the presence of HD in Ecuadorian patients with 46 genetic ancestry-informative insertion-deletion polymorphic markers. We found that 77.20% had <28 CAG repetitions, 18.80% had mutable alleles, 2.27% had incomplete penetrance, and 1.70% reflected >39 repetitions. The average of CAG repetitions was 24 ± 3 for indigenous people; 28 ± 2 for mestizos; and 24 ± 3.2 repetitions for the Afro-Ecuadorians. The ancestral component showed that the main ancestry corresponded to Native Americans (0.873) and European ascendants (0.145), Africans were less represented in the evaluated population (0.018). There was a significant difference between the number of CAG repeats in mestizos and indigenous people (P < .01), suggesting that the Ecuadorian mestizo population has a risk factor for the gene mutation.

Sources and dynamics of fluorescent particles in hospitals.

Fluorescent particles can be markers of bioaerosols and are therefore relevant to nosocomial infections. To date, little research has focused on fluorescent particles in occupied indoor environments, particularly hospitals. In this study, we aimed to determine the spatial and temporal variation of fluorescent particles in two large hospitals in Brisbane, Australia (one for adults and one for children). We used an Ultraviolet Aerodynamic Particle Sizer (UVAPS) to identify fluorescent particle sources, as well as their contribution to total particle concentrations. We found that the average concentrations of both fluorescent and non-fluorescent particles were higher in the adults' hospital (0.06×106 and 1.20×106  particles/m3 , respectively) than in the children's hospital (0.03×106 and 0.33×106  particles/m3 , respectively) (P<.01). However, the proportion of fluorescent particles was higher in the children's hospital. Based on the concentration results and using activity diaries, we were able to identify sources of particle production within the two hospitals. We demonstrated that particles can be easily generated by a variety of everyday activities, which are potential sources of exposure to pathogens. Future studies to further investigate their role in nosocomial infection are warranted.

The role of wing geometric morphometrics in the identification of sandflies within the subgenus Lutzomyia.

The Lutzomyia subgenus (Diptera: Psychodidae) includes sibling species with morphologically indistinguishable females. The aims of this study were to analyse variations in the size and shape of wings of species within the Lutzomyia subgenus and to assess whether these analyses might be useful in their identification. Wings (n = 733) of 18 species deposited in Brazilian collections were analysed by geometric morphometrics, using other genera and subgenera as outgroups. Shape variation was summarized in multivariate analyses and differences in wing size among species were tested by analysis of variance. The results showed significant variation in the sizes and shapes of wings of different Lutzomyia species. Two clusters within the Lutzomyia subgenus were distinguished in analyses of both males and females. In Cluster 1 (Lutzomyia ischnacantha, Lutzomyia cavernicola, Lutzomyia almerioi, Lutzomyia forattinii, Lutzomyia renei and Lutzomyia battistinii), scores for correct reclassification were high (females, kappa = 0.91; males, kappa = 0.90), whereas in Cluster 2 (Lutzomyia alencari, Lutzomyia ischyracantha, Lutzomyia cruzi, Lutzomyia longipalpis, Lutzomyia gaminarai and Lutzomyia lichyi), scores for correct reclassification were low (females, kappa = 0.42; males, kappa = 0.48). Wing geometry was useful in the identification of some species of the Lutzomyia subgenus, but did not allow the identification of sibling species such as L. longipalpis and L. cruzi.

Responsiveness of pituitary to galanin throughout the reproductive cycle of male European sea bass (Dicentrarchus labrax).

The neuropeptide galanin (Gal) is a putative factor regulating puberty onset and reproduction through its actions on the pituitary. The present study investigated the pituitary responsiveness to galanin and the patterns of galanin receptors (Galrs) expression throughout the reproductive cycle of two years old male European sea bass (Dicentrarchus labrax), an important aquaculture species. Quantitative analysis of pituitary and hypothalamus transcript expression of four galr subtypes revealed differential regulation according to the testicular developmental stage, with an overall decrease in expression from the immature stage to the mid-recrudescence stage. Incubation of pituitary cells with mammalian 1-29Gal peptide induced significant changes in cAMP concentration, with sensitivities that varied according to the testicular development stages. Furthermore 1-29Gal was able to stimulate both follicle stimulating hormone (Fsh) and luteinizing hormone (Lh) release from pituitary cell suspensions. The magnitude of the effects and effective concentrations varied according to reproductive stage, with generalized induction of Fsh and Lh release in animals sampled in January (full spermiation). The differential expression of galrs in pituitary and hypothalamus across the reproductive season, together with the differential effects of Gal on gonadotropins release in vitro strongly suggests the involvement of the galaninergic system in the regulation the hypothalamus-pituitary-gonad axis of male sea bass. This is to our knowledge the first clear evidence for the involvement of galanin in the regulation of reproduction in non-mammalian vertebrates.

Analysis of tryptase-positive mast cells and immunoexpression of MMP-9 and MMP-13 in periapical lesions.

AIM: To evaluate the immunoexpression of tryptase, MMP-9 and MMP-13 in periapical lesions, correlating them with the type of lesion, intensity of the inflammatory infiltrate and thickness of the epithelial lining. METHODOLOGY: Twenty periapical granulomas (PGs), twenty radicular cysts (RCs) and twenty residual radicular cysts (RRCs) were submitted to immunohistochemical analysis using antitryptase, anti-MMP-9 and anti-MMP-13 antibodies. Immunoexpression of MMP-9 and MMP-13 was quantitatively evaluated both in the connective tissue of all lesions and in the epithelial lining of RCs and RRCs. Tryptase-positive mast cells were counted only in the connective tissue. The data were analysed using Kruskal-Wallis and Fisher's exact tests, as well as Spearman's correlation test (P < 0.05). RESULTS: In comparison with RCs and RRCs, PGs exhibited higher immunoexpression of tryptase, MMP-9 and MMP-13 (P = 0.002, P = < 0.001 and P < 0.001, respectively). In comparison with lesions with inflammatory infiltrates grades I and II, lesions with inflammatory infiltrate grade III had higher median percentages of MMP-13-positive cells (P = 0.003) and a tendency for higher expression of MMP-9 (P = 0.059). No significant difference was observed between the expression of the studied markers and epithelial thickness (P > 0.05). There were positive correlations between the number of tryptase-positive mast cells and the immunoexpression of MMP-9, as well as between the immunoexpression of MMP-9 and MMP-13. CONCLUSION: A larger number of tryptase-positive mast cells and greater enzymatic activity of MMP-9 and MMP-13 were found in PGs compared to RCs and RRCs. These findings are a characteristic of the dynamics of periapical diseases.

Biocompatibility and mineralized nodule formation of Neo MTA Plus and an experimental tricalcium silicate cement containing tantalum oxide.

AIM: To evaluate the biocompatibility and mineralized nodule formation of an experimental tricalcium silicate cement with tantalum oxide (TSC/Ta2 O5 ) as radiopacifier, Neo MTA Plus (Avalon Biomed Inc., Bradenton, FL, USA) and MTA (Angelus, Londrina, PR, Brazil) on human osteoblast-like cells (Saos-2). METHODOLOGY: Biocompatibility was evaluated by 3-(4,5-dimethyl-thiazoyl)-2,5-diphenyl-tetrazolium bromide (MTT) and neutral red (NR) assays, after exposure of Saos-2 to cement extracts at 1 : 1, 1 : 2, 1 : 4 and 1 : 8 dilutions for 24 h. Bioactivity was evaluated by alkaline phosphatase (ALP) activity, and calcium deposits were detected with alizarin red staining (ARS). Statistical analysis was performed with analysis of variance and Bonferroni or Tukey post-test (α = 0.05). RESULTS: The MTT assay revealed lower cytotoxicity for NEO and MTA (P < 0.05), and higher for TSC/Ta2 O5 at 1 : 1 and 1 : 2 dilutions when compared to serum-free medium - control (P > 0.05). At 1 : 4 dilution, the TSC/Ta2 O5 cytotoxicity was similar to the control (P > 0.05). At 1 : 8 dilution, cell viability was significantly greater than the control (P < 0.05). Saos-2 cell viability performed using the NR assay at all dilutions revealed no cytotoxic effect of MTA, NEO and TSC/Ta2 O5 . ALP activity at 1 and 3 days was similar to the control (P > 0.05). TSC/Ta2 O5 had significantly greater ALP activity at 7 days when compared with the control (P < 0.05). All materials induced the production of mineralized nodules, and NEO produced significantly more mineralized nodules than MTA and TSC/Ta2 O5 (P < 0.05). CONCLUSIONS: Neo MTA Plus and TSC/Ta2 O5 were biocompatible and induced ALP activity in Saos-2 cells. Both materials induced mineralized nodule formation by Saos-2 with Neo MTA Plus producing significantly more.