Application of a robust analytical method for quantifying progestins in environmental samples from three Portuguese Estuaries.

In the last years, progestins have raised special concerns for their documented negative effects on aquatic species, yet little is known about their environmental levels in surface waters and bioaccumulation in the trophic web. This study aimed to 1) adapt an extraction method for quantifying progestins in freeze-dried matrices, 2) validate the analytical procedure for three matrices: bivalve, polychaete, and crustacean, and 3) characterize levels of the four most prescribed synthetic progestins in key species across three Portuguese estuaries. Through the validated method, progestins were only quantifiable for the crustacean. Values were generally low, peaking with drospirenone values in Ria de Aveiro (1.33 ± 0.26 ng/g ww) and Tagus estuary (1.42 ± 0.55 ng/g ww), while Ria Formosa exhibited the lowest progestin concentrations (< 1 ng/g ww). This study enabled the development of a precise extraction and analytical method for quantifying steroid hormones in three distinct biological matrices.

Baseline progestins characterization in surface waters of three main Portuguese estuaries.

Synthetic progestins are micropollutants of special concern, due to their growing use in human and veterinary therapies and their risks to aquatic life. Currently, there is a lack of environmental information on these compounds, worldwide. The main objective of this work was to characterize the levels of the most consumed progestins in Portugal. For that, Ria de Aveiro, Tagus estuary and Ria Formosa were sampled in a temporal perspective to evaluate levels of drospirenone (DRO), desogestrel (DSG), gestodene (GST) and levonorgestrel (LNG). Drospirenone and desogestrel were the most abundant progestins. In the North of Portugal, DSG was the most abundant (Aveiro: 193.9 ng L-1 in summer), while DRO was more representative in the South (Tagus: 178.9 ng L-1; Formosa: 125.7 ng L-1) and also in summer. These spatial differences can be associated with the hydrodynamics of each estuarine system as well as the distinct population and tourist levels associated with each site.

Biological role of the bidirectional interaction between epithelial-mesenchymal transition and PD-L1 expression in head and neck squamous cell carcinomas: A systematic review.

BACKGROUND: Squamous cell carcinoma (SCC) is the most common head and neck malignant neoplasm. Despite progress in antineoplastic treatment for SCC, there are still high morbidity and mortality rates. Over the years, several tumor biomarkers have been suggested to predict the prognosis of patients with oral SCC. Studies point to a bidirectional association between the epithelial-mesenchymal transition (EMT) and the expression of PD-L1 with the aggressive biological behavior of the neoplastic cell. Thus, this systematic review aimed to explore the biological roles and mechanisms underlying the interaction between EMT and PD-L1 expression in head and neck SCC-derived cell lines. MATERIAL AND METHODS: An electronic search was performed in the PubMed/Medline, Web of Science, Science Direct, Scopus, Embase, and Cochrane Collaboration Library databases. Articles evaluating the in vitro relationship between EMT/PD-L1 interaction and the biological behavior of head and neck SCC cell lines were selected for this systematic review. The quality of evidence was assessed using the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) criteria. RESULTS: After applying the previously established inclusion/exclusion criteria, 9 articles were included in the qualitative synthesis. The present systematic review suggests the existence of a bidirectional interaction between EMT and PD-L1 expression, which is related to alterations in the cell cycle, proliferation, apoptosis, and cell survival, affecting the migration and invasion ability of tumor cells. CONCLUSIONS: Combined targeting of the two pathways may be potentially effective for immunotherapy in head and neck SCC.

Combined effects of climate change and environmentally relevant mixtures of endocrine disrupting compounds on the fitness and gonads' maturation dynamics of Nucella lapillus (Gastropoda).

Coastal areas are affected by multiple stressors like climate change and endocrine disruptors (EDCs). In the laboratory, we investigated the combined effects of increased temperature and EDCs (drospirenone and mercury) on the fitness and gonads' maturation dynamics of the marine gastropod Nucella lapillus for 21 days. Survival was negatively affected by all the stressors alone, while, in combination, a synergistic negative effect was observed. Both chemicals, as single factors, did not cause any effect on the maturation stage of ovaries and testis. However, in the presence of a higher temperature, it was clear a delay in the maturation stage of the ovaries, but not in the testis, suggesting a higher negative impact of the stressors in females than in males. In summary, drospirenone caused a low negative impact in aquatic species, like gastropods, but in combination with other EDCs and/or increased temperature can be a matter of concern.

Prognostic and clinicopathological significance of tumor-stroma ratio in head and neck squamous cell carcinoma: A systematic review.

BACKGROUND: Analysis of the tumor microenvironment has been proposed as a strategy for the treatment and prognosis of different neoplastic processes. A grading system based on the tumor-stroma ratio (TSR), which evaluates the proportion of stroma in relation to neoplastic parenchyma at the invasion front, has shown a strong prognostic value in different neoplastic processes. The aim of the present systematic review was to understand the role of the TSR in head and neck squamous cell carcinoma (HNSCC), evaluating its correlation with clinical and prognostic parameters. MATERIAL AND METHODS: An electronic search was performed in PubMed/Medline, Web of Science, Science Direct, Scopus, Embase, and the Cochrane Collaboration Library. Publications assessing the relationship between TSR and prognosis in cases of HNSCC were eligible. The quality of the studies was assessed independently by four evaluators using the Newcastle-Ottawa scale. RESULTS: After application of the previously es+lished inclusion/exclusion criteria, nine articles were included in the qualitative synthesis. With regards to quality on the Newcastle-Ottawa scale, an overall value of 4.55 was obtained. This systematic review demonstrated a strong association between TSR and prognosis in esophageal and oral squamous cell carcinomas. CONCLUSIONS: Histopathological analysis of the TSR can optimize the analysis of the prognosis of cases diagnosed with HNSSC. In addition, the TSR is a reliable and simple parameter that can be evaluated in hematoxylin/eosin-stained slides during routine laboratory examinations, showing high inter- and intraobserver agreement.

Effects of co-administered dexamethasone and nimesulide on pain, swelling, and trismus following third molar surgery: a randomized, triple-blind, controlled clinical trial.

This study aimed to determine the effect of the co-administration of dexamethasone 8mg and nimesulide 100mg given 1h before mandibular third molar surgery. A prospective, randomized, triple-blind, split-mouth clinical trial was developed at the study institution in Pernambuco, Brazil. A pilot study was first performed (95% confidence interval, 80% test power, and 5% error), and a sample of 40 patients aged between 18 and 40 years was selected. The patients were randomized and divided into two groups: dexamethasone+placebo and dexamethasone+nimesulide. The following parameters were evaluated: pain (visual analogue scale), total number of rescue analgesics taken, time taken to first rescue analgesic consumption, oedema, trismus, and patient satisfaction. The paired t-test and the Wilcoxon test were used to compare means. Statistically significant differences were found between the groups in pain values at 2, 4, and 12h postoperative, and in the total number of rescue analgesics and time taken to first rescue analgesic ingestion (P<0.05), with results in favour of dexamethasone+nimesulide administration. Oedema and trismus were similar in the two treatment groups and decreased over time postoperatively. The co-administration of dexamethasone and nimesulide reduces pain intensity and the need for rescue medication after third molar surgery.

Eficiência nutricional do Nitrogênio e produção de biomassa em Calendula officinalis L. (Asteraceae) em condições de casa de vegetação / Nitrogen nutritional efficiency and biomass production in Calendula officinalis L. (Asteraceae) under greenhouse conditions

O objetivo deste trabalho foi avaliar o efeito da aplicação de diferentes doses de nitrogênio no solo e de épocas de coleta sobre a eficiência nutricional do nitrogênio e a produção de biomassa em Calendula officinalis L. (Asteraceae) em condições de casa de vegetação. O substrato utilizado foi a camada de 0 - 20 cm de Latossolo Vermelho Distroférrico. O delineamento experimental foi inteiramente casualizado, com quatro repetições. Os tratamentos foram constituídos de quatro doses de N (0, 21, 42 e 84mg dm-3) e cinco épocas de colheita (15, 30, 45, 60 e 90 dias após o transplante). O teor de N na inflorescência de calêndula reduziu com as épocas de coleta, redução essa que ocorreu possivelmente devido à menor eficiência da absorção desse elemento pela planta e menor translocação do N para as inflorescências. Pode se concluir que as épocas de avaliação influenciaram na resposta da planta à eficiência de absorção (EFA), sendo a máxima EFA de N aos 51 dias após o transplante das mudas. Observou-se aumento na translocação de N da parte aérea (folhas e caules) para as inflorescências, tendo como consequência a redução do N na parte aérea. Houve incremento na produção de biomassa de calêndula em função das épocas de coleta , e do aumento das doses de N aplicadas no solo. A eficiência nutricional do N foi influenciada pelas épocas de avaliação e pelo teor de N presente no solo.

The aim of this study was to evaluate the effect of both the application of different nitrogen levels on the soil and the harvest periods on nitrogen nutritional efficiency and biomass production in Calendula officinalis L. (Asteraceae) under greenhouse conditions. The employed substrate was 0-20cm layer of Oxisol. Experimental design was completely randomized, with four replicates. Treatments consisted of four N levels (0, 21, 42 and 84 mg dm-3) and five harvest periods (15, 30, 45, 60 and 90 days after transplanting). N levels in Calendula officinalis inflorescences reduced with the harvest periods, and such a reduction was probably due to the lower efficiency of absorption of this element by the plant and to the lower N translocation to inflorescences. We can conclude that the evaluation periods influenced the response of plants to uptake efficiency (UE), and the maximal UE for N was found at 51 days after transplanting of seedlings. There was an increase in N translocation from the shoot (leaves and stems) to inflorescences, which has as consequence N reduction in the shoot. Calendula officinalis biomass production increased with the harvest periods and with the increase in the N levels applied on the soil. N nutritional efficiency was influenced by both the evaluation periods and the N levels present on the soil.

Identification of plant families associated with the predators Chrysoperla externa (Hagen) (Neuroptera: Chrysopidae) and Hippodamia convergens Guérin-Menéville (Coleoptera: Coccinelidae) using pollen grain as a natural marker.

The predators Hippodamia convergens Guérin-Menéville (Coleoptera: Coccinelidae) and Chrysoperla externa (Hagen) (Neuroptera: Chrysopidae), are frequently observed on vegetable crops, especially on tomato plants, as well as on flowers of several plants around crop fields. It is well known that when predators feed on pollen and nectar they can increase their longevity and reproductive capacity. The objective of this work was to identify plants that could be a pollen source for H. convergens and C. externa in order to develop strategies to attract and keep these predators in vegetable fields like the tomato crop. Adults of C. externa (53 individuals) and H. convergens (43 individuals) were collected in fields from 2004-2005 at Embrapa Hortaliças, Brasília, Federal District. The insects were processed by the acetolysis method and pollen from them was extracted and identified. A total of 11335 grains of pollen belonging to 21 families were extracted from C. externa. A total of 46 pollen grains belonging to ten families were extracted from H. convergens. The Poaceae family was the most abundant one for C. externa while Asteraceae was the commonest pollen for H. convergens. The importance of pollen from different plant species as a food resource for each predator species gives an indication of the importance of plant community structure inside and around crop fields for the establishment of these predator populations and to enhance conservation biological control.

Climate and host plant characteristics effects on lepidopteran caterpillar abundance on Miconia ferruginata DC and Miconia pohliana Cogn (Melastomataceae).

Folivore cerrado caterpillars are found on their host species in low frequencies, which vary between plants and throughout the year. We analysed the effects of climatic variation and of the characteristics of two host species (Miconia, Melastomataceae) on caterpillar abundance. The work was carried out in a cerrado sensu stricto area in Brasília, Federal District, Brazil, from May 1998 to September 1999. One hundred plants of each species were inspected monthly. All caterpillars found were collected and raised in the laboratory with Miconia leaves as food sources. Climatic data for the study area and plant characteristics such as local density, spatial distribution, foliar phenology, leaf pilosity and nutritional content were used to test the effects on caterpillar abundance. Less than 10% of the plants had larvae. Caterpillars were more frequent and abundant in M. pohliana, which was also present at higher densities than M. ferruginata. Low caterpillar frequency in host plants and an abundance peak during the dry season are consistent patterns for different cerrado Lepidoptera larvae. This abundance peak, however, is not coincident with higher leaf production in host species. We suggest that M. pohliana's higher local density and better leaf digestibility can account for the higher abundance of caterpillars in this species.

Survey of owned feline and canine populations in apartments from a neighbourhood in Curitiba, Brazil.

Animal population estimates are essential for public health services to ensure the success of zoonoses control programmes. Canine and feline populations vary among different regions mainly because of local human income status and type of human residence. Accordingly, the present study estimated the pet population size living in apartments of a vertical neighbourhood in Curitiba, Brazil. We chose a neighbourhood with a predominance of apartment buildings. All apartment buildings were visited, and questionnaires were completed by doormen or residents. Data were obtained from 120 of 173 apartment buildings. Survey questions included the number of apartments, residents, dogs and cats. Two thousand nine hundred and sixty six apartments with a total of 7429 residents were surveyed. The number of dogs and cats was 569 and 86 respectively. Thus, the human:dog and human:cat ratios were 13.05:1 and 86.38:1. These ratios were higher than those observed in other neighbourhoods in Curitiba. The present study indicates that the number of pets from apartments may be different from houses, and different among distinct areas within the same city.

Serum cardiac troponin I concentration in dogs with ehrlichiosis.

BACKGROUND: Ehrlichiosis is a multisystemic disease with the potential to cause cardiomyocyte injury in naturally infected dogs. HYPOTHESIS: Myocardial injury occurs in dogs infected with Ehrlichia canis. ANIMALS: One-hundred and ninety-four dogs from Brazil with clinical and laboratory abnormalities indicative of ehrlichiosis. Sixteen healthy dogs served as controls. METHODS: Electrocardiogram, echocardiogram, noninvasive blood pressure measurement, and serum cardiac troponin I (cTnI) concentrations were evaluated. Serologic assays and PCR determined the exposure and infection status for E. canis, Anaplasma spp., Babesia canis vogeli, Bartonella spp., Borrelia burgdorferi, Dirofilaria immitis, Ehrlichia chaffeensis, Ehrlichia ewingii, Leishmania chagasi, and spotted-fever group Rickettsia. Dogs were assigned to groups according to PCR status: E. canis infected, infected with other vector-borne organisms, sick dogs lacking PCR evidence for infection, and healthy controls. RESULTS: E. canis-infected dogs had higher serum cTnI concentrations than controls (median: 0.04 ng/dL; range 0.04-9.12 ng/dL; control median: 0.04 ng/dL; range: 0.04-0.10 ng/dL; P= .012), and acute E. canis infection was associated with myocardial injury (odds ratio [OR]: 2.67, confidence interval [CI] 95%: 1.12-6.40, P= .027). Severity of anemia was correlated with increased risk of cardiomyocyte damage (r= 0.84, P< .001). Dogs with clinical signs of systemic inflammatory response syndrome (SIRS) were at higher risk for myocardial injury than were other sick dogs (OR: 2.55, CI 95%: 1.31-4.95, P= .005). CONCLUSIONS AND CLINICAL IMPORTANCE: Acute infection with E. canis is a risk factor for myocardial injury in naturally infected Brazilian dogs. Severity of anemia and SIRS might contribute to the pathophysiology of myocardial damage.

Host specificity of Lepidoptera in tropical and temperate forests.

For numerous taxa, species richness is much higher in tropical than in temperate zone habitats. A major challenge in community ecology and evolutionary biogeography is to reveal the mechanisms underlying these differences. For herbivorous insects, one such mechanism leading to an increased number of species in a given locale could be increased ecological specialization, resulting in a greater proportion of insect species occupying narrow niches within a community. We tested this hypothesis by comparing host specialization in larval Lepidoptera (moths and butterflies) at eight different New World forest sites ranging in latitude from 15 degrees S to 55 degrees N. Here we show that larval diets of tropical Lepidoptera are more specialized than those of their temperate forest counterparts: tropical species on average feed on fewer plant species, genera and families than do temperate caterpillars. This result holds true whether calculated per lepidopteran family or for a caterpillar assemblage as a whole. As a result, there is greater turnover in caterpillar species composition (greater beta diversity) between tree species in tropical faunas than in temperate faunas. We suggest that greater specialization in tropical faunas is the result of differences in trophic interactions; for example, there are more distinct plant secondary chemical profiles from one tree species to the next in tropical forests than in temperate forests as well as more diverse and chronic pressures from natural enemy communities.

Detection of canine distemper virus by reverse transcriptase-polymerase chain reaction in the urine of dogs with clinical signs of distemper encephalitis.

In a prospective study we evaluated the use of the reverse transcriptase-polymerase chain reaction (RT-PCR) in urine samples to diagnose canine distemper virus in dogs with progressive neurological disease. A fragment of the nucleoprotein gene of canine distemper virus was amplified from the urine of 22 distemper dogs. The body fluids and leukocytes of 12 asymptomatic dogs were RT-PCR negative. RT-PCR of urine samples was more sensitive than serum and leukocytes and at least as sensitive as cerebrospinal fluid to screen for distemper in dogs with neurological signs and extraneural systemic signs.

Climatic unpredictability and parasitism of caterpillars: implications of global warming.

Insect outbreaks are expected to increase in frequency and intensity with projected changes in global climate through direct effects of climate change on insect populations and through disruption of community interactions. Although there is much concern about mean changes in global climate, the impact of climatic variability itself on species interactions has been little explored. Here, we compare caterpillar-parasitoid interactions across a broad gradient of climatic variability and find that the combined data in 15 geographically dispersed databases show a decrease in levels of parasitism as climatic variability increases. The dominant contribution to this pattern by relatively specialized parasitoid wasps suggests that climatic variability impairs the ability of parasitoids to track host populations. Given the important role of parasitoids in regulating insect herbivore populations in natural and managed systems, we predict an increase in the frequency and intensity of herbivore outbreaks through a disruption of enemy-herbivore dynamics as climates become more variable.

Tratamento de insuficiência cardíaca com benazepril em cães com cardiomiopatia dilatada e endocardiose / Treatment of congestive heart failure with benazepril in dogs with dilated cardiomyopathy and endocardiosis

Foram avaliados os efeitos clínicos do benazepril, um inibidor da enzima de conversão da angiotensina de ação prolongada, em cães com insuficiência cardíaca congestiva (ICC) secundária à endocardiose de mitral ou cardiomiopatia dilatada. O medicamento foi administrado na dose de 0,25 a 0,5mg/kg/dia. Diuréticos, digitálicos e fármacos antiarrítmicos froam usados de acordo com a necessidade de cada paciente. Exames físico, radiográfico e eletrocardiográfico foram realizados nos dias 0, 7, 28 e 56. A gasometria arterial e a bioquímica sérica foram avaliadas nos dias 0 e 56. Os sinais de dispnéia e o estado geral dos pacientes melhoraram em todos os cães após o início do tratamento. Houve diminuição na frequência da tosse e não houve alterações no eletrocardiograma, exceto pela diminuição na amplitude e na duração da onda P. Nenhum efeito colateral foi observado. Conclui-se que o benazepril é um inibidor da enzima de conversão da angiotensina, eficaz e bem tolerado no tratamento da ICC no cão.

Immunohistochemistry of atrial and brain natriuretic peptides in control cats and cats with hypertrophic cardiomyopathy.

Atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) are cardiac hormones involved in electrolyte and fluid homeostasis. Our laboratory has investigated the use of ANP and BNP as diagnostic markers of cardiac disease in cats. We hypothesize that the cardiac distribution of ANP and BNP increases in cats with hypertrophic cardiomyopathy (HCM). Accordingly, we evaluated the immunohistochemical distribution of ANP and BNP in hearts of four cats with naturally occurring HCM relative to five healthy controls. Indirect immunoperoxidase was performed with polyclonal immunoglobulin G against feline ANP (1-28) and proBNP (43-56). In control cats, ANP and BNP immunoreactivity was restricted to the atria. Staining for both peptides was most intense adjacent to the endocardial surface. Auricles stained more diffusely than atria for both peptides. The interstitial capillaries and nerve fibers within the heart were positive only for BNP. Atrial immunoreactivity for ANP and BNP was more diffuse and had a less distinctly layered pattern in HCM than in control cats. Ventricular cardiomyocytes of HCM cats were negative for ANP but stained lightly and diffusely for BNP. The capillaries and nerve fibers remained positive for BNP. We conclude that in cats with HCM, the cardiac distribution of ANP and BNP is more diffuse in the atria and that novel expression of BNP in the ventricular cardiomyocytes occurs.

Effect of ascorbic acid on the stability of beta-carotene and capsanthin in paprika (Capsicum annuum) powder.

The effect of ascorbic acid, light, and storage on the stability of the pigments beta-carotene and capsanthin in red pepper (Capsicum annuum) powder has been elucidated by determining the amount of pigment in samples treated by various concentrations of ascorbic acid. Determination of pigment concentration has been performed after different storage times using high-performance liquid chromatography. The dependence of the concentration of pigments on the concentration of ascorbic acid, presence of light and the storage time has been assessed by stepwise regression analysis. The concentration of pigments decreased at longer storage time and increased at higher concentration of ascorbic acid, beta-carotene being more sensitive towards storage time and concentration of ascorbic acid than capsanthin. Interaction between the effects of light and storage time, and light and concentration of ascorbic acid has been established.

The mixed acid-base disturbances of severe canine babesiosis.

Thirty-four dogs suffering from severe babesiosis caused by Babesia canis rossi were included in this study to evaluate acid-base imbalances with the quantitative clinical approach proposed by Stewart. All but 3 dogs were severely anemic (hematocrit <12%). Arterial pH varied from severe acidemia to alkalemia. Most animals (31 of 34; 91%) had inappropriate hypocapnia with the partial pressure of CO2 < 10 mm Hg in 12 of 34 dogs (35%). All dogs had a negative base excess (BE; mean of - 16.5 mEq/L) and it was below the lower normal limit in 25. Hypoxemia was present in 3 dogs. Most dogs (28 of 34; 82%) were hyperlactatemic. Seventy percent of dogs (23 of 33) were hypoalbuminemic. Anion gap (AG) was widely distributed, being high in 15, low in 12, and normal in 6 of the 33 dogs. The strong ion difference (SID; difference between the sodium and chloride concentrations) was low in 20 of 33 dogs, chiefly because of hyperchloremia. Dilutional acidosis was present in 23 of 34 dogs. Hypoalbuminemic alkalosis was present in all dogs. Increase in unmeasured strong anions resulted in a negative BE in all dogs. Concurrent metabolic acidosis and respiratory alkalosis was identified in 31 of 34 dogs. A high AG metabolic acidosis was present in 15 of 33 dogs. The lack of an AG increase in the remaining dogs was attributed to concurrent hypoalbuminemia, which is common in this disease. Significant contributors to BE were the SID, free water abnormalities, and AG (all with P < .01). Mixed metabolic and respiratory acid-base imbalances are common in severe canine babesiosis, and resemble imbalances described in canine endotoxemia and human malaria.

Canine paracoccidioidomycosis: a seroepidemiologic study.

Sera from 305 dogs were analyzed by enzyme-linked immunosorbent assay (ELISA) to determine presence of the antibody anti-gp43, which reacts to a specific antigen of Paracoccidioides brasiliensis. The dogs were divided into three groups according to their origin: urban dogs (animals with little or no contact with rural areas); suburban dogs (from the urban outskirts); and rural dogs. There was a significant difference between groups (P < 0.05). Rural dogs reacted positively in 89.5% of cases, followed by suburban (48.8%) and urban dogs (14.8%). There were no differences between male and female dogs. In an attempt to verify the feasibility of skin testing with gp43 to determine sensitization against P. brasiliensis in dogs, suburban (n = 61) and rural (n = 21) dogs were tested, showing positivity of 13.1 and 38.1%, respectively. Six dogs that had higher ELISA titers and also showed strong reactions in skin testing were killed in an attempt to isolate P. brasiliensis. The fungus was not detected by culture or histopathological analysis in these dogs, suggesting that dogs have a natural resistance or that they encounter an inoculum level that is insufficient to cause disease. These results indicate that ELISA and skin testing can be useful in the epidemiological study of paracoccidioidomycosis in dogs and that encounter with the fungus in nature is a frequent event.