Immunohistochemical evaluation of e-cadherin, Ki-67 and PCNA in canine mammary neoplasias: correlation of prognostic factors and clinical outcome

E-cadherin is a cell-cell adhesion molecule and low e-cadherin expression is related to invasiveness and may indicate a bad prognosis in mammary neoplasms. The expression of cell proliferation markers PCNA and especially Ki-67, has also proved to have a strong prognostic value in this tumor class. The expression of these markers was related to the clinical-pathological characteristics of 73 surgically removed mammary tumors in female dogs by immunohistochemistry. There was no statistical correlation between these markers and death by neoplasm, survival time and disease-free interval. However, the loss of e-cadherin expression and marked Ki-67 expression (p=0.016) were considered statistically significant for the diagnosis (p=0.032). When evaluated as independent factors, there was evidence of the relationship between the loss of e-cadherin expression and high PCNA expression with changes in the body status (divided into obese, normal and cachectic) of female dogs (p=0.030); there was also evidence of the relationship between pseudopregnancy and e-cadherin alone (p=0.021) and for ulceration and PCNA alone (p=0.035). The significant correlation between the markers expression and these well known prognostic factors used individually or in combination suggests their prognostic value in canine mammary tumors.

Goniometria dos membros pélvicos de pacas (Cuniculus paca) criadas em cativeiro

O trabalho teve por objetivo avaliar por meio de estudos goniométricos os membros pélvicos de pacas criadas em cativeiro. Foram utilizadas oito pacas (Cuniculus paca), sendo três fêmeas e cinco machos, com peso entre 6,0 kg e 8,2 kg, nos quais foi aferido o movimento de extensão e flexão das articulações coxofemoral, joelho e tarsocrural utilizando-se um goniômetro de plástico. Foram realizadas duas aferições para cada membro pelo mesmo avaliador, com intervalo de uma semana entre cada aferição, com os animais sob anestesia dissociativa. Não houve diferença estatística nas aferições dos ângulos entres os momentos. As variações entre as medidas de flexão e extensão, efetuadas em cada momento pelo mesmo avaliador, foi entre 1º e 5º para cada membro. Os valores goniométricos médios foram: flexão de 43,96º±7,62 e extensão de 118,31º±9,79 para a articulação coxofemoral; flexão de 54,25º±10,24 e extensão 131,53º±7,89 para o joelho; flexão de 45,94º±7,16 e extensão de 145,03º±5,06 para a articulação tarsocrural. Sendo assim, observou-se que a amplitude de movimento das pacas é maior na articulação tarsocrural, seguida respectivamente do joelho e da articulação coxofemoral.

Manejo de convolvuláceas em dois cultivares de soja semeada diretamente sob palha residual de cana crua

A interferência de convolvuláceas na produtividade dos cultivares de soja (Coodetec 206 e Monsoy 6101) semeados diretamente sob a palha residual (12,8 t ha-1) de cana crua foi estudada em relação aos manejos com o herbicida diclosulam (0,035 kg ha-1) em mistura em tanque com glyphosate (1,440 kg ha-1), aplicados na operação de dessecação da cana-de-açúcar, diclosulam isolado (0,035 kg ha-1), em pré-emergência da cultura da soja, e fomesafen (0,250 kg ha¹), em pós-emergência. Para isso, utilizou-se delineamento experimental de blocos casualizados, num esquema fatorial 2x5, com oito repetições. Observou-se pelos resultados que a utilização do herbicida diclosulam isoladamente e em mistura com o glyphosate promoveu controle eficaz das plantas daninhas. A infestação das cordas-de-viola não foi capaz de interferir negativamente nas características de crescimento das plantas, havendo apenas diferenças estatísticas entre os cultivares. As características de produção também não sofreram interferências negativas, exceto quanto à massa seca de 100 sementes no cultivar Monsoy 6101. Este cultivar Monsoy 6101 apresentou-se mais adequado para utilização dentro do período de renovação dos canaviais, por atingir o estádio R8 (colheita) nove dias antes.

Fitotoxicidade de herbicidas a diferentes cultivares de milho-pipoca

Com o objetivo de avaliar a seletividade de diferentes herbicidas a dez genótipos de milho-pipoca em Campos dos Goytacazes-RJ, realizou-se a aplicação de herbicidas em dosagem máxima recomendada (atrazine + S-metolachlor + extravon - 1,665 + 1,035 kg ha-1 i.a. + 0,1%; foramsulfuron + iodosulfuron + hoefix - 45,0 + 3,0 g ha-1 i.a. + 0,5%; mesotrione + óleo mineral - 192 g ha-1 + 0,5%; tembotrione + óleo mineral - 240 mL ha-1 + 0,5%) e um tratamento sem aplicação de herbicida, em esquema fatorial 5 x 10 x 5. O experimento foi conduzido no delineamento em blocos casualizados, com quatro repetições. Aos 2, 4, 8, 12 e 20 dias após aplicação (DAA) foram avaliados sintomas visuais de fitotoxicidade. Aos 36 DAA foram avaliados a altura das plantas e o diâmetro do caule, e aos 39 DAA, a área foliar e a massa seca da parte aérea. Entre os tratamentos, os mais seletivos às variedades testadas foram os herbicidas atrazine + S-metolachlor, aplicados em pré-emergência; no entanto, atrazine + S-metolachlor, mesotrione e foramsulfuron + iodosulfuron, aplicados em pós-emergência, causaram elevados níveis de fitotoxicidade às plantas de milho-pipoca, sendo as variedades Beija-Flor, Pr-023, SE-013, Angela, PA-038 e UFV extremamente sensíveis a esses produtos.

Residual activity of herbicides used in soybean agriculture on grain sorghum crop succession

The sorghum is a kind of prominence before the cultures used in succession in the Brazil. However, little information concerning the effects of residual activity of herbicides on the crop in this region are known. The objective of this study was to evaluate the residual activity of herbicides used in weed management in soybeans as well as check their effects on grain sorghum grown in succession. For the field experiment, we used a randomized block design with four replications. Eight herbicide treatments were evaluated: imazaquin (0,161 kg ha-1), diclosulam (0,035 kg ha-1), sulfentrazone (0,600 kg ha-1) and flumioxazin (0,05 kg ha-1) in applications made before emergency and chlorimuron-ethyl (0,015 kg ha-1), imazethapyr (0,060 kg ha-1), imazethapyr (0,100 kg ha-1) and fomesafen (0,250 kg ha-1) applied post-emergence soybean (V3 stadium, 18 DAE), and a control without herbicide application. The grain sorghum (cv. AG-1040) was sown after the harvest of soybeans. The residual activity of these herbicides was determined by bioassay, using the same sorghum cultivars evaluated in the field during the period from 0 to 200days after application the treatments. The sorghum crop showed high sensitivity to residual activity of the herbicide sulfentrazone, diclosulam and imazethapyr dose of 0,100 kg ha-1, even when grown after soybean harvest. Furthermore, the residual activity of sulfentrazone exceeded the range of assessment of bioassay, and more than 200 days.

Atividade residual de herbicidas pré-emergentes aplicados na cultura da soja sobre o milheto cultivado em sucessão

O objetivo deste estudo foi avaliar a atividade residual de herbicidas utilizados no manejo de plantas daninhas em pré-emergência da cultura da soja, bem como verificar seus efeitos sobre o milheto cultivado em sucessão. Utilizaram-se parcelas de 80 m², em delineamento de blocos casualizados, com quatro repetições, em esquema de parcelas subdivididas 5 x 4, correspondendo à aplicação dos herbicidas imazaquin (0,160 kg ha-1), diclosulam (0,035 kg ha-1), sulfentrazone (0,600 kg ha-1) e flumioxazin (0,050 kg ha-1) e uma testemunha, aplicados logo após a semeadura do cultivar de soja Msoy-6101. Nas subparcelas, realizou-se a semeadura do milheto, cultivar ADR-7010, em quatro períodos, correspondendo a 0, 40, 80 e 120 dias após a aplicação dos herbicidas (DAA). Durante a condução do ensaio, foram determinados os níveis de intoxicação, estande, altura e massa seca da parte aérea das plantas de milheto. No final do ciclo foi avaliado o rendimento de grãos da cultura. O híbrido de milheto ADR-7010 apresentou elevada sensibilidade com relação à atividade residual dos herbicidas sulfentrazone, diclosulam e imazaquin quando cultivado logo após a aplicação destes. A bioatividade dos herbicidas imazaquin, diclosulam e flumioxazin não foi suficiente para alterar o rendimento de grãos do milheto cultivado em sucessão à soja (120 DAA), mostrando que esse intervalo de tempo é suficiente para dissipação desses herbicidas. Dos herbicidas pré-emergentes avaliados, o sulfentrazone apresentou maior atividade residual, influenciado negativamente o rendimento da cultura durante o intervalo de tempo estudado.

Seleção de espécies de adubos verdes visando à fitorremediação de diclosulam

Espécies de adubos verdes tolerantes ao herbicida diclosulam podem ser utilizadas em rotação de culturas para diminuir o efeito fitotóxico subsequente desse herbicida em plantas sensíveis, como o girassol ou milho. Este estudo teve como objetivo avaliar a tolerância de adubos verdes ao diclosulam e a capacidade dessas plantas em diminuir o efeito fitotóxico do herbicida no bioindicador Helianthus annuus. Foram avaliadas, em casa de vegetação, três doses do herbicida diclosulam (0, 0,035 e 0,070 kg i.a. ha-1) em pré-emergência dos adubos verdes Dolichos lablab, Cajanus cajan, Canavalia ensiformis, Crotalaria juncea, C. breviflora, C. spectabilis, Mucuna deeringiana, M. cinerea, M. aterrima, Lupinus albus, Helianthus annuus, Pennisetum glaucum, Avena strigosa, Raphanus sativus e Calopogonium muconoides. Entre estas espécies, C. cajan, C. ensiformis, M. cinerea e M. aterrima foram selecionadas como as mais tolerantes, sendo avaliadas no campo com o herbicida diclosulam nas doses de 0, 0,035 e 0,070 kg i.a. ha¹, em esquema fatorial 4 x 3 e quatro repetições. A parte aérea dessas plantas foi coletada após 60 dias da emergência, sendo semeado H. annuus como bioindicador do herbicida diclosulam. Os resultados evidenciaram C. cajan como a espécie mais promissora em diminuir o efeito fitotóxico do diclosulam em culturas agrícolas sensíveis.

Estrutura genética populacional de Copaifera langsdorffii Desf. (Leguminosae - Caesalpinioideae) em fragmentos florestais no Pontal do Paranapanema, SP, Brasil

O Pontal do Paranapanema é a região mais recentemente desflorestada do Estado de São Paulo. Os efeitos da fragmentação florestal na estrutura genética de Copaifera langsdorffii Desf. foram avaliados por meio da genotipagem, com seis locos microssatélites, de pelo menos 30 indivíduos regenerantes e 30 adultos em três áreas do Pontal do Paranapanema. A amostragem compreendeu dois fragmentos localizados em assentamentos rurais, Tucano (800 ha) e Madre Cristina (300 ha); e uma população localizada em floresta contínua, no Parque Estadual do Morro do Diabo (35.000 ha). Níveis elevados de diversidade genética foram detectados tanto nos regenerantes como nos adultos. A diversidade gênica (He) variou de 0,718 a 0,835 e o número médio de alelos por loco (Â) variou de 8,67 a 11,83, mostrando que a fragmentação ainda não causou perda de alelos. As estimativas elevadas dos índices de fixação (0,183 a 0,387) foram conseqüentes da ocorrência de alelos nulos em alguns locos e de endogamia nas populações. A divergência genética foi pequena entre os adultos (R ST = 0,035), mas aumentou entre os regenerantes (R ST = 0,075). Como esperado, a fragmentação florestal está causando redução do fluxo gênico interpopulacional, especialmente em relação ao fragmento mais distante da floresta contínua (Tucano - cerca de 8 km). Uma alternativa para promover o fluxo gênico entre os remanescentes florestais do Pontal do Paranapanema seria o estabelecimento de sistemas agroflorestais pelos agricultores assentados, os quais atuariam como trampolins ecológicos, favorecendo a dispersão de pólen e sementes entre fragmentos.

Characterization of six rat strains (Rattus norvegicus) by mitochondrial DNA restriction fragment length polymorphism

Restriction fragment length polymorphism (RFLP) was used to examine the extent of mtDNA polymorphism among six strains of rats (Rattus norvegicus) - Wistar, Wistar Munich, Brown Norway, Wistar Kyoto, SHR and SHR-SP. A survey of 26 restriction enzymes has revealed a low level of genetic divergence among strains. The sites of cleavage by EcoRI, NcoI and XmnI were shown to be polymorphic. The use of these three enzymes allows the 6 strains to be classified into 4 haplotypes and identifies specific markers for each one. The percentage of sequence divergence among all pairs of haplotypes ranged from 0.035 to 0.33%, which is the result of a severe population constriction undergone by the strains. These haplotypes are easily demonstrable and therefore RFLP analysis can be employed for genetic monitoring of rats within animal facilities or among different laboratories.

Effects of mercury on the arterial blood pressure of anesthetized rats

The available data suggests that hypotension caused by Hg2+ administration may be produced by a reduction of cardiac contractility or by cholinergic mechanisms. The hemodynamic effects of an intravenous injection of HgCl2 (5 mg/kg) were studied in anesthetized rats (N = 12) by monitoring left and right ventricular (LV and RV) systolic and diastolic pressures for 120 min. After HgCl2 administration the LV systolic pressure decreased only after 40 min (99 ± 3.3 to 85 ± 8.8 mmHg at 80 min). However, RV systolic pressure increased, initially slowly but faster after 30 min (25 ± 1.8 to 42 ± 1.6 mmHg at 80 min). Both right and left diastolic pressures increased after HgCl2 treatment, suggesting the development of diastolic ventricular dysfunction. Since HgCl2 could be increasing pulmonary vascular resistance, isolated lungs (N = 10) were perfused for 80 min with Krebs solution (continuous flow of 10 ml/min) containing or not 5 µM HgCl2. A continuous increase in pulmonary vascular resistance was observed, suggesting the direct effect of Hg2+ on the pulmonary vessels (12 ± 0.4 to 29 ± 3.2 mmHg at 30 min). To examine the interactions of Hg2+ and changes in cholinergic activity we analyzed the effects of acetylcholine (Ach) on mean arterial blood pressure (ABP) in anesthetized rats (N = 9) before and after Hg2+ treatment (5 mg/kg). Using the same amount and route used to study the hemodynamic effects we also examined the effects of Hg2+ administration on heart and plasma cholinesterase activity (N = 10). The in vivo hypotensive response to Ach (0.035 to 10.5 µg) was reduced after Hg2+ treatment. Cholinesterase activity (µM h-1 mg protein-1) increased in heart and plasma (32 and 65%, respectively) after Hg2+ treatment. In conclusion, the reduction in ABP produced by Hg2+ is not dependent on a putative increase in cholinergic activity. HgCl2 mainly affects cardiac function. The increased pulmonary vascular resistance and cardiac failure due to diastolic dysfunction of both ventricles are factors that might contribute to the reduction of cardiac output and the fall in arterial pressure.

Use of neuron-specific enolase for assessing the severity and outcome of neurological disorders in patients

Neuron-specific enolase (NSE) is a glycolytic enzyme present almost exclusively in neurons and neuroendocrine cells. NSE levels in cerebrospinal fluid (CSF) are assumed to be useful to estimate neuronal injury and clinical outcome of patients with serious clinical manifestations such as those observed in stroke, head injury, anoxic encephalopathy, encephalitis, brain metastasis, and status epilepticus. We compared levels of NSE in serum (sNSE) and in CSF (cNSE) among four groups: patients with meningitis (N = 11), patients with encephalic injuries associated with impairment of consciousness (ENC, N = 7), patients with neurocysticercosis (N = 25), and normal subjects (N = 8). Albumin was determined in serum and CSF samples, and the albumin quotient was used to estimate blood-brain barrier permeability. The Glasgow Coma Scale score was calculated at the time of lumbar puncture and the Glasgow Outcome Scale (GOS) score was calculated at the time of patient discharge or death. The ENC group had significantly higher cNSE (P = 0.01) and albumin quotient (P = 0.005), but not sNSE (P = 0.14), levels than the other groups (Kruskal-Wallis test). Patients with lower GOS scores had higher cNSE levels (P = 0.035) than patients with favorable outcomes. Our findings indicate that sNSE is not sensitive enough to detect neuronal damage, but cNSE seems to be reliable for assessing patients with considerable neurological insult and cases with adverse outcome. However, one should be cautious about estimating the severity of neurological status as well as outcome based exclusively on cNSE in a single patient.

Correlation between serum E-selectin levels and panoramic nailfold capillaroscopy in systemic sclerosis

E-selectin is expressed by the activated endothelium and its plasma levels are increased in patients with systemic sclerosis. Eighteen patients fulfilling the American Rheumatism Association criteria for systemic sclerosis, 15 females and 3 males, 42-70 years old, 9 with diffuse and 9 with limited forms, were sequentially recruited for this study. Serum E-selectin levels were determined by commercially available ELISA and their association with nailfold capillaroscopic abnormalities was investigated. Nailfold capillaries were analyzed by 16X magnification wide-field capillaroscopy. Two parameters on capillaroscopy were used to correlate to serum E-selectin: deletion and ectasia. Data were analyzed statistically by the Student t-test and Spearman correlation. Two-tailed P values below 0.05 were considered significant. E-selectin range was 38 to 200 ng/ml (80 ± 39.94). There was a correlation between serum E-selectin levels and the deletion capillaroscopic score (r = 0.50, P < 0.035). This correlation was even stronger within the first 48 months of diagnosis (r = 0.63, P < 0.048). On the other hand, no association was observed between selectin and ectasia. Patients with diffuse disease presented higher serum E-selectin levels than patients with limited disease, although the difference was not statistically significant (96.44 ± 48.04 vs 63.56 ± 21.77 ng/dl; P = 0.08). The present study is the first showing a correlation between soluble serum E-selectin levels and alterations in capillaroscopy. The stronger correlation of deletion score in capillaroscopy in early disease suggests that serum E-selectin levels might be a useful biochemical marker of disease activity in systemic sclerosis.

Indoor NO2 air pollution and lung function of professional cooks

Studies of cooking-generated NO2 effects are rare in occupational epidemiology. In the present study, we evaluated the lung function of professional cooks exposed to NO2 in hospital kitchens. We performed spirometry in 37 cooks working in four hospital kitchens and estimated the predicted FVC, FEV1 and FEF25-75, based on age, sex, race, weight, and height, according to Knudson standards. NO2 measurements were obtained for 4 consecutive days during 4 different periods at 20-day intervals in each kitchen. Measurements were performed inside and outside the kitchens, simultaneously using Palm diffusion tubes. A time/exposure indicator was defined as representative of the cumulative exposure of each cook. No statistically significant effect of NO2 exposure on FVC was found. Each year of work as a cook corresponded to a decrease in predicted FEV1 of 2.5% (P = 0.046) for the group as a whole. When smoking status and asthma were included in the analysis the effect of time/exposure decreased about 10% and lost statistical significance. On predicted FEF25-75, a decrease of 3.5% (P = 0.035) was observed for the same group and the inclusion of controllers for smoking status and asthma did not affect the effects of time/exposure on pulmonary function parameter. After a 10-year period of work as cooks the participants of the study may present decreases in both predicted FEV1 and FEF25-75 that can reach 20 and 30%, respectively. The present study showed small but statistically significant adverse effects of gas stove exposure on the lung function of professional cooks.

Imbalance of naive and memory T lymphocytes with sustained high cellular activation during the first year of life from uninfected children born to HIV-1-infected mothers on HAART

The immune consequences of in utero HIV exposure to uninfected children whose mothers were submitted to highly active antiretroviral therapy (HAART) during gestation are not well defined. We evaluated 45 HIV-exposed uninfected (ENI) neonates and 45 healthy unexposed control (CT) neonates. All HIV-infected mothers received HAART during pregnancy, and the viral load at delivery was <50 copies/mL for 56.8%. Twenty-three ENI neonates were further evaluated after 12 months and compared to 23 unexposed healthy age-matched infants. Immunophenotyping was performed by flow cytometry in cord and peripheral blood. Cord blood lymphocyte numbers did not differ between groups. However, ENI neonates had a lower percentage of naive T cells than CT neonates (CD4+, 76.6 vs 83.1%, P < 0.001; CD8+, 70.9 vs 79.6%, P = 0.003) and higher percentages of central memory T cells than CT neonates (CD4+, 13.9 vs 8.7%, P < 0.001; CD8+, 8.6 vs 4.8%, P = 0.001). CD38 mean fluorescence intensity of T cells was higher in ENI neonates (CD4+, 62.2 vs 52.1, P = 0.007; CD8+, 47.7 vs 35.3, P < 0.001). At 12 months, ENI infants still had higher mean fluorescence intensity of CD38 on T cells (CD4+, 34.2 vs 23.3, P < 0.001; CD8+, 26.8 vs 19.4, P = 0.035). Despite effective maternal virologic control at delivery, HIV-exposed uninfected children were born with lower levels of naive T cells. Immune activation was present at birth and remained until at least 12 months of age, suggesting that in utero exposure to HIV causes subtle immune abnormalities.

Mechanical cardiac remodeling and new-onset atrial fibrillation in long-term follow-up of subjects with chronic Chagas' disease

Atrial fibrillation (AF) affects subjects with Chagas' disease and is an indicator of poor prognosis. We investigated clinical, echocardiographic and electrocardiographic variables of Chagas' disease in a long-term longitudinal study as predictors of a new-onset AF episode lasting >24 h, nonfatal embolic stroke and cardiac death. Fifty adult outpatients (34 to 74 years old, 62% females) staged according to the Los Andes classification were enrolled. During a follow-up of (mean ± SD) 84.2 ± 39.0 months, 9 subjects developed AF (incidence: 3.3 ± 1.0%/year), 5 had nonfatal stroke (incidence: 1.3 ± 1.0%/year), and nine died (mortality rate: 2.3 ± 0.8%/year). The progression rate of left ventricular mass and left ventricular ejection fraction was significantly greater in subjects who experienced AF (16.4 ± 20.0 g/year and -8.6 ± 7.6%/year, respectively) than in those who did not (8.2 ± 8.4 g/year; P = 0.03, and -3.0 ± 2.5%/year; P = 0.04, respectively). In univariate analysis, left atrial diameter ≥3.2 cm (P = 0.002), pulmonary arterial hypertension (P = 0.035), frequent premature supraventricular and ventricular contraction counts/24 h (P = 0.005 and P = 0.007, respectively), ventricular couplets/24 h (P = 0.002), and ventricular tachycardia (P = 0.004) were long-term predictors of AF. P-wave signal-averaged ECG revealed a limited long-term predictive value for AF. In chronic Chagas' disease, large left atrial diameter, pulmonary arterial hypertension, frequent supraventricular and ventricular premature beats, and ventricular tachycardia are long-term predictors of AF. The rate of left ventricular mass enlargement and systolic function deterioration impact AF incidence in this population.