Metallopeptidase inhibitors arrest vital biological processes in the fungal pathogen Scedosporium apiospermum

P>Scedosporium apiospermum is an emerging agent of opportunistic mycoses in humans. Previously, we showed that mycelia of S. apiospermum secreted metallopeptidases which were directly linked to the destruction of key host proteins. In this study, we analysed the effect of metallopeptidase inhibitors on S. apiospermum development. As germination of inhaled conidia is a crucial event in the infectious process of S. apiospermum, we studied the morphological transformation induced by the incubation of conidia in Sabouraud-dextrose medium at 37 degrees C. After 6 h, some conidia presented a small projection resembling a germ-tube. A significant increase, around sixfold, in the germ-tube length was found after 12 h, and hyphae were exclusively observed after 24 h. Three distinct metallopeptidase inhibitors were able to arrest the transformation of conidia into hyphae in different ways; for instance, 1,10-phenanthroline (PHEN) completely blocked this process at 10 mu mol l-1, while ethylenediamine tetraacetic acid (EDTA) and ethylene glycol-bis (beta-aminoethyl ether; EGTA) only partially inhibited the differentiation at up to 10 mmol l-1. EGTA did not promote any significant reduction in the conidial growth, while PHEN and EDTA, both at 10 mmol l-1, inhibited the proliferation around 100% and 65%, respectively. The secretion of polypeptides into the extracellular environment and the metallopeptidase activity secreted by mycelia were completely inhibited by PHEN. These findings suggest that metallo-type enzymes could be potential targets for future therapeutic interventions against S. apiospermum.

Preferential location of lidocaine and etidocaine in lecithin bilayers as determined by EPR, fluorescence and H-2 NMR

We have examined the effect of the uncharged species of lidocaine (LDC) and etidocaine (EDC) on the acyl chain moiety of egg phosphatidylcholine liposomes. Changes in membrane organization caused by both anesthetics were detected through the use of EPR spin labels (5, 7 and 12 doxyl stearic acid methyl ester) or fluorescence probes (4, 6, 10, 16 pyrene-fatty acids). The disturbance caused by the LA was greater when the probes were inserted in more external positions of the acyl chain and decreased towards the hydrophobic core of the membrane. The results indicate a preferential insertion of LDC at the polar interface of the bilayer and in the first half of the acyl chain, for EDC. Additionally, 2 H NMR spectra of multilamellar liposomes composed by acyl chain-perdeutero DMPC and EPC (1:4 mol%) allowed the determination of the segmental order (S-mol) and dynamics (T-1) of the acyl chain region. In accordance to the fluorescence and EPR results, changes in molecular orientation and dynamics are more prominent if the LA preferential location is more superficial, as for LDC while EDC seems to organize the acyl chain region between carbons 2-8, which is indicative of its positioning. We propose that the preferential location of LDC and EDC inside the bilayers creates a ""transient site"", which is related to the anesthetic potency since it could modulate the access of these molecules to their binding site(s) in the voltage-gated sodium channel. (C) 2007 Elsevier B.V. All rights reserved.

Fungos anemófilos em Porto Alegre, RS.

O conhecimento dos fungos anemófilos em determinada cidade ou região é importante para o diagnóstico etiológico e o tratamento específico das manifestações alérgicas respiratórias. Várias técnicas quantitativas e qualitativas são preconizadas para coleta e identificação desses fungos na dependência do local estudado. Nesta pesquisa, sobre esporos de fungos do ar, foi utilizado o equipamento Rotorod Sampler®, que retira a amostra do ar através de um bastão de plástico preso a um motor elétrico que o faz girar rapidamente, sendo as partículas suspensas no ar recolhidas pelo bastão. As amostras foram coletadas uma vez por semana, durante 24 horas, correspondendo a um ciclo de coleta. Foram realizadas 52 coletas entre abril de 2000 e março de 2001. Os resultados mostraram prevalência de ascosporos (50,49%), Cladosporium (17,86%), Aspergillus/Penicillium (15,03%), basidiosporos (3,84%), rusts (3,82%), Helminthosporium (2,49%), Botrytis (1,22%), Alternaria (1,19%), smuts (0,90%), Curvularia (0,87%), Nigrospora (0,61%) e Fusarium (0,08%). Não foi possível identificar 1,59% dos esporos de fungos coletados no período. O maior número de esporos foi observado nos meses de verão e o menor, durante o outono. Utilizando provas in vivo e in vitro, avaliou-se a hipersensibilidade a fungos entre 39 pacientes atópicos sofrendo de rinite e ou asma brônquica. Os testes cutâneos identificaram sensibilização em 17,94% dos pacientes, enquanto as provas sorológicas caracterizaram presença de IgE específica em 12,82% dos casos avaliados. A detecção de significativo número de esporos de fungos no ar de Porto Alegre, com muitas espécies comprovadamente alergênicas, e os índices de sensibilização observados em indivíduos atópicos confirmam a importância do estudo dos fungos anemófilos nessa cidade, com vistas a aprimorar o diagnóstico e o manejo de pacientes com manifestações alérgicas respiratórias.

Estudo da resistência do streptococcus pneumoniae à penicilina em pneumopatias infecciosas nas cidades de Porto Alegre e Caxias do Sul (RS - Brasil)

A resistência aos antibióticos dos patógenos mais comuns do trato respiratório está aumentando mundialmente. Recentemente, Streptococcus pneumoniae resistente à penicilina tem sido isolado em diversos países, e a freqüência dessas cepas tem elevado de modo alarmante. O aumento da resistência, com conseqüentes implicações terapêuticas, tem levado a uma reavaliação do uso dos antibióticos ß-lactâmicos para o tratamento de infecções pneumocócicas. No presente trabalho, um total de 107 amostras de Streptococcus pneumoniae, obtidas de materiais provenientes de pacientes adultos ambulatoriais e hospitalizados, em dois centros médicos de duas cidades do Rio Grande do Sul (Porto Alegre e Caxias do Sul), os quais apresentavam quadro clínico-radiológico de infecção pulmonar, foram analisadas com o objetivo de estudar-se a resistência do germe à penicilina. As amostras constituídas de escarro (80,4%), lavado brônquico (13,5%) e aspirado traqueal (6,6%) foram coletadas no período compreendido entre Julho de 1998 e Julho de 1999. O material foi semeado em meio de Agar sangue e as colônias suspeitas de Streptococcus pneumoniae foram transferidas para meio de Mueller-Hinton para teste de optoquina e de sensibilidade à penicilina com discos de oxacilina. Um halo de inibição da oxacilina menor do que 20 mm indicava a realização de teste para determinação da concentração inibitória mínima (MIC) com E-test. Um total de nove cepas foi identificado como tendo resistência intermediária à penicilina (MIC 0,1-1,0μg/ml) e nenhuma cepa resistente (resistência elevada: MIC > 2,0 μg/ml) foi identificada. Uma monitorização local das cepas quanto à resistência antimicrobiana é de grande importância para os clínicos no manejo de infecções pneumocócicas.

Preferential location of prilocaine and etidocaine in phospholipid bilayers: A molecular dynamics study

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Preferential location of lidocaine and etidocaine in lecithin bilayers as determined by EPR, fluorescence and H-2 NMR

We have examined the effect of the uncharged species of lidocaine (LDC) and etidocaine (EDC) on the acyl chain moiety of egg phosphatidylcholine liposomes. Changes in membrane organization caused by both anesthetics were detected through the use of EPR spin labels (5, 7 and 12 doxyl stearic acid methyl ester) or fluorescence probes (4, 6, 10, 16 pyrene-fatty acids). The disturbance caused by the LA was greater when the probes were inserted in more external positions of the acyl chain and decreased towards the hydrophobic core of the membrane. The results indicate a preferential insertion of LDC at the polar interface of the bilayer and in the first half of the acyl chain, for EDC. Additionally, 2 H NMR spectra of multilamellar liposomes composed by acyl chain-perdeutero DMPC and EPC (1:4 mol%) allowed the determination of the segmental order (S-mol) and dynamics (T-1) of the acyl chain region. In accordance to the fluorescence and EPR results, changes in molecular orientation and dynamics are more prominent if the LA preferential location is more superficial, as for LDC while EDC seems to organize the acyl chain region between carbons 2-8, which is indicative of its positioning. We propose that the preferential location of LDC and EDC inside the bilayers creates a "transient site", which is related to the anesthetic potency since it could modulate the access of these molecules to their binding site(s) in the voltage-gated sodium channel. (C) 2007 Elsevier B.V. All rights reserved.

Eugenol em juvenis de tilápia do nilo: concentrações e administrações sucessivas

Several anesthetics are used in aquaculture to reduce the stress caused by management. In the field, the most common anesthetic is benzocaine, but several authors are researching about eugenol. The objective of this study were to compare the action of these drugs on the anesthetics stages and measure the effects of successive administration of eugenol on the anesthetics stages in Oreochromis niloticus. In the first test, fish (47.73 +/- 8.73 g and 14.23 +/- 0.81 cm) were subjected to five concentrations of eugenol (40, 60, 80, 100 and 120 mg L-1) and one of benzocaine (100 mg L-1). All the anesthetic concentrations tested were able to induce fish to all stages of anesthesia, and revealed an inverse relationship between the induction times and increased concentrations. The minimum concentration of eugenol to induce anesthesia was 100 mg L-1. In the second test, the juveniles (38.07 +/- 5.00 g and 12.70 +/- 0.54 cm) were exposed to concentrations of 60, 80, 100 mg L-1 of eugenol, as determined in the first test. Evaluating the differences between successive days, there was no significant difference (P > 0.05) between anesthetics stages in the concentrations of eugenol tested, suggesting that the residue of eugenol was removed from the fish in less than 24 hours.

Cardiopulmonary and acid-base effects of desflurane and sevoflurane in spontaneously breathing cats

The cardiopulmonary effects of desflurane and sevoflurane anesthesia were compared in cats breathing spontaneously. Heart (HR) and respiratory (RR) rates; systolic (SAP), diastolic (DAP) and mean arterial (MAP) pressures; partial pressure of end tidal carbon dioxide (PETCO(2)), arterial blood pH (pH), arterial partial pressure of oxygen (PaO(2)) and carbon dioxide (PaCO(2)); base deficit (BD), arterial oxygen saturation (SaO(2)) and bicarbonate ion concentration (HCO(3)) were measured. Anesthesia was induced with propofol (8 +/- 2.3 mg/kg IV) and maintained with desflurane (GD) or sevoflurane (GS), both at 1.3 MAC. Data were analyzed by analysis of variance (ANOVA), followed by the Tukey test (P < 0.05). Both anesthetics showed similar effects. HR and RR decreased when compared to the basal values, but remained constant during inhalant anesthesia and PETCO(2) increased with time. Both anesthetics caused acidemia and hypercapnia, but BD stayed within normal limits. Therefore, despite reducing HR and SAP (GD) when compared to the basal values, desflurane and sevoflurane provide good stability of the cardiovascular parameters during a short period of inhalant anesthesia (T20-T60). However, both volatile anesthetics cause acute respiratory acidosis in cats breathing spontaneously. (c) 2004 ESFM and AAFP. Published by Elsevier Ltd. All rights reserved.

Effects of the Ottonia martiana Miq. (Piperaceae) extract on dog's ocular surface

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effects of isoflurane on echocardiographic parameters in healthy dogs

Obejective To study the echocardiographic effects of isoflurane at an end-tidal concentration approximating 1.0 times the minimum alveolar concentration (MAC) in healthy unpremedicated dogs.Study design Prospective experimental trial.Animals Sixteen mature mongrel dogs of either sex weighing 11.06 +/- 2.72 kg.Methods After performing a baseline echocardiogram in the awake animal, anesthesia was induced with increasing inspired concentrations of isoflurane via a face mask until tracheal intubation was possible. Following intubation, the end-tidal concentration was decreased to 1.4% for the rest of the anesthetic period. Serial echocardiograms were recorded at 25, 40, and 55 minutes after the end-tidal concentration was reached.Results No changes were observed in heart rate. However, significant decreases were seen in left ventricular end-diastolic diameter (Mean maximal change: 13.8%), interventricular septal thickness during systole (15.2%), interventricular septal thickening fraction (72.2%), left ventricular free wall thickening fraction (63.5%), ejection fraction (39.9%), and fractional shortening (46.7%). In addition, peak flow velocities across mitral, pulmonic, and aortic valves were significantly lower than baseline values. Decreases were also observed in end-diastolic left ventricular volume index (approximately 32.1% from the awake value), stroke index (58.2%), and cardiac index (55.3%) when compared with awake measurements.Conclusions Our results indicate that 1 x MAC isoflurane caused significant myocardial depression in healthy dogs. These changes in myocardial function need to be considered carefully when isoflurane is to be used in dogs with poor cardiovascular reserve.

Comportamento da pressão intra-ocular segundo os efeitos cardiorrespiratórios e hemodinâmicos induzidos pela anestesia com desflurano, em cães submetidos à hipovolemia experimental

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Efeitos do desfluorano, sevofluorano e isofluorano sobre variáveis respiratórias e hemogasométricas em cães

Com este estudo objetivou-se avaliar, comparativamente, os efeitos de três anestésicos inalatórios sobre variáveis respiratórias e hemogasométricas em cães. Para tal, utilizaram-se 30 cães sadios, adultos, machos e fêmeas evitando-se aquelas em estro ou em gestação. Os animais foram separados e protocolados em três grupos de 10 cães cada (G1, G2 e G3). Induziu-se a anestesia geral com administração intravenosa de propofol, na dose de 10±1,3 mg/kg. em seguida procedeu-se à intubação orotraqueal e anestesia geral inalatória pelo desfluorano (G1), sevofluorano (G2) e isofluorano (G3), diluídos em oxigênio a 100,00%, por meio de circuito anestésico tipo semi-fechado, dotado de vaporizadores calibrados para cada agente anestésico. As variáveis estudadas foram Freqüência Respiratória, CO2 ao Final da Expiração, Saturação de Oxihemoglobina, Volume Corrente, Volume Minuto, Pressão Parcial Arterial de O2, Pressão Parcial Arterial de CO2, Excesso de Bases e pH. A avaliação estatística destas variáveis foi realizada pela Análise de Perfil, sendo considerado o nível de significância de 5,00%. Os resultados obtidos permitiram concluir que o desfluorano deprime o sistema respiratório aumentando a pressão parcial arterial de CO2 e o CO2 ao final da expiração; e diminuindo a pressão parcial arterial de O2 e o volume minuto, quando comparado com os outros anestésicos em teste.

Treinamento muscular inspiratório para asma: revisão sistemática com metanálise

In asthmatic, the lung hyperinflation leaves the inspiratory muscle at a suboptimal position in length-tension relationship, reducing the capacity of to generate tension. The increase in transversal section area of the inspiratory muscles could reverse or delay the deterioration of inspiratory muscle function. Objective: To evaluate the evidence for the efficacy of inspiratory muscle training (IMT) with an external resistive device in patients with asthma. Methods: A systematic review with meta-analysis was carried out. The sources researched were the Cochrane Airways Group Specialised Register of trials, Cochrane Central Register of Controlled Trials (The Cochrane Library Issue 11 of 12, 2012), MEDLINE, EMBASE, PsycINFO, CINAHL, AMED, ClinicalTrials.gov and reference lists of articles. All databases were searched from their inception up to November 2012 and there was no restriction on the language of publication. Randomised controlled trials that involved the use of an external inspiratory muscle training device versus a control (sham or no inspiratory training device) were considered for inclusion. Two reviewers independently selected articles for inclusion, evaluated risk of bias in studies and extracted data. Results: A total of five studies involving 113 asthmatic patients were included. Three clinical trials were produced by the same group. The included studies showed a significant increase in maximal inspiratory pressure (MD 13.34 cmH2O, 95% CI 4.70 to 21.98), although the confidence intervals were wide. There was no statistically significant difference between the IMT group and the control group for maximal expiratory pressure, peak expiratory flow rate, forced expiratory volume in one second, forced vital capacity, sensation of dyspnea and use of beta2-agonist. There were no studies describing exacerbation events that required a course of oral and inhaled corticosteroids or emergency department visits, inspiratory muscle endurance, hospital admissions and days of work or school. Conclusions: There is no conclusive evidence in this review to support or refute inspiratory muscle training for asthma, once the evidence was limited by the small number of studies included, number of participants in them together with the risk of bias. More well conducted randomized controlled trials are needed, such trials should investigate respiratory muscle strength, exacerbation rate, lung function, symptoms, hospital admissions, use of medications and days off work or school. IMT should also be assessed in the context of more severe asthma

Encoding mechanisms based on fast oscillations in the retina of the cat and their dependencies on anesthesia

Processing in the visual system starts in the retina. Its complex network of cells with different properties enables for parallel encoding and transmission of visual information to the lateral geniculate nucleus (LGN) and to the cortex. In the retina, it has been shown that responses are often accompanied by fast synchronous oscillations (30 - 90 Hz) in a stimulus-dependent manner. Studies in the frog, rabbit, cat and monkey, have shown strong oscillatory responses to large stimuli which probably encode global stimulus properties, such as size and continuity (Neuenschwander and Singer, 1996; Ishikane et al., 2005). Moreover, simultaneous recordings from different levels in the visual system have demonstrated that the oscillatory patterning of retinal ganglion cell responses are transmitted to the cortex via the LGN (Castelo-Branco et al., 1998). Overall these results suggest that feedforward synchronous oscillations contribute to visual encoding. In the present study on the LGN of the anesthetized cat, we further investigate the role of retinal oscillations in visual processing by applying complex stimuli, such as natural visual scenes, light spots of varying size and contrast, and flickering checkerboards. This is a necessary step for understanding encoding mechanisms in more naturalistic conditions, as currently most data on retinal oscillations have been limited to simple, flashed and stationary stimuli. Correlation analysis of spiking responses confirmed previous results showing that oscillatory responses in the retina (observed here from the LGN responses) largely depend on the size and stationarity of the stimulus. For natural scenes (gray-level and binary movies) oscillations appeared only for brief moments probably when receptive fields were dominated by large continuous, flat-contrast surfaces. Moreover, oscillatory responses to a circle stimulus could be broken with an annular mask indicating that synchronization arises from relatively local interactions among populations of activated cells in the retina. A surprising finding in this study was that retinal oscillations are highly dependent on halothane anesthesia levels. In the absence of halothane, oscillatory activity vanished independent of the characteristics of the stimuli. The same results were obtained for isoflurane, which has similar pharmacological properties. These new and unexpected findings question whether feedfoward oscillations in the early visual system are simply due to an imbalance between excitation and inhibition in the retinal networks generated by the halogenated anesthetics. Further studies in awake behaving animals are necessary to extend these conclusions

Comparison between S-ketamine and clonidine in caudal analgesia

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)