Dendrimer nanocarriers for transport modulation across models of the pulmonary epithelium

The purpose of this study was to determine the effect of PEGylation on the interaction of poly(amidoamine) (PAMAM) dendrimer nanocarriers (DNCs) with in vitro and in vivo models of the pulmonary epithelium. Generation-3 PAMAM dendrimers with varying surface densities of PEG 1000 Da were synthesized and characterized. The results revealed that the apical to basolateral transport of DNCs across polarized Calu-3 monolayers increases with an increase in PEG surface density. DNC having the greatest number of PEG groups (n = 25) on their surface traversed at a rate 10-fold greater than its non-PEGylated counterpart, in spite of their larger size. This behavior was attributed to a significant reduction in charge density upon PEGylation. We also observed that PEGylation can be used to modulate cellular internalization. The total uptake of PEG-free DNC into polarized Calu-3 monolayers was 12% (w/w) vs 2% (w/w) for that with 25 PEGs. Polarization is also shown to be of great relevance in studying this in vitro model of the lung epithelium. The rate of absorption of DNCs administered to mice lungs increased dramatically when conjugated with 25 PEG groups, thus supporting the in vitro results. The exposure obtained for the DNC with 25PEG was determined to be very high, with peak plasma concentrations reaching 5 mu gmL(-1) within 3 h. The combined in vitro and in vivo results shown here demonstrate that PEGylation can be potentially used to modulate the internalization and transport of DNCs across the pulmonary epithelium. Modified dendrimers thereby may serve as a valuable platform that can be tailored to target the lung tissue for treating local diseases, or the circulation, using the lung as pathway to the bloodstream, for systemic delivery.

A field study to describe diel, tidal and semilunar rhythms of larval release in an assemblage of tropical rocky shore crabs

Available information on the larval release rhythms of brachyurans is biased to temperate estuarine species and outcomes resulting from some sort of artificial manipulation of ovigerous females. In this study we applied field methods to describe the larval release rhythms of an assemblage of tropical rocky shore crabs. Sampling the broods of ovigerous females of Pachygrapsus transversus at two different shores indicated a spatially consistent semilunar pattern, with larval release maxima around the full and new moon. Yet, synchronism between populations varied considerably, with the pattern obtained at the site exposed to a lower wave action far more apparent. Breeding cohorts at one of the sampled shores apparently belonged to actual age groups composing the ovigerous population. The data suggest that these breeding groups release their larvae in alternate syzygy periods, responding to a lunar cycle instead of the semilunar pattern observed for the whole population. For the description of shorter-term rhythms, temporal series at hour intervals were obtained by sampling the plankton and confinement boxes where ovigerous females were held. Unexpectedly, diurnal release activity prevailed over nocturnal hatching. Yet, only grapsids living higher on the shore exhibited strong preferences over the diel cycle, with P. transversus releasing their larvae during the day and Geograpsus lividus during the night. The pea crab Dissodactylus crinitichelis, the spider crab Epialtus brasiliensis and a suite of xanthoids undertook considerable releasing activity in both periods. Apart from the commensal pea crab D. crinitichelis, all other taxa revealed tide-related rhythms of larval release, with average estimates of the time of maximum hatching always around the time of high tides; usually during the flooding and slack, rather than the ebbing tide. Data obtained for P. transversus females held in confinement boxes indicated that early larval release is mostly due to nocturnal hatching, while zoeal release in diurnal groups took place at the time of high tide. Since nocturnal high tides at the study area occurred late, sometimes close to dusk, early release would allow more time for offshore transport of larvae when the action of potential predators is reduced.

Renal and vascular effects of the natriuretic peptide isolated from Crotalus durissus cascavella venom

Crotalus durissus cascavella is a snake that is usually found in the scrublands of northeast Brazil. The components of its venom may have effects on the vascular and renal systems. Recently, a new bradykinin inhibitory peptide has been identified in the venom of the Crotalinae family. The aim of the present study was to investigate the renal and vascular effects of the natriuretic peptide isolated from the venom of Crotalus durissus cascavella (NP2_Casca). The chromatographic profile showed the fractionation of substances identified as convulxin, gyroxin, crotoxin and crotamine, as well as fractions V and VI. The electrophoretic profile of fraction V consisted of several bands ranging from approximately 6 kDa to 13 kDa, while fraction VI showed only two main electrophoretic bands with molecular weights of approximately 6 and 14 kDa. Reverse-phase chromatography showed that NP2_Casca corresponds to about 18% of fraction VI and that this fraction is the main natriuretic peptide. NP2_Casca was compared to other natriuretic peptides from other sources of snake venom. All amino acid sequences that were compared showed a consensus region of XGCFGX, XLDRIX and XSGLGCX. The group treated with NP2-Casca showed an increase in perfusion pressure, renal vascular resistance, urinary flow and glomerular filtration rate. The percent of total and proximal tubular transport of sodium was reduced significantly after administration of the peptide. The mean arterial pressure showed a dose-dependent decrease after infusion of NP2_Casca, and an increase in nitrite production. In the aortic ring assay, NP2_Casca caused a relaxant effect in endothelium-intact thoracic aortic rings precontracted with phenylephrine in the presence and absence of isatin. NP2_Casca failed to relax the aortic rings precontracted with an isosmotic potassium Krebs-Henseleit solution. In conclusion, the natriuretic peptide isolated from Crotalus durissus cascavella venom produced renal and vascular effects. NP2_Casca reduced total and proximal sodium tubular transport, leading to an increase in sodium excretion, thereby demonstrating a diuretic action. A hypotensive effect was displayed in an arterial pressure assay, with an increase in nitrite production, suggesting a possible vasoactive action. (C) 2008 Elsevier Ltd. All rights reserved.

Transport stress in matrinxa, Brycon cephalus (Teleostei : Characidae), at different densities

Matrinxa, Brycon cephalus, is a native teleost fish from the Amazon Basin, and is of economic importance for cultivation for food and sport in Brazil. Mortality losses due to handling and transport of this stenohaline freshwater species are common. The effects of transportation at different densities on the biochemical stress responses of matrinxa (mean weight 1 kg) were examined. Fish were subjected to three different transport densities (100, 200, and 300 kg m(-3)) for four h in water with added salt (0.6%). The fish were bled at departure (baseline level), arrival (immediately after transportation) and at 24 and 96 h after arrival (recovery period). Blood glucose, cortisol, sodium, chloride, potassium and ammonia were used as stress bioindicators. No mortality was observed and no alterations in plasma cortisol were registered. However, blood glucose and ammonia levels increased and serum sodium and plasma chloride decreased on arrival for the fish transported at the highest densities. These stress responses were transient and the concentrations returned to baseline levels within 24 h. This study showed that matrinxa can be transported at densities as high as those tested in the present study, at least under the conditions employed in this study. A recovery period of at least 24 h is strongly recommended.

Spatial variability of soil CO2 emission in different topographic positions

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

Viabilidade econômica dos sistemas manual e mecanizado de aplicação de herbicidas em ferrovias

A aplicação mecanizada de herbicidas em ferrovias tem como principal vantagem a grande capacidade operacional, cobrindo trechos extensos em curtos espaços de tempo. Entretanto em função das características dos equipamentos utilizados, quase sempre ocorre grande desperdício de herbicida. A aplicação manual, através de pulverizadores costais, tem como principal vantagem a possibilidade da pulverização dirigida às áreas infestadas, diminuindo a quantidade de herbicida aplicado. Por outro lado, sua reduzida capacidade operacional faz com que a aplicação de longos trechos seja demorada, tornando-a, assim, mais dependente dos problemas climáticos e operacionais da ferrovia. O objetivo do trabalho foi realizar uma análise comparativa das características operacionais e econômicas destes sistemas de aplicação, apontando os fatores mais importantes a serem avaliados para a sua implantação. Para tanto, foram realizadas simulações em computador, baseando-se em diversas equações matemáticas e dados levantados no campo. Os resultados mostraram que a aplicação manual apresentou-se mais econômica apenas para trechos com baixas infestações (até 10%, e média). Na composição de custos, os gastos com herbicidas corresponderam a cerca de 80% do total para a aplicação mecanizada e apenas 8% na manual. Levando-se em conta os fatores operacionais e de mão-de-obra, em apenas um dos trechos avaliados houve vantagem econômica do uso da aplicação manual.

Internal Architecture and Population Size of Acromyrmex subterraneus molestans (Hymenoptera, Formicidae) Nests: Comparison Between a Rural and an Urban Area

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Observation of persistent photoconductivity in vanadyl phthalocyanine

Persistent photoconductivity (PPC) in vanadyl phthalocyanine (VOPc) organic light-emitting diodes was investigated using photoconductive time response, photocurrent-voltage characteristics and charge extraction in linearly increasing voltage (CELIV) measurements. The experiments were performed in phase 1 (amorphous) and in phase 2 (crystalline) samples obtained by the physical vapour deposition (PVD) technique over ITO/glass electrodes with an Al covering electrode. The results indicated a photoconductivity with a long decay time in phase 1 VOPc described by a stretched exponential relaxation. The device showed a rectifying behaviour and the mobility of holes was measured by CELIV, following a dispersive model. In crystalline samples the PPC effect was not observed and the dominant mechanism of transport of holes was hopping in a Gaussian density of states.

Effects of buprenorphine, carprofen and saline on thermal and mechanical nociceptive thresholds in cats

To evaluate a prototype pressure stimulus device for use in the cat and to compare with a known thermal threshold device.Eight healthy adult cats weighing between 3.0 and 4.9 kg.Pressure stimulation was given via a plastic bracelet taped around the forearm. Three 2.4 mm diameter ball bearings, in a 10-mm triangle, were advanced against the craniolateral surface of the antebrachium by manual inflation of a modified blood pressure bladder. Pressure in the cuff was recorded at the end point (leg shake and head turn). Thermal threshold was also tested. Stimuli were stopped if they reached 55 degrees C or 450 mmHg without response. After four pressure and thermal threshold baselines, each cat received SC buprenorphine 0.01 mg kg(-1), carprofen 4 mg kg(-1) or saline 0.3 mL in a three period cross-over study with a 1-week interval. The investigator was blinded to the treatment. Measurements were made at 0.25. 0.5, 0.75, 1, 2, 3, 4, 6, 8, and 24 hours after injection. Data were analyzed by using ANOVA.There were no significant changes in thermal or pressure threshold after administration of saline or carprofen, but thermal threshold increased from 60 minutes until 8 hours after administration of buprenorphine (p < 0.05). The maximum increase in threshold from baseline (Delta T-max) was 3.5 +/- 3.1 degrees C at 2 hours. Pressure threshold increased 2 hours after administration of buprenorphine (p < 0.05) when the increase in threshold above baseline (Delta P-max) was 162 +/- 189 mmHg.This pressure device resulted in thresholds that were affected by analgesic treatment in a similar manner but to a lesser degree than the thermal method. Pressure stimulation may be a useful additional method for analgesic studies in cats.

Effects of meperidine or saline on thermal, mechanical and electrical nociceptive thresholds in cats

Objective To measure cutaneous electrical nociceptive thresholds in relation to known thermal and mechanical stimulation for nociceptive threshold detection in cats.Study design Prospective, blinded, randomized cross-over study with 1-week washout interval.Animals Eight adult cats [bodyweight 5.1 +/- 1.8 kg (mean + SD)].Methods Mechanical nociceptive thresholds were tested using a step-wise manual inflation of a modified blood pressure bladder attached to the cat's thoracic limb. Thermal nociceptive thresholds were measured by increasing the temperature of a probe placed on the thorax. The electrical nociceptive threshold was tested using an escalating current from a constant current generator passed between electrodes placed on the thoracic region. A positive response (threshold) was recorded when cats displayed any or all of the following behaviors: leg shake, head turn, avoidance, or vocalization. Four baseline readings were performed before intramuscular injection of meperidine (5 mg kg(-1)) or an equal volume of saline. Threshold recordings with each modality were made at 15, 30, 45, 60, 90, and 120 minutes post-injection. Data were analyzed using ANOVA and paired t-tests (significance at p < 0.05).Results There were no significant changes in thermal, mechanical, or electrical thresholds after saline. Thermal thresholds increased at 15-60 minutes (p < 0.01) and mechanical threshold increased at 30 and 45 minutes after meperidine (p < 0.05). Maximum thermal threshold was +4.1 +/- 0.3 degrees C above baseline at 15 minutes while maximum mechanical threshold was 296 +/- 265 mmHg above baseline at 30 minutes after meperidine. Electrical thresholds following meperidine were not significantly different than baseline (p > 0.05). Thermal and electrical thresholds after meperidine were significantly higher than saline at 30 and 45 minutes (p < 0.05), and at 120 minutes (p < 0.05), respectively. Mechanical thresholds were significantly higher than saline treatment at 30 minutes (p <= 0.05).Conclusion and clinical relevance Electrical stimulation did not detect meperidine analgesia whereas both thermal and mechanical thresholds changed after meperidine administration in cats.

Molecular detection of Streptococcus agalactiae in bovine raw milk samples obtained directly from bulk tanks

Mastitis is the most common infectious disease affecting dairy cattle; in addition, it remains the most economically important disease of dairy industries around the world. Streptococcus agalactiae, a contagious pathogen associated with subclinical mastitis, is highly infectious. This bacterium can cause an increase in bulk tank bacterial counts (BTBC) and bulk tank somatic cell counts (BTSCC). The microbiological identification of S. agalactiae in samples from bulk tanks is an auxiliary method to control contagious mastitis. Thus, there are some limitations for time-consuming cultures or identification methods and additional concerns about the conservation and transport of samples. Bulk tank samples from 247 dairy farms were cultured and compared through polymerase chain reaction (PCR), directed to 16S rRNA genes of S. agalactiae, followed by BTBC and S. agalactiae isolation. The mean value of BTBC was 1.08 x 10(6) CFU mL(-1) and the bacterium was identified through the microbiological method in 98 (39.7%; CI95% = 33.8-45.9%) and through PCR in 110 (44.5%; CI95% = 38.5-50.8%) samples. Results indicated sensitivity of 0.8571 +/- 0.0353 (CI95% = 0.7719-0.9196) and specificity of 0.8255 +/- 0.0311 (CI95% = 0.7549-0.8827). The lack of significant difference between microbiological and molecular results (kappa = 0.6686 +/- 0.0477 and CI95% = 0.5752-0.7620) indicated substantial agreement between the methods. This suggests that PCR can be used for bulk tank samples to detect contagious mastitis caused by S. agalactiae. (C) 2011 Elsevier Ltd. All rights reserved.

Storage sites in seeds of Caesalpinia echinata and C. ferrea (Leguminosae) with considerations on nutrients flow

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

Allochthonous matter input in two different stretches of a headstream (Itatinga, São Paulo, Brazil)

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

Input of litter in deforested and forested areas of a tropical headstream

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Ultrastructural study of the follicular epithelium in oocytes of Pachycondyla (Neoponera) villosa ants (Hymenoptera : Ponerinae)

Insect oocytes are surrounded by the follicular epithelium which is simple and cuboidal, wih the mainly functions of: synthesis of vitellin membrane and chorion and synthesis and transport of hemolymph products (proteins). In Pachycondyla (Neoponera) villosa ants workers aged less than 10 days do not present the formation of ovarian follicles (oocytes, nurse cells and follicular cells) indicating that vitellogenesis starts at approximately 10 days of age. Studies of participation of the follicular epithelium in Pachycondyla (Neoponera) villosa showed that in stage I oocytes the epithelium does not present the opening of intercellular spaces. In stage II these spaces begin to be observed together with separation of the follicular epithelium from the oocyte surface. In stage III two types of material were observed in the intercellular spaces: electrodense material in the basal region and compacted material in apical one as well as follicular epithelium/oocytes interface suggesting that the extraovarian material that reach oocytes undergoes some type of modification during passage through the intercellular spaces. The follicular epithelium spaces in queen are bigger than in workers oocytes.