Growth regulators and their reflection on different hop genotypes cultivated under in vitro conditions / Reguladores de crescimento e seu reflexo em diferentes genótipos de lúpulo cultivados in vitro

Hops is a new culture in Brazil. Tissue culture can be an important technique for rapid hop propagation. This paper aims to characterize responses from different genotypes under different growth regulators through the interrelationship of response variables important to hop in vitro growth. Three genotypes were cultivated in six culture media with different combinations of growth regulators, BAP (6-benzylaminopurine), IAA (3-indolacetic acid) and GA3 (gibberellic acid). The means were compared by orthogonal contrasts and the interrelationship of the response variables was performed by path analysis. American genotypes showed favorable root development under the BAP + IAA combination, while the use of IAA improved shoot development. The origin of genotypes was important for defining the best protocol for in vitro cultivation. The path coefficient showed that the variable number of shoots has stronger direct effect on the number of nodal segments. Additionally, in tissue culture assays, the use of a covariable and proper error distribution significantly increased experimental accuracy.

O lúpulo é uma nova cultura no Brasil. A cultura de tecidos pode ser uma técnica importante para a propagação rápida do lúpulo. Este artigo tem como objetivo caracterizar respostas de diferentes genótipos sob diferentes reguladores de crescimento através da inter-relação de variáveis de resposta importantes para o crescimento in vitro. Três genótipos foram cultivados em seis meios de cultura com diferentes combinações de reguladores de crescimento, BAP (6-benzilaminopurina), AIA (ácido 3-indolacético) e GA3 (ácido giberélico). As médias foram comparadas por contrastes ortogonais e a inter-relação das variáveis de resposta foi realizada por análise de trilha. Os genótipos americanos apresentaram desenvolvimento radicular favorável sob a combinação BAP + AIA, enquanto o uso do AIA melhorou o desenvolvimento da parte aérea. A origem dos genótipos foi importante para definir o melhor protocolo para o cultivo in vitro. O coeficiente de trilha mostrou que a variável número de brotos tem um efeito direto mais forte no número de segmentos nodais. Adicionalmente, em experimentos com cultura de tecidos, o uso de uma covariável e distribuição de erro adequada aumentou significativamente a precisão experimental.

Growth regulators and their reflection on different hop genotypes cultivated under in vitro conditions

Abstract Hops is a new culture in Brazil. Tissue culture can be an important technique for rapid hop propagation. This paper aims to characterize responses from different genotypes under different growth regulators through the interrelationship of response variables important to hop in vitro growth. Three genotypes were cultivated in six culture media with different combinations of growth regulators, BAP (6-benzylaminopurine), IAA (3-indolacetic acid) and GA3 (gibberellic acid). The means were compared by orthogonal contrasts and the interrelationship of the response variables was performed by path analysis. American genotypes showed favorable root development under the BAP + IAA combination, while the use of IAA improved shoot development. The origin of genotypes was important for defining the best protocol for in vitro cultivation. The path coefficient showed that the variable number of shoots has stronger direct effect on the number of nodal segments. Additionally, in tissue culture assays, the use of a covariable and proper error distribution significantly increased experimental accuracy.

Resumo O lúpulo é uma nova cultura no Brasil. A cultura de tecidos pode ser uma técnica importante para a propagação rápida do lúpulo. Este artigo tem como objetivo caracterizar respostas de diferentes genótipos sob diferentes reguladores de crescimento através da inter-relação de variáveis de resposta importantes para o crescimento in vitro. Três genótipos foram cultivados em seis meios de cultura com diferentes combinações de reguladores de crescimento, BAP (6-benzilaminopurina), AIA (ácido 3-indolacético) e GA3 (ácido giberélico). As médias foram comparadas por contrastes ortogonais e a inter-relação das variáveis de resposta foi realizada por análise de trilha. Os genótipos americanos apresentaram desenvolvimento radicular favorável sob a combinação BAP + AIA, enquanto o uso do AIA melhorou o desenvolvimento da parte aérea. A origem dos genótipos foi importante para definir o melhor protocolo para o cultivo in vitro. O coeficiente de trilha mostrou que a variável número de brotos tem um efeito direto mais forte no número de segmentos nodais. Adicionalmente, em experimentos com cultura de tecidos, o uso de uma covariável e distribuição de erro adequada aumentou significativamente a precisão experimental.

Growth regulators and their reflection on different hop genotypes cultivated under in vitro conditions / Reguladores de crescimento e seu reflexo em diferentes genótipos de lúpulo cultivados in vitro

Hops is a new culture in Brazil. Tissue culture can be an important technique for rapid hop propagation. This paper aims to characterize responses from different genotypes under different growth regulators through the interrelationship of response variables important to hop in vitro growth. Three genotypes were cultivated in six culture media with different combinations of growth regulators, BAP (6-benzylaminopurine), IAA (3-indolacetic acid) and GA3 (gibberellic acid). The means were compared by orthogonal contrasts and the interrelationship of the response variables was performed by path analysis. American genotypes showed favorable root development under the BAP + IAA combination, while the use of IAA improved shoot development. The origin of genotypes was important for defining the best protocol for in vitro cultivation. The path coefficient showed that the variable number of shoots has stronger direct effect on the number of nodal segments. Additionally, in tissue culture assays, the use of a covariable and proper error distribution significantly increased experimental accuracy.

O lúpulo é uma nova cultura no Brasil. A cultura de tecidos pode ser uma técnica importante para a propagação rápida do lúpulo. Este artigo tem como objetivo caracterizar respostas de diferentes genótipos sob diferentes reguladores de crescimento através da inter-relação de variáveis de resposta importantes para o crescimento in vitro. Três genótipos foram cultivados em seis meios de cultura com diferentes combinações de reguladores de crescimento, BAP (6-benzilaminopurina), AIA (ácido 3-indolacético) e GA3 (ácido giberélico). As médias foram comparadas por contrastes ortogonais e a inter-relação das variáveis de resposta foi realizada por análise de trilha. Os genótipos americanos apresentaram desenvolvimento radicular favorável sob a combinação BAP + AIA, enquanto o uso do AIA melhorou o desenvolvimento da parte aérea. A origem dos genótipos foi importante para definir o melhor protocolo para o cultivo in vitro. O coeficiente de trilha mostrou que a variável número de brotos tem um efeito direto mais forte no número de segmentos nodais. Adicionalmente, em experimentos com cultura de tecidos, o uso de uma covariável e distribuição de erro adequada aumentou significativamente a precisão experimental.

Growth regulators and their reflection on different hop genotypes cultivated under in vitro conditions.

Hops is a new culture in Brazil. Tissue culture can be an important technique for rapid hop propagation. This paper aims to characterize responses from different genotypes under different growth regulators through the interrelationship of response variables important to hop in vitro growth. Three genotypes were cultivated in six culture media with different combinations of growth regulators, BAP (6-benzylaminopurine), IAA (3-indolacetic acid) and GA3 (gibberellic acid). The means were compared by orthogonal contrasts and the interrelationship of the response variables was performed by path analysis. American genotypes showed favorable root development under the BAP + IAA combination, while the use of IAA improved shoot development. The origin of genotypes was important for defining the best protocol for in vitro cultivation. The path coefficient showed that the variable number of shoots has stronger direct effect on the number of nodal segments. Additionally, in tissue culture assays, the use of a covariable and proper error distribution significantly increased experimental accuracy.

The last frontier: Coupling technological developments with scientific challenges to improve hazard assessment of deep-sea mining.

The growing economic interest in the exploitation of mineral resources on deep-ocean beds, including those in the vicinity of sensitive-rich habitats such as hydrothermal vents, raise a mounting concern about the damage that such actions might originate to these poorly-know ecosystems, which represent millions of years of evolution and adaptations to extreme environmental conditions. It has been suggested that mining may cause a major impact on vent ecosystems and other deep-sea areas. Yet, the scale and the nature of such impacts are unknown at present. Hence, building upon currently available scientific information it is crucial to develop new cost-effective technologies embedded into rigorous operating frameworks. The forward-thinking provided here will assist in the development of new technologies and tools to address the major challenges associated with deep sea-mining; technologies for in situ and ex situ observation and data acquisition, biogeochemical processes, hazard assessment of deep-sea mining to marine organisms and development of modeling tools in support of risk assessment scenarios. These technological developments are vital to validate a responsible and sustainable exploitation of the deep-sea mineral resources, based on the precautionary principle.

A comparative study of electronic stethoscopes for cardiac auscultation.

There are several electronic stethoscopes available on the market today, with a very high potential for healthcare namely telemedicine, assisted decision and education. However, there are no recent comparatives studies published about the recording quality of auscultation sounds. In this study we aim to: a) define a ranking, according to experts opinion of 6 of the most relevant electronic stethoscopes on the market today; b) verify if there are any relations between a stethoscope's performance and the type of pathology present; c) analyze if some pathologies are more easily identified than others when using electronic auscultation. Our methodology consisted in creating two study groups: the first group included 18 cardiologists and cardiology house officers, acting as the gold standard of this work. The second included 30 medical students. Using a database of heart sounds recorded in real hospital environments, we applied questionnaires to observers from each group. The first group listened to 60 cardiac auscultations recorded by the 6 stethoscopes, and each one was asked to identify the pathological sound present: aortic stenosis, mitral regurgitation or normal. The second group was asked to choose, between two auscultation recordings, using as criteria the best sound quality for the identification of pathological sounds. Results include a total of 1080 evaluations, in which 72% of cases were correctly diagnosed. A detailed breakdown of these results is presented in this paper. As conclusions, results showed that the impact of the differences between stethoscopes is very small, given that we did not find statistically significant differences between all pairs of stethoscopes. Normal sounds showed to be easier to identify than pathological sounds, but we did not find differences between stethoscopes in this identification.

Safety and outcome of rtPA in acute ischemic stroke in patients with active cancer: case-control study. / Seguridad y efectividad del rtPA en el ictus isquemico agudo en pacientes con cancer activo: estudio de casos y controles.

INTRODUCTION: Cancer patients have increased stroke risk from direct and indirect malignancy effects. Intravenous thrombolysis with recombinant tissue plasminogen activator (rtPA) is standard medical treatment for acute ischemic stroke (AIS). AIM: To review rtPA use in AIS patients with active cancer. SUBJECTS AND METHODS: Retrospective observational case-control study evaluating patients with AIS and cancer admitted to our stroke unit between January/2010 and June/2015. RESULTS: Seven cases were identified (86% male; median age: 76), and 20 controls were included matched for age and Oxfordshire Community Stroke Project classification. 29% experienced direct procedure complications vs 30% within the control group, 14% suffered haemorrhagic transformation (vs 20%), one patient experienced serious systemic haemorrhage (case) and one patient experienced serious intracerebral haemorrhage (control). After three months' follow-up, 43% were independent compared with 25% controls, and 29% had died (vs 30%). Undetermined aetiology subtype (TOAST classification) was more frequent in cancer patients when compared to controls (71% vs 20%). CONCLUSION: Severe haemorrhagic complications, potentiated by rtPA, carry increased risk of morbidity and mortality. Nevertheless, selected cancer patients with AIS may benefit from rtPA treatment. Active cancer should not be considered an absolute contraindication to rtPA use. Risk of complications and life expectancy should be assessed when making this decision.

TITLE: Seguridad y efectividad del rtPA en el ictus isquemico agudo en pacientes con cancer activo: estudio de casos y controles.Introduccion. Los pacientes con cancer tiene un mayor riesgo de ictus debido a los efectos malignos directos e indirectos. La trombolisis intravenosa con activador tisular del plasminogeno recombinante (rtPA) constituye un tratamiento medico estandar para el ictus isquemico agudo. Objetivo. Revisar el uso de rtPA en el ictus isquemico agudo en pacientes con cancer activo. Sujetos y metodos. Estudio retrospectivo observacional de casos y controles para evaluar pacientes con ictus isquemico agudo y cancer admitidos en la unidad de ictus entre enero de 2010 y junio de 2015. Resultados. Se identificaron siete casos (86% varones; mediana de edad: 76 años) y tambien se incluyeron 20 controles pareados por edad y clasificacion del Oxfordshire Community Stroke Project. Un 29% de casos experimento complicaciones directas del procedimiento frente a un 30% en el grupo control. Un 14% sufrio transformacion hemorragica (frente a un 20%). Un paciente (caso) sufrio una hemorragia sistemica grave, y otro (control), una hemorragia intracerebral grave. A los tres meses, un 43% era independiente (frente a un 25% de los controles) y un 29% habia fallecido (frente a un 30%). Un subtipo etiologico indeterminado (clasificacion TOAST) era mas frecuente en pacientes con cancer (71% frente a 20%). Conclusion. Complicaciones hemorragicas graves, potenciadas por el rtPA, pueden incrementar el riesgo de morbilidad y mortalidad. Sin embargo, pacientes seleccionados con cancer que padecen un ictus isquemico agudo pueden beneficiarse del tratamiento con rtPA. Un cancer activo no deberia considerarse una contraindicacion de uso de rtPA, aunque debe evaluarse el riesgo de complicaciones y la esperanza de vida para tomar la decision.

Cortisol plays a role in the high environmental ammonia associated suppression of the immune response in zebrafish.

Exposure to high environmental ammonia (HEA) levels increases the vulnerability of fishes to parasitic, viral and bacterial diseases. We tested the hypothesis that elevated plasma cortisol levels play a role in the HEA-mediated immunosuppression in fishes. To this end, we tested the effect of exogenous cortisol treatment on the lipopolysaccharide (LPS)-induced immune response in zebrafish (Danio rerio). Also, to test whether glucocorticoid receptor (GR) signaling is involved in HEA-mediated immunosuppression, zebrafish were treated with mifepristone, a GR antagonist, and the LPS-induced immune response assessed after HEA exposure. We evaluated a panel of important immunity-related genes including interleukin 1ß (il1b) and suppressor of cytokine signaling (socs-1a, 2, 3) and acute phase response genes [serum amyloid A (saa), transferrin (tfa), leukocyte cell-derived chemotaxin 2-like (lect2l), haptoglobin (hp), hepcidin (=hepatic anti-microbial peptide hamp), and complement component 3b (c3b)] by real-time quantitative PCR. Our results demonstrate that exogenous cortisol administration as well as elevated cortisol levels in response to HEA exposure modulate mRNA transcript levels of key mediators of the innate immune response in zebrafish. Mifepristone treatment reduced whole body cortisol levels and eliminated the HEA-mediated changes in transcript abundance of socs1a, il1b, as well as APR genes. Together, these results suggest that the HEA effect on the innate immune response is in part mediated by cortisol signaling, while the mode of action, including the receptors involved remains to be elucidated.

A cytosolic carbonic anhydrase molecular switch occurs in the gills of metamorphic sea lamprey.

Carbonic anhydrase plays a key role in CO2 transport, acid-base and ion regulation and metabolic processes in vertebrates. While several carbonic anhydrase isoforms have been identified in numerous vertebrate species, basal lineages such as the cyclostomes have remained largely unexamined. Here we investigate the repertoire of cytoplasmic carbonic anhydrases in the sea lamprey (Petromyzon marinus), that has a complex life history marked by a dramatic metamorphosis from a benthic filter-feeding ammocoete larvae into a parasitic juvenile which migrates from freshwater to seawater. We have identified a novel carbonic anhydrase gene (ca19) beyond the single carbonic anhydrase gene (ca18) that was known previously. Phylogenetic analysis and synteny studies suggest that both carbonic anhydrase genes form one or two independent gene lineages and are most likely duplicates retained uniquely in cyclostomes. Quantitative PCR of ca19 and ca18 and protein expression in gill across metamorphosis show that the ca19 levels are highest in ammocoetes and decrease during metamorphosis while ca18 shows the opposite pattern with the highest levels in post-metamorphic juveniles. We propose that a unique molecular switch occurs during lamprey metamorphosis resulting in distinct gill carbonic anhydrases reflecting the contrasting life modes and habitats of these life-history stages.

Effects of salinity on upstream-migrating, spawning sea lamprey, Petromyzon marinus.

The sea lamprey, Petromyzon marinus, is an anadromous, semelparous species that is vulnerable to endangered in parts of its native range due in part to loss of spawning habitat because of man-made barriers. The ability of lampreys to return to the ocean or estuary and search out alternative spawning river systems would be limited by their osmoregulatory ability in seawater. A reduction in tolerance to salinity has been documented in migrants, although the underlying mechanisms have not been characterized. We examined the capacity for marine osmoregulation in upstream spawning migrants by characterizing the physiological effects of salinity challenge from a molecular perspective. Estuarine-captured migrants held in freshwater (FW) for ∼1 week (short-term acclimation) or 2 months (long-term acclimation) underwent an incremental salinity challenge until loss of equilibrium occurred and upper thresholds of 25 and 17.5, respectively, occurred. Regardless of salinity tolerance, all lamprey downregulated FW ion-uptake mechanisms [gill transcripts of Na(+):Cl(-) cotransporter (NCC/slc12a3) and epithelial Na(+) channel (ENaC/scnn1) and kidney Na(+)/K(+)-ATPase (NKA) protein and activity but not transcript]. At their respective salinity limits, lamprey displayed a clear osmoregulatory failure and were unable to regulate [Na(+)] and [Cl(-)] in plasma and intestinal fluid within physiological limits, becoming osmocompromised. A >90% drop in haematocrit indicated haemolysis, and higher plasma concentrations of the cytosolic enzymes alanine aminotransferase, aspartate aminotransferase and lactate dehydrogenase indicated damage to other tissues, including liver. However, >80% of short-term FW-acclimated fish were able to osmoregulate efficiently, with less haemolysis and tissue damage. This osmoregulatory ability was correlated with significant upregulation of the secretory form of Na(+):K(+):2Cl(-) cotransporter (NKCC1/slc12a2) transcript levels and the re-emergence of seawater-type ionocytes detected through immunohistochemical NKA immunoreactivity in the gill, the central ionoregulatory organ. This work sheds light on the molecular and physiological limits to the potential return to seawater for lampreys searching for alternative FW systems in which to spawn.

Comparative effect of a new mouthrinse containing chlorhexidine, triclosan and zinc on volatile sulphur compounds: a randomized, crossover, double-blind study.

AIM: The aims of this study were to compare the volatile sulphur compounds (VSC)-reducing effect of two commercial mouthrinses using a morning bad breath model and to assess the role of mechanical plaque control (MPC) when performed previously to mouthrinse use. PATIENTS AND METHODS: Eleven volunteers with good oral health were enrolled in a double-blind, randomized, six-step crossover design study with a 7-day washout period. Two commercial mouthrinses were tested using a saline solution (NaCl 0.9%) as a negative control: one mouthrinse contained 0.05% chlorhexidine, 0.05% cetylpyridinium chloride and 0.14% zinc lactate (CHX-CPC-Zn), while the other contained 0.05% chlorhexidine, 0.15% triclosan and 0.18% zinc pidolate (CHX-triclosan-Zn). A portable sulphide monitor (Halimeter(®) ) was used for VSC quantification. Measurements were made at baseline, and 1, 3 and 5 h after rinsing. Significant differences were detected by analysis of variance. RESULTS: No significant differences between groups were detected at baseline. We were unable to demonstrate a significant influence of mechanical plaque control on the reduction of VSC levels when performed before mouthrinse use (P = 0.631). Both mouthrinses effectively lowered VSC levels in all test intervals (P < 0.05). No statistically significant differences were found between mouthrinses in any of the test intervals (P = 0.629, 0.069 and 0.598 at 1, 3 and 5 h). CONCLUSION: This study demonstrated that CHX-CPC-Zn and CHX-triclosan-Zn have significant and similar effects in reducing VSC levels, which persist for at least 5 h. Such effects were independent of previous MPC, which failed to improve on the results of mouthrinse use alone.

Protein-ligand docking in the new millennium--a retrospective of 10 years in the field.

Protein-ligand docking is currently an important tool in drug discovery efforts and an active area of research that has been the subject of important developments over the last decade. These are well portrayed in the rising number of available protein-ligand docking software programs, increasing level of sophistication of its most recent applications, and growing number of users. While starting by summarizing the key concepts in protein-ligand docking, this article presents an analysis of the evolution of this important field of research over the past decade. Particular attention is given to the massive range of alternatives, in terms of protein-ligand docking software programs currently available. The emerging trends in this field are the subject of special attention, while old established docking alternatives are critically revisited. Current challenges in the field of protein-ligand docking such as the treatment of protein flexibility, the presence of structural water molecules and its effect in docking, and the entropy of binding are dissected and discussed, trying to anticipate the next years in the field.

Plasma steroid hormone levels in female flounder Platichthys flesus and the influence of fluctuating hydrostatic pressure.

The reproductive cycle in teleosts is timed to guarantee that eggs hatch in the right place at the right time, with environmental factors playing important roles in entraining and controlling the entire process. The effects of some environmental factors, like temperature and photoperiod, are now well understood. There are only a few studies regarding the effects of hydrostatic pressure (HP) on the reproductive cycle, in spite of its importance as a ubiquitous factor in all biological environments and affecting all living organisms. Hydrostatic pressure is of particular importance in fish because they can also experience rapid and cyclic changes in HP due to vertical movements in the water column. The aim of the present research was to investigate the effects of vertical migrations on the reproductive steroids of maturing female flounder. After a 14 day exposure to cyclic hydrostatic pressure (with a period of 12.4h and with a maximum peak of 800 kPa of absolute hydrostatic pressure), fish showed significantly lower plasmatic concentrations of "5ß,3α" steroids, metabolites of the putative maturation-inducing steroid in flounder (17α,20ß-dihydroxy-4-pregnen-3-one). Results indicate that environmentally realistic cyclic changes of hydrostatic pressure can influence the metabolism of reproductive steroids. This suggests a physiological role of tidally-associated vertical migrations, affecting oocyte maturation and retarding the reproductive cycle in this species until the spawning ground is attained.

The inhibitory effect of environmental ammonia on Danio rerio LPS induced acute phase response.

Ammonia is a toxic by-product of amino acid catabolism and a common environmental pollutant that has been associated with increased disease susceptibility in fish although the mechanism is not well understood. We addressed the hypothesis that elevated environmental ammonia acts by impairing the acute phase response (APR). Specifically, we determined the impact of sub-lethal acute (24 h) and chronic (14 d) ammonia exposure on acute phase protein gene expression in zebrafish (Danio rerio) in response to a challenge with bacterial lipopolysaccharide (LPS: i.p. 10 µg/g after 24h). A panel of LPS-responsive genes (SAA, HAMP, LECT2, Hp and IL1ß) were identified and evaluated by real-time quantitative PCR. Ammonia was found to impair induction of SAA, HAMP and LECT2 by 50-90%. Both short (15 min, 1h and 24h) and long-term (14 days) exposure to high environmental ammonia concentrations significantly elevated whole-body cortisol levels compared with control fish. Our results reveal for the first time that exposure to high environmental levels of ammonia suppresses the innate immune response in fish. We hypothesize that high environmental ammonia-mediated elevation of cortisol levels in zebrafish may be playing a key role in this immunosuppression, while the mechanisms involved remains to be elucidated.

The effect of elevated hydrostatic pressure upon selected biomarkers in juvenile blackspot seabream Pagellus bogaraveo in a 14 day-long experiment.

Hydrostatic pressure elevated to 500 kPa for 14 days was found to affect hepatic 7-ethoxyresorufin-O-deethylase (EROD), oxidized protein (POx), protein yield and branchial Na(+)-K(+)-ATPase. No effect on glutathione-S-transferase (GST), superoxidase dismutase (SOD), catalase (CAT), lipid peroxidation (LP), acetylcholinesterase (AChE), butyrylcholinesterase (BChE), condition factor (K) and hepato-somatic index (I(H)) was encountered.

Water-soluble fraction of mercury, arsenic and other potentially toxic elements in highly contaminated sediments and soils.

The water-soluble contents of mercury, arsenic and other potentially toxic elements in highly contaminated sediment and soil samples from Portugal were determined. Mercury and arsenic concentrations were detectable and reproducible among replicate experiments. Despite the acidic pH, the low organic carbon content and the exceptionally high levels of contamination of certain samples (total mercury contents varied between 0.15 and 3180 mg kg(-1) while total arsenic concentrations ranged from 11 to 6365 mg kg(-1)), the water-soluble percentages of both mercury (<1.2%) and arsenic (<4.6%) were generally low. The variability of the water-soluble fractions of these two elements among these samples and at the occurring pH conditions seems not to be associated with the release of other potentially toxic elements. The highest water-soluble concentrations of the remaining potentially toxic elements were generally observed in the 15-25 cm depth layer of sediments from areas colonised with plants (Halimione portulacoides) and in mining soil samples. Zinc, cobalt, copper and cadmium showed the highest water-soluble percentages of elements in relation to total metal contents. Given the high contamination levels, the availability of potentially toxic elements in these areas as well as possible risks to the environment and humans should be further investigated. The presence of plants (H. portulacoides) appears to cause significant changes in the sediment matrix that increase the mobility of several potentially toxic elements, particularly in the 15-25 cm depth layer. The effects of vegetation on the fractionation of potentially toxic elements on these sediments should be further studied.

Branchial ammonia excretion in the Asian weatherloach Misgurnus anguillicaudatus.

The weatherloach, Misgurnus anguillicaudatus, is a freshwater, facultative air-breathing fish that lives in streams and rice paddy fields, where it may experience drought and/or high environmental ammonia (HEA) conditions. The aim of this study was to determine what roles branchial Na(+)/K(+)-ATPase, H(+)-ATPase, and Rhcg have in ammonia tolerance and how the weatherloach copes with ammonia loading conditions. The loach's high ammonia tolerance was confirmed as was evident from its high 96 h LC(50) value and high tissue tolerance to ammonia. The weatherloach does not appear to make use of Na(+)/NH(4)(+)-ATPase facilitated transport to excrete ammonia when exposed to HEA or to high environmental pH since no changes in activity were observed. Using immunofluorescence microscopy, distinct populations of vacuolar (V)-type H(+)-ATPase and Na(+)/K(+)-ATPase immunoreactive cells were identified in branchial epithelia, with apical and basolateral staining patterns, respectively. Rhesus C glycoprotein (Rhcg1), an ammonia transport protein, immunoreactivity was also found in a similar pattern as H(+)-ATPase. Rhcg1 (Slc42a3) mRNA expression also increased significantly during aerial exposure, although not significantly under ammonia loading conditions. The colocalization of H(+)-ATPase and Rhcg1 to the similar non-Na(+)/K(+)-ATPase immunoreactive cell type would support a role for H(+)-ATPase in ammonia excretion via Rhcg by NH(4)(+) trapping. The importance of gill boundary layer acidification in net ammonia excretion was confirmed in this fish; however, it was not associated with an increase in H(+)-ATPase expression, since tissue activity and protein levels did not increase with high environmental pH and/or HEA. However the V-ATPase inhibitor, bafilomycin, did decrease net ammonia flux whereas other ion transport inhibitors (amiloride, SITS) had no effect. H(+)-ATPase inhibition also resulted in a consequent elevation in plasma ammonia levels and a decrease in the net acid flux. In gill, aerial exposure was also associated with a significant increase in membrane fluidity (or increase in permeability) which would presumably enhance NH(3) permeation through the plasma membrane. Taken together, these results indicate the gill of the weatherloach is responsive to aerial conditions that would aid ammonia excretion.

Effect of cortisol on some osmoregulatory parameters of the teleost, Oreochromis niloticus L., after transference from freshwater to seawater

Este estudo foi realizado com o objectivo de testar os efeitos do cortisol na aclimatação da tilápia Oreochromis niloticus (L.) à água salgada. As tilápias (n=42) foram injectadas intraperitonealmente com cortisol e directamente transferidas de água doce para água salobra (15ë). As alterações da osmolaridade, concentração em cloretos (Cl-), os níveis plasmáticos de cortisol e a actividade branquial da Na+, K+-ATPase foram medidas (6, 12, 24, 48, 72 e 168 horas) após a transferência para água salobra. A osmolaridade e a concentração em Cl- aumentou imediatamente após a transferência até às 12-24h. O grupo injectado com cortisol (F) mostrou níveis plasmáticos de cortisol mais elevados do que o grupo controlo (C) que manteve os níveis iniciais durante a experiência. A actividade branquial da Na+, K+-ATPase dos peixes do grupo C começou às primeiras horas após a transferência e teve um pico às 48h. As diferenças entre a actividade enzimática da Na+, K+-ATPase dos grupos F e C foram significativas (P<0,05) em água doce, o que confirma o efeito exógeno do cortisol.