BAY 41-2272, a soluble guanylate cyclase agonist, activates human mononuclear phagocytes

BACKGROUND AND PURPOSE Phagocyte function is critical for host defense against infections. Defects in phagocytic function lead to several primary immunodeficiencies characterized by early onset of recurrent and severe infections. In this work, we further investigated the effects of BAY 41-2272, a soluble guanylate cyclase (sGC) agonist, on the activation of human peripheral blood monocytes (PBM) and THP-1 cells. EXPERIMENTAL APPROACH THP-1 cells and PBM viability was evaluated by methylthiazoletetrazolium assay; reactive oxygen species production by lucigenin chemiluminescence; gene and protein expression of NAPDH oxidase components by qRT-PCR and Western blot analysis, respectively; phagocytosis and microbicidal activity by co-incubation, respectively, with zymosan and Escherichia coli; and cytokine release by elisa. KEY RESULTS BAY 41-2272, compared with the untreated group, increased spreading of monocytes by at least 35%, superoxide production by at least 50%, and gp91PHOX and p67PHOX gene expression 20 to 40 times, in both PBM and THP-1 cells. BAY 41-2272 also augmented phagocytosis of zymosan particles threefold compared with control, doubled microbicidal activity against E. coli and enhanced the release of TNF-a and IL-12p70 by both PBM and THP-1 cells. Finally, by inhibiting sGC with ODQ, we showed that BAY 41-2272-induced superoxide production and phagocytosis is not dependent exclusively on sGC activation. CONCLUSIONS AND IMPLICATIONS In addition to its ability to induce vasorelaxation and its potential application for therapy of vascular diseases, BAY 41-2272 was shown to activate human mononuclear phagocytes. Hence, it is a novel pro-inflammatory drug that may be useful for controlling infections in the immunocompromised host.

Population biology of Stellifer rastrifer, S. brasiliensis and S. stellifer in Caraguatatuba Bay, northern coast of São Paulo, Brazil

Ecosystems may reflect environmental changes in many respects, from the debilitation of individuals to alterations in community composition. Equally, population parameters may provide reliable indications of environmental changes. Members of the sciaenid fish genus Stellifer are usually very abundant where they occur, often with two or more species living in sympatry. Here, the population dynamics of three Stellifer species from southeastern Brazil were assessed. Sampling was carried out in shallow marine areas of Caraguatatuba Bay, from August 2003 to October 2004. The species evaluated were Stellifer rastrifer (n=3183), S. brasiliensis (n=357) and S. stellifer (n=116). The area under greater continental influence tended to support more, but smaller individuals. Size variations over time were similar among species and negatively correlated with Krel, which showed smooth fluctuations. The general length-frequency distribution was concentrated between 6.0 and 9.0 cm, and the great majority of females did not present mature gonads during the sampling period. The findings support the existence of a stratification by size for these species, indicating that the area is essential for the development of younger fish. Failure to consider these characteristics for the management of similar areas may have serious implications for these environments.

Population biology and diet of the southern kingcroaker Menticirrhus americanus (Linnaeus, 1758) (Perciformes: Sciaenidae) in Caraguatatuba Bay, southeastern Brazil

This study evaluated the spatio-temporal distribution, population biology and diet of Menticirrhus americanus in Caraguatatuba Bay. Samples were taken monthly between August 2003 and October 2004, by trawling in two previously selected areas. The northern area is more exposed to wave activity and is influenced by a river, functioning as a small estuary. In contrast, the southern area is relatively sheltered from wave energy and influenced to a lesser degree by smaller rivers. The fishes' length was measured, and the sex and gonadal stage macroscopically identified. The abundance of this species was compared between areas and among months. The diet was identified and quantified. M. americanus occurred in equal proportions in the two study areas, being most abundant in April 2004, followed by December 2003 and January 2004. The population was dominated by small immature individuals. The few individuals in maturation or mature that were captured showed no seasonal pattern of distribution. This species had a varied diet, feeding on worms (nemerteans, sipunculans and echiurans), mollusks (bivalves and cephalopods), polychaetes, crustaceans and fish. The presence of intact nematodes in the intestine suggests that these are parasites. The results demonstrated that M. americanus has a homogeneous spatial and temporal distribution in Caraguatatuba Bay, being uniformly distributed between the south and north areas as well as across the months. This species can be considered a carnivorous predator, showing a preference for consuming benthic sandy-beach species such as glycerids and other polychaetes, crustaceans, and bivalve siphons.

Population biology of the barbel drum Ctenosciaena gracilicirrhus (Metzelaar, 1919) (Perciformes: Sciaenidae) in Caraguatatuba Bay, Southeastern Brazil

This study evaluated the population biology of Ctenosciaena gracilicirrhus (Perciformes: Sciaenidae) in a shallow non-estuarine coastal area of southeastern Brazil. Monthly samples were taken from October 2003 through October 2004, in two distinct areas at depths from 1 to 4 m. C. gracilicirrhus was generally among the most abundant fish species during the period. Its density was significantly higher in a single sampling month, May 2004, in the South area, which may be explained by its migratory behavior together with its preference for deeper areas. Such behavior may lead to bias in community estimates that use few or only seasonal samples. C. gracilicirrhus individuals ranged from 4.0 to 10.2 cm long, with a main mode from 7.5 to 9.0 cm and a significant decrease in mean size from June onward. The estimation of body growth parameters was compromised by this population feature. Similarly, the prey spectrum was difficult to determine because of the deteriorated condition of the stomach contents, although crustaceans were clearly the most important items ingested. Amphipoda was the only subgroup that could be identified more precisely, mainly by the construction of their tubes.

Catch rates of artisanal fisheries targeting Dasyatis guttata (Bloch & Schneider, 1801) in Todos os Santos Bay, northeastern Brazil

Dasyatis guttata has been target of artisanal fisheries in the coast of Bahia (Northeast Brazil) mainly by “arraieira” (gillnet) and “grozeira” (bottom long-line), but until now there is no stock assessment study. One of the important data for this knowledge is reliable indices of abundance. The aims of the present work are to: (1) estimate the best predictor for relative abundance (catch-per-unit-of-effort, CPUE), examining whether catch (production – kg) was related to: soak time of the gear, size of the gillnet or number of hooks, applying generalized linear model (GLM); (2) estimate the annual CPUE (kg/hooks and kg/m) averaged by gear; and (3) assess the temporal CPUE variance. Based on monthly sampling between January 2012 and January 2013, 222 landings by grozeira and 76 by arraiaiera were recorded in the two landing sites in Todos os Santos Bay, Bahia. A total of 14,550 kg (average = 44 kg/month) of D. guttata was captured. Models for both gears were highly significant (P < 0.0001). The analysis indicated that the most appropriate variable for CPUE analysis was the size of the gillnet (P < 0.001) and the number of hooks (P < 0.0001). Soak time of the gear was not significant for both gears (P = 0.4). High residual deviance expresses the complexity of the relations between ecosystem factors and other fisheries factors affecting relative abundance, which were not considered in this study. The average CPUE by grozeira was 6.39 kg/100 hooks ± 8.89 and by arraieira, 1.47 kg/100 m ± 1.66 over the year. Kruskal-Wallis test showed effect of the month on the mean grozeira CPUE (P = <0.001), but no effect (P = 0.096) on the mean arraieira CPUE. Grozeira CPUE values were highest in December and March, and lowest between May to August

Application of 137Cs and 210Pb radionuclides to determine sedimentation rates of recent sediments from admiraty Bay, Antarctica Peninsula

Studies about natural and artificial radionuclides in areas such as the Antarctic are key to understand natural and dynamic processes in marine environments. These studies are important to determine levels of radioactive elements and local sedimentation rates. Five marine sediment cores were collected in different points of Admiralty Bay, in the Antarctic Peninsula. The purpose of this study was to determine 137Cs, 226Ra and 210Pb and sedimentation rates at each site. 137Cs, 210Pb and 226Ra were assayed by gamma-counting through direct measurement of the peak at 661 keV, 47 keV and 609 keV, respectively. Sedimentation rates were obtained by 137Cs and 210Pb (CIC and CRS). The activities for 137Cs ranged from 0.84 to 7.09 Bq kg-1; to 226Ra from 6.77 to 31.07 Bq kg-1 and for 210Pb ranged from 1.10 to 36.90 Bq kg-1. The sedimentation rates obtained by the three models ranged from 0.11±0.01 cm y-1 to 0.46±0.05 cm y-1. The levels of 137Cs registered in this study, as well as in other studies in the Antarctic region indicate that global fallout is the main cause of artificial radionuclides present in this environment, since the Antarctic has not suffered a direct action of human activities that released radioactive elements. The possible grain size variations that occur in the studied points of Admiralty Bay may explain the differences found in the vertical distribution of radionuclides, because of the different values of sedimentation rates and respective dating determined in their profiles

Mesozooplankton community structure during summer months in the Bay of Cádiz

[EN] Mesozooplankton organisms (>250 μm) were sampled at two stations (inner and outer Bay) in the Bay of Cádiz between May and July 2008. Samples were analysed by means of a semi-automated technique in order to give a preliminary view of the mesoozooplankton community structure in the Bay, based on taxonomic diversity and biomass distribution among size classes. The abundance of organisms increased from May to July in accordance with the increase in temperature and Chlorophyll a (Chla) concentrations. Abundances were higher in the outer Bay station, where Chla concentrations are greater and the water column is more stable. The community changed from being meroplankton- to holoplankton-based due to an increase of Calanoida and especially Cladocera individuals (mainly Peniliaavirostris), which are known to peak acutely in the summer. The analysis of Normalised Biomass-Size spectra revealed fairly steep slopes (average -1.3) and relatively high departures from steady state (r2 = 0.8 – 0.94), expectable in a coastal system such as the Bay of Cádiz were disturbance factors are introduced from benthic and tidal processes, together with anthropogenic pressure.

Mikrosomale Biotransformation von Benzo[ghi]perylen, einem mutagenen polyaromatischen Kohlenwasserstoff ohne das Strukturelement der Bay-Region des kanzerogenen Benzo[a]pyrens

Kanzerogene polyaromatische Kohlenwasserstoffe (PAKs), wie Benzo[a]pyren, besitzen eine Bay-Region mit ortho-kondensiertem Benzoring. Dadurch ist die enzymatische Bildung von Bay-Region-Dihydrodiolepoxiden (Oxiranylring in der sterisch abgeschirmten Molekülbucht) möglich, die als ultimal kanzerogene Metaboliten der PAKs gelten. Diese lösen durch DNA-Modifikation Primärläsionen aus, die, sofern sie nicht enzymatisch repariert werden, bei der DNA-Replikation Fehler verursachen (Mu-tationen). Der Mehrstufenprozeß der Kanzerogenese (Promotion und Progression) führt schließlich zur neoplastischen Entartung der Zelle. Benzo[ghi]perylen (BghiP) repräsentiert eine Gruppe von PAKs, die keine „klassische“ Bay-Region besitzen und daher keine vicinalen Dihydrodiolepoxiden bilden können. Trotzdem ist BghiP mutagen, z. B. in den Stämmen TA98 und TA100 von Salmonella typhimurium (1,3- bzw. 4,3 his+-Revertanten/nmol) nach metabolischer Aktivierung mit der postmitochondrialen Fraktion von Ratten nach Behandlung mit 3-Methylcholanthren. Hemmung der mikrosomalen Epoxidhydrolase (mEH) mit 1,1,1-Trichlor-2-propenoxid (TCPO) steigert die bakterielle Mutagenität von BghiP im Stamm TA98 um das 4-fache, was Arenoxide als ultimale Mutagene wahrscheinlich macht. Dieses Ergebnis wird au-ßerdem durch Untersuchung der DNA-Bindung mit dem Verfahren des 32P-Postlabelings bestätigt (Dr. Fickler, Institut für Toxikologie, Universität Mainz). Danach bildete mikrosomal aktiviertes BghiP drei Addukte (ein Hauptaddukt, zwei Nebenaddukte), die durch Hemmung der mEH mit TCPO verstärkt wurden (das Hauptaddukt um 29%). Um den für die bakterielle Mutagenität von BghiP verantwortlichen Metaboliten zu identifizieren, wurde die mikrosomale Biotransformaton von BghiP aufgeklärt. Umsetzung von BghiP mit Lebermikrosomen von Ratten nach Behandlung mit Aroclor 1254 lieferte 17 mit Ethylacetat extrahierbare Metaboliten. Zwölf dieser Metaboliten konnten durch eine Kombination von chromatographischen, spektroskopi-schen und biochemischen Methoden identifiziert werden. Daraus ergeben sich zwei Biotransformati-onswege: Weg I beginnt mit einem Angriff von Cytochrom P450-abhängigen Monooxygenasen an Position 7 und der Bildung des 7-Phenols. Dieses wird dann in das 7,8- bzw. 7,10-Diphenol überführt, die schließlich zu den mehrkernigen Chinonen an der 7,8- bzw. 7,10-Position oxidiert werden. Im Bio-transformationsweg II werden die K-Regionen von BghiP durch Cytochrom P450 funktionalisiert. Zu-nächst entstehen das auf indirektem Weg identifizierte 3,4-Oxid und das 3,4,11,12-Bisoxid, die in mikrosomalen Umsetzungen von BghiP nur nach Hemmung der mEH gebildet werden. Enzymatische Hydrolyse des 3,4-Oxides ergibt das trans-3,4-Dihydrodiol, das zum 3,4-Chinon oxidiert wird. Ebenso entsteht aus dem 3,4,11,12-Bisoxid das trans-3,4-trans-11,12-Bisdihydrodiol, aus dem durch Oxidati-on das trans-3,4-Dihydrodiol-11,12-Chinon hervorgeht. Untersuchung der stereoselektiven enzymati-schen Bildung der K-Region-trans-Di¬hydrodiole ergaben eine präferentielle Entstehung der 3R,4R- bzw. 3R,4R,11R,12R-Enantiomere. Untersuchungen der bakteriellen Mutagenität der Hauptmetaboliten 3,4-Dihydrodiol und dem 7-Phenol machte deutlich, dass beide Biotransformationswege I und II von BghiP zur bakteriellen Mutagenität beitragen. Das 7-Phenol aus Weg I ist ein proximales Mutagen, was auch von Phenolen anderer PAKs bekannt ist. Das 3,4-Dihydrodiol aus Weg II wird so schwach zu Mutagenen aktiviert, dass dem vermutlich gebildete 3,4-Dihydrodiol-11,12-oxid keine große Bedeutung als ultimales Mutagen von BghiP zukommt. Die Bestimmung der direkten mutagenen Aktivität (ohne metabolische Aktivierung) der mutmaßlich ultimal mutagenen Arenoxide von BghiP ergab, dass die des 3,4,11,12-Bisarenoxides sehr gering war (1,3 his+-Revertanten/nmol im Stamm TA98). Das 3,4-Oxid hingegen bewirkte einen deutlichen gentoxischen Effekt in den Stämmen TA98 und TA100 (5,5 bzw. 10 his+-Revertanten/nmol). Dies wurde durch die Bestimmung der DNA-Bindung mit dem 32P-Postlabeling, in dem das 3,4-Oxid für das Hauptaddukt von BghiP verantwortlich gemacht werden konnte, bestätigt. Daher kommt dem 3,4-Oxid als ultimales Mutagen die größte Bedeutung für die Gentoxizität von BghiP zu. Die Ergebnisse dieser Arbeit lassen bei PAKs ohne Bay-Region auf Arenoxide schließen, die eine notwendige Voraussetzung für DNA-Bindung und Mutagenität sind.

Potential effect of marine traffic on white-beaked dolphin (Lagenorhynchus albirostris) distribution and behaviour in Faxaflói Bay, southwest Iceland

Faxaflói bay is a short, wide and shallow bay situated in the southwest of Iceland. Although hosting a rather high level of marine traffic, this area is inhabited by many different species of cetaceans, among which the white-beaked dolphin (Lagenorhynchus albirostris), found here all year-round. This study aimed to evaluate the potential effect of increasing marine traffic on white-beaked dolphins distribution and behaviour, and to determine whether or not a variation in sighting frequencies have occurred throughout years (2008 – 2014). Data on sightings and on behaviour, as well as photographic one, has been collected daily taking advantage of the whale-watching company “Elding” operating in the bay. Results have confirmed the importance of this area for white-beaked dolphins, which have shown a certain level of site fidelity. Despite the high level of marine traffic, this dolphin appears to tolerate the presence of boats: no differences in encounter durations and locations over the study years have occurred, even though with increasing number of vessels, an increase in avoidance strategies has been displayed. Furthermore, seasonal differences in probabilities of sightings, with respect to the time of the day, have been found, leading to suggest the existence of a daily cycle of their movements and activities within the bay. This study has also described a major decline in sighting rates throughout years raising concern about white-beaked dolphin conservation status in Icelandic waters. It is therefore highly recommended a new dedicated survey to be conducted in order to document the current population estimate, to better investigate on the energetic costs that chronic exposure to disturbances may cause, and to plan a more suitable conservation strategy for white-beaked dolphin around Iceland.