Inhibition of Na+,K+-ATPase by Ouabain Opens Calcium Channels Coupled to Acetylcholine Release in Guinea Pig Myenteric Plexus

Abstract: Ouabain, an Na⁺,K⁺-ATPase inhibitor, increases the release of acetylcholine (ACh) from various preparations in a Ca²⁺-independent way. However, in other preparations the release of ACh evoked by ouabain is dependent on the presence of extracellular calcium. In the present study, we have labeled the ACh of myenteric plexus longitudinal muscles of guinea pig ileum and compared the effect of calcium channel blockers on ouabain-evoked release of [³H]ACh. Release of [³H]ACh evoked by ouabain is dose dependent and decreased markedly in the absence of calcium or in the presence of cadmium, a nonspecific calcium channel blocker. N-type calcium channel blockage by the ω-conotoxins GVIA (selective N-type calcium channel blocker) and MVIIC (a nonselective calcium channel blocker) inhibited by 45 and 55%, respectively, the release of [³H]ACh. L-type calcium channel suppression by low concentrations of verapamil, nifedipine, and diltiazem had no effect on the release of [³H]ACh. The release of transmitter was also not affected significantly by nickel, a T-type calcium channel blocker. In addition, ω-agatoxin-IVA, at concentrations that block P- and Q-type calcium channels, did not affect significantly the release of [³H]ACh. Thus, extracellular Ca²⁺ is essential for the release of ACh induced by ouabain from guinea pig ileum myenteric plexus. In this preparation, the N-type calcium channel plays a dominant role in transmitter release evoked by inhibition of Na⁺,K⁺-ATPase, but other routes of calcium entry in addition to these channels can also support the release of neurotransmitter induced by ouabain.     

Follower fish of the goldspotted eel <Emphasis Type="Italic">Myrichthys ocellatus</Emphasis> with a review on anguilliform fish as nuclear species

In a nuclear-follower fish foraging association, the follower benefits from food uncovered or flushed out when the nuclear fish disturbs the bottom, while nuclear species generally do not seem to be benefiting. Among nuclear species, eels (anguilliform fish) are known to be one of the most represented groups. Here we investigated the frequency and time duration of foraging associations among the goldspotted eel Myrichthys ocellatus and reef fish in a subtropical marginal reef. In addition, we reviewed nuclear eel species and their followers described in the literature. From a total of 211 goldspotted eels observed, seven follower species were recorded in 19% of the samples. The average time of the following associations per species ranged from 40 to 190 s. Four species were reported for the first time as M. ocellatus followers (Bodianus rufus, B. pulchellus, Stephanolepis hispidus, and Serranus baldwini) and three of them have never been reported in the literature as eel followers (B. pulchellus, S. hispidus, and S. baldwini). The literature describes 13 eel species acting as nuclear for 66 fish species, represented mainly by groupers and sea basses. The size of the eel was not correlated with the size of its follower and neither with the number of described follower species. The nuclear role of eels is likely to be an important component of the trophic ecology of small and medium-sized macrocarnivore fish.     

Prostate cancer proteomics: clinically useful protein biomarkers and future perspectives

Introduction: Although prostate cancer constitutes one of the most important, death-related diseases in the male population, there is still a need for identification of sensitive biomarkers that could precociously detect the disease and differentiate aggressive from indolent cancers, in order to decrease overtreatment. Proteomics research has improved understanding on mechanisms underlying tumorigenesis, cancer cells migration and invasion potential, and castration resistance. This review has focused on proteomic studies of prostate cancer published in the recent years, with a special emphasis on determination of biomarkers for cancer progression and diagnosis.

Areas covered: Shotgun and targeted-proteomic studies of prostate cancer in different matrices are reviewed, i.e., prostate tissue, prostate cell lines, blood (serum and plasma), urine, seminal plasma, and exosomes. The most important biomarkers for cancer diagnosis and aggressiveness characterization are highlighted.

Expert commentary: In general, results demonstrate alteration in cell cycle control, DNA repair, proteasomal degradation, and metabolic activity. However, these studies suffer from low reproducibility due to heterogeneity of the cancer itself, as well as to techniques utilized for protein identification/quantification. Downstream confirmatory studies in separate cohorts are warranted in order to demonstrate accuracy of these results.     

A review of the biology,ecology, behavior and conservation status of the dusky grouper, <Emphasis Type="Italic">Epinephelus marginatus</Emphasis> (Lowe 1834)

The dusky grouper is a large-bodied marine species usually associated with rocky substrates and reefs currently experiencing increasing anthropogenic pressures. Here, we critically evaluate studies conducted during the last few decades on this endangered species, focusing on its biology, ecology, behavior, stock structure and fisheries, and identify future research directions to fill current knowledge gaps. This species plays an important functional role in the rocky bottom habitats in which it lives, usually occupying the highest trophic levels and with the potential role as a keystone species in some ecosystems. It has a complex life cycle characterized by a high longevity, a slow growth rate and a monandric protogynous hermaphroditic mode of reproduction. The species also exhibits spawning aggregation behavior, which makes it more vulnerable to fishing activity. Aside from its ecological importance, this species has high commercial value for professional fishers and is also highly prized by recreational spearfishers. These biological features, associated with increasing fishing pressures throughout most of its geographical distribution, have led to Epinephelus marginatus being included since 2004 in the IUCN Red List of Threatened Species as Endangered. Despite a large amount of research being conducted on this species over the last five decades or so, using a number of different tools and techniques, key questions regarding its biology still remain unanswered. In terms of conservation and management plans designed to safeguard this endangered species, we identified the crucial need to improve fisheries landing statistics for the species and to increase the number of marine protected areas within its distribution beyond the Mediterranean Sea.     

Overdispersed fungus germination data: statistical analysis using R

ABSTRACT

Proportion data from dose-response experiments are often overdispersed, characterised by a larger variance than assumed by the standard binomial model. Here, we present different models proposed in the literature that incorporate overdispersion. We also discuss how to select the best model to describe the data and present, using R software, specific code used to fit and interpret binomial, quasi-binomial, beta-binomial, and binomial-normal models, as well as to assess goodness-of-fit. We illustrate applications of these generalized linear models and generalized linear mixed models with a case study from a biological control experiment, where different isolates of Isaria fumosorosea (Hypocreales: Cordycipitaceae) were used to assess which ones presented higher resistance to UV-B radiation. We show how to test for differences between isolates and also how to statistically group isolates presenting a similar behaviour.     

Voltages up to 600V did not affect cryopreserved bovine spermatozoa on capillary-type electroporation

Physical methods such as electroporation have been used to improve the DNA uptake efficiency of sperm cells. This study aims to develop an efficient capillary-type electroporation method for incorporation of exogenous DNA into bovine cryopreserved sperm cells with minimal detrimental effects for later use in SMGT. Electroporation of the samples was performed in 2 different groups (with 1?μg of DNA and without DNA transfection) and under five different voltages: 500?V, 600?V, 700?V, 800?V and 900?V. Non-electroporated sperm cells (with and without DNA) were used as control. Kinetics parameters were determined using computer assisted semen analyses, whereas membrane integrity, fluidity, mitochondrial function and DNA uptake were evaluated by flow cytometry. Results revealed that all tested voltages reduced electroporated sperm motility (P?<?0.05) when compared to the control (non-electroporated cells). Mitochondrial function results showed no statistical difference among groups. Similarly, groups electroporated with lower (500?V, 600?V and 700?V) voltages showed no difference in cell membrane integrity and fluidity. Groups electroporated at higher voltages (800?V and 900?V) demonstrated negative effects in cells membrane integrity when compared to other groups and control. Also, all electroporated groups demonstrated significant higher percentages of transfected sperm cells when compared to the control group (P?<?0.05). Under the recommendation of using voltages up to 600?V, this method represents a safe and efficient alternative for electroporation of bovine spermatozoa.     

Bacteria associated with Amblyomma cajennense tick eggs

Ticks represent a large group of pathogen vectors that blood feed on a diversity of hosts. In the Americas, the Ixodidae ticks Amblyomma cajennense are responsible for severe impact on livestock and public health. In the present work, we present the isolation and molecular identification of a group of culturable bacteria associated with A. cajennense eggs from females sampled in distinct geographical sites in southeastern Brazil. Additional comparative analysis of the culturable bacteria from Anocentor nitens, Rhipicephalus sanguineus and Ixodes scapularis tick eggs were also performed. 16S rRNA gene sequence analyses identified 17 different bacterial types identified as Serratia marcescens, Stenotrophomonas maltophilia, Pseudomonas fluorescens, Enterobacter spp., Micrococcus luteus, Ochrobactrum anthropi, Bacillus cereus and Staphylococcus spp., distributed in 12 phylogroups. Staphylococcus spp., especially S. sciuri, was the most prevalent bacteria associated with A. cajennense eggs, occurring in 65% of the samples and also frequently observed infecting A. nitens eggs. S. maltophilia, S. marcescens and B. cereus occurred infecting eggs derived from specific sampling sites, but in all cases rising almost as pure cultures from infected A. cajennense eggs. The potential role of these bacterial associations is discussed and they possibly represent new targets for biological control strategies of ticks and tick borne diseases.     

Nurse abundance determines plant facilitation networks of subtropical forest–grassland ecotone

The theory of complex networks has been recently used to explain ecological associations between nurses and seedlings in plant facilitation systems. The structure of these networks is potentially affected by morphological, ecological and evolutionary factors that can determine the facilitative interactions. In the present study, we evaluate the role of the projected crown area of the nurse plant, the dispersal syndrome, the abundance and the phylogeny relationships with seedlings as drivers of network structure in facilitation networks. To test these parameters, we used an ecotonal forest–grassland system in southern Brazil that experienced historical forest expansion over the grassland in the last few centuries. In two State Parks, Guartelá (GUA) and Vila Velha (VVL), we sampled tree species (nurses and seedlings) along four transects parallel to the forest edge, with five sampling stations along each transect. Tree seedlings (height >0.3 m) were sampled below the nurse's crown (i.e. isolated woody plants, height >1.5 m) and over the grassland area. A total of 160 nurses and 358 seedlings were sampled. Seedling abundance and richness were greater below a nurse's crown than on open grassland. Nurse abundance was the best predictor of the observed interaction matrix. Probabilistic matrices based on abundance, abundance and phylogeny, and dominance were able to predict the observed nestedness values, and they were also closest to the observed connectance, although all of the probabilistic matrices have underestimated this property. Specialization was predicted by abundance and phylogeny, abundance and projected crown area drivers for VVL, and the abundance based models were closest to the predicted specialization for GUA. The result of a similar pattern in predictability between sites indicates that the influences of morphology and evolutionary and ecological processes over the facilitation interactions are equivalent on a regional scale. Woody plant abundance was a key factor for the facilitation networks, driving forest expansion along the subtropical forest–grassland ecotone.