Characterization and modulation of microglial phenotypes in an animal model of severe sepsis

We aim to characterize the kinetics of early and late microglial phenotypes after systemic inflammation in an animal model of severe sepsis and the effects of minocycline on these phenotypes. Rats were subjected to CLP, and some animals were treated with minocycline (10 ug/kg) by i.c.v. administration. Animals were killed 24 hours, 5, 10 and 30 days after sepsis induction, and serum and hippocampus were collected for subsequent analyses. Real‐time PCR was performed for M1 and M2 markers. TNF‐α, IL‐1β, IL‐6, IL‐10, CCL‐22 and nitrite/nitrate levels were measured. Immunofluorescence for IBA‐1, CD11b and arginase was also performed. We demonstrated that early after sepsis, there was a preponderant up‐regulation of M1 markers, and this was not switched to M2 phenotype markers later on. We found that up‐regulation of both M1 and M2 markers co‐existed up to 30 days after sepsis induction. In addition, minocycline induced a down‐regulation, predominantly, of M1 markers. Our results suggest early activation of M1 microglia that is followed by an overlap of both M1 and M2 phenotypes and that the beneficial effects of minocycline on sepsis‐associated brain dysfunction may be related to its effects predominantly on the M1 phenotype.     

Guanosine,a guanine-based nucleoside relaxed isolated corpus cavernosum from mice through cGMP accumulation

In corpus cavernosum (CC), guanosine triphosphate (GTP) is converted into cyclic guanosine monophosphate (cGMP) to induce erection. The action of cGMP is terminated by phosphodiesterases and efflux transporters, which pump cGMP out of the cell. The nucleotides, GTP, and cGMP were detected in the extracellular space, and their hydrolysis lead to the formation of intermediate products, among them guanosine. Therefore, our study aims to pharmacologically characterize the effect of guanosine in isolated CC from mice. The penis was isolated and functional and biochemical analyses were carried out. The guanine-based nucleotides GTP, guanosine diphosphate, guanosine monophosphate, and cGMP relaxed mice corpus cavernosum, but the relaxation (90.7?±?12.5%) induced by guanosine (0.000001–1 mM) was greater than that of the nucleotides (~?45%, P?<?0.05). Guanosine-induced relaxation was not altered in the presence of adenosine type 2A and 2B receptor antagonists. No augment was observed in the intracellular levels of cyclic adenosine monophosphate in tissues stimulated with guanosine. Inhibitors of nitric oxide synthase (L-NAME, 100 μM) and soluble guanylate cyclase (ODQ, 10 μM) produced a significant reduction in guanosine-induced relaxation in all concentrations studied, while in the presence of tadalafil (300 nM), a significant increase was observed. Pre-incubation of guanosine (100 μM) produced a 6.6-leftward shift in tadalafil-induced relaxation. The intracellular levels of cGMP were greater when CC was stimulated with guanosine. Inhibitors of ecto-nucleotidases and xanthine oxidase did not interfere in the response induced by guanosine. In conclusion, our study shows that guanosine relaxes mice CC and opens the possibility to test its role in models of erectile dysfunction.

     

The renin–angiotensin system plays a major role in voiding dysfunction of ovariectomized rats

Aims

The renin–angiotensin system (RAS) plays a major role in cardiovascular diseases in postmenopausal women, but little is known about its importance to lower urinary tract symptoms. In this study we have used the model of ovariectomized (OVX) estrogen-deficient rats to investigate the role of RAS in functional and molecular alterations in the urethra and bladder.

Main methods

Responses to contractile and relaxant agents in isolated urethra and bladder, as well as cystometry were evaluated in 4-month OVX Sprague–Dawley rats. Angiotensin-converting enzyme activity and Western blotting for AT1/AT2 receptors were examined.

Key findings

Cystometric evaluations in OVX rats showed increases in basal pressure, capacity and micturition frequency, as well as decreased voiding pressure. Angiotensin II and phenylephrine produced greater urethral contractions in OVX compared with Sham group. Carbachol-induced bladder contractions were significantly reduced in OVX group. Relaxations of urethra and bladder to sodium nitroprusside and BAY 41-2272 were unaffected by OVX. Angiotensin-converting enzyme activity was 2.6-fold greater (p < 0.05) in urethral tissue of OVX group, whereas enzyme activity in plasma and bladder remained unchanged. Expressions of AT1 and AT2 receptors in the urethra were markedly higher in OVX group. In bladder, AT1 receptors were not detected, whereas AT2 receptor expression was unchanged between groups. 17β-Estradiol replacement (0.1 mg/kg, weekly) or losartan (30 mg/kg/day) largely attenuated most of the alterations seen in OVX group.

Significance

Prolonged estrogen deprivation leads to voiding dysfunction and urethral hypercontractility that are associated with increased ACE activity and up-regulation of angiotensin AT1/AT2 receptor in the urethral tissue.     

Relative efficiency of the genotypic value and combining ability effects on reciprocal recurrent selection

Reciprocal recurrent selection (RRS) has been successfully applied to maize breeding for more than 60 years. Our objective was to assess the relative efficiency of the genotypic value and the effects of general and specific combining abilities (GCA and SCA) on selection. The GCA effect reflects the number of favorable genes in the parent. The SCA effect primarily reflects the differences in the gene frequencies between the parents. We simulated three traits, three classes of populations, and 10 cycles of half- and full-sib RRS. The RRS is a highly efficient process for intra- and interpopulation breeding, regardless of the trait or the level of divergence among the populations. The RRS increases the heterosis of the interpopulation cross when there is dominance, and it decreases the inbreeding depression in the populations and the genetic variability in the populations and in the hybrid. When there is not dominance and the populations are not divergent, the RRS also determines population differentiation. The half-sib RRS, which is equivalent to selection based on the GCA effect, is more efficient than the full-sib RRS based on the genotypic value, regardless of the trait or the level of improvement of the populations. The full-sib RRS based on the SCA effect is not efficient for intra- and interpopulation breeding.     

Vatairea macrocarpa Lectin (VML) Induces Depressive-like Behavior and Expression of Neuroinflammatory Markers in Mice

Lectins are proteins capable of reversible binding to the carbohydrates in glycoconjugates that can regulate many physiological and pathological events. Galectin-1, a β-galactoside-binding lectin, is expressed in the central nervous system (CNS) and exhibits neuroprotective functions. Additionally, lectins isolated from plants have demonstrated beneficial action in the CNS. One example is a lectin with mannose-glucose affinity purified from Canavalia brasiliensis seeds, ConBr, which displays neuroprotective and antidepressant activity. On the other hand, the effects of the galactose-binding lectin isolated from Vatairea macrocarpa seeds (VML) on the CNS are largely unknown. The aim of this study was to verify if VML is able to alter neural function by evaluating signaling enzymes, glial and inflammatory proteins in adult mice hippocampus, as well as behavioral parameters. VML administered by intracerebroventricular (i.c.v) route increased the immobility time in the forced swimming test (FST) 60 min after its injection through a carbohydrate recognition domain-dependent mechanism. Furthermore, under the same conditions, VML caused an enhancement of COX-2, GFAP and S100B levels in mouse hippocampus. However, phosphorylation of Akt, GSK-3β and mitogen-activated protein kinases named ERK1/2, JNK1/2/3 and p38^MAPK, was not changed by VML. The results reported here suggest that VML may trigger neuroinflammatory response in mouse hippocampus and exhibit a depressive-like activity. Taken together, our findings indicate a dual role for galactose binding lectins in the modulation of CNS function.     

Shifts in the dominance between diatoms and cryptophytes during three late summers in the Bransfield Strait (Antarctic Peninsula)

Recent global warming reduces surface water salinity around the Antarctic Peninsula as a result of the glacial meltwater runoff, which increases the occurrence and abundance of certain phytoplankton groups, such as cryptophytes. The dominance of this particular group over diatoms affects grazers, such as Antarctic krill, which preferentially feed on diatoms. Using three late summer data sets from the Bransfield Strait (2008–2010), we observed variations in the dominant phytoplankton groups determined by HPLC/CHEMTAX pigment analysis and confirmed by microscopy. Results indicate that the dominance of diatoms, particularly in 2008 and 2009, was associated with a deeper upper mixed layer (UML), higher salinity and warmer sea surface temperature. In contrast, cryptophytes, which were dominant in 2010, were associated with a shallower UML, lower salinity and colder sea surface temperatures. The low diatom biomass observed in the summer of 2010 was associated with high nutrient concentration, particularly silicate, and low chlorophyll a (summer monthly average calculated from satellite images). The interannual variability here observed suggests a delayed seasonal succession cycle of phytoplankton in the summer of 2010 associated with a cold summer and a late ice retreat process in the region. This successional delay resulted in a notable decrease of primary producers’ biomass, which is likely to have impacted regional food web interactions. This study demonstrates the susceptibility of the Antarctic phytoplankton community structure to air temperature, which directly influences the timing of ice melting and consequently the magnitude of primary production and succession pattern of phytoplankton groups.     

Evidence that human and equine erythrocytes could have significant roles in the transport and delivery of amino acids to organs and tissues

Amino Acids - Erythrocytes have a well-defined role in the gaseous exchange of oxygen and carbon dioxide in the mammalian body. The erythrocytes can contain more than half of the free amino acids...     

Phenology,pollination, and breeding system of the threatened tree Caesalpinia echinata Lam. (Fabaceae), and a review of studies on the reproductive biology in the genus

Caesalpinia echinata (brazilwood) is a threatened tree endemic to the Brazilian Atlantic forest with a global importance as it is worldwide used to manufacture the most desirable bows for violins, violas, and cellos. Although there is an international initiative for its conservation, there is a lack of studies on the reproductive biology of the species, as for the genus, a fundamental understanding for the conservation of any species. In this paper, we investigated the phenology, pollination, and breeding system of C. echinata, presenting some conservation guidelines for the species, together with a review of studies on the reproductive biology in Caesalpinia s.l. The genus has a complex taxonomic history with recent attempts to reassign its limits. Pending a final resolution of the placing of some species (including C. echinata) our revision covers the traditional Caesalpinia s.l. and includes annotations indicating to which segregate genus each species is now considered to belong, as well as the appropriate combination where this has already been published. Field work was undertaken from October/2004 to December/2006 at the Tapacurá Ecological Station, northeastern Brazil. Flowering occurred mainly in the dry season and the peak of seed dispersal was at the beginning of the wet season. Anthesis is diurnal, lasting 1 day. The flowers are zygomorphic, yellow, sweet-scented, and the effective pollinators were mainly medium-sized to large bees of the genera Centris and Xylocopa, together with the introduced Apis mellifera. Nectar volume and sugar concentration averaged 2.9±1.0 μL and 29.5±9.4%, respectively. The ovary has 1–4 ovules (2.35±0.58) and the pollen/ovule ratio was 5631.2; pollen viability was high (95.9±4.8%). Natural fruit set was low (9.2%) with mature fruits averaging 1.7±0.9 seeds. Results of the controlled pollinations and analysis of pollen tube growth revealed C. echinata presents late-acting self-incompatibility. The pollination biology and breeding system of C. echinata are discussed, together with available data on its genetics and physiology, in terms of best conservation practice for this endangered species. Data on the reproductive biology of the genus are scarce, revealing the predominance of bee pollination and SI system, with the occurrence of late-acting self-incompatibility mechanisms in some species.