Binding affinities and activation of Asp712Ala and Cys100Ser mutated kinin B1 receptor forms suggest a bimodal scheme for the molecule of bound-DABK

AbstractMutant forms of kinin B1 receptor (B1R) and analogs of the full agonist des-Arg9-bradykinin (DABK) were investigated aiming to verify the importance of selected receptor residues and of each agonist-peptide residue in the specific binding and activation. Linked by a specific disulfide bond (Cys100–Cys650), the N-terminal (Nt) and the EC3 loop C-terminal (Ct) segments of angiotensin II (AngII) receptor 1 (AT1R) have been identified to form an extracellular site for binding the agonist Nt segment (Asp1 and Arg2 residues). Asp712 residue at the receptor EC3 loop binds the peptide Arg2 residue. By homology, a similar site might be considered for DABK binding to B1R since this receptor contains the same structural elements for composing the site in AT1R, namely the disulfide bond and the EC3 loop Asp712 residue. DABK, Alan-DABK analogs (n=Ala1-, Ala2-, Ala3-, Ala4-, Ala5-, Ala6-, Ala7-, Ala8-DABK), and other analogs were selected to binding wild-type, Asp712Ala and Cys100Ser mutated B1R receptors. The results obtained suggested that the same bimodal scheme adopted for AngII-AT1R system may be applied to DABK binding to B1R. The most crucial similarity in the two cases is that the Nt segments of peptides equally bind to the homologous Asp712 residue of both AT1R and B1R extracellular sites. Confirming this preliminary supposition, mutation of residues located at the B1R extracellular site as EC3 loop Asp712 and Cys100 caused the same modifications in biological assays observed in AT1R submitted to homologous mutations, such as significant weakening of agonist binding and reduction of post-receptor-activation processes. These findings provided enough support for defining a site that determines the specific binding of DABK to B1R receptors

Sexing freshwater turtles: penile eversion in Phrynops tuberosus (Testudines: Chelidae)

Here, we described a noninvasive method for sexing freshwater turtles by stimulating penile eversion. We immobilized the neck and limbs of animals using fingers and, after some seconds, turtles everted their penis. This method was tested in 33 male Phrynops tuberosus, and 28 everted the penis. The efficiency of the method was not dependent of animal size, which reinforces its applicability. Our method allows sexing turtles in the field, avoiding killing the animal or causing major injuries in order to assess the sex

ER stress activation in the intestinal mucosa but not in mesenteric adipose tissue is associated with inflammation in Crohn’s disease patients

Chronic/abnormal activation of endoplasmic reticulum (ER) stress is linked to the exacerbation of the inflammatory process and has been recently linked to Crohn’s disease (CD) pathophysiology. We investigated the intestinal mucosa and the mesenteric adipose tissue (MAT) collected from CD patients with active disease (CD group) and from non-IBD patients (CTR group) to study ER stress activation and to address tissue-specific modulation in CD. The intestinal mucosa of CD patients showed an upregulation in the expression of ER stress related genes, including ATF3, DNAJC3, STC2, DDIT3, CALR, HSPA5 and HSP90B1. Results showed that EIF2AK3 gene was upregulated, along with increased protein expression of p-eIF2α and p-eIF2α/eIF2α ratio. Additionally, ERN1 gene expression was upregulated, along with an increased spliced/activated form sXBP1 protein. Despite the upregulation of ATF6 gene expression in the intestinal mucosa of CD patients, no differences were found in ATF6 protein expression. Lastly, the analysis of MAT revealed unchanged levels of ER stress markers along with no differences in the activation of UPR. However, chaperone gene expression was modulated in the MAT of CD patients. To conclude, our results address tissue-specific differences in UPR activation in CD and point the ER stress as an important pro-inflammatory mechanism in CD, specifically in the intestinal mucosa149CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP401270/2016-502-P-28707/20122016/01638-

PROFIT: a new alternative for emission-line PROfile FITting

I briefly describe a simple routine for emission-line profiles fitting by Gaussian curves or Gauss-Hermite series. The PROFIT (line-PROfile FITting) routine represent a new alternative for use in fits data cubes, as those from Integral Field Spectroscopy or Fabry-Perot Interferometry, and may be useful to better study the emission-line flux distributions and gas kinematics in distinct astrophysical objects, such as the central regions of galaxies and star forming regions. The PROFIT routine is written in IDL language and is available at http://www.ufsm.br/rogemar/software.html. The PROFIT routine was used to fit the [Fe II]1.257um emission-line profiles for about 1800 spectra of the inner 350 pc of the Seyfert galaxy Mrk1066 obtained with Gemini NIFS and shows that the line profiles are better reproduced by Gauss-Hermite series than by the commonly used Gaussian curves. The two-dimensional map of the h_3 Gauss-Hermite moment shows its highest absolute values in regions close to the edge of the radio structure. These high values may be originated in an biconical outflowing gas associated with the radio jet - previously observed in the optical [O III] emission. The analysis of this kinematic component indicates that the radio jet leaves the center of the galaxy with the north-west side slightly oriented towards us and the south-east side away from us, being partially hidden by the disc of the galaxy.Comment: Accepted for publication Astrophysics & Space Science - 7 pges; 4 Fig

Review: Heads or Tails? Toward a Clear Role of Biochar as a Feed Additive on Ruminant’s Methanogenesis

The use of biochar has been suggested as a promising strategy in bio-waste management and greenhouse gases mitigation. Additionally, its use, as a feed additive, in ruminants has been reported to have contrasting effects on enteric methane production. Hence, this chapter intends to overview the most relevant literature that exploited the use of biochar as a mitigation strategy for methane. This includes the reported effects of biochar on methane production and rumen fermentation observed in in vitro and in vivo assays, as well as manure’s methane emission. The information available about the biochar and the experimental conditions used in the different studies is still limited, which created additional challenges in identifying the biological mechanisms that potentially drive the contrasting results obtained. Nevertheless, it is clear from the current state-of-the-art that biochar may be a key player in the modulation of gut fermentation and in the reduction of greenhouse gases produced by ruminants that need to be consolidated by further research