SP-Sephadex equilibrium chromatography of bradykinin and related peptides: Application to trypsin-treated human plasma

An analytical method is deseribed for the separation of bradykinin, Lys-bradykinin, and Met-Lys-bradykinin by equilibrium chromatography on SP-Sephadex C-25 eluted in 0.02 Tris-HCl buffer, pH 8.10, 0.12 NaCl. A second elution buffer, 0.02 Tris-HCl buffer, pH 7.70, 0.06 NaCl, serves as a second parameter for the identification of bradykinin and also separates the hormone from plasma bradykinin-potentiating peptides. Ten to one-hundred nanomoles of each peptide can be recovered in high yields, identified by elution position, and measured by bioassay with the isolated guinea pig ileum. The identification of bradykinin as the peptide released by trypsin acting on acid-denatured plasma is documented as an illustration of the method

Beachrocks of the northeast of Brazil: local effects of sea level fluctuations in a far-field during in Holocene

Along the coast of the state of Pernambuco, NE Brazil, the presence of coastal outcrops aligned with the shoreline and distributed above and below the mean sea level is remarkable. These outcrops were sampled, and petrological, isotopic, and geochronological analyses were integrated to investigate the local effects of the Holocene sea-level fluctuations. The results indicate that the rocks are recent (< 8ka), formed by cementation of beach sands (magnesian calcite cement, predominance of quartz and presence of marine bioclasts, signs of compaction, fractures, and dissolution), in a tropical climate and warm waters, which allows to classify them as typical examples of beachrocks. The carbonate cement consisted of Mg-rich calcite with values ranging from −1.1‰ to 3.5‰ for δ13C and from −0.9‰ to 0.5‰ for δ18OVPDB. These isotopic values are typical of marine carbonate cemented deposits in shallow marine environments under freshwater influence (meteoric vadose environment). In these conditions, the cementing processes occurred in the intertidal zones, which reinforce the use of these data as indicators of ancient sea levels. The sea level fluctuations could be divided, in chronological order, into three different phases, starting with a rapid sea level rise followed by a relative stabilization at a maximum level and, finally, a decreasing phase. Recent sea level fluctuations in Pernambuco are represented by beachrocks with ages between 8 ka and 800 years and a maximum level of 2 to 3 meters above the present mean level. In scenarios of a near future with a general global sea level rise pattern, significant local geomorphological changes may occur, which means a great future challenge to society

A structure-based site-directed mutagenesis study on the neurolysin (EC 3.4.24.16) and thimet oligopeptidase (EC 3.4.24.15) catalysis

Neurolysin (EP24.16) and thimet oligopeptidase (EP24.15) are closely related metalloendopeptidases. Site-directed mutagenesis of Tyr(613) (EP24.16) or Tyr(612) (EP24.15) to either Phe or Ala promoted a strong reduction of k(cat)/K-M for both enzymes. These data suggest the importance of both hydroxyl group and aromatic ring at this specific position during substrate hydrolysis by these peptidases. Furthermore, the EP24.15 A607G mutant showed a k(cat)/K-M of 2x10(5) M-1 s(-1) for the Abz-GFSIFRQ-EDDnp substrate, similar to that of EP24.16 (k(cat)/K-M = 3x10(5) M-1 s(-1)) which contains Gly at the corresponding position; the wild type EP24.15 has a k(cat)/K-M of 2.5x10(4) M-1 s(-1) for this substrate. (C) 2003 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.Univ Mogi das Cruzes, CIIB, BR-08780911 Mogi Das Cruzes, SP, BrazilInst Butantan, CAT, Ctr Toxicol Aplicada, BR-05467010 São Paulo, BrazilUniv São Paulo, Inst Ciencias Biomed, Program Biol Celular, Dept Histol & Embriol, BR-05508900 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Biofis, BR-04044020 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Biofis, BR-04044020 São Paulo, BrazilWeb of Scienc

Physicochemical Properties of Sugarcane Industry Residues Aiming at Their Use in Energy Processes

According to the US Department of Agriculture, sugarcane global production for marketing year (MY) 2020/21 will forecast up 22 million tons in comparison with 2019/2020 MY, reaching 188 million tons (raw value), due to higher production in Brazil, India and Thailand. These countries alternate sugarcane uses for obtaining sugar, ethanol and other products, generating near to 152 million tons of residues. In a circular economy context, the reuse of the sugarcane industry by-products is desired. Nowadays, bagasse and, sometimes, straw are used for energy recovery through combustion, while filter cake and vinasse are commonly used for soil fertilization. However, while bagasse and straw present potential for energy recovery through the thermochemical route, vinasse and filter cake are better applied in anaerobic digestion processes to produce biogas and biofertilizer. These treatments, when correctly employed, can improve the performance of sugarcane industry by diversifying its energy sources and products. For this, the correct design of equipment and processes is essential, which requires the knowledge of physical and chemical properties of sugarcane industry’s by-products. In this context, the chapter goal is to present an updated literature review for these properties, considering their use in energy recovery processes

Activation of leukocyte rolling by the cysteine-rich domain and the hyper-variable region of HF3, a snake venom hemorrhagic metalloproteinase

AbstractThe functionality of the disintegrin-like/cysteine-rich domains of snake venom metalloproteinases (SVMPs) has been shown to reside in the cysteine-rich region, which can interact with VWA-containing proteins. Recently, the hyper-variable region (HVR) of the cysteine-rich domain was suggested to constitute a potential protein–protein adhesive interface. Here we show that recombinant proteins of HF3, a hemorrhagic P-III SVMP, containing the cysteine-rich domain (disintegrin-like/cysteine-rich and cysteine-rich proteins) but not the disintegrin-like protein were able to significantly increase leukocyte rolling in the microcirculation. Peptides from the HVR also promoted leukocyte rolling and this activity was inhibited by anti-alphaM/beta2 antibodies. These results show, for the first time, that the cysteine-rich domain and its HVR play a role in triggering pro-inflammatory effects mediated by integrins

Structural features that make oligopeptides susceptible substrates for hydrolysis by recombinant thimet oligopeptidase

A systematic analysis of the peptide sequences and lengths of several homologues of bioactive peptides and of a number of quenched-fluorescence (qf) opioid- and bradykinin-related peptides was performed to determine the main features leading the oligopeptides to hydrolysis by the recombinant rat testis thimet oligopeptidase (EC 3.4.24.15). The results indicate that a minimum substrate length of six amino acids is required and that among the oligopeptides six to thirteen amino acid residues long, their susceptibility as substrates is highly variable. Thimet oligopeptidase was able to hydrolyse, with similar catalytic efficiency, peptide bonds having hydrophobic or hydrophilic amino acids as well as proline in the P1 position of peptides, ranging from a minimum of six to a maximum of approximately thirteen amino acid residues. An intriguing observation was the shift of the cleavage site, at a Leu-Arg bond in qf dynorphin (2-8) [qf-Dyn(2-8); Abz-GGFLRRV-EDDnp, where Abz stands for o-aminobenzoyl and EDDnp for N-(2,4-dinitrophenyl) ethylenediamine], to Arg-Arg in qf-Dyn(2-8)Q, in which Gln was substituted for Val at its C-terminus. Similarly, a cleavage site displacement was also observed with the hydrolysis of the internally quenched-fluorescence bradykinin analogues containing Gin at the C-terminal position, namely Abz-RPPGFSPFR-EDDnp and Abz-GFSPFR-EDDnp are cleaved at the Phe-Ser bond, but Abz-RPPGFSPFRQ-EDDnp and Abz-GFSPFRQ-EDDnp are cleaved at the Pro-Phe bond.UNIV FED SAO PAULO,ESCOLA PAULISTA MED,DEPT BIOFIS,BR-04044020 SAO PAULO,SP,BRAZILINST BUTANTAN,BIOCHEM & BIOPHYS LAB,BR-05503900 SAO PAULO,BRAZILUNIV SAO PAULO,INST BIOMED SCI,SAO PAULO,BRAZILFDN ANTONIO PRUDENTE,BR-01509010 SAO PAULO,BRAZILUNIV FED SAO PAULO,ESCOLA PAULISTA MED,DEPT BIOFIS,BR-04044020 SAO PAULO,SP,BRAZILWeb of Scienc

INSECTICIDE-TREATED BED NETS IN RONDÔNIA, BRAZIL: EVALUATION OF THEIR IMPACT ON MALARIA CONTROL

Mosquito nets treated with long-lasting insecticide (LLINs), when used in compliance with guidelines of the World Health Organization, may be effective for malaria vector control. In 2012, approximately 150,000 LLINs were installed in nine municipalities in the state of Rondônia. However, no studies have assessed their impact on the reduction of malaria incidence. This study analyzed secondary data of malaria incidence, in order to assess the impact of LLINs on the annual parasite incidence (API). The results showed no statistically significant differences in API one year after LLIN installation when compared to municipalities without LLINs. The adoption of measures for malaria vector control should be associated with epidemiological studies and evaluations of their use and efficiency, with the aim of offering convincing advantages that justify their implementation and limit malaria infection in the Amazon Region