Assignment of the disulfide bridges in bothropstoxin-I, a myonecrotic Lys49 PLA(2) homolog from Bothrops jararacussu snake venom

Bothropstoxin-I (BthTX-1), a Lys49 phospholipase A(2) homolog with no apparent catalytic activity, was first isolated from Bothrops jararacussu snake venom and completely sequenced in this laboratory. It is a 121-amino-acid single polypeptide chain, highly myonecrotic, despite its inability to catalyze hydrolysis of egg yolk phospholipids, and has 14 half-cystine residues identified at positions 27, 29, 44, 45, 50, 51, 61, 84, 91, 96, 98, 105, 123, and 131 (numbering according to the conventional alignment including gaps, so that the last residue is Cys 131). In order to access its seven disulfide bridges, two strategies were followed: (1) Sequencing of isolated peptides from (tryptic + SV8) and chymotryptic digests by Edman-dansyl degradation; (2) crystallization of the protein and determination of the crystal structure so that at least two additional disulfide bridges could be identified in the final electron density map. Identification of the disulfide-containing peptides from the enzymatic digests was achieved following the disappearance of the original peptides from the HPLC profile after reduction and carboxymethylation of the digest. Following this procedure, four bridges were initially identified from the tryptic and SV8 digests: Cys50-Cys131, Cys51-Cys98, Cys61-Cys91, and Cys84-Cys96. From the chymotryptic digest other peptides were isolated either containing some of the above bridges, therefore confirming the results from the tryptic digest, or presenting a new bond between Cys27 and Cys123. The two remaining bridges were identified as Cys29-Cys45 and Cys44-Cys105 by determination of the crystal structure, showing that BthTX-1 disulfide bonds follow the normal pattern of group II PLA(2)s.

Upregulation of gp91(phox) Subunit of NAD(P)H Oxidase Contributes to Erectile Dysfunction Caused by Long-term Nitric Oxide Inhibition in Rats: Reversion by Regular Physical Training

OBJECTIVES To test the hypothesis that glyco protein 91phox (gp91(phox)) subunit of nicotinamide adenine dinucleotide phosphate [NAD(P) H] oxidase is a fundamental target for physical activity to ameliorate erectile dysfunction (ED). Vascular risk factors are reported to contribute to ED. Regular physical exercise prevents cardiovascular diseases by increasing nitric oxide (NO) production and/or decreasing NO inactivation.METHODS Male Wistar rats received the NO synthesis inhibitor N(omega)-nitro-L-arginine methyl ester (L-NAME) for 4 weeks, after which animals were submitted to a run training program for another 4 weeks. Erectile functions were evaluated by in vitro cavernosal relaxations and intracavernous pressure measurements. Expressions of gp91(phox) subunit and neuronal nitric oxidase synthase in erectile tissue, as well as superoxide dismutase activity and nitrite/nitrate (NO(x)) levels were determined.RESULTS The in vitro acetylcholine-and electrical field stimulation-induced cavernosal relaxations, as well as the increases in intracavernous pressure were markedly reduced in sedentary rats treated with L-NAME. Run training significantly restored the impaired cavernosal relaxations. No alterations in the neuronal nitric oxidase synthase protein expression (and its variant penile neuronal nitric oxidase synthase) were detected. A reduction of NO(x) levels and superoxide dismutase activity was observed in L-NAME-treated animals, which was significantly reversed by physical training. Gene expression of subunit gp91(phox) was enhanced by approximately 2-fold in erectile tissue of L-NAME-treated rats, and that was restored to basal levels by run training.CONCLUSIONS Our study shows that ED seen after long-term L-NAME treatment is associated with gp91(phox) subunit upregulation and decreased NO bioavailability. Exercise training reverses the increased oxidative stress in NO-deficient rats, ameliorating the ED. UROLOGY 75: 961-967, 2010. (C) 2009 Elsevier B.V.

If fleas developed a rite, It would be related to the dog

This article highlights a set of Wittgensteinian remarks on James Frazer's texts found in The Golden Bough, put it inside the context of creation of a conceptual framework for his renewed philosophical approach after his return to Cambridge, a period comprehended between 1929-1932, and delimits the ritualistic value of his grammatical method of philosophy.

The Noether theorem for geometric actions and area preserving diffeomorphisms on the torus

We find that within the formalism of coadjoint orbits of the infinite dimensional Lie group the Noether procedure leads, for a special class of transformations, to the constant of motion given by the fundamental group one-cocycle S. Use is made of the simplified formula giving the symplectic action in terms of S and the Maurer-Cartan one-form. The area preserving diffeomorphisms on the torus T2=S1⊗S1 constitute an algebra with central extension, given by the Floratos-Iliopoulos cocycle. We apply our general treatment based on the symplectic analysis of coadjoint orbits of Lie groups to write the symplectic action for this model and study its invariance. We find an interesting abelian symmetry structure of this non-linear problem.

PREVENCAO DE TROMBOFLEBITES PELO USO DE HEPARINA EM PACIENTES TRATADOS COM ANFOTERICINA B

The authors studied the incidence of thrombophlebitis in 41 patients treated intravenously with amphotericin B. The patients were divided in two different group: Group 1: patients treated with amphotericin B and hydrocortisone with heparin (1000 UI); Group 2: patients treated with amphotericin B and hydrocortisone. The results showed 23.81% of incidence of thrombophlebitis in Group 1 and 90% in Group 2. Thrombophlebitis in Group 1 ranged from mild to moderate without any change during the drug therapy. In Group 2, the incidence was 66.67% and the thrombophlebitis were severe being necessary the withdrawn of the drugs in 35.0% of the cases. We concluded that heparin, in low doses, in association with amphotericin B, was an efficient drug preventing or reducing the development of thrombophlebitis.

Continuous wave ultraviolet frequency upconversion due to triads of Nd3+ ions in fluoroindate glass

We have observed ultraviolet upconversion fluorescence from the 4D3/2 and 2P3/2 levels of Nd3+ in fluoroindate glass under infrared pumping. It was found that the excitation of a large population in the 4F3/2 metastable level allows to achieve strong upconversion emissions at 354 and 382 nm. A simple rate equation model reproduces the temporal behavior of the upconverted emission and allows us to estimate the energy transfer rate among three Nd3+ ions participating in the process. © 1997 American Institute of Physics.

Frequency upconversion in Nd3+-doped fluoroindate glass

We report the observation of frequency upconversion in fluoroindate glasses with the following compositions: (mol%) (39 - x)InF3-20ZnF2-20SrF2-16BaF 2-2GdF3-2NaF-1GaF3-xNdF3 (x = 0.05, 0.1, 0.5, 1, 2, 3). The excitation source was a dye laser in resonance with the 4I9/2→(2G5/2, 2G7/2) transition of the Nd3+ ions. The upconverted fluorescence spectra show emissions from ∼ 350 to ∼ 450 nm, corresponding to transitions 4D3/2→4I9/2 ;4D3/2→4I11/2; 2P3/2→ 4I9/2; 4D3/2→4I13/2; 2P3/2→4I11/2; 4D3/2→4I15/2; and 2P3/2 → 4I13/2. The dependence of the fluorescence signals on the laser intensity indicates that two laser photons participate in the process. The temporal behavior of the signal indicates that energy transfer among the Nd3+ ions is the main mechanism which contributes to upconversion at 354 and 382 nm.

Structure of cobalt (II) perchlorate adsorbed on silica gel surface chemically modified with benzimidazole molecule

Covalently attached benzimidazole molecules on silica gel surface, ≡SiL (where L = N-propyl-benzimidazole), adsorbs Co(ClO4)2 from non-aqueous solvent by forming a surface complex according to the reaction: m ≡SiL + Co(ClO4)2 → (≡SiL)mCo(ClO4)2. The equilibrium constant and the adsorption capacity, determined by applying the Langmuir equation were b = 3.0 × 103 L mol-1 and Ns= 0.098 × 10-3 mol g-1, respectively. The metal is bonded through the nitrogen atom and the perchlorate ion is not coordinated. The ESR study indicated that the complex has essentially an octahedral geometry with tetragonal distortion, with the electrons of the four nitrogen atoms interacting with the cobalt central metal ion in the equatorial plane. Only one complex species was detected on the surface.

Violet and blue light amplification in Nd3+-doped fluoroindate glasses

We report optical gain at 382 and 414 nm from Nd3+-doped fluroindate glasses after excitation with lasers operating either at 583 or 532 nm. Stimulated emission due to a frequency upconversion process results in increase of the emitted violet and blue light intensity and emission line narrowing. Large optical gain is measured by pump-probe spectroscopy using samples with various Nd3+ concentrations. © 1999 American Institute of Physics.

Study of the presence of the spores of Clostridium botulinum in honey in Brazil

The isolation of Clostridium botulinum from honey samples is described. Botulism is characterized as an intoxication provoked by ingestion of contaminated foods with this toxin. Infant botulism happens by the ingestion of spores of C. botulinum together with food that in special conditions of the intestinal tract, such as those present in babies of less than 1 year old, will allow the germination and colonization of the intestine with production and absorption of botulinic toxin. The samples were subjected to dilution and to a thermal shock and cultivated in modified CMM (Difco). Cultures were subjected to Gram smears and toxicity tests in mice. The toxic cultures were purified in RFCA (Oxoid) plates and incubated in anaerobic jars. Positive samples were typed using the mouse assay neutralization test. From the 85 honey samples analyzed, six were positive for C. botulinum (7.06%), and identified as producers of type A, B, and D toxins.

Assignment of the bisulfide bridges in bothropstoxin-I, a Myonecrotic Lys49 PLA2 homolog from bothrops jararacussu snake venom

Bothropstoxin-I (BthTX-I), a Lys49 phospholipase A2 homolog with no apparent catalytic activity, was first isolated from Bothrops jararacussu snake venom and completely sequenced in this laboratory. It is a 121-amino-acid single polypeptide chain, highly myonecrotic, despite its inability to catalyze hydrolysis of egg yolk phospholipids, and has 14 half-cystine residues identified at positions 27, 29, 44, 45, 50, 51, 61, 84, 91, 96, 98, 105, 123, and 131 (numbering according to the conventional alignment including gaps, so that the last residue is Cys 131). In order to access its seven disulfide bridges, two strategies were followed: (1) Sequencing of isolated peptides from (tryptic + SV8) and chymotryptic digests by Edman-dansyl degradation; (2) crystallization of the protein and determination of the crystal structure so that at least two additional disulfide bridges could be identified in the final electron density map. Identification of the disulfide-containing peptides from the enzymatic digests was achieved following the disappearance of the original peptides from the HPLC profile after reduction and carboxymethylation of the digest. Following this procedure, four bridges were initially identified from the tryptic and SV8 digests: Cys50-Cysl31, Cys51-Cys98, Cys61-Cys91, and Cys84-Cys96. From the chymotryptic digest other peptides were isolated either containing some of the above bridges, therefore confirming the results from the tryptic digest, or presenting a new bond between Cys27 and Cys 123. The two remaining bridges were identified as Cys29-Cys45 and Cys44-Cysl05 by determination of the crystal structure, showing that BthTX-I disulfide bonds follow the normal pattern of group II PLA2s. © 2001 Plenum Publishing Corporation.

Putative pyrophosphate phosphofructose 1-kinase genes identified in sugarcane may be getting energy from pyrophosphate

Pyrophosphate-dependent phosphofructokinase (PPi-PFK) has been detected in several types of plant cells, but the gene has not been reported in sugar cane. Using Citrux paradixi PPi-PFK gene (AF095520 and AF095521) sequences to search the sugar cane EST database, we have identified both the α and β subunits of this enzyme. The deduced amino acid sequences showed 76 and 80% similarity with the corresponding α and β subunits of C. paradisi. A high degree of similarity was also observed among the PFK β subunits when the alignment of the sugar cane sequences was compared to those of Ricinus communis and Solanum tuberosum, it appears that α and β are two distinct subunits; they were found at different concentrations in several sugar cane tissues. It remains to be determined if the different gene expression levels have some physiological importance and how they affect sucrose synthesis, export, and storage in vacuoles. A comparison between the amino acid sequences of β PFKs from a variety of organisms allowed us to identify the two critical Asp residues typical of this enzyme's activity site and the other binding sites; these residues are tightly conserved in all members of this protein family. Apparently, there are catalytic residues on the β subunit of the pyrophosphate-dependent enzyme.

Foraging activity and demographic patterns of two termite species (Isoptera: Rhinotermitidae) living in urban landscapes in southeastern Brazil

Coptotermes gestroi and Heterotermes tenuis (Isoptera: Rbinotermitidae) are important pests in southeastern Brazil causing serious economic damage. In this study we determined the demographic patterns and foraging activity of these species using mark-release-recapture and the consumption of wooden stakes. Using both the weighted mean and Lincoln index methods, population estimates ranged from ≈ 0.57 to 1.99 million individuals for C. gestroi and from ≈ 0.20 to 1.37 million for H. tenuis. Territory size of the colonies ranged from 172.5 to 5235 m 2 for C. gestroi and from 16 to 40 m 2 for H. tenuis. Our results also indicate that foraging activity was dependent on the minimum temperature; however, the existence of a compensation strategy in the foraging activities may permit foragers to exploit food sources under different environmental conditions.

Preparation of a silica gel modified with 2-amino-1,3,4-thiadiazole for adsorption of metal ions and electroanalytical application

Silica gel with a specific area of 382 m2 g-1 and an average pore diameter of 60 Å was chemically modified with 2-amino-1,3,4-thiadiazole, for the purpose of selective adsorption of heavy metals ions and possible use as a chemically modified carbon paste electrode (CMCPE). The following properties of this functionalized silica gel are discussed: selective adsorption of heavy metal ions measured by batch and chromatographic column techniques, and utilization as preconcentration agent in a chemically modified carbon paste electrode (CMCPE) for determination of mercury(II). The chemical selectivity of this functional group and the selectivity of voltammetry were combined for preconcentration and determination. ©2006 Sociedade Brasileira de Química.

Avaliação de diferentes híbridos suínos submetidos à insensibilização elétrica e gasosa (CO 2). Parte 3 - Mensurações visuais de qualidade

Three pig genetics lineages A, B and C marketed in Brazil, were stunning with the manual electric stunning (Karl Schermer 220-230/250 volts, 45-60 Hz and 1.4 - 1.5 A) and the collective gaseous system (COMBI-BUTINA 90% CO 2). The electric stunning provided higher blood splashed levels in the areas of the inside round (0.477 and 0.26, p ≤ 0.01), shoulder/cranial (0.154 and 0.039, p ≤ 0.005), shoulder/central (0.261 e 0.052, p ≤ 0.001), shoulder/caudal (0.180 and 0.030, p ≤ 0.01), loin/central (0.185 and 0.065, p ≤ 0.01), loin/caudal (0.06 and 0.207, p ≤ 0.01) and loin/lateral external (0.061 and 0.013, p ≤ 0.05), as well as more diffuse blood splashed in the areas of the inside round (0.461 and 0.279, p ≤ 0.05), shoulder/cranial (0.154 and 0.039, p ≤ 0.001), shoulder/central (0.231 and 0.039, p ≤ 0.001) and shoulder/caudal (0.185 and 0.026, p ≤ 0.001). The electric stunning also provided higher skin damage levels in the areas of the shoulder (1.098 and 0.795, p ≤ 0.001), body (1.04 and 0.948, p ≤ 0.05) and ham (0.84 and 0.68, p ≤ 0.001), as well as higher eyelid reflex levels (11.57%) comparatively to the gaseous system (2.86%) of a total of 426 pigs. Small indexes of bone fractures and muscle bruises were found in both systems.