Acanthoscurrin Fragment 101-132: Total Synthesis at 60 degrees C of a Novel Difficult Sequence

Glycine-rich proteins (GRP), serve a variety of biological functions. Acanthoscurrin is an antimicrobial GRP isolated front hemocytes-of the Brazilian spider Acanthoscurria gomesiana. Aiming to contribute to the knowledge of the secondary structure and stepwise solid-phase synthesis of GRPs` glycine-rich domains, we attempted to prepare G(101)GGLGGGRGGGYG(113) GGGGYGGGYG(123)GGy(126)GGGKYK(132)-NH(2), acanthoscurrin C-terminal amidated fragment. Although a theoretical prediction did not indicate high aggregation potential for this peptide, repetitive incomplete aminoacylations were observed after incorporating Tyr(126) to the growing peptide-MBHA resin (Boc chemistry) at 60 degrees C. The problem was not solved by varying the coupling reagents or solvents, adding chaotropic salts to the reaction media or changing the resin/chemistry (Rink amide resin/Fmoc chemistry). Some improvement was mode when CLEAR amide resin (Fmoc chemistry) was 32 used, as it allowed for obtaining fragment (G(113)-K(132) NIR-FT-Raman spectra collected for samples of the growing peptide-MBHA, -Rink amide resin and -CLEAR amide resin revealed the presence of beta-sheet structures. Only the combination of CLEAR-amide resin, 60 degrees C, Fmoc-(Fmoc-Hmb)Gly-OH and LiCl (the last two used alternately) was able to inhibit the phenomenon, as proven by NIR-FT-Raman analysis of the growing peptide-resin, allowing the total synthesis of desired 132 fragment Gly(101)-K(132). In summary, this work describes a new difficult sequence, contributes to understanding stepwise solid-phase synthesis of this type of peptide and shows that, at least while protected and linked to a resin, this GRPs glycine-rich motif presents all early tendency to assume beta-sheet structures. (c) 2008 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 92: 65-75, 2009.

Microwave-assisted solid-phase peptide synthesis at 60 degrees C: alternative conditions with low enantiomerization

Several conditions have been used in the coupling reaction of stepwise SPPS at elevated temperature (SPPS-ET), but we have elected the following as our first choice: 2.5-fold molar excess of 0.04-0.08 M Boc or Fmoc-amino acid derivative, equimolar amount of DIC/HOBt (1:1)or TBTU/DIPEA(1:3), 25% DMSO/toluene, 60 degrees C, conventional heating. In this study, aimed to further examine enantiomerization under such condition and study the applicability of our protocols to microwave-SPPS, peptides containing L-Ser, L-His, L-Cys and/or L-Met were manually synthesized traditionally, at 60 degrees C using conventional heating and at 60 degrees C using microwave heating. Detailed assessment of all crude peptides (in their intact and/or fully hydrolyzed forms) revealed that, except for the microwave-assisted coupling of L-Cys, all other reactions occurred with low levels of amino acid enantiomerization (<2%). Therefore, herein we (i) provide new evidences that our protocols for SPPS at 60 degrees C using conventional heating are suitable for routine use, (ii) demonstrate their appropriateness for microwave-assisted SPPS by Boc and Fmoc chemistries, (iii) disclose advantages and limitations of the three synthetic approaches employed. Thus, this study complements our past research on SPPS-ET and suggests alternative conditions for microwave-assisted SPPS. Copyright (C) 2009 European Peptide Society and John Wiley & Sons, Ltd.

Decay resistance of thermally-modified Eucalyptus grandis wood at 140 degrees C, 160 degrees C, 180 degrees C, 200 degrees C and 220 degrees C

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Modeling a system of phosphated cross-linked high amylose for controlled drug release. Part 2: Physical parameters, cross-linking degrees and drug delivery relationships

High amylose cross-linked to different degrees with sodium trimetaphosphate by varying base strength (2% or 4%) and contact time (0.5-4 h) was evaluated as non-compacted systems for sodium diclophenac controlled release. The physical properties and the performance of these products for sodium diclophenac controlled release from non-compacted systems were related to the structures generated at each cross-linking degree. For samples at 2% until 2 h the swelling ability, G' and eta* values increased with the cross-linking degree, because the longer polymer chains became progressively more entangled and linked. This increases water uptake and holding, favoring the swelling and resulting in systems with higher viscosities. Additionally, the increase of cross-linking degree should contribute for a more elastic structure. The shorter chains with more inter-linkages formed at higher cross-linking degrees (2%4h and 4%) make water caption and holding difficult, decreasing the swelling, viscosity and elasticity. For 2% samples, the longer drug release time exhibited for 2%4h sample indicates that the increase of swelling and viscosity contribute for a more sustained drug release, but the mesh size of the polymeric network seems to be determinant for the attachment of drug molecules. For the 4% samples, smaller meshes size should determine less sustained release of drug. (C) 2008 Elsevier B.V. All rights reserved.

Functional alterations in endocrine pancreas of rats with different degrees of dexamethasone-induced insulin resistance

Objectives: We have analyzed the peripheral insulin and glucose sensitivity in vivo, and islet function ex vivo in rats with different degrees of insulin resistance induced by dexamethasone (DEX).Methods: Dexamethasone, in the concentrations of 0.1 (DEX 0.1), 0.5 (DEX 0.5), and 1.0 mg/kg body weight (DEX 1.0) was administered daily, intraperitoneally, to adult Wistar rats for 5 days, whereas controls received saline.Results: Dexamethasone treatment induced peripheral insulin resistance in a dose-dependent manner. At the end of the treatment, only DEX 1.0 rats showed significant increase of postabsorptive blood glucose and serum triglycerides, and nonesterified fatty acids levels. Incubation of pancreatic islets in increasing glucose concentrations (2.8-22 mM) led to an augmented insulin secretion in all DEX-treated rats. Leucine, carbachol, and high KCl concentrations induced the insulin release in DEX 0.5 and DEX 1.0, whereas arginine augmented secretion in all DEX-treated groups.Conclusions: We demonstrate that in DEX 0.5 and, especially in DEX 0.1 groups, but not in DEX 1.0, the adaptations that occurred in the endocrine pancreas are able to counteract metabolic disorders (glucose intolerance and dyslipidemia). These animal models seem to be interesting approaches for the study of degrees of subjacent effects that may mediate type 2 diabetes (DEX 1.0) and islet function alterations, without collateral effects (DEX 0.1 and DEX 0.5).

Reproduction of the speckled swimming crab Arenaeus cribrarius (Brachyura : portunidae) on the Brazilian coast near 23 degrees 30 ' S

The reproductive biology of Arenaeus cribrarius from Ubatuba, São Paulo State, Brazil, was studied. Swimming crabs were sampled monthly for two years with otter trawls in two bays. A total of 941 males and 1,012 females were examined. Mating took place mainly in autumn involving postmolt females and intermolt males. At that time, gonad regression was verified in adult males, due to spermatophore transfer, and the molting of adult females. Ovigerous females or females with mature gonads were present year-round but more frequently captured during spring and summer. We found that 19, of all adult females were premolt, which indicated the occurrence of another mature instar and thus the absence of a well-defined terminal molt after puberty. Intermolt males were captured throughout the whole study period.

Population structure, distribution and abundance patterns of the patagonian smoothhound Musmlus schmitti springer, 1939 (Chondrichthyes,Elasmobranchii, Triakidae) in the rio de la plata and inner continental shelf, SW Atlantic ocean (34 degrees-30'-39 degrees-30's)

A total number of 4824 Mustelus schmitti was sampled. Females ranged from 25 to 93 em in spring and from 28 to 90 cm in summer. Males ranged from 34 to 82 and from 28 to 77 cm, respectively. Length composition of the population showed significant differences between spring and summer being females larger than males. Total length distribution did not show significant differences between cruises. Males density varied significantly between cruises while for the females no significant variation was observed. In the spring cruise, both sexes occurred at depths lower than 50 in. Females occurred in the whole area with adult occurrence only above 35 degrees 30'S. Mature males occurred throughout the area, immature males occurring in two trawls in Samborombon Bay. The summer cruise showed a discontinuous distribution of the species along the study area resulting in spatial segregation in two groups with immature females predominating in both of them.

Size at maturity and egg capsules of the softnose skates Bathyraja brachyurops (Fowler, 1910) and Bathyraja macloviana (Norman, 1937) (Elasmobranchii: Rajidae) in the SW Atlantic (37 degrees 00'-39 degrees 30'S)

P>The softnose skates Bathyraja brachyurops and Bathyraja macloviana represent an important portion of the skate catches of the Uruguayan trawling fleet in the southwestern Atlantic. From March to October 2004, specimens of these species were collected at 75-200 m depth range in the area situated between latitudes 37 degrees 00'-39 degrees 30'S. For B. brachyurops, total length at which 50% of the specimens were retained by the gear was 68.0 cm for both sexes; T(L50) was estimated at 65.4 cm for males and 67.0 cm for females. For B. macloviana, total length at which 50% of the specimens were retained was 56.0-57.0 cm for both sexes; T(L50) was estimated at 53.5 cm for males and 52.0 cm for females. Egg capsule length varied from 79-91 mm in B. brachyurops and 69-75.5 mm in B. macloviana. In both species, capsules displayed striated surfaces and similar gross morphology, although egg capsules of B. macloviana had more robust anterior horns and a smaller size than those of B. brachyurops. Egg capsules of the latter also exhibited microscopical prickles. Capsule edges were laterally keeled with a groove along the keel, and a straight and transverse velum was present in the egg capsules of both species.

Seasonal changes in daily metabolic patterns of tegu lizards (Tupinambis merianae) placed in the cold (17 degrees C) and dark

Oxygen consumption rate was measured continuously in young tegu lizards Tupinambis merianae exposed to 4 d at 25 degrees C followed by 7-10 d at 17 degrees C in constant dark at five different times of the year. Under these conditions, circadian rhythms in the rate of oxygen consumption persisted for anywhere from 1 d to the entire 2 wk in different individuals in all seasons except the winter. We also saw a progressive decline in standard oxygen consumption rate (at highly variable rates in different individuals) to a very low rate that was seasonally independent (ranging from 19.1 +/- 6.2 to 27.7 +/- 0.2 mL kg(-1) h(-1) across seasons). Although this degree of reduction appeared to take longer to invoke when starting from higher metabolic rates, tegu lizards reduced their metabolism to the low rates seen in winter dormancy at all times of the year when given sufficient time in the cold and dark. In the spring and summer, tegus reduced their standard metabolic rate (SMR) by 80%-90% over the experimental run, but only roughly 20%-30% of the total fall was due to the reduction in temperature; 70%-80% of the total fall occurred at constant temperature. By autumn, when the starting SMR on the first night at 25 degrees C was already reduced by 59%-81% (early and late autumn, respectively) from peak summer values, virtually all of the fall (63%-83%) in metabolism was due to the reduction in temperature. This suggests that the temperature-independent reduction of metabolism was already in place by autumn before the tegus had entered winter dormancy.

Infrared degrees of freedom of Yang-Mills theory in the Schrodinger representation

We set up a new calculational framework for the Yang-Mills vacuum transition amplitude in the Schrodinger representation. After integrating out hard-mode contributions perturbatively and performing a gauge-invariant gradient expansion of the ensuing soft-mode action, a manageable saddle-point expansion for the vacuum overlap can be formulated. In combination with the squeezed approximation to the vacuum wave functional this allows for an essentially analytical treatment of physical amplitudes. Moreover, it leads to the identification of dominant and gauge-invariant classes of gauge field orbits which play the role of gluonic infrared (IR) degrees of freedom. The latter emerge as a diverse set of saddle-point solutions and are represented by unitary matrix fields. We discuss their scale stability, the associated virial theorem and other general properties including topological quantum numbers and action bounds. We then find important saddle-point solutions (most of them solitons) explicitly and examine their physical impact. While some are related to tunneling solutions of the classical Yang-Mills equation, i.e. to instantons and merons, others appear to play unprecedented roles. A remarkable new class of IR degrees of freedom consists of Faddeev-Niemi type link and knot solutions, potentially related to glueballs.