beta(2)-Agonists and cAMP inhibit protein degradation in isolated chick (Gallus domesticus) skeletal muscle

1. The role of beta(2)-agonist and of cAMP in chick skeletal muscle proteolytic pathways and protein synthesis was investigated using an in vitro preparation that maintains tissue glycogen stores and metabolic activity for several hours.2. In extensor digitorum longus (EDL) muscle total proteolysis decreased by 15 to 20% in the presence of equimolar concentrations of epinephrine, clenbuterol, a selective beta(2)-agonist, or dibutyryl-cAMP. Rates of protein synthesis were not altered by clenbuterol or dibutyryl-cAMP.3. The decrease in the rate of total protein degradation induced by 10(-5) M clenbuterol was paralleled by a 44% reduction in Ca2+-dependent proteolysis, which was prevented by 10(-5) M ICI 118.551, a selective beta(2)-antagonist.4. No change was observed in the activity of the lysosomal, ATP-dependent, and ATP-independent proteolytic systems. Ca2+-dependent proteolytic activity was also reduced by 58% in the presence of 10(-4) M dibutyryl-cAMP or isobutylmethylxanthine.5. The data suggest that catecholamines exert an inhibitory control of Ca2+-dependent proteolysis in chick skeletal muscle, probably mediated by beta(2)-adrenoceptors, with the participation of a cAMP-dependent pathway.

EFFECTS OF ENDOTOXIN, INTERLEUKIN-1-BETA AND PROSTAGLANDIN INJECTIONS ON FEVER RESPONSE IN BROILERS

1. The effect of endotoxin, interleukin-1 beta and prostaglandin on fever response was studied in 80 broilers (Hubbard strain). Endotoxin (E. coli, LPS) was injected iv (1.5 mu g/kg) and icv (1.5 mu g/bird); interleukin-1 (human recombinant IL-1 beta, 80 pg/bird) and prostaglandin E(2) (5 mu g/bird) were injected icv. Indomethacin (10 mg/kg, iv) pretreatment was also used before iv endotoxin injection. 2. The results showed that indomethacin was able to block the fever response induced by iv endotoxin injection, and IL-1 beta and PGE(2) were both effective in producing fever when injected icv. These data suggest a prostaglandin-mediated fever response by broilers, and also a strong evidence of the involvement of endogenous pyrogen (interleukin-1) in fever response in birds.

New crystal forms of Diocleinae lectins in the presence of different dimannosides

Studying the interactions between lectins and sugars is important in order to explain the differences observed in the biological activities presented by the highly similar proteins of the Diocleinae subtribe. Here, the crystallization and preliminary X--ray data of Canavalia gladiata lectin (CGL) and C. maritima lectin (CML) complexed with Man(alpha 1-2)Man(alpha 1)OMe, Man(alpha 1-3)Man(alpha 1)OMe and Man(alpha 1-4)Man(alpha 1)OMe in two crystal forms [the complexes with Man(alpha 1-3)Man(alpha 1)OMe and Man(alpha 1-4)Man(alpha 1)OMe crystallized in space group P3(2) and those with Man(alpha 1-2)Man(alpha 1)OMe crystallized in space group I222], which differed from those of the native proteins (P2(1)2(1)2 for CML and C222 for CGL), are reported. The crystal complexes of ConA-like lectins with Man(alpha 1-4)Man(alpha 1)OMe are reported here for the first time.

RELATIONSHIP BETWEEN PLASMA 17-BETA ESTRADIOL ON THE DAY OF ESTRUS AND NUMBER OF VIABLE EMBRYOS IN BOS-INDICUS (NELLORE) COWS SUPEROVULATED WITH FSH

Thirteen cows, Bos indicus, of the Nellore breed were superovulated with 22 mg of follicle stimulating hormone (FSH) administered by intramuscular route during four consecutive days (D10, D11, D12 and D13), starting on the 10th day of the estrous cycle (day 0 = estrus). Prostaglandin (PGF2alpha, 1.0 mg, im) was administered on D12, 48 h after the first FSH injection, for the induction of estrus on D14, when artificial insemination was performed. Seven days later (D21 of the cycle), embryos were collected, and evaluated, and the ovarian response was estimated on the basis of number of corpora lutea determined by rectal palpation. Blood samples were obtained for the determination of plasma 17-beta estradiol on D10, D11, D12, D13, D14 and D21 and plasma progesterone on D14 by RIA. The donors were divided into two groups according to progesterone levels on D14, the day of the induced estrus (GI: P4 less-than-or-equal-to 1.00 ng/ml, N = 5 and GII: P4 > 1.00 ng/ml, N = 8). A linear positive correlation was observed between plasma 17-beta estradiol concentration on the day of estrus and viable embryo number. We conclude that plasma 17-beta estradiol and progesterone concentrations on the day of estrus can be used to predict the viability of embryos recovered from Nellore cows superovulated with FSH.

Crystallization and preliminary X-ray diffraction analysis of prephenate dehydratase from Mycobacterium tuberculosis H37Rv

Tuberculosis remains the leading cause of mortality arising from a bacterial pathogen ( Mycobacterium tuberculosis). There is an urgent need for the development of new antimycobacterial agents. The aromatic amino-acid pathway is essential for the survival of this pathogen and represents a target for structure-based drug design. Accordingly, the M. tuberculosis prephenate dehydratase has been cloned, expressed, purified and crystallized by the hanging-drop vapour-diffusion method using PEG 400 as a precipitant. The crystal belongs to the orthorhombic space group I222 or I2(1)2(1)2(1), with unit-cell parameters a = 98.26, b = 133.22, c = 225.01 angstrom, and contains four molecules in the asymmetric unit. A complete data set was collected to 3.2 angstrom resolution using a synchrotron-radiation source.

Crystallization and preliminary X-ray diffraction analysis of XAC1151, a small heat-shock protein from Xanthomonas axonopodis pv. citri belonging to the alpha-crystallin family

The hspA gene (XAC1151) from Xanthomonas axonopodis pv. citri encodes a protein of 158 amino acids that belongs to the small heat-shock protein ( sHSP) family of proteins. These proteins function as molecular chaperones by preventing protein aggregation. The protein was crystallized using the sitting-drop vapour-diffusion method in the presence of ammonium phosphate. X-ray diffraction data were collected to 1.65 angstrom resolution using a synchrotron-radiation source. The crystal belongs to the rhombohedral space group R3, with unit-cell parameters a = b = 128.7, c = 55.3 angstrom. The crystal structure was solved by molecular-replacement methods. Structure refinement is in progress.

Effect of acute heat stress on heat shock protein 70 messenger RNA and on heat shock protein expression in the liver of broilers

1. The synthesis of heat shock protein 70 (Hsp70) mRNA and the expression of Hsp70 in the liver of broiler chickens submitted to acute heat stress (35 degrees C for 5 h) was investigated.2. Hsp70 expression was detected by SDS-PAGE and Western blot analysis using a polyclonal antiserum against Hsp70 of Blastocladiella emersonii. The specific signal of Hsp70 mRNA was analysed by Northern blot using as probe a Hsp70 cDNA of B. emersonii.3. An increase in the amount of Hsp70 was detected from the first up to the fifth hour of acute heat exposure. This increase in the amount of Hsp70 was accompanied by an increase in Hsp70 mRNA which peaked at 3 h.4. This study shows that the heat induced increase in Hsp70 mRNA and protein in broiler liver, in vivo, are time dependent, similar to that in mammals.

Reproductive modes of Atlantic forest hylid frogs: A general overview and the description of a new mode

This paper reports a new reproductive mode in anurans, observed for the green treefrog, Hyla leucopygia, a species from the Atlantic Forest of Brazil and a member of the H. albosignata group. The reproductive mode of H. leucopygia is defined as follows: aquatic eggs and embryos in subterranean constructed nests; subsequent to flooding, feeding tadpoles in ponds or streams. We suggest that the new reproductive mode is widespread among the species in the H. albosignata group. The specialized courtship behaviors of H. leucopygia, characterized by vocalizations and a stereotyped sequence of mutual touches between male and female, are described. A review of the reproductive modes of hylid frogs is provided, with 11 modes recognized for the Atlantic Forest hylids and 12 modes For hylids around the world.