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Secretory ribonucleases in the primitive ruminant chevrotain (Tragulus javanicus).

Phylogenetic analyses of secretory ribonucleases or RNases 1 have shown that gene duplication events, giving rise to three paralogous genes (pancreatic, seminal and brain RNase), occurred during the evolution of ancestral ruminants. A higher number of paralogous sequences are present in chevrotain (Tragulus javanicus), the earliest diverged taxon within the ruminants. Two pancreatic RNase sequences were identified, one encoding the pancreatic enzyme, the other encoding a pseudogene. The identity of the pancreatic enzyme was confirmed by isolation of the protein and N-terminal sequence analysis. It is the most acidic pancreatic ribonuclease identified so far. Formation of the mature enzyme requires cleavage by signal peptidase of a peptide bond between two glutamic acid residues. The seminal-type RNase gene shows features of a pseudogene, like orthologous genes in other ruminants investigated with the exception of the bovine species. The brain-type RNase gene of chevrotain is expressed in brain tissue. A hybrid gene with a pancreatic-type N-terminal and a brain-type C-terminal sequence has been identified but nothing is known about its expression. Phylogenetic analysis of RNase 1 sequences of six ruminant, three other artiodactyl and two whale species support previous findings that two gene duplications occurred in a ruminant ancestor. Three distinct groups of pancreatic, seminal-type and brain-type RNases have been identified and within each group the chevrotain sequence it the first to diverge. In taxa with duplications of the RNase gene (ruminants and camels) the gene evolved at twice as fast than in taxa in which only one gene could be demonstrated; in ruminants there was an approximately fourfold increase directly after the duplications and then a slowing in evolutionary rate.

Animals↗

Arteries of the hindfoot of the llama (Lama glama).

The objective of this study was to describe the arterial distribution of the hindfoot of the llama (Lama glama). Ten adult llamas, preserved in 6% formalin solution at 0 degree C, were dissected. The arterial system was perfused with a solution of 14% coloured plaster; the venous system was perfused with a solution of 17% coloured industrial gelatin. Angiographies were also obtained. In the llama, the arterial distribution is of the saphenous type and this simple sort of irrigation could be used as a didactic model. The caudal branch of the saphenous artery divides into the small lateral plantar artery and the larger medial plantar artery, which continues as the plantar common digital artery III, and it is the main blood supply of the hindfoot. The dorsal pedal artery is underdeveloped and the perforating tarsal artery does not exist in this species. The plantar common digital artery III divides into the plantar proper digital II, III and IV. Branches from the plantar proper digital artery III supply the digits. We compared the arterial distribution of the hindfoot of the llama with that of other domestic animals including the one-humped camel (Camelus dromedarius).

Angiography↗

Urinary retention in two male dromedaries due to silica uroliths.

Two cases of urinary retention in male dromedaries due to silica uroliths are presented. The animals were urethrostomized and one dromedary was euthanized 7 months later due to urethrostomy obstruction. The estimated daily intake of silica was 84.44 g. Silica, calcium, phosphorus and pH in urine were determined in healthy animals from the same farms, and the following values were obtained: 174.78 +/- 3.46 mg/l (silica), 41.48 +/- 4.82 mg/l (calcium), 20.4 +/- 3.6 mg/l (phosphorus) and pH 8.2, respectively. Possible causes of urolithiasis were the early castration of the animals and the fact that salt was not added to the diet. To our knowledge, this is the first description of silica urolithiasis in the dromedary camel.

Animal Feed↗

Characteristics of ovarian follicle development in domestic animals.

In most domestic animals the later stages of follicle development occurs in a wave-like pattern during oestrous cycles (cattle, sheep, goats, horses and buffalo) or periods of reproductive activity (llamas and camels). A follicle wave is the organized development of a cohort of gonadotrophin-dependent follicles all of which initially increase in size, but most of which subsequently regress and die by atresia (subordinate follicles). The number of remaining (dominant) follicles is specific to the species and is indicative of litter size. Follicle waves develop during both luteal and follicular phases and it is the dominant follicle(s) of the last follicular wave that ovulates. However, there are cases where dominant follicles from the last two follicle waves can ovulate (sheep and goats). There are exceptions to the organized wave-like pattern of follicle growth where follicle development is apparently continuous (pigs and chickens). In these animals many follicles develop to intermediate diameters and at specific times follicles that are destined to ovulate are selected from this pool and continue growing to ovulation. Understanding the pattern of follicle development in different species is increasingly important for designing improved methods to manipulate reproduction in domestic animals.

Animals↗

Electrocardiograms from the turtle to the elephant that illustrate interesting physiological phenomena.

This article describes a collection of ECGs from many species obtained over the past 50 years. Presented are ECGs of species in which the pacemaker is a separate contractile chamber with its own action and recovery potentials. In such species, pacemaker atrial and AV block can be produced. Shortening of the atrial refractory period and the negative inotropic effect can be produced by vagal stimulation. The cardiac electrogram and stroke volume are recorded from the turtle heart. The ECG and respiration were recorded from the snake. ECG records were obtained from the anesthetized and decapitated housefly. ECG records of the rabbit show slowing when the nose encountered irritating vapors. Records from a dog with atrial fibrillation exhibit rhythmic fibrillation frequency changes correlated with respiration. In addition, in a morphinized dog with atrial fibrillation, impulses crossed the AV node only during inspiration. The ECGs of a cow and camel exhibit long P-R intervals and biphasic P waves. Finally the elephant ECG shows a clear U wave following the T wave.

Animals↗

Preliminary evaluation of diagnostic tests using horses experimentally infected with trypanosoma evansi.

Seven surra negative horses were intravenously inoculated with 3 x 10(6)Trypanosoma evansi parasites derived from a camel. One horse was maintained as an uninfected negative control. Three antigen and three antibody detection tests were evaluated for diagnosis of infection in horses. The microhaematocrit centrifugation test (MHCT) was the most sensitive, first detecting parasites between one and three days (x 2.4) post infection (p.i.). The antigen (ag)-ELISA detected antigen between three and ten days (x 6.6) p.i. The latex agglutination test (LAT) first gave positive results on day 3 (x 3.0) p.i. Following the treatment of horses with trypanocidal drugs, the MCHT and the mouse inoculation test (MIT) became negative. Antigen levels using LAT declined and reached pre-infection levels in five out of six horses during the period of observation (92-279 days). Antigen levels using the ag-ELISA declined as well but did not reach pre-infection levels in any of the six horses.Three antibody detection techniques, ab-ELISA, card agglutination test (CATT), and immunofluorescent antibody test (IFAT) detected antibodies in the blood of all seven infected horses but not in the uninfected control. However, the ab-ELISA did not discriminate clearly between sera from infected and uninfected horses because unacceptably high ELISA background readings were detected in 15% of the surra negative horses shipped to the UAE from the UK. The ELISA antibody increased above pre-infection levels in the six horses experimentally infected, but not in one horse. In this horse the ELISA antibody level exceeded the cut-off level only after the reoccurrence of the T. evansi infection. The IFAT detected antibodies 15.7 days p.i. in all infected horses.

Animals↗

The electronic states of polyfluorene copolymers with alternating donor-acceptor units.

We calculate the electronic states of the low bandgap polyfluorene-based copolymer DiO-PFDTBT, which consists of alternating 9,9-dioctyl-9H-fluorene and 4,7-di-thiophen-2-ylbenzo[1,2,5]thiadiazole (TBT) units, and compare with the steady-state absorption, emission, and excitation spectrum. Using the semiempirical quantum-chemical (ZINDO) method we can assign the characteristic bands of the "camel-back" absorption spectrum to one charge transfer state at lower energy localized on the TBT unit, and one delocalized excitonic state at higher energy corresponding to the pi-conjugated electron system. Additional "dark" charge transfer states in the gap between these bands have been revealed. Calculations are also made on the red light emitting polyfluorene-based copolymer poly(fluorene-co-benzothiadiazole) (F8BT), which contains benzo[1,2,5]thiadiazole instead of TBT. The nature of the electronic states in F8BT and DiO-PFDTBT are found to be qualitatively the same.

Journal Article↗

Effect of lipid solvents on cutaneous moisture loss.

Application of various lipid solvents and dimethyl sulphoxide to the skin of cattle, sheep, eland and African buffalo exposed to an air temperature of 20 degrees C caused an increase in cutaneous moisture loss. The effect did not occur after exposure to an air temperature of 40 degrees C or after adrenaline administration. Blockage of sweating by either bethanidine or phenoxybenzamine did not affect the response. Camel, donkey, dog and man showed no increase in cutaneous moisture loss after lipid solvent application to the skin. It is concluded that lipid solvents directly stimulate the secretory cells of the sweat glands of members of the family Bovidae and that this is not due to the release of transmitter substance nor to stimulation of adrenergic receptors, but is due to direct stimulation of the sweat glands themselves.

Animals↗

Peaton virus: a new Simbu group arbovirus isolated from cattle and Culicoides brevitarsis in Australia.

A new member of the Simbu group of arboviruses, for which the name Peaton virus is proposed, has been isolated from midges and cattle in Australia. Nine isolates were obtained from 101 pools of the biting midge Culicoides brevitarsis collected at Peachester, Qld, (26.51 degrees S., 152.53 degrees E.) between 30 November and 8 December 1976. Three isolations of the same virus were made from the blood of sentinel cattle collected at Grafton and Tamworth, N.S.W., on 20 January and 13 April 1977, respectively. Peaton virus was shown to be a member of the Simbu group of arboviruses by complement-fixation tests using antisera prepared against Australian strains of Akabane and Aino viruses. It was readily distinguishable from these viruses in cross-neutralization tests in tissue cultures and mice. A serological survey of sentinel cattle showed that neutralizing antibody was detectable only in cattle within the recorded limits of the suspected vector C. brevitarsis. Neutralizing antibody in blood serum was detected in 22 of 157 sheep, 21 of 137 horses, 7 of 18 buffaloes, 7 of 20 goats and 3 of 62 pigs, but not in 22 camels, 34 dogs, 3 cats, 76 human beings, 240 marsupials, 19 reptiles or 31 wild birds. The pathogenecity of Peaton virus has yet to be determined. The Yale Arbovirus Research Unit and the Center for Disease Control, Fort Collins, U.S.A., found that Peaton virus was distinguishable from all other Simbu group viruses and thus is a new virus.

Animals↗

Isolation and characterization of the opioid peptides from rat pituitary: beta-endorphin.

beta-Endorphin was isolated from 200 rat pituitaries by means of high-performance column chromatography, using sensitive fluorometric methods and a radioreceptor assay for opioid activity. The beta-endorphin was characterized as to molecular weight, amino acid composition, and mapping of tryptic peptides by a new microtechnique. It was found that rat beta-endorphin is identical to camel and sheep beta-endorphin. Furthermore, alpha-endorphin, Met-enkephalin, a nonpeptide morphine-like compound (MLC), and an additional unidentified opioid compound were detected in the extract of rat pituitary.

Amino Acids↗

Beta-Endorphin: dissociation of receptor binding activity from analgesic potency.

Biological activities of synthetic camel beta-endorphin and human beta-endorphin (beta h-EP) have been measured by the radioreceptor binding assay, using [Tyr27-3H]-beta h-EP as the primary ligand and by the tail-flick test for analgesic potency. Four synthetic analogs of beta h-EP, namely [Gly31]-beta h-EP-Gly-NH2, [Gly31]-beta h-EP-Gly-Gly-NH2, [Gln8,Gly31]-beta h-EP-Gly-Gly-NH2, and [CH3(CH2)4NH231]-beta h-EP, have also been assayed by the same procedures. Results indicate a clear dissociation of radioreceptor binding activity from analgesic potency.

Amino Acid Sequence↗

Human beta-endorphin: specific binding in neuroblastoma N18TG2 cells.

The mouse neuroblastoma N18TG2 has specific, saturable binding sites for human beta-endorphin (beta h-EP). The affinity and number of sites are 1.1 nM and 280,000 per cell, respectively, beta h-EP binding is not inhibited by [Leu]enkephalin or morphine at concentrations up to 0.1 mM; beta h-EP-(6--31) is a potent inhibitor of binding, and camel beta-EP is much less potent. The data suggest the importance of the nonenkephalin segment of the beta h-EP molecule for interaction with the binding site in N18TG2 cells.

Animals↗

beta-Endorphin: demonstration of a tertiary structure in aqueous solution.

Difference spectra generated during thermolysin digestion of camel beta-endorphin at pH 8.2 or at pH 6.5 indicate rapid blue shifting of the near-UV absorption bands of the NH2-terminal tyrosine. A similar spectral change is not observed for the NH2-terminal tyrosine in [Met]enkephalin when it is digested under similar conditions. These results suggest that enzymatic digestion destroys or alters some structural interaction between the NH2-terminal tyrosyl residue of the endorphin and a residue(s) within the COOH-terminal segment of the molecule. Peptide mapping of the digest as a function of time suggests that cleavage of the bond linking the alanine-21 and isoleucine-22 residues produces most of the observed effect. These data provide evidence for the existence of a tertiary structure for beta-endorphin in aqueous solutions.

Endorphins↗

Corticotropin and beta-endorphin-like materials are native to unicellular organisms.

Multiple molecular forms of immunoreactive corticotropin (ACTH) and beta-endorphin were present in extracts of a unicellular eukaryote (Tetrahymena pyriformis). One form of immunoreactive ACTH reacted similarly with two different ACTH antisera (one specific for the 11-24 sequence and the other with determinants within sequences 1-14 and 17-39) and migrated with synthetic hACTH-(1-39) in a gel filtration system. This form also exhibited ACTH bioactivity in a dispersed rat adrenal cell bioassay system, with a mean immunoassay/bioassay ratio of 1.5. Gel filtration revealed multiple size classes of immunoreactive beta-endorphin; a major peak of radioreceptor activity was detected which exhibited a K(av) similar to that of authentic beta-endorphin. A major portion of immunoreactive beta-endorphin-sized material exhibited retention times similar to those of synthetic human and camel beta-endorphin upon reverse-phase high-pressure liquid chromatography. These distinctive properties and specificities would seem to exclude the presence of limited homologies with sequences present in other proteins. High molecular weight material containing both ACTH and beta-endorphin antigenic determinants was also demonstrated, suggesting, but not proving, the presence of a common precursor molecule.

Journal Article↗

beta-Endorphin: characterization of binding sites specific for the human hormone in human glioblastoma SF126 cells.

The established human glioblastoma cell line SF126 was found to bind tritiated human beta-endorphin (beta h-EP) in a saturable fashion. From displacement studies, the ED50 was estimated to be about 2.5 nM. The Kd was estimated as 1.9 X 10(-9) M and Scatchard analysis showed a biphasic pattern with a predominant low-affinity component. Binding reached a maximum at about 90 min at 22 degrees C and was instantaneously reversible. Tritiated [D-Ala2,D-Leu5]enkephalin and tritiated dihydromorphine did not bind to the cells. Sodium at a concentration of 150 mM decreased the specific binding by 80%. The interaction with the cellular binding site appeared to be mediated by the COOH-terminal segment of beta h-EP, as beta h-EP-(6-31) retained a high potency for displacing tritiated beta h-EP, and beta h-EP-(1-27) has no activity. Camel beta-EP was only about 1% as active as the human hormone.

Binding, Competitive↗

"Peptabody": a new type of high avidity binding protein.

A new type of high avidity binding molecule, termed "peptabody" was created by harnessing the effect of multivalent interaction. A short peptide ligand was fused via a semi-rigid hinge region with the coiled-coil assembly domain of the cartilage oligomeric matrix protein, resulting in a pentameric multivalent binding molecule. In the first peptabody (Pab-S) described here, a peptide (S) specific for the mouse B-cell lymphoma BCL1 surface Ig idiotype, was selected from a phage display library. A fusion gene was constructed encoding peptide S, followed by the 24 aa hinge region from camel IgG and a modified 55 aa cartilage oligomeric matrix protein pentamerization domain. The Pab-S fusion protein was expressed in Escherichia coli in a soluble form at high levels and purified in a single step by metal-affinity chromatography. Pab-S specifically bound the BCL1 surface idiotype with an avidity of about 1 nM, which corresponds to a 2 x 10(5)-fold increase compared with the affinity of the synthetic peptide S itself. Biochemical characterization showed that Pab-S is a stable homopentamer of about 85 kDa, with interchain disulfide bonds. Pab-S can be dissociated under denaturing and reducing conditions and reassociated as a pentamer with full-binding activity. This intrinsic feature provides an easy way to combine Pab molecules with two different peptide specificities, thus producing heteropentamers with bispecific and/or chelating properties.

Animals↗

Phylogenetic relationships among cetartiodactyls based on insertions of short and long interpersed elements: hippopotamuses are the closest extant relatives of whales.

Insertion analysis of short and long interspersed elements is a powerful method for phylogenetic inference. In a previous study of short interspersed element data, it was found that cetaceans, hippopotamuses, and ruminants form a monophyletic group. To further resolve the relationships among these taxa, we now have isolated and characterized 10 additional loci. A phylogenetic analysis of these data was able to resolve relationships among the major cetartiodactyl groups, thereby shedding light on the origin of whales. The results indicated (i) that cetaceans are deeply nested within Artiodactyla, (ii) that cetaceans and hippopotamuses form a monophyletic group, (iii) that pigs and peccaries form a monophyletic group to the exclusion of hippopotamuses, (iv) that chevrotains diverged first among ruminants, and (v) that camels diverged first among cetartiodactyls. These findings lead us to conclude that cetaceans evolved from an immediate artiodactyl, not mesonychian, ancestor.

Animals↗

The gene for a novel member of the whey acidic protein family encodes three four-disulfide core domains and is asynchronously expressed during lactation.

Secretion of whey acidic protein (WAP) in milk throughout lactation has previously been reported for a limited number of species, including the mouse, rat, rabbit, camel, and pig. We report here the isolation of WAP from the milk of a marsupial, the tammar wallaby (Macropus eugenii). Tammar WAP (tWAP) was isolated by reverse-phase HPLC and migrates in SDS-polyacrylamide gel electrophoresis at 29.9 kDa. tWAP is the major whey protein, but in contrast to eutherians, secretion is asynchronous and occurs only from approximately days 130 through 240 of lactation. The full-length cDNA codes for a mature protein of 191 amino acids, which is comprised of three four-disulfide core domains, contrasting with the two four-disulfide core domain arrangement in all other known WAPs. A three-dimensional model for tWAP has been constructed and suggests that the three domains have little interaction and could function independently. Analysis of the amino acid sequence suggests the protein belongs to a family of protease inhibitors; however, the predicted active site of these domains is dissimilar to the confirmed active site for known protease inhibitors. This suggests that any putative protease ligand may be unique to either the mammary gland, milk, or gut of the pouch young. Examination of the endocrine regulation of the tWAP gene showed consistently that the gene is prolactin-responsive but that the endocrine requirements for induction and maintenance of tWAP gene expression are different during lactation.

Amino Acid Sequence↗