Digestive studies with a feed developed for realimentation of starving reindeer

The properties of three different feeds were compared when offered to reindeer calves as single feeds after a starvation period of the 48 h. The feeds were lichen and two pelleted commercial reindeer feeds, RF-71 and RF-80. The two latter differed in concentration of readily digestible carbohydrates (high in RF-71) and in the inclusion of seaweed meal in RF-80. Seven calves were offered the three diets in a latin square design. Measurements involved feed intake and rumen concentrations of volatile fatty acids, ammonia and pH during a five day period after the end of the starvation period. Feeding RF-80 gave rise to higher feed intakes and more rapid normalisation of rumen VFA and ammonia concentration than the other pelleted feed. Rumen pH reached a minimum of 5.4 in animals fed RF-71, while the average minimum pH during the observation period was 6.1-6.2 when RF-80 was given. Inappetance for 1-2 days after refeeding occurred only with RF-71. RF-80 has now replaced RF-71 as the commercial reindeer feed in Norway.Fordøyelsesforsøk med et for utviklet til overgangsforing av sveltende rein.Abstract in Norwegian / Sammendrag: En har sammenlignet egenskapene til tre forskjellige fortyper gitt til reinkalver som eneste for etter en sveltperiode på 48 timer. Fortypene var reinlav og to pelletterte, kommersielle reinfor: RF-71 og RF-80. De siste to adskilte seg fra hverandre i konsentrasjonen av lettfordøyelige karbohydrater (høyest i RF-71) og i innblanding av tangmel i RF-80. Syv reinkalver ble gitt de tre dietter i «latin square» forsøksmønster. Målingene omfattet: forinntak, konsentrasjon i vominnhold av flyktige fettsyrer (VFA) og ammonium samt verdier av pfi gjennom en fem-dagers periode etter avsluttet sveltperiode. Foring med RF-80 økte forinntaket og forårsaket en raskere normalisering av VFA- og ammoniumkonsentrasjonene enn foring med RF-71. pfi nådde et minimum på 5,4 hos dyr som fikk RF-71, mens gjennomsnittlig verdi av pH gjennom observasjonsperioden var 6,1-6,2 når det ble gitt RF-80. Apetittløshet i 1-2 dager etter gjenopptatt foring inntraff bare ved bruk av RF-71. RF-80 har nå erstattet RF-71 som kommersielt for til rein i Norge

Protein purification and gene isolation of chlamysin, a cold-active lysozyme-like enzyme with antibacterial activity

AbstractAn antibacterial ∼11 kDa protein designated chlamysin was isolated from viscera of the marine bivalve Chlamys islandica. Chlamysin inhibited the growth of all Gram-positive and Gram-negative bacteria tested. The isolated protein was highly efficient in hydrolyzing Micrococcus luteus cells only at low pH (4.5–6.2) and at low temperature (4–35°C). No significant loss of enzyme activity was observed after 30 days storage at room temperature or after heating to 70°C for 15 min, suggesting relatively high protein structure stability. Sequence-analyzed fragments of the protein revealed data which guided the isolation of the cDNA gene, encoding a 137 amino acid chlamysin precursor in scallops. The deduced protein contains a high portion of cysteine, serine and histidine residues and has a predicted isoelectric point below 7. The chlamysin protein was found to have sequence homology to an isopeptidase and to a recently published bivalve lysozyme

Germ Line Origin and Somatic Mutations Determine the Target Tissues in Systemic AL-Amyloidosis

BACKGROUND: Amyloid is insoluble aggregated proteins deposited in the extra cellular space. About 25 different proteins are known to form amyloid in vivo and are associated with severe diseases such as Alzheimer's disease, prion diseases and type-2 diabetes. Light chain (AL) -amyloidosis is unique among amyloid diseases in that the fibril protein, a monoclonal immunoglobulin light chain, varies between individuals and that no two AL-proteins with identical primary structures have been described to date. The variability in tissue distribution of amyloid deposits is considerably larger in systemic AL-amyloidosis than in any other form of amyloidosis. The reason for this variation is believed to be based on the differences in properties of the amyloidogenic immunoglobulin light chain. However, there is presently no known relationship between the structure of an AL-protein and tissue distribution. METHODOLOGY/PRINCIPAL FINDINGS: We compared the pattern of amyloid deposition in four individuals with amyloid protein derived from variable light chain gene O18-O8, the source of a high proportion of amyloidogenic light chains, and in whom all or most of the fibril protein had been determined by amino acid sequencing. In spite of great similarities between the structures of the proteins, there was a pronounced variability in deposition pattern. We also compared the tissue distribution in these four individuals with that of four other patients with AL-amyloid derived from the L2-L16 gene. Although the interindividual variations were pronounced, liver and kidney involvement was much more evident in the latter four. CONCLUSIONS/SIGNIFICANCE: We conclude that although the use of a specific gene influences the tissue distribution of amyloid, each light chain exhibits one or more determinants of organ-specificity, which originate from somatic mutations and post-translational modifications. Eventual identification of such determinants could lead to improved treatment of patients with AL amyloidosis

Digestive studies with a feed developed for realimentation of starving reindeer

The properties of three different feeds were compared when offered to reindeer calves as single feeds after a starvation period of the 48 h. The feeds were lichen and two pelleted commercial reindeer feeds, RF-71 and RF-80. The two latter differed in concentration of readily digestible carbohydrates (high in RF-71) and in the inclusion of seaweed meal in RF-80. Seven calves were offered the three diets in a latin square design. Measurements involved feed intake and rumen concentrations of volatile fatty acids, ammonia and pH during a five day period after the end of the starvation period. Feeding RF-80 gave rise to higher feed intakes and more rapid normalisation of rumen VFA and ammonia concentration than the other pelleted feed. Rumen pH reached a minimum of 5.4 in animals fed RF-71, while the average minimum pH during the observation period was 6.1-6.2 when RF-80 was given. Inappetance for 1-2 days after refeeding occurred only with RF-71. RF-80 has now replaced RF-71 as the commercial reindeer feed in Norway. Fordøyelsesforsøk med et for utviklet til overgangsforing av sveltende rein. Abstract in Norwegian / Sammendrag: En har sammenlignet egenskapene til tre forskjellige fortyper gitt til reinkalver som eneste for etter en sveltperiode på 48 timer. Fortypene var reinlav og to pelletterte, kommersielle reinfor: RF-71 og RF-80. De siste to adskilte seg fra hverandre i konsentrasjonen av lettfordøyelige karbohydrater (høyest i RF-71) og i innblanding av tangmel i RF-80. Syv reinkalver ble gitt de tre dietter i «latin square» forsøksmønster. Målingene omfattet: forinntak, konsentrasjon i vominnhold av flyktige fettsyrer (VFA) og ammonium samt verdier av pfi gjennom en fem-dagers periode etter avsluttet sveltperiode. Foring med RF-80 økte forinntaket og forårsaket en raskere normalisering av VFA- og ammoniumkonsentrasjonene enn foring med RF-71. pfi nådde et minimum på 5,4 hos dyr som fikk RF-71, mens gjennomsnittlig verdi av pH gjennom observasjonsperioden var 6,1-6,2 når det ble gitt RF-80. Apetittløshet i 1-2 dager etter gjenopptatt foring inntraff bare ved bruk av RF-71. RF-80 har nå erstattet RF-71 som kommersielt for til rein i Norge

Articular, monoclonal gamma3 heavy-chain deposition disease: characterization of a partially deleted heavy-chain gene and its protein product synthesized in vivo and in vitro.

Objective A patient presented with heavy-chain deposition disease (HCDD), exhibiting severe erosive polyarthropathy caused by synovial deposits of abnormal monoclonal, heavily deleted free 3 heavy chains lacking the VH and CH1 domains. The absence of VH was surprising, since it is considered important for pathogenic tissue deposition. This study was undertaken to analyze the genetic structure of the heavy chain, the protein product synthesized in vitro, and that deposited in the synovium in comparison with the serum and urinary proteins. Methods Hybridomas were made by fusion of blood and bone marrow mononuclear cells with mouse myeloma cells. Cloned B cell hybridomas secreting 3 were selected and analyzed by polymerase chain reaction. Purified hybridoma Ig was sequenced by Edman degradation. Antiserum raised to a peptide corresponding to residues 2-15 of the truncated VH was used in Western blots of synovial tissue. Results The hybridomas secreted free 3 chains consisting of a VH4 gene truncated 21 nucleotides into the first complementarity-determining region and then reading straight into the hinge region. The amino acid sequence confirmed the presence of residues 1-32 of the VH4 gene. Immunoblotting of synovial tissue showed the presence of Ig with truncated VH. Conclusion The 3 heavy chain had a deletion of VH from codon 33 and of the entire CH1. In vivo, the 32 VH amino acids were proteolytically degraded. In the joint, however, the 32 residues of VH remained intact, consistent with a pathogenic role of VH for tissue deposition. To our knowledge, this is the first reported case of HCDD causing an erosive, polyarticular arthropathy as the dominating clinical feature

Elucidation of the primary and three-dimensional structure of the uterotonic polypeptide kalata B1

The amino acid sequence and structure of a uterotonic polypeptide extracted from the African plant Oldenlandia affinis DC have been determined. The peptide, kalata B1, consists of 29 amino acid residues and is rich in cysteine (6), threonine (5), and glycine (5), Enzyme cleavage studies show that the polypeptide backbone is cyclic. The three-dimensional solution structure has been determined using two-dimensional nuclear magnetic resonance (NMR) spectroscopy and distance-restrained simulated annealing, Kalata B1 is composed mainly of beta-strands connected by tight turns, forming regions of beta-sheet, except in the case of one section which forms a longer, less structured loop. The tertiary fold, together with the disulfides that form a sulfur core, produces a striking and unusual surface in which the majority of the hydrophobic residues form. a solvent-exposed patch. The hydrophobic side of kalata B1 is flanked by two diametrically opposed and opposite-charged residues. The structure calculations have been used to predict the previously unknown disulfide bond connectivities using two approaches. In the first, a family of structures was calculated on the basis of NOE constraints without the assumption of a specific disulfide connectivity. The resultant structures were examined to determine whether the calculated position of the sulfur atoms suggested that one set of disulfide connectivities was more likely than the other, theoretically possible, sets. In the second approach, a separate family of structures (50 per set) was calculated for each of the 15 possible disulfide-bonded molecules. The resultant families of structures were compared to see whether one was favored over the others, Both approaches led to the same global fold, and the most likely disulfide connectivity is predicted to be 5-22, 13-27, and 17-29. In the calculated structure the cyclic peptide backbone is folded back onto itself and braced with disulfide pairs across diagonally opposed beta-strands, This structure involves one of the disulfide bonds (5-22) threading through the eight amino acid loop formed by the other two disulfide bonds and the peptide fragments connecting them

Cleavage of AL amyloid proteins and AL amyloid deposits by cathepsins B, K, and L

Cathepsin (Cath) B, CathK and CathL are cysteine proteases that participate in the lysosomal protein degradation system and are expressed in macrophages, epithelioid cells, and multinucleated histiocytic giant cells (MGCs). Both macrophages and MGCs are commonly found adjacent to immunoglobulin light chain -associated (AL) amyloid deposits, which raised the question of whether cysteine proteases are able to cleave AL amyloid proteins and AL amyloid deposits. The present study has investigated whether recombinant human CathB, CathK, and CathL are able to degrade AL(VlambdaVI) amyloid proteins and AL amyloid deposits. Using immunohistochemistry, CathB, CathK, and CathL were found adjacent to AL amyloid deposits. In vitro degradation experiments using purified AL amyloid proteins showed that CathB, CathK, and CathL degrade AL(VlambdaVI) amyloid proteins. Furthermore, using unfixed tissue sections from an amyloidotic spleen as an in vitro model for extracellular proteolysis of intact amyloid deposits, it was demonstrated that all three cysteine proteases are also capable of degrading AL amyloid in situ. This is the first study to show that cysteine proteases are able to cleave AL amyloid proteins. However, the efficiency with which proteolysis occurs depends on the concentration of active protease recruited at the sites of amyloid deposition, and possibly on the structure of the AL amyloid proteins. Copyright 2004 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, LtdEnglish15095475NRC publication: Ye