Primary structure of Beijing duck apolipoprotein A-1

The primary structure of Beijing duck apolipoprotein A-1 was determined by sequencing peptide fragments derived from tryptic and endoproteinase Asp-N digestion of the protein, and alignment with homologous chicken apo A-1. All of the peptide fragments were isolated by high-pressure liquid chromatography (HPLC) with a Vydac C18 column using a trifluoroacetic acid (TFA) buffer system. The N-terminus of the protein was determined to be aspartic acid by directly sequencing 52 residues of the intact protein. The C-terminus was alanine. The protein contains 240 amino acid residues. By analysis of the whole protein and its tryptic peptides, a six amino acid (Arg-Tyr-Phe-Trp-Gln-His) prosegment was determined. No cross-reactivity between duck and human apo A-1 with a goat antiserum against human apo A-1 was found. Sequence analysis of apo A-1 of other species indicates that amino acid substitutions in rat are more extensive than in other mammals. Isoleucine residues in apo A-1 are inversely correlated to the homology of human to other species, except dog.     

An isometric predictor for maximum acceptable weight of lift for Chinese men

The aim of this study was to examine the practicality of the modified isometric strength tests to predict the maximum acceptable weight of lift (MAWL) of Chinese men. The modified strength tests allow the participant to pull on the load cell in front of the body and to apply force in a functional free posture. Both the modified and the standard strength data of each participant were used as predictors for the MAWLs. The prediction models were constructed and evaluated under task conditions of two lifting ranges, two box sizes, and three lifting frequencies. To realize the effect of modifications, testing posture was recorded and the joint angles were calculated. A stepwise multiple regression analysis indicated that modified composite strength (MCS), chest circumference, and acromial height accounted for 86% to 91% of the variance. Because the strength of the upper extremity body was also recruited in the test, the weak upper extremity strength of the Chinese participants would therefore be better reflected. Evidence for the existence of a close match between MAWL and MCS values, as well as the task conditions for its existence, suggest that a simple isometric strength measure is a good predictor for the MAWL.     

Sesquiterpenes from the leaves of Tithonia diversifolia

Two new compounds, a germacrane sesquiterpene, 1-acetyltagitinin A (1), and a guaianane sesquiterpene, 8beta-isobutyryloxycumambranolide (2), were isolated from leaves of Tithonia diversifolia, together with two known compounds, methyl 3alpha-acetoxy-4alpha-hydroxy-11(13)-eudesmen-12-oa te and tagitinin A. The structures of compounds 1 and 2 were elucidated on the basis of spectral and chemical evidence.     

Enhancement of human platelet aggregation and secretion induced by rapamycin

BACKGROUND: Rapamycin is a new immunosuppressive drug of the macrolide type. Despite binding to one of the FK-binding proteins as the initial step in intracellular action, further effects differ from those of the other fungally derived macrolides, cyclosporine and tacrolimus. We have previously demonstrated an enhancement of agonist-mediated platelet activation by cyclosporine and tacrolimus which was associated with increased phosphorylation of two intracellular platelet proteins, p20 and p40. Because rapamycin utilizes the same class of binding proteins as tacrolimus, but its action is not associated with the inhibition of calcineurin, we postulated that if the stimulatory effect of cyclosporine or tacrolimus was due to calcineurin inhibition, rapamycin should not affect platelets in a similar fashion. METHODS: Normal, washed human platelets were treated with various concentrations of rapamycin (from ng to microg/ml), and pre-incubated at 37 degrees C with rapamycin for various periods (1-30 min). Several platelet functional parameters were measured in samples treated with rapamycin and these parameters were compared with control platelet samples treated with the vehicle for the same period. Platelet aggregations following exposure to ADP or to the thrombin equivalent, TRAP-6, were measured as changes in optical transmission in a Chronolog lumi-aggregometer. Each experiment was repeated at three or more times and the mean results were used for statistical comparison. RESULTS: Rapamycin-treated platelets demonstrated an increase in their dose- and time-dependent sensitivity to ADP, resulting in a significantly enhanced primary wave of ADP-induced platelet aggregation followed by a secondary wave of aggregation, indicative of granule secretion. Furthermore, rapamycin-treated platelets showed significantly enhanced sensitivity to TRAP-6 as demonstrated by an increase in the initial velocity of aggregation, an increase in their maximal extent of aggregation and an enhancement of granular ATP secretion. Concentrations of rapamycin in the ng range, as well as short pre-incubation times (within min), were sufficient to cause significant enhancement of agonist-induced platelet aggregation and secretion (P < 0.001) as compared with their vehicle controls. CONCLUSIONS: Rapamycin significantly potentiates agonist-induced platelet aggregation in a time- and dose-dependent manner. As these findings are similar to those observed with the other fungal macrolides, we hypothesize that inhibition of calcineurin may not be necessary for the increase in intracellular protein phosphorylation observed following exposure of platelets to cyclosporine or tacrolimus. Whether the rapamycin-induced enhancement of sensitivity to agonists and platelet hyperaggregability explains the thrombocytopenia observed in patients when high doses of rapamycin are administered in the clinical setting, and whether these effects are synergistic with cyclosporine, are questions which remain to be investigated.     

Photoageing-associated mitochondrial DNA length mutations in human skin

It has recently been suggested that mitochondrial DNA (mtDNA) mutations are important contributors to human ageing and degenerative diseases. Using PCR techniques, we demonstrated three types of mtDNA length mutations, a 4977 bp deletion, a 7436 bp deletion and tandem duplications, in normal human skin tissues. We found that these mutations started to appear in the third decade of life, and the age at which the mutations could be detected in sun-exposed skin was usually younger than in non-exposed skin. Moreover, the incidences of these deletions and tandem duplications of mtDNA in sun-exposed skin were all significantly higher than those in non-exposed skin (P < 0.05). The 4977 bp deletion was the most prevalant mtDNA mutation in human skin, and the 7436 bp deletion was the least frequent among the three types of mtDNA mutations examined. We first demonstrated the existence of tandem duplications with sizes of about 260 bp, 200 bp and 150 bp in the D-loop region of mtDNA in the skin of elderly individuals. Among the three tandem duplications, the 200-bp duplication was found to occur most frequently in ageing skin. The tandem duplications were found to coexist with either or both of the deletions in some elderly individuals. The frequency of occurrence of mtDNA deletions and tandem duplications in skin was found to increase in an age-dependent manner. However, the incidence of tandem duplications was not well correlated with the age of the subject.(ABSTRACT TRUNCATED AT 250 WORDS)     

Advanced renal cell carcinoma: treatment with xenogeneic immune ribonucleic acid and appropriate surgical resection

Herein we describe the first clinical treatment of renal cell carcinoma in humans with xenogeneic immune ribonucleic acid. Twelve patients with advanced renal cell carcinoma have been treated by appropriate operations to remove tumor bulk followed by specific passive immunotherapy. Xenogeneic specific immune ribonucleic acid was prepared from the spleen of normal sheep that had received 4 weekly injections of a homogenate of renal cell carcinoma. Results indicate that 1) xenogeneic specific immune ribonucleic acid can safely be given to humans without local or systemic toxicity, 2) there is a suggestion of clinical benefit, since only 2 patients have had progression of known metastases during treatment with immune ribonucleic acid and 3) xenogeneic immune ribonucleic acid can enhance the immune response to renal cell carcinoma, as demonstrated by in vitro lymphocytoxicity tests.