Andamino Acid Research Articles

Cloning and sequencing of the canine and feline cardiac troponin I genes

To determine the gene sequences of canine and feline cardiac troponin I (cTnI), express the protein from the cloned gene in vitro, and validate the use of a commercial cTnI serum analyzer in these species via detection of the expressed protein or comparison of sequence homology. Samples of ventricular myocardium from 5 healthy adult mixed-breed dogs and 5 healthy adult domestic shorthair cats. The RNA was extracted from myocardial samples, and cDNA was synthesized via reverse transcriptase polymerase chain reaction and sequenced. The canine cDNA for the coding region was expressed in cell culture and analyzed by western blot and sandwich enzyme-linked immunosorbent assays. Canine and feline cTnI genes were cloned and sequenced. Homology of the nucleotide and amino acid sequences of the canine and feline cTnI genes with human and rodent cTnI genes were high; the greatest homology was detected between canine and feline genes (95% and 96%, respectively). Recombinant canine cTnI protein was detected by a commercial serum cTnI analyzer and by western blot analysis. Results indicated that commercial cTnI analyzers can be used to measure serum cTnI concentration from dogs and cats. Additionally, our preliminary characterization of the feline cTnI gene may facilitate further investigation of cTnI and its role in familial hypertrophic cardiomyopathy in cats.

Identification of a phosphopeptide in bovine alpha s1-casein digest as a factor influencing proliferation and immunoglobulin production in lymphocyte cultures.

Digestion of bovine alpha s1-casein with bovine trypsin produced peptide(s) with an inhibitory effect on concanavalin A-induced proliferation of mouse spleen cells. One of these peptides was isolated from the alpha s1-casein digest following ultrafiltration, hydroxyapatite chromatography and reversed-phase HPLC, and amino acid composition and sequence analyses showed it to be sequence 59-79 from the phosphoserine-rich region of alpha s1-casein. The isolated peptide significantly inhibited the concanavalin A-induced proliferation of mouse spleen cells and rabbit Peyer's patch cells, whereas it enhanced the lipopolysaccharide- and phytohaemagglutinin-induced proliferation of both cells. The peptide displayed mitogenic activity in the cell cultures without the commercial mitogen, and significantly enhanced immunoglobulin production. Moreover, residues 1-25 from the phosphoserine-rich region of bovine beta-casein had a similar effect on the proliferation of mouse spleen cells and rabbit Peyer's patch cells stimulated or not stimulated by the commercial mitogen. These results indicate that caseinophosphopeptides may act as a humoral immunostimulator in cell cultures.

Muscle and plasma amino acids and nutritional status in kidney-transplanted children

Nutritional status, assessed by anthropometric and biochemical methods, protein and amino-acid (AA) composition and muscle water were evaluated in 11 kidney transplanted children and in a control group of 10 children with normal renal function who were undergoing elective surgery. Samples of the rectus abdominis muscle were taken when surgery was performed in the control children and when the peritoneal catheter was removed in the transplanted children. The mean time from the transplantation to the study time was 97 +/- 14 days (range 72-114 days). Height was reduced in the transplanted children compared to the controls but skinfold thickness, arm muscle circumference and serum proteins (total protein, albumin, transferrin, pseudocholinesterase) were normal. The body mass index was over the 50 degree percentile in nine of the eleven children. The muscle contents of total, extracellular, and intracellular water, alkali-soluble protein (ASP), DNA and the ASP/DNA ratio were not significantly different in transplanted children from those in the controls. Plasma leucine and taurine levels were significantly decreased, whereas plasma citrulline and alanine levels and the glycine/serine ratio were increased. Muscle threonine, alanine+taurine, glycine and aspartic acid levels as well as the glycine/serine ratio were increased in the transplanted children. Transplanted children show an almost normal muscle AA profile and a plasma AA pattern that seems to be related to the prednisone therapy.

Effects of Keto-analogues on Phosphocalcic and Aminoacid Metabolism in Dialysed Patients: A Crossover Study

It has been suggested that calcium salts of keto-analogues (KA) have beneficial effects on phosphocalcic and aminoacid (AA) metabolism. To confirm this, their short-term effects were evaluated on chronic dialyzed patients. In a prospective, randomised, crossover study, eight and seven patients were put on KA (200 mg/kg/d) and assigned either a low-protein diet (LP:0.4 g/kg/d) or a normal one (NP: 1 g/kg/d) for 15 days. The two treatments were interchanged after 15 days of washout. KA.LP was accompanied by: a) decreases in calorie intake (12%; p = 0.001) and in blood concentrations of albumin (5%, p = 0.004), urea (32%, p = 0.001), phosphate (29%, p = 0.001), parathormone (27%, p = 0.008), isoleucine (24%, p = 0.04), 1 and 3 methyl-histidine (71%, p = 0.03; 24%, p = 0.005), valine (19%, p = 0.004) and hydroxyproline (85%, p = 0.009); b) increases in calcemia (9%, p = 0.002), cystathionine (991%, p = 0.0001) and threonine (22%, p = 0.04). KA.NP was accompanied by: a) decreases in phosphatemia (15%, p = 0.03) and parathormone (18%, p = 0.06); b) increases in calcemia (9%, p = 0.002), cystathionine (427%, p = 0.0001), and phenylalanine (28%, p = 0.013). Calcium salts of keto-analogues together with a low or normal protein diet thus seem to reduce blood concentrations of phosphates and parathormone, and raise calcium; however their action on aminoacids needs further investigation.

Photoreductive decarboxylation of carboxylic acids via their benzophenone oxime esters

Decarboxylation of various carboxylic acids, including ∝β- andamino acids, is performed by the photolysis of their benzophenoneoxime esters in isopropanol in the presence of t-butyl mercaptane,and corresponding decarboxylated products are obtained in goodyields under mild photolytic conditions.

14C fixation, metabolic labeling patterns, and translocation profiles during leaf development in Populus deltoides

The incorporation of photosynthetically fixed (14)CO2 and the distribution of (14)C among the main chemical constituents of laminae and petioles were examined in cottonwood (Populus deltoides Bartr. ex Marsh.) leaves ranging in age from Leaf Plastochron Index (LPI) 3 (about one-quarter to one-third expanded) to LPI 30 (beginning of senescence). In addition, carbon flow among chemical fractions and translocation from leaves of LPI 7 and 14 were examined periodically up to 24 h after labeling. Specific activity of (14)C (on dry-weight basis) increased in developing laminae to full leaf expansion, decreased in the mature leaves to LPI 16, then remained constant to LPI 30. In developing leaves (LPI 3-5), after 2 h, most of the (14)C was found in protein, pigments, lipids, and other structural and metabolic components necessary for cell development; only 28% was in the sugar fraction of the lamina. In fully expanded leaves (LPI 6-8), after 2 h, the sugar fraction contained 50-60% and about 90% of fixed (14)C in the lamina and the petiole, respectively. In a pulsechase "kinetic series" with recently mature leaves, 60% of the (14)C was found in the sugar fraction after 15 min of (14)CO2 fixation. Over the 24-h translocation period, (14)C decreased in sugars to 23% and increased in the combined residue fraction (protein, starch, and structural carbohydrates) to about 60% of the total activity left in the lamina. Within 24 h after labeling, the turnover of (14)C-organic acids,-sugar, and-amino acids (either metabolzed or translocated from the leaf) was 30, 70 and 80%, respectively, of that initially incorporated into these fractions by a leaf at LPI 7 (turnover was 55% of (14)C-organic acids, 80% of (14)C-sugar, and 95% of (14)C-amino acids at LPI 14). Anatomical maturity in cottonwood leaves is closely correlated with physiological maturity and with production of translocatable sugar.

EFFECTS OF HETEROTROPHIC AND AUTOTROPHIC GROWTH CONDITIONS ON THE COMPOSITION OF Chlorella sorokiniana

SUMMARY –The effects of heterotrophic and autotrophic growth conditions on the composition of Chlorella are discussed in relation to the potential food value of algae. Cells from the two culture modes differ appreciably in proximate analysis and elemental composition; however, fiber content and vitamin andamino acid composition are essentially unchanged. On this basis it appears that heterotrophic algae, which are less expensive to produce, can be used to examine the acute toxicity problem with bacteria‐free cells. In addition, heterotrophic algae may also be used to predict results that would be obtained with autotrophic material in studies of processing techniques designed to improve protein quality and digestibility.

Nutritional disorders of ruminants.

Metabolic fuels of the ruminant consist mainly of volatile fatty acids produced in the rumen by microbial digestion of dietary polysaccharides and amino acids derived from dietary protein. As almost no glucose is obtained directly from its food, any specific requirements for this sugar are met by synthesizing it from non-carbohydrate sources: principally propionate and the glucogenic amino acids.