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Biomedical subjects

S Miyatani

Publications and source records attributed to S Miyatani.

36 records · Page 2Linked to original sources

Synthesis and degradation of collagens in skin of healthy and protein-malnourished rats in vivo, studied by 18O2 labelling.

To explore the effects of growth retardation, caused by restricted protein intake, on collagen turnover in the whole skin, Sprague-Dawley rats (n = 20) were labelled with 18O2 and fed on either an adequate (18%) or a low (3%) lactalbumin diet. Skin biopsies were obtained at intervals during the following 6 months. Independent groups of animals (n = 186) were used to determine the size of the 0.5 M-acetic acid-soluble and -insoluble collagen pools in the entire skin of healthy and malnourished rats. Collagen was estimated by measurement of hydroxyproline. Soluble-collagen synthesis rates were equivalent to 99 +/- 8 mumol of hydroxyproline/day in healthy animals and 11 +/- 2 mumol/day in malnourished rats. Insoluble-collagen synthesis rates were 32 and 5 mumol of hydroxyproline/day in the healthy and protein-depleted rats respectively. The degradation of soluble collagen amounted to 37 +/- 8 and 6 +/- 2 mumol of hydroxyproline/day in the healthy and malnourished groups respectively. Efflux of collagen from the soluble collagen, defined as the sum of the rate of soluble collagen that is degraded plus that which matures into insoluble collagen, was 70 +/- 8 and 11 +/- 2 mumol of hydroxyproline/day in the healthy and malnourished groups respectively. Insoluble collagen was not degraded in either group. The fraction of soluble collagen leaving the pool that was converted into insoluble collagen was 0.46 in both diet groups. It is concluded that the turnover of soluble collagen is markedly decreased with malnutrition, but degradation and conversion into insoluble collagen account for the same proportions of efflux from the soluble-collagen pool as in rapidly growing rats.

Animals↗

Responses of sodium balance, blood pressure, and other variables to sodium loading in Papua New Guinea highlanders.

For determination of the responses of sodium balance, blood pressure, and other relevant variables to Na loading in people with a low intake of Na, 10 male Papua New Guinea highland subjects were given additional Na at two levels (128 and 256 mmol/d) for 10 d after a 3-d control period of low-Na diet. Na loading caused a marked positive balance of Na, decreases of aldosterone concentration and renin activity in the plasma, and a decrease of urinary aldosterone excretion. The blood pressure, particularly that measured at noon, increased in the latter half of the Na-loading period, the increase being significant in the group given 256 mmol of sodium daily: the systolic and diastolic blood pressure increased from 92 +/- 8 over 56 +/- 7 mm Hg in the control period to 102 +/- 7 over 60 +/- 4 mm Hg in the latter half of the test period (p less than 0.05).

Adult↗

Effects of low protein intake on protein metabolism of Papua New Guinea highlanders studied with [15N]glycine.

The effects of low protein intake on protein metabolism, including the size of pools and the protein synthesis rates, were studied by use of [15N]glycine in Papua New Guinea highlanders. Studies were made on 9 men between October and December in 1982. In experiment 1, two subjects were given a protein-free diet (PFD) containing 49.1 kcal/kg of energy. In experiment 2, subjects were given a sweet-potato diet (SPD) containing 45.4 kcal/kg of energy and 0.507 g/kg of protein for 8 days, and then were given a low-protein sweet-potato diet (LPSPD) containing 50.0 kcal/kg of energy and 0.265 g/kg of protein. During the SPD period, the sizes of the metabolic and active protein pools (mean +/- SD) were 270 +/- 134 mgN/kg and 362 +/- 107 mgN/kg, respectively, and the rates of active and inactive protein synthesis were 463 +/- 161 mgN/kg/day and 299 +/- 38 mgN/kg/day, respectively. During the LPSPD period, the sizes of the metabolic pool and active protein pool were 131 +/- 64 mgN/kg and 378 +/- 106 mgN/kg, respectively, and the rates of active and inactive protein synthesis were 490 +/- 206 mgN/kg/day and 280 +/- 26 mgN/kg/day, respectively. The protein metabolism in the LPSPD showed no significant difference from the SPD. The results suggest that, when the energy levels were approximately the same, protein metabolism in Papua New Guinea highlanders was maintained in spite of the decrease in protein intake.

Adult↗

Stage-specific keratins in Xenopus laevis embryos and tadpoles: the XK81 gene family.

This report describes the isolation and characterization of genomic and cDNA clones which define a subfamily of type I keratins in Xenopus laevis whose expression is restricted to embryonic and larval stages. The XK81 subfamily, named after the prototype cDNA clone DG81, contains four members arranged in two pairs of closely homologous loci; they were named 81A1, A2, B1, and B2. Genomic clones were obtained representing all of these regions. The A1 gene has been completely sequenced together with approximately 1 kb of flanking sequences at each end; this gene corresponds to the previously reported cDNA clone 8128 (Jonas, E., T. D. Sargent, and I. B. Dawid, 1985, Proc. Natl. Acad. Sci. USA, 82:5413-5417). The B2 gene is represented by a partial cDNA clone, DG118. Upstream sequences and about half of the coding regions have been sequenced for the B1 and B2 genes, whereas the A2 locus has been identified on the basis of hybridization data and could be a gene or pseudogene. Genomic Southern blotting indicates that all members of the subfamily have been isolated. The keratin proteins encoded by the B1 and B2 genes are 96% homologous in the central rod domain, whereas A/B gene homology in this region is 81%. During development mRNAs derived from A and B genes accumulate coordinately during gastrula and neurula stages; in the tadpole, 81A mRNA decays rapidly, whereas 81B mRNA shows a second abundance peak, persists for most of tadpole life, and decays by metamorphosis. RNAs derived from the XK81 keratin subfamily are undetectable in the adult, where different type I keratin genes are expressed.

Animals↗

Studies of nitrogen balance in male highlanders in Papua New Guinea.

Nitrogen metabolism in Papua New Guinea highlanders was studied by examining their nitrogen balance when they ate their usual diet (experiment 1) or an experimental diet (experiment 2). Studies were made on 39 male highlanders between October and December in 1980 and 1982. In experiment 1, the average protein and energy intakes (mean +/- SD) for three consecutive days of usual diets were 116.2 +/- 31.2 mg N/kg body weight and 46.9 +/- 8.9 kcal/kg body weight in 1980 and 143.5 +/- 54.8 mg N/kg body weight and 47.3 +/- 10.5 kcal/kg body weight in 1982, and thus the nitrogen balances were 27.7 +/- 28.3 and 18.4 +/- 24.2 mg N/kg body weight, respectively. Most dietary protein and energy was derived from sweet potatoes and other vegetable foods, but between 1980 and 1982 the diet changed slightly with increased consumption of imported foods such as rice and canned fish. In the second experiment, when 10 highlanders were given adequate protein diet (about 200 mg N/kg body weight and about 44 kcal total energy/kg body weight for 13 d, much of the dietary nitrogen was retained in their bodies (45.1 +/- 19.3 mg N/kg body weight). When another group of men were fed a sweet-potato diet (81.1 mg N/kg body weight for 8 d followed by a low protein sweet-potato diet containing only 42 mg N/kg body weight with 50 kcal/kg body weight for 14 d, nitrogen true digestibility and net protein utilization increased and their nitrogen balance showed no significant change (-11.0 +/- 5.6 mg N/kg body weight).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Closed-circuit metabolic system with multiple applications.

A closed-circuit metabolic system has been designed and tested for multiple applications. Air pressure within a closed chamber is regulated electronically while allowing for respiratory gas exchange. Compared with a previously reported standard indirect calorimetry system, the new device had by virtue of longer duration of measurement improved precision (coefficient of variation 3% vs. 14%) during studies of O2 consumption both at room temperature and at 5 degrees C. In addition, a more physiological atmospheric environment is maintained. This system has also been utilized for simultaneously labeling groups of up to 20 weanling rats with 18O2 over a 2-day period and for exposure of rats to a hyperoxic (84% O2), normobaric environment for 4-day periods. Potential applications include maintenance of pressure (hypobaric through hyperbaric) and O2 (hypoxic through hyperoxic) controlled environments, exposure to toxic gases, study of diurnal variations in metabolic rate, measurement of metabolic expenditure with activity, and adaptation to other species including humans.

Animals↗

Effect of dietary protein levels on urea utilization in Papua New Guinea highlanders.

Ability to utilize urea nitrogen for body protein synthesis was examined with Papua New Guinea (PNG) highlanders and Japanese (JPN). Eight male PNG highlanders and 8 male JPN were fed on a low protein diet containing 0.55 g protein/kg or an adequate protein diet containing 1.34 g protein/kg for 1 or 2 weeks. The fate of 15N was measured after oral administration of 15N-labelled urea. There was no difference in 15N incorporation into serum protein between PNG highlanders and JPN receiving low protein diets. On the other hand, on the adequate protein diet, 15N incorporation in PNG highlanders was similar to that on the low protein diet, in contrast to that in JPN which was hardly detected in the adequate protein diet. When PNG highlanders take more protein than protein in their usual diet, they effectively incorporate ingested protein into their body protein and urea nitrogen is utilized for synthesis of body protein.

Adolescent↗

Blood status and serum free amino acids in Papua New Guinea highlanders.

Hematological values and serum amino-acid concentrations were measured in 17 healthy male adult Papua New Guinea highlanders who live on a sweet-potato staple diet. Hematological values were within the normal range, except for a low serum urea concentration. The concentrations of serum threonine, valine, isoleucine, leucine and tyrosine were significantly lower, and those of arginine, glutamate, glycine and alanine were significantly higher, than in Japanese controls. These amino acid patterns in the serum of Papua New Guinea highlanders are an indication of low protein nutrition and adequate energy supply. Some essential amino acid and urea concentrations in the serum of nine Papua New Guinea subjects fed on an adequate protein diet (1.3 g/kg body weight, about twice their habitual diet) for 13 days were significantly increased but were still significantly lower than those of Japanese subjects. Serum alanine decreased on an adequate protein diet. These results show that amino acid uptake and utilization by peripheral tissues may be accelerated on an adequate protein diet. Blood status and serum amino acid concentrations did not show any change except for urea and some amino acids, when five Papua New Guinea highlanders were fed on a low protein diet (0.6 g/kg) consecutively for 13 days.

Adolescent↗

Gene expression in Xenopus embryogenesis.

This article considers some aspects of the storage of macromolecules in the oocyte of Xenopus laevis and the activation of previously unexpressed genes during early embryogenesis. The large quantity and complex nature of poly(A)+ RNA accumulated in the egg provides the cleavage embryo with a supply of mRNA sufficient to sustain protein synthesis for several hours of development. Onset of gene activity at the midblastula transition (MBT) leads to the synthesis and accumulation of molecules of various RNA classes, including tRNAs, rRNAs, mRNAs and mitochondrial RNAs. At gastrulation the poly(A)+ RNA population is still qualitatively similar to that of the egg but some sequences not present in egg RNA have accumulated by this time. Through the use of a subtractive cDNA cloning procedure we have prepared a library of sequences that represent genes activated for the first time between MBT and gastrula. A study of several of these cDNA clones suggests that genes in this class are restricted in their activity to embryonic and tadpole stages.

Animals↗

Endogenous nitrogen excretion in male highlanders of Papua New Guinea.

Endogenous nitrogen excretion was examined in highlanders of Papua New Guinea. Eight highlanders were fed a semisynthetic protein-free diet with about 49 kcal/kg of energy for 11 days. At the end of this period they excreted 29.2 +/- 4.0 mg N/kg of urinary nitrogen and 13.9 +/- 2.5 mg N/kg of fecal nitrogen per day. When the values were expressed per basal metabolic rate (BMR), they were 1.25 +/- 0.19 mg N/kcal BMR for urine and 0.59 +/- 0.08 mg N/kcal BMR for feces. The total amount was calculated as 43.1 +/- 4.3 mg N/kg (1.84 +/- 0.18 mg N/kcal BMR). Endogenous urinary nitrogen excretion in highlanders was the lowest so far reported; the effects of ethnic and nutritional backgrounds on obligatory urinary N loss were examined but no clear explanation was found for the highlanders' low value.

Adult↗

Structural analysis of antigen-specific Ia-bearing regulatory T-cell factors: gel electrophoretic analysis of the antigen-specific augmenting T -cell factor.

An antigen-specific T-cell factor (TaF) that specifically augments the antibody response was purified and biochemically analyzed by NaDodSO4/polyacrylamide gel electrophoresis and isoelectric focusing. Biosynthetically labeled TaF was separated from the Nonidet P-40 extract of T-cell hybridoma FL10, which produces a keyhole limpet hemocyanin-specific TaF, by affinity chromatography either with antigen or with monoclonal anti-I-A antibodies. The material thus obtained was composed of two different types of molecules of molecular weights of 67,000 and 33,000 under nonreducing conditions. After reduction with dithiothreitol, all the molecules migrated to the position of molecular weight 33,000. The absorption studies with immunoadsorbents of antigen and antibodies revealed that the intact TaF is a heterodimer of two discrete polypeptide chains, one carrying a determinant detectable by a monoclonal anti-Tindd directed to an Igh-I -linked allotypic structure of T cells and being associated with the antigen-binding site and the other expressing a unique determinant controlled by the I-A subregion of murine H-2 major histocompatibility complex but being different from known class II polypeptide chains. The antigen-binding polypeptide has an isoelectric point of pH 5.6, and the I-A polypeptide has an isoelectric point of pH 6.3.

Aged↗

Unique T cell Ia antigen expressed on a hybrid cell line producing antigen-specific augmenting T cell factor.

Hybrid cell lines were established by fusion between keyhole limpet hemocyanin(KLH) binding T cells of A/J mice and an AKR T cell tumor line, BW5147. Hybrids were selected for the presence of Ia antigen and KLH-specific augmenting activity of their extracts in the secondary antibody response. The detailed phenotypic and functional analysis of 1 of these clones, FL10, is reported here. The hybrid was positive for both Thy1.1 and Thy1.2 antigens and possessed the Lyt-1+,2-,3- phenotype. Both VH and Ia determinants were detected on their cell surface. The IA locus was mapped in the I-A subregion, but the Ia specificities were serologically distinct from those of B cell Ia antigen. This was demonstrated by the fact that anti-Ia antiserum preabsorbed with B cells could react with the hybrid cells, whereas none of the monoclonal anti-Ia specific for private and public determinations of Iak could. The extract from the cell line specifically augmented the in vitro secondary antibody response against dinitrophenylated KLH, and this activity was removed by absorption with antigen and conventional anti-Ia antisera. The results indicate that the cell line, FL10, carries Ia antigen unique to the T cell, which is associated with the antigen-specific augmenting molecule.

Absorption↗

Photochemical studies of 7-cis-rhodopsin at low temperatures. Nature and properties of the bathointermediate.

The photoreaction of 7-cis-rhodopsin derived from 7-cis-retinal and cattle opsin was studied by low-temperature spectrophotometry. Upon irradiation of 7-cis-rhodopsin at liquid nitrogen temperature (-190 degrees C) with blue light, its spectrum shifted to the longer wavelengths, indicating the formation of a bathoproduct. The bathoproduct thus formed was found to be identical with bathorhodopsin formed from rhodopsin in their spectroscopic, photochemical, and thermal properties. Therefore, we believe that the bathoproduct is, in fact, bathorhodopsin. The fact that 7-cis-rhodopsin can be readily converted to rhodopsin and to 9-cis-rhodopsin shows that the identical retinal binding site of opsin is involved in the three isomeric rhodopsins. These results appear to be consistent with the notion that the chromophore of bathorhodopsin is a twisted all-trans isomer, which is readily obtainable from the 7-cis, 9-cis, and 11-cis isomers.

Animals↗

Requirement and utilization of egg protein by Japanese young men with marginal intakes of energy.

The effect of marginal intakes of energy on the requirement and utilization of egg protein was evaluated in 46 Japanese young men. The subjects were given a standard diet for 1 week and then low protein diets for 2 weeks. These diets contained about 32, 64, and 80 mgN/kg with whole eggs as the protein source. In the first experiment with excess energy, the energy intakes of 31 subjects were kept constant during the 3 week experiment, the mean intakes being 48.2 +/- 1.5 kcal/kg. The body weight was affected by changing protein intakes while maintaining energy intakes at 48 kcal/kg. From regression analysis, the N requirement for apparent N equilibrium was estimated to be 82.0 +/- 8.0 mgN/kg, where NPU was calculated as 56. In the second experiment with submaintenance energy, 15 subjects received 40 kcal/kg. The N requirement was 124 +/- 21 mgN/kg, where NPU was calculated as 37. From these results and those of previous studies, it was concluded as follows: 1) N balance and NPU were remarkably affected by energy intake changed around maintenance level; and 2) the NPU for egg protein in young men for maintenance intakes of energy and N is about 50 to 55. For estimation of the protein requirement for Japanese adults, a correction factor of 100/55 (about 1.8) was used instead of 1.3 adopted by the 1973 FAO/WHO.

Adult↗

Nutritive efficiencies of lactalbumin and wheat gluten at very low levels of intake in adult rats.

The nutritive values of proteins in relation to their intake levels were evaluated by feeding adult male rats weighing 250 g diets containing 0%, 0.39%, 0.78%, 1.56%, 2.34%, 3.90%, 7.79% and 15.58% lactalbumin or wheat gluten for three weeks. The biological values (BV) of both proteins were high at low levels of protein intake but decreased with increase in protein intake. The BV of wheat gluten was estimated to be about 100 at a level of intake of 1.56% but only 25 at a level of 15.58%. Similarly, the BV of lactalbumin decreased with increase in the protein level, being 67 at a level of 7.79%. The BVs of both proteins at low levels of dietary protein (below 2.34% of lactalbumin or 0.78% of wheat gluten) were apparently more than 100 because urinary N excretion was less than endogenous N. The BVs also decreased with time during the three-week test period. It is concluded that BV of a protein is not a fixed value but varies with the experimental conditions especially with changes in the amount of intake, and that differences in the qualities of various proteins cannot be compared quantitatively at a single level of protein. The results were briefly discussed in relation to protein requirements.

Age Factors↗