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Biomedical subjects

T A Davis

Publications and source records attributed to T A Davis.

At least 73 records · Page 4Linked to original sources
PubMed1996

Effect of spaceflight on human stem cell hematopoiesis: suppression of erythropoiesis and myelopoiesis.

Humans subjected to periods of microgravity develop anemia, thrombocytopenia, and abnormalities in red blood cell structure. The causes of these abnormalities are complex and unclear. The in vitro effects of spaceflight on hematopoietic cell proliferation and differentiation were investigated during the space shuttle missions STS-63 (Discovery) and STS-69 (Endeavour). CD34+ bone marrow progenitor cells were cultured in liquid suspension culture and on hematopoietic supportive stromal cells using hollow-fiber culture modules. One set of cultures was maintained at microgravity (flight cultures) for the last 8-10 days of culture and a second control was at full gravity (ground control). Over the 11- to 13-test-day period, ground control culture total cell number increased 41.0- to 65.5-fold but flight culture total cell number increased only 10.1- to 17.6-fold (57-84% decrease). Comparing ground control cultures and microgravity cultures, respectively, for progenitor cell content, myeloid progenitor cell numbers expanded 2.6- to 17.5-fold compared with 0.9- to 7.0-fold and erythroid progenitor cell numbers expanded 2.0- to 4.1-fold in ground control cultures but actually declined at microgravity (>83% reduction). Moreover, microgravity cultures demonstrated accelerated maturation/differentiation toward the macrophage lineage. These data indicate that spaceflight has a direct effect on hematopoietic progenitor cell proliferation and differentiation and that specific aspects of in vitro hematopoiesis, particularly erythropoiesis, involve gravity-sensitive components.

Antigens, CD↗
PubMed1996

Opposing effects of CTLA4-Ig and anti-CD80 (B7-1) plus anti-CD86 (B7-2) on experimental allergic encephalomyelitis.

The roles of the B7 receptors, CD80 and CD86, during actively induced experimental allergic encephalomyelitis were examined with specific monoclonal antibodies and CTLA4-Ig. Injection of CTLA4-Ig on day 2 post-immunization resulted in decreased incidence and severity of resultant disease. Anti-CD80 injection on day 2 blocked development of the first disease episode. Subsequent relapses were unaffected. In contrast, injection of anti-CD86 alone had no effect. Surprisingly, combined anti-CD80 + anti-CD86 monoclonal antibody injection on day 2 resulted in marked exacerbation of disease. Examination of cytokine production in the draining lymph node cells demonstrated a reduction in both interferon (IFN)-gamma and interleukin (IL)-2 producing cells, but a dramatic increase in tumor necrosis factor (TNF)-alpha secretion in animals receiving both monoclonal antibodies. These results suggest distinct roles for CD80 and CD86 in the initiation of EAE, resulting in the diverse clinical outcomes observed in this model of EAE.

Abatacept↗
PubMed1996

Whole-body protein turnover in the fed state is reduced in response to dietary protein restriction in lactating women.

We examined the adaptive responses of body protein metabolism in the fed state to dietary protein restriction in lactating women to determine whether rates of body protein degradation and synthesis were lower than those of nonlactating women. Thirteen healthy women (five lactating, four nonlactating postpartum, four nulliparous) aged 28-32 y were given protein intakes of 1.5, 0.4, and 1.0 g.kg-1.d-1 over three consecutive 3-d periods, respectively. At the end of each period, while in the fed state, subjects received orally a single bolus dose of [1-13C]leucine. A 24-h urine collection was obtained simultaneously. Whole-body protein metabolism was characterized by using the end product model based on nitrogen excretion and leucine catabolism. Nitrogen flux and rates of protein degradation and synthesis in the fed state were significantly lower at a dietary protein intake of 1.0 g.kg-1.d-1 in lactating women than in their nonlactating postpartum counterparts. Net protein retention in the fed state was significantly higher at a dietary protein intake of 1.0 g.kg-1.d-1 in lactating than in nonlactatating postpartum and nulliparous women because of the relatively greater reduction in protein degradation than in protein synthesis. These studies suggest that lactating women rapidly adapt to dietary protein restriction by down-regulating protein metabolism, and that 13C-labeled amino acid tracers in combination with urinary nitrogen excretion serve as useful metabolic markers for the adequacy of the dietary protein content of lactating women.

Adult↗
PubMed1996

Primary porcine endothelial cells express membrane-bound B7-2 (CD86) and a soluble factor that co-stimulate cyclosporin A-resistant and CD28-dependent human T cell proliferation.

Increasing evidence suggests that endothelial cells can directly activate syngeneic, allogeneic and xenogeneic T cells. In this study we demonstrate that unstimulated, paraformaldehyde-fixed primary porcine aortic endothelial cells (PAEC) and microvascular endothelial cells (PMVEC) can provide co-stimulation for human T cell IL-2 secretion and proliferation. EC-mediated co-stimulation has both cyclosporin A (CsA)-sensitive and CsA-resistant components. The CsA-resistant component is completely suppressed either by blocking with anti-CD28 F(ab) fragments or CTLA-4-Ig. Northern blot analysis of unstimulated PAEC and PMVEC with porcine-specific probes reveals constitutive expression of B7-2 mRNA while B7-1 message was not detected. hCTLA-4-Ig and anti-B7-2 mAb immunoprecipitates a single 79 kDa PMVEC surface protein. Surprisingly, PMVEC conditioned media also has soluble co-stimulatory activity that is blocked by anti-CD28 F(ab) fragments or anti-B7-2 mAb. These findings demonstrate that primary unstimulated porcine EC can co-stimulate CsA-resistant human T cell proliferation through binding of membrane bound, constitutively expressed EC B7-2 (CD86) to human T cell CD28, providing one of the first demonstrations of functional B7-2 on cells outside the immune system. In addition, PMVEC secrete or shed a soluble factor that mediates CD28-dependent human T cell proliferation, demonstrating the existence of soluble mediators of CD28 activation.

Animals↗
PubMed1996

Protein synthesis in skeletal muscle and jejunum is more responsive to feeding in 7-than in 26-day-old pigs.

The study aimed to determine the developmental changes in the response of peripheral and visceral tissue protein synthesis to feeding during early postnatal life and the associated changes in circulating insulin, insulin-like growth factor (IGF-I), and amino acid concentrations. Tissue protein synthesis was measured in vivo with a large dose of L-[4(-3)H]phenylalanine in 7- and 26-day-old pigs that were either fasted for 24 h or refed for 2.75 h after a 24-h fast. Fractional rates of protein synthesis (Ks) in skeletal muscle, heart, and liver were greater in 7-than in 26-day-old pigs. Refeeding stimulated Ks in skeletal muscle, pancreas, jejunum, and liver of both 7-and 26-day-old pigs. The stimulation of skeletal muscle and jejunal Ks by refeeding was greater in 7- than in 26-day-old pigs. Plasma IGF-I concentrations were lower in 7- than in 26-day-old pigs. Plasma concentrations of insulin and amino acids increased with refeeding. The increase in plasma insulin concentrations with refeeding was greater in 7- than in 26-days-old pigs. These results indicate that the stimulation in skeletal muscle and jejunal protein synthesis by feeding is elevated in young compared with older suckling pigs. This enhanced stimulation of protein synthesis by feeding in neonatal pigs is associated with elevated circulating concentrations of insulin but not amino acids or IGF-I.

Aging↗
PubMed1996

Orally administered IGF-I increases intestinal mucosal growth in formula-fed neonatal pigs.

Our objective was to determine the potentially anabolic effects of orally administered recombinant human insulin-link growth factor I (rhIGF-I)on small intestinal growth in formula-fed neonatal pigs. Unsuckled neonatal pigs received formula or formula containing added rhIGF-I (3.5 mg.kg-1.day-1) from birth to 4 days of age. Pigs in both groups were fed 30 ml/kg formula every 2 h on day 1 and then every 4 h on days 2-4, and blood was sampled daily. Oral administration of rhIGF-I to formula-fed neonatal pigs increased small intestinal weight, protein, and DNA content,but not length. Jejunal and ileal villus height, but not crypt depth or muscularis thickness, also were increased by oral rhIGF-I administration. Neither the circulating concentration of IGF-I nor the IGF-binding proteins differed between control and oral rhIGF-treated pigs, suggesting that the absorption of orally administered rhIGF-I from the intestinal lumen into the peripheral circulation was limited. Our results demonstrate that oral administration of rhIGF-I during the first 4 days after birth significantly increased small intestinal mucosal growth in formula-fed neonatal pigs. These results suggest that oral administration of rhIGF-I may be a viable therapeutic approach to enhance intestinal growth in neonates.

Administration, Oral↗
PubMed1996

Corticosterone has independent effects on tissue maturation and growth in the suckling rat.

Glucocorticoids are associated with reduced weight gain when used to improve pulmonary function in premature infants. However, tissue maturation is stimulated during normal development by an increase in serum glucocorticoids. We evaluated the effects of glucocorticoid treatment on tissue weight gain and the activity of specific enzymes in the suckling rat, with the hypothesis that these processes are independently regulated. Before the ontogenic surge in corticosterone, 6-d-old rat pups were implanted with a pellet to release corticosterone continuously at 0 (placebo), 48, 120, 240, or 360 micro g/d. We killed the pups at 7, 9, or 12 d of age and measured tissue weights and activities of sucrase and glutamine synthetase. Serum corticosterone concentrations were elevated with dose. Tissue weight gain was proportional to ln(e) serum corticosterone at all ages. In contrast, enzyme indices of tissue maturation did not respond to corticosterone until d9. Also, intestinal tissue was more sensitive than muscle to the effects of corticosterone on weight but less sensitive to its effects on maturation. We conclude that the immediate response, in terms of weight versus the delayed response of the enzymes and their reciprocal sensitivity in muscle and gut, indicates that these processes are independently regulated.

Animals↗
PubMed1996

Multiple responses to administration of liposome-encapsulated hemoglobin (LEH): Effects on hematopoiesis and serum IL-6 levels.

Liposome-encapsulated hemoglobin (LEH) has been tested in animals as an oxygen-carrying red cell substitute and has been shown to be beneficial in the treatment of hemorrhagic shock. The effects of LEH on immune responses have not been studied thoroughly in any well-controlled model. Using a murine model, we evaluated nephrotoxicity and hepatotoxicity as well as immune function parameters following LEH administration. Following intravenous administration of LEH, 1) a serum spike of interleukin-6 (IL-6) occurred in mice at 4-8 hours, with no elevation of IL-1, tumor necrosis factor (TNF), or interferon-gamma (IFN-gamma); 2) the serum liver function enzymes SGOT (AST, aspartate aminotransferase) and SGPT (ALT, alanine aminotransferase) were elevated at 48 hours; 3) only a slight increase in serum antibody to bovine hemoglobin was observed; and 4) increased hematopoietic activity was observed in the spleen and bone marrow. The finding that only IL-6 but not the associated TNF, IL-1, or IFN-gamma is secreted in vivo following LEH administration is novel and may have significance in defining the mechanisms underlying specific adverse responses observed with LEH administration in animals.

Alanine Transaminase↗
PubMed1995

Porcine brain microvascular endothelial cells support the in vitro expansion of human primitive hematopoietic bone marrow progenitor cells with a high replating potential: requirement for cell-to-cell interactions and colony-stimulating factors.

Primary autologous as well as allogeneic and xenogeneic stroma will support human stem cell proliferation and differentiation for several months. In the present study, we investigated the capacity of porcine microvascular endothelial cells (PMVECs) together with combinations of cytokines (granulocyte-macrophage colony-stimulating factor [GM-CSF] + stem factor [SCF], interleukin-3 [IL-3] + SCF + IL-6, and GM-CSF + IL-3 + SCF + IL-6) to support the expansion and development of purified human CD34+ bone marrow cells. In short-term cultures (7 days), the greatest expansion of nonadherent hematopoietic cells and clonogenic progenitors was seen with CD34+ cells in direct contact with PMVEC monolayers (PMVEC contact), followed by PMVEC noncontact and liquid suspension cultures, respectively. Maximal expansion of nonadherent cells (42-fold) and total CD34+ cells (12.6-fold) occurred in PMVEC contact cultures treated with GM-CSF + IL-3 + SCF + IL-6, with similar increases in the number of granulocyte-macrophage colony-forming units (CFU-GM), CFU-mix, erythroid burst-forming units (BFU-E), CFU-blast and CFU-megakaryocyte (CFU-Mk) progenitor cells. Moreover, the number of CD34+ CD38- and CD34+ CD38+ cells increased 148.1-fold and 8.0-fold, respectively. Replating studies show that cells from day 7 dispersed blast cell colonies generated on cytokine-treated PMVEC monolayers have a high replating potential for multilineage progenitor cells. In long-term PMVEC contact cultures, CD34+ cells seeded onto PMVEC monolayers with GM-CSF + IL-3 + SCF + IL-6 showed a total calculated expansion of over 5,000,000-fold of nonadherent cells over 35 days in culture. Maximal clonogenic cell production was observed at day 28, with 6,353-fold for total CFC and comparable increases for CFU-GM, CFU-mix, CFU-blast, BFU-E, and CFU-Mk. The total number of CD34+ cells increased 2,584-fold at day 28. Furthermore, the extended growth kinetics of these cultures indicates that these phenotypically primitive progenitor cells are also functionally expanded on PMVEC monolayers. These results support the hypothesis that direct contact with a PMVEC monolayer supports the initial expansion of hematopoietic progenitor cells with a high replating potential and, possibly, a more primitive phenotype (CD34+, CD34+/CD38-).

Animals↗
PubMed1995

Both maternal over- and undernutrition during gestation increase the adiposity of young adult progeny in rats.

We examined the influence of maternal diet during gestation on the growth and body composition of the progeny. On day 1 of gestation, rat dams were assigned to one of four feeding regimens: free access to standard rodent chow throughout gestation (AL); 20 g feed/day (prebreeding intake) throughout gestation (PB); 10 g feed/day from day 1 to day 14, then ad libitum from day 15 to parturition (RAL); 10 g feed/day from day 1 to 14, then 20 g/day to parturition (RPB). Progeny were fed ad libitum on standard chow diet from 3 to 12 weeks of age; food intake and weight gain were measured over this time. Body composition was measured at 12 weeks. The PB regimen restricted maternal food intake during the third trimester only; the RAL regimen restricted intake by 50% for two trimesters and produced hyperphagia in the third; the RPB regimen restricted intake by 50% for two trimesters, then intake (per unit body weight) was similar to that of AL dams during the third trimester. Litter size and progeny birth, weaning, and 12-week body weights were similar among the four groups. At 12 weeks of age, PB progeny had the highest body fat (per kg fat-free mass), despite similar feed intake during the 9-week postweaning period. The increased fat was proportionally distributed among intra-abdominal and subcutaneous depots. Progeny of RAL, AL, and RPB dams had similar amounts of body fat, but in RAL progeny more fat was present in intra-abdominal depots. The weights of fat-free mass, gastrointestinal tract and hindlimb skeletal muscles were unaffected by maternal diet. Restriction of maternal feed intake during the third week of gestation had subtle effects on the body composition of young adult progeny that could not be explained on the basis of differences in postweaning voluntary feed intake.

Abdomen↗
PubMed1995

Amino acid composition of pinniped milk.

The total amino acid concentration and the amino acid pattern, i.e. the relative proportion of each amino acid (protein-bound plus free) to the total amino acids, in the milks of the Northern elephant seal, Antarctic fur seal, California sea lion, and Australian sea lion were determined. Total amino acid concentration was 10% (w/v) or greater and did not vary significantly among species. The most abundant amino acids in the milks of all species were glutamate, proline and leucine. Essential amino acids were 40%, branched-chain amino acids were 20%, and sulfur amino acids were 4% of the total milk amino acids in all species. There were differences among the pinnipeds in some of the individual amino acids; the milk of the Northern elephant seal was the most distinct among the pinnipeds with higher histidine, serine and cystine contents and a lower methionine content than that of other pinnipeds. There was little effect of stage of lactation on total amino acid concentration or amino acid pattern in pinniped milk. Comparison of milk from the four pinniped species with that of 14 other mammalian species suggests commonality in milk amino acid pattern despite the wide variation in total amino acid concentration among the species.

Amino Acids↗
PubMed1995

Nutrient-independent and nutrient-dependent factors stimulate protein synthesis in colostrum-fed newborn pigs.

We hypothesized that nonnutrient components, including growth factors, present in colostrum contribute to the stimulation of protein synthesis in colostrum-fed neonatal pigs. We studied neonatal pigs fed mature milk, colostrum, or a formula containing a macronutrient composition comparable to that of colostrum for 24 h. We measured the circulating concentrations of insulin, insulin-like growth factor I, glucose, and amino acids at intervals throughout the 24-h period, after which we measured in vivo protein synthesis using a flooding dose of [3H]phenylalanine. The rates of protein synthesis in several tissues measured after 24 h of feeding were greater than those we reported previously after 6 h of feeding. The acute (within 6 h) stimulation of protein synthesis in visceral and skeletal muscle tissues of neonatal pigs fed milk, colostrum, or formula was primarily influenced by nutrient intake and associated with rapid secretion of insulin. Indirect evidence suggests that intestinal absorption of ingested colostral insulin was minimal. However, the sustained increase in tissue protein synthesis between 6 and 24 h coincided with an increase in circulating insulin-like growth factor I. We found a novel, specific stimulation of skeletal muscle and jejunal protein synthesis in colostrum-fed pigs that can be attributed to some nonnutrient component of colostrum.

Amino Acids↗
PubMed1994

A protein complex required for signal-sequence-specific sorting and translocation.

We have purified a nascent-polypeptide-associated complex (NAC) which prevents short ribosome-associated nascent polypeptides from inappropriate interactions with proteins in the cytosol. NAC binds nascent-polypeptide domains emerging from ribosomes unless a signal peptide is fully exposed. Depletion of cytosolic proteins (including NAC) from ribosomes carrying nascent polypeptides allows the signal recognition particle (SRP) to crosslink to polypeptides irrespective of whether or not they contain signal peptides. In the absence of cytosol, proteins lacking signal peptides can be mistranslocated into the endoplasmic reticulum in vitro, albeit with low efficiency. Readdition of NAC restores the specificity of SRP and fidelity of translocation.

Amino Acid Sequence↗
PubMed1994

Amino acid composition of the milk of some mammalian species changes with stage of lactation.

To determine whether the amino acid composition of milk changes during lactation, we compared the amino acid pattern (concentration of each individual amino acid relative to the total amino acid concentration) of colostrum with that of mature milk in six mammalian species. In the human, horse, pig and cow, the pattern of amino acids changed between colostrum and mature milk: glutamate, proline, methionine, isoleucine and lysine increased; cystine, glycine, serine, threonine and alanine decreased. In these four species, the total amino acid concentration also decreased 75% between colostrum and mature milk. In the baboon (Papio cynocephalus anubis and Papio cynocephalus anubis/Papio cynocephalus cynocephalus) and rhesus monkey (Macaca mulatta), however, there was little change in the pattern of amino acids between colostrum and mature milk, and total amino acid concentration decreased only about 25% between colostrum and mature milk. Mature milk rather than colostrum was the most similar among the three primates in both amino acid pattern and total amino acid concentration. We conclude, in those species in which total amino acid concentrations decline substantially between colostrum and mature milk, amino acid patterns also change. The presence of a change in amino acid pattern and total amino acid concentration during lactation appears to be unrelated to phylogenetic order.

Amino Acids↗
PubMed1994

Leucine oxidation changes rapidly after dietary protein intake is altered in adult women but lysine flux is unchanged as is lysine incorporation into VLDL-apolipoprotein B-100.

We measured the 13C enrichments of expired CO2 and deuterium enrichments of plasma free lysine and VLDL-apolipoprotein B-100 in five nulliparous women who received an oral bolus dose of [1-13C]leucine and a primed, constant infusion of [2H4]lysine on d 2 and 6 while consuming protein diets of 1.5, 1.0 and 0.4 g.kg-1 x d-1. Peak enrichments and cumulative percent recoveries of 13C in expired CO2 increased during the high, and decreased during the low protein periods within 24 h of altered intakes; these changes averaged 89% of that on d 6 of the high and low protein diets. The early changes in leucine oxidation showed significant relationships with urinary nitrogen excretion on d 6 of the dietary periods. The ratio of the isotopic enrichment of lysine in VLDL-apolipoprotein B-100 to that in plasma was unaltered by the level of protein intake. Thus, amino acid oxidation adapts rapidly to altered protein intakes in adult women and is useful to determine protein needs during rapidly changing physiologic conditions. However, 6-d periods of protein intake over the range of 1.5 to 0.4 g.kg-1 x d-1 do not affect the proportional use of dietary amino acids for hepatic secretory protein synthesis.

Administration, Oral↗
PubMed1994

Amino acid composition of human milk is not unique.

To determine whether the amino acid pattern of human milk is unique, we compared the amino acid pattern of human milk with the amino acid patterns of the milks of great apes (chimpanzee and gorilla), lower primates (baboon and rhesus monkey) and nonprimates (cow, goat, sheep, llama, pig, horse, elephant, cat and rat). Amino acid pattern was defined as the relative proportion of each amino acid (protein-bound plus free) (in mg) to the total amino acids (in g). Total amino acid concentration was lower in primate milk than in nonprimate milk. There were commonalities in the overall amino acid pattern of the milks of all species sampled; the most abundant amino acids were glutamate (plus glutamine, 20%), proline (10%) and leucine (10%). Essential amino acids were 40%, branched-chain amino acids 20%, and sulfur amino acids 4% of the total amino acids. The amino acid pattern of human milk was more similar to those of great apes than to those of lower primates. For example, cystine was higher and methionine was lower in primate milks than in nonprimate milks, and in great ape and human milks than in lower primate milks. Because the milk amino acid patterns of the human and elephant, both slow-growing species, were dissimilar, the amino acid pattern of human milk seems unrelated to growth rate.

Amino Acids↗
PubMed1993

Control of folding and membrane translocation by binding of the chaperone DnaJ to nascent polypeptides.

Recent evidence supports the view that cellular protein folding may be mediated by molecular chaperones. A fundamental question concerns the stage in its biogenesis at which the folding protein makes first contact with these components. We show here by crosslinking that the chaperone DnaJ binds nascent ribosome-bound polypeptide chains as short as 55 residues. Cotranslational binding of DnaJ to firefly luciferase and chloramphenicol acetyltransferase resulted in an arrest of folding as long as the functional partners of DnaJ in Escherichia coli, DnaK and GrpE, were missing. Protein uptake into microsomes and mitochondria was also interrupted by DnaJ. Both folding and post-translational translocation recommenced upon addition of DnaK and GrpE. We propose that DnaJ protects nascent polypeptide chains against aggregation and, in cooperation with Hsp70, controls their productive folding once a complete polypeptide or a polypeptide domain has been synthesized.

Animals↗