Biomedical subjects

H Daniel

Publications and source records attributed to H Daniel.

At least 109 records · Page 6Linked to original sources

PubMed1993

Dipeptides in parenteral nutrition: from basic science to clinical applications.

The use of intravenous dipeptides shows great promise as an avenue for the provision of amino acids that may otherwise be difficult to deliver via nutrient infusions. The physical/chemical properties and metabolism of numerous dipeptides have now been explored in experimental and human studies. It has been found that these agents have the capacity to spare nitrogen and support serum protein levels in a fashion equivalent to that of intravenous free amino acids. An additional benefit is the ability to deliver certain amino acids that are relatively unstable or poorly soluble in aqueous solutions. These various aspects of intravenous dipeptides are considered in this review.

Animals↗

PubMed1992

Determinants of substrate affinity for the oligopeptide/H+ symporter in the renal brush border membrane.

We and others have shown previously the existence of high and low affinity systems for oligopeptide transport in kidney brush border membrane vesicles (BBMV). In the present study we investigated the relationship between the structure of substrates and their affinity for interaction with the high-affinity oligopeptide/H+ transporter in kidney BBMV. Based on competition experiments using [3H]Gly-Gln as a probe we determined the Ki values for more than 60 selected peptides. For a high-affinity interaction with the carrier site the following structural features of substrates are required: (a) both a free amino and carboxyl terminus; (b) the amino group and peptide bond nitrogen located in the alpha-position; (c) a trans peptide bond rather than the cis configuration; (d) L-alpha-amino acid isomers in both COOH and NH2 termini, although D-isomers of hydrophobic amino acids are acceptable in the NH2 terminus; and (e) a backbone of less than 3 amino acid residues. A striking finding of the present study is that, for peptides satisfying these minimal structural requirements, the primary determinant of affinity is hydrophobicity. The fact that there is a highly significant (p less than 0.001) correlation between Ki and hydrophobicity allows the prediction of the affinity for any di- or tripeptide composed of alpha-amino acids in the L-form.

Amino Acid Sequence↗

PubMed1992

Cerebellar nuclei and the nucleocortical projections in the rat: retrograde tracing coupled to GABA and glutamate immunohistochemistry.

The amino acids GABA and glutamate (Glu) are thought to be the principal substances in the central nervous system responsible for neuronal inhibition and excitation. Their distributions among the different neurons in a defined pathway may thus be indicative of the contributions of the cells to pathway function. Examples of such neurons are those of the cerebellar nuclei which, while regulating output from the Purkinje cells of the cerebellar cortex, are also found to project back to the cerebellar cortex. Immunohistochemical experiments were done to identify GABA and glutamate (Glu) containing cells in the adult rat cerebellar nuclei. Consecutive semithin and serial vibratome sections were incubated with antisera raised in rabbit against GABA and Glu. In semithin sections, only small neurons were intensely GABA immunoreactive (GABA-IR) (31.7%), and the majority (80.5%) were Glu immunoreactive (Glu-IR) of different sizes. Consistent with Glu being a metabolic precursor for GABA, 75.4% of the GABA-IR population colocalized Glu. In vibratome sections GABA-IR neurons showed some local differences in number, whereas the Glu-IR were uniformly distributed in the three nuclei studied. Measured mean diameters for these neurons showed a distinct size difference for the GABA- and Glu-IR with little overlap. Cerebellar nuclei neurons projecting to the cortex (nucleocortical neurons, NCN) were identified by locally preinjecting the retrograde transported WGA-apoHRP-colloidal gold complex in the cerebellar cortex. Vibratome sections of these cerebellar were silver intensified for the retrograde tracer and double labeled for GABA and Glu. Of the total number of identified NCN, 8.7% were GABA-IR (10 animals) and 47.7% Glu-IR (5 animals). Many retrograde labeled NCN in the core of the thick sections were immunonegative for both amino acids due to poor antibody penetration, thus underestimating the proportions of cells containing GABA and Glu. The size distributions for the GABA-IR and Glu-IR NCN were similar to those measured in non-retrograde labeled nuclei in thick sections. The conclusions reached are that GABA-IR neurons of the cerebellar nuclei, including the NCN, use GABA as the presumed inhibitory neurotransmitter and that Glu-IR neurons may use Glu or another excitatory neurotransmitter.

Afferent Pathways↗

PubMed1992

Coactivation of metabotropic glutamate receptors and of voltage-gated calcium channels induces long-term depression in cerebellar Purkinje cells in vitro.

Using an in vitro slice preparation, we studied the effects, on parallel fiber (PF)-mediated EPSPs, of coactivation of metabotropic-glutamate receptors and of voltage-gated calcium (Ca) channels of Purkinje cells (PCs) by bath application of 50 microM trans-1-amino-cyclopentyl-1,3-dicarboxylate (trans-ACPD) and by direct depolarization of the cells, respectively. These effects were compared with changes in synaptic efficacy obtained when alpha-amino-3hydroxy-5-methylisoxalone-4-propionate (AMPA) receptors of PCs were also activated through stimulation of PFs during the pairing protocol, as well as when similar experiments were performed without trans-ACPD in the bath. In a control medium, pairing for 1 min of PF-mediated EPSPs evoked at 1 Hz with Ca spikes evoked by steady depolarization of PCs (n = 13) led to LTD of synaptic transmission in 9 cases whereas for the others EPSPs were not affected. No LTD occurred in 9 out of 10 other cells tested when PF stimulation was omitted during the 1 min period of Ca spike firing of PCs. Bath application of 50 microM trans-ACPD, in conjunction with the same pairing protocol as before (n = 8), led to a significantly larger LTD of PF-mediated EPSPs after washing out of this drug. Moreover, a clear-cut LTD of PF-mediated EPSPs was also observed in 5 of the 8 other cells, when PF stimulation was omitted during Ca spike firing in the presence of trans-ACPD.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1992

Sodium-dependent transport of riboflavin in brush border membrane vesicles of rat small intestine is an electrogenic process.

Transport of riboflavin across the intestinal brush border membrane in rats was studied using the brush border membrane vesicle technique. Uptake of riboflavin into the membrane vesicles was predominantly the result of transport into an osmotically reactive space. Binding of the substrate to the membrane surface increased as a function of time and with lowering of the incubation buffer pH. The transport of riboflavin at low substrate concentrations was carrier mediated and Na+, but not K+, dependent with a distinct overshoot phenomenon. Uptake as a function of substrate concentration revealed dual transport characteristics in the presence of a Na+ gradient but linearity in the presence of a K+ gradient. The apparent Km value of the saturable, carrier-mediated and Na(+)-dependent component was calculated by two independent methods to be 0.25 or 0.38 mumol/L. Because uptake of riboflavin into brush border membrane vesicles in the presence of a Na+ gradient was increased by a valinomycin-induced K(+)-diffusion potential and by sodium thiocyanate vs. NaCl (intravesicular negative membrane potential), riboflavin transport is most likely an electrogenic process. The Na(+)-dependent riboflavin uptake into the vesicles was inhibited by structural analogues at low substrate concentrations only and increased by a low buffer pH.

Animals↗

PubMed1992

Oligopeptides: mechanism of renal clearance depends on molecular structure.

We have investigated the relative contribution of hydrolysis, intact transport and urinary excretion to the renal clearance of Gly-Sar, Gly-Sar-Sar, and Gly-Gly-Sar in fed and starved rats. The results obtained from isolated kidney perfusion studies are summarized as follows: 1) clearance was fastest for Gly-Gly-Sar and slowest for Gly-Sar-Sar, 2) urinary excretion of Gly-Sar-Sar exceeded that of Gly-Gly-Sar or Gly-Sar, 3) there was accumulation of products of hydrolysis of Gly-Gly-Sar in the perfusate but not of Gly-Sar or Gly-Sar-Sar, 4) isolated brush-border and basolateral membranes of renal tubular cells lacked hydrolytic activity against Gly-Sar and Gly-Sar-Sar but possessed hydrolytic activity against Gly-Gly-Sar, 5) an excess amount of Gly-Sar-Sar reduced the rate of clearance of Gly-Gly-Sar by approximately 40% and significantly increased urinary excretion of this peptide, 6) the nonfiltering kidney cleared Gly-Gly-Sar at a rate which was 50% of that of the filtering kidney but did not clear Gly-Sar, and 7) starvation for 96 h was without a significant effect on the renal clearance of either Gly-Sar or Gly-Sar-Sar but significantly reduced the renal clearance of Gly-Gly-Sar and the brush-border membrane hydrolase activity against this peptide. We conclude that the molecular structure determines the affinity of oligopeptides for the membrane transport and hydrolytic systems, which, in turn, determines their efficiency for clearance by the kidney.

Animals↗

PubMed1991

Sensitivity of rubrospinal neurons to excitatory amino acids in the rat red nucleus in vivo.

Responses of rubrospinal neurons (RSNs) to iontophoretic applications of L-glutamate (L-Glu), L-aspartate (L-Asp), quisqualate (Quis) and N-methyl-D-aspartate (NMDA) have been studied in the rat red nucleus (RN) in vivo. All agonists produced a dose-dependent increase of the firing rate and Quis was found to be the most efficient. The responses to NMDA and to a lesser extent to L-Asp were abolished by steady application of 2-amino-5-phosphonovalerate (2APV) whereas responses to Quis were unaffected and those to L-Glu poorly antagonized. On the other hand, NMDA-mediated excitations were insensitive to steady application of 6,7-dinitroquinoxaline-2,3-dione (DNQX) which abolished responses to Quis and to a lesser extent to L-Glu while those to L-Asp were less affected. These results show the presence of both NMDA and non-NMDA receptors on RSNs in the rat. A specific localization of the NMDA receptors on distal dendrites of these neurons is suggested.

2-Amino-5-phosphonovalerate↗

PubMed1991

The high and low affinity transport systems for dipeptides in kidney brush border membrane respond differently to alterations in pH gradient and membrane potential.

The principal aim of the present study was to investigate the effects of variation in proton gradient and membrane potential on the transport of glycyl-L-glutamine (Gly-Gln) by renal brush border membrane vesicles. Under our conditions of transport assay, Gly-Gln was taken up by brush border membrane vesicles almost entirely as intact dipeptide. This uptake was mediated by two transporters shared by other dipeptides and characterized as the high affinity (Kt = 44.1 +/- 11.2 microM)/low capacity (Vmax = 0.41 +/- 0.03 nmol/mg protein/5 s) and low affinity (Kt = 2.62 +/- 0.50 mM)/high capacity (Vmax 4.04 +/- 0.80 nmol/mg protein/5 s) transporters. In the absence of a pH gradient, only the low affinity system was operational, but with a reduced transport capacity. Imposing a pH gradient of 1.6 pH units increased the Vmax of both transporters. Kinetic analysis of the rates of Gly-Gln uptake as a function of external pH revealed Hill coefficients of close or equal to 1, indicating that transporters contain only one binding site for the interaction with external H+. The effects of membrane potential on Gly-Gln uptake were investigated with valinomycin-induced K+ diffusion potentials. The velocity of the high affinity system but not of the low affinity system increased linearly with increasing inside-negative K+ diffusion potentials (p less than 0.01). The Kt of neither system was affected by alterations in either pH gradient or membrane potential. We conclude that (a) the high affinity transporter is far more sensitive to changes in proton gradient and membrane potential than the low affinity transporter and (b) in the presence of a pH gradient, transport of each dipeptide molecule requires cotransport of one hydrogen ion to serve as the driving force.

Animals↗

PubMed1991

Effects of ACPD and AP3 on parallel-fibre-mediated EPSPs of Purkinje cells in cerebellar slices in vitro.

The effects of trans-1-amino-cyclopentyl-1,3-dicarboxylate (trans-ACPD) and of DL-2-amino-3-phosphonopropionic acid (AP3), i.e. selective agonist and antagonist of metabotropic quisqualate receptors respectively, on parallel fibre (PF)-mediated EPSPs of Purkinje cells (PCs) were studied in an in vitro slice preparation. Bath application of 500 microM trans-ACPD in conjunction with PF stimulation at 0.2 or 1 Hz depending on the cell always induced a marked depression of PF-mediated EPSPs, which was fully reversible in most cases after wash-out of this compound. Trans-ACPD also often induced a transient depolarization of PCs which induced calcium spike firing in these cells and which again no longer persisted after wash-out of trans-ACPD. Even in cells which were depolarized by trans-ACPD, the decrease in amplitude of PF-mediated EPSPs started before the appearance of calcium spikes, lasted longer than the transient depolarizing effect of trans-ACPD, and was accompanied by no variation in input resistance of the cells when they were manually clamped at their initial resting potential. Bath application of 600 microM DL-AP3 had no effect on PF-mediated EPSPs or the bioelectrical activities of PCs. Moreover, it did not prevent the effects of trans-ACPD mentioned before. The present results are not consistent with the view that coactivation of ionotropic and metabotropic quisqualate receptors of PCs is sufficient to induce a long-term depression of PF-mediated EPSPs.

Alanine↗

PubMed1991

Transport of pteroylglutamic acid into brush border membrane vesicles from rat small intestine is a partially carrier-mediated process.

Intestinal transport of PteGlu was studied using BBMV from rat small intestine. Transport was neither coupled to a specific cation gradient nor was it influenced by variations of the membrane potential. In the presence of a transmembrane pH gradient (pHout less than pHin) initial transport was significantly higher compared to studies without pH gradient. Under these conditions transport could be inhibited by pretreating the vesicles with DIDS, an inhibitor of anion exchange systems. Uptake of PteGlu could not be enhanced by preloading the BBMV with HOP4(2-) and Cl- and was not sensitive to DIDS under these conditions. Uptake studies using different concentrations of PteGlu revealed dual transport kinetics in the presence of a pH gradient and linear uptake in its absence. It could be concluded that uptake is mediated by a PteGlu-/OH(-)-antiporter at low substrate concentrations and occurs by non-ionic diffusion at higher concentrations or in the absence of a pH gradient. In an additional series of experiments it could be shown that about one-third of the substrate is bound to the membrane and is not transported. The biological significance of this binding remains unclear.

Animals↗

PubMed1991

In vivo kinetics of intestinal absorption of riboflavin in rats.

To investigate absorption kinetics of riboflavin under in vivo conditions, with blood and lymph circulation intact, the small intestine of anesthetized rats was perfused with [14C]riboflavin in a concentration range between 0.31 and 10.00 mumol/L. Apart from the uptake of riboflavin from the perfusate, passage of the vitamin into the portal (vena portae) and peripheral (vena femoralis) blood was determined. The absorption proved to be a dual process: at low substrate concentrations (less than 2 mumol/L) a saturable component predominated; at higher concentrations simple diffusion was found to be the prevailing uptake mechanism. The apparent transport constant of the saturable component was calculated to be 0.38 mumol/L. [14C]flavin concentrations in the portal and peripheral blood were estimated as a function of the riboflavin concentration of the perfusion media. The dual character of the absorption was reflected by the portal blood flavin levels. Due to the high retaining and equalizing capacity of the liver, the [14C]flavin level of the peripheral blood was relatively low and obeyed saturation kinetics. Constants of elimination, determined by pharmacokinetic calculations, were different for the two blood compartments but independent of the concentration of riboflavin in the perfusion media.

Animals↗

PubMed1991

We are all people living with AIDS: myths and realities of AIDS in Brazil.

Although AIDS was expected in Brazil, no serious efforts were undertaken to prevent AIDS from taking root. Irresponsible press and media coverage highlighted the spread of AIDS within the gay community of the United States, creating an aura of immunity in Brazil to what was characterized as a "foreign" disorder. When AIDS did surface in 1983, the official response was to adopt an abstract, inappropriate, and ideological "Western" model, in which only stigmatized "others" and "minorities" were at risk of HIV infection. Brazilian health authorities subsequently downplayed the significance of the sale of contaminated blood in HIV transmission, and likewise ignored the rising rates of AIDS among Brazil's one unarguable majority group: the poor. An analysis of efforts to force the "facts" of AIDS to fit a false model's predictions leads to a clearer definition of the broader context of the Brazilian epidemic: we all are people living with AIDS, precisely because we live in this age of AIDS; it is sheer folly to discriminate against persons infected by HIV and to obstruct their participation in efforts to curtail the epidemic's spread; and the necessary response to AIDS is solidarity, not because it is poetic, but because no other response will suffice.

Acquired Immunodeficiency Syndrome↗

PubMed1991

Improved xenograft survival with continuous infusion deoxyspergualin and RATG.

The critical shortage of available donor organs is the major limit to current allogeneic transplantation. Xenografting has the potential to overcome this difficult problem. Suppressing the vigorous rejection response remains a major obstacle to the clinical application of xenografting, 15-Deoxyspergualin (DOSP), a potent new immunosuppressive agent has been shown to be effective in allogeneic and xenogeneic experimental models. This study tests DOSP in combination with rabbit antithymocyte globulin (RATG) in the hamster-to-rat cardiac xenograft. Results show that combination therapy with DOSP/RATG was superior to treatment with either agent alone (p less than .05). Optimal graft prolongation (20.9 days versus control of 3.1 days, p less than .05) was achieved with combination therapy of RATG and DOSP 2.5 mg/kg day-1 by continuous infusion.

Animals↗

PubMed1990

Effect of casein and beta-casomorphins on gastrointestinal motility in rats.

The effect of bovine casein and synthetic beta-casomorphins on the motility of rat gastrointestinal tract was studied by noninvasive techniques using the nonabsorbable marker 141Ce. Casein suspensions (CAS) or whey protein suspensions (WPS) were labeled with 141Ce and fed by gastric tube. Gastric emptying rate (GER) as well as gastrointestinal transit time (GITT) of the tracer were significantly longer with feeding CAS compared to WPS. The differences between the CAS and the WPS groups were partly (GER) or completely (GITT) abolished by pretreating the animals with the specific opiate-receptor antagonist naloxone. It is assumed that opioid peptides released from casein during digestion slowed gastrointestinal motility by direct interaction with gut opiate receptors. To prove whether beta-casomorphins, when given by gastric tube, can affect motility, different synthetic beta-casomorphins in doses between 1 and 10 mg were added to the WPS. The beta-casomorphin-4 (Tyr-Pro-Phe-Pro-NH2) showed no effect on GITT. The D-Ala substituted D-Ala-beta-casomorphin-4 (Tyr-D-Ala-Phe-Pro-NH2) and D-Ala-beta-casomorphin-5 (Tyr-D-Ala-Phe-D-Ala-Tyr-NH2), which are more resistant to proteolytic attack and have higher opioid potency than beta-casomorphin-4, slowed GITT in a dose-dependent manner.

Amino Acid Sequence↗

PubMed1990

New tetracyclic guanidine derivatives with H1-antihistaminic properties. Chemistry of epinastine.

A series of new tetracyclic guanidines were synthesized by various methods. Specific binding of the described compounds to histamine-1 and histamine-2 receptors was determined. The compound 3-amino-9,13b-dihydro-1H-dibenz[c,flimidazo[1,5-a]azepine (epinastine, WAL 801, compound IIIa) combines high selectivity with high affinity for the H1 receptor and was selected from the compounds studied for further pharmacological and clinical investigations. Experimentally determined physicochemical parameters (pka-value, partition coefficient) and the hydrogen-bonding ability of epinastine are indications that this compound will not easily cross the blood-brain barrier. This explains the absence of CNS side-effects of epinastine in pharmacological and clinical studies.

Animals↗

PubMed1989

The GABAergic neurones of the cerebellar nuclei in the rat: projections to the cerebellar cortex.

The presence of gamma-aminobutyric acid (GABA) in the neurones of the cerebellar nucleocortical pathway is here reported. The pathway was identified by retrograde tracer and the GABA content was revealed immunohistochemically. It was found that most of the neurones giving rise to the reciprocal, non-reciprocal and symmetrical projections are indeed GABA-immunoreactive. They were observed in all the subdivisions of the nucleus medialis, of the nucleus interpositus and of the nucleus lateralis sending axons respectively to the sagittal zones A, C1-3 and D of the cerebellar cortex. The nucleus vestibularis lateralis and the related sagittal zone B were devoid of such projections.

Animals↗

PubMed1989

Demonstration and modification of intervillous pH profiles in rat small intestine in vitro.

The intervillous pH profiles along the crypt villus axis in different regions of the rat small intestine were measured in vitro by using pH-sensitive liquid ion-exchanger microelectrodes. A characteristic pH profile was observed in the duodenum and jejunum. A region of low pH was detected in the upper parts of the villi (pH 6.65 +/- 0.06 to 6.85 +/- 0.07), whereas pH at the villus base was always higher. In the ileum no gradient was observed (pH 7.26 +/- 0.05 to 7.31 +/- 0.05). Preincubation of the tissue in situ with 10 mM theophylline for 1 h caused an increase in the villus base pH in the jejunum (pH 7.24 +/- 0.04) and ileum (7.44 +/- 0.04) followed by a subsequent increase of the pH in the upper part of the villi. These results indicate that the low pH in the upper intervillous space may be related to H+ secretion occurring from the mature enterocytes, whereas the crypt cells may secrete a rather neutral or slightly alkaline fluid. Alkaline secretion from the crypts may be increased by theophylline, which changes the levels of cyclic nucleotides in the mucosa.

Animals↗

PubMed1989

Peroxidase activities in the hamster bronchoalveolar lining fluid: modifications induced by exposure to silica dust.

The modifications of peroxidase (Po) activity have been studied in bronchoalveolar lavage fluid (BALF) from hamsters exposed to silica dust. In silica-treated animals, the mean total BALF-Po activity was significantly increased compared to control animals. This increased activity was accompanied by an influx of polymorphonuclear neutrophils in airways. HPLC gel filtration of BALF from control animals separated 5 peaks with Po activity. They had an apparent molecular weight of 140, 110, 80, 57, and 42 kDa. In BALF from silica-exposed animals, with the exception of the 57-kDa fraction, the same peaks were found. Additional fractions with an apparent molecular weight of greater than 200, 180, 92, 65, and 20 kDa were detected. All the fractions but those at 57 and 92 kDa were detectable in a whole-blood homogenate. Exposing hamsters to silica induced both quantitative modifications and a different pattern of BALF proteins having Po activity in the alveolar lining fluid.

Animals↗