Biomedical subjects

B J Van de Heijning

Publications and source records attributed to B J Van de Heijning.

9 recordsLinked to original sources

PubMed1999

Determination of actin content in gallbladder tissue and its relevance to normalization of tensiometry data.

OBJECTIVES: To compare the common procedure in tensiometry of normalization of the force (in N) produced by a gallbladder tissue strip to units of stress, with normalization of force to the strip content of contractile protein. DESIGN: A comparison was made in both healthy and in diseased gallbladder tissue strips between two normalization procedures involving anatomical parameters. The contractile response expressed in terms of tissue stress (in N/m2), which entails a normalization to the strip cross-sectional area, was set against normalization to the tissue content of contractile protein (in N/mg actin/g strip wet weight). METHOD: Dose-response curves for acetylcholine (ACh) (10(-8) to 10(-3) M) and sulphated cholecystokinin octapeptide (CCK) (10(-12) to 10(-6) M) were assessed in healthy guinea pig (n = 8) and in diseased human gallbladder tissue strips (n = 28). Assuming a tissue density of 1.05 g/cm3, the strip cross-sectional area was calculated from its weight and length. Actin content in homogenized strips was determined by polyacrylamide gel electrophoresis followed by densitometry. RESULTS: Actin content in human tissue was 19.06 +/- 1.42 mg/g strip wet weight, and 12.84 +/- 0.76 mg/g strip wet weight in guinea pig tissue. No correlation was found between strip cross-sectional area and actin content. In the diseased human tissue, no correlation was found between the inflammation score and either strip cross-sectional area or strip actin content. Maximal force (in mN) exerted in response to either ACh or CCK correlated much more closely in healthy guinea pig gallbladder (r = 0.97) than in diseased human tissue (r = 0.59). Normalization of maximal force to strip cross-sectional area (i.e. to stress) showed considerable more variation (% coefficient of variance) than the normalization to strip actin content in healthy guinea pig tissue, although both strip cross-sectional area and actin content per se showed little variation. Normalization to either parameter did not result in an improved correlation or a decreased variation in the case of diseased human gallbladder tissue. CONCLUSION: Normalization of muscle strip force in diseased tissue is questionable, as the assumptions made for healthy tissue are not valid.

Acetylcholine↗

PubMed1996

Effects of bile salt hydrophobicity on crystallization of cholesterol in model bile.

Precipitation of cholesterol crystals is an essential step in gallstone formation. In the present study we found much faster and more extensive precipitation of various cholesterol crystal shapes in whole model biles containing the hydrophobic bile salt taurodeoxycholate than in biles containing the relatively hydrophilic taurocholate. Addition of taurodeoxycholate to isolated cholesterol-phospholipid vesicles also induced more crystallization than taurocholate. Crystallization behaviour in whole model biles and in vesicles after addition of corresponding bile salts was very similar. The very hydrophilic bile salts tauroursodeoxycholate and taurohyodeoxycholate never induced crystallization from vesicles, and crystallization in corresponding whole model biles did not occur. These bile salts also reduced crystallization dose dependently after addition of taurodeoxycholate to vesicles. Ultracentrifugation experiments suggested a higher vesicular cholesterol-phospholipid bile salts. These findings indicate that bile salt hydrophobicity influences shape of cholesterol crystals and extent of crystallization, possibly by modulating the vesicular cholesterol-phospholipid ratio.

Bile Acids and Salts↗

PubMed1996

A novel vesicular assay to study factors affecting cholesterol crystallization in vitro.

We present an assay to study cholesterol crystallization that is fast, facile, and highly reproducible. Cholesterol crystallization from metastable vesicles was induced upon addition of bile salts and depended on the hydrophobicity of the bile salt used and the cholesterol-to-phospholipid ratio of the vesicles. Bile salt-induced crystallization was stimulated upon addition of Con A-binding glycoproteins (CABGs), comparable to the results of the same CABGs in a crystal growth assay. The onset time and total measuring time, however, were much shorter. This assay might, moreover, provide a tool to study the mechanism of cholesterol crystallization in more detail.

Bile↗

PubMed1995

Cholecystokinin evokes vasopressin release from perfused hypothalamic-neurohypophyseal explants.

Cholecystokinin (CCK) may have a transmitter/modulator role in the hypothalamic magnocellular neurosecretory system. In the rat, the supraoptic and paraventricular nuclei display high affinity binding for radiolabelled CCK. Exogenously applied CCK depolarizes supraoptic neurons, acting at postsynaptic CCK-B type receptors. The present study evaluated the ability for the sulfated octapeptide of CCK (CCK-8S), which is a predominate form of this peptide in brain, to evoke release of vasopressin from the neurohypophysis of intra-arterially perfused hypothalamic explants. 3 min applications of 1 microM CCK-8S through the intra-arterial perfusion medium prompted an elevation of vasopressin in samples taken from the neurointermediate lobe in 10 of 14 preparations. Vasopressin levels rose from undetectable baseline values to a peak of 29.5 +/- 6.7 pg/ml (mean +/- S.E.M). This response was dose-dependent and was abolished by pituitary stalk transection (5/5 explants). Locally applied CCK-8S (25-200 pmol) through bilateral infusions onto the ventral surface of the supraoptic nucleus also induced a dose-dependent release of vasopressin (5/7 explants). These observations suggest that CCK can act at receptors located on (or near) the somata of supraoptic nucleus neurons to induce neuronal discharges that are conducted to the neural lobe where they evoke release of vasopressin from neurohypophysial axon terminals.

Animals↗

PubMed1994

Dynorphin-A and vasopressin release in the rat: a structure-activity study.

The effects on vasopressin (VP) release of three dynorphin-A fragments and two antidynorphin antisera were tested in vivo and in vitro. In vivo, the order of potency to inhibit VP release 30 min upon i.c.v. injection was: dynorphin-A-(1-17) > dynorphin-A-(1-13) > dynorphin-A-(1-8). l.c.v. co-administration of 10 nmoles of the specific endopeptidase-inhibitor cFPAAF-pAB and dynorphin-A-(1-8) also suppressed VP secretion. Dynorphin-A-(1-17) antiserum enhanced VP release 20 and 60 min after i.c.v. injection. The antiserum that recognized dynorphin-A-(1-13) elevated VP plasma levels at 60 min post-injection. In vitro, dynorphin-A-(1-8) suppressed electrically evoked VP release from the isolated neural lobe. VP release was not affected by dynorphin-A-(1-13), dynorphin-A-(1-17), naloxone, or by the anti-dynorphin antisera. These data indicate that dynorphin-A-(1-17), rather than dynorphin-A-(1-8), plays a role in the centrally located control of neurohypophysial VP release, whereas dynorphin-A-(1-8) is involved in the control located in the posterior pituitary. The synthetic intermediate fragment dynorphin-A-(1-13) appears to affect VP release both centrally and peripherally.

Analysis of Variance↗

PubMed1991

The opioid receptor subtypes mu and kappa, but not delta, are involved in the control of the vasopressin and oxytocin release in the rat.

The effects of highly selective agonists and antagonists to the mu-, delta- and kappa-opioid receptor subtypes were studied on the vasopressin and oxytocin release in 24 h water-deprived male rats. The delta-agonist [D-Pen2,D-Pen5]enkephalin (dose range 0.01-5 mg/kg) did not affect plasma levels of either hormone 30 min after s.c. administration, whereas the mu-agonist DALDA (H-Tyr-D-Arg-Phe-Lys-NH2) over the same dose range strongly inhibited the release of both vasopressin and oxytocin, an effect that was maximal 30-60 min after s.c. injection. The same effect was found for s.c. administration of the kappa-agonist U-69,593. Intracerebroventricular (i.c.v.) administration of DALDA (0.5 and 5 micrograms/kg) but not U-69,593 suppressed both plasma hormone levels 30 min after injection. Also the effects of selective antagonists were tested over the s.c. dose range of 0.01-1 mg/kg. Whereas both the kappa-selective antagonist nor-binaltorphimine and the relatively mu-selective antagonist naloxone elevated oxytocin plasma levels (peak at 15 and 30 min after injection, respectively), the delta-selective antagonist naltrindole was without any effect. Nor-binaltorphimine, naloxone, and naltrindole did not affect vasopressin release. When the antagonists were administered i.c.v. (dose range 2.5-25 micrograms/kg), only the kappa-antagonist nor-binaltorphimine enhanced oxytocin and vasopressin release 30 min after injection. In conclusion, both mu- and kappa-opioid receptors are involved in the regulation of the secretion of vasopressin and oxytocin from the rat neural lobe; in contrast, delta-opioid receptors do not play a role.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1991

Pharmacological assessment of the site of action of opioids on the release of vasopressin and oxytocin in the rat.

Naloxone and its congener, methyl naloxone, were given subcutaneously (s.c.) or centrally (i.c.v.) to 24-h water-deprived male rats 30 min prior to decapitation and the effect on plasma levels of vasopressin (VP) and oxytocin (OT) was studied. The potency of s.c. applied methyl naloxone to increase plasma OT levels did not differ from that of naloxone. Injected i.c.v., neither methyl naloxone nor naloxone had a clear effect and they antagonized i.c.v. co-administered dynorphin A-(1-13) equipotently. Methyl naloxone or naloxone, s.c., antagonized the inhibitory action of simultaneous dynorphin A-(1-13) and beta-endorphin-(1-31) given i.c.v., although higher doses of methyl naloxone were required. The data indicate that the sites of inhibition of neurohypophysial hormone release due to beta-endorphin-(1-31) are more likely to be located mostly within the blood-brain barrier, to which methyl naloxone has less ready access, than are the sites of inhibition due to dynorphin A-(1-13).

Animals↗

PubMed1991

Solid-phase extraction of plasma vasopressin: evaluation, validation and application.

A new solid-phase extraction method using octyl-silica columns to extract vasopressin-like immunoreactivity from plasma has been developed. The extraction was followed by a radioimmunoassay on the vacuum-dried extracts, which were reconstituted in assay buffer. The total recovery of synthetic vasopressin was ca. 100%. Based on co-elution with synthetic vasopressin after separation by reversed-phase high-performance liquid chromatography of plasma extracts from normal Wistar and Brattleboro rats, and the cross-reactivity of the antiserum used in the radioimmunoassay system, the extracted material was found to be indistinguishable from authentic vasopressin. Unknown experimental samples were interpolated on a standard curve established in "zero" plasma (plasma derived from rats subjected to waterload) spiked with known amounts of synthetic vasopressin, and not on a standard curve established in assay buffer. The limit of detection was 1 fmol of vasopressin equivalent per millilitre. The intra- and inter-assay coefficients of variance were 10-16% and 16%, respectively. The procedure reliably showed that osmotic challenge and 24-h dehydration increased, whereas ethanol ingestion decreased vasopressin-like immunoreactivity plasma levels in the rat, compared with normally hydrated controls.

Animals↗

PubMedDate not supplied

Effect of bile salts on in vitro gallbladder motility: preliminary study.

Impaired postprandial gallbladder emptying may be an important factor in cholesterol crystals precipitation and subsequent gallstone formation. We previously found strongly increased bile salt concentrations in gallbladder bile of gallstone patients with weak (< 50% fasting volume) postprandial gallbladder contraction compared to patients with strong (> 50%) postprandial contraction. Therefore, we studied potential effects of various conjugated and unconjugated bile salts with different relative hydrophobicity on in vitro contractility of gallbladder muscle strips obtained at cholecystectomy. Strips were incubated 5 min with bile salt at concentrations of 10(-8)-10(-4)M. The effect of 10(-3)M acetylcholine was measured and related to preincubation control value. Bile salts used were, in order of increasing hydrophobicity: tauroursodeoxy-, ursodeoxy-, tauro-, taurodeoxy- and deoxycholate. Ursodeoxy- and tauroursodeoxycholate did not significantly reduce gallbladder contractility. Taurocholate significantly reduced contractility at concentrations of 10(-6) M and higher, taurodeoxycholate at 10(-7) M and higher and deoxycholate at 10(-5) M and higher. Contractility induced by acetylcholine 10(-3) M at a bile salt concentration of 10(-4) M was 66.0 +/- 11.7% (taurocholate), 50.2 +/- 6.2% (deoxycholate) and 44.8 +/- 11.5% (taurodeoxycholate) of control. The effect of bile salts correlated with their relative hydrophobicity (r = -0.97; p < 0.01). Suppressing effects on gallbladder muscle strip contractility were long lasting and remained after rinsing. Results show that bile salts in the physiological dose range inhibit in vitro gallbladder contraction. If this mechanism exists in vivo, it may have important implications for gallbladder motility regulation.

Acetylcholine↗