Elastase,collagenase and the radial elastic properties of arteries

Summary Studies were performed on 203 pairs of dog carotid arteries subjected to unidirectional radial compression. Treatment with 80 U/ml purified elastase for 90 min decreased radial stress, but treatment with 640 U/ml collagenase for 90 min did not. These data suggest that elastin, but not collagen, contributes to wall resistance to radial compression.     

Synergistic effect of acute hypoxia on flow-induced release of ATP from cultured endothelial cells

Human umbilical vein endothelial cells (HUVECs) in primary cultures were perfused under normoxic or hypoxic conditions. These cells were stimulated twice for 3 min by increased flow (from 0.5 to 3.0 ml/min). Under hypoxic conditions the basal release of ATP was the same as under normoxic conditions, but during increased flow the release was greater (0.58±0.07>0.32±0.04 pmoles/ml/10⁶ cells (+78%), for the first period of stimulation; 0.39±0.05>0.22±0.03 pmoles/ml/10⁶ cells (+79%) for the second period). Further experiments with sequential increments in flow rate showed that under both normoxic and hypoxic conditions, a positive correlation existed between ATP release and the rate of flow but there was always more ATP released under hypoxic conditions regardless of the flow rate.HUVECs in secondary culture (second passage) were similarly stimulated. No differences were observed between normoxic and hypoxic conditions. In both cases, the quantity of ATP released during high flow (0.050±0.004 pmoles/ml/10⁶ cells) was significantly smaller than the quantity of ATP released during low flow (0.09±0.01 pmoles/ml/10⁶ cells).To conclude, since hypoxia alone did not affect ATP release, there appears to be a synergistic relationship between increased shear stress and hypoxia in the stimulation of ATP release from HUVECs. Moreover, the release of ATP under these conditions seems to be a property of highly differentiated endothelial cells.     

Production of butyrylcholinesterase by Caco-2 cells: lack of relationship with triglyceride production

Elevated levels of butyrylcholinesterase activity occur under a number of hypertriglyceridemic conditions, including diabetes and obesity. This study examines whether butyrylcholinesterase activity has a direct effect on triglyceride production, using Caco-2 cells, a human intestinal adenocarcinoma cell line. Caco-2 cells were incubated with 500 μM oleate to stimulate triglyceride production, and butyrylcholinesterase activity was measured in the cellular homogenate. Butyrylcholinesterase activity was approximately 3 × 10^-3 mmol/min per milligram protein. Although triglyceride production increased by almost five-fold after 18 h of stimulation with oleate, butyrylcholinesterase activity was not increased. Furthermore, inhibition of butyrylcholinesterase activity using 1 mM tetraisopropylpyrophosphoramide did not significantly affect triglyceride production or secretion. Human insulin (100 μU/ml) increased the production of butyrylcholinesterase without increasing triglyceride production. This demonstrates that stimulation of fatty acid production and butyrylcholinesterase activity occur by independent mechanisms and suggests that their correlation in hyperlipidemic conditions is not due to a direct relationship in production in situ. Received 23 April 2001; received after revision 25 May 2001; accepted 20 June 2001     

Adrenocortical metabolism and plasma corticosterone in soman intoxicated rabbits

The organophosphate neurotoxin soman produced impairments in adrenocortical RNA and protein metabolism. Fasciculate and reticular cell RNA and protein contents were suppressed with sublethal to acutely lethal dosages (20, 30 and 40 micrograms/kg, s.c.) during the acute excitatory phase of intoxication and at 6-8 h post injection. All three dosages produced ca 90% inactivation of plasma cholinesterase. A transient elevation of plasma corticosterone occurred with 20 micrograms/kg soman whereas there was a protracted increase with 30 micrograms/kg. Corticosterone was not significantly elevated with 40 micrograms/kg, but death occurred at 13 +/- 4 min. Thus, the magnitude and/or nature of soman-induced metabolic impairments does not appear to prevent adrenal activation.     

β2-integrins mediate a novel form of chemoresistance in cycloheximide-induced U937 apoptosis

In this study with cycloheximide (CHX, an inhibitor of protein synthesis) and the human leukaemic cell line U937, a novel form of chemoresistance, which we termed sudden drug resistance (SDR), was identified using Hoechst33258 staining, Western blott and DNA Ladder. CHX^high (10–100 g/ml)-induced apoptosis can spontaneously subside after 4–6 h or can be inhibited by short-term preincubation with CHXlow (2.5 g/ml). Unlike typical multidrug resistance, SDR is not caused by reduced drug accumulation or altered protein expression, and may be associated with a non-P-glycoprotein mechanism. To uncover this underlying mechanism, we focused on U937 cell aggregation promoted by CHX, because cell adhesion has been suggested to influence cell survival and prevent apoptosis. EDTA, or anti-CD18 monoclonal antibody, but not EGTA, acetylsalicylic acid or RGDS tetrapeptide, abrogated this homotypic aggregation and greatly increased CHX-induced apoptosis in a time-dependent manner, while fibrinogen and soluble intercellular adhesion molecule-1 exerted opposite effects. These results establish that ₂-integrin engagement is a key mediator of SDR, although it may be non-exclusive. This finding supplements the classical basis of chemoresistance and may provide another opportunity for improved leukemia therapy.Received 15 April 2004; received after revision 18 May 2004; accepted 21 June 2004     

Stimulation of histamine synthesis in rat mast cells by compound 48/80 in vitro

The histamine content of rat peritoneal fluid cells is doubled within 20 min by 0.5 microgram/ml of compound 48/80. Histamine catabolism inhibitors do not reproduce this effect; cells pre-incubated with alpha-fluoromethylhistidine are unresponsive to compound 48/80 which therefore activates pre-formed histidine decarboxylase rather than 'inducing' it. Non-mast cells showed no change after treatment with compound 48/80.     

Capillary blood flow in the testes and testosterone secretion in the starved rat

Summary The average capillary blood flow in the testes was found to be 181 l/min/g testis tissue (n=19) in rats starved for 5 days and 273 l/min/g (n=18, p<0.01) in the control group. Plasma testosterone was significantly decreased in the starved animals (1.00±0.06 ng/ml vs 5.43±0.63 ng/ml). When starved and control rats were stimulated with human chorion gonadotropin, testosterone values in plasma were greatly increased in both groups. The capillary blood flow was not altered. The data indicate that human chorion gonadotropin can stimulate testosterone production in the starved rat without influencing the reduced capillary blood flow.     

Oxygen consumption in human placental lobules in vitro (survival maintained by perfusion)]

The average oxygen utilization coefficient of 26% is calculated from PO2 determinations during the perfusion of the fetal circulation of isolated human term placental lobules. The oxygen consumption approximates 0.4 ml/min/kg placenta when Earle's salt solution is used as a perfusion medium and is equilibrated with a gas mixture containing 5% O2. The oxygen consumption reaches 2 ml/min/kg when the same Earle's solution is equilibrated with a gas mixture containing 95% O2.     

Infusion of a novel peptide, PHI, in man. Pharmacokinetics and effect on gastric secretion

PHI, infused in man, achieved plateau plasma levels of 297 pmoles/1. The plasma half life was 3.1 min, metabolic clearance rate was 16.4 ml/kg/min and estimated volume of distribution was 73.2 ml/kg. No subjective side effects were noted during the infusion and there was no significant alteration in submaximal pentagastrin stimulated gastric acid or pepsin secretion.     

Oxytocin antagonism of hypothalamic-induced angina-like ECG changes and pressor effects in the cat

Electrical stimulation of a specific site in the lateral hypothalamus of the cat, in a region posterior to Hess' defense area, results in pressor effects and angina-like ECG changes which consist either of T-wave inversion and ST-segment prolongation or in the appearance of tall T-waves. Oxytocin (10 U, i.v.) administered 15 min prior to stimulation, prevents the former ECG changes and BP rise in 90%, and the latter ECG changes and BP rise in 50% of the animals.     

Inhibition by cortisol of the favourable action of lysine-vasopressin on the growth of hela cell cultures

Résumé L'action stimulante de 25 µU/ml de lysinevasopressine de synthèse sur la croissance des cultures de cellules HeLa est inhibée par l'addition de 1 µg/ml de cortisol.     

Comparison of different techniques for semiquantitative detection of HBV DNA by PCR

Conclusions Our results indicate that the choice of the probe for ELOSA is of major concern. In our panel we had seven sera which contained about 100 molecules/ml and further five sera which contained less than 1000 molecules/ml. Most of them were detected by the long probe but not by the short probe. When PCR for the S-or PreS-gene was included it was possible to detec all 24 HBV-positive sera (not shown) by ELOSA. The reliable lower quantification limit for the long probe is 250 molecules/ml and for the short probe 2500 molecules/ml. Surprisingly, chemiluminescence did not produce better qualitative or quantitative results. The data suggest that the usage of several replicates allows relative quantification in most cases. One possible drawback we see is the hybridization efficiency. Six of our positive samples showed great differences between the number of target molecules suggested by agarose gel electrophoreses or by hybridization (Southern blot or ELOSA). All of them contained more than 10⁶ molecules/ml. For these cases and for the samples where the short probe and the long probe gave discordant result (2 cases) we think that competitive PCR will be the method of choice, but in most cases ELOSA with the long probe gives reliable results and is highly sensitive.     

Effects of ethanol and acetaldehyde on cultured pre-implantation mouse embryos

Pre-implantation 2-cell stage mouse embryos, obtained from superovulated CF-1 mice, were exposed to ethanol and acetaldehyde through the culture medium for 60 min followed by a 105-h incubation period. Control and ethanol exposed embryos survived equally well in ethanol concentrations as high as 800 mg/100 ml medium and acetaldehyde levels up to 10 mg/100 ml medium.     

A contribution to the evaluation of the glucose tolerance test

Zusammenfassung Es wird eine abgeänderte Formel für die Berechnung der Glukose-Clearance nach intravenöser Applikation vorgeschlagen.Das arithmetische Mittel und die Standardabweichung der Clearancewerte bei Gesunden betragen 220,56±54,9 ml/min, das heisst 3,29±0,92 ml/min/kg Körpergewicht.     

Effects of ethanol and acetaldehyde on cultured pre-implantation mouse embryos

Summary Pre-implantation 2-cell stage mouse embryos, obtained from superovulated CF-1 mice, were exposed to ethanol and acetaldehyde through the culture medium for 60 min followed by a 105-h incubation period. Control and ethanol exposed embryos survived equally well in ethanol concentrations as high as 800mg/100 ml medium and acetaldehyde levels up to 10 mg/100 ml medium.     

Regulation of ammonia uptake in aspergillus nidulans.

The ammonia uptake in A. nidulans was found to be linear for about 20 min, and was proportional up to 1.5 mg/ml dry cell density. The transport of ammonia does not involve energy. Normal and biotin deficient A. nidulans showed an identical Km-values of 10.26 X 10(-5) M ammonia for uptake. The uptake of ammonium ion has been shown to be regulated by the intracellular concentration of ammonia.     

Modification of the action of pentagastrin on acid secretion by botulinum toxin.

I.v botulinum toxin after 60-90 min abolished the dose-response relationship between pentagastrin and gastric acid secretion in anesthetized rats and guinea-pigs. The toxin reduced but did not abolish the acid stimulatory effect of histamine. As expected, the acid response to vagal stimulation was abolished and that to methacholine in rats was unaltered by the toxin.     

Periodate oxidation of lymphocyte membrane glycoconjugates]

Mouse spleen cells free of erythrocytes were suspended in PBS at a concentration of 2 X 10(7) cells/ml and mixed with an equal volume of sodium periodate in PBS for 10 min. at 4 degrees C to give a final concentration of periodate ranging from 10(-4) M to 5 X 10(-3) M. The cells were then washed and suspended (60 X 10(6) ml) in PBS containing 3H-labelled sodium borohydrate and incubated for 30 min, at 23 degrees C. Following this, the cells were washed and the pellets treated with H2SO4 0.1 N for 60 min. at 80 degrees C. Compounds liberated by such treatment, were identified by chromatography as derivates of sialic acid. The data provide direct evidence that the mitogenic effect of sodium periodate is associated to the oxidation of the sialic acid residues on the lymphocyte membrane.     

The effect of actinomycin D on the production of viruses by the protoplasts of Chinese cabbage infected in vitro by the yellow mosaic virus of turnips]

Chinese Cabbage (Brassica sinensis L. var. Cantonner) protoplasts were infected by Turnip yellow mosaic virus (TYMV) and inoculated in the presence or absence of actinomycin D. Virus production was determined 40 hrs. after inoculation, the time required for the virus replication cycle to be terminated. While actinomycin D had no effect on TYMV production when present at a concentration of 1 microng/ml, a 50 to 80% inhibition of virus production was noticed at concentrations of the order of 5 to 10 microng/ml, and the inhibition reached 90% with 25 microng/ml.     

Role of a free-living amoeba from water, Acanthamoaba castellani, in the transport of naked or enveloped animal viruses

The trophozo?ts of Acanthamoeba castellanii are unable to adsorb poliovirus or vesicular stomatitis Virus. After encystment in medium containing respectively 5.4 x 10(8) and 3 x 10(9) P.F.U./ml cysts did not contain Viruses. These data do not agree with a current hypothesis by which water's free Amoeba could carry animal Viruses.