Catecholamine secretion from adrenal chromaffin cells of the toad (Caudiverbera caudiverbera): effect of monensin.

1. Stimulus-secretion coupling studies were carried out on adrenal chromaffin tissue from the toad. Catecholamine (CA) secretion was generated in response to acetylcholine (ACh) or high K+. 2. The response to ACh was found to be dependent on the presence of external Ca2+. The secretion induced by ACh or high K+ was inhibited by the Ca-channel blockers CoCl2 and nifedipine. 3. The specific Na+/H+ ionophore, monensin, induced a strong secretory response only if Na+ was present in the Ringer. Monensin's effect did not depend on external Ca2+ and was unaffected by the channel blockers tetrodotoxin or CoCl2. 4. Secretion induced by monensin was exocytotic as was shown by measuring ATP release using a photoluminescence, luciferine/luciferase assay. 5. In conclusion, in the toad, as in higher species, stimulus-secretion coupling involves Ca2+ entry from the external medium, possibly through voltage-dependent channels. Monensin is a potent secretagogue and the mechanism by which the ensuing elevation of intracellular Na+ concentration might induce a secretory response remains to be determined.


The effects of gamma-aminobutyric acid on voltage-clamped motoneurons of the lobster cardiac ganglion.

1. We examined the electrophysiological and pharmacological effects of GABA on voltage-clamped motoneurons of the lobster cardiac ganglion. 2. GABA caused a dose-dependent current (EC50 = 0.7 mM), which reversed at the estimated Cl- equilibrium potential. 3. The conductance activated by GABA was voltage-dependent, increasing as a non-linear function of depolarization. 4. A Na(+)-dependent GABA uptake mechanism was only weakly sensitive to nipecotic acid. 5. Picrotoxin inhibited the GABA response, but bicuculline had no effect. 6. We conclude that the effect of GABA in the lobster cardiac ganglion is similar to its effect on other crustacean neuromuscular tissues and on vertebrate GABAA receptors. 7. There appear to be differences among species with respect to the physiology and pharmacology of the Na(+)-dependent GABA transporter. 8. The effect of GABA is also similar to the ionic mechanism underlying the action of histamine in the cardiac ganglion.


The in vitro efficacy of reuterin on the culture and bloodstream forms of Trypanosoma brucei brucei.

1. The in vitro effects of a new antibiotic, reuterin, were determined for culture and bloodstream forms of Trypanosoma brucei brucei. It inhibited growth of the culture forms and motility, viability and DNA and protein syntheses of culture and bloodstream forms. 2. Reuterin administered with inhibitors of ribonucleotide reductase (hydroxyurea and deoxyadenosine) was synergistic only for growth of the culture form of the parasite. 3. Reuterin was trypanocidal at lower doses than DFMO, Mel B, and Suramin and was antagonistic when used with these drugs.


Aflatoxin and glutathione in domestic fowl (Gallus domesticus)--I. Glutathione elevation and attenuation by high dietary methionine.

1. Changes in hepatic and renal glutathione (GSH) and plasma aspartate aminotransferase (AST) following single or daily oral doses of aflatoxin B1 (AFB1, 2 mg/kg BW) or corn oil vehicle (1 ml/kg BW) were determined in male chickens (14-21-day-old). 2. Plasma AST and hepatic GSH increased 2 and 8 hr, respectively, following a single AFB1 dose. 3. Hepatic GSH continued to increase through 5 daily doses of AFB1, but there were no differences in AST levels on days 1-5. Feeding a diet containing 150% of NRC requirement for methionine attenuated the AFB1-induced increase in hepatic GSH. Renal GSH was unaffected by AFB1 or dietary treatment.


Mytilus-inhibitory peptide analogues isolated from the ganglia of a pulmonate mollusc, Achatina fulica.

1. Ten peptides that showed an inhibitory effect on phasic contraction of the ABRM of Mytilus were isolated from the ganglia of the African giant snail, Achatina fulica. 2. Seven of the peptides were shown to be hexapeptides having -Pro-Xaa-Phe-Val-NH2 as a common structure, which was previously shown to be a characteristic of Mytilus inhibitory peptides (MIPs). 3. The remaining three were pentadecapeptides, each of which consisted of two MIP-related hexapeptides linked by -Gly-Arg-Arg-.


Metal redistribution in largemouth bass (Micropterus salmoides) in response to restrainment stress and dietary cadmium: role of metallothionein and other metal-binding proteins.

1. Fish stressed by restrainment displayed elevated serum cortisol, copper and zinc levels; dietary cadmium had no effect. 2. Stress/dietary cadmium increased liver copper levels in a metal pool containing metallothionein and non-metallothionein proteins but decreased intestinal zinc bound as low molecular weight forms. 3. After restrainment, zinc losses occurred in dorsal skeletal muscle, ovary and spleen: copper decreased in intestine and pyloric caecum. 4. Dietary cadmium altered intestinal zinc distribution and raised hepatic Cu-binding protein levels but did not alter plasma zinc, copper or cortisol levels. 5. Alterations in zinc and copper concentrations during stress contrast with mammalian models.


Blood morphology in quails after poisoning with fenitrothion.

1. The purpose of this study was to determine changes in the blood of Pharaoh quails after fenitrothion intoxication, to define the duration of disturbances caused by intoxication and to examine possible sex differences in the birds' reaction to intoxication. 2. Fenitrothion reduced the number of erythrocytes, haemoglobin level and haematocrit value, but increased erythroblast and reticulocyte numbers. 3. Fenitrothion caused neutrophilic leucocytosis with lymphopaenia and eosinopaenia. 4. In males, changes in the blood appeared far earlier than in females and they underwent compensation earlier, i.e. 3 weeks after intoxication the majority of the haematological parameters reached values similar to those of the controls.


Neuropeptide F: primary structure from the tubellarian, Artioposthia triangulata.

1. A neuropeptide exhibiting vertebrate pancreatic polypeptide immunoreactivity has been isolated and sequenced from extracts of the terrestrial turbellarian, Artioposthia triangulata. 2. This neuropeptide, designated neuropeptide F, consists of 36 amino acid residues terminating in a phenylalaninamide. 3. The full primary structure was established as: KVVHLRPRSSFSSEDEYQIYLRNVSKYIQLYGRPRF.NH2. The molecular mass, deduced from this sequence, was 4433 Da. 4. This neuropeptide exhibits C-terminal homology with neuropeptide F (Moniezia expansa) and with the vertebrate neuropeptide Y/pancreatic polypeptide superfamily of which it may represent a phylogenetic precursor.


Acute toxicity of ammonia to Atlantic salmon (Salmo salar) parr.

1. Atlantic salmon (Salmo salar) parr were exposed to (NH4)2SO4 solutions in static systems with aerated, soft water for 96 hr. The 96 hr-LC50 for un-ionized ammonia (expressed as mg/l NH3-N) ranged from 0.031 (2.1 degrees C) to 0.111 (17.1 degrees C) at pH 6.0 and from 0.030 (1.8 degrees C) to 0.146 (12.5 degrees C) at pH 6.4. 2. No mortality, was found in a KCl solution and a physiological salt solution with chloride concentrations approximately equivalent to the chloride concentration in a NH4Cl solution giving 45% mortality, and to the ammonia concentration in a (NH4)2SO4 solution giving 35% mortality, all solutions tested at pH 6.0 and 2 degrees C.


Involvement of non-classical 5-HT receptor in serotonin and cisapride induced secretion in hen colon.

1. In hen colon 5-HT induces a tetrodotoxin-resistant, bumetanide-sensitive, chloride secretion, positively coupled with adenylate cyclase activity. 2. The 5-HT receptor mediating this response seems non-classical since it cannot be blocked by 5-HT1-like, 5-HT2 or 5-HT3 antagonists. 3. Effects are presented of new putative 5-HT agonists and antagonists on short circuit current and cord conductance in the hen colon, using the Ussing chamber technique. 4. The substituted benzamides, cisapride and BRL 24924, induced a dose-dependent short circuit currents but both with less potency than 5-HT. 5. Cisapride mediated this dose-dependent bumetanide sensitive response mainly by release of acetylcholine, since atropine reduced cisapride response by 70%. 6. Neither BRL 24924, 5-HTP-DP, ketanserin, ICS 205-930, prazosin, yohimbine, atropine nor piroxicam, covering the 5-HT1P, 5-HT2P, 5-HT2, 5-HT3, 5-HT4, adrenergic and muscarinic receptor types and the prostaglandin synthesis, altered 5-HT induced increases in short circuit current and cord conductance. 7. Results suggest (a) cisapride mediates it's response mainly by releasing acetylcholine, which then stimulates muscarinic receptors to release 5-HT. (b) Involvement of a non-classical 5-HT receptor subtype in 5-HT induced chloride secretion in hen colon.


The effects of ethanol on intracellular potassium and the membrane potential of an identified crab motor axon.

1. Observations were made on the fast bender excitor axon in autotomized crab walking limbs bathed in normal crab saline, and in salines made up with 0.2 M sucrose or 0.2 M ethanol. Microelectrode techniques were used to measure the resting membrane potential and the intracellular level of potassium. 2. Sucrose-saline had little effect on the membrane potential or the intracellular level of potassium. Ethanol-saline hyperpolarized the membrane potential by about 3 mV and increased the level of intracellular potassium. 3. The ethanol-induced changes in intracellular potassium levels and membrane potential took place with the same time course. Further, the changes in membrane potential could be accounted for by changes in the equilibrium potential for potassium, Ek as predicted by the Nernst equation.


Furosemide decreases eggshell thickness and inhibits 45Ca2+ uptake by a subcellular fraction of eggshell gland mucosa of the domestic fowl.

1. Administration of furosemide to egg-laying domestic fowl (single p.o. dose 100 mg/bird) caused a decrease in the thickness of the shell of eggs laid the next day. 2. Furosemide administration in vivo caused a 37% decrease in the uptake of 45Ca2+ by a subcellular fraction of the eggshell gland mucosa (mainly composed of cell fragments and plasma membranes). 3. Furosemide treatment did not affect calcium concentrations in plasma or shell gland fluid but did cause a significant increase in the calcium concentration in shell gland mucosa. 4. It is concluded that the eggshell-thinning effect of furosemide is localized to an inhibitory effect on plasma membrane calcium transport in the eggshell gland mucosa. 5. These findings are discussed and compared with the effects of other drugs and toxic substances known to influence eggshell formation in birds.


Receptors mediating the actions of 5-hydroxytryptamine on the isolated retractor muscle of the penis preparations from Archachatina marginata (Swainson).

1. The effects of some 5-hydroxytryptamine (5-HT) receptor agonist and antagonists on the isolated retractor muscle of the penis of Archachatina marginata were investigated. 2. Both 5-hydroxytryptamine and lysergic acid diethylamide (LSD) contracted the preparations. The mean pD2 values obtained for 5-HT and LSD were 5.26 +/- 0.21 (N = 10) and 6.3 +/- 0.36 (N = 8) respectively. 3. 5-Methoxy, N,N,-dimethyltryptamine (5-MeODMT; N = 6) trifluoromethylphenylpiperazine (TFMPP; N = 6) alpha-methyl 5-hydroxytryptamine (alpha-Me-5-HT); N = 6) and both L and DL-5-hydroxytryptophan (L-5-HTP; N = 6, DL-5-HTP; N = 6) did not contract the preparations even when concentrations up to 10(-4) M were administered. 4. Ketanserin (10(-8)-10(-6) M) produced rightward shift of the concentration-response curve of 5-HT. (pA2 = 8.0 +/- 0.25, slope = 0.34 +/- 0.03; N = 6). 5. The contractions of the RMP to 5-HT were not antagonised by either LY53857 or ICS 205-930. 6. The present study does not reveal the presence of 5-HT1-like or 5HT2 receptors on the RMP. However 5-HT3 receptors do not seem to exist on this invertebrate smooth muscle.


The most primitive heart in the animal kingdom contains the atrial natriuretic peptide hormonal system.

1. The newly described atrial natriuretic peptide hormonal system appears to play an important role in the endocrine control of sodium and water metabolism in human and vertebrate animals, but neither atrial natriuretic factor (ANF, C-terminal amino acids (a.a.) 99-126 a.a. prohormone) nor the rest of the ANF prohormone have ever been demonstrated in the heart of an invertebrate. 2. The present investigation was designed to determine whether the earthworm, Lumbricus terrestis, the first animal in the phylogenic tree with any form of heart, has either ANF and/or the 98 a.a. N-terminus of the ANF prohormone. 3. Both an ANF-like peptide (189 +/- 32 ng/g of tissue) and the N-terminus of the ANF prohormone-like peptide (1985 +/- 27 ng/g of tissue) were present in the earthworm heart at concentrations significantly higher (P less than 0.001) than in rat (Rattus norvegicus) heart ventricles. 4. This newly-described hormonal system, thus, appears to be present in a much larger proportion of the animal kingdom than previously thought, including invertebrates as well as vertebrates.


Acute toxicity of ozone against morphology of gill and erythrocytes of Japanese charr (Salvelinus leucomaenis).

1. Acute toxicity of ozone exposure to Japanese charr (Salvelinus leucomaenis) was studied histopathologically, and hematologically on gill tissue and red blood cells (RBC) under different ozone concentrations (0-0.7 ppm). 2. Exposure of ozone above 0.7 ppm led to characteristic symptoms and all died of choking in 30 min. 3. Many swollen RBC were seen under the scanning electron microscope. 4. RBC congestion was serious in the gill where degeneration of lamellar epithelium was observed. However, injury to chloride cells was not clear.


Binding of the benzodiazepine ligand [3H]-RO 15-1788 to membrane preparations of the rabbit and turtle retina.

1. We have studied the binding of [3H]-RO 15-1788 to membrane preparations of the retina of rabbit (Lepus cunicula) and turtle (Pseudemys scripta elegans). 2. In both species, [3H]-RO 15-1788 binding was maximal at 0 degrees C and decreased with increasing temperature. It was saturable, protein concentration-dependent and specific. Flunitrazepam, unlabelled RO 15-1788 and ethyl-beta-carboline were the most effective displacers, whereas RO 5,4864 was ineffective. 3. In both turtle and rabbit retina, Scatchard analysis indicated the presence of a single binding site for [3H]-RO 15-1788. The KD was 0.75 nM in both turtle and rabbit, while the Bmax were 520 and 250 fmol/mg protein in turtle and rabbit respectively. A study of the association rate of [3H]-RO 15-1788 binding revealed faster kinetics in turtle, as compared to rabbit.


Possible site of action of 2-methylserotonin in inducing relaxation of acetylcholine-induced contraction in the molluscan (Mytilus edulis) smooth muscle.

1. The present study investigated the presence of 5-HT3 receptor using 2-methylserotonin (2-Me-5-HT) in the smooth muscle of Mytilus ABRM. 2. 2-Me-5-HT relaxed the acetylcholine-induced contraction in a dose-dependent manner ranging from 10(-6) to 3 x 10(-4) M (pD2 = 5.55 +/- 0.32). 3. 2-Me-5-HT-induced relaxation was antagonized by 3 x 10(-5) M ketanserin in a competitive manner (pA2 = 5.14 +/- 0.1), but not by cypropheptadine, mianserin, MDL 72222 or ICS 205-930 at a concentration of 3 x 10(-5) M. 4. 2-Me-5-HT (3 x 10(-4) M) did not alter the content of cyclic AMP and cyclic GMP in the ABRM. 5. These findings suggested that the 2-Me-5-HT-induced relaxation was mediated through 5-HT2-like receptors and was not linked to cyclic AMP or GMP systems, and, further, that 5-HT3 receptor subtype was not present in the ABRM.


L-carnitine protects fish against acute ammonia toxicity.

1. Juvenile chinook salmon (Oncorhynchus tshawytscha) were injected intraperitoneally (i.p.) with 0.25 M mannitol followed 1 hr later by an i.p. challenge of ammonium acetate. 2. At 10.75 mmol ammonium acetate/kg body weight, 98% of the fish showed signs of ammonium toxicity and 69% died. 3. Substitution of L-carnitine (10-16 mmol/kg) for mannitol afforded striking protection from the subsequent challenge with ammonium acetate; 67% showed no signs of ammonia toxicity and only 4% died. 4. Of other quaternary amines tested, trimethylamine oxide also afforded protection, but betaine and choline did not.


Morphological changes on nerves and histopathological effects on liver and kidney of rats by pentachlorophenol (PCP).

1. The chronic toxicity of pentachlorophenol (PCP) was studied in rats after 90-120 days of oral administration ad libitum of 0.3-3 mM PCP aqueous solutions. 2. Morphological studies of their sciatic nerves were performed by optical and electron microscopy. 3. They showed degenerative changes in about 10% of the A and B type of nerve fibers. 4. The myelin sheath was discontinued by complete separation in several concentric rings while some other parts of the nerve exhibited a variable loss of neurotubules, neurofilaments and other axoplasmic components. 5. However, the C type of nerve fibers, the blood vessels and the perineurium did not show any morphologic alteration. 6. It was also found that in the liver PCP caused hemodynamic vein changes and injury in the hepatocytes such as cellular swelling and vacuolar degeneration. 7. The damage in the kidney occurred primarily in the glomeruli and secondarily in the proximal tubules causing turbid tumefaction and the formation of casts in the tubular lumen.


Inhibition of the sodium transport by pentachlorophenol (PCP) in toad skin (Pleurodema thaul).

1. The effects of pentachlorophenol (PCP) were studied in vitro on an active ionic transporting epithelium. Ussing's technique was applied on the isolated Pleurodema thaul skin. 2. Concentrations of PCP in the range 0.003-0.043 mM caused an irreversible dose-dependent inhibition in both the short-circuit current and the potential difference. 3. Parameters of an electrical equivalent circuit were calculated applying the Isaacon's amiloride test. 4. It was also shown that PCP produced a significant increase in the O2 consumption of the skin. 5. The inhibitory action of PCP on active sodium transport in terms of the equivalent electrical circuit and the increased oxygen consumption points to an uncoupling action of PCP on the oxidative phosphorylation.


Effects of chromium during pH change on blood coagulation in Tilapia sparrmanii (Cichlidae).

1. Tilapia sparrmanii was exposed to 0.098 mg/l potassium dichromate. 2. The effect of chromium on blood coagulation was measured with a thrombelastograph at a pH of 5, 7.4 and 9 over 96 hr, as well as an uncontrolled pH after 2 weeks of exposure. 3. The prolonged kinetic times and reduced maximal amplitudes at a pH of 7, 4 and 9, differed significantly from the control values. 4. Thrombelastograms reflected a decrease in the clotting ability of fish blood, with an increase in pH, which is typical of thrombocytopenia. 5. It was evident that the exposure of fish to chromium led to clotting defects that caused internal bleeding.


The effect of hexavalent chromium at different pH values on the haematology of Tilapia sparrmanii (Cichlidae).

1. The haematology of Tilapia sparrmanii (Smith) was investigated after exposure to 0.098 mg.l-1 hexavalent chromium at three different pH values. 2. Statistically significant changes occurred between the values of parameters of experimental and control fish. 3. At lower pH values erythrocytosis and leucocytosis were evident. 4. At an alkaline (9) pH anaemic and leucopenic conditions were observed. 5. T. sparrmanii adapted to chronic exposure to hexavalent chromium.


W-7, a calmodulin antagonist, and contracture of malignant hyperthermia susceptible skeletal muscle.

1. In malignant hyperthermia susceptible muscle fibers, the calmodulin antagonist, W-7 (10 microM), evoked contractures and potentiated halothene (3%) induced contracture. No effect was seen at 0.1 or 1.0 microM) W-7. 2. Dantrolene sodium (6 microM) prevented and reversed W-7 induced contracture: nifedipine did not. 3. In chemically skinned fibers, 10 microM, 1.0 microM, and 0.1 microM W-7 released 100%, 30%, and 10% of stored calcium respectively, and the effect of 10 microM W-7 was irreversible in that the SR was unable to re-sequestor calcium after exposure to the drug. 4. The release of calcium by W-7 was not prevented by exogenously added calmodulin (3 microM), nor mimicked by mastoparan (10 microM). 5. Calcium release by W-7 appears to be independent of calmodulin inhibition.


Beta 2-adrenergic regulation of urea permeability of the Bufo bufo bladder.

1. Isoprenaline strongly increases the urea permeability of the bladder of Bufo bufo. This effect is due to its interaction with beta 2-adrenoreceptors, activating, in turn, the adenyl cyclase. 2. In order to ensure the regulation of urea permeability, the isoprenaline effect is present even in pathophysiological conditions, inhibiting the vasopressin action.


The action of a series of glutamic acid analogues on Helix neuronal glutamate receptors.

1. Intracellular recordings were made from identified Helix central neurones, sensitive to L-glutamate. 2. Out of a range of substituted glutamate analogues, only the L- and D-isomers of thio-glutamate possessed clear glutamate-like activity. 3. On neurones excited by L-glutamate, the EC50 values for L-glutamate, gamma-thio-L-glutamate and gamma-thio-D-glutamate were 30 microM, 20 microM and greater than 1 mM, respectively. 4. On neurones inhibited by L-glutamate, the EC50 values for L-glutamate, gamma-thio-L-glutamate and gamma-thio-D-glutamate were 6.0 microM, 0.7 microM and greater than 200 microM, respectively. 5. It is concluded that, unlike the situation with thio derivatives of GABA, thio derivatives of glutamate possess potent glutamate-like activity.


Comparative assessment of the effect of aflatoxin B1 on hepatic dysfunction in some mammalian and avian species.

1. Aflatoxin B1 (1.5 mg/kg body weight, i.p.) was administered to rats, mice, quail and chickens to examine the comparative effect on hepatic microsomal drug-metabolizing enzymes, cytosolic glutathione S-transferase and serum enzymes. 2. Administration of aflatoxin B1 to rats resulted in a significant decrease in microsomal cytochrome P-450, NADPH-cytochrome c reductase, activities of aminopyrine N-demethylase, aniline hydroxylase, cytosolic glutathione S-transferase and liver glutathione content. However, no significant changes in these parameters were seen in mice. 3. Quail showed a significant decrease in the content of cytochrome P-450 and the activities of aminopyrine N-demethylase, aniline hydroxylase and cytosolic glutathione S-transferase. A similar treatment did not affect these biotransformation enzymes in chickens. 4. The activities of serum enzymes, sorbitol dehydrogenase, alanine aminotransferase and aspartate aminotransferase were increased significantly in rats and quail. Mice exhibited a significant increase in the activities of sorbitol dehydrogenase and aspartate aminotransferase, while chickens showed a significant increase only in alanine aminotransferase.


Effects of naloxone on membrane potential of identified neurons of Helix aspersa.

1. The action of naloxone, at doses that antagonize the inhibitory actions of opioid agonists in mammals, was tested in identified cells of Helix aspersa. 2. Naloxone was able to modify the membrane potential of some cells without prior exposure to opiate agonists. Some cells were depolarized and others were hyperpolarized. 3. Met-enkephalin was also effective in eliciting changes in membrane potential of some cells, and failed in others, some of which responded to naloxone, and was able to partially antagonize the effect of naloxone. 4. The electrophysiological responsiveness of this preparation to opioid agonists and antagonists makes this mollusc a suitable model for the study of opioid dependence and naloxone pharmacology.


Effect of Bothrops asper (Fer-De-Lance) snake venom on erythrocyte membrane. A comparative study.

1. The effect of Bothrops asper venom on erythrocytes of different species, sheep erythrocyte ghost vesicles and liposomes made of phospholipids from sheep erythrocytes was studied. 2. B. asper venom had a direct hemolytic effect on mouse erythrocytes, but no lytic activity on goat, rabbit, horse, toad and sheep erythrocytes. Human erythrocytes were lysed only in the presence of bovine serum albumin and Ca2+. 3. Although the phospholipase A2 activity in the venom might be involved in the lytic effect on mouse erythrocytes, there is also evidence of direct lytic factor(s) acting in the absence of calcium ions. 4. Sheep erythrocytes were modified in order to study the basis of their resistance. Enhancement of membrane fluidity and variation of pH were of no consequence for the resistance of sheep erythrocytes to the action of venom. 5. The reorganization of membranes that takes place during ghost or liposome preparation made them susceptible to B. asper venom-induced disruption. Thus the resistance of sheep erythrocytes seems related to the accessibility of the target.


Aflatoxin and glutathione in domestic fowl (Gallus domesticus)--II. Effects on hepatic blood flow.

1. The effect of aflatoxin on plasma aspartate aminotransferase (AST), protein, and hepatic glutathione (GSH) and hepatic blood flow (perfusion), were determined in 3-week-old male chickens. 2. Daily aflatoxin gavage (2 mg/kg body wt, in corn oil) for 5 and 10 days elevated plasma AST and hepatic GSH, and depressed plasma protein and hepatic perfusion. Also, renal GSH was elevated after 10 days of aflatoxin treatment. 3. Birds given aflatoxin for 10 days followed by a 10-day recovery period exhibited tissue GSH, plasma AST and protein levels that were not different from control, but hepatic perfusion remained depressed.


Identification of a novel 5-HTN (Nematoda) receptor from Ascaris suum muscle.

1. The abilities of various serotonergic drugs to bind with the 5-HT receptor of Ascaris suum muscle and to affect cyclic AMP levels in muscle tissue were examined. 2. Ligands which selectively interact with either the 5-HT1 or the 5-HT2 receptor in mammalian systems interact with the 5-HT receptor from A. suum muscle and increase cyclic AMP levels. 3. No binding of 5-HT3 ligands to 5-HT receptors from A. suum muscle was observed. 4. The 5-HT receptor of A. suum muscle should be called the 5-HTN (for Nematoda) receptor because its pharmacological and biochemical behaviors were different from those of mammalian 5-HT receptors.


Combined effect of fascioliasis and diethylnitrosamine carcinogenesis on the activity of the rat liver monooxygenase system.

1. The activity of the rat liver monooxygenase system after single and combined treatment with Fasciola hepatica and diethylnitrosamine (DENA) has been studied in a 27-week experiment. 2. The changes in lever drug metabolism were due mainly to F. hepatica with DENA having a modulating effect only. 3. The results are discussed with reference to earlier data of ours concerning the effects of stimulation or inhibition of DENA-induced liver carcinogenesis against a background of acute or chronic fascioliasis.


Quantitative characterization of beta-adrenergic receptor subtypes in porcine adipocytes.

1. Two populations of beta-adrenergic receptor (beta AR) subtypes and their proportions were characterized in adipocytes isolated from subcutaneous adipose tissue of castrated male crossbred pigs (60-85 kg). 2. Specific binding of the radioligand 125I-iodopindolol (IPIN) to crude plasma membranes (70-90% of total binding) reached equilibrium conditions in 30 min (38 degrees C), was tissue concentration-dependent, stereospecific and saturable (bmax = 168 +/- 5.8 fmol/mg protein). 3. Displacement curves by ICI 89,406 were best-fit by a two site model (P less than 0.01) that indicated the presence of two receptor populations and selectivity of IPIN for the beta 2AR subtype. 4. Forty-five percent of the receptors had a high affinity for ICI 89,406, Ki = 2.27 +/- 0.68 nM and were classified as beta 1AR.


Serotonin-immunoreactive neurons in the ventral nerve cord of the larva of the Eastern spruce budworm, Choristoneura fumiferana.

1. Using indirect immunofluorescent method, the distribution of serotonin immunoreactivity was examined in the ventral nerve cord of the larva of the Eastern spruce budworm, Choristoneura fumiferana. 2. There were two pairs of serially homologous serotonin immunoreactive neurons per ganglion. 3. The subesophageal ganglion which develops from the fusion of three neuromeres had accordingly, six pairs of immunoreactive neurons. 4. The neurons were positioned ventrolaterally at the posterior end of the ganglia and distributed in a bilaterally symmetrical fashion. 5. The axonal processes from serotonin-immunoreactive neurons projected to the contralateral side of the hemiganglion through a ventral commissure and formed an extensive network of fibers on the dorsal side of each ganglion.


Ethanol-inducible microsomal aniline hydroxylase activity and cytochrome P-450 isozymes in adult hen liver.

1. Cytochrome P-450 was induced in adult hen liver by administering 15% ethanol in drinking water and compared with other inducers such as phenobarbital and beta-naphthoflavone. 2. Aniline was the only substrate whose turnover was induced by ethanol treatment when measured in the presence of 100 microM alpha-naphthoflavone. 3. The inhibitor alpha-naphthoflavone differentiated aniline and p-nitrophenol hydroxylase activities, while p-hydroxyphenyl imidazole and SKF differentiated p-nitrophenol hydroxylase activity between ethanol- and beta-naphthoflavone-induced microsomes. 4. Ethanol treatment also slightly induced some P-450 isozymes related to phenobarbital and beta-naphthoflavone inducers.


Impact of cadmium on the mummichog Fundulus heteroclitus and the role of calcium in suppressing heavy metal toxicity.

1. Freshwater adapted mummichogs (Fundulus heteroclitus) were exposed to cadmium. In soft water (less than or equal to 5 mg/l CaCO3), the 4-day safe (TL100) and sublethal (TL50) tolerance limits (TLs) for cadmium were 0.14 microgram/l and 12.2 micrograms/l, respectively. 2. Survival declined with increasing cadmium concentration and the length of exposure. The toxicity of cadmium was reduced in water with high calcium concentration (200 mg/l CaCO3). Pre-exposure to calcium also prolonged the survival in cadmium-containing water. 3. The mummichog appears to be extremely well suited for monitoring environmental cadmium poisoning.


Physiological properties of liver, gonads and muscle during maturation of female Atlantic salmon, Salmo salar. Comparison between a control and xenobiotics containing fish feed.

1. Atlantic salmon, Salmo salar, in their 4th year were maintained in a sea-based farm in the Baltic Sea, salinity 0.2-0.4%. The fish were fed a control diet or a diet containing, among the lipid-soluble xenobiotics, 60 parts of dioxin per 10(12) parts of fish oil. 2. In the fish that attained sexual maturity the liver and gonadal wet weight increased, but decreased after stripping of the roe. Vitellogenesis was also apparent as an elevated level of plasma vitellogenin which was higher after than 2 months prior to removal of the roe. 3. In muscle protein content was highest prior to the removal of the roe. RNA content decreased with time. Following the taking of the roe glycogen content and acid proteinase activity were elevated. 4. Comparison between the feeding groups showed, in the fish fed the experimental diet, a higher gonadal wet weight and plasma vitellogenin content, and in muscle, after stripping of the roe, a lower glycogen content and an elevated level of acid proteinase activity.


Chelating agent reversal of cadmium effects on ionic transport in the isolated frog skin (Rana temporaria).

1. The application of 1 mM CdCl2 to the outside surface of frog skin causes a large increase in the potential difference (PD) across the skin and in the short-circuit current (SCC); the subsequent addition of selected dithiocarbamate chelating agents (which by themselves have no effect on PD or SCC) restored both electrical parameters to values close to initial levels. 2. The response observed on addition of the chelating agents indicates that the effect of CdCl2 is reversible and that the complexed ions do not possess the ability to initiate corresponding changes in the transepithelial ion transport processes in the frog skin.


Effects of pressure and gaseous anesthetics on the aggregation of human blood platelets and marine sponge cells: similarities in responses.

1. Aggregation of blood platelets and marine sponge cells was inhibited by nitrous oxide (N2O) and helium (He) at elevated pressure but potentiated by xenon (Xe) despite the fact that Xe and N2O are equipotent gaseous anesthetics. 2. The above aggregations are dependent on an extracellular source of calcium. 3. Platelet aggregation induced by phorbol myristate is independent of extracellular calcium and was inhibited by both N2O and Xe and by high pressures of He. 4. The results argue against a common mechanism for Xe and N2O and suggest that pressure may affect calcium interactions with binding site.


The effects of clonidine and three 2-imidazoline derivatives on the secretion of protein and some electrolytes by rat submandibular and parotid glands.

1. Three imidazoline analogues of clonidine were potent secretagogues for the parotid and submandibular glands at relatively high doses. 2. Salivation in response to clonidine was completely abolished by prazosin, phentolamine, phenoxybenzamine and dihydroergotamine. 3. The gamma-type of proteins was secreted in response to three of the analogues, whereas with p-aminoclonidine the alpha-type of proteins was secreted by the submandibular gland. 4. Albumin was specifically secreted by the submandibular gland in response to clonidine but not to isoproterenol or phenylephrine.


Phase-shifting of a neuronal circadian pacemaker in Bulla gouldiana by pentylenetetrazol.

1. The convulsant agent pentylenetetrazol generates compound action potential activity from the circadian pacemaker cells in the Bulla retina. 2. The phase response curve to 3 hr pulses of pentylenetetrazol consists of only phase delays which occur following pulses delivered in the early subjective night. 3. Phase shifts to pentylenetetrazol are independent of extracellular calcium since they persist in a low-calcium EGTA solution.


The autonomic innervation of the large intestine of the toad (Bufo marinus).

1. A study was made of the pelvic and the splanchnic nerve supplies to the toad large intestine. 2. Stimulation of pelvic nerve fibres in the 9th and 10th spinal nerves caused a series of contractions of the circular muscle, only the first of which was abolished by hyoscine. The entire response was blocked by d-tubocurarine. The response was not affected by capsaicin treatment. 3. Stimulation of the splanchnic nerves caused a rapid contraction followed by a prolonged relaxation. The relaxation was abolished by bretylium. The contraction was selectively antagonised by prolonged exposure to capsaicin. Splanchnic nerve stimulation also caused a slow, prolonged excitation that was abolished by bretylium. 4. Application of adrenaline caused relaxation of circularly cut strips of large intestinal wall, whereas substance P, acetylcholine, 5-hydroxytryptamine, somatostatin and galanin caused contraction. 5. The results suggest that stimulation of the pelvic nerves releases acetylcholine and a non-cholinergic co-transmitter from peripheral postganglionic neurons. Both the inhibitory response to splanchnic nerve stimulation and the subsequent slow excitation appear to be mediated by adrenergic nerves. The rapid capsaicin-sensitive excitation is likely to be due to release of substance P from antidromically activated afferent nerve fibres in the splanchnic outflow.


Induction of cadmium tolerance in two clones of Daphnia magna straus.

1. The abilities of two different genotypes of Daphnia magna to develop cadmium resistance through physiological adaptation, after pre-exposure to sublethal concentrations of Cd, Zn and Cd/Zn mixtures, was investigated. 2. The induced elevation in cadmium tolerance was shown to be associated with an increase in the body concentration of metallothionein-like proteins. 3. The highly significant difference in acute responses to cadmium between the two clones reduced after pre-exposure. 4. Differences in cadmium tolerance were shown to be associated with differences in cadmium uptake.


Effects of a benzodiazepine on the muscle membrane architecture of the rat diaphragm. A freeze-fracture study.

1. Midazolam increases, stimulation-independently, the amount of intermembraneous particles on the sarcolemma of the muscle fibres of the diaphragm. 2. Midazolam does not affect the amount of orthogonal arrays of particles on the sarcolemma. 3. Possible mechanisms for the action of midazolam are discussed.


Penetration, excretion and metabolism of 14C malathion in susceptible and resistant strains of Plutella xylostella.

1. The rate of penetration 14C malathion into the larvae of the diamondback moth, Plutella xylostella L., was found to be significantly different in an R-strain and S-strain. The rate of penetration was more rapid in the R-strain during the first hour after treatment. 2. The rate of metabolism in vivo and the rate of excretion were also higher in the R-strain compared with the S-strain. 3. The main metabolites produced in both strains were malathion dicarboxylic acid and desmethyl malathion. 4. The mechanism of insecticide resistance in Plutella xylostella L. is multifactorial, and involves a higher rate of penetration into the larvae of the R-strain, a higher activity of enzymes involved in its metabolism, and a higher level of excretion of the toxic compounds from the body of the R-strain compared with the S-strain.


Influence of the nutritional state of Triatoma infestans over the insecticidal activity of DDT.

1. The influence of nutritional state over topical insecticidal activity of DDT in nymph II of Triatoma infestans was analyzed. 2. DDT LD50 for starved nymph II was more than 150 micrograms/insect, while for nymph II fed to repletion on artificial feeder, was 0.79(0.50-3.13) micrograms/insect and a similar result was obtained for nymph II fed on pigeons (1.09(0.36-7.55) micrograms/insect). 3. A very slow penetration rate of 14C-DDT was obtained in non fed insects (4.4% penetrated 46 hr after treatment) in correspondence with 0% of mortality. 8.5% of 14C-DDT was not recovered from the cuticle 2 hr after topication of fed nymph in correspondence with a 100% of mortality at 24 hr.


The dynamics of MS-222 anaesthesia in a marine teleost (Pagrus auratus: Sparidae).

1. MS-222 anaesthesia of the marine teleost Pagrus auratus caused a dose-dependent elevation in haematocrit. 2. At 60 mg/l MS-222 the elevated haematocrit was fully accounted for by erythrocyte swelling, but at 100 mg/l red cell count also increased. 3. Uptake of anaesthetic in all tissues was rapid and the rate of onset of anaesthesia was dose-dependent. 4. Stage III level of anaesthesia (loss of equilibrium) was correlated with critical levels of MS-222 in the blood and brain. 5. The presence of acetylated derivatives of MS-222 in the blood demonstrated degradation of the anaesthetic, and high levels in the liver and kidney suggested that these organs were the sites of drug metabolism.


Isolation and characterization of a Cd-binding protein from Allolobophora caliginosa (Annelida, Oligochaeta): distinction from metallothioneins.

1. One Cd-binding peak was detected after gel filtration chromatography on Sephadex G75 in an extract from Allolobophora caliginosa contaminated with Cd. 2. Two subsequent cation-exchange chromatographies allowed the isolation of a Cd-binding protein which was called Cd-BP14. This protein is a monomer with a molecular weight of 14 kDa and has an isoelectric point of 6.5. 3. Amino acid analysis showed the presence of a high level of aromatic amino acids and a lack of cysteine. 4. On the basis of these results we conclude that Cd-BP14 is different from metallothioneins described in mammals or other invertebrates.


Reductive biotransformation of xenobiotics by the sheep ruminal content.

1. Sheep ruminal content was able to reduce nitro groups from nitrobenzene and azo groups from dimethylamino-azobenzene. 2. Results might be of interest in relation to ruminants exposed to environmental chemicals via oral route. 3. Biotransformation of xenobiotics in rumen might give to deleterious products appearing later in meat and/or milk or harming the ruminant itself.


Comparison of cadmium, mercury and calcium accumulations by isolated hepatocytes of the small skate (Raja erinacea) and rat.

1. Accumulation of calcium, cadmium and mercury by isolated hepatocytes of the small skate (Raja erinacea) and rat was examined at 14 and 37 degrees C, respectively. 2. Metal uptakes by both species were biphasic, with rat cells accumulating more metal than the skate cells. 3. Total accumulation after 30 min was in the order: mercury = cadmium much greater than calcium. 4. In both species calcium and cadmium accumulations were reduced at 4 degrees C, while mercury accumulation was not. 5. Cd accumulation was increased by Cu and Hg in both species. 6. Hg accumulation was inhibited by Cu in both species, and increased by Cd only in the rat cells.


Differential haemotoxic effect of PCB congeners in the common shrimp, Crangon crangon.

1. Specimens of the common shrimp, Crangon crangon, were exposed to sub-acute logarithmically separated concentrations (ranging from 0.05 microgram to 500 micrograms l-1) of two structurally dissimilar PCB congeners (PCB 15 and PCB 77) for five days in vitro. 2. Mortality, recoverable haemolymph volume, haemocyte count, plasma protein and haemolymph osmolarity were determined for test animals together with levels of haemolymph cell prophenoloxidase, an indicator of immune potential. 3. Sub-acute concentrations of PCB 15, but not PCB 77, produced significant decreases in haemocyte count and phenoloxidase activity with a marked increase in recoverable haemolymph volume. 4. No effect was observed on haemolymph osmolarity or plasma protein levels with either PCB 77 or PCB 15.


Bioconcentration of atrazine in the banded tilapia, Tilapia sparrmanii.

1. The bioconcentration of atrazine was determined in the liver, muscle, heart, gonads and brain of Tilapia sparrmanii exposed to high concentrations of atrazine. 2. The highest concentrations were recorded in the ovaries (50.6 micrograms/g) and in the liver (40.1 +/- 5.5 micrograms/g). This may be attributed to the higher lipid content of these organs, while the liver also accumulates atrazine as a result of its detoxification function. 3. The bioaccumulation factors for atrazine in the liver, muscle, heart, gonads and brain ranged from 0.9 to 20.0. Bioconcentration of atrazine in banded tilapia was found to be low, even after exposure to external atrazine concentration much higher than detected in natural surface water.


L-sorbose does not cause hemolysis in dog erythrocytes with inherited high Na, K-ATPase activity.

1. The hemolytic effect of L-sorbose on canine erythrocytes characterized by inherited high Na, K-ATPase activity and a high potassium concentration (HK RBCs) was compared with that on normal canine erythrocytes (LK RBCs). 2. Dogs having HK RBCs (HK dogs) revealed no clinical and hematological changes after administration of L-sorbose, whereas normal dogs (LK dogs) developed severe hemolytic anemia associated with hemoglobinuria and marked decreases of erythrocyte ATP concentrations. 3. In vitro, L-sorbose induced hemolysis in LK RBCs along with the depression of both ATP and lactate formation in these cells, but not in HK RBCs. The inhibition of glycolysis by L-sorbose in LK RBCs, however, was not observed when glucose-6-phosphate was used as a substrate instead of glucose. 4. These results suggest that the disparity of susceptibility to sorbose-induced hemolysis may be due to the difference in erythrocyte metabolism between HK and LK RBCs, especially the high activity of hexokinase in HK cells, which was 2-fold greater than that in LK RBCs.


The metabolism of noradrenaline in the sheep and the effect of dry matter intake upon the production of a metabolite, urinary vanillylmandelic acid.

1. The metabolism of noradrenaline was investigated in the sheep using 3H-noradrenaline injected subcutaneously or intravenously. 2. Analysis of urine collected over 24 hr showed that the bulk of 3H-noradrenaline was rapidly degraded and excreted via the urine. The principle metabolite was the conjugated amine as well as monohydroxy phenyl glycol and vanillylmandelic acid (VMA) previously reported for the rat. 3. Urinary VMA from lambs of two breeds noted for differences in fat (Southdown and Suffolk) increased curvilinearly with dry matter intake and there was a significant breed difference in regression with intake. 4. It is feasible that differences in noradrenaline production could contribute to differences in body composition between breeds.


Duck lymphocytes--V. Transformation responses to phorbol ester and calcium ionophore.

1. Lymphocytes purified from duck blood and spleen were cultured in the presence of phorbol 12-myristate 13-acetate (PMA) and the calcium ionophore A23187. Stimulation was assessed by the incorporation of [3H]thymidine after 3 days' culture. 2. PMA stimulated over a wide range of concentrations, with maximum stimulation at final concentrations of 5 x 10(-7)-5 x 10(-8) M/litre. A23187 was effective in the range 5 x 10(-6)-5 x 10(-7) M/litre and also, in some experiments using spleen lymphocytes, at 5 x 10(-11)-5 x 10(-12) M/litre. 3. Synergism was observed between PMA and A23187, the pattern depending on the concentrations of these reagents employed. Synergism was also observed between PMA and suboptimum concentrations of phytohaemagglutinin (PHA), wheat germ agglutinin (WGA), pokeweed mitogen (PWM) and Bandeiraea simplicifolia seed extract (BSS), but not with concanavalin A (Con A), lentil lectin (LL) or Helix pomatia lectin (HP). Similarly, synergism occurred between A23187 and WGA or PWM, but not with PHA, BSS, Con A, LL or HP. 4. Mitomycin C and cycloheximide inhibited the response of duck lymphocytes to PMA, A23187 and lectins. Cyclosporin A inhibited responses to lectins but not to PMA or A23187. Neither hydrocortisone nor indomethacin inhibited responses to lectins, PMA or A23187. 5. These results indicate that activation of duck lymphocytes occurs by virtue of similar intracellular messenger pathways to those operating in mammalian lymphocytes.


Systematic desensitization of oral hypersensitivity in a patient with a closed head injury.

A 36-year-old man who had sustained a closed head injury displayed extreme fear of being stimulated in the oral cavity, of being presented with foods, and of swallowing. The patient's fear increased his muscle tone and hypersensitivity in the facial and oral area, thereby preventing assessment of his dysphagia. We describe the use of systematic desensitization to alleviate the patient's fear thus allowing successful completion of a videofluoroscopic barium swallow examination.


[The bactericidal efficiency of ultrasonic in the root canal].

The objective of this paper was to determine the quantitative bactericidal efficiency of ultrasonic in the root canal. Four test organisms found frequently in the root canal were compared. The result showed ultrasonic can kill the test organisms effectively in the root canals. The best bactericidal efficiency occurred at 4 or 5 min. of ultrasonic though the more efficiency as the time longer. Bactericidal irrigation could increase the bactericidal efficiency of ultrasonic and its bactericidal action similar to irrigation bactericidal property.


[Immediate effect and family history of the type I, II schizophrenia].

34 cases of the type I Schizophrenia and 30 cases of the type II schizophrenia had been studied in immediate effect and family history. Symptomatic changes were tested by SANS and SAPS. The results showed that type I group was more cases of prominent effect and social function revival, less the time of prominent effect, less meam daily in hospital, less the time of happening psychosis disease and the positive family history than the type II group. (P less than 0.005, P less than 0.05). But no statistical significance in comparison of the all-dose of antipsychotic drugs between the two groups (P greater than 0.05).


Defective monocyte oxidative metabolism in a child with Smith-Lemli-Opitz syndrome.

We present a patient with Smith-Lemli-Opitz syndrome with immunodeficiency. The patient suffered numerous infectious episodes, atopic dermatitis and wheezing. Immunological investigations demonstrated severely reduced oxidative burst-responsiveness of the blood monocytes, whereas chemotaxis, phagocytosis and interleukin-1 production were normal. Tests of neutrophils and lymphocytes were normal excluding previously described immune deficiency disorders. The father proved to have diminished monocyte oxidative metabolism as well, whereas the mother had normal monocyte function. The genetic and immunological aspects are discussed in relation to the syndrome.


The value of laparoscopy in the management of the impalpable cryptorchid testis.

From January 1981 till October 1991, 47 diagnostic laparoscopies were performed in 50 impalpable testicles. In total, 28 intra-abdominal testes were found. In 14 cases no testes were found, but a deferential duct and vessels were seen. In 7 cases the diagnosis of testicular agenesia was made. One laparoscopy was a technical failure. We bring our results of this safe and reliable procedure, and discuss our management. A review of treatment options for intra-abdominal testes is given.


Central obesity and hyperinsulinaemia in women are associated with polymorphism in the 5' flanking region of the human insulin gene.

Obesity is a multifactorial disease with a marked genetic component. The situation is further complicated by the heterogeneity of obesity demonstrated by the topographical distribution of body fat, e.g. upper body (central) and lower body (gluteal) obesity. Furthermore, the distribution of fat shows a stronger heritable tendency compared with total body fat. Central obesity is characterized by hyperinsulinaemia and insulin resistance, a feature in common with non-insulin dependent diabetes mellitus, hypertension and atherosclerosis. In order to study the molecular genetics of central obesity we have examined 56 severely obese (mean body mass index 40), unrelated British Caucasoid young non-diabetic women for associations of restriction fragment length polymorphism of candidate genes with anthropometric measurements and indices of insulin secretion and resistance. The candidate genes examined were insulin receptor, insulin sensitive glucose transporter and insulin. An association of the class 3 allele of the hypervariable region in the 5' flanking region of the insulin gene was found with upper segment obesity (P = 0.005). Furthermore, the class 3 allele was also associated with fasting hyperinsulinaemia (P = 0.01), stimulated insulin secretion (P = 0.01) and insulin resistance as calculated from the homeostatic model of assessment (HOMA; P = 0.008). No such associations were found with the other candidate genes studied. This data suggests that polymorphisms in the 5' flanking region of the insulin gene may affect expression of the gene and thereby modulate insulin production in severely obese female subjects.


Combination of a delta opioid receptor agonist but not a mu opioid receptor agonist with the D1-selective dopamine receptor agonist SKF 38393 markedly potentiates different behaviors in mice.

The effects of intracerebroventricular injections of opioid peptides selective for mu or delta opioid receptors on behaviors induced by the D1 dopamine agonist SKF 38393 were investigated by using multi-dimensional behavioral analyses. A 10.0 mg/kg dose of SKF 38393 produced a marked increase in grooming behavior. The SKF 38393 (10.0 mg/kg)-induced increase in grooming behavior was clearly antagonized by SCH 23390 (0.03 mg/kg), a D1 dopamine antagonist, but not by S(-)-sulpiride (10.0 mg/kg), a D2 dopamine antagonist. [D-Ala2,MePhe4,Gly5-ol]enkephalin (DAMGO) (0.003 and 0.01 microgram), a mu-selective agonist, or [D-Pen2,L-Pen5]enkephalin (DPLPE) (0.3 or 1.0 microgram), a delta-selective agonist, failed to affect spontaneous behaviors. The combination of DPLPE (0.3 and 1.0 microgram) but not of DAMGO (0.003 and 0.01 microgram) with SKF 38393 (10.0 mg/kg) produced a marked increase in linear locomotion and circuling away from the side receiving the peptide, whereas grooming behavior was not affected. The effects induced by DPLPE (1.0 microgram) plus SKF 38393 (10.0 mg/kg) were fully reversed by the delta-selective opioid antagonist naltrindole (10.0 mg/kg), SCH 23390 (0.03 mg/kg) and S(-)-sulpiride (10.0 mg/kg). These findings suggest that delta but not mu opioid systems interact with D1 dopamine receptors, resulting in a marked increase in linear locomotion and contralateral circuling without causing marked changes in grooming behavior.


An inhibitory effect of isoprenaline on stimulation-induced noradrenaline release from rat atria.

Isoprenaline (0.1 microM), in the presence of phentolamine (1 microM) to block autoinhibitory alpha-adrenoceptors, significantly increased the efflux of radioactivity produced by field stimulation (2 Hz for 60 s) from rat isolated atria in which the noradrenergic transmitter stores had been labelled with [3H]noradrenaline. This facilitatory effect of isoprenaline on noradrenergic transmission was not affected by the selective beta 1-adrenoceptor antagonist CGP 20712A (0.3 microM). However, the selective beta 2-adrenoceptor antagonist ICI 118551 (0.1 microM) not only abolished the facilitatory effect of isoprenaline but reversed it to an inhibitory effect, indicating that the prejunctional beta-adrenoceptors subserving facilitation of noradrenergic transmission in rat atria are of the beta 2-subtype. The inhibitory effect of isoprenaline that was revealed by blockade of beta 2-adrenoceptors was abolished by atenolol (3 microM) in a concentration which markedly reduced the effect of isoprenaline on the rate of atrial beating. This finding suggests that activation of beta 1-adrenoceptors on atrial myocytes by isoprenaline may have resulted in release of one or more substance(s) which inhibited stimulation-induced release of noradrenaline, presumably by activating prejunctional receptors. The inhibitory effect of isoprenaline on noradrenergic transmission was not affected by the prostaglandin synthesis inhibitor indomethacin (10 microM) suggesting that prostaglandins were not involved.


Pharmacology of human dopamine D3 receptor expressed in a mammalian cell line: comparison with D2 receptor.

Two cell lines were created by transfecting cDNAs of the human D2 receptor or the recently cloned human D3 receptor to CHO cells, and the properties of [125I]iodosulpride binding to membranes of these cells were compared. In cell lines expressing the D2 receptor subtype where the selectable marker, a phleomycin-resistance gene, was cotransfected in a different plasmid, a stable expression could be maintained for only few passages. In cell lines expressing the D3 receptor subtype, the selectable marker, a dihydrofolate reductase gene, was cotransfected in the same plasmid and a stable expression could be obtained. In addition, the D3 receptor gene could be amplified in these latter cell lines and a high expression level reached (up to 10(6) binding sites per cell). Sodium and, to a lesser extent, lithium similarly increased [125I]iodosulpride binding to D2 and D3 receptors. In the absence of guanylnucleotide, dopamine had a 24-fold higher apparent affinity at D3 than at D2 receptors. Gpp(NH)p induced rightward shift and steepening of dopamine competition curves at either subtype but the effects were more marked at D2 than at D3 receptors. Several agonists and antagonists, previously regarded as autoreceptor-selective, displayed higher affinities at D3 than at D2 receptors. Although most antagonists used as antipsychotics displayed high affinities at the D3 receptor, all were more potent at the D2 receptor. However, the ratio of Ki values varied over about 10-fold among these compounds, suggesting that they realize differential dopamine receptor subtype occupancy during treatments and that this might be reflected in their clinical profile.


Persistent innervation of the rat neocortex by basal forebrain cholinergic neurons despite the massive reduction of cortical target neurons. II. Neurochemical analysis.

A significant reduction in the activity of cholinergic enzymes in the neocortex is one of the characteristic neurochemical abnormalities in Alzheimer's disease. The withdrawal of cholinergic innervation is thought to be due to atrophy and degeneration of the cholinergic neurons of the basal forebrain, postulated to occur as a consequence of the loss of postsynaptic target neurons and trophic factors synthesized by these target cells. To directly test the dependence of basal forebrain cholinergic innervation on the presence of target neurons, pregnant rats were administered moderate doses of methylazoxymethanol acetate on Gestational Days 14 and 15. This results in a 40-70% reduction of cortical neurons in offspring of treated dams, but has no significant effect on the genesis of basal forebrain cholinergic neurons. In this paper, neurochemical analysis of cholinergic enzymes (ChAT and AchE) in 2-month-old offspring reveals that the basal forebrain innervates the hypocellular cortex in apparently normal fashion, despite the loss of target neurons. The cholinergic innervation of basal forebrain target regions was examined at various times through 20 months of age and was found to retain the initial high levels of enzymatic activity, without any evidence of loss of cholinergic innervation. The preservation of cholinergic innervation over 20 months despite the massive loss of target neurons contradicts the theory of dependence of basal forebrain cholinergic neurons on target-derived trophic support. These results suggest that the degeneration of basal forebrain cholinergic neurons in Alzheimer's disease is due to phenomena more complex than a reduction in the availability of trophic factors.


L1 substrate enhances outgrowth of tyrosine hydroxylase-immunoreactive neurites in mesencephalic cell culture.

We have evaluated neurite outgrowth from mesencephalic tyrosine hydroxylase-positive neurons grown in vitro on different substrates. Cultures of ventral mesencephalon from rat embryos (E13) were plated on plastic dishes coated with the following substrates: L1, L2/HNK-1 "residual" (mainly J1/160 but also tenascin), MAG antigens from mouse brains, laminin, fibronectin, poly-L-lysine, RGD peptide, and plastic alone. After 3, 4, and 6 days in vitro, the cultures were stained using an antibody against tyrosine hydroxylase (TH), and the length of TH-positive neurites was measured by computer-assisted image analysis in a double-blind fashion. L1 antigen had a significant positive effect on neurite outgrowth compared to the other substrates studied. Laminin and fibronectin were also favorable substrates. In cultures treated with cytosine arabinoside to prevent mitoses and glial proliferation, the positive effect of L1 was abolished, but laminin still had a stimulatory effect. These data indicate that L1 may be indirectly involved in differentiation or axonal elongation of substantia nigra dopaminergic neurons and suggest a complex effect involving both neurons and glia on dopaminergic neurite development.


Effect of chronic treatment with (+)-PHNO, a D2 agonist in MPTP-treated monkeys.

A group of four drug naive Macaca fascicularis were rendered parkinsonian with the neurotoxin 1-methyl-4-phenyl-1,2,3,6- tetrahydropyridine and then treated chronically with (+)-PHNO, a potent D2 agonist. After several days, dyskinesia appeared in all animals. At this point, the daily dose of (+)-PHNO was replaced by a dose of the D1 agonist CY 208-243. The substitution by CY 208-243 reproduced the same dyskinesia observed with (+)-PHNO. The administration of the DA synthesis inhibitor AMPT (alpha-methyl-p-tyrosine methyl ester) blocked the dyskinetic and antiparkinsonian effect of (+)-PHNO, and those effects were reestablished by the addition of a subthreshold dose of CY 208-243. Our results show that a selective D2 agonist is capable of inducing dyskinesia and suggest some kind of cooperation between D1 and D2 receptors in the antiparkinsonian and dyskinetic effect produced by (+)-PHNO.


Hypoxia increases extracellular concentrations of excitatory and inhibitory neurotransmitters in subsequently induced seizure: in vivo microdialysis study in the rabbit.

It is uncertain whether a brief hypoxic exposure exerts long lasting effects on central nervous system amino acid neurotransmission. The purpose of this study was to test the hypothesis that a short period of hypoxia would affect release of excitatory and inhibitory amino acids during subsequent bicuculline-induced seizure. Utilizing in vivo microdialysis in cerebral cortex of rabbits, we observed no significant increase in extracellular fluid (ECF) concentrations of the excitatory amino acids, glutamate and aspartate, or the inhibitory amino acids, GABA and taurine, during a 30-min exposure to hypoxia (FiO2 = 0.08). In addition, there was no significant change in these amino acids during uncomplicated seizure. However, when seizure was complicated by a preceding period of hypoxia, there was a marked and progressive rise in both excitatory and inhibitory amino acids in ECF. We conclude that a short period of hypoxia, which itself does not cause changes in ECF concentrations of excitatory amino acids, may nonetheless contribute to neuronal injury by altering the levels of ECF amino acids during a subsequent insult.


Recombinant interleukin-2 (rIL-2) in acquired immune deficiency syndrome (AIDS): preliminary report in patients with lymphoma associated with HIV infection.

In an ongoing phase II study, 12 patients with lymphoma and HIV infection were treated with zidovudine (ZDV) and recombinant interleukin-2 (rIL-2) to evaluate if this association may produce beneficial effect on the immunologic status and the outcome of lymphoma. The protocol included daily doses of rIL-2 at 6 MIU/m2 over 5 days in c.i. per week for a total 4 courses; ZDV was associated at 600 mg/d in the period under study. An improved CD4 count, exceeding 2- to 4-fold the basal count, was obtained in patients with a basal CD4 number greater than 100/microliters accompanied by a significant increase of NK and LAK activity (p less than 0.001). From the clinical point of view the reduction of tumor manifestation was proportional to CD4 basal number; 2 patients from those with CD4 greater than 100/microliters obtained a complete remission after rIL-2 and ZVD. The p24 antigen, taken as parameter of viral replication, remained invariably negative after rIL-2 and ZDV in patients already negative and became negative in 1 patient previously positive. Our conclusion is that the association of rIL-2 and AZT is safe and useful in patients with lymphoma and HIV infection.


Essential catalytic role of Glu134 in endo-beta-1,3-1,4-D-glucan 4-glucanohydrolase from B. licheniformis as determined by site-directed mutagenesis.

Site-directed mutagenesis experiments designed to identify the active site of Bacillus licheniformis endo-beta-1,3-1,4-D-glucan 4-glucanohydrolase (beta-glucanase) have been performed. Putative catalytic residues were chosen on the basis of sequence similarity analysis to viral and eukaryotic lysozymes. Four mutant enzymes were expressed and purified from recombinant E. coli and their kinetics analysed with barley beta-glucan. Replacement of Glu134 by Gln produced a mutant (E134Q) that retains less than 0.3% of the wild-type activity. The other mutants, D133N, E160Q and D179N, are active but show different kinetic parameters relative to wild-type indicative of their participation in substrate binding and transition-state complex stabilization. Glu134 is essential for activity; it is comprised in a region of high sequence similarity to the active site of T4 lysozyme and matches the position of the general acid catalyst. These results strongly support a lysozyme-like mechanism for this family of Bacillus beta-glucan hydrolases with Glu134 being the essential acid catalyst.


A novel specific binding site for anxiolytic homophthalazines in the rat brain.

Radioligand binding studies were performed in order to elucidate the mechanism of action of anxiolytic-neuroleptic homophthalazines. Rat striatal membrane preparations were found to bind 3H-EGIS 6775 [3H-GYKI-52 322, 3H-(1-(4-aminophenyl)-4-methyl-7,8-dimethoxy-5H-homophthalazine)] in a specific and displaceable manner. Several other brain regions tested were devoid of similar binding activity. Saturation analysis revealed that binding affinity was in the 10(-8)-10(-7) M range. Binding was enhanced by Mg2+ ions and, to a smaller extent by Ca2+ ions. The binding principle was sensitive to heat or trypsin treatment. This specific binding site appears, according to competition studies, different from the receptors whose presence in the rat striatum has been reported earlier.


Maternal mortality: community-based interventions.

Maternal mortality is one of the great neglected problems of health care in developing countries. The World Health Organization estimates that approximately 500,000 women die each year from pregnancy-related causes, over 98% of these deaths occurring in the developing world, where maternal mortality is as much as 100 times higher than rates seen in industrialized countries. The most common causes include obstructed labor and ruptured uterus, postpartum hemorrhage, eclampsia, postpartum infection and complications of illegal abortion. It is suggested that no new or costly technologies are needed; rather that appropriate priority setting and allocation of needed resources are essential to the solution of the problem. There are few interventions that hold much hope of success at the village level, although antibiotics, ergotrate, and sedatives might be productively utilized, after appropriate training. Overall, however, networks of maternity care facilities, trained personnel, and means of transport are necessary to provide needed emergency maternity care services.


General principles in delegation of maternity functions to community-based health workers.

Following a review of the general principles in delegation of responsibility, the functions to be delegated are presented together with minimal criteria for training to acquire required skills. Problems arising from delegation are reviewed as well as possible solutions which include modification of some standard procedures. It is concluded, drawing from case summaries, that effective delegation needs strong leadership of maternal health teams buttressed with national political, professional, and community support.


Rural maternity care in Sierra Leone.

The training and duties of a new category of health worker, the Maternal and Child Health Aide, is described. She is a literate women recruited from the place where she will eventually work. She has become the immediate supervisor of the traditional birth attendants (TBAs), who continue to conduct most deliveries in Sierra Leone.


[Effect of various neurotropic drugs on chronic hyperreflexia in rats after cordotomy].

Substances modulating calcium permeability of cell membrane (verapamil imidazol, 4-aminopyridine), decreasing motor activity (meprobamate) and blocking adrenoreceptors (clophelin, propranolol, droperidol, aminazine++ have been studied for their action on the monosynaptic discharge of the ventral roots (MD VR) reinforced due to chronic cutting of the spinal cord. It is found that verapamil and meprobamate depress more strongly MD VR of rats with chronic cutting of the spinal cord than of those with the acute one. Imidazol and 4-aminopyridine reinforcing MD VR of rats with acute cutting of the spinal cord have no influence on the analogous index of rats with chronic cutting of the spinal cord. Adrenoblockers do not change the amplitude of MD VR in both groups of the animals.


Delegation of responsibility in maternity care in Karawa rural health zone, Zaire.

Karawa Health Zone (KHZ) located in northwestern Zaire, comprises 19,000 km2 with about 340,000 inhabitants. As part of an expanding primary health care program, KHZ officials instituted an extensive outreach program to increase access of women to maternity care. General practitioner (GP) doctors, nurses, midwives, auxiliaries, and traditional birth attendants share responsibilities in delivering maternity care services. Nurses and midwives perform most of the obstetric functions at the Karawa Hospital Maternity Center. Data from 1988 to 1990 show that nurses and midwives attended 98% of normal deliveries and 81% of the complicated ones. Selected nurses performed cesarean section routinely. There was no significant difference in maternal and neonatal outcomes for cesarean section performed by nurses and GPs. Physicians on staff provided support through monitoring, supervision, training, and care for the most complicated cases.


Mozambique--delegation of responsibility in the area of maternal care.

The shortage of physicians in Mozambique has necessitated the training and delegation of maternity care to other available health personnel, such as traditional birth attendants, midwives, and nurses. In addition, surgical technicians have been trained to perform major surgical procedures, including cesarean sections and hysterectomies.


Life-saving skills training for midwives: report on the Ghanaian experience.

The shortage of rural-based physicians in Ghana has led to a decision to provide short term courses to rural midwives on the treatment of those obstetric conditions that are the main causes of maternal mortality. A description of this ongoing training is provided.


Delegation of obstetric care in Indonesia.

An intervention and a control area were selected. Results of the intervention, which consisted of training TBAs, midwives, and health center doctors, were positive inasmuch as there was an increase in referrals of breech presentation, prolonged labor and intrapartum fever in the intervention area. Pregnant women in general were also more often referred in the intervention area as compared to the control area. The same outcome was seen in referrals of neonates weighing under 2000 or more than 4000 g. At the district hospital level, a great number of procedures were left to general practitioners with quite satisfactory results. Conclusions drawn from our experience are that: (1) obstetric care must of necessity be delegated to TBAs, midwives, and general practitioners; (2) TBAs can be taught to recognize danger signs; (3) general practitioners are able to do such procedures as curettage, vacuum or forceps deliveries, and manual removal of placenta.


Appropriate training for maternal health and the attributes of the trainer.

This review describes the qualities which those who are trainers in safe motherhood and other programs should display. The educational principles and objectives for maternal health workers, based on the tasks which they have to do and expressed in the skills and competences they must attain, are related to practical methods of training. Problem-based learning is advocated and the value of distance education for rural workers is discussed.


Traditional birth attendant training: sharing experiences.

The experiences in TBA training in five countries are contrasted. Successful training must impart improved skills to the TBAs and follow-up that both provides continuing support and meets the self-interest of the TBAs. The preference of mothers for services of TBAs is explained.


Criteria for the selection of experiences and content in midwifery education.

Selection of content and learning experiences for any educational endeavor is based on the aim and objectives for setting up the program, and the competencies required of graduates of the school. To exclude those experiences that will lead to skill in the management of obstetric emergencies, and at the same time expect midwives to function independently in rural communities is a negation of basic curriculum planning principles. The range of experiences available to the contemporary curriculum planner in the health field is infinite. However, due to the changing nature of societal needs in the health field, and the rapid obsolescence of previously held assumptions about health care services and the role of midwives, selection of experiences requires creativity and well defined criteria for judgment. Those suggested in this paper are validity, comprehensiveness, pattern, balance, relevance, and continuity. The six criteria are discussed to show the relevance of each to some of the current weaknesses in midwifery education in Nigeria.


Alternative model for low risk obstetric care in Third World rural and peri-urban areas.

The Program for Integrated Health Care (PROAIS) has devised ways and means of reaching the traditional birth attendant, here called empiric midwife and training her to offer adequate, hygienic care at delivery for her low-risk patients. She is taught to recognize the high-risk patient and to send her to the weekly prenatal clinics staffed by physicians and registered nurses. The empiric midwife can also be taught prenatal care, detection of cervical and breast cancer, family planning, and the importance of breastfeeding. These women are considered colleagues by their formal medical counterparts and are treated as such. More recently some of the more capable have been placed on County Health Department payrolls.


Evaluation of maternal health programs: approaches, methods and indicators.

Evaluation of maternal health services is an essential function of any health system. Single or multiple aspects of a program can be evaluated but they must be clearly defined. To assess success of a program the dimensions of coverage, equity, quality, women's satisfaction, efficiency and cost-effectiveness must be considered. These can be measured according to some standard criteria, and various methods are described, including randomized clinical trials, randomized community trials, before and after studies, and observational studies, including the case-control approach and confidential inquiries into maternal deaths. Indicators to assess effectiveness of a maternal health program may relate to structure, i.e., available resources or organizational arrangement; process, including changes in quantity of services provided; or outcome indicators that measure maternal mortality or morbidity.


Midwives: key rural health workers in maternity care.

The most acceptable and attainable rural health worker for maternity care is frequently the traditional birth attendant or other personnel lacking clinical skills to treat life-threatening emergencies. When first referral level facilities are also poorly staffed and ill-equipped to deal with these emergencies, this again points to the need for training of and delegation to the trained midwife in rural areas. Unfortunately, their number is declining in rural areas of some countries most in need, e.g., Tanzania. Elsewhere, midwifery skills and knowledge have been integrated into basic nursing education, but practical skills are only developed postbasically when midwife educators are expert clinicians. The graduates of such training could be delegated responsibility for many lifesaving procedures in obstetric care. Successful clinical experience in use of these responsibilities will earn the midwife's needed community reputation as a trusted health worker.


Somatostatin is an essential paracrine link in acid inhibition of gastrin secretion.

We studied the functional coupling between antral somatostatin and gastrin cells in pigs using isolated perfused preparations of the antrum with intact supply of the vagus nerves. Luminal acidification significantly inhibited gastrin secretion to 61 +/- 3% of basal secretion and increased somatostatin output 9-fold. Intra-arterial infusion of somatostatin to concentrations of 10(-10) and 10(-9) mol/l inhibited gastrin secretion to 18 +/- 9 and 33 +/- 11% of basal secretion. Electrical stimulation of the vagus nerves and intra-arterial infusion of gastrin-releasing polypeptide (GRP; 10(-9) mol/l) significantly increased both gastrin and somatostatin secretion. Addition to the perfusate of Fab fragments of somatostatin antibodies abolished the effect of somatostatin at 10(-10) mol/l and the acid inhibition of gastrin secretion, but had no effect on the response to vagus stimulation of GRP infusion. We conclude that a local release of somatostatin is essential for the acid-induced inhibition of gastrin secretion, whereas changes in the local somatostatin concentration are unlikely to play a major role in vagally or GRP-induced gastrin secretion.


Expression of a zebrafish caudal homeobox gene correlates with the establishment of posterior cell lineages at gastrulation.

This paper deals with the first identification of a caudal cDNA containing a homeobox of the Drosophila caudal family in the zebrafish. A cDNA library from late gastrula stage embryos was constructed and screened with a mouse Cdx 1 homeobox probe. A 1.6 kb cDNA clone containing a homeobox related to other caudal homeoboxes was isolated and called cdx[Zf-cad1]. Analysis of the predicted 301 amino acid translation product reveals additional regions of homology outside the homeodomain with other members of the caudal family. Particularly, the cdx[Zf-cad1] putative protein shares a conserved N-terminal region with its chicken homolog CHox-cad. Transcripts are first detected just before the onset of gastrulation. At the beginning of gastrulation, a single 1.8 kb cdx[Zf-cad1] transcript is located near the blastoderm margin with a high level of expression restricted to the epiblast. At this stage, the hypoblast is clearly negative. At the end of gastrulation, cdx[Zf-cad1] is widely expressed in vegetal (i.e. prospective posterior) epiblast and hypoblast, with a somewhat weaker expression in the dorsal hypoblast. During somitogenesis, cdx[Zf-cad1] exhibits a posterior regionalization in the neurectoderm. In contrast, no expression is detected in the mesoderm of 22 h embryos (late somitogenesis). Posterior endoderm is also positive at this stage. cdx[Zf-cad1] transcripts cease to be detected about 48 h after fertilization. They are undetectable in the adult, particularly in female gonads. The pattern of cdx[Zf-cad1] expression during and after gastrulation is consistent with its possible involvement in the regionalization of the embryo at these stages.(ABSTRACT TRUNCATED AT 250 WORDS)


HLA class II molecules and autoimmune hepatitis susceptibility in Japanese patients.

To investigate the association between autoimmune hepatitis and HLA alleles in Japanese patients, serological typing and class II genotyping were performed using the polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) method. Serological typing showed that HLA-B54, -DR4, -DR53, and -DQ4 were significantly more frequent in patients with autoimmune hepatitis than in controls. HLA-DR4 was most frequently associated with autoimmune hepatitis (88.7%). In PCR-RFLP typing, the frequency of DRB1*0405 was significantly higher in autoimmune hepatitis than in controls. However, there was no significant difference in the frequency of Dw between the patients and the controls who were DR4-positive. The significant increase observed in DQA1*0301 and DQB1*0401 was explained by a linkage disequilibrium with DR4. Six DR4-negative patients had DR2, but there was no significant difference in the frequency of the DR2-associated Dw-alleles compared with the DR2-positive controls. No DPB1 allele was significantly associated with autoimmune hepatitis. These findings suggest that the basic amino acid at position 13, which is present only on the DR2 and DR4 B1 molecules (Arg on DR2 and His on DR4), contributes to the susceptibility to autoimmune hepatitis among the Japanese.


Rapid identification of bacteria of the Comamonadaceae with amplified ribosomal DNA-restriction analysis (ARDRA).

Ribosomal rRNA gene fragments (rDNA) encompassing the 16S rDNA, the 16S-23S rDNA spacer region and part of the 23S rDNA of 95 strains belonging to 13 well-described taxa of the eubacterial family Comamonadaceae (beta subclass of the Proteobacteria or rRNA superfamily III) were enzymatically amplified using conserved primers. The fragments of approximately 2400 base pairs were subjected to restriction analysis. Restriction fragment length patterns obtained with HinfI enabled us to distinguish 9 of the 13 taxa studied. Restriction with CfoI was necessary to differentiate Acidovorax delafieldii from A. temperans and Hydrogenophaga flava from H. pseudoflava. The results indicate that amplified rDNA restriction analysis is a simple and reliable tool for the identification of bacterial species.


Characterization of Serpulina (Treponema) hyodysenteriae and related intestinal spirochetes by ribosomal RNA gene restriction patterns.

Serpulina (Treponema) hyodysenteriae strain A-1 partially purified rRNA, labelled with photobiotin, was used as a non-radioactive probe to identify the rRNA gene restriction patterns of S. hyodysenteriae strains and other spirochetes. Sau3A restriction enzyme digests resulted in similar rRNA gene restriction patterns in S. hyodysenteriae strains from five different countries. Some S. hyodysenteriae strains could be differentiated by variations in their rRNA gene restriction patterns after cleavage of DNA by restriction enzymes SspI or BglII. S. innocens and Treponema succinifaciens, non-pathogenic pig intestinal spirochetes, had rRNA gene restriction patterns that differed markedly from the S. hyodysenteriae patterns, and from each other.


[Elevated serum levels of ICAM-1 in Wegener's granulomatosis].

Circulating soluble ICAM-1 (sICAM-1) isoforms are known to be elevated in inflammatory and autoimmune disorders (SLE, RA). A correlation between levels of circulating sICAM-1 and disease activity was described (5). In sera from 31 patients suffering from histologically confirmed Wegener's granulomatosis (WG) elevated levels of ICAM-1 were now detected in this primary systemic vasculitis. In generalized active disease circulating ICAM-1 levels were significantly higher when compared to remission and healthy control individuals. Sera from WG patients collected during intercurrent infections while on immunosuppressive drugs showed no elevation of circulating ICAM-1.


[Enhanced lymphocyte proliferation in the presence of epidermal cells of HIV-infected patients in vitro].

Clinical observations show that the HIV infection is often associated with affections of the skin. In order to examine the involvement of the epidermal immune system in the HIV infection, we determined accessory cell function of epidermal cells from HIV-1-infected patients. For this we measured the proliferative response of enriched CD(4+)-T-lymphocytes from HIV-infected patients and noninfected controls to stimulation with anti-CD3 and IL-2 in the presence of epidermal cells; the enhancement of the response is dependent on the presence of functionally intact accessory cells. The capacity of epidermal cells to increase the anti-CD3-stimulated T-cell proliferative response was significantly enhanced in HIV patients (CDC III/IVA) as compared with noninfected donors. It is discussed, whether the increased activity of epidermal cells from HIV-infected patients may be responsible for several of the dermal lesions in the course of an HIV infection as due to an enhanced production and release of epidermal cell-derived cytokines.


Presence of functional receptors for the Escherichia coli heat-stable enterotoxin in the gastrointestinal tract of the chicken.

Receptors for the Escherichia coli heat-stable enterotoxin (STa) were shown to be present throughout the digestive tract of the chicken, with binding activity present not only in the intestinal epithelium but also in the intestinal smooth muscle. Brush border membrane vesicles (BBMV) purified from chicken enterocyte homogenates and plasma membranes (SMPM) purified from intestinal smooth muscle homogenates were compared with pig enterocyte BBMV. All had similar 125I-STa binding affinities, but the 50% effective concentration for STa activation of guanylate cyclase was higher in SMPM than in BBMV. Maximal STa-stimulated guanylate cyclase activities were similar in chicken and pig BBMV and were seven- to eightfold higher than in SMPM, and the STa receptor density was five- to sixfold higher. Patterns unique to each membrane were demonstrated after affinity labelling of STa receptors with 125I-STa, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and autoradiography. The results demonstrated STa-stimulated guanylate cyclase activity in birds as well as mammals and suggested that there are different functional STa receptors in chicken BBMV and SMPM.


Genetic relationship of putative colonization factor O166 to colonization factor antigen I and coli surface antigen 4 of enterotoxigenic Escherichia coli.

Plasmid DNA from two strains of enterotoxigenic Escherichia coli harboring genes encoding coli surface antigen 4 (CS4) and from seven Indian enterotoxigenic E. coli isolates cross-hybridized at low stringency but not at high stringency with two polynucleotide probes derived from the colonization factor antigen I (CFA/I) operon. Low-stringency Southern blot hybridization of PstI-digested plasmid DNA from the seven Indian isolates yielded characteristic restriction fragment patterns, distinct from those of CS4- and CFA/I-associated plasmid DNA. Two of the Indian strains were transformed with a recombinant plasmid harboring the cfaD gene, which encodes a positive regulator of CFA/I and CS4 genes. The cfaD transformants produced large amounts of putative colonization factor O166 (PCFO166) irrespective of whether the nutrient agar contained bile salts, a growth factor otherwise required for adequate PCFO166 expression. A considerable interstrain variation in the level of PCFO166 production could be explained by differences in the proportion of bacteria that were fimbriated, as visualized by electron microscopy. The N-terminal amino acid sequence of PCFO166 fimbrial protein showed a high degree of homology with the corresponding sequences of CFA/I and CS4.


Immune CD4+ T cells specific for Theileria parva-infected lymphocytes recognize a 24-kilodalton protein.

Theileria parva is a protozoan parasite that infects and transforms bovine lymphocytes. Here we report the partial purification of a T. parva-specific protein from infected lymphocytes that is recognized by CD4+ parasite-specific T-cell clones derived from immune cattle. T. parva-infected lymphocytes were homogenized in Dulbecco's phosphate-buffered saline in the presence of protease inhibitors. The antigen was purified from a postmicrosomal supernatant by using a combination of DEAE-cellulose chromatography and hydroxylapatite column chromatography. After labelling with 125I, the antigen preparation was subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and found to contain 8 to 10 proteins. This preparation was subjected to chromatography in phosphate-buffered saline on HPLC TSK-250/125 columns coupled in tandem. A radiolabelled protein of M(r) 24,000 correlated with antigenic activity.


P-glycoprotein transports corticosterone and is photoaffinity-labeled by the steroid.

Multi-drug-resistant cells overproduce a 130-180-kDa integral membrane phosphoglycoprotein known as P-glycoprotein which acts as an energy-dependent drug efflux pump. While P-glycoprotein has been shown to transport hydrophobic anti-tumor drugs out of multi-drug-resistant cells in tissue culture, its endogenous substrates remain unknown. This report shows that 3H-corticosterone can specifically photoaffinity label P-glycoprotein. Furthermore, corticosterone is effluxed from multi-drug-resistant cells by P-glycoprotein. These data suggest that corticosterone may be an endogenous substrate for P-glycoprotein.


Differentiation-related expression of adhesion molecules and receptors on human neuroblastoma tissues, cell lines and variants.

The expression of cellular adhesion molecules (CAM) involved in cell adhesion and immune recognition was measured on neuroblastoma tissue samples, on a neuroblastoma (NB) cell line, SK-N-SH, and on 3 phenotypically different variants, SH-SY5Y, SH-EP, SH-IN, representing neuronal, Schwannian/glial or intermediate NB-cell types. Immunohistochemical analysis of CAM expression by NB and related tumors at different stages of differentiation revealed a co-expression of several CAM (ICAM-1/CD54, LFA-3, VLA-2 and HLA-ABC) associated with low stages and more highly differentiated NB tumors and peripheral neuroepitheliomas (PN). In contrast, N-CAM was uniformly expressed on all NB tumors. Flow cytometric analysis of CAM surface expression by SK-N-SH and variant cells revealed highly variable phenotypes. Expression of ICAM-1, LFA-3, VLA-2 and HLA-ABC molecules was associated with the epithelial cell type represented by the SH-EP variant. In contrast, low expression of these molecules and high expression of N-CAM was associated with the neuronal SH-SY5Y cells. Exposure of the NB cells to differentiation inducers (retinoic acid, 5'-bromodeoxyuridine and phorbol esters) and cytokines (tau-interferon, alpha-tumor necrosis factor) resulted in a variable up-regulation of the expression of all CAMs, except N-CAM, regardless of the type of differentiation induced. In an attempt to establish a link between the pattern of expression of CAM on NB cells and their susceptibility to natural killer (NK) or lymphokine-activated killer (LAK) cell lysis, the analysis revealed that NB cells expressing CAM and a differentiated phenotype were less susceptible to NK lysis, but no difference in the sensitivity of the NB cell types to LAK effectors was observed. Treatment of NB target cells with cytokines or PMA decreased their susceptibility to NK and LAK lysis, while induction of differentiation with RA or BUdR resulted in no changes in the sensitivity to NK and LAK lysis. In conclusion, expression of HLA-ABC and several co-regulated CAMs was shown to be associated with a differentiated phenotype in NB, with an overall decreased sensitivity to NK/LAK effector cells.


Human urinary glycosaminoglycans as accurate method for ovulation detection.

Urinary glycosaminoglycans (GAGs) content showed a characteristic pattern of fluctuation during normal and hormonally induced cycles with a distinct peak at ovulation. This peak of maximal GAGs concentration (106.7 +/- 46.2 micrograms/mL in urine) was centered according to the day of the midcycle LH surge, which serves as the reference point, designated day 0. All cycles are presumed to be ovulatory on the basis of biphasic BBT nadir, ultrasonographic studies, and progesterone assays (greater than 5 ng/mL of serum) during the secretory phase. In hormonally induced menstrual cycles, a noticeable increase in GAGs concentration (70%) was observed. This indirect evidence suggests a possible correlation between GAGs content and hormonal effect. All types of GAGs, with the exception of heparin, have been found in urine, chondroitin sulfate C being the major component at time of ovulation. These results strongly suggest that urinary GAGs determination is a precise method for ovulation detection.


Reproductive outcome following two ectopic gestations: results after conservative surgery.

In an 11-year period, 106 women were treated for two ectopic pregnancies (EP). Thirty-one had been surgically sterilized by the second operation. Of the 75 women with at least one patent tube, 70 (93%) were followed up regarding pregnancy outcome, with a mean follow-up period of 4.5 years. Fifty women had had a desire for pregnancy after treatment for the second EP. Of these, 26 (52%) did not achieve any conception; 16 (32%) experienced at least one more EP, and 13 (26%) had at least one intrauterine pregnancy. There were a total of 46 pregnancies, of which 21 (46%) were ectopic and 16 (35%) were carried on term. When conservative surgical techniques had been performed at the first EP, the number of intrauterine pregnancies achieved after the second EP was doubled, and the risk for a third EP was not raised. There was found no statistical difference in laterality of the second EP when the first operation was conservative. It is concluded that women experiencing repeat EP have severely impaired future fertility, with a high risk of a third EP if they conceive.


Comparison of intrauterine insemination, intracervical insemination, and timed intercourse in women treated with human menopausal gonadotropin.

The effects of intrauterine insemination (IUI), intracervical insemination (ICI), and timed intercourse (TI) in women who were treated with human menopausal gonadotropin (hMG) for anovulation were evaluated. The pregnancy rates per cycle following IUI (70 cycles), ICI (62 cycles) and TI (158 cycles) were 20%, 9.6%, and 17.7%, respectively; these differences are not statistically significant. The abortion rate and the multiple pregnancy rate were also not significantly different. This report suggests that in women who are undergoing treatment with hMG, there is no added benefit from artificial insemination (either intrauterine or intracervical insemination) over timed intercourse.


On the relationship between endocrine and ovulatory abnormalities, and polycystic ovaries as diagnosed by ultrasonography.

The present study was undertaken to investigate the relationship between serum testosterone levels, the menstrual cycle, and polycystic ovaries as shown by ultrasonographic methods. The patients were divided into four groups: (1) cases with normal-appearing ovaries and with regular menstruation (control); (2) cases with normal ovaries on ultrasound, but with abnormalities of the menstrual cycle (MA); (3) cases with polycystic ovaries, but normal menstrual cycle (PCO); (4) cases with both polycystic ovaries and menstrual abnormalities (PCOMA). The results were as follows. (1) The ovarian area was significantly larger in the PCOMA group than in the control group. (2) The number of cases with elevated LH (greater than 30 mIU/mL) was significantly higher in the PCOMA group than in the other groups. (3) In comparison with controls, the percentage of cases with LH/FSH ratios greater than 3.0 was significantly higher in all three groups. (4) The estrone/17 beta-estradiol ratio for the PCOMA group was significantly greater than those of the other groups. (5) The testosterone levels of the PCO and PCOMA group were significantly greater than the control value. (6) The percentage of endocrine criteria (LH greater than 30 mIU/mL or LH/FSH ratio greater than 3.0, estrone/17 beta estradiol ratio greater than 1.0 testosterone greater than 75, greater than 100, and greater than 125 ng/dL) for PCOMA was significantly higher than that of the MA group and the control group, but not significantly greater than the PCO group. It is suggested that morphological changes in the ovaries show a closer correlation with abnormalities of endocrine function than does the presence or absence of abnormalities of the menstrual cycle.


Cervical cerclage in uterine malformations.

This retrospective study represents our experience with cervical cerclage (suture) in pregnancies with uterine malformation. Seventeen patients with uterine malformations were involved. In these patients, the outcome of 31 gestations with cervical cerclage was evaluated. Uterine malformation was associated with cervical incompetence in 41% of the patients. Term delivery rate and fetal loss were observed in 56% and 26% of pregnancies, respectively. Infant salvage was not significantly different regardless of whether cervical incompetence was present or absent. A prolonged hospital stay and frequent use of tocolysis were noted during gestations with cerclage. Our data suggest that the outcome of pregnancies with uterine malformation was not improved by cervical cerclage when the indication for cerclage was the malformation itself.


HLA gene analysis in a Japanese family with hypertrophic cardiomyopathy by restriction fragment length polymorphism.

A large Japanese family with hypertrophic cardiomyopathy (HCM) was examined (n = 61). Ten of 14 affected family members who showed HCM on the basis of electrocardiography and two-dimensional echocardiography were subjected to human leukocyte antigen (HL-A)-A,B,C typing by the lymphocyte cytotoxicity test and D, DR typing by restriction fragment length polymorphism (RFLP). As control, the HLA genotype of 14 non-affected family members was analyzed. HLA typing of the affected subjects with HCM revealed a very close association (67%) of HLA-DR (4w8) among A,B,C,D and DR. However, the LOD scores of HLA-DR4 and -DR w8 were 0.693 (theta = 0.35) and 0.642 (theta = 0.30) respectively. These data suggest that HLA-loci, analyzed on the basis of RFLP, may not be related with familial HCM with or without obstruction.


Morphological and quantitative study of multinucleated bodies appearing in rat seminiferous tubules after bilateral caput epididymectomy.

Adult Wistar rats were bilaterally caput epididymectomized and the effects on testicular germinal epithelium and formation of multinucleated bodies were studied and quantified at 2, 4, 7, 14, 21, and 28 days after surgery. Sham-operated and bilaterally efferentectomized animals served as controls. No alterations were found in sham-operated animals. Efferentectomized animals showed a progressive alteration of the seminiferous tubule epithelium and a (very occasional) presence of multinucleated bodies. Epididymectomized animals presented a progressive degeneration of the germinal epithelium, which was almost complete at 28 days. This epithelial degeneration was accompanied by the formation of multinucleated bodies from germinal cells, whose number and characteristics varied with the experimental interval. The multinucleated bodies described here resemble the multinucleated cells mentioned by other authors. They do not seem to be cellular; instead, they appear to be debris, since electron microscopic observations do not reveal a plasma membrane.