The presence of a dehydroepiandrosterone-specific receptor binding complex in murine T cells.

We have investigated the ability of dehydroepiandrosterone (DHEA) to alter the production of interleukin-2 (IL-2) and to bind to a specific binding complex in antiCD3 epsilon activated T cells. Binding activity correlated with the presence of a specific DHEA binding complex in the cytosol and nuclei of DHEA-responsive T-cell hybridomas, as well as in CD4+ and CD8+ cells isolated from peripheral lymph nodes of normal mice. Scatchard analysis determined that intact lymphocytes and cytosolic fractions contained high affinity binding for [3H]DHEA (approx. 2.6 nM) with 1000-7000 binding sites existing per cell. Five of the T-cell hybridomas tested both responded to DHEA treatment with increased production of IL-2 and also contained specific high affinity [3H]DHEA binding. Four additional T-cell hybridomas were found to contain no specific [3H]DHEA binding and were also unresponsive to DHEA influences on IL-2 production. Sucrose density gradients demonstrated a 3-4s [3H]DHEA binding complex in high salt and a 7-8s binding complex in low salt. Specific binding was inhibited by preincubation of the cytosol fractions with either trypsin or chymotrypsin, or by heating to 60 degrees C for 1 h (less than 15% of control). [3H]DHEA binding was unaffected by preincubation of the cytosol fractions with ribonuclease, deoxyribonuclease, or phospholipase A. The DHEA-protein complexes bound to DNA-cellulose with the amount of binding being slightly increased by preincubation at 25 degrees C as compared to 4 degrees C. As expected, [3H]DHEA binding was inhibited by the addition of unlabeled DHEA, but was also modestly inhibited by dihydrotestosterone and cortisol. Binding of DHEA was unaffected by progesterone, dexamethasone, estradiol, androsterone, DHEAS, and beta-etiocholanolone at all concentrations tested. DHEA was incapable of inhibiting the binding of [3H]DHT to the androgen receptor or [3H]dexamethasone to the glucocorticoid receptor. Collectively, these findings suggest that murine T cells contain a specific DHEA receptor. We believe that DHEA is a steroid hormone that is directly involved in the regulation of IL-2 production by both normal and some T-cell hybridomas.


Resonance Raman microprobe spectroscopy of rhodopsin mutants: effect of substitutions in the third transmembrane helix.

A microprobe system has been developed that can record Raman spectra from as little as 2 microL of solution containing only micrograms of biological pigments. The apparatus consists of a liquid nitrogen (l-N2)-cooled cold stage, an epi-illumination microscope, and a substractive-dispersion, double spectrograph coupled to a l-N2-cooled CCD detector. Experiments were performed on native bovine rhodopsin, rhodopsin expressed in COS cells, and four rhodopsin mutants: Glu134 replaced by Gln (E134Q), Glu122 replaced by Gln (E122Q), and Glu113 replaced by Gln (E113Q) or Ala (E113A). Resonance Raman spectra of photostationary steady-state mixtures of 11-cis-rhodopsin, 9-cis-isorhodopsin, and all-trans-bathorhodopsin at 77 K were recorded. The Raman spectra of E134Q and the wild-type are the same, indicating that Glu134 is not located near the chromophore. Substitution at Glu122 also does not affect the C = NH stretching vibration of the chromophore. The fingerprint and Schiff base regions of the Raman spectra of the 380-nm, pH 7 forms of E113Q and E113A are characteristic of unprotonated retinal Schiff bases. The C = NH modes of the approximately 500-nm, pH 5 forms of E113Q and E113A in H2O (D2O) are found at 1648 (1629) and 1645 (1630) cm-1, respectively. These frequencies indicate that the protonated Schiff base interacts more weakly with its protein counterion in the Glu113 mutants than it does in the native pigment. Furthermore, perturbations of the unique bathorhodopsin hydrogen out-of-plane (HOOP) vibrations in E113Q and E113A indicate that the strength of the protein perturbation near C12 is weakened compared to that in native bathorhodopsin.(ABSTRACT TRUNCATED AT 250 WORDS)


Incorporation of dietary oleate, linoleate, alpha-linolenate and eicosapentaenoate into the plasma lipid fractions of four strains of rat.

Four strains of rat (Dark Agouti, DA; Ginger Hooded, GH; Portion, P; Hooded Wistar, HW) were fed elemental diets containing different sources of fat at the 10% (w/w) level. The dietary fats used included the following oils; olive (rich in oleate), sunflower (rich in linoleate), linseed (rich in alpha-linolenate) and fish (rich in eicosapentaenoate and docosahexaenoate). Differences between strains in response to individual diets were modest compared with the much greater differences achieved by the dietary treatments. In general, the changes in polyunsaturated fatty acid (PUFA) levels in the plasma lipids of all rat strains followed the major PUFA in the diet. There were, however, strong interactions between dietary n-6 and n-3 PUFA which affected not only the level of particular PUFA in lipid fractions but also the lipid fraction in which a particular PUFA appeared. Our findings indicate that a response to dietary fats in the plasma lipids of one strain of rat can be expected to occur with relatively minor variations in other commonly used rat strains.


Characterization of putative neurotransmitter N-acetyl-aspartyl-glutamic acid and some related compounds by fast atom bombardment and tandem mass spectrometry.

The characterization of the N-blocked neuropeptide N-acetyl-aspartyl-glutamic acid and of some related compounds was carried out by negative-ion fast atom bombardment mass spectrometry and collision-induced dissociation of the generated [M-H]- species. Thorough analysis of the fragment ion spectra allowed discrimination between sequence isomeric compounds, and highlighting of differences between dipeptides linked in the normal or iso mode.


Depressive syndromes in schizophrenic patients under neuroleptic therapy. ANI Study Group Berlin, Düsseldorf, Göttingen, Munich, Federal Republic of Germany.

Schizophrenic outpatients (= 364) were assigned at random to three different treatment strategies: (1) continuous medication with neuroleptic drugs, (2) intermittent medication with crisis intervention and (3) intermittent medication with early intervention. Depressive syndromes were rated according to three different scales for depressive syndromes (Brief Psychiatric Rating Scale anxious depression factor, Arbeitsgemeinschaft für Methodik und Dokumentation in der Psychiatrie/depression, and the self-rating Paranoid Depression Scale) after 1 and 2 years of treatment. No differences in depression scores were found between the three treatment strategies. Comparisons between patients treated with neuroleptic drugs at the time and patients without neuroleptics revealed significantly higher depression scores in the neuroleptics group in most comparisons. No differences were found between patients treated with low versus high potency neuroleptics and between oral versus depot neuroleptics. However, depression correlated with extrapyramidal symptoms.


The inheritance of type I and type III von Willebrand's disease in Israel: linkage analysis, carrier detection and prenatal diagnosis using three intragenic restriction fragment length polymorphisms.

Three intragenic restriction fragment length polymorphisms (RFLPs) were used to study linkage and analyse the mode of inheritance in type I and type III von Willebrand's disease (vWD). In two families linkage was established between Sac I RFLPs and the inheritance of type I vWD. RFLP analysis of amniocyte DNA from a potentially affected foetus enabled us to establish a prenatal diagnosis of vWD in a third family with type I vWD. Linkage was also established in four families between the Sac I and two Taq I RFLPs and the inheritance of type III vWD. All type III probands were homozygotes and inherited the same mutant vWF allele from both parents. Heterozygous carriers from one type III family were phenotypically normal and could be detected only by linkage analysis, whereas carriers from the remaining three type III families were asymptomatic but had decreased values of vWF antigen and activity. RFLP-based linkage analysis of vWD alleles provides a way to improve the diagnostic precision, detect carriers, and may be useful for prenatal diagnosis of type III vWD.


Immunocytochemistry and calcium cytochemistry of the mammalian pineal organ: a comparison with retina and submammalian pineal organs.

Morphologically the mammalian pineal organ is a part of the diencephalon. It represents a neural tissue histologically ("pineal nervous tissue") and is dissimilar to endocrine glands. Submammalian pinealocytes resemble the photoreceptor cells of the retina, and some of their cytologic characteristics are preserved in the mammalian pinealocytes together with compounds demonstrable by cyto- and immunocytochemistry and participating in photochemical transduction. In our opinion, the main trend of today's literature on pineal functions--only considering the organ as a common endocrine gland--deviates from this structural and histochemical basis. In mammals, similar to the lower vertebrates, the pinealocytes have a sensory cilium developed to a different extent. The axonic processes of pinealocytes form ribbon-containing synapses on secondary pineal neurons, and/or neurohormonal terminals on the basal lamina of the surface of the pineal nervous tissue facing the perivascular spaces. Ribbon-containing axo-dendritic synapses were found in the rat, cat, guinea pig, ferret, and hedgehog. In the cat, we found GABA-immunoreactive interneurons, while the secondary nerve cells, whose axons enter the habenular commissure, were GABA-immunonegative. GABA-immunogold-labeled axons run between pinealocytes and form axo-dendritic synapses on intrapineal neurons. There is a similarity between the light and electron microscopic localization of Ca ions in the mammalian and submammalian pineal organs and retina of various vertebrates. Calcium pyroantimonate deposits--showing the presence of Ca ions--were found in the outer segments of the pineal and retinal photoreceptors of the frog. In the rat and human pineal organ, calcium accumulated on the plasmalemma of pinealocytes and intercellularly among pinealocytes. The formation of pineal concrements in mammals may be connected to the high need for Ca exchange of the pinealocytes for their supposed receptor and effector functions.


Neurobiological aspects of panic disorder.

The limbic system, temporal cortex and the locus coeruleus are important brain regions in the neuroanatomy of panic states. There is evidence for beta- and alpha 2-adrenoreceptor abnormalities and pre- and post-synaptic serotonergic alterations in panic disorder subjects. The anti-panic effects of chronic antidepressant drug treatment may relate to their down-regulation of various components of noradrenergic function and overall enhancement of serotonergic function. The efficacy of benzodiazepine agents in panic disorder and the altered sensitivity of panic patients to benzodiazepine agonists and inverse agonists suggest that alterations in benzodiazepine/GABA receptors may have a role in this disorder. A variety of other pharmacological agents also provoke panic, demonstrating that the biological origins of this disorder are quite diverse and complicated. Most importantly, a variety of new agents that selectively affect different components of these neurotransmitter/receptor systems are being developed. These novel agents offer future promise of greater efficacy with less adverse effects for individuals with panic disorder.


Beta-adrenergic agonists for acute, severe asthma.

To critically review the use of beta-adrenergic agonists in acute, severe asthma with particular focus on aerosol administration. English language articles published since 1971 on the use of beta-agonists for acute asthma. Studies were identified from bibliographies of book chapters, review articles, and other research articles. All studies (21 total) comparing systemic with inhaled beta-agonists were reviewed, regardless of their design or outcome. Selected studies highlighting specific aspects of beta-agonist use in acute asthma such as beta-agonists versus other bronchodilators, aerosol delivery, and intravenous beta-agonists were also reviewed. Performed subjectively by the authors with specific aspects of quality discussed within the body of the article. The beta-agonists provide superior bronchodilation in acute severe asthma compared with either the methylxanthines and/or anticholinergics. The majority of studies found aerosolized beta-agonists to be either as effective as or more effective than parenteral beta-agonists and to produce fewer adverse cardiovascular effects. Studies showing preference for parenteral therapy have either been of poor design or used low doses of an aerosolized beta-agonist. Based on studies of aerosol delivery, there is no advantage of jet nebulization over metered-dose inhalers; however, other aspects, including ease of administration, favor nebulization as the delivery method of choice. The articles recommending intravenous beta-agonists consist of a series of uncontrolled cases. Aerosolized selective beta 2-agonists are the bronchodilator treatments of choice for acute, severe asthma. Attention to the details of dosing and delivery are required for optimal results. The final dose and dosing interval are determined by the patient's response. Intravenous beta-agonists are hazardous and cannot be recommended.


Metered-dose inhalers and nebulizers in the acute setting.

This article evaluates the current literature comparing beta-adrenergic agonists administered via metered-dose inhalers (MDIs) with nebulizer devices in adult and pediatric patients. These studies focus on the acute treatment of asthma or chronic obstructive pulmonary disease in the emergency department and other acute care settings. English-language journal articles published between 1980 and 1991. Eight studies that compared beta-adrenergic agonists administered via an MDI or an MDI with a spacing device versus a nebulizer were identified. All of the studies were either poorly designed or had few subjects. By consensus of the authors, all were included in the review. Studies were assessed according to methodologic strength (e.g., prospective, comparative). Five studies found no differences between administration methods, one study found metered-dose inhalation to be superior, and another found nebulization to be superior based on observed improvements in pulmonary function tests. There were no significant differences in adverse-reaction rates. When surveyed, subjects preferred MDIs to nebulizers. There were marked variations in doses administered within and between studies. There was no consideration given to doses potentially delivered to the lungs. There is no significant difference between nebulizers and MDIs plus a spacer with regard to the administration of beta-agonists in the treatment of acute asthma. There are insufficient data to conclusively support the role of spacers in this setting. The choice of a specific delivery method at this time must be determined on an individual basis, taking into account the issues of cost, timeliness of administration, and personnel availability.


Application of restrained minimization, simulated annealing and molecular dynamics simulations for the conformational analysis of oligosaccharides.

The purpose of the present study was to determine the confidence with which the small number of 1H NMR nuclear Overhauser effect (NOE) distance constraints measurable across glycosidic linkages in oligosaccharides could be used for solution conformational analysis. This was assessed by use of these constraints in restrained molecular mechanical minimization of the tetrasaccharide Gal beta 1----4(Fuc alpha 1----3)Glc-NAc beta 1----3Gal, a model compound of the Lewis-X antigenic determinant. This presents a particularly severe test case in view of extreme resonance overlap and a dearth of inter-residue distance constraints. It is concluded that these constraints, when used in conventional restrained minimization, result in the generation of 'virtual conformations' and local minima about glycosidic linkages. However, these restraints are nevertheless found to be useful in the initial stages of a conformational analysis strategy involving restrained minimization combined with dynamical simulated annealing to define more accurately the global minimum energy configuration, together with molecular dynamics simulation to explore conformational mobility about this minimum. Theoretical ROE values calculated over the time course of the MD simulation, using a formalism appropriate for the time scale of the internal motion, are compared with those obtained experimentally in the oligosaccharide.


Structure and function of neuromuscular junctions in the vastus lateralis of man. A motor point biopsy study of two groups of patients.

The properties of neuromuscular junctions (NMJs) in the vastus lateralis of man have been studied in motor point biopsy samples and compared with those reported for lower vertebrates (frogs and mice). The patients studied had no convincing evidence of a primary disturbance of neuromuscular transmission or other neurogenic component. Morphological studies were made using a variety of methods at the light- and electron-microscope levels. The size of the presynaptic nerve terminal and the area of postsynaptic specialization were smaller, relative to the size of the muscle fibres, than in the lower vertebrates. In contrast, the extent of postsynaptic folding was greater. Intracellular recordings from single muscle fibres showed that the duration of synaptic currents was longer than in most other vertebrates so far studied and that the number of transmitter 'quanta' released by a single nerve impulse, about 20, was lower, probably reflecting the small size of the presynaptic terminals. The hypothesis is discussed that in man, a relatively weak effect of transmitter on the muscle fibre surface is amplified by voltage-dependent sodium channels which have been shown in the rat to be concentrated in the depths of the synaptic folds. The implications of this hypothesis for the interpretation of pathological findings in myasthenic syndromes are also discussed.


Adenosine modulation of dynorphin B release by hippocampal synaptosomes.

A rat hippocampal preparation enriched in mossy fiber synaptosomes was employed in an attempt to expose any relationship between endogenous adenosine and the release of dynorphin B-like immunoreactivity (DynB-LI). Presumptive blockade of purinergic receptors increased the spontaneous release of DynB-LI, and reducing synaptic adenosine by exogenous adenosine deaminase increased the K(+)-evoked release. Evoked release of DynB-LI was reduced by inhibitors of adenosine uptake and 5'-nucleotidase. Taken together, these data suggest that adenosine endogenous to hippocampal mossy fiber synaptosomes serves to inhibit the release of one of the peptide neuromodulators of this preparation, and provide support for the concept of autoregulation of release.


Role of the deep mesencephalic nucleus in the antinociception induced by stimulation of the anterior pretectal nucleus in rats.

The present study showed that the inhibitory effect on the tail-flick reflex (TF) of stimulating the deep mesencephalic nucleus (DpMe) was very similar to that produced by stimulation of the anterior pretectal nucleus (APtN). An electrolytic lesion of the ipsilateral DpMe greatly reduced the inhibitory effect of APtN stimulation on the TF. Furthermore, activating the neuronal cell bodies in DpMe but not the fibers of passage by microinjection of L-glutamate into this area was also shown to elicit inhibition of TF. On the other hand, inhibiting the neuronal cells in DpMe by microinjection of gamma-aminobutyric acid produced a marked reduction in the APtN-induced inhibition of TF, which was comparable to that produced by DpMe lesions. It is suggested that the APtN-induced antinociception is, at least in part, mediated via a relay through the DpMe.


[Decrease of pineal AMPc and TOH activity in the superior cervical ganglia after ablation of submaxillary glands in rats].

In order to investigate a possible functional relationship between the submandibular salivary gland (SSG) and the central nervous system (CNS), we have extirpated the salivary organs from thirty male rats. Twenty days after ablation both the pineal glands and the cervical superior ganglions (CSG) were dissected, homogenized and frozen until AMPc and TOH were assayed respectively. We observed a significant decrease in pineal AMPc (53.9 +/- 6.2 vs 76.1 +/- 7.6% of maximum value; p less than 0.02) which seems to be linked with a significant drop in TOH activity measured at CSG level (1.5 +/- 0.6 vs 3.7 +/- 0.9 nmoles of DOPA/h/pair GCS; p less than 0.03). Our results suggest that both findings might be due to the lack of NGF normally reaching the CSG from SSG. This data reinforces the idea of a functional link between SSG and CNS via the pineal gland.


Progression of Takayasu's aortitis in a young Japanese woman: serial angiographic study.

Progression of Takayasu's aortitis was observed by serial angiography over 2 years. Acute aortic regurgitation due to marked dilatation of the proximal ascending aorta was followed by aneurysmal dilatation of the distal ascending aorta. Aneurysmal dilatation of the carotid arteries then developed, with subsequent obstruction of both the bilateral vessels by thrombosis.


A cytoplasmic chaperonin that catalyzes beta-actin folding.

We have isolated a cytoplasmic chaperonin based on its ability to catalyze the folding of denatured beta-actin. The cytoplasmic chaperonin is organized as a multisubunit toroid and requires Mg2+ and ATP for activity. The folding reaction proceeds via the rapid ATP-independent formation of a binary complex, followed by a slower ATP-dependent release of the native product. Electron microscopic observations reveal a striking structural change that occurs upon addition of Mg2+ and ATP. The eukaryotic cytoplasm thus contains a chaperonin that is functionally analagous to its prokaryotic, mitochondrial, and chloroplastic counterparts.


Ocular beta-blockers in glaucoma management. Clinical pharmacological aspects.

Topical beta-blockers reduce the intraocular pressure (IOP) by blockade of sympathetic nerve endings in the ciliary epithelium causing a fall in aqueous humour production. Two types of topical beta-blockers are available for use in glaucoma: nonselective, which block both beta 1- and beta 2-adrenoceptors; and cardioselective, which block only beta 1-receptors. Of the beta-Blockers commercially available, timolol, levobunolol, metipranolol and carteolol are nonselective, and betaxolol is cardioselective. Twice-daily timolol is probably the most effective agent in lowering IOP, although levobunolol is equally effective and can be used once daily with little difference in effect. Carteolol is used twice daily and any theoretical advantage in diminished side effects conferred by its partial beta-agonist activity compared with timolol has not been fully substantiated. Metipranolol is effective twice daily and does not have partial beta-agonist activity. Betaxolol has an effect comparable to timolol in lowering IOP, but is less effective in some patients. beta-Blockers can be used with other antiglaucoma medications, but their combined action with epinephrine (adrenaline) is suspect, particularly in the case of the nonselective beta-blockers, and the effect should be assessed in patients on an individual basis. Local stinging can be a problem in some patients with betaxolol. The most serious side effects of beta-blockers are the exacerbation of chronic obstructive airways disease with nonselective agents and the precipitation of bronchospasm in some patients. Betaxolol seems relatively free of adverse respiratory effects, although this may be dose-related and extreme caution should still be exercised in patients with any history of respiratory illness. Because of the lower risk of precipitating side effects, betaxolol is probably the beta-blocker of first choice for use in glaucoma; timolol or levobunolol are reserved for patients who do not respond satisfactorily to betaxolol and are quite free of respiratory disease.


Inhibitory effect of taxol, a microtubule stabilizing agent, on induction of DNA synthesis is dependent upon cell lines and growth factors.

Growth-arrested rat fibroblasts, 3Y1, and human diploid fibroblasts, TIG-1, were induced to synthesize DNA by stimulation with various agents such as fetal bovine serum (FBS), epidermal growth factor (EGF), colcemid, or colchicine. Taxol, a microtubule-stabilizing agent, blocked the induction of DNA synthesis after stimulation with colcemid or colchicine in both cell lines. Taxol inhibited the induction of DNA synthesis after stimulation with FBS or EGF in TIG-1, but did not in 3Y1. 12-O-tetradecanoylphorbol-13-acetate (TPA) induced DNA synthesis in TIG-1, which was reduced only partly by taxol. Taxol stabilized or polymerized microtubules in both cell lines. These results indicate that the inhibitory effect of taxol on the induction of DNA synthesis varied among cell lines and among growth factors, and suggest that signal transduction processes may be differentiated by taxol sensitivity. In TIG-1 cells, when taxol was added within 6 h, about halfway into the initiation of DNA synthesis after the addition of FBS or EGF, the inhibition of DNA synthesis still occurred. Taxol did not inhibit the induction of c-fos and c-myc genes by FBS or EGF stimulation. Colchicine itself did not induce these genes in TIG-1. Thus, taxol appeared to inhibit the induction of DNA synthesis not by blockage in the early transduction process of the growth signal from the cell surface to nuclei but by blockage in processes operating in the mid- or late-prereplicative phase.


Study on stereospecificity of enzyme reaction related to peroxisomal bile acid synthesis in rat liver.

We examined stereoselectivities of enzymes related to bile acid formation in hepatic peroxisomes using two stereoisomers of 3 alpha,7 alpha,12 alpha-trihydroxy-5 beta-cholestanoic acid (THCA) and its coenzyme A (CoA) derivatives. The activity of acyl-CoA synthetase for 25 S-THCA was 1.4-times higher than that for 25 R-THCA. The difference was also observed after clofibrate-treatment. This activity was located in microsomes, differing from palmitoyl-CoA synthetase located in mitochondria, peroxisomes and microsomes. There was no stereoselectivity in the reaction of peroxisomal fatty acyl-CoA oxidase for THCA isomers, and the activity was one tenth of that for acyl-CoA synthetase. Considering the overall reaction of peroxisomal bile acid formation, the stereoselective difference observed in THCA-CoA synthesis should be denied. Thus, the previous finding that the overall formation of bile acid from THCA was not stereoselective was further confirmed. Furthermore, the activity for THCA oxidation was not induced by clofibric acid, suggesting that there would be different isozymes of peroxisomal acyl-CoA oxidase against THCA-CoA and palmitoyl-CoA.


Autoradiographic characterization of beta-adrenergic receptor subtype in the canine conduction system.

It has been hypothesized, based on physiological evidence, that there is a greater proportion of beta 2-adrenergic receptors on the myocytes of the conduction system when compared with the working myocardium. The purpose of these studies was to examine beta-adrenergic receptor subtype in the conduction system of the dog by using the technique of coverslip autoradiography. Scintillation studies of [125I]pindolol binding to ventricular sections demonstrated that binding was saturable (dissociation constant of 116 pM), had the correct order of potency for a beta-receptor, and was stereoselective. Both betaxolol (beta 1-selective) and ICI-118,551 (beta 2-selective) competition curves fit a two-site model in nonlinear curve-fitting analyses (78% beta 1-receptors). Autoradiographic studies determined that the myocytes of the sinoatrial node had approximately twice as many autoradiographic grains as the surrounding atrial myocytes. The myocytes of the atrioventricular bundle had a number of grains similar to the number in surrounding septal myocytes. Autoradiographic inhibition curves with betaxolol or ICI-118,551 demonstrated that both the sinoatrial node and the atrioventricular bundle had inhibition profiles similar to the surrounding myocytes (predominantly beta 1) but unlike the inhibition profiles of arterioles (predominantly beta 2). Calculations using the dissociation constants derived from the nonlinear curve-fitting analysis and the percent specific binding in the presence of 4 x 10(-7) M betaxolol or ICI-118,551 determined that the proportion of beta 1- to beta 2-receptors was the same (70-80% beta 1) when comparing the sinoatrial node and the surrounding atrial myocytes.(ABSTRACT TRUNCATED AT 250 WORDS)


Role of neutrophil adherence receptors (CD 18) in lung permeability following lower torso ischemia.

Ischemia and reperfusion of the lower torso lead to leukotriene- and neutrophil (PMN)-dependent lung injury characterized by lung PMN sequestration, increased permeability, and noncardiogenic edema. It is thought that PMNs require adhesion to endothelium to alter barrier function. This study tests the role of CD 18, the PMN adherence receptor, in mediating lung permeability after lower torso ischemia and reperfusion. Anesthetized rabbits (n = 9) underwent 3 hours of bilateral hind limb ischemia. Ten minutes after the release of the tourniquets, plasma leukotriene B4 levels increased to 395 +/- 85 pg/ml, higher than 129 +/- 35 pg/ml in controls (n = 9, p less than 0.01). At this time there was a reduction in circulating white blood cells (x 10(3)), 3.56 +/- 0.49/mm3 relative to 6.07 +/- 0.61/mm3 in controls (p less than 0.01). PMNs were sequestered in the hind limbs, indicated by increased myeloperoxidase activity of 1.06 +/- 0.19 units/g compared with 0.56 +/- 0.09 units/g in controls (p less than 0.05). Four hours after tourniquet release, PMNs were sequestered in the lungs, 52 +/- 4 PMNs per 10 high-power fields, a value higher than 31.5 +/- 3 PMNs per 10 high-power fields in controls; bronchoalveolar lavage fluid protein content increased to 554 +/- 90 micrograms/ml relative to 277 +/- 46 micrograms/ml in controls; and there was lung edema, measured by increased wet weight-to-dry weight ratios of 5.19 +/- 0.10, higher than 4.29 +/- 0.21 in controls (all p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)


Postnatal changes in AMPA receptor regulation by phospholipase A2 treatment of synaptic membranes: temporally differential effects on agonist and antagonist binding.

Previous results have indicated that phospholipase A2 (PLA2) treatment of telencephalic membranes produced opposite effects on [3H]amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) binding in neonatal and adult rats. In the present study, we compared the effects of PLA2 treatment of telencephalic membranes on the binding characteristics of agonists and antagonists of the AMPA receptors in the developing rat brain. Whereas PLA2 treatment of telencephalic membranes from postnatal day (PND) 5 and 10 animals produced an important decrease in [3H]AMPA binding, the same treatment performed on PND 20, 25 and adult membranes resulted in a marked increase in [3H]AMPA binding; the shift from decreased to increased [3H]AMPA binding occurred at about PND 15. In contrast to [3H]AMPA binding, [3H]6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) binding was substantially reduced following PLA2 treatment at PND 5, 10 and 20, and effect due to a decrease in the maximal number of [3H]CNQX binding sites. In adult membranes, the effect of PLA2 treatment on [3H]CNQX binding was markedly reduced when compared to neonatal membranes. Pretreatment of synaptic membranes with PCMBS (a sulfhydryl reagent) increased [3H]AMPA binding in both young (PND 10) and adult telencephalic membranes, without significantly changing [3H]CNQX binding. The various effects of PLA2 treatment on agonist and antagonist binding did not appear to be due to major differences in the pharmacological properties of the AMPA receptors at different ages. The present results indicate that the characteristics of the binding sites for agonists and antagonists of the AMPA receptors are differentially modulated by the lipid environment during the postnatal period.(ABSTRACT TRUNCATED AT 250 WORDS)


Ontogenesis of pyroglutamyl peptidase II activity in rat brain, adenohypophysis and pancreas.

Pyroglutamyl peptidase II (PPII; E.C. 3.4.19.-) is a highly specific membrane-bound ectoenzyme degrading thyrotropin releasing hormone (TRH). The ontogenesis of this enzyme was measured in rat brain regions, adenohypophysis and pancreas. In hypothalamus PPII activity was maximal at day 8 postnatal, decreasing to adult values at day 45. The postnatal ontogenic patterns in posterior cerebral cortex and hypothalamus were similar. In olfactory bulb, two peaks of activity were observed (3th and 22nd day) while in adenohypophysis it appeared only at day 8, increased to day 30, decreasing thereafter to adult values.


Increased tyrosine hydroxylase immunoreactivity in the rat cortex following prenatal cocaine exposure.

Cocaine has been found to be a neurobehavioral teratogen in both animals and humans. In this study the effects of cocaine on the developing catecholamine systems were examined. Rats were treated gestationally with cocaine (40 mg/kg s.c.) or saline from gestational day 13 until parturition. On postnatal day 28, tyrosine hydroxylase immunocytochemistry was performed. Increases in catecholamine fiber densities were observed in the hippocampus, anterior cingulate cortex, and parietal cortex in cocaine-treated animals. These findings may explain some of the behavioral alterations seen following prenatal cocaine exposure.


Polymorphisms at the GLUT2 (beta-cell/liver) glucose transporter gene and non-insulin-dependent diabetes mellitus (NIDDM): analysis in affected pedigree members.

The precise genetic defects underlying the etiology of non-insulin-dependent diabetes mellitus (NIDDM) have yet to be identified. The beta-cell/liver glucose transporter gene GLUT2 represents a good candidate for the etiology of the disease, being involved in the glucose signalling for beta-cell insulin release. Population association studies of the GLUT2 gene in NIDDM have so far yielded controversial results. In order to determine the possible contribution of this gene to the inheritance of NIDDM, we have employed a new approach, where two polymorphic markers of the GLUT2 locus, detected with the restriction enzyme Taq-1, were examined for linkage with the disease in a group of 22 Italian pedigrees with affected members (n = 50). Departure from independent segregation between markers and disease was analyzed by the Affected-Pedigree-Members (APM) statistical method. Furthermore, association analysis between the Taq-1 RFLPs at the GLUT2 locus and NIDDM was performed in a group of diabetics with a strong family history, comprising the 22 probands and 23 other diabetics with an affected first-degree relative. The results indicate that there was no segregation distortion between the Taq-1 markers of the GLUT2 gene and the disease in the pedigrees examined. Also, no significant difference in genotype distribution, haplotype and allele frequencies was found between diabetics and controls for the two Taq-1 RFLPs. We conclude that genetic variation at the GLUT2 transporter gene is unlikely to contribute in a major way to the inheritance for NIDDM in this Italian population.


Deletions, duplications and novel restriction fragment length polymorphism in Duchenne and Becker muscular dystrophies.

To determine the mutations of Southern Chinese with Duchenne and Becker muscular dystrophies (DMD, BMD), we analysed 28 DMD and BMD patients in 24 unrelated families for intragenic deletions and duplications by using cDNA probes covering the entire 14 kb of the dystrophin gene. Deletions were detected in nine unrelated patients (seven patients by probe 8 and two by probe 2b-3). Gene duplications were detected by probe 1-2a in two patients with the duplication bands confirmed in both Hind III and Bgl II digests and by densitometry. A third patient was found to have a junction fragment with Bgl II and a duplication band with Hind III by probe 5b-7. Therefore 50% of the 24 unrelated families were found to have either deletions or duplications. A previously undescribed restriction fragment length polymorphism (RFLP) was found in one family with probe 5b-7 in Bgl II digests which was found to segregate with the disease phenotype. This new RFLP was not detected in over 70 unrelated X chromosomes we have examined so far, and appeared to be "private" for this family. The presence of this new restriction site may or may not be the mutation responsible for the disease phenotype.


Impaired proliferative capacity and abnormal cytokine profile of naive and memory CD4 T cells from HIV-seropositive patients.

Purified naive and memory CD4 T cells from healthy donors, HIV+ asymptomatic carriers and AIDS patients were examined for their proliferative activity and their pattern of cytokine secretion (IL-4, IL-6, interferon-gamma (IFN-gamma) and tumour necrosis factor-alpha (TNF-alpha)) upon stimulation with phytohaemagglutinin (PHA), phorbol myristate acetate (PMA) and cross-linked anti-CD3 MoAb, in the presence of recombinant IL-2 (rIL-2). We found a decrease in the proliferative capacity of naive CD4 T cells following stimulation with PHA and PMA, and a sharp decline in this response upon cross-linked anti-CD3 stimulation in both subsets, although it predominated in the naive subpopulation. In AIDS patients, less pronounced impairment of thymidine uptake by the naive subset was found upon PHA and cross-linked anti-CD3 MoAb stimulation. In addition, an altered secretion pattern of the different cytokines was observed, consisting of abnormal secretion of IL-6 by both naive and memory cells, an abnormal pattern of IFN-gamma secretion and frequent loss of detectable IL-4 production by HIV patients. These abnormalities were even more pronounced in AIDS patients than in the asymptomatic carriers. Overall, our results extend previous reports indicating functional impairment of memory CD4 subsets in HIV+ subjects by showing that this impairment involves naive CD4 T cells.


Tumour necrosis factor-dependent parasite-killing effects during paroxysms in non-immune Plasmodium vivax malaria patients.

Plasmodium vivax malaria infections in non-immune individuals manifest as periodic clinical episodes of fever with chills and rigors known as paroxysms. We have demonstrated that in non-immune patients the period of paroxysm is associated with the transient presence of plasma factors which kill gametocytes, the intra-erythrocytic sexual stages of the malaria parasite which transmit the infection from humans to mosquito, rendering them non-infectious to mosquitoes. Gametocyte killing in paroxysm plasma is mediated by tumour necrosis factor (TNF) acting in conjunction with other essential serum factor(s). Plasma TNF levels were elevated during a paroxysm. In semi-immune individuals from a P. vivax-endemic area clinical symptoms of malaria are mild and the parasite killing factors are not induced during paroxysm. Serum TNF levels were correspondingly lower in endemic patients during a paroxysm. Human peripheral blood mononuclear cells (PBMC) can be stimulated in vitro by extracts of P. vivax blood stage parasites to produce TNF and associated parasite killing factor(s), thus simulating in vitro the events that occur during a paroxysm, this being the release of parasite exo-antigens by rupturing schizonts and the subsequent induction of PBMC to produce TNF and other parasite-killing factors. We were able to show that convalescent serum from P. vivax semi-immune individuals block the induction of TNF and parasite-killing factors by malaria antigens in vitro, presumably through antibodies that neutralize parasite exo-antigens. Thus, individuals living in malaria-endemic areas appear to acquire clinical immunity to malaria by avoiding their induction during infection; we have shown that one such mechanism is the neutralization of parasite exo-antigens that induce the production of parasite killing factors.


Antineutrophil cytoplasmic antibody (cANCA and pANCA) titers in relation to disease activity in patients with necrotizing vasculitis: a longitudinal study.

The significance of the antineutrophil cytoplasmic antibodies, cANCA or pANCA, in relation to disease activity in various types of necrotizing vasculitis was assessed in a longitudinal study. Twenty patients, 14 cANCA positive and 6 pANCA positive were followed for up to seven years. Eleven of the 14 cANCA positive and two out of the 6 pANCA positive patients had Wegener's granulomatosis, two other cANCA patient and one pANCA patient had systemic vasculitis, whereas one cANCA and three pANCA positive patients had disease limited to the kidneys. Disease activity was expressed as a numerical index DAI, based on extent and type of organ involvement (DAI greater than or equal to 5 = active disease). At the time of disease debut the DAI ranged between 11 and 32, cANCA titers between 18 and greater than 1000 U/l, and pANCA titers between 51 and 630 U/l in individual patients. With clinical recovery both c- and pANCA titers declined. Clinical remission (DAI less than 5) occurred within a mean of 2.5 months (range 1 to 4). Almost simultaneously cANCA titers turned negative (less than 10 U/l), although one patient remained positive for seven years. pANCA titers remained positive for long periods of time in 5/6 patients. Five cANCA patients had clinical exacerbations, which in two patients were preceded by seroconversions from - to +, lasting 3 and 12 months before the clinical relapse. One additional cANCA patient had two serological relapses without signs of clinical activity during a 2-year follow-up. In conclusion, we saw a close correlation between DAI and cANCA titers in individual patients: with decreasing clinical activity (DAI), the cANCA titers decrease. Interindividual variation is great; despite a high DAI, the cANCA titers may be relatively low. Serological relapses may indicate forthcoming clinical relapses. Relapses may develop as late as 5-6 years after onset. pANCA titers, on the other hand, tend to remain positive for long periods, despite clinical remission.


CD4+ T cells clear virus but augment disease in mice infected with respiratory syncytial virus. Comparison with the effects of CD8+ T cells.

Respiratory syncytial (RS) virus-specific T cell lines were derived from the spleens of BALB/c mice primed by intranasal infection with RS virus. The lines were expanded by repeated antigenic stimulation in vitro, and separated into CD4+ and CD8+ T cell-enriched fractions by immunomagnetic adhesion. The effects of passive transfer of these fractions into RS virus infected mice were observed. The most severe immunopathological changes were seen in mice receiving CD4+ cells. Transfer of CD4+, CD8+ or both cell fractions caused RS virus-infected mice to become ill and lose weight. Both cell lines caused an increase in the severity of lung pathology (as monitored by bronchoalveolar lavage) with the appearance of lung haemorrhage and polymorphonuclear cell efflux. In addition, recipients of CD4+ cells developed striking pulmonary eosinophilia. In CD4+ cell recipients, 5 x 10(5) cells were sufficient to decrease lung virus titre, whereas 2 x 10(6) CD8+ cells were needed to produce a similar effect. The unseparated T cell line and the CD4+ cell fraction secreted significant amounts of IL-3, IL-4 and IL-5 (P less than 0.001). High levels of IL-2 were produced only by the unseparated T cell line. The CD8+ cell fraction secreted IL-3 only. The results show that, cell-for-cell, CD4+ cells are more anti-viral and more immunopathogenic than CD8+ cells in RS virus infected mice. Such effects may have contributed to the augmented disease seen in some infants vaccinated against RS virus.


Case report: therapeutic bronchial artery embolization in a case of Takayasu's arteritis.

The pulmonary arteries are frequently involved in Takayasu's arteritis with a reported incidence of 41-100% in affected individuals. Patients are usually asymptomatic, however, often despite extensive pulmonary involvement. We describe a patient with Takayasu's arteritis who presented with haemoptysis caused by bronchial artery hypertrophy secondary to occlusive pulmonary arterial disease, who was treated by bronchial artery embolization. This is, to our knowledge, the first reported case of haemoptysis secondary to Takayasu's arteritis successfully treated by embolization.


The pivotal role of the plasma membrane-cytoskeletal complex and of epithelium formation in differentiation in animals.

1. If a few exceptions are disregarded, the several somatic cell types of a differentiated organism all have an identical genome. They all differ in their plasma membrane-cytoskeletal complex. 2. Differences in plasma membrane properties usually result in differences in ionic concentrations/activities in the cytoplasm and nucleoplasm. A basic question therefore is whether there exists a causal relationship between the ionic environment of the nucleus and differential gene expression/protein synthesis. 3. Development is switched on by a "Ca2+ explosion", often accompanied by pH changes and plasma membrane depolarisation. The penetration of the spermatozoon in the plasma membrane acts as a trigger. 4. All animal species develop from a blastula. At this stage they organise themselves as an epithelium enclosing an inner (fluid) compartment. This suggests that epithelium formation is absolutely essential in animal development. 5. As development proceeds, more and more compartments, lined by different epithelia, are formed. Differentiated organisms largely consist of folded epithelia. Some cells leave their original epithelial environment and become free floating (e.g. blood cells) or engage in other types of organisation. 6. Epithelial cells have the ability to segregate some membrane proteins, e.g. receptors, ion pumps, ion channels etc., so as to make selective transcellular transport possible. The cytoskeleton plays an important role in this segregation and in the interconnection of epithelial cells. 7. Transembryonic electric currents which have been measured by the vibrating probe technique, are due to electrogenic ion transport by epithelia. 8. Segregation of membrane proteins is not an exclusive property of epithelial cells but it is probably a property of all animal cell types, single cells inclusive; asymmetry is the rule, symmetry--if it exists at all--the exception. 9. Differences in several plasma membrane proteins (receptors, ion transporting molecules, cell adhesion molecules and signal transducing systems) are not only causally related to differential transcellular transport but also indirectly to differential protein synthesis and hence to differentiation. There are already a few well documented examples of "electrical" control of gene expression. 10. The major "strategy" which applies in differentiation seems to be to keep the genome constant but to change over and over its ionic and macromolecular environment, both acting in a complementary way. The first one may be considered as the coarse tuning mechanism of gene expression-protein synthesis, the second as the fine one. In our opinion this might be a principle universal to differentiation processes in all animal species.


The effects of surfactants on the permeability of isolated perfused fish gills to urea.

1. The diffusional transfer capacity of [14C]urea in isolated perfused trout (Oncorhynchus mykiss) gills in the presence of sodium n-dodecylsulphate (SDS), n-dodecyltrimethylammonium bromide (DTAB) and p-t-octylbenzene oxyethylene10 (Triton X-100) has been measured over a range of surfactant concentrations. 2. Urea has been shown to be transported transcellularly through the respiratory cells of the gill secondary lamellae by passive diffusion. Each surfactant was found to markedly increase the rate of diffusion and the diffusional transfer capacity reached a steady-state at a particular surfactant concentration. 3. The steady state flux was increased by surfactant in the sequence DTAB greater than SDS greater than Triton X-100 and the surfactant concentrations in terms of the critical micelle concentration (CMC) at which the diffusional transfer capacities reached limiting values were 0.92 x CMC (SDS), 0.53 x CMC (DTAB) and 2.5 x CMC (Triton X-100). 4. Compared to interactions between isolated epithelial cells and the surfactants, the rates at which the surfactants changed the urea flux were slow, suggesting that the mucus layer plays a significant role in protecting the epithelial cells of the secondary lamellae from disruption. 5. Relative to the other surfactants, DTAB had the most marked effect on both the rate of flux change and on the magnitude of the change, at concentrations which are low relative to the CMC, suggesting a more specific interaction with the negatively charged mucus layer consistent with the toxic effects of quaternary ammonium compounds on aquatic organisms.


Effects of somatostatin on inositol-1,4,5-trisphosphate content in mouse spleen lymphocytes.

1. The effects of somatostatin-14 (SS) and diazepam in vitro on the content of inositol-1,4,5-trisphosphate (IP3) in mouse spleen lymphocytes were investigated. 2. It was found that the exposure of mouse spleen lymphocytes in vitro to SS sharply diminished their IP3 level. 3. Diazepam had no effect on lymphocytes IP3 content. 4. The inhibition of phosphatydyloinositol (PI) breakdown was suggested as one of the mechanisms of the physiological SS action.


Interaction between cortisol and cortisol-binding protein in silver foxes (Vulpes fulvus).

1. Selection of silver foxes for domestic behaviour resulted in the parallel lowering of both cortisol and cortisol-binding protein (CBP) levels in the blood plasma. 2. During seasonal cycles (summer-winter) and after stress an increase in cortisol levels is followed by a decrease in CBP activity. 3. It is concluded that there are two types of interaction between cortisol and CBP in silver foxes: parallel changes in the process of domestication and opposite changes under the influence of environmental factors.


Effect of antiserum to rat adipocytes on growth and body composition of the rat.

1. Antibody against intact adipocytes was produced in sheep and was capable of binding to intact rat adipocytes using an immunofluorescent assay. An antibody dose level was established and subsequently used in vivo. 2. This study demonstrated inconsistent response of passive immunization procedures with crude antibody against rat adipocytes. Establishment of specific protocols for each antibody preparation would be essential for using this immunological approach in body fat reduction.


Antigenic cross-reactions among immunoglobulin of diverse vertebrates (elasmobranchs to man) detected using xenoantisera.

1. Antisera raised in rabbits and goats against intact immunoglobulins or their constituent light and heavy chains from man, mouse and the galapagos shark (Carcharhinus galapagenesis) were tested for their reactivity with immunoglobulins of elasmobranchs, other lower vertebrates and eutherian and prototherian mammals. 2. Xenoantisera directed against human heavy chain isotypes allowed the serological identification of IgM and IgG immunoglobulins in the echidna (Tachyglossus aculeatus), a monotreme which is one of the most primitive species of extant mammals. 3. The antisera to heavy chains reacted to varying degrees with purified immunoglobulins of non-mammalian species, including the chicken, teleost fish and elasmobranchs in a fashion that was specific for immunoglobulins, but was not related to defined human isotypic markers. 4. The reactions of some antisera seem to skip species known to possess homologous immunoglobulins. 5. Antisera directed against isotypic markers of human kappa and lambda light chains reacted with shark light chains in a manner that was specific for light chain determinants but was not isotype-related. 6. Antisera directed against heavy chains of either sharks or mammals reacted with heavy chains, but not with light chains of diverse species. 7. A rabbit antiserum specific for shark light chain reacted with human and murine monoclonal lambda chains and with two synthetic peptides corresponding to human V lambda Fr3 and Fr4 sequences. 8. These results establish that a variety of antigenic markers including conformational and linear determinants can be shared among immunoglobulins of vertebrates species that had an ancestral divergence more than 400 million years ago.


Stimulation of hepatic thyroxine 5'-deiodinase activity in rainbow trout (Oncorhynchus mykiss) by Pacific salmon growth hormone.

1. Growth hormone extracted from Pacific salmon pituitaries (sGH) was injected intra-peritoneally into rainbow trout to determine sGH effects on plasma levels of thyroxine (T4), 3,5,3'-triiodothyronine (T3) and properties of the hepatic 5'-deiodinase enzyme (5'-D) responsible for T4 to T3 conversion. 2. After 24 hr, sGH (0.1 or 0.5 microgram/g) did not alter the plasma T4 level or 5'-D, Km, but elevated plasma T3 and especially 5'-D Vmax. 3. Thus sGH, like the previously tested human GH, acutely enhances the potential for extra-thyroidal T3 production in trout by increasing the functional level of hepatic 5'-D, but without changing the plasma T4 level.


Heat exchange by the pinna of the African elephant (Loxodonta africana).

1. Surface temperatures of the pinnae of four female African elephants were measured at ambient temperatures between 14 and 32 degrees C using infrared thermography. Instantaneous heat losses calculated using those values ranged from 10.67 to 76.2 W under the observed conditions. 2. Using a value of 17 kcal/kg/day, those heat losses account for 0.65-4.64% of the animals' standard metabolic rates, considering one side of one ear only. 3. A model of heat flow across a flat vertical plate was constructed and compared to the actual values. Up to 100% of an African elephant's heat loss needs can be met by movement of its pinnae and by vasodilation. 4. Thermography indicates that the temperature distribution pattern across the pinna changes with ambient temperature and that areas of specialized motor control exist.


Elements of acid-base balance in the European bison, Bison bonasus (L.) in the winter period.

1. In the blood of 65 European bison divided into four groups (Group 1, 0-3-year-old males; Group 2, 0-3-year-old females; Group 3, mature bulls, over 3 years old; Group 4, mature cows, over 3 years old) the values of pH, pCO2, pO2, base excess and bicarbonate level were studied. 2. The studies were carried out in winter months, from mid-December to mid-April. 3. No significant differences in parameters studied were found between young males and females. 4. All indices found in mature bulls and cows showed significant differences between all other groups. 5. Most of parameters examined here in European bison are comparable with those found in other big ruminants.


Metabolic rate and evaporative water loss at different ambient temperatures in two species of fox: the red fox (Vulpes vulpes) and the Arctic fox (Alopex lagopus).

1. Resting metabolic rate (RMR) and evaporative water loss (EWL) of adult red and arctic foxes were determined over ambient temperature (Ta) ranges of -13-37 degrees C and -5-30 degrees C as oxygen consumption and amount of water in expired air using an open flow system. 2. The average RMR was 2.60 +/- 0.14 W/kg for the winter red fox, 2.59 +/- 0.14 W/kg for the summer red fox, and 2.35 +/- 0.11 W/kg for the winter arctic fox. 3. The rate of increase of RMR was significant (P less than 0.05) only for Ta range above 27 degrees C. The slopes for this Ta range were 0.152 for the winter red fox, and 0.283 for the winter arctic fox. 4. The upper critical temperature (Tuc) of the red fox is probably between 30 and 32 degrees C. The Tuc of the arctic fox is probably between 26 and 28 degrees C. The lower critical temperatures (Tlc) were not reached. 5. A strong linear relationship between the EWL and Ta was found for Ta range above 27 degrees C. The slopes for this Ta range were 0.523 for the winter red fox, and 1.025 for the winter arctic fox. 6. Probably, there are neither significant intraspecific seasonal nor interspecific differences in the RMR and EWL. The two species seem to differ only in their critical temperatures.


Effects of arginine vasotocin, cortisol and adrenergic factors on water balance in the toad Bufo bufo: physiology or pharmacology?

1. The threshold level of exogenous arginine vasotocin (AVT) for increasing the cutaneous water permeability in Bufo bufo was several times higher than has been measured in dehydrated frogs and toads. 2. The beta-agonist isoproterenol was without effect on water balance in hydrated toads but increased the response to large doses of AVT by 25-50%. 3. Implantation of cortisol pellets in water-acclimated toads enhanced the cutaneous water permeability by a factor of 2-3. 4. It is concluded that understanding of water balance in terrestrial anurans may depend upon the unravelling of mechanisms that integrate cutaneous water transport and diuresis.


Hypoxia-induced changes in electrophysiological responses and associated calcium movements of flounder (Platichthys flesus) heart and gut.

1. Flounder hearts subjected to hypoxic conditions showed a reduction in contractile force. After 85 min in hypoxia-inducing salines, hearts were almost totally quiescent. 2. Membrane potentials were seen to change in hearts during hypoxic conditions. The action potential gradually declined in height but increased in duration and the resting membrane potential was also seen to decrease. 3. Sections of flounder gut, when subjected to identical hypoxic conditions as the hearts, showed no significant changes in either mechanical activity or membrane potential. 4. Heart and gut tissues were placed in hypoxia-inducing salines containing 45Ca either immediately or after a previous 3 hr exposure to hypoxia-inducing salines. Uptake of 45Ca increased beyond control values in both heart and gut subjected immediately to hypoxia. After 3 hr hypoxia exposure, uptake of 45Ca was reduced in heart but was not significantly different from controls in gut. 5. The changes seen in 45Ca uptake were far greater in heart tissue than in gut tissue. 6. In all physiological parameters examined, the gut tissue of the flounder is far more tolerant to hypoxic conditions than heart tissue.


The effects of hypoxic stress on the fine structure of the flounder heart (Platichthys flesus).

1. Flounder hearts were examined by conventional transmission electron microscopy. Hearts showed clear evidence of a coronary circulation but no intrinsic conduction network or innervation was detected. 2. The box-like cells showed many surface inpocketings and many scattered glycogen granules and membrane bound liposomes. The nucleus position was variable and the cells contained numerous small ovoid mitochondria. 3. Dark staining intercalated discs with clear nexal and desmosomal areas separated individual cells. 4. No organized T-tubular system or sarcoplasmic reticulum was present but the cells displayed abundant surface vesicles which may perform the physiological role of the latter. 5. Hearts of flounder subjected to 3 weeks hypoxia showed striking changes to the mitochondria which were smaller than in controls and there was evidence of increased mitochondrial budding and also evidence of mitochondrial necrosis. 6. Hypoxically-stressed hearts exhibited normal liposome populations but increased glycogen granule deposits. These hearts also showed evidence of myofibril degeneration with torn Z discs and with a build up of fibrous material. 7. It is concluded that the cellular damage seen in hypoxic cells may be due to excessive calcium accumulation or increased catecholamine release.


Influence of blood sample collection on the haematocrit value of two teleosts: rainbow trout (Oncorhynchus mykiss) and European sea bass (Dicentrarchus labrax L.).

1. The effects of the volume of blood extracted, number of extractions, and weight upon the haematocrit value of a freshwater teleost (Oncorhynchus mykiss) and a marine one (Dicentrarchus labrax) have been studied. 2. For trout with the same weight, the increase in the volume of blood extracted results in a significant increase in the haematocrit value. 3. For the same volume of blood extracted--in trout of the same weight--the second extraction results in a significant increase in the haematocrit value. 4. In sea bass, haematocrit variations in relation to the aforementioned parameters are not produced. 5. For both species there is a positive and significant correlation between weight (W) and haematocrit value (Ht), which coincides with the Ht = aWb model, in the range of weights used in this study.


Skeletal muscle capillarization and fiber types in urban and homing pigeons (Columba livia).

1. Fiber types, capillary supply and other morphometrical parameters were analysed in pectoral, gastrocnemius and pronator muscles of homing and urban pigeons. 2. The two kinds of birds were analysed before and after a restrainment period of at least 5 months. 3. Only slight differences in fiber type frequencies were noted between urban and homing pigeons in control conditions. 4. The effect of restrainment on the different parameters studied was unclear in gastrocnemius and pronator muscles and negligible in M. pectoralis. 5. Mean diffusion distances for oxygen from capillaries were smaller in oxidative fibers; also, vascularization indexes were higher for these fiber types. 6. The contribution of each fiber type to total sectional ara of muscle remains stable in spite of fiber type frequencies heterogeneity.


Characterization of the transport of tri- and dicarboxylates by pig intestinal brush-border membrane vesicles.

1. Transport of citrate and fumarate across the pig intestinal brush-border membrane (BBM) was investigated using isolated BBM vesicles. 2. Citrate and fumarate uptake was stimulated by an inwardly directed Na+ gradient consistent with Na+/citrate (fumarate) co-transport. Cis-inhibition and trans-stimulation experiments strongly suggest the existence of a common transport site for tri- and dicarboxylates. 3. The protonated forms of citrate (citrate-1, citrate-2) seem to be much better transported than citrate-3, indicated by the strong stimulation of citrate uptake at an extravesicular pH of 5.6 compared to pH 7.8. 4. Uptake of tri- and dicarboxylates seems to be potential-sensitive since citrate and in particular fumarate transport was enhanced by an inside negative potential difference. 5. Kinetics of succinate transport revealed a single carrier-mediated component with apparent kinetic constants of 0.43 nmol/mg protein-3 s (Vmax) and 0.14 mmol/l (Km).


Effects of hypoxia on renal function in carp, Cyprinus carpio.

1. Changes in urine and blood properties and heart rate (HR) of carp were investigated during and following hypoxia. 2. When carp were exposed to hypoxic conditions, urine flow in some carp increased immediately. However, it decreased gradually with time. Osmotic pressure and Na+, Cl-, Ca2+ and Mg2+ levels in urine increased in contrast to urine flow. K+, P, ammonium, and lactic acid levels in urine increased gradually. 3. When carp lost their balance, blood pH and plasma K+ were lower, and RBC, Ht, Hb, Mg2+, P, ammonium, lactic acid, and glucose in plasma were higher than those of the control. 4. As water-dissolved oxygen level was restored, urine flow increased immediately and soon decreased to the control rate. Other urine properties showed higher values than the initial levels and decreased with time. No significant change was found in urinary protein. 5. The relationship between HR and urine flow is discussed.


Distribution of 125I-endothelin-1,-2,-3 binding sites in mammalian kidneys.

1. Binding sites for 125I-labeled endothelin (ET) isopeptides ET-1, ET-2 and ET-3 were visualized by autoradiography in the kidneys of man, baboon, rhesus monkey, tree shrew, pig and rat. 2. Highest levels of binding for the three isoforms were observed in the glomeruli, cortical and medullary vessels of baboon, rhesus monkey, pig and rat, whereas there was no noticeable labeling of glomeruli in human and tree shrew kidneys. 3. The enrichment of binding sites depended on the species and peptide investigated, suggesting different affinities and/or densities of a heterogenous population of renal ET receptors.


A comparison of methods of measuring chromatic activity in the integument of a teleost.

1. Pallor induced by noradrenalin (5 x 10(-7) mol/kg) injection of dark Pseudopleuronectes americanus was measured as increase in skin reflectance and the degree of melanosome aggregation determined. 2. Mean skin reflectance increased approximately 30% on the Munsell neutral value scale. 3. At maximal mean reflectance, melanosomes were fully aggregated in most epidermal melanophores, but displayed more variable aggregation in dermal melanophores. 4. Individual reflectance estimates did not provide consistent indications of melanosome aggregation at maximum pallor. 5. The relative merits of reflectance and cellular methods of estimating melanophore activity are discussed with particular reference to this species.


Hematologic and blood chemistry data for the prairie dog (Cynomys ludovicianus).

1. The prairie dog has been used extensively for the study of gallstone genesis and gallstone dissolution therapies, and has recently been implicated in an effort to prevent total parenteral nutrition-associated cholelithiasis with intravenous chenodeoxycholate. 2. Towards this effort, it is important that a range of normal blood chemistry values be reported for the prairie dog. This paper reports the mean values for a complete blood cell count, electrolytes, blood urea nitrogen, creatinine, calcium, phosphorus, liver enzymes, total bilirubin, protein, albumin, cholesterol, triglycerides and lipids for 45 adult prairie dogs. 3. The prairie dog has normochromic, microcytic blood with an increased number of red blood cells. The prairie dog also has a high concentration of small platelets. 4. The prairie dog has a higher CO2 concentration with a slightly increased potassium concentration than is found in man. The anion gap is 12 with a calculated serum osmolality of 316. The BUN concentration is elevated with a 3-fold increase in the AST concentration. 5. The prairie dog has lower serum values for cholesterol, VLDL and LDL cholesterol than man. In the prairie dog, HDL cholesterol consists of 67% of the total cholesterol concentration and the LDL and HDL ratio is 0.3.


The vasopressor action of angiotensin in the snake Bothrops jararaca.

1. Carotid blood pressure from anesthetized B. jararaca snakes was recorded in order to study angiotensin action in this reptile. 2. Whereas [Asn1,Val5] AII and AIII were less potent than [Asp1,Ile5] AII and [Asp1,Val5] AII, [Sar1,Ile5] AII was slightly more potent. 3. Captopril abolished the responses to AI (0.01-3 micrograms/kg). 4. [Sar1,Ala8] AII was uneffective but [Sar1,Leu8] AII or phenoxybenzamine were able to reduce AII vasopressor responses. 5. These results led to the conclusion that the vasopressor response of AII in B. jararaca is due to an interaction with its own receptor but, part of the AII receptor population seems to be coupled to the sympatho-adrenal system. Moreover, structural requirements seem to be necessary for the AII response in B. jararaca.


Effects of salt acclimation on water and urea permeabilities across the frog bladder: relationship with intramembrane particle aggregates.

1. In salt-acclimated frogs, water and urea bladder permeabilities are markedly higher than in tap water-acclimated animals. 2. Intra-membrane particle aggregates (IMPA) cover an unusually large surface area of the salt-acclimated frog bladder apical plasma membrane. 3. In saline-adapted animals, proteins extracted from the apical plasma membrane contain additional species of 19, 26, 31 and 53-61 kDa. These proteins might be related to the water channels contained by IMPA.


Relationships between calbindin (Mr 28,000) and calcium transport by the eggshell gland.

1. Eggshell density (mg/cm2) and eggshell gland calbindin decreased in the aged hens. 2. Aged hens which laid eggs with a low shell weight and shell density had significantly lower intestinal and eggshell gland calbindin as compared with those which laid eggs with a high shell weight and shell density. 3. Significant correlations were found in aged hens between duodenal or eggshell calbindin and shell weight or shell density. 4. The results suggest that: (a) aged hens forming light shells absorbed calcium with a lower efficiency than those forming heavy shells or than young hens; (b) the decline in shell density in the aged hens is caused by a physiological calcium deficiency or by a defect in the hens' ability to alter calbindin synthesis in response to calcium needs; (c) in the aged hens, the deposition of calcium into the eggshell is dependent on, or at least associated with, eggshell gland calbindin.


Effects of injections of L-alanine, L-glucose and L-ascorbic acid in newly-hatched turkey poults on glucose metabolism.

1. Effects of subcutaneous injections of either L-glucose, L-alanine or L-ascorbate into newly-hatched, fasted turkey poults were examined. 2. There were no effects of injections on blood glucose concentrations at 20 hr post-injection. 3. Glucose injections inhibited hepatic glucose-6-phosphatase activity while alanine injections did not. 4. Both glucose and alanine injections enhanced hepatic glycogen reserves at 20 hr post-injection.


The effects of dietary deprivation on body temperature and oxygen consumption in black-tailed prairie dogs (Cynomys ludovicianus).

1. Body temperature (Tb) and oxygen consumption (VO2) were compared between fed (Control), food and water deprived (FWD), and water deprived (WD) black-tailed prairie dogs, in the month of January. 2. Mean Tb of Control black-tailed prairie dogs (36.2 degrees C) was significantly different from FWD (33.4 degrees C) and WD (30.4 degrees C) black-tailed prairie dogs. 3. VO2 was not significantly different between FWD and Control black-tailed prairie dogs (4.4 and 4.0 ml O2/kg/hr, respectively), while VO2 was significantly different between WD and Control animals (2.9 and 4.0 ml O2/kg/hr, respectively). 4. These findings are discussed as possible mechanisms for conserving body water.


Bile pigments in gallbladder and freshly-secreted hepatic duct bile from fed and fasted rainbow trout, Oncorhynchus mykiss.

1. Chromatographic analyses of bile pigments in rainbow trout reveal the presence of primarily unconjugated biliverdin (BV) and bilirubin (BR) glycosyl conjugates. Only trace amounts of unconjugated BR are present in hepatic duct (HD) bile: no beta-glucuronidase activity is detectable. 2. The per cent of BV and BR in HD and gallbladder biles is similar in fasted trout; however, the per cent of BV is significantly increased in HD bile from fed fish. 3. Fasting decreases the rate of choleresis but does not alter the excretory rate of endogenous BV or BR. 4. Erythrocyte life span is estimated to be approximately 500 days.


Stimulation of corticosterone secretion by dietary histidine in rats.

1. Liver glycogen accumulated within 3 days after the initiation of a histidine-excess diet. Serum corticosterone increased and serum insulin decreased, but plasma glucagon remained unchanged. 2. When 2 mmol (310 mg) of histidine was administered to fasted rats, serum corticosterone increased after 5 hr and tended to be higher 9 hr after administration. 3. Liver glycogen tended to accumulate after 5 hr and had accumulated significantly after 9 hr.


Digesta retention and fibre digestion in maras (Dolichotis patagonum) and guinea-pigs.

1. Digestibilities of feed and turnover time (1/k), transit time (TT) and mean retention time (MRT: 1/k+TT) of fluid and particle markers were measured in maras (Dolichotis patagonum) and guinea-pigs (Cavia procellus) fed a diet containing 50% alfalfa. 2. The digestibility of fibre was similar in both animals, however, the digestibilities of crude protein (nitrogen x 6.25) and crude ash were lower in the mara than in the guinea-pig. 3. 1/k of the digesta markers were similar in both animals, suggesting that the two animals possess similar dilution and retention time of digesta in their caecum and proximal colon.


Lipolytic and diabetogenic effects of native and biosynthetic growth hormone in the chicken: a re-evaluation.

1. There is controversy as to whether growth hormone (GH) is lipolytic and/or diabetogenic in vivo in chickens. The ability of GH to influence circulating concentrations of free fatty acids was examined in anaesthetized (suppressing endogenous GH secretion) adult and young chickens using three preparations of GH. 2. Plasma concentrations of free fatty acids were increased following the intravenous injection of native bovine GH (50 micrograms/kg to either young or adult chickens), recombinant chicken GH (American Cyanamid) (50 micrograms/kg to adult chickens) and recombinant chicken GH (Amgen) (50 micrograms/kg to young chickens). 3. Similarly, infusion of recombinant chicken GH was accompanied by a gradual increase (P less than 0.05, 90 min following start of infusion) in plasma concentrations of free fatty acids in both anaesthetized hypophysectomized and sham operated young chickens. 4. These data support an acute lipolytic role for GH in the chicken. 5. The injection of neither bovine nor chicken GH had any consistent effect on circulating concentrations of glucose. Moreover, if GH was administered in the presence of glucose, GH had no effect on plasma concentrations of glucose. 6. Further evidence for a lack of a diabetogenic role for GH comes from the inability of chronic administration of GH to influence the decline in plasma concentrations of glucose following challenge with the insulin secretagogue, tolabutamide.


A case of Graves' disease and papillary thyroid carcinoma with predominant production of thyroid-stimulation-blocking antibodies (TSBAb) persisted after total thyroidectomy.

A 42-year-old female with Graves' disease and papillary thyroid carcinoma with lung metastasis was referred to our hospital. After treatment of thyrotoxicosis with methimazole and Lugol's solution, she underwent total thyroidectomy. She was then given 131I twice to treat lung metastasis. However, 131I uptake into the lung was not clear in the scintigram. Both thyroid-stimulating antibodies (TSAb) and thyroid-stimulation-blocking antibodies (TSBAb) were detected in her sera before and after the treatments. Compared with TSAb activities, TSBAb activities were extremely high. Changes in the titers of these two antibodies were not clear after total thyroidectomy. These results indicate that lymphocytes outside the thyroid gland are the major source of TSAb and TSBAb in this patient.


A kindred of multiple endocrine neoplasia type 2B.

We describe familial cases of multiple endocrine neoplasia (MEN) 2B: A 48-year-old man is the proband. He had pheochromocytoma, medullary thyroid carcinomas (MTCs), parathyroid hyperplasia, mucosal neuromas, eversion of eyelids and Marfanoid appearance, and then underwent adrenalectomy and total thyroidectomy. Family screening revealed that his two daughters (10 and 8 years old) had mucosal neuromas and increased serum calcitonin (CT). Both of them had MTCs but no pheochromocytoma, and their MTCs were surgically removed. The father and his children have been in favorable condition since the operations. Southern blot analysis with 33 polymorphic DNA probes was done in MTCs obtained from two daughters. An RBP3 (10q11.2) locus linked to a predisposing gene on chromosome 10 was uninformative in either patient because of constitutional homozygosity. Loss of heterozygosity at the MYCL1 locus on chromosome 1p32 was observed in MTC from the younger sister, but no loss of heterozygosity was recognized in other loci examined. Deletion of the 1p32 locus may play a role in the development of MEN 2B.


The interactions of ethanol with single and repeated doses of suriclone and diazepam on physiological and psychomotor functions in normal subjects.

The effects of diazepam (5 mg t.i.d.), suriclone (0.2 and 0.4 mg t.i.d.) and placebo (t.i.d.) were assessed in 12 normal, healthy volunteer subjects after a single dose and after treatment for a period of 8 days. A battery of physiological, psychomotor and subjective tests was administered on days 1 and 8 both before and after drug and after a measured dose of ethanol. The effects of diazepam on the EEG were characteristic of benzodiazepines-a decrease in the slow frequency wave-bands, an increase in fast frequency wave-band and diminished evoked response amplitude. Suriclone had similar effects on fast frequency activity and evoked response amplitude but, in contrast to diazepam, also increased the slow frequency wave-bands after 7 days treatment. Some improvements in performance were seen with suriclone on critical flicker fusion, tapping speed, spiral maze and digit cancellation. In contrast, suriclone impaired performance to a greater extent than diazepam on digit symbol substitution and symbol copying. Body sway was also enhanced by suriclone to a greater extent than diazepam. Subjective ratings for mood and adverse-effects showed few differences between suriclone or diazepam. However, suriclone caused greater gastro-intestinal disturbances than diazepam, especially after ethanol, and subjects rated themselves as more antagonistic and more irritable on suriclone. Ratings for calmness suggested that in contrast to diazepam, suriclone had no anxiolytic effect. Several of the parameters tested showed a build up of effect with diazepam over the treatment period which was not seen with suriclone. It is suggested that this difference may be due to differences in elimination rate.(ABSTRACT TRUNCATED AT 250 WORDS)


Use of benzodiazepines and industrial injuries. Service Medical Interentreprises de Toulouse, France.

A one-year case-control study was performed by occupational physicians in 328 injures people and 662 controls in order to investigate a possible relationship between the consumption of benzodiazepines and the occurrence of industrial injuries. Benzodiazepine consumption was found to be related to sex (females) and age but not to a higher frequency of industrial injuries. It is suggested that knowledge of being at risk might be a factor reducing the consumption of benzodiazepines.


The effects of the normal and oncogenic forms of the neu tyrosine kinase, and the corresponding forms of an immunoglobulin E receptor/neu tyrosine kinase fusion protein, on Xenopus oocyte maturation.

In this work, we have used Xenopus oocyte maturation as a read-out for examining the ability of the neu tyrosine kinase (p185neu) to participate with the epidermal growth factor (EGF) receptor in a common signal transduction pathway. We find that unlike the case for the EGF receptor, which elicits EGF-dependent maturation of these oocytes as reflected by their germinal vesicle breakdown (GVBD), neither the normal neu tyrosine kinase (p185val664) nor the oncogenic form of neu (p185glu664) are able to effectively trigger this maturation event. However, expression of p185glu664 causes a specific and significant promotion of the progesterone-induced GVBD, reducing the half-time for this maturation even from approximately 9 h to approximately 5 h. Stimulation of the progesterone-induced GVBD did not occur following the expression of a kinase-deficient p185neu protein (in which a lysine residue at position 758 was changed to alanine). Essentially identical results were obtained when the mRNAs coding for fusion proteins comprised of the extracellular domain of the receptor for immunoglobulin E (IgE), and the membrane-spanning and tyrosine kinase domains of normal or oncogenic p185neu (designated IgER/p185val664 and IgER/p185glu664, respectively), were injected into oocytes. Antigen-induced crosslinking of IgER/p185val164 proteins expressed in oocytes caused a reduction in the half-time for the progesterone-stimulated GVBD from approximately 9 h to approximately 7 h. Thus, the aggregation of the membrane-spanning and/or tyrosine kinase domains of p185val664 partially mimics the effects of the oncogenic forms of p185neu. Overall, the results of these studies suggest that the activation of the p185neu tyrosine kinase by a point mutation within its membrane-spanning helix, or an aggregation event, can result in the facilitation of oocyte maturation events that are elicited by other factors (e.g. progesterone). However, the activated p185neu tyrosine kinases are not able to mimic the EGF-stimulated EGF receptor tyrosine kinase in triggering oocyte maturation, which suggests that the EGF receptor and the p185neu tyrosine kinase do not input into identical signal transduction pathways in these cells.


Effects of quinpirole on autonomic nervous control of heart rate in rats.

The effects of quinpirole, a specific dopamine DA2 receptor agonist, on autonomic nervous control of heart rate, were studied in normotensive pithed rats, by analysing its action on the tachycardia and bradycardia evoked by electrical stimulation of the cardioaccelerator (10 V; 1 ms; 0.5, 1, 3, 6 Hz) and vagus (10 V; 1 ms; 3, 6, 9 Hz) nerves respectively. Quinpirole (10-50-100 micrograms kg-1 iv) reduced the cardioacceleration elicited by electrical stimulation but not that by noradrenaline (3 micrograms kg-1 iv). The effect on electrical stimulation was blocked by domperidone (0.5 mg kg-1 iv) but not by SCH 23390 (0.2 mg kg-1 iv) or idazoxan (0.3 mg kg-1 iv). At 1, 5, 10, 30 micrograms kg-1 iv, quinpirole decreased vagal but not acetylcholine-induced bradycardia. The effect on electrical stimulation was inhibited by domperidone (0.5 mg kg-1 iv) but not by SCH 23390 (0.2 mg kg-1 iv), prazosin (0.1 mg kg-1 iv) or idazoxan (0.3 mg kg-1 iv). The data point to the presence of presynaptic and/or ganglionic dopamine receptors in the sympathetic and parasympathetic innervation of the rat heart, where stimulation inhibits the release of neuromediators.


Expression of proliferating cell nuclear antigen in lung cancer: a systematic study and correlation with DNA ploidy.

Heterogeneity of expression of the proliferating cell nuclear antigen (PCNA) was assessed immunohistochemically in 156 tissue samples from 33 surgically resected pulmonary carcinomas using the monoclonal antibody 19A2. The DNA content of each of these samples was measured by flow cytometry. Mean PCNA expression was higher in squamous carcinomas than in adenocarcinomas but there was marked intra-tumour variation in PCNA index in almost all cases. Intra-tumour heterogeneity of DNA content was noted in 11 cases. The PCNA index of these cases (34.1) was higher than that of DNA homogeneous cases (19.4). The wide variation in PCNA expression between different samples within a tumour would indicate that systematic sampling and counting will be necessary in future immunohistochemical studies of cell proliferation in tumour material.


Neuroleptic malignant syndrome: description of two cases with prolonged clinical course.

Two cases of neuroleptic malignant syndrome presenting an unusual clinical course are reported. The first patient was untreated for the syndrome and recovered completely only after four months, while the other one was given dopaminergic and myorelaxing drugs only 10 days after the onset of the symptoms and died about six months later with an unmodified clinical picture. In both cases the treatment seemed to influence the clinical course, a delay or lack of drug intake worsening the prognosis.


American Indian prehistory as written in the mitochondrial DNA: a review.

Native Americans have been divided into three linguistic groups: the reasonably well-defined Eskaleut and Nadene of northern North America and the highly heterogeneous Amerind of North, Central, and South America. The heterogeneity of the Amerinds has been proposed to be the result of either multiple independent migrations or a single ancient migration with extensive in situ radiation. To investigate the origin and interrelationship of the American Indians, we examined the mitochondrial DNA (mtDNA) variation in 87 Amerinds (Pima, Maya, and Ticuna of North, Central, and South America, respectively), 80 Nadene (Dogrib and Tlingit of northwest North America and Navajo of the southwest North America), and 153 Asians from 7 diverse populations. American Indian mtDNAs were found to be directly descended from five founding Asian mtDNAs and to cluster into four lineages, each characterized by a different rare Asian mtDNA marker. Lineage A is defined by a HaeIII site gain at np 663, lineage B by a 9-bp deletion between the COII and tRNA(Lys) genes, lineage C by a HincII site loss at np 13259, and lineage D by an AluI site loss at np 5176. The North, Central, and South America Amerinds were found to harbor all four lineages, demonstrating that the Amerinds originated from a common ancestral genetic stock. The genetic variation of three of the four Amerind lineages (A, C, and D) was similar with a mean value of 0.084%, whereas the sequence variation in the fourth lineage (B) was much lower, raising the possibility of an independent arrival. By contrast, the Nadene mtDNAs were predominantly from lineage A, with 27% of them having a Nadene-specific RsaI site loss at np 16329. The accumulated Nadene variation was only 0.021%. These results demonstrate that the Amerind mtDNAs arose from one or maybe two Asian migrations that were distinct from the migration of the Nadene and that the Amerind populations are about four times older than the Nadene.


Herpetic stromal keratitis: an immunopathologic disease mediated by CD4+ T lymphocytes.

To investigate the role of T cell subsets in the development of herpetic stromal keratitis (HSK) in a well defined model, we used an adoptive transfer approach in which thymectomized and T cell-depleted mice [T(-)] were reconstituted with different numbers of syngeneic immune T lymphocytes after topical corneal challenge with RE strain of herpes simplex virus-1. In vitro stimulated or unstimulated immune T cells obtained from cervical and retropharyngeal lymph nodes of mice with HSK were used in adoptive transfer experiments. Although T(-) mice developed an initial epithelial inflammation, stromal keratitis did not occur. Reconstitution experiments revealed that mice that received 2 x 10(7) or more unfractionated immune T cells could develop HSK lesions with severity comparable to immuno-competent control mice. In mice receiving CD8(+)-depleted populations, even fewer cells (5 x 10(6)/mouse) were able to induce significant HSK. In contrast, mice that received similar or increased numbers of cells depleted of CD4+ T lymphocytes did not develop HSK. Immune T lymphocytes transferred to mice that were mock infected on the cornea did not develop HSK, indicating that the immunopathogenic cells were virus specific and not merely reacting to autoantigens. Histopathologic examination of the diseased corneas demonstrated that the stromal inflammation in euthymic normal and T(-)-reconstituted mice was characterized by extensive polymorphonuclear leukocyte infiltration. Scattered lymphocytes, and occasional macrophages also were observed. These results provide further evidence that HSK represents an immunopathologic process mediated mainly by CD4+ T cells.


Corneal epithelial glycoproteins exhibit Pseudomonas aeruginosa pilus binding activity.

Adherence of Pseudomonas aeruginosa to the cornea is a requisite step in the pathogenesis of bacteria-induced corneal disease. P. aeruginosa is capable of attaching to host epithelial cells by its pili, but there is little information regarding the epithelial receptors of this adhesin in the cornea. Using nitro-cellulose blotting of polyacrylamide gels of solubilized adult mouse corneal epithelium, four major proteins (molecular weights: 38, 42, 57, and 66 kD) and several minor proteins were identified that bound purified pili from strain PAK and its hyperpiliated mutant PAK/PR1. These proteins were identified by immunoblotting either with pilus-specific monoclonal antibodies, XLR-3 and PK 3B, or using peptide PAK 128-144 (OX). The glycosylated nature of the proteins was determined using similar gel electrophoresis of corneal epithelial proteins, blotting onto nitrocellulose, and staining the blots with lectins conjugated to either horseradish peroxidase or alkaline phosphatase. All four major pilus-binding proteins were stained with concanavalin A lectin (mannose and glucose) and either wheat germ agglutinin lectin (WGA, specific for sialic acid and N-acetylglucosamine) or succinylated WGA lectin (only N-acetylglucosamine). Staining for peanut agglutinin lectin (galactose beta(1-3) N-acetylgalactosamine) was seen for the 42-, 57-, and 66-kD proteins. The importance of the carbohydrate portions of these corneal proteins in pili binding was confirmed by preincubation of corneal epithelial blots with periodate or pili with sialic acid, both of which abolished the pili binding. These studies indicate that corneal epithelial pilus-binding proteins are glycoproteins in nature and that sialic acid may be a constituent of these pilus-specific receptors in the adult mouse corneal epithelium.


Re-examination of peripheral blood T cell subsets in dysthyroid orbitopathy.

Patients with dysthyroid orbitopathy (DO) were grouped according to a multifactorial assessment of disease severity and the rate of disease progression. Using this system and flow cytometric measurements of T cell subsets in the peripheral blood, a significant increase in the percentage of CD4+ lymphocytes correlated with disease severity in DO patients with progressive disease. These observations are consistent with the hypothesis that the CD4+ peripheral blood T helper cells play a significant role in the progression of DO.


Cobalamin deficiency associated with methylmalonic acidemia in a cat.

A 9-month-old sexually intact male longhair cat was examined because of lethargy, anorexia, cold intolerance, and failure to thrive since acquisition at an early age. Clinical signs of disease were less pronounced when the cat was fed a low-protein diet. Anemia, hypoglycemia, low total CO2 content, and hyperammonemia were detected. The cat was euthanatized. Urine obtained immediately before euthanasia contained a large amount of methylmalonic acid. Total serum cobalamin concentration was low. Hepatic methylmalonic-CoA mutase activity, with and without the addition of coenzyme adenosylcobalamin, was consistent with a cobalamin deficiency. Methylmalonic acidemia secondary to a putative defect in cobalamin absorption was diagnosed.


Cryptorchidism and monorchism in cats: 25 cases (1980-1989).

Of 1,345 cats admitted for orchiectomy during a 10-year period, 23 (1.7%) were cryptorchid and 2 (0.1%) were monorchid. Persian cats were over-represented in the cryptorchid population (P = 0.01). Cats were more likely to be unilaterally than bilaterally cryptorchid (P = 0.01). A predisposition for location of undescended testes (abdominal vs inguinal or right vs left side) was not identified in unilateral cryptorchids. All bilateral cryptorchids had abdominally located testes. The most common surgical approaches used for orchiectomy of cryptorchid cats were a caudal ventral midline incision for inguinal testes and a caudal ventral midline celiotomy for abdominal testes.


Novel restriction fragment length polymorphisms in the cellular oncogene SEA.

The human homologue of the SEA oncogene has been mapped recently to chromosome band 11q13. While studying the possible involvement of this gene in the variant translocation t(9;22;11) (q34;q11;q13) in a case of chronic myelogenous leukemia, we identified novel polymorphisms for XbaI and SacI restriction enzyme sites in the SEA gene. Frequency of the polymorphic alleles was studied in 100 samples from healthy controls, 94 samples from patients with non-Hodgkin's lymphoma, 25 samples from patients with benign lymphadenopathy, and 38 samples from patients with chronic myelogenous leukemia. XbaI digestion showed a three-allele polymorphism with two frequent alleles A (8.0 kb) and B (9.2 kb) and a rare allele (5.8 kb). After SacI digestion the probe identified two primary genotypes. Genotype I showed two hybridizable DNA fragments, one each of 6.6 and 3.5 kb. In genotype II the 3.5 kb fragment was absent, instead two smaller fragments, one each of 1.9 kb and 1.6 kb were present. The 6.6 kb fragment (allele AA) had three polymorphic sites generating 6.2 kb fragment (allele BB), 7.4 kb fragment (allele CC), and 7.8 kb fragment (allele DD). Frequencies of the two genotypes and the four alleles followed Mendelian proportions in all the samples studied. Furthermore, this study shows the importance of restriction map analysis of DNA in the vicinity of the probe of an oncogene to distinguish natural polymorphisms from the disease-related rearrangements in the gene.


Diagnostic confusion in treatment-refractory psychotic patients.

Two surveys of diagnostic practices in the United States suggest that many clinicians base their diagnoses on presenting symptoms and pay little attention to course and exclusionary criteria. Failure to correctly diagnose patients may result in inappropriate therapy and poor treatment response. The purpose of the present study was to investigate diagnostic practices. We made detailed assessments of 50 consecutively admitted treatment-refractory psychotic patients and carefully applied DSM-III-R criteria. Referral diagnoses were changed in 23 of the 50 patients. Diagnoses of schizophrenia and schizoaffective disorder were made far less frequently and mood disorders (bipolar disorder and major depression) were diagnosed far more frequently by our group than by referring psychiatrists. Patients whose diagnosis was changed were more likely to be given mood-stabilizing medication and tended to show more improvement than patients whose diagnosis was not changed. These findings raise the possibility that patients may not respond to treatment because incorrect diagnoses result in inappropriate treatment.


Nutritional and hormonal regulation of mRNA levels of lipogenic enzymes in primary cultures of rat hepatocytes.

The effects of nutrients and hormones on the mRNA levels of acetyl-CoA carboxylase, fatty acid synthase, malic enzyme, and glucose 6-phosphate dehydrogenase were examined in primary cultures of rat hepatocytes during the process of induction. The addition of both glucose and insulin to the culture medium markedly enhanced the lipogenic enzyme mRNA induction due to either of them, in 16 h. Fructose or glycerol proved to be an effective substitute for glucose, suggesting that glycolytic metabolites were involved in the mRNA induction. It is remarkable that mRNA induction of acetyl-CoA carboxylase was the most sensitive to glucose and also to insulin among the lipogenic enzymes. Polyunsaturated fatty acids markedly reduced the mRNA induction of lipogenic enzymes. Dexamethasone enhanced all the lipogenic enzyme mRNA induction by insulin. On the other hand, triiodothyronine addition greatly increased the mRNA concentrations of lipogenic enzymes, but dexamethasone decreased rather than increased the mRNA induction by triiodothyronine. The effects of insulin on the induction of the lipogenic enzyme mRNAs were similar, but those of triiodothyronine were not. Triiodothyronine markedly enhanced malic enzyme mRNA induction by insulin with dexamethasone, and tended to enhance the induction of the acetyl-CoA carboxylase and fatty acid synthase mRNAs, but not that of glucose 6-phosphate dehydrogenase mRNA. It appeared that insulin and triiodothyronine synergistically enhanced lipogenic enzyme mRNA induction by glucose, but the mechanisms were different.


Restriction fragment length polymorphism analysis of the fimbrillin locus, fimA, of Porphyromonas gingivalis.

With hybridization probes derived from the fimbrial locus of Porphyromonas gingivalis strain 381, fimA381, restriction fragment length polymorphisms (RFLPs) were examined at the fimbrillin locus in 39 human and animal strains of this species. The 39 strains were subdivided into nine RFLP groups (I-IX) after genomic digests were probed with the internal coding sequence of the fimA381 gene. Thirty-three strains showed one or more AluI fragments of moderate-to-high homology (greater than or equal to 77%) with the internal coding sequence of fimA381. These strains were distributed into the first seven RFLP groups, based solely on the size of the major hybridizing AluI fragment. Five human strains (RFLP Group VIII) had only one AluI fragment that hybridized very poorly with this probe. One animal strain did not have homology at all (RFLP Group IX). When all AluI fragments that hybridized with fimA381 were analyzed, RFLP groups I-VIII were further differentiated into 25 distinct RFLP patterns. Hybridizations were also performed with the internal coding sequence of fimA381 to probe PstI genomic digests of selected strains that appeared to have lesser homology with fimA381. These hybridizations were performed to determine the level and location of the region of poor homology within the fimA genes of these strains. The results suggested that fimbrial coding sequences are more commonly conserved between these strains in the 5'-region of the fimA locus (greater than or equal to 92% sequence homology).(ABSTRACT TRUNCATED AT 250 WORDS)