A T cell/B cell/epithelial cell internet for mucosal inflammation and immunity

Conclusions Mucosal immune responses are strongly regulated by CD4⁺ T cells and their derived cytokines. In this regard, IFN-^–/– mice (i.e., which lack Th1 and have elevated Th2 cells) showed strong mucosal Th2-type responses together with S-IgA production, while IL-4^–/– (e.g., dominant Th1 and lack of Th2 cells) mice had impaired mucosal Th2 and IgA responses following oral delivery of TT and CT. However, when rSalmonella or radenovirus were used for antigen delivery, significant levels of mucosal IgA responses were induced in both IFN-^–/^– and IL-4^–/^– mice. The choice of the antigen delivery system which leads to optimal Th and B cell interactions are important for the induction of effective IgA responses, even in situations where the immune system is compromised. It is clear that Th2-type cytokines are important in mucosal IgA responses; however, other cytokine combinations can compensate for mucosal immunity in situations in which Th2 cell responses are absent. Mucosally induced tolerance may be one approach to prevent several systemic immune disorders; however, the mechanism of this phenomenon still needs to be elucidated. Our recent findings have suggested that IFN- may play an important role in induction of systemic unresponsiveness since oral tolerance was not induced in IFN-^–/^– mice.Our studies as well as those of others indicated that at least two phases of a triad of cell interactions are important for the mucosal immune system. First, it has been shown that epithelial cell-produced IL-7 and SCF and T cell-derived IL-2 are essential activation and growth signals for intestinal T cells. Second, our studies with TCR knockout mice have suggested that mucosal T cells also play a critical role in the regulation of mucosal IgA responses. Thus, a mucosal internet among T cells, T cells, and IgA B cells appear critical for mucosal homeostasis and for regulation of specific mucosal immune responses.     

TGF-β1 Null Mutation Leads to CD154 Upregulated Expression in Affected Tissues

The TGF-1(–/–) mouse is a murine model for systemic autoimmune disease. The aim of this study is to elucidate the immunological mechanism that leads to multifocal tissue inflammation and autoantibody production in TGF-1(–/–) mice. Heart, lung, liver, and salivary gland from TGF-1(–/–) were assessed for CD154 expression by RT-PCR and immunohistochemistry. Compared to wild-type littermates, CD154 expression was elevated in all tissues studied. Furthermore, IL-12 mRNA was expressed in the salivary gland and heart of TGF-1(–/–) mice and not in wild-type littermates. This suggests that the CD154 pathway is activated in these tissues. This shows that TGF-1 regulates CD154 expression leading to spontaneous IL-12 production and autoimmunity.     

„Leptospira Mini“ ein neuer Serotyp pathogener Leptospiren

Zusammenfassung Auf Grund ihres immunbiologischen Verhaltens gehören die Leptospirenstämme Sari, Ghidorsi und Szwajizak zu demselben serologischen Leptospirentyp, für den der Name Leptospira Mini vorgeschlagen wird.Der Stamm Sari wurde 1942 vonMino sowieVercelli in Italien isoliert. Der Stamm Ghidorsi wurde von uns im Zuge unserer Leptospirenforschungen bei einer Reisfeldarbeiterin der Po-Ebene nachgewiesen. Der Stamm Szwajizak, der vonSmith, Brown, Tonge u. Mitarb. im Jahre 1954 beschrieben wurde, ist in Nord-Qeensland gefunden worden. Der Stamm Sari und Ghidorsi gehören dem kompletten Biotyp (AB), der Stamm Szwajizak dem inkompletten (A) an.Leptospira Mini gehört zur Serogruppe hebdomadis. Ihre Virulenz ist schwach und ihre Bedeutung als Erreger menschlicher Leptospiren-infektionen scheint gering zu sein.     

Use of B-IV zonal rotor centrifugation as a simple tool for the separation of adeno-associated X7 virus (AAV X7) from helper adenoviruses

Summary The use of zonal rotor centrifugation in the separation of AAV X₇ virus from its helper adenovirus is reported. AAV X₇ virus was perfectly separated from its helper adenovirus. Adeno-associated X₇ virus was found to separate in par tides with an S rate of 72–82 S, and a density in CsCl of 1.30–1.32, and virions with a sedimentation coefficient of 135–147 Svedbergs, with a density of 1.37–1.38.     

Triggering of respiratory burst by tumor necrosis factor in neutrophils adherent to fibronectin. Evidence for a crucial role of CD18 glycoproteins

Human neutrophils, plated on fibronectin (FN)-coated wells, were found to release large quantities of superoxide anion (O ₂ ^– ) in response to tumor necrosis factor alpha (TNF-). The O ₂ ^– release was completely inhibited by two monoclonal antibodies (MoAbs, MHM23 and TS1/18) against CD18 glycoproteins. An independently derived anti-CD18 MoAb (60.3) was ineffective. These MoAbs failed to inhibit neutrophil adhesion to FN-coated surfaces. Moreover, neutrophils incubated for 30 min on FN and then washed to remove non-adherent cells, were responsive to TNF- in the presence of anti-CD18 MoAbs MHM23 and TS1/18. Consequently, the CD18-dependent capacitation of the respiratory burst can occur before TNF- triggering. Finally, neutrophils plated on FN in the presence of anti-CD18 MoAbs and then washed, i.e. adherent cells blocked in their surface CD18 molecules, released O ₂ ^– after adding TNF- but only in the absence of additional anti-CD18 MoAbs. This is consistent with a TNF- ability to induce rapid expression and activation of new oxidative burst-capacitating CD18 molecules. The results suggest that the anchorage of neutrophils to FN surfaces depends on adherence molecules apparently unrelated to CD18, probably the so-called fibronectin receptors (FNRs), whereas the capacitation of the respiratory burst in response to TNF- requires the intervention of CD18 glycoproteins, available on the membrane of resting neutrophils or mobilized to the cell surface by TNF-.     

Evidence for Na+/Ca2+ exchange in isolated smooth muscle cells: a fura-2 study

Isolated smooth muscle cells (SMC) from guinea pig taenia coli were employed. Suspension of cells were externally loaded in saline with the fluorescent calcium indicators quin-2/AM or fura-2/AM at 20–40 M or 4 M respectively, resulting in an estimated intracellular concentration of 100–200 M for quin-2 or 10–20 M fura-2 (free acid). On addition of 100 M carbachol or high K _o ⁺ (80 mM) depolarization, fura-2 loaded cells contracted (104±47 m,n=121 rest: 39±13 m,n=59 contracted) identically to control (103±35 m,n=232 rest: 39±16 m,n=89 contracted) cells, whereas quin-2 loaded cells were unresponsive to these protocols and there was no significant length change. The Ca _i ²⁺ of fura-2 loaded cells was 100±18 nM (mean±SD,n=15) and was not significantly different from quin-2 loaded cells 107±26 nM (n=13). Treatment of fura-2 loaded cells with 100 M ouabain saline for 10–60 min progressively elevated the Ca _i ²⁺ to a mean of 266±83 nM (n=15). Reduction of Na _p ⁺ (96% Li⁺ replaced) significantly increased Ca _i ²⁺ to 317±77 nM (n=8). After pretreatment with ouabain (100 M), Na _o ⁺ replacement (Li⁺) increased Ca _i ²⁺ at a significantly faster rate [3.6 nM min^–1 (control) cf. 19.8 nM min^–1 (ouabain)].     

Skull radiograph measurements of normals and patients with basilar impression; use of Landzert's angle

Summary One hundred normal lateral skull radiographs were studied and those of ten patients with basilar impression attending Kenyatta Hospital, Nairobi. The mean shortest distance of the odontoid tip to McGregor's basal line was 1.2±2.28 mm below the basal line (range 6 mm below to 3 mm above basal line), in normals and 9±2.7 mm (6–14 mm) above basal line in patients. The mean basal angle was 113±7 (102–133) in normals and 122±6 (113–125) in patients. The mean nasion-basion-opisthion angle was 162±4 (154–169) in normals and 178±5 (173–185) in patients. The mean total length of clivus was 48±3.7 mm (43–56 mm) in normals and 44±6.6 (36–48 mm) in patients group. The mean median diameter of the foramen magnum was 39±5 mm (30–48 mm), atlas 21±3 mm (18–25 mm) axis 18±3 mm (14–23 mm), third cervical vertebra 16±2 mm (13–22 mm) in normals and in patients: 39±4 mm (36–45 mm), atlas 23±6 (l5–30 mm) axis 19±4 mm (16–25 mm), third cervical vertebra 16±3 (14–20). There was a significant difference in the position of the odontoid tip and the nasion-basion-opisthion angle between the normal and patient groups. All the other parameters measured in this work did not differ significantly between the two groups.

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Etude anatomo-radiologique de crânes normaux et de crânes pathologiques avec impression basilaire; utilisation de l'angle de Landzert

Résumé Cent crânes normaux ont été étudiés sur des radiographies de profil ainsi que dix crânes pathologiques présentant des impressions basilaires chez des patients traités à l'HÔpital Kenyatta de Nairobi. La plus courte distance moyenne entre le sommet de l'odontoÏde et la ligne basale de McGregor a été de 1,2±2,28 mm au-dessous de la ligne basale (extrÊmes étendues de 6 mm au-dessous à 3 mm au-dessus de la ligne basale), chez les sujets normaux et de 9±2,7 mm (6–14 mm) au-dessus de la ligne basale chez les sujets pathologiques. L'angle basai moyen était de 113±7 (102–133) chez les sujets normaux et 122±6 (113–125) chez les sujets pathologiques. L'angle moyen nasion-basion-opisthion était de 162±4 (154–169) chez les sujets normaux et 178±5 (173–185) chez les sujets pathologiques. La longueur moyenne totale du clivus était de 48±3,7 mm (43–56 mm) chez les sujets normaux et 44±6,6 (36–48 mm) chez les sujets pathologiques. Le diamètre moyen du foramen magnum était de 39±5 mm (30–48 mm), celui du foramen vertébral de l'atlas était de 21±3 mm (18 à 25 mm), celui de l'axis (18±3 mm (14–23 mm), celui de la troisième vertèbre cervicale: 16±3 mm (13–22 mm) chez les sujets normaux; chez les sujets pathologiques les chiffres étaient les suivants: foramen magnum 39±4 mm (39–45 mm), atlas 23±6 (15–30 mm), axis 19±4 mm (16–25 mm), troisième vertèbre cervicale 16±3 mm (14–20 mm). Il existe une différence significative dans la position du sommet de l'odontoÏde et la valeur de l'angle nasion-basion-opisthion entre les deux groupes. Aucun des autres paramètres mesurés dans ce travail ne présentait de différence significative entre les deux groupes.

     

”Reflexes of purpose and freedom” in the comparative physiology of higher nervous activity

The most complex unconditioned reflexes of aim and freedom, discovered by I. P. Pavlov, are compared with the competence drive and the motivation of the resistance to coercion, respectively, described by contemporary ethologists. On the basis of the unconditioned reflex of purpose, conditioned reflexes were developed in which positive emotions arising in connection with the perfection of a skill, irrespective of its pragmatic significance at a given moment, serve as the reinforcement. The unconditioned reflex of freedom is regarded as a phylogenetic precursor of the will, and its acute extinction as the physiological mechanism of hypnosis. It was demonstrated experimentally that the appearance of the state of animal hypnosis (immobilization catatonia) in rabbits is accompanied by the predominance of electrical activity and heat production in the right hemisphere, i. e., by symptoms which are found in hypnosis in man.Translated from Zhurnal Vysshei Nervnoi Deyatel'nosti imeni I. P. Pavlova, Vol. 39, No. 3, pp. 415–420, May–June, 1989.     

Nephritogenicity and α-chain composition of NC1 fractions of type IV collagen from bovine renal basement membrane

Nephritogenicity (anti-GBM-nephritis-inducing activity) and -chain composition of globular-domain (NC1) fractions of type IV collagen from bovine renal, pulmonary, and placental basement membranes (BMs) was examined by injecting these fractions with adjuvant into WKY/NCrj rats and by Western blotting using epitope-defined monoclonal antibodies to the six different chains of type IV collagen. A purified nephritogenic fraction from renal BM contained 1–6(IV)NC1, whereas a non-nephritogenic fraction contained only 1–2(IV)NC1. Renal and pulmonary NC1 had strong nephritogenic activity; placental NC1 had weak activity. The renal and pulmonary fractions contained 1–6(IV)NC1, and the placental fraction had a large amount of 1–2(IV)NC1 and a very small amount of 3–6(IV)NC1. Immunohistochemical study of bovine renal BM with the monoclonal antibodies revealed that bovine glomerular BM contained 1–5(IV) chains, but not the 6(IV) chain. The absence of 6(IV) chain in glomerular BM in bovine and in humans indicates that 6(IV) chain is not a target antigen of anti-GBM nephritis. Nephritogenicity is apparently a property of 3–5(IV)NC1.     

Veratridine-induced high-frequency asynchronous release of inhibitory transmitter quanta in crayfish nerve-muscle synapses superfused with normal and low-calcium saline

Crayfish fibres of opener muscles were voltage clamped toE=–80 mV membrane potential (T=19–22°C), and veratridine (10–100 mol/l) was added to the superfusate. Within 30–60 s this caused large fluctuations of the clamp current due to vigorous asynchronous quantal release from the inhibitory nerve terminals along the muscle fibre. Excitatory postsynaptic receptors were previously desensitized by application of 5 mmol/l glutamate. Current fluctuations were evaluated by means of the noise analysis technique. Typically, 100 mol/l veratridine increased instantaneously the quantal release rate ñ from ñ<1 quantum/s toñ10,000 quanta/s. Thereafter, ñ declined exponentially with a time constant of 70s. On average, about 500,000 inhibitory quanta could be liberated in this way from the terminals on a single muscle fibre of 1 mm length. Serotonin (1 mol/l) facilitated the effect of lower veratridine concentrations (1–10 mol/l). In opener muscles veratridine-induced asynchronous quantal release showed little dependence on the bath concentration of Ca²⁺. The opposite was found for fibres of the superficial abdominal extensor muscle. Beside postsynaptic current fluctuations, veratridine elicited slowly changing average postsynaptic DC-currents which could be explained partly by superposition of individual inhibitory quantal currents. These DC-currents suggest that beside inhibitory quantal release another factor activates inhibitory postsynaptic receptors after application of veratridine.This investigation was supported by the Deutsche Forschungsgemeinschaft, SFB 220     

Intratypic antigenic heterogeneity of Coxsackie B viruses

Summary Neutralization tests, employing the cytopathogenic effect in tissue culture tubes, with a variety of homotypic antisera and strains of Coxsackie B viruses often yielded high titers in early readings and low titers in late readings — the break-through phenomenon — and occasionally also low, early-reading titers with heterologous, homotypic sera, which gave high titers with the homologous strains. Of 27 strains of Goxsackie B 1 to B 5, that were tested, 10 showed no break-through tendency while others showed varying degrees of break-through, without reference to any evidence of intratypic antigenic variation. There was a positive correlation between a small number of tissue culture passages away from man or mouse brain and the break-through tendency. Moreover, strains without break-through tendency yielded viral populations with marked break-through properties after a single intracerebral passage in newborn mice, and even after two subsequent tissue culture passages. Plaque-purified progeny exhibited the break-through phenomenon to the same extent as the original, unpurified cultures.The early readings yielded reproducible titers, which could be used for analysis of antigenic variation. Prime antigenic variants, of broader antigenic constitution than their non-prime relatives, were found among the Coxsackie B 2, B 3, and B 4 strains that were tested. These prime strains (e. g., B. V. A. 96- B2; Stevens - B 3, and Burrier or J. V. B. - B 4) were found to be antigenically broader than the prototype strains (Ohio 1 - B 2, Nancy - B 3, Powers or Texas 13 - B 4) generally used for the preparation of diagnostic antisera. The broader antigenicity of the prime variant was also present in plaque-purified progeny.Aided by grants from The National Foundation, Inc.The work reported here was carried out in 1957–1958 during Dr.Wigand's tenure of a fellowship in the Cincinnati Laboratory.     

The sensitivity to Zn2+ discriminates between typical and atypical mast cells

The effects of zinc gluconate have been studied on rat peritoneal mast cells and rat basophilic leukemia cells (RBL 2H3) stimulated by various secretagogues. The IC₅₀'s of zinc gluconate on peritoneal cells were (M)1.6, 1.9, 5.4 and 18 for ionophore A23187, phorbol 12-myristate 13-acetate, substance P and immunoglobulin E-antigen, respectively. Higher concentrations of zinc gluconate were required to inhibit histamine secretion from RBL 2H3 cells, i.e. 12 M (ionophore A23187) and 140 M (immunoglobulin E-antigen). Zinc gluconate (10^–4 to 10^–3 M) also inhibited the IgE-dependent contraction of guinea pig trachea but was unable to affect that induced by exogenous histamine. These results suggest that zinc gluconate acts intracellularly and is selective of typical or connective tissue mast cells.     

Untersuchungen Mit J131-Markiertemβ-Globulin Zur Frage Der Abstammung Der Liquoreiweisskörper

Zusammenfassung Die Abstammung der-Globuline im Liquor wurde bei 20 Fällen mit den verschiedensten neurologischen Erkrankungen untersucht. — Die spezifische Aktivität der-Globuline war bei normalen und pathologischen Liquors ausnahmslos niedriger als im Serum. Es treten demnach nur einzelne Serum--Globuline in den Liquor über, ein verschieden großer Anteil der Liquor--Globuline wird im Liquorraum gebildet. Die-Fraktion im Liquor besitzt einen Serumanteil, von dem die liquoreigenen oder auch cerebrogenen-Globuline unterschieden werden können. Beziehungen zwischen der Höhe des liquoreigenen-Globulinanteils zu einzelnen Krankheitsgruppen waren nicht herzustellen. Es ließ sich aber zeigen, daß eine Erhöhung des elektrophoretisch ermittelten relativen-Globulingehaltes im Liquor bei pathologischen Fällen nicht — wie bisher angenommen wurde — mit einer Zunahme des cerebrogenen Eiweißes einherzugehen braucht. — Die Bedeutung des liquoreigenen-Globulins ist noch unbekannt, auch ist es nicht möglich zu entscheiden, welche einzelnen Proteine innerhalb der-Fraktion im Liquorraum entstanden sind oder aus dem Serum stammen.Mit Unterstützung der Deutschen Forschungsgemeinschaft.     

Susceptibility to beta-lactam antibiotics in septicemia isolates from twenty-nine European laboratories

In 1984 the European Study Group on Antibiotic Resistance (ESGAR) consecutively collected gram-negative bacilli and staphylococci blood isolates and performed susceptibility testing with 11 antibiotics using the microdilution method. In all 2,578 isolates were collected: 68% gram-negative bacilli and 32% staphylococci. The MICs of ampicillin and cefazoline for the susceptible gram-negative bacilli were 1–8g/ml; of piperacillin0.5–4; of Sch 34343, cefotaxime, moxalactam, ceftazidime and aztreonam0.5–2g/ml; of cefoxitin, cefuroxime and cefamandole0.5–8g/ml. For susceptible staphylococci the MICs of cefazoline and cefuroxime were0.5–1g/ml, and of cefoxitin, moxalactam, ceftazidime and cefotaxime,0.5–32 g/ml. The resistance levels varied between laboratories and countries, being lower in Northern Europe. In clinical protocols on patients with gram-negative septicemia from whom cefazoline-resistant strains were isolated, cefotaxime was the beta-lactam most commonly used (12%). In protocols on patients with staphylococcal septicemia from whom gentamicin-resistant or cefazoline-resistant strains were isolated, the most commonly used beta-lactam was cloxacillin (6%).     

Anti-oncospheral antibodies in the serum of lambs experimentally infected with eitherTaenia ovis orTaenia hydatigena

Using a peroxidase micro-enzyme-linked-immunosorbent assay (ELISA) method anti-oncospheral antibodies were demonstrated in sera from four lambs after primary and challenge infections withTaenia ovis orT. hydatigena. Antibodies assayed using homologous oncospheral antigen (OA) reached a peak by 3–4 weeks after primary infection and at 1–3 weeks post-challenge infection, but waned to pre-infective, background levels by 8–12 weeks after each infection. Antibodies assayed against antigens in strobilar or cystic larval extracts persisted for long periods after the initial infections and exhibited different kinetics of response from those demonstrated against OA. These antibodies showed increased levels after challenge infection. Oncospheral antigens did not seem to be species specific although they appeared to elicit a stage-specific response. It is suggested that the anti-oncospheral antibody response could be associated with protective or functional antibody.     

Some properties of the UL-FS 49 block of the hyperpolarization-activated current (i f) in sino-atrial node myocytes

Block of the hyperpolarization-activated pacemaker current (i _f) by the bradycardic agent UL-FS 49 was studied in isolated sino-atrial (SA) node myocytes. Using repetitive activation/deactivation protocols, micromolar concentrations of UL-FS 49 blockedi _f in a dose-dependent fashion. Block development was slow, with time constants decreasing with drug concentration and ranging from 25.8 s at 10 M to 75.5 s at 1 M UL-FS 49. Block did not develop in cells held at –35 mV, at which voltagei _f channels are closed, indicating that channels must open before blocking occurs. Apparently in contrast with the requirement of negative voltages for block development, block was relieved by hyperpolarization with a time course slower than current kinetics. Due to the hyperpolarization-induced block relief, current/voltage (I/V) relations in the presence of UL-FS 49 displayed inward-going rectification. Experimental data fitted the hypothesis that UL-FS 49 behaves as an open channel blocker of single-ioni _f channels. Block occurs within the pore, at a distance of about 39% of the membrane thickness from its internal side.     

Cardiac Purkinje fibres

Summary The influence of intracellular calcium concentration [Ca²⁺] _i on the steady state membrane currentsi was studied in a range of clamp potentials between –20 and –100 mV. Injection of CaCl₂ or Ca-EGTA (pCa 6) increasedi whereas injection of K-EGTA diminished it. The changes i were attributed to a change in steady state potassium conductance, g_K, by four arguments: i was restricted to potentials negative to –20 mV and depended on clamp potential in an inward rectifying manner. i displayed a reversal potential, E_rev, which followed log [K⁺]₀ with 60 mV for a tenfold change. Since E_rev obtained during Ca injection agreed with E_rev observed during EGTA injection the potassium driving force had to be constant. Wheng _K was blocked by superfusion with 20 mM Cesium neither CaCl₂ nor K-EGTA injection modifiedi .Supported by SFB 38, Membranforschung, project G2     

Genetic characterization of an α-specific gene responsible for sexual agglutinability in Saccharomyces cerevisiae: mapping and gene dose effect

Summary A recessive ag1 mutation leads to specific defect in sexual agglutinability specifically in cells of the yeast Saccharomyces cerevisiae. The cryptopleurine resistance gene cry^R 1, closely linked to the mating type locus, was used to select / strains which emerged from / strains by mitotic nonreciprocal recombination, to genetically analyse ag1, since ag1 is expressed only in mating type. The ag1 gene was found to be linked to the centromere tightly, to met3 at 4.4 cM, and to ilv3 at 12 cM on chromosome X. Sexual agglutinability of cells was shown to be dependent on the dose of the AG1 gene, using / isogenic strains carrying AG1/AG1, AG1/ag1 or ag1/ag1. The sst2-1 mutation did not suppress the ag1 mutation. Based on these results, function of the AG1 gene is discussed.Abbreviations cM centimorgan - FDS first division segregation - NPD nonparental ditype - PD parental ditype - SDS second division segregation - TT tetratype     

The activation of Ca2+-dependent K+ conductance by adrenaline in mouse peritoneal macrophages

Responses to adrenaline in mouse peritoneal macrophages were investigated with perforated and cell-attached patch-clamp recording, and with a combination of the perforated-patch recording and fura-2 fluorescence measurements. Extracellularly applied adrenaline induced a transient outward current (4–10s in duration, 100–500 pA in amplitude) at –40 mV associated with a marked increase in conductance. The adrenaline-induced current [I _o (Adr)] reversed polarity near –80 mV. The reversal potential depended distinctly on the external K⁺ concentration but not on external Cl^– concentration. Removal of external Ca²⁺ did not affect I ^o(Adr) within 2–4 min but subsequent responses to adrenaline were progressively depressed. In contrast, treatment with an intracellular Ca²⁺ chelator, the acetoxymethyl ester of 1,2-bis-(2-aminophenoxy)ethane-N,N,N,N-tetraacetic acid completely abolished I _o(Adr). Furthermore, I _o(Adr) was blocked by bath-applied quinidine and charybdotoxin, but not by tetraethylammonium or apamin. Extracellular application of an ₁-adrenoceptor agonist phenylephrine and of noradrenaline mimicked I _o(Adr). On the other hand, I _o(Adr) was antagonized by a non-selective -adrenoceptor antagonist phentolamine (0.2 M) and an ₁-adrenoceptor antagonist prazosin (0.2 M), but was not affected by an ₂-adrenoceptor antagonist yohimbine (1 M) or a -adrenoceptor antagonist propranolol (1 M). Cell-attached single-channel recordings with the pipette solution containing 145 mM KCl revealed the activation of single-channel currents with a conductance of 40 pS during application of adrenaline outside the patch. Parallel measurements of membrane current and fura-2 fluorescence in the same cell demonstrated a correlation between the rise in [Ca²⁺]_i and an increase in K⁺ conductance. Therefore, it is concluded that adrenaline activates a Ca²⁺-dependent K⁺ conductance by release of Ca²⁺ from internal stores through an activation of an ₁-adrenoceptor.     

Interactions between Neurons in the Amygdala and Hypothalamus during Conditioned Reflex Behavior Involving Choice of Reinforcement Quality in Cats

The organization of distributed amygdalo–hypothalamic and local amygdalar and hypothalamic neural networks was studied in three cats, trained to perform a food–related operant conditioned reflex to light by the active choice of reinforcement quality method; pressing the pedal with a short latent period provided the cats with a bread/meat mixture, while pressing with a long latent period yielded meat. Animals showed sharp differences in their individual abilities to choose one or the other reinforcement. Two cats preferred long–latency pedal–pressing to obtain meat, and were considered to be self–controlled, while the third cat, which preferred short–latency pedal–pressing to obtain the less valuable reinforcement (the bread/meat mixture) was described as impulsive. Chronically implanted semimicroelectrodes were used to record multineuron activity in the basolateral nucleus of the amygdala and the lateral nucleus of the hypothalamus. Interactions between the discharge trains of neighboring and distant neurons were assessed by cross–correlation analysis. Interneuron functional connections were found to predominate significantly in the local neural networks of the basolateral amygdala and distributed amygdalar–hypothalamic networks in the impulsive animal as compared with the self–controlled animals, suggesting a role for these types of connection in forming the individual characteristics of higher nervous activity. The identical incidences of interneuron interactions in the lateral nucleus of the hypothalamus in cats with different individual preferences suggest that local networks in this formation are not involved in analyzing reinforcement quality.