Coronary blood flow in rats native to simulated high altitude and in rats exposed to it later in life

Summary In rats exposed to a simulated high altitude of 3500 m for their whole prenatal and postnatal life a severe cardiac hypertrophy develops. In rats born and first staying 5 weeks at sea level and then being exposed to simulated high altitude, only a unilateral right cardiac hypertrophy occurs. In both groups nutritional coronary blood flow was estimated in left ventricle, right ventricle, and septum and was compared with control animals of similar age. Coronary blood flow was measured at hypoxia in all groups. At first cardiac output was determined by the Fick principle, then⁸⁶Rb was applied and the animals were killed after 55 sec. Activity of⁸⁶Rb was measured in both cardiac ventricles and septum and the fractional uptake was calculated. According to Sapirstein (1956, 1958) the distribution of⁸⁶Rb follows the distribution of cardiac output and from both these data the nutritional blood flow to the parts of the heart may be estimated.Cardiac output was similar in rats exposed to simulated high altitude later in life (newcomers) and in control animals, but it was significantly lower in rats born in the low pressure chamber (natives).Fractions of cardiac output supplying cardiac ventricles and septum in rats from both hypoxic groups were significantly higher than in control animals. In the natives they were significantly higher than in the newcomers. The fractions of cardiac output in both newcomers and natives remained significantly higher than those of the control animals, also when calculated per gram of heart tissue.Nutritional coronary blood flow (in ml/min) was higher in both ventricles and septum of the newcomers and in the right ventricle of the natives, and lower in the septum of the natives, when compared with control animals. Coronary blood flow per gram of heart tissue (in ml/min·g) was significantly higher in all cardiac parts of the newcomers, but it was about the same in all cardiac parts of the natives when compared with controls.The importance of observed changes concerning myocardial tissue oxygenation is analyzed by using Krogh's cylindrical tissue model.Presented in part at the XXVIth International Congress of Physiological Sciences, New Delhi, India, October 20–26, 1974.     

An analysis of postsynaptic potentials of tectal neurons of the frog: correlation with impulses recorded from the terminals of retinotectal afferents

Summary The purpose of this study is to examine the synaptic action between terminals of retinal ganglion cell axons and tectal neurons. To accomplish this, an extracellular single unit identified as retinotectal fiber was first isolated from the superficial layer of the optic tectum and intracellular responses were recorded from a tectal neuron in the vicinity of the extracellular recording electrode. On-off retinal fibers and both E-E (EPSP at on and off of diffuse light) and EI-EI type (EPSP-IPSP combination at on and off of diffuse light) tectal neurons were selected for the pre- and postsynaptic pair. Postsynaptic responses to a small moving square were averaged by triggering with the isolated presynaptic impulses. The latency of the resultant EPSPs indicated that most of the E-E and EI-EI type tectal neurons were monosynaptically activated by on-off retinal fibers. One of the E-E type tectal neurons was identified as a large ganglionic neuron in layer 8.     

Foraging strategies ofDrosophila melanogaster: A chromosomal analysis

Two larval foraging strategies inDrosophila melanogaster were identified, rover and sitter. Rovers traverse a large area while feeding whereas sitters cover a small area. The difference between rovers and sitters was analyzed genetically by chromosomal substitutions between isogenic stocks. Differences in larval locomotor behavior (crawling behavior) can be attributed to the second chromosome, the rover strategy being dominant over the sitter strategy. Differences in feeding rate (shoveling behavior) are affected additively by both the second and third chromosomes. Natural populations ofDrosophila larvae were sampled three times over a 2-month period; rovers and sitters were at constant frequencies in these populations. The two foraging strategies are discussed in the light of resource utilization in environments where food is distributed continuously or discontinuously.     

Prostate specific antigen and prostate specific acid phosphatase in adenocarcinoma of Skene's paraurethral glands and ducts

An autopsy case of adenocarcinoma of Skene's paraurethral gland co-incident with renal cell carcinoma is described. The adenocarcinoma showed distinct prostate specific antigen and prostate specific acid phosphatase pointing to the equivalence between the male prostate and Skene's paraurethral glands and ducts. Skene's gland are the homologue of the prostate in females and tumours arising from them are immunohistochemically similar to male prostate carcinoma.In the title and text the authors used the official term of Nomina Anatomica paraurethral (Skene's) glands and ducts. Nevertheless recently published data on cross-antigenicity between the male prostate and Skene's glands and the newly discovered exocrine and neuroendocrine parameters of the prostate homologue in the female, comparable with the male prostate (Zaviai 1987), support the use of the same term — the prostate — for prostatic tissue in both sexes (Zaviai 1987, Zaviai et al. 1985). The designations female prostate homologue or female prostate equivalent are a compromise between terms the female prostate and Skene's paraurethral glands.     

Gravity responses of Purkinje cells in the nodulus

Summary In cats, either decerebrated or under chloralose anaesthesia, Purkinje cells (P-cells) of the cerebellar nodulus have been examined with the animal under static lateral tilt (roll±20°). The cell activity was extracellularly recorded and both simple and complex spike discharge patterns were studied.In 20 cells out of a population of 198, simple spike firing was found to be affected by static roll. Ten cells had an -type response, 8 a -type, while only single examples of and activations were found.Out of 67 Purkinje cells tested for complex spike activation, 5 were found to be sensitive to static roll, 4 with an or response and one with a response.The results are to be attributed to pure otolith activation and show that this input is able to modulate P-cell activity in the nodulus through both the mossy fibre and the climbing fibre systems.     

Adaptation and habituation of the vestibulo-ocular reflex in intact and inferior olive-lesioned rats

Summary The gain of the vestibulo-ocular reflex (VOR) of intact pigmented rats was adaptively modified by training protocols that created a visual-vestibular conflict. For training, head restrained animals were oscillated on a turntable in front of an optokinetic pattern projected onto a cylindrical wall. The optokinetic pattern either moved the same amplitude with the animal (in-phase: 0.05 Hz ± 20°/s) or opposite in direction (out-of-phase: turntable and pattern 0.05 Hz ± 10°/s each). VOR responses were tested in darkness before and after each 8 min training period for a duration of 40 min. During out-of-phase training the gain of compensatory eye movements measured in light was close to 2 from the beginning on and the VOR tested in darkness increased in gain progressively from 0.48 (±0.12) to 0.9 (±0.3; P < 0.05) in 5 out of 7 rats. Two rats did not adapt their VOR gain. Phase values decreased slightly by about 10°. During in-phase stimulation compensatory eye movements were almost completely suppressed (gain close to 0) from the beginning on and the VOR tested in darkness decreased gradually in gain from 0.62 (±0.17) to 0.13 (±0.1; P<0.001) in all 6 trained rats. Phase values decreased in parallel from 151° to 119° (P< 0.01). The effectiveness of the in-phase training paradigm in the absence of compensatory eye movements indicates that retinal image slip is the relevant signal for adaptation. In seven rats with histologically verified almost complete inferior olive (IO) lesions (chemically induced at least 45 days prior to training), out-of-phase and in-phase stimulation evoked compensatory eye movements with gains comparable to those in intact rats. VOR parameters measured in darkness were altered with respect to those of control rats. Gain differed extremely between individuals and phase lag re acceleration was in all IO-lesioned rats larger than in intact rats. The time constant of the VOR in response to table velocity steps was significantly longer (17 s ±4) than in intact rats (11 s ± 3). Training did not alter the gain of the VOR in 5 out of 7 IO-lesioned rats. One rat increased its gain during out-of-phase training in the first, but not during a second training session (and not during in-phase training) and another rat decreased its gain during in-phase training (but not during out-of-phase training). These changes in VOR gain might have occurred by chance rather than by learning. The absence of adaptation in IO-lesioned rats can be explained either by the absence of climbing fiber mediated slip signals in the cerebellar cortex or by lesion-induced secondary changes which result in a long-term reduction of the inhibitory efficacy of Purkinje-cells. In the absence of arousing stimuli VOR responses of intact rats exhibit a strong decrement during table oscillations in darkness. Between trials, with the rat at rest, response magnitude recovered spontaneously. Six out of 8 IO-lesioned rats expressed a very similar modification of their VOR gain. These results indicate that the neural mechanisms responsible for adaptive gain decrease during in-phase training and those responsible for a gain decrease during short-term habituation are different.     

MHC Class I–Subversive Gene Functions of Cytomegalovirus and their Regulation by Interferons–an Intricate Balance

Multiple glycoproteins of human cytomegalovirus (HCMV) encoded by the genes US2, US3, US6 and US11 interrupt the MHC class I pathway of antigen presentation at distinct checkpoints to avoid recognition of infected cells by cytotoxic CD8+ T lymphocytes. The action of cytokines like interferon (IFN)-, IFN-/ and tumour necrosis factor (TNF-) compensate for the viral inhibition and restore antigen presentation in HCMV-infected cells. This finding was explained by their effects on cellular rather than viral genes and reflected by an increase in the production, assembly and maturation of MHC class I molecules resulting in an escape of MHC I from viral control. Here we reproduce the IFN--mediated effect when MHC I-subversive gene functions of HCMV are tested in isolation, but the efficacy of IFN- to restore MHC I surface expression in US2-, US6- and US11-transfectants differs significantly. In addition, in HCMV-infected cells IFN- strongly affects the synthesis of the US6-encoded glycoprotein. Despite the capability of HCMV to block the interferon signaling pathway the IFN- driven enhancement of MHC class I and class II expression remains effective provided that cells are exposed to IFN- before infection. Our findings illustrate a complex interplay between host immune factors and viral immune evasion functions.     

Facilitation by 5-hydroxytryptamine of ATP-activated current in rat pheochromocytoma cells

The effects of 5-hydroxytryptamine (5-HT) on an inward current activated by extracellular ATP were investigated in rat pheochromocytoma PC12 cells. Under whole-cell voltage-clamp conditions 5-HT (10 M) reversibly enhanced the amplitude of the current activated by 30 M ATP. The enhancement may not be due to an increase in the number of functional channels because the current activated by 300 M ATP was not remarkably augmented compared with the current activated by 30 M ATP. The current enhancement by 100 M 5-HT was less obvious than that by 10 M 5-HT. When the current kinetics were compared, activation of the ATP-evoked current was accelerated to the same extent by either 10 or 100 M 5-HT, whereas deactivation was largely more accelerated by 100 M 5-HT. Propranolol (10 M), a 5-HT₁ receptor antagonist, or LY53857 (10 M), a 5-HT₂ receptor antagonist, exerted an agonistic effect: the ATP-activated current was facilitated by these compounds. Metoclopramide (10 M), a 5-HT₃ receptor antagonist, neither facilitated the ATP-activated current, nor blocked the current facilitation by 5-HT. Guanosine 5-O-(2-thiodiphosphate) (GDP[S]) (2 mM), the non-hydrolysable analog of guanosine 5-triphosphate (GTP), or K-252a (2 M), a protein kinase inhibitor, did not affect the facilitation by 5-HT of the ATP-activated current when they were included in the intracellular solution. The ATP-activated current pre-facilitated by 10 M dopamine was not enhanced by 10 M 5-HT. Similarly, the pre-facilitation by 5-HT attenuated the current enhancement by dopamine. The results suggest that 5-HT facilitates the ATP-activated channels through receptors that are not readily classified into conventional subclasses of 5-HT receptors. The reciprocal masking between the current facilitation by 5-HT and that by dopamine, combined with their sensitivities to the compounds involved in the intracellular solution, indicates that the facilitation by 5-HT may share not all, but some, common cellular mechanism with that by dopamine.     

Further observations on the morphogenesis of human rotavirus in MA 104 cells

Summary Human rotavirus KUN strain was cultivated in a fetal rhesus monkey kidney cell line, MA 104 cells. Four types of virus particles in cells infected with KUN strain were clearly identified: nucleoid cores, single-shelled particles, double-shelled particles, and membrane band, enveloped particles. Enveloped particles were found only in the thin sections of infected cells. When first visible, the virus precursors appeared at the ribosome free membrane of rough endoplasmic reticulum (RER), increasing in size while simultaneously being coated with nucleocapsid, inner shell. Single-shelled particles were also synthesized within bundles of filaments of viroplasm in the cytoplasma. During subsequent virus maturation two types of budding processes were observed. Double-shelled particles arising at the RER membrane entered the cisternae of the RER through an exocytosis-like process. In contrast, the enveloped particles developed in the cisternae by being completely enclosed with RER membrane, and later during cytolysis released the single-shelled particles. These enveloped virus particles appeared to be the result of inefficient virus maturation at the last stage of outer capsid formation.With 11 Figures     

Is the voltage divider ratio a reliable estimate of the resistance ratio of the cell membranes in tubular epithelia?

The length dependence of the voltage divider ratio (VDR) was investigated in a double cable model of tubular epithelia with point source current injection into the tubular lumen in order to find out, whether there is a region, in which the VDR — as in flat sheet epithelia — is an appropriate measure of the relative magnitude of the apical (r _a) and basal (r _b) cell membrane resistances. Irrespective of the choice of the cable parameters, we find that VDR, defined as luminal over cellular voltage deflection, overestimates the resistance ratio (r _a+r _b):r _b near the origin, but underestimates it at distances () greater than 1 luminal length constant (). In the region < there is a crossover point, where VDR is an accurate estimate of the resistance ratio. If the difference between VDR at the origin and at large distances (>) is small, then VDR is a good estimate of the resistance ratio. This is also true, if VDR is constant between 0.5 and >, (with the exception of some cases, in which the longitudinal resistance in the cell column is exceedingly high). If the latter conditions do not apply, we find that VDR, as measured at =, underestimates the resistance ratio at worst only by 8.8%, provided the cable properties are such that the luminal voltage attenuation exhibits only one single exponential (with maximum tolerable amplitude deviation of 5% at the origin). Cable analysis measurements on rat proximal tubule indicate that VDR is constant in the range between 0.5 and >. Hence (VDR)₌ may be considered as a valid approximation of the ratio of cell membrane resistances in this epithelium.Main symbols and notations r _a resistance per unit length of luminal membrane ( cm) - r _b resistance per unit length of basal membrane ( cm) - r _s resistance per unit length of shunt pathway ( cm) - r _i core resistance of luminal cable per unit length (/cm) - r _c core resistance of cellular cable per unit length (/cm) - distance from current source (cm) - V _i the change in the lumen potential due to current injection (V) - V _c the change in the cellular potential due to current injection (V) - I _o injected current (A) - _i length constant of the luminal cable if isolated from the cellular cable (cm) - _c length constant of the cellular cable if isolated from the luminal cable (cm) - , length constants actually observed in the double cable (cm) - i _a current flow through the apical resistance at (A/cm) - i _b current flow through the basal resistance at (A/cm) - i _s current flow through the shunt pathway at (A/cm) - i _c current flow along the cellular cable at (A) - i _i current flow along the luminal cable at (A) This work was supported by the Deutsche ForschungsgemeinschaftDedicated to Prof. T. Hoshi, Dept. of Physiology, University of Tokyo, Japan, a pioneer in renal tubular resistance analysis     

Präzisierung der Reizwirkung mittelfrequenter Wechselströme

Zusammenfassung Bei der Mittelfrequenz-Impuls-Reizung ist streng darauf zu achten, daß keine polaritären Reizkomponenten auftreten. Die diesbezügliche Kontrolle wird am besten mit Hilfe des Konvertibilitätstestes vorgenommen, d. h., es darf beim Vertauschen der Zuführungen zu den Reizelektroden weder die Reizschwelle bzw. die Größe des kollektiven Reizerfolges noch dessen Latenzzeit eine signifikante Änderung erfahren. Auf diese Weise wird die Phasenunabhängigkeit des echten Mittelfrequenz-Reizeffektes nachgewiesen.Diesen Anforderungen entsprechen Mittelfrequenz-Impulse, deren Trägerfrequenz über einige wenige Perioden sich aufschaukelt und ebenso wieder abklingt. Demgegenüber sind Mittelfrequenz-Stromstöße mit phasenstarrem Einsatz und Ende nicht unbedingt frei von polaritären Ein- bzw. Ausschalt-effekten, indem sowohl die erste als auch die letzte Trägerperiode einen polaritären Wechselimpuls-Reizeffekt ergeben kann, je nach Phasenlage bezogen auf die wirksame Reizelektrode und Art der Ansprechbarkeit des Reizobjektes (Nerv) auf entsprechend kurze gleitspiegelsymmetrische Wechselimpulse. Für eine echte Mittelfrequenz-Stromstoß-Reizung ist demnach ebenfalls ein Aufschaukeln und Abklingen der Trägerfrequenz über einige wenige Perioden erforderlich.Es besteht ein prinzipieller Unterschied zwischen der echten Mittelfrequenz-Reizung, die phasen -bzw. periodenunabhängig ist und schon früher als apolaritär bezeichnet wurde, und der konventionellen polaritären Reizung, die als polaritäre Komplikation der Mittelfrequenz-Reizung auftreten kann.Diese Präzisierung der Reizwirkung mittelfrequenter Wechselströme wurde angeregt durch zwei im Text erwähnte Publikationen, in denen in keineswegs überzeugender Weise versucht wird, die Mittelfrequenz-Reizung letzten Endes auf das polare Gesetz der Erregung zurückzuführen.

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Summary The particular excitatory action exerted by middle-frequency alternating current can only be revealed if care is taken to eliminate the occurrence of so-called polarity effects. Such effects are produced by the short alternating impulses represented by the first and the last period of a middle-frequency current pulse and are based on the polar law of excitation.In order to prevent such polarity intrusions, it is necessary to increase and decrease the amplitude of the middle-frequency current pulses over a few carrierperiods, or, to use amplitude-modulated middle-frequency impulses of variable shape and duration of envelope.A true middle-frequency excitatory effect is easily demonstrated by resorting to the convertibility test. It will then become evident that stimulation threshold, magnitude as well as latency of response do not change during reversal of the stimulating poles. This means, that no significant phase change of the response with regard to the carrier-frequency occurs when the leads to the stimulating electrodes are commuted, and that, as a result, true middle-frequency effects do not depend upon one particular catelectrotonic variation among the carrier-periods of a middle-frequency current pulse.It can thus be concluded that a fundamental difference exists between true middle-frequency stimulation, which is based on a non-polarity or apolarity principle, and the conventional stimulation of the polar or polarity type.This paper has been written in the hope of dispelling some errors of interpretation (discussed in the text) tending to ascribe the excitatory effects of middle-frequency impulse stimulation to the classical polar law of excitation.

     

Molecular organisation of an A mating type factor of the basidiomycete fungus Coprinus cinereus

Summary The A3 and A3 genes, which together constitute the A42 mating type factor of Coprinus cinereus, were isolated from a cosmid genomic library by walking 50 kb, a map distance of 0.5 units, from the closely linked metabolic gene pab-1. Cosmid clones having A gene function were identified by transformation into compatible A6 (22) and A5 (11) host cells where either 3 or 3 was expected to elicit the A factor — regulated development of unfused clamp cells. DNAs were digested with various enzymes before transformation in order to identify the smallest fragments containing an active 3 or 3 gene. Two non-overlapping fragments were identified as containing the 3 and 3 genes respectively. Southern hybridisation analyses showed that these two cloned genes had no detectable sequence homology, and that there was little or no hybridisation to the and gene alleles that constitute the A5 and A6 factors. 3 and 3 were shown to be less than 2.0 kb apart and embedded in a DNA sequence extending over 9.0 kb which was unique to our A42 strain and may contain a third A factor gene.     

Arachidonic acid inhibits hormone-stimulated cAMP accumulation in the medullary thick ascending limb of the rat kidney by a mechanism sensitive to pertussis toxin

The possible regulation of adenosine 3,5-cyclic monophosphate (cAMP) accumulation by arachidonic acid (AA) was studied in segments, microdissected from the rat kidney, which are sensitive to arginine vasopressin (AVP). In the presence of 5 M indomethacin, the addition of 5 M AA did not impair AVP-dependent cAMP accumulation (measured during 4 min at 35° C) in the cortical or outer medullary collecting tubule, but decreased this response in the thick ascending limb with an inhibition much more pronounced in the medullary portion (MTAL) than in the cortical portion. In MTAL, the response to 10 nM AVP was inhibited by 34.4±9.6% (SEM) and 65.8±5.4% with 1 M and 5 M AA, respectively, N=5 experiments. AVP-, glucagon- and calcitonin-sensitive cAMP levels in MTAL were inhibited by 5 M AA to a similar extent. AA-induced inhibition was unaffected by the presence of inhibitors of AA metabolism: (1) either 10 M indomethacin or 50 M ibuprofen added to all media; (2) a 10-min pre-incubation and a 4-min incubation of MTAL samples with 10 M eicosa-5,8,11,14-tetrayonic acid, (3) a 1-h preincubation with either 30 M SKF-525A, 20 M ketoconazole, or 20 M nordihydroguariaretic acid. In contrast to AA, 11 other saturated or unsaturated fatty acids had no inhibitory effect on the AVP-dependent cAMP level. In fura-2-loaded MTAL samples, AA induced a slow increase of the intracellular calcium concentration ([Ca²⁺]_i) which reached 21.0±3.8 nM and 92.9 ±21.4 nM over basal values (n=11) at 2 min and 4 min, respectively, after the beginning of the superfusion of 5 M AA. AA-induced inhibition of AVP-dependent cAMP accumulation was due neither to the increase in [Ca²⁺]_i elicited by AA, nor to an activation of protein kinase C because this inhibition: (1) was not blocked when MTAL samples were incubated either in zero Ca²⁺ medium, or in the presence of 1,2-bis(2-aminophenoxy)ethane-N, N, N N-tetraacetic acid (BAPTA) to chelate [Ca²⁺ _i, and (2) it was not reproduced by a pre-treatment of MTAL segments with a phorbol ester. Pre-incubation of MTAL (6 h at 35° C) with 500 ng/ml pertussis toxin (PTX) prevented AA-induced inhibition: in the presence of PTX inhibition was 24.7±6.6% vs 10 nM AVP, as compared to 81.6 ± 4.0% in control groups, i.e in the absence of PTX, N=6. AA had no effect on the cAMP level induced by 5M forskolin. It is concluded that AA inhibits AVP-dependent cAMP accumulation in the rat MTAL by a mechanism which implicates a GTP-dependent protein sensitive to PTX.     

Zur flammenphotometrischen Calciumbestimmung im Urin

Zusammenfassung Für die Calciumbestimmung im Urin wird eine einfache und rasche flammenphotometrische Methode ohne vorherige Fällung des Calciums beschrieben. Der Kationenfehler kann dabei durch das Korrekturverfahren weitgehend ausgeschaltet werden, der Phosphat- und Sulfatfehler durch das Parallelverfahren, während der Bicarbonatfehler infolge Ansäuerung entfällt.Der mittlere Gesamtfehler der Methode beträgt etwa ± 7% (± 2).     

Causal dimensions of college students' perceptions of physical symptoms

According to attribution theory, controllability, locus, and stability are important dimensions underlying causal explanations. The extent to which these theoretical dimensions underlie lay explanations for physical symptoms is unclear. Accordingly, in this study, attributes relevant to the lay public were empirically derived using a multidimensional scaling (MDS) procedure. Undergraduates (N=194) provided similarity judgments for 18 potential causes of physical discomfort. The MDS analysis yielded a three-dimensional solution. The first dimension captured the distinction between physical and nonphysical causes. The second dimension distinguished either variable versus stable causes or those that are controllable versus uncontrollable by health care professionals. The third dimension differentiated causes under low versus high personal control. These findings empirically confirm the theoretically proposed dimensions of personal control and stability and suggest the utility of considering the physical/nonphysical and controllability by health care professional distinctions in future work on attributions in the health domain.     

Cytokine mRNA in the Joints and Draining Lymph Nodes of Rats with Adjuvant Arthritis and Effects of Cyclosporin A

TNF- and IL-1 promote leukocyte recruitment to arthritic joints and may contribute to cartilage degradation while regulatory cytokines such as IL-4 and IL-1RA may in part determine the course of arthritis. Here we report the pattern of TNF-, IL-1, IL-6, IFN-, IL-1RA, and IL-4 mRNA expression, detected by RT/PCR, in the talar joint and draining popliteal lymph node (PLN) of rats with adjuvant arthritis (AA). Levels of TNF- and IFN- mRNA were increased in the PLN before clinical signs of arthritis. This was followed by increases in IL-1 and IL-1RA mRNA at d9 and IL-6 mRNA at d12. PLN IL-1RA mRNA levels were positively correlated with those of IL-1 and TNF- throughout d5-d20. IL-4 mRNA levels were highest on days 7 and 20. In the synovium, a small increase in TNF-, IL-1, and IL-6 mRNA was detected on d5 then again on d12. Maximal synovial TNF- levels were reached on d20, while IL-1 peak expression was on d16 and IL-6 on d14. IL-4, IL-1RA, and IFN- mRNA was undetectable in the synovium. Cyclosporin treatment for 4 days, initiated at the height of arthritis, rapidly decreased clinical disease, and decreased migration of neutrophils and T lymphocytes into the joints. Yet no significant effect of CyA was observed on inflammatory cytokine expression, although the correlation between PLN IL-1RA and IL-1 or TNF- was lost in treated animals. Thus there is a variable pattern of cytokine gene expression in rat AA, the undetectable IL-4 and IFN- mRNA in synovium being analogous to human rheumatoid arthritis.     

Three-dimensional reconstruction of transmitter-identified central neurons by “en bloc” immunofluorescence histochemistry and confocal scanning microscopy

Summary A new method for three-dimensional reconstruction of transmitter-identified neurons is presented which involves en bloc immunofluorescence histochemistry and confocal scanning microscopy. The technique was applied to different types of neurons in the rat brain and lamprey spinal cord. Thick sections or tissue blocs (50–200 m thick) were incubated with antisera against neuropeptides or monoaminergic markers, followed by fluorescent secondary antibodies. Three-dimensional reconstructions were obtained by scanning the preparations in sequential focal planes with a thin laser beam, while sampling the emitted light in each focal plane. The method is convenient and can be applied to a wide variety of neuron types. The reconstructions obtained are accurate since the optical serial sections of the specimen are perfectly aligned, and optic disturbances such as halo phenomena do not occur.     

Mechanisms underlying facilitation by dopamine of ATP-activated currents in rat pheochromocytoma cells

Mechanisms underlying facilitation by dopamine of extracellular adenosine 5-triphosphate (ATP)-activated current were investigated in rat pheochromocytoma PC12 cells using the whole-cell voltage-clamp techniques. Dopamine (10 and 100 M) augmented the peak amplitude of an inward current elicited by ATP (3–100 M). The activation time course of the ATP-evoked current was accelerated by dopamine; the presence of 10 M dopamine shifted the dependence of activation rate constants on the concentration of ATP toward a lower concentration range two fold. Dopamine also accelerated the inactivation and the deactivation, which was determined from the current decay upon washout of ATP. Intracellular mediators responsible for the dopamine-induced facilitation was estimated by loading various compounds in patch pipettes. Facilitation was not observed when K-252a (1 M), a protein kinase inhibitor, was included in the intracellular solution. In addition, facilitation was also attenuated by intracellular adenosine 5-O-(thiotriphosphate)tetralithium salt (ATPS (1 mM) or --methylene ATP (1 mM). Inclusion of adenosine 3, 5-cyclic monophosphate sodium salt (cAMP, 100 M), guanosine 3,5-cyclic monophosphate sodium salt (cGMP, 100 M), 12-O-tetradecanoylphorbol-13-acetate (TPA, 1 M) or phorbol-12,13-dibutylate (1 M) in the intracellular solution did not affect the facilitation. Guanonsine 5-O-(thiotriphosphate)tetralithium salt (GTPS, 500 M) or guanosine 5-O(2-thiodiphosphate)-trilithium salt (GDPS, 500 M) did not modify the facilitation either. The results suggest that dopamine augments the ATP-activated inward current by facilitating association of ATP to its binding site, and that the augmentation may be mediated through some protein kinase which is different from cyclic-nucleotide-dependent protein kinases or protein kinase C.     

Immunologische Untersuchungen an heterogen zusammengesetzten Plasmocytom-γ-Globulinen

Zusammenfassung Durch Kombination der Stärkegel-Elektrophorese mit immunologischen Techniken (Stärkegel-Immunoelektrophorese und Doppeldiffusionsversuche) ist es möglich, das -Globulin des Normalserums und des Plasmocytoms näher zu differenzieren. Zunächst kann festgestellt werden, daß das -Globulin anodenwärts bis in den Bereich der Haptoglobine wandert. Kleinmolekulare -Globulinkomponenten wie das Bence-Jones-Protein können die Beweglichkeit des Transferrins besitzen und hier durch Präcipitation in der Stärkegel-Immunoelektrophorese nachgewiesen werden.Die einzelnen Subfraktionen des Plasmocytom--Globulins sind mit dem normalen -Globulin antigenverwandt, aber nicht identisch. In den Doppeldiffusionsversuchen, die sich in der Kombination Stärkegel/Agargel durchführen lassen, zeigt sich, daß auch die Paraproteinkomponenten, deren elektrophoretische Beweglichkeit von der des normalen 7S -Globulin abweicht, mit Immunserum gegen -Globulin präcipitiert werden. Unter den Präcipitaten lassen sich die des normalen und des abnormen -Globulins unterscheiden. Mit dem hier verwandten Anti--Globulinserum reagierten die Paraprotein-Subfraktionen gleichartig; es ist aber möglich, daß sich bei Verwendung individualspezifischer Immunseren oder Immunseren gegen einzelne Subfraktionen Unterschiede der Antigenstruktur fassen lassen.II. Teil der 1962 mit dem Theodor-Frerichs-Preis der Deutschen Gesellschaft für Innere Medizin ausgezeichneten Untersuchungen. I. Teil: Z. Naturforschg.17b, 598 (1962)II. Teil der 1962 mit dem Theodor-Frerichs-Preis der Deutschen Gesellschaft für Innere Medizin ausgezeichneten Untersuchungen. I. Teil: Z. Naturforschg.17b, 598 (1962).