Effect of testosterone loading on the kinetic of faecal testosterone excretion in mallards.

Intestinal passage time of coloured fodder and testosterone turnover were examined by faecal steroid analysis in mallards in the reproductive and postrefractory period. In the latter, the discharge of coloured fodder began 36 minutes after ingestion in males, and 56 minutes in females. During reproduction the discharge began 93 minutes and 112 minutes after ingestion in males and females, respectively. Total passage time was similar in the reproductive and postrefractory period in both sexes. After intraperitoneal testosterone injection, faecal samples were collected for 8 hours and testosterone levels were measured using RIA. In the postrefractory period, 1-2 hours after testosterone loading a strong increase of faecal testosterone content developed in males, meanwhile a slighter testosterone peak appeared in females. During reproduction testosterone excretion began 1.5-2 hours after injection in both sexes but in females its increase was smaller. The duration of response to testosterone loading was 5 hours in both periods and both sexes. Intensive excretion after T loading appeared earlier in males than in females, but total passage time finished at the same time: 5 hours after loading. The character of testosterone excretion was corresponding to the passage of fodder-chimus-faeces in the reproductive and postrefractory period in both sexes.

[Efficiency potential in the pacemaker/implantable cardioverter defibrillator outpatient clinic]. / Effizienzpotential in der Nachsorge von Schrittmachern und implantierbaren Kardioverter Defibrillatoren.

The aim of the present study was to elucidate whether the duration of a technical follow-up (FU) of a pacemaker (PM)/implantable cardioverter defibrillator (ICD) has an impact on cost-effectiveness in the outpatient clinic. We determined the time required for a complete FU of devices from three different manufacturers. In 130 patients (70 VVI/DDD-PM, 60 VVI/DDD-ICD) with either a PM (Phylos, Chorum/Talent, Kappa, EnPulse) or an ICD (Belos, Alto or GEM) the time was recorded for a complete FU including determination of lead impedance, sensing and pacing threshold. The time for activation of individual menue buttons was excluded. On the basis of time required for FU, cost-units (CU) were calculated for 2000 FU/year and for a presumed device longevity (PM 7 years, ICD 5 years). For VVI-PM, the duration of FU was almost identical for devices from different manufacturers (105+/-11 s to 125+/-8 s; p=n.s.). However, analysis of DDD-PM revealed marked differences (140+/-25 s vs 282+/-23 s, p<0.05). Time for FU of ICDs varied between 108+/-5 s and 207+/-21 s (p<0.05) in VVI-ICDs and between 129+/-8 ms and 225+/-23 s (p<0.05) in DDD-ICDs. The total savings could be 55 000 CU in VVI- and 53 333 CU in DDD-ICDs. For full automatic DDD-pacemakers (EnPulse) time for FU could be reduced to 58+/-3 s (p<0.05). Differences in FU times were caused by problems with telemetry, delay during booting of the programmer, interrogation at the beginning and at the end of FU and for sensing tests. Improving not only programmers and devices but also test automaticity could significantly increase cost-efficiency in the outpatient clinic.

T-cell epitope analysis on the autoantigen phogrin (IA-2beta) in the nonobese diabetic mouse.

The protein tyrosine phosphatases (PTPs) IA-2 and phogrin (IA-2beta) are major autoantigens in type 1 diabetes that possess common serological epitopes in their COOH termini. The epitopes recognized by the T-cells that cause the disease, however, remain to be defined. Eight phogrin-specific T-cell clones were generated from NOD mice, and their epitopes were mapped. The mapping was performed initially with recombinant gluthathione S-transferase-phogrin COOH deletion constructs and ultimately with overlapping synthetic peptides. Two dominant epitopes were identified: one (aa 629-649) immediately adjacent to the transmembrane domain (aa 604-628) and the second (aa 755-777) lying in the NH(2)-terminal region of the conserved PTP domain. T-cells that are specific to either of these peptides and that could destroy islet tissue in vivo though spontaneous T-cell proliferative responses were observed in prediabetic female NOD splenocytes only to the aa 755-777 epitope. In NOD female mice immunized with the epitope peptide, intramolecular determinant spreading occurred from the aa 629-649 epitope to the aa 755-777 epitope but not in the opposite direction. We concluded that the initial T-cell response to phogrin is restricted to a small number of dominant peptides and that it subsequently spreads to other regions of the molecule, including those containing the major humoral epitopes that are highly conserved between IA-2 and phogrin.

Cellular immune response to phogrin in the NOD mouse: cloned T-cells cause destruction of islet transplants.

The ability of nonobese diabetic (NOD) mice to mount a cellular immune response to the secretory granule protein tyrosine phosphatase (PTP), phogrin was evaluated by immunization of 8- to 12-week-old animals with recombinant phogrin in complete Freund's adjuvant. Draining lymph nodes displayed a robust proliferative response to the protein, as did derived T-cell lines and clones. Ten clones obtained by limiting dilution were all CD4+ and of a T-helper-1-like phenotype, but showed variation in their Vbeta usage. Of the 10 clones, 3 responded to endogenous antigens in rat islets. Two of these caused the destruction of rat islets that had been transplanted under the kidney capsule of streptozotocin-treated NOD scid mice without affecting adjacent thyroid implants. The results demonstrate the feasibility of generating antigen-specific diabetes-inducing CD4+ cells by direct immunization of NOD mice and their potential use for further studies of the antigenic epitopes in the PTP family members. The conclusion, based on serological studies, that PTP members do not play a role in the pathogenesis of type 1 diabetes in rodent models needs reevaluation in light of these findings.

Differential expression and imprinting status of Ins1 and Ins2 genes in extraembryonic tissues of laboratory mice.

There are two functional insulin genes in the mouse genome. The Ins2 gene is imprinted and expressed monoallelically from the paternal allele in the yolk sac. In the present study we have re-examined the imprinting status of Ins1. We found that Ins1 is not expressed in the yolk sac of several laboratory mouse strains. The asynchrony of replication at the wild type locus was significantly lower than at imprinted loci and was more similar to non-imprinted loci. Finally, we have taken the advantage of the Ins1(neo) allele created by homologous recombination to examine the allelic usage at this locus. We observed that the neo gene inserted at the Ins1 locus was expressed from both the paternally and the maternally transmitted allele. Therefore, the Ins1 gene does not share any of the basic properties of imprinted genes. On the basis of these data, we concluded that Ins1 locus is unlikely to be imprinted in common laboratory mice.

Modification of the cardiac transmembrane potentials and currents by polyamines.

Spermine up to 10(-5) M increased the resting potential (RP) and maximum rate of rise (Vmax) of the action potential (AP) and accelerated the initial (20, 50%) repolarization in isolated guinea-pig and cat atria. These effects were not modified by pindolol (4 X 10(-7) M) or atropine (3 X 10(-7) M) but were prevented by indomethacin (10(-6) M). The right papillary muscle of guinea-pig only showed the acceleration of repolarization while the other parameters were not changed. Spermine did not influence the inward membrane currents under voltage clamp conditions in frog sinoatrial fibers. Spermidine up to 10(-5) M increased RP, Vmax and the duration of AP in guinea pig papillary muscle. RP, Vmax and AP duration were moderately enhanced by spermidine in guinea pig atrium and were not changed in cat atrium. Spermidine was found to stimulate the inward membrane current. Its stimulatory effect was confined to the fast component of the inward current. Neither spermidine nor spermine were able to induce the Ca2+-dependent slow AP in 25 mM K+-depolarized atrium. It is suggested that polyamines influence cardiac membrane events which are responsible for AP and ionic currents.

Antagonism between adenosine and bromobenzoyl-methyladamantylamine, a K+-channel blocker, in atrial myocardium of guinea pig.

The effect of bromobenzoyl-methyladamantylamine (BMA) on the adenosine-induced changes in the electrical and mechanical activity of the atrial muscle, and the effect of adenosine on the slow action potentials induced by BMA in K+-depolarized atrial myocardium of guinea pig were studied. BMA was able to antagonize the adenosine-induced shortening of the action potential duration and negative inotropic effect. This action of BMA was potentiated by theophylline, and reduced by dipyridamole. Adenosine depressed the BMA-induced slow action potentials. The results suggest that there may be an antagonism between BMA and adenosine.

Inhibition of myocardial K+ channels by bromobenzoyl-methyladamantylamine, an adamantane derivative.

The effect of bromobenzoyl-methyladamantylamine (BMA) on transmembrane potentials and contractility of atrial and ventricular myocardium of guinea-pig and cat, as well as on transmembrane ionic currents of the frog atrial trabeculae was studied using conventional glass microelectrode and double sucrose-gap voltage clamp techniques. BMA markedly prolonged the action potential duration, depolarized the cell membrane, reduced the rate of rise of the action potential and exerted a positive inotropic effect on non-clamped myocardial preparations. The drug-induced pacemaker activity in ventricular working muscle of cat. Moreover, BMA antagonized the effects of the K+ channel activator acetylcholine in a dose-dependent manner. BMA was found to induce slow response action potentials in K+ -depolarized ventricular myocardium of guinea-pig. In voltage clamp experiments, BMA reduced the outward K+ current but had no effect on either rapid inward Na+ or slow inward Ca2+ currents. The results suggest that BMA is capable of selectively blocking the myocardial K+ channels.

Blockage of the fast sodium current by dimethindene in frog auricular fibres.

The effect of dimethindene (DMI) on action potential and fast inward Na current (INa) of frog atrial fibres was studied using double sucrose gap voltage-clamp technique. DMI reduced the amplitude and maximum rate of rise of the action potentials without altering the resting membrane potential. The drug inhibited the fast Na conductance in a concentration-dependent manner, without changing the reversal potential. The shape of the current-voltage curve along the voltage axis remained unchanged in the presence of DMI. The time to peak of the INa was not significantly altered by the drug whereas the rate of the INa inactivation (tau h) was slowed. DMI shifted the steady-state inactivation curve of INa (h infinity) to more negative potentials, and increased the reactivation time constant of the sodium system (tau r). The inhibition of INa was use-dependent. The results suggest that DMI interacts with the inactivation mechanism of the sodium channel of the frog myocardium, and explain its favourable antiarrhythmic activity.

(-)Deprenyl and (-)1-phenyl-2-propylaminopentane, [(-)PPAP], act primarily as potent stimulants of action potential-transmitter release coupling in the catecholaminergic neurons.

The activity of the catecholaminergic neurons in the rat brain is enhanced significantly 30 min after the subcutaneous injection of very small doses of (-)deprenyl (threshold doses: 0.01 mg/kg for noradrenergic neurons and 0.025 mg/kg for dopaminergic neurons). As a catecholaminergic activity enhancer (CAE) substance (-)deprenyl is about ten times more potent than its parent compound, (-)methamphetamine. While the (+)methamphetamine is 3-5 times more potent than (-)methamphetammine in releasing catecholamines, the (-)methamphetamine is the more potent CAE substance. The mechanism of the CAE effect of (-)deprenyl and (-)PPAP, a deprenyl-derived substance devoid of MAO inhibitory potency, was studied in rats by measuring: a) the release of catecholamines from striatum, substantia nigra, tuberculum olfactorium and locus coeruleus; b) the stimulation induced release of 3H-noradrenaline from the isolated brain stem; and c) the antagonistic effect against tetrabenazine-induced depression of learning in the shuttle box. The CAE effect was found to be unrelated: a) to the inhibition of MAO activity; b) to the inhibition of presynaptic catecholamine receptors; c) to the inhibition of the uptake of catecholamines; and d) to the release of catecholamines. It was concluded that (-)deprenyl and (-)PPAP act primarily as potent stimulants of action potential-transmitter release coupling in the catecholaminergic neurons of the brain. We show that both (-)deprenyl and (-)PPAP enhance the inward Ca2+ current in sino-auricular fibers of the frog heart. (-)PPAP was much more potent than either (+)PPAP or (-)deprenyl in this test.

The use of an internal mammary artery for creating a systemic-to-pulmonary artery shunt.

We report on two patients who had systemic-to-pulmonary artery shunts created by the use of the internal mammary artery (IMA). The first patients was operated on more then 30 years ago (the case has never been published) and the second one move recently. We give a brief summary of the ten cases of the same operation published so far, and emphasize the usefulness of the IMA that stays open despite the initially poor run-off and is capable of supplying increasing amounts of blood to the growing pulmonary arteries.

A comparative methodical study of the faecal steroid analysis on birds: looking for a valid method of testosterone determination.

In a comparative study, a relatively simple and high sensitivity method was developed for analysis of testosterone-equivalent(s) in the faeces of different bird species. To determine the recovery of extractions and purifications, tritium-labelled testosterone was added to the wet samples. Then the samples were treated with sodium dodecil sulphate (SDS), an emulsificator to "open-up" the complex, lipid-coated particles of faecal samples. This emulsification resulted in the decrease of the quantity of interfering substances after diethyl-ether extraction and the linearity of the measured testosterone equivalents from aliquots in the range of 2 and 10 mg of faeces. In the RIA, we applied a group specific polyclonal testosterone antibody which cross-reacted with reduced metabolites and at a certain level with sulphate conjugates as well. The use of Helix enzymes did not modified significantly the results of the analysis relating to a low level of conjugated androgens in the faecal extracts. The biological validity of the method was tested on domestic cockerels, where between the plasma and faecal testosterone values a four hours phase shift was observed, with a correlation of 0.6355. This method is suitable for "non invasive", behavioural-ethological studies.

[Electrophysiologic study of the anti-arrhythmic mechanism of action of phosphocreatine in acute myocardial ischemia and reperfusion]. / Elektrofiziologicheskoe issledovanie mekhanizmov antiaritmicheskogo deistviia fosfokreatina pri ostroi ishemii i reperfuzii miokarda.

To determine the possible application of exogenous phosphocreatine (ePC) to protect the ischemic myocardium from reperfusion abnormalities in cardiac rhythm, the antiarrhythmic and antifibrillatory activities of the agent were studied in an acute myocardial ischemia model and reperfusion-induced cardiac damage. It was shown that ePC produced a pronounced antifibrillatory effect in acute coronary occlusion and subsequent reperfusion. The agent substantially increased the threshold of electric ventricular fibrillation and the frequency of spontaneous defibrillation. The highest activity was shown by ePC in ischemic myocardial reperfusion. The agent suppressed both rapid inward Na+ current and slow inward Ca2+ current. The effects of ePC on transmembrane ion currents suggest that the agent has a unique electrophysiological mechanism of action involving the properties of Classes I and IV antiarrhythmics, making ePC promising in clinical application in patients with impaired conduction and automatism.

[Electrophysiologic research on the anti-arrhythmia properties of bonnecor--a new dibenzazepine derivative]. / Elektrofiziologicheskoe issledovanie antiaritmicheskikh svoistv bonnekora--novogo proizvodnogo dibenzazepina.

A new dibenzepin derivative, bonnecor, showed marked antiarrhythmic and antifibrillation properties in anesthetized cats. One and 2 mg/kg doses of the drug reduce cardiac assimilation of imposed ventricular electric stimuli, increase ventricular fibrillation threshold with a strong and lasting effect and prevent arrhythmias and fibrillation due to 10-minute occlusion and subsequent reperfusion of the anterior descending branch of the left coronary artery. Experimental holding of atrial trabecular membrane potential in a frog, in conditions of a double saccharose bridge, showed bonnecor, in 5 X 10(-7) and 1 X 10(-6) g/ml concentrations, to inhibit rapid sodium inflow and slow calcium inflow. It is expected that bonnecor can be used clinically to control critical arrhythmias and prevent sudden cardiac death.

[Is the determination of the defibrillation threshold in patients with an implantable cardioverter-defibrillator still required?]. / Wird die Bestimmung der Defibrillationsschwelle bei Patienten mit implantierbarem Kardioverter/Defibrillator noch benötigt?

BACKGROUND: Intraoperative testing of implantable cardioverter-defibrillators (ICDs) is time consuming and associated with risks. In the present study, we elucidated whether the initial implantation of an ICD with high energy output makes intraoperative defibrillation threshold testing (DFTT) unnecessary even though antiarrhythmic (AA) therapy is needed in the future. METHODS: A total of 111 patients (94 men, 17 women) receiving an ICD with subsequent AA therapy (mexiletine, amiodarone, sotalol, flecainide) were analyzed retrospectively. DFT was performed during ICD implantation and after AA drug therapy. In a second step, DFT results from the study cohort were analyzed for implantation of virtual ICDs with either low (≤ 30 J, LOD), intermediate (34 J, IOD), or high energy output (36 J, HOD). RESULTS: In the study cohort, all patients reached the safety margin (SM) of 10 J between DFT and maximal shock energy of the ICD. After loading of AA agents, 6 patients (12%) with a LOD, 3 patients (11%) with an IOD, and 3 (13%) patients with a HOD failed the 10 J SM. Using virtual ICDs, 6 (5.5%) patients with a LOD, 1 patient (1%) with an IOD, and no patients with a HOD would have failed the 10 J SM. After loading of AA agents, 18 patients (16%) with a virtual LOD, 12 patients (10.8%) with an IOD, and still 9 patients (8%) with a HOD would have failed the 10 J SM. CONCLUSION: Our results demonstrate that the 10 J SM would have been achieved intraoperatively in all patients with virtual HOD ICDs. Thus, determination of the DFT during implantation does not seem to be obligatory. However, in patients receiving AA agents, DFT testing is still required.

Indications for predismissal testing with arrhythmia-induction in patients receiving an implantable cardioverter defibrillator.

UNLABELLED: Arrhythmia induction during implantation of cardioverter defibrillators (ICD) is a standard procedure. However, controversy exists regarding the need for routine arrhythmia induction before discharge from hospital (pre-hospital discharge (PHD) test). In order to reduce the number of tests we identified risk factors that predict relevant ICD malfunction. METHODS AND RESULTS: 965 patients receiving a first device implantation (n=724) or device/system replacement (n=241) between 1998 and 2004 were analysed. During implantation 176 (18%) complications (intraoperative undersensing of induced arrhythmias, unsuccessful arrhythmia-therapy or low DFT safety margin) occurred. Frequent (>4 times) intraoperative lead repositioning due to low sensing values was present in 44 patients (5%). 9% of the patients with first ICD implantation, 21% with device replacement and 27% with system replacement developed complications during PHD testing with arrhythmia induction. Intraoperative complications, although corrected during implantation, were independent risk factors for malfunction during PHD testing (p<0.05). Additional predictors for malfunction were intraoperative lead repositioning (>4 times) and a history of both VF and VT (p<0.05). Patients without intraoperative complications rarely developed malfunction during PHD testing (3.7% first device, 6.25% system replacement). Only in patients undergoing device replacement was a higher risk for failure (13%) evident. No risk factors could be identified for these subgroups. CONCLUSION: Routine arrhythmia induction during PHD is recommended in ICD patients with intraoperative complications, although corrected during implantation, as well as frequent intraoperatives lead repositioning. Patients undergoing device/system replacement uncomplicated implantation are not generally at low risk for device failure.