Role of prothrombin 19911 A>G polymorphism,blood group and male gender in patients with venous thromboembolism: Results of a German cohort study

Journal of Thrombosis and Thrombolysis - The role of the A>G polymorphism at position 19911 in the prothrombin gene (factor [F] 2 at rs3136516) as a risk factor for venous thromboembolism...     

Dopamine exerts no acute effects on Kv1.3 in activated encephalitogenic T cells

Apart from a central function in the extrapyramidal motor system, dopamine has been suggested to play a role in neuroimmune interactions. Particularly in diseases of the central nervous system, such as multiple sclerosis, alterations in dopamine homeostasis might have immunological consequences. We investigated potential effects of dopamine stabilized by ascorbic acid on specifically activated encephalitogenic T cells at the peak of activation. Those cells exhibited an upregulation of voltage-sensitive K+ channels which play a role in many neurotransmitter responses of lymphocytes and fulfilled a prerequisite to respond to dopamine, i.e. stable expression of mRNA for dopamine receptors DRD1, DRD2 and DRD3. However, whole-cell and perforated whole-cell recordings revealed no change in voltage-sensitive K+ currents. Moreover, T cell proliferation was not changed in the presence of dopamine. Previously reported dopamine effects on T cells may be explained by a comparatively lower activation of the cells under investigation, suggesting an activation dependence of dopamine effects that may not be mediated by K+ channels. Alternatively, the occurrence of dopamine degradation products under unprotected conditions may account for the changes reported. Nevertheless, care should be taken when using the dopamine-protecting anti-oxidant ascorbic acid, since we found that it markedly inhibited both K+ currents and lymphocyte proliferation at higher concentrations.     

An impaired neocortical Ih is associated with enhanced excitability and absence epilepsy

Neuronal subthreshold excitability and firing behaviour are markedly influenced by the activation and deactivation of the somato-dendritic hyperpolarization-activated cation current (Ih). Here, we evaluated possible contributions of Ih to hyperexcitability in an animal model of absence seizures (WAG/Rij rats). We investigated pyramidal neurons of the somatosensory neocortex, the site of generation of spike-wave discharges. Ih-mediated functions in neurons from WAG/Rij rats, Wistar rats (sharing the same genetic background with WAG/Rij, but less epilepsy-prone) and ACI rats (an inbred strain, virtually free of seizures) were compared. We complemented whole-cell recordings from layer 2-3 pyramidal neurons with immunohistochemistry, Western blot and RT-PCR analysis of the h-channel subunits HCN1-4. The fast component of Ih activation in WAG/Rij neurons was significantly reduced (50% reduction in the h-current density) and four times slower than in neurons from nonepileptic Wistar or ACI rats. The results showing decreases in currents corresponded to a 34% reduction in HCN1 protein in the WAG/Rij compared to the Wistar neocortex, but HCN1 mRNA showed stable expression. The other three Ih subunit mRNAs and proteins (HCN2-4) were not affected. The alterations in Ih magnitude and kinetics of gating in WAG/Rij neurons may contribute to augmented excitatory postsynaptic potentials, the increase in their temporal summation and the facilitation of burst firing of these neurons because each of these effects could be mimicked by the selective Ih antagonist ZD 7288. We suggest that the deficit in Ih-mediated functions may contribute to the development and onset of spontaneously occurring hyperexcitability in a rat model of absence seizures.     

High K+-induced contraction requires depolarization-induced Ca2+ release from internal stores in rat gut smooth muscle

Aim:

Depolarization-induced contraction of smooth muscle is thought to be mediated by Ca²⁺ influx through voltage-gated L-type Ca²⁺ channels. We describe a novel contraction mechanism that is independent of Ca²⁺ entry.

Methods:

Pharmacological experiments were carried out on isolated rat gut longitudinal smooth muscle preparations, measuring isometric contraction strength upon high K⁺-induced depolarization.

Results:

Treatment with verapamil, which presumably leads to a conformational change in the channel, completely abolished K⁺-induced contraction, while residual contraction still occurred when Ca²⁺ entry was blocked with Cd²⁺. These results were further confirmed by measuring intracellular Ca²⁺ transients using Fura-2. Co-application of Cd²⁺ and the ryanodine receptor blocker DHBP further reduced contraction, albeit incompletely. Additional blockage of either phospholipase C (U 73122) or inositol 1,4,5-trisphophate (IP₃) receptors (2-APB) abolished most contractions, while sole application of these blockers and Cd²⁺ (without parallel ryanodine receptor manipulation) also resulted in incomplete contraction block.

Conclusion:

We conclude that there are parallel mechanisms of depolarization-induced smooth muscle contraction via (a) Ca²⁺ entry and (b) Ca²⁺ entry-independent, depolarization-induced Ca²⁺-release through ryanodine receptors and IP₃, with the latter being dependent on phospholipase C activation.     

Narkosetiefe bei Intubation

In order to study the depth of anaesthesia during endotracheal intubation, 30 patients received either thiopentone or propofol for anaesthesia induction. The BIS value as a parameter for the depth of anaesthesia and the rate pressure product (RPP) were aquired online. Patients who received thiopentone for anaesthesia induction showed significantly higher BIS values at the moment of intubation and reached BIS values >60 significantly more frequently than patients receiving propofol. The RPP in the propofol group lay significantly below that of the thiopentone patients. For all patients there was an mean increase in BIS values of 8 index points and an increase in the RPP. Therefore, BIS values around 50 should be achieved before intubation in order to avoid the critical BIS value for awareness of >60 despite the increase caused by the intubation procedure. Within 24 h of intubation all patients were interviewed for possible signs of awareness. None of the patients was able to remember the intubation or reported other experiences that indicated an unconscious awareness. Nevertheless, the progress of BIS values in a standardized intubation as performed in the normal clinical routine, shows that the use of thiopentone for initiating anaesthesia results in a very flat level of anaesthesia during intubation. The risk for patients to experience awareness should therefore, not be underestimated. Therefore, when using thiopentone it is recommended to also use a rapid acting muscle relaxant or to select a high ED95 to compensate for the flat level of anaesthesia. Alternatively, repetetive boluses of the hypnotic shortly before intubation should be considered or to revert to propofol. The dosage and pharmacokinetics of the analgesic should also be taken into consideration because an insufficient analgesia leads to a faster flattening of the depth of anaesthesia.     

Clinical evaluation of a simultaneous closed-loop anaesthesia control system for depth of anaesthesia and neuromuscular blockade*

We developed a closed-loop system to control the depth of anaesthesia and neuromuscular blockade using the bispectral index and the electromyogram simultaneously and evaluated the clinical performance of this combined system for general anaesthesia. Twenty-two adult patients were included in this study. Anaesthesia was induced by a continuous infusion of remifentanil at 0.4 μg.kg(-1) .min(-1) (induction dose) and then 0.25 μg.kg(-1) .min(-1) (maintenance dose) and propofol at 2 mg.kg(-1) 3 min later. The combined automatic control was started 2 min after tracheal intubation. The depth of anaesthesia was recorded using bispectral index monitoring using a target value of 40. The target value of neuromuscular blockade, using mivacurium, was a T1/T1(0) twitch height of 10%. The precision of the system was calculated using internationally defined performance parameters. Twenty patients were included in the data analysis. The mean (SD) duration of simultaneous control was 129 (69) min. No human intervention was necessary during the computer-controlled administration of propofol and mivacurium. All patients assessed the quality of anaesthesia as 'good' to 'very good'; there were no episodes of awareness. The mean (SD) median performance error, median absolute performance error and wobble for the control of depth of anaesthesia and for neuromuscular blockade were -0.31 (1.78), 6.76 (3.45), 6.32 (2.93) and -0.38 (1.68), 3.75 (4.83), 3.63 (4.69), respectively. The simultaneous closed-loop system using propofol and mivacurium was able to maintain the target values with a high level of precision in a clinical setting.     

Onset properties of mivacurium measured by mechanomyography depend on the twitch height of the adductor pollicis muscle

INTRODUCTION: The influence of the twitch height of the adductor pollicis muscle during baseline measurements on the pharmacodynamic parameters of mivacurium was prospectively evaluated. PATIENTS AND METHODS: Fifty adult patients were anaesthetized with propofol and alfentanil. Neuromuscular function was monitored mechanomyographically by measuring the force of the adductor pollicis muscle following stimulation of the ulnar nerve. Following a stabilization period of 20 min, the individual twitch height of the adductor pollicis muscle was determined before a single bolus of mivacurium (75 microg kg-1) was administered. Patients were divided into two groups. The data of patients whose thumb adduction force was below the median value of all patients were the 'low force' group (9.1 +/- 1.4 N) and the data of all other patients were the 'high force' group (13.7 +/- 1.8 N). RESULTS: In the 'high force' group, maximum neuromuscular blockade of mivacurium was deeper (0.97 +/- 0.05 vs. 0.93 +/- 0.06; P < 0.05) and onset faster (2.9 +/- 1.1 min vs. 4.0 +/- 1.2 min; P < 0.05). Neuromuscular recovery did not differ between the groups. CONCLUSION: The different onset speeds reflect either different sensitivity to neuromuscular blocking agents with respect to patients' muscle power or a problem of the mechanomyographic measuring technique.     

Calibration of histological retina specimens after fixation in Margo’s solution and paraffin embedding to in-vivo dimensions, using photography and optical coherence tomography

Background

The extent of retinal tissue deformation by histological processing needs to be separately measured for every workup protocol. This work presents a simple approach for its quantitative assessment, and shows lateral and axial scaling factors for a common protocol. We calibrated histological measurements by in-vivo photographic and optical coherence tomographic (OCT) measurements, using retinal photocoagulation lesions as calibration markers.

Methods

We evaluated four rabbit eyes that were examined histologically after fixation in Margo’s solution (1 % paraformaldehyde:1.25 % glutaraldehyde), isopropanol dehydration, paraffin embedding and hematoxylin and eosin staining. Distances between 51 pairs of laser lesions were compared in photographs and on histological slides. Retinal thickness measurements were performed at 15 anatomically defined sites in these eyes, and related to anatomically matched OCT thickness measurements of six different rabbit eyes.

Results

We found that the ratio of histological over photographic lesion distances was 1.17 (95 % CI 1.13–1.22), indicating 17 % lateral retinal stretching or expansion by the processing. Thickness measurements in histology were 65.6 % of the in-vivo thickness as measured in OCT, indicating 1/3 axial tissue compression or shrinkage.

Conclusions

We provide an analysis of retinal tissue deformation after fixation in Margo’s solution and paraffin embedding. In spite of protocol optimization for reduced tissue deformation, the workup caused 1/3 axial compression/shrinkage and 17 % lateral elongation, which was unexpected. We show a simple way how to calibrate retina specimens by fundus photography and OCT, two methods that are readily available to most ophthalmologists. Our findings underline the necessity to calibrate specimens prior to morphometry.