P29: a novel tyrosine-phosphorylated membrane protein present in small clear vesicles of neurons and endocrine cells.

A novel membrane protein from rat brain synaptic vesicles with an apparent 29,000 Mr (p29) was characterized. Using monospecific polyclonal antibodies, the distribution of p29 was studied in a variety of tissues by light and electron microscopy and immunoblot analysis. Within the nervous system, p29 was present in virtually all nerve terminals. It was selectively associated with small synaptic vesicles and a perinuclear region corresponding to the area of the Golgi complex. P29 was not detected in any other subcellular organelles including large dense-core vesicles. The distribution of p29 in various subcellular fractions from rat brain was very similar to that of synaptophysin and synaptobrevin. The highest enrichment occurred in purified small synaptic vesicles. Outside the nervous system, p29 was found only in endocrine cell types specialized for peptide hormone secretion. In these cells, p29 had a distribution very similar to that of synaptophysin. It was associated with microvesicles of heterogeneous size and shape that are primarily concentrated in the centrosomal-Golgi complex area. Secretory granules were mostly unlabeled, but their membrane occasionally contained small labeled evaginations. Immunoisolation of subcellular organelles from undifferentiated PC12 cells with antisynaptophysin antibodies led to a concomitant enrichment of p29, synaptobrevin, and synaptophysin, further supporting a colocalization of all three proteins. P29 has an isoelectric point of approximately 5.0 and is not N-glycosylated. It is an integral membrane protein and all antibody binding sites are exposed on the cytoplasmic side of the vesicles. Two monoclonal antibodies raised against p29 cross reacted with synaptophysin, indicating the presence of related epitopes. P29, like synaptophysin, was phosphorylated on tyrosine residues by endogenous tyrosine kinase activity in intact vesicles.

The effect of orthostasis on recurrence quantification analysis of heart rate and blood pressure dynamics.

The purpose of this paper is to investigate the effect of orthostatic challenge on recurrence plot based complexity measures of heart rate and blood pressure variability (HRV and BPV). HRV and BPV complexities were assessed in 28 healthy subjects over 15 min in the supine and standing positions. The complexity of HRV and BPV was assessed based on recurrence quantification analysis. HRV complexity was reduced along with the HRV magnitude after changing from the supine to the standing position. In contrast, the BPV magnitude increased and BPV complexity decreased upon standing. Recurrence quantification analysis (RQA) of HRV and BPV is sensitive to orthostatic challenge and might therefore be suited to assess changes in autonomic neural outflow to the cardiovascular system.

A synaptic vesicle membrane protein is conserved from mammals to Drosophila.

The structure of synaptobrevin, an intrinsic membrane protein of small synaptic vesicles from mammalian brain, was studied by purification and molecular cloning. Its message in bovine brain encodes a 116 amino acid protein whose sequence reveals it to be the mammalian homolog of Torpedo VAMP-1. Antibody probing demonstrates that the protein is also present in Drosophila, and its Drosophila homolog was cloned. Alignment of the sequences of synaptobrevin/VAMP-1 from the three species shows it to contain four domains, including a highly conserved central region of 63 amino acids that contains 75% invariant residues. The finding that a membrane protein from vertebrate synaptic vesicles is conserved in Drosophila points toward a central role of this protein in neurotransmission and should allow a genetic approach to neurotransmitter release.

Synaptic vesicles immunoisolated from rat cerebral cortex contain high levels of glutamate.

L-Glutamate is regarded as the major excitatory neurotransmitter in the mammalian CNS. However, whether the released transmitter originates from a cytosolic pool or is discharged from synaptic vesicles by exocytosis (vesicle hypothesis) remains controversial. A problem with the general acceptance of the vesicle hypothesis is that the enrichment of glutamate in synaptic vesicles has not been convincingly demonstrated. In the present study, we have analyzed the glutamate content of synaptic vesicles isolated from rat cerebral cortex by a novel immunobead procedure. A large amount of glutamate was present in these vesicles when a proton electrochemical gradient was maintained across the vesicle membrane during isolation. Compared with the starting fraction, glutamate was enriched more than 10-fold relative to other amino acids. Addition of N-ethylmaleimide prevented glutamate loss during isolation. Isotope exchange experiments revealed that exchange or re-uptake of glutamate after homogenization is negligible. We conclude that rat brain synaptic vesicles contain high levels of glutamate in situ.

Reduced short-term complexity of heart rate and blood pressure dynamics in patients with diabetes mellitus type 1: multiscale entropy analysis.

Multiscale entropy (MSE) analysis provides information about complexity on various time scales. The aim of this study was to test whether MSE is able to detect autonomic dysregulation in young patients with diabetes mellitus (DM). We analyzed heart rate (HR) oscillations, systolic (SBP) and diastolic blood pressure (DBP) signals in 14 patients with DM type 1 and 14 age- and sex-matched healthy controls. SampEn values (scales 1-10) and linear measures were computed. HR: among the linear measures of heart rate variability significant differences between groups were only found for RMSSD (p = 0.043). MSE was significantly reduced on scales 2 and 3 in DM (p = 0.023 and 0.010, respectively). SBP and DBP: no significant differences were detected with linear measures. In contrast, MSE analysis revealed significantly lower SampEn values in DM on scale 3 (p = 0.039 for SBP; p = 0.015 for DBP). No significant correlations were found between MSE and linear measures. In conclusion, MSE analysis of HR, SBP and DBP oscillations is able to detect subtle abnormalities in cardiovascular control in young patients with DM and is independent of standard linear measures.

Pulse wave amplitude and heart period variability in children with upper airway obstruction.

OBJECTIVE: The aim of this study was to assess cardiovascular autonomic modulation in children with upper airway obstruction (UAO), to compare this modulation to that of non-snoring children and to investigate the effect of adenotonsillectomy (AT). METHODS: ECG and finger photoplethysmographic signals obtained from overnight polysomnographic (PSG) recordings of 31 children with mild-to-moderate UAO and 34 non-snoring children were analysed. The extent of autonomic modulation was assessed by symbolic analysis of heart period (HP), pulse wave amplitude (PWA), and their joint dynamics during non-rapid eye movement (NREM) sleep and rapid eye movement (REM) sleep. RESULTS: Children with UAO showed more frequent patterns of monotonically increasing and decreasing HP in NREM sleep and monotonically increasing and decreasing joint PWA-HP patterns in REM and NREM sleep at baseline compared to controls, even when considering only periods of sleep free of discrete respiratory events. Following AT, HP, and joint PWA-HP dynamics significantly altered towards levels observed in the control group. CONCLUSIONS: In children with mild-to-moderate UAO, cardiovascular autonomic modulation is more prevalent, even during quiet, event-free sleep. AT appears to reverse this pattern.

Hidden Markov models based on symbolic dynamics for statistical modeling of cardiovascular control in hypertensive pregnancy disorders.

Discrete hidden Markov models (HMMs) were applied to classify pregnancy disorders. The observation sequence was generated by transforming RR and systolic blood pressure time series using symbolic dynamics. Time series were recorded from 15 women with pregnancy-induced hypertension, 34 with preeclampsia and 41 controls beyond 30th gestational week. HMMs with five to ten hidden states were found to be sufficient to characterize different blood pressure variability, whereas significant classification in RR-based HMMs was found using fifteen hidden states. Pregnancy disorders preeclampsia and pregnancy induced hypertension revealed different patho-physiological autonomous regulation supposing different etiology of both disorders.

T-wave morphology can distinguish healthy controls from LQTS patients.

Long QT syndrome (LQTS) is an inherited disorder associated with prolongation of the QT/QTc interval on the surface electrocardiogram (ECG) and a markedly increased risk of sudden cardiac death due to cardiac arrhythmias. Up to 25% of genotype-positive LQTS patients have QT/QTc intervals in the normal range. These patients are, however, still at increased risk of life-threatening events compared to their genotype-negative siblings. Previous studies have shown that analysis of T-wave morphology may enhance discrimination between control and LQTS patients. In this study we tested the hypothesis that automated analysis of T-wave morphology from Holter ECG recordings could distinguish between control and LQTS patients with QTc values in the range 400-450 ms. Holter ECGs were obtained from the Telemetric and Holter ECG Warehouse (THEW) database. Frequency binned averaged ECG waveforms were obtained and extracted T-waves were fitted with a combination of 3 sigmoid functions (upslope, downslope and switch) or two 9th order polynomial functions (upslope and downslope). Neural network classifiers, based on parameters obtained from the sigmoid or polynomial fits to the 1 Hz and 1.3 Hz ECG waveforms, were able to achieve up to 92% discrimination between control and LQTS patients and 88% discrimination between LQTS1 and LQTS2 patients. When we analysed a subgroup of subjects with normal QT intervals (400-450 ms, 67 controls and 61 LQTS), T-wave morphology based parameters enabled 90% discrimination between control and LQTS patients, compared to only 71% when the groups were classified based on QTc alone. In summary, our Holter ECG analysis algorithms demonstrate the feasibility of using automated analysis of T-wave morphology to distinguish LQTS patients, even those with normal QTc, from healthy controls.

QT variability improves risk stratification in patients with dilated cardiomyopathy.

Recently it could be demonstrated that systolic and diastolic blood pressure variability (BPV) as well as segmented Poincare plot analysis (SPPA) contribute to risk stratification in patients suffering from dilated cardiomyopathy (DCM). The aim of this study was to improve the risk stratification applying a multivariate technique including QT variability (QTV). We enrolled and significantly separated 56 low risk and 13 high risk DCM patients by nearly all applied BPV and QTV methods, but not with traditional heart rate variability analysis. The optimum set of two indices calculating the multivariate discriminate analysis (DA) included one BPV index calculated by symbolic dynamics method (DBP(Shannon)) and one index calculated from QTV (QTV(log)) achieving an area under the receiver operating characteristics curve (AUC) of 92%, sensitivity of 92.3% and specificity of 89.3%. Performing only electrocardiogram analysis, the optimum multivariate approach including indices from segmented Poincaré plot analysis and QTV still achieved a remarkable AUC of 88.3%. Increasing the number of indices for multivariate DA up to three, we achieved an AUC of 95.7%, sensitivity of 100% and specificity of 85.7% including one clinical, one BPV and one QTV index. Summarizing, we identified DCM patients with an increased risk of sudden cardiac death applying QTV analysis in a multivariate approach.

Baroreflex sensitivity, heart rate, and blood pressure variability in hypertensive pregnancy disorders.

Hypertensive pregnancy disorders are a leading cause of perinatal and maternal morbidity and mortality. Heart rate variability (HRV), blood pressure variability (BPV), and baroreflex sensitivity (BRS) are relevant predictors of cardiovascular risk in humans. The aim of the study was to evaluate whether HRV, BPV, and BRS differ between distinct hypertensive pregnancy disorders. Continuous heart rate and blood pressure recordings were performed in 80 healthy pregnant women as controls (CON), 19 with chronic hypertension (CH), 18 with pregnancy-induced hypertension (PIH), and 44 with pre-eclampsia (PE). The data were assessed by time and frequency domain analysis, nonlinear dynamics, and BRS. BPV is markedly altered in all three groups with hypertensive disorders compared to healthy pregnancies, whereby changes were most pronounced in PE patients. Interestingly, this increase in PE patients did not lead to elevated spontaneous baroreflex events, while BPV changes in both the other hypertensive groups were paralleled by alterations in baroreflex parameters. The HRV is unaltered in CH and PE but significantly impaired in PIH. We conclude that parameters of the HRV, BPV, and BRS differ between various hypertensive pregnancy disorders. Thus, distinct clinical manifestations of hypertension in pregnancy have different pathophysiological, regulatory, and compensatory mechanisms.

Forecasting of life threatening arrhythmias using the compression entropy of heart rate.

OBJECTIVES: Ventricular tachycardia (VT) provoking sudden cardiac death (SCD) are a major cause of mortality in the developed countries. The most efficient therapy for SCD prevention are implantable cardioverter defibrillators (ICD). In this study heart rate variability (HRV) measures were analyzed for short-term forecasting of VT in order to improve VT sensing and to enable a patient warning of forth-coming shocks. METHODS: The last 1000 normal beat-to-beat intervals before 50 VT episodes stored by the ICD were analyzed and compared to individually acquired control time series (CON). HRV analysis was performed with standard parameters of time and frequency domain as suggested by the HRV Task Force and furthermore with a newly developed and optimized nonlinear parameter that assesses the compression entropy of heart rate (Hc). RESULTS: Except of meanNN (p = 0.02) we found no significant differences in standard HRV parameters. In contrast, Hc revealed highly significant (p = 0.007) alterations in VT compared with CON suggesting a decreased complexity before the onset of VT. CONCLUSION: Compression entropy might be a suitable parameter for short-term forecasting of life-threatening tachycardia in ICD.

Comparative study of neutrophil activities in adults and full-term neonates in relation to the method of delivery.

The aim of this study was to compare the markers of neutrophil activity in healthy adults with those in healthy neonates in relation to the method of delivery. The following parameters were studied: absolute neutrophil count, neutrophil adherence to nylon fiber, NBT reduction, and neutrophil phagocytosis of latex particles. The study has shown that the method of delivery significantly affects the markers of neutrophil activity measured in umbilical cord blood. The levels of these markers were higher in neonates born by elective cesarean section than in adults, neonates born vaginally or those born by emergency cesarean section. This comparative study of defensive functions of neutrophils from adults and neonates indicates that the conflicting results of other studies may be caused not only by differences in investigative methodology but also by the fact that the eligibility criteria used in these studies did not include the method of delivery, which can affect the markers of neutrophil activity via triggering a stress response.

Joint symbolic dynamic analysis of beat-to-beat interactions of heart rate and systolic blood pressure in normal pregnancy.

Pregnancy induces important changes in the autonomic control. Measures of heart rate (HR) variability and systolic blood pressure (SP) variability are sensitive to those changes. The interactions between HR and SP are complex and strongly non-linear. Therefore they cannot be completely described by linear analysis techniques. A study of joint symbolic dynamics is presented as a new short-term non-linear analysis method to investigate the interactions between HR and SP. Continuous, non-invasive 30 min blood pressure recordings (Portapres) of 25 pregnant and 14 non-pregnant women were analysed. Time series of beat-to-beat HR and SP were extracted. Using the concept of joint symbolic dynamics, HR and SP changes were transformed into a bivariate symbol vector. Subsequently, this symbol vector was transformed into a word series (words consisting of three successive symbols), and the probability of occurrence of each word type was calculated and compared between both groups. Significant differences were found in five word types between pregnant and non-pregnant women: w0,4(0.021+/-0.011 against 0.008+/-0.006; p = 0.022), w4,6(0.020+/-0.010 against 0.007+/-0.003; p = 0.001), w3,2(0.004+/-0.003 against 0.007+/-0.003; p = 0.038), W6,5(0.009+/-0.007 against 0.023+/-0.008; p< 0.001) and w3,6(0.011+/-0.007 against 0.023+/-0.008; p = 0.001). Joint symbolic dynamics provides an efficient non-linear representation of HR and SP interactions that offers simple physiological interpretations.

[Heart rate and blood pressure interaction in normotensive and chronic hypertensive pregnancy]. / Herzfrequenz- und Blutdruckinteraktion bei normotensiver und chronisch hypertensiver Schwangerschaft.

Pregnancy has impact on autonomic control. Since hypertensive pregnancy disorders are a major cause of maternal mortality we investigated the baroreflex sensitivity (BRS) in chronic hypertensive pregnant women (CH), normotensive pregnant (PRE) and non-pregnant (NPRE) women. In addition to the traditional 'sequence method' we used joint symbolic dynamics (JSD). BRS was significantly reduced in all pregnancies compared with NPRE (p < 0.00001) but there was no significant difference between CH and PRE. Contrary, the JSD measures revealed significant differences (p < 0.00001) in the heart rate and blood pressure interactions between PRE and CH. In conclusion, JSD measures uncovered a different gestation related adaptation of autonomic regulation in women with chronic.

[Modified interaction of blood pressure and heart rate in idiopathic dilated cardiomyopathy]. / Veränderte Interaktion von Blutdruck und Herzfrequenz bei idiopathischer dilatativer Kardiomyopathie.

BACKGROUND: Neurovegetative and haemodynamic changes impact on the regulation pattern of blood pressure and heart rate in patients with heart failure. We studied these patterns and their interactions in patients with idiopathic dilated cardiomyopathy (IDC) and in healthy subjects (REF). METHODS: We continually measured the heart rate and blood pressure (Portapres device) in twenty-five supine IDC patients (age: 51 +/- 13 y; left ventricular end-diastolic diameter 67 +/- 11 mm; ejection fraction 30 +/- 11%) and in twenty-seven REF (age: 50 +/- 11 y) Recording time was 30 minutes. The heart rate (HR) of each beat and the systolic blood pressure (SYS) of the subsequent beat were measured. Code numbers (symbols) were assigned to the beat-to-beat changes in HR and SYS (increase: 1; decrease: 0). The frequencies of the symbols sequences of three successive beats were counted. In this way we obtained a matrix consisting of eight (two to the power of three) HR and SYS combinations: 000, 100, 010, 001, 111, 110, 011 and 101. We then counted the frequencies of the different combinations of the symbol sequences in HR and SYS (2(3) x 2(3) = 64 combinations). The relative frequencies of symbol patterns appearing in HR, SYS and in the combined analysis of HR and SYS, were compared for IDC and REF using the T-test for independent samples. RESULTS: Significant differences were seen between IDC and REF. The HR patterns 101 and 010 were more frequent in IDC than in REF patients (11.1 +/- 4.7 vs. 7.7 +/- 2.9%, p = 0.003, and 16.1 +/- 6.3 vs. 11.7 +/- 4.9%, p = 0.008). This finding was even more marked in the analysis of the SYS patterns 101 and 010 (11.0 +/- 7.4 vs. 8.2 +/- 2.9%, p < 0.001, and 11.6 +/- 7.4 vs. 5.4 +/- 2.7%, p < 0.001). Non-alternating patterns were more frequent in REF (e.g. 000HR & 111SYS: 4.6 +/- 3.3 vs. 2.9 +/- 2.4%, p = 0.03). CONCLUSIONS: We demonstrated significant interaction of the regulation patterns of blood pressure and heart rate, as also their interactions in IDC. Opposed changes in HR and SYS mediated by the baroreflex, became superimposed by alternans phenomena in IDC. The pattern analysis of changes in HR and SYS detects these disturbances of neurovegetative short-term control.

[Afterload and blood pressure amplitude in dilated cardiomyopathy]. / Nachlast und Blutdruckamplitude bei dilatativer Kardiomyopathie.

The beat-to-beat variability of the diastolic blood pressure induces small variations in the afterload of the left ventricle. These variations influence myocardial contractility, and thus blood pressure amplitude. We assessed the interdependence of blood pressure and changes in the afterload. We continuously recorded blood pressure (duration 200 s, at rest) in 20 patients with dilated cardiomyopathy (ejection fraction 32 +/- 13%, left ventricular diameter 67 +/- 8 mm) and in 20 healthy volunteers. Interbeat intervals, diastolic pressures, systolic pressure amplitudes and mean slopes of systolic pressure amplitudes were measured. Correlation coefficients (r) were calculated to assess the interdependence of blood pressure amplitudes/mean systolic slopes and the preceding diastolic pressures/interbeat intervals, respectively. In healthy volunteers we found a strong interdependence between blood pressure amplitude and the preceding diastolic pressures (r = 0.62 +/- 0.21 and 0.47 +/- 0.22). Higher diastolic pressures were followed by higher blood pressure amplitudes, and by steeper slopes of the systolic peaks. In patients with dilated cardiomyopathy, such interdependence was significantly lower (r = 0.33 +/- 22 and r = 0.28 +/- 0.35), and in patients with severely reduced left ventricular function (ejection fraction < 32%) was only marginal (r = 0.23 +/- 0.27 and 0.21 +/- 0.44, respectively). The forces of the isovolumetric contraction necessary to initiate the ejection phase of the left ventricle depend on the afterload, i.e. on the diastolic pressure. The responses of amplitude and slope of the systolic blood pressure to small changes in the afterload make it possible to assess left ventricular contractility. The latter is impaired in dilated cardiomyopathy.

[Level of cortisol in blood serum of maternal and umbilical cord during various methods of delivery]. / Stezenie kortyzolu w surowicy krwi matki i krwi pepowinowej w róznych sposobach zakonczenia ciazy.

The effect of stress during labor on the plasma concentration of cortisol was studied in 82 women. The total cortisol levels in the maternal serum and cord blood were measured by radioimmunoassay during spontaneous labor (PSN), oxytocin induced labor (PI), elective cesarean section (CCE), and emergency cesarean section (CCC). The result indicate that cortisol level was higher after PI (710 ng/ml) and CCC (755.4 ng/ml) and lower in CCE (278.9 ng/ml) in the maternal vein. In cord blood after CCC was higher (530.8 ng/ml), and lower after CCE (157.8 ng/ml).

[Endothelin and nitric oxide in newborn kidney]. / Fizjologiczna rola endoteliny i tlenku azotu w nerce noworodka.

The review addresses two of vasoactive substances: endothelin and nitric oxide and describes how each of them effects the functions of endothelial cells of kidneys in newborn.

[Evaluation of glucose and cholesterol levels in the mother and newborn infant in the perinatal period]. / Ocena poziomu glukozy i cholesterolu u matki i noworodka w okresie okoloporodowym.

In 30 live born newborns the connections were assessed between glucose and cholesterol levels in umbilical cord blood after birth, and also in maternal blood before and two days after birth. Statistically significant differences of glucose and cholesterol levels were found between mothers and newborns in the perinatal period. Attention is called to greater independence of cholesterol (total, free and esterified) levels in newborn and mother in perinatal period.

[Biological developmental parameters in newborns in Silesia born between the 30th and 42nd week of pregnancy]. / Wskazniki biologiczne rozwoju noworodków slaskich urodzonych miedzy 30 a 42 tygodniem ciazy.

On the basis of anthropometric data from 2985 newborns in Silesia, percentile grids were constructed of birth weight, height, head circumference, and chest circumference for boys, girls, and both sexes jointly. It was found that these parameters, especially in premature babies, were lower in comparison to those accepted for the whole country.