[Relevance of big data for molecular diagnostics]. / Bedeutung von Big Data für die molekulare Diagnostik.

Big data analysis raises the expectation that computerized algorithms may extract new knowledge from otherwise unmanageable vast data sets. What are the algorithms behind the big data discussion? In principle, high throughput technologies in molecular research already introduced big data and the development and application of analysis tools into the field of rheumatology some 15 years ago. This includes especially omics technologies, such as genomics, transcriptomics and cytomics. Some basic methods of data analysis are provided along with the technology, however, functional analysis and interpretation requires adaptation of existing or development of new software tools. For these steps, structuring and evaluating according to the biological context is extremely important and not only a mathematical problem. This aspect has to be considered much more for molecular big data than for those analyzed in health economy or epidemiology. Molecular data are structured in a first order determined by the applied technology and present quantitative characteristics that follow the principles of their biological nature. These biological dependencies have to be integrated into software solutions, which may require networks of molecular big data of the same or even different technologies in order to achieve cross-technology confirmation. More and more extensive recording of molecular processes also in individual patients are generating personal big data and require new strategies for management in order to develop data-driven individualized interpretation concepts. With this perspective in mind, translation of information derived from molecular big data will also require new specifications for education and professional competence.

Translational neurocardiology: preclinical models and cardioneural integrative aspects.

Neuronal elements distributed throughout the cardiac nervous system, from the level of the insular cortex to the intrinsic cardiac nervous system, are in constant communication with one another to ensure that cardiac output matches the dynamic process of regional blood flow demand. Neural elements in their various 'levels' become differentially recruited in the transduction of sensory inputs arising from the heart, major vessels, other visceral organs and somatic structures to optimize neuronal coordination of regional cardiac function. This White Paper will review the relevant aspects of the structural and functional organization for autonomic control of the heart in normal conditions, how these systems remodel/adapt during cardiac disease, and finally how such knowledge can be leveraged in the evolving realm of autonomic regulation therapy for cardiac therapeutics.

Postural influence on intracranial and cerebral perfusion pressure in ambulatory neurosurgical patients.

We evaluated postural effects on intracranial pressure (ICP) and cerebral perfusion pressure [CPP: mean arterial pressure (MAP) - ICP] in neurosurgical patients undergoing 24-h ICP monitoring as part of their diagnostic workup. We identified nine patients (5 women, age 44 ± 20 yr; means ± SD), who were "as normal as possible," i.e., without indication for neurosurgical intervention (e.g., focal lesions, global edema, abnormalities in ICP-profile, or cerebrospinal fluid dynamics). ICP (tip-transducer probe; Raumedic) in the brain parenchyma (n = 7) or in the lateral ventricles (n = 2) and cardiovascular variables (Nexfin) were determined from 20° head-down tilt to standing up. Compared with the supine position, ICP increased during 10° and 20° of head-down tilt (from 9.4 ± 3.8 to 14.3 ± 4.7 and 19 ± 4.7 mmHg; P < 0.001). Conversely, 10° and 20° head-up tilt reduced ICP to 4.8 ± 3.6 and 1.3 ± 3.6 mmHg and ICP reached -2.4 ± 4.2 mmHg in the standing position (P < 0.05). Concordant changes in MAP maintained CPP at 77 ± 7 mmHg regardless of body position (P = 0.95). During head-down tilt, the increase in ICP corresponded to a hydrostatic pressure gradient with reference just below the heart, likely reflecting the venous hydrostatic indifference point. When upright, the decrease in ICP was attenuated, corresponding to formation of a separate hydrostatic gradient with reference to the base of the skull, likely reflecting the site of venous collapse. ICP therefore seems to be governed by pressure in the draining veins and collapse of neck veins may protect the brain from being exposed to a large negative pressure when upright. Despite positional changes in ICP, MAP keeps CPP tightly regulated.

[Physiopathology of acute renal failure during sepsis]. / Fisiopatología de la insuficiencia renal aguda durante la sepsis.

Acute renal failure (ARF) is an independent risk factor associated with increased mortality during sepsis. Recent consensus definitions have allowed the standardization of research on the subject. The understanding of the physiopathology of ARF during sepsis is limited by the scarcity of histological studies and the inability to measure renal microcirculatory flows. Historically, ARF during sepsis has been considered to be a consequence of diminished renal blood flow (RBF). Indeed, in early stages of sepsis or in sepsis associated to cardiogenic shock, RBF may decrease. However, recent studies have shown that in resuscitated sepsis, in which cardiac output is characteristically normal or even elevated and there is systemic vasodilatation, RBF is normal or even increased, with no associated histological evidence of significant tubular necrosis. Thus, other factors may participate in the genesis of ARF in sepsis. These include apoptosis, glomerular and medullary microcirculatory disorders, cell changes in response to the pro-inflammatory cascade characteristic of sepsis, oxidative stress, mitochondrial dysfunction and damage induced by mechanical ventilation, among others. Sepsis associated ARF treatment is supportive. In general, renal replacement therapies can be grouped as intermittent or continuous, and as those whose primary objective is the replacement of impaired renal function, versus those whose main objective is to secure hemodynamic stability through the clearing of pro-inflammatory mediators.

Staphtox predictor - A dynamic mathematical model to predict formation of Staphylococcus enterotoxin during heating and fermentation of meat products.

Existing growth models for S. aureus predict growth in relation to temperature, aw/NaCl and pH, and the assessment of probable Staphylococcus enterotoxin (SE) formation is based solely on the number of S. aureus. However, during the production of meat products such as fermented sausages and semi-processed hams, growth of S. aureus is a critical control point in HACCP plans. There is a need to develop a model that evaluates the safety of the product regarding SE formation in relation to the product composition, changes in pH or temperature during the processing and the number of S. aureus in the final product. The objective of the present work is to develop a mathematical model that predicts both the increase in the number of S. aureus and whether SE formation is possible in different meat product processes. A total of 78 experiments were carried out in a meat model system. The experiments covered a range of different temperatures (10-40 °C), pH (4.6-6.0), water phase salt (WPS) (2.2-5.6%) and Sodium nitrite concentrations (0-150 ppm). The meat model system was inoculated with approximately 103 CFU/g of a multi-strain cocktail and incubated at the different temperatures. The cocktail consisted of three strains of S. aureus producing the Staphylococcus enterotoxins A to D (SEA to SED) and a methicillin-resistant strain producing SEG, SEI, SEM, SEN, SEO and SEU. Enumeration of S. aureus was performed several times during the incubation, SE was extracted from samples with >5 log CFU/g, and the SEA-E content was analysed by an ELISA method. Maximum growth rates and lag times calculated from microbiological data, together with temperature, pH, WPS and Sodium nitrite, were used to develop a SE and a growth model. The growth model was developed by training a neural network and the SE model based on logistic regression. The SE and growth models were validated on separate data sets (N = 200 SE model, N = 63 growth model) including both dynamic and static conditions. The SE model predicted all occurrences of toxin formation in the validation data sets. The growth model is a fail-safe model and the prediction errors are comparable to laboratory reproducibility. In conclusion, the models are applicable for predicting the increase in S. aureus and for evaluating if SE formation is likely during processing of meat products. The models are available to producers and other interested parties at www.dmripredict.dk.

Reversible photoswitching enables single-molecule fluorescence fluctuation spectroscopy at high molecular concentration.

We demonstrate that photoswitchable markers enable fluorescence fluctuation spectroscopy at high molecular concentration. Reversible photoswitching allows precise control of the density of fluorescing entities, because the equilibrium between the fluorescent ON- and the dark OFF-state can be shifted through optical irradiation at a specific wavelength. Depending on the irradiation intensity, the concentration of the ON-state markers can be up to 1,000 times lower than the actual concentration of the labeled molecular entity. Photoswitching expands the range of single-molecule detection based experiments such as fluorescence fluctuation spectroscopy to large entity concentrations in the micromolar range.

Rapid baroreceptor resetting in Dahl salt-sensitive rats.

Dahl salt-sensitive rats rapidly become hypertensive when exposed to a high salt diet, but Dahl salt-resistant rats maintain normal blood pressure on a high salt diet. A defect in baroreceptor afferents is thought to play a key role in the low sensitivity of baroreceptor reflexes in Dahl salt-sensitive rats even in the prehypertensive stage during low salt treatment. In the present study, we tested whether differences in rapid resetting ability might contribute to differences in baroreceptor function in Dahl rats. Four groups of rats were tested: salt-sensitive and salt-resistant rats on low salt and high salt diets (0.15% and 8.0% NaCl). We compared the rapidly resetting responses of baroreceptors from each group using an in vitro preparation. Rapid resetting was assessed for each aortic baroreceptor (n = 46) by linear fit of the relation of pressure threshold and conditioning mean arterial pressure. Each group had a wide range of resetting ratios (the slope of the resetting relation). Despite higher initial pressure thresholds in salt-sensitive rats on a high salt diet, resetting ratios among the four groups were similar. Thus, the ability of Dahl salt-sensitive baroreceptors to rapidly reset is preserved, despite high dietary salt and a genetic predisposition to dysfunction. The present findings in Dahl rats reinforce the results of recent studies of rapid resetting during spontaneous and renal hypertension, which suggests that the rapid resetting process is remarkably resistant to factors that compromise baroreceptor function.

Nicotinic activity in the interpeduncular nucleus of the midbrain prolongs recovery from halothane anesthesia.

The influence of nicotinic transmission in the interpeduncular nucleus of the ventral midbrain on recovery from general anesthesia (3% halothane in oxygen) was assessed in rats. Immediately upon withdrawal of the anesthetic, nicotine (2 microliters, 10(-5) to 10(-1) M) was injected into the interpeduncular nucleus. Larger doses of nicotine (10(-2) and 10(-1) M) significantly (P < 0.05) prolonged the recovery of righting reflexes (to 371 +/- 55 and 362 +/- 67 sec, respectively, mean +/- SE), compared with injection of saline (187 +/- 19 sec). Prior intramuscular administration of the nicotinic antagonist, mecamylamine (2 mg/kg) significantly reduced the effect of 10(-2) M nicotine (to 211 +/- 43 sec). Injection of the nicotinic antagonist, hexamethonium (10(-1) M) led to a low mean recovery time (181 +/- 21 sec), not significantly different from control. Prolongation of recovery by 10(-2) M nicotine was not found to be significant when sites more dorsal to the interpeduncular nucleus were injected. An observed tendency for injection of nicotine to slow the post-anesthesia rate of breathing was not statistically significant and not correlated anatomically with the injection site in the midbrain. Increased release of acetylcholine has been shown previously to occur in the interpeduncular nucleus during anesthesia. The present results suggest that nicotinic activation of the interpeduncular nucleus facilitates or sums with the mechanisms in the brain that produce anesthesia under halothane.

Serotonergic, cholinergic and nociceptive inhibition or excitation of raphe magnus neurons in barbiturate-anesthetized rats.

Neurons in the nucleus raphe magnus were recorded extracellularly from barbiturate-anesthetized rats, and were classified by their responses to noxious mechanical stimulation as either pinch-excited, pinch-inhibited or biphasic (inhibited then excited). They were then subjected to iontophoresis of serotonin, some serotonergic agonists and antagonists, acetylcholine, and gamma-amino-n-butyric acid. Serotonin reduced the spontaneous firing of most pinch-inhibited cells (79%). Significantly fewer (P < 0.05) pinch-excited and biphasic cells were inhibited by serotonin (40% and 45%, respectively); in these two cell classes, the observed response was often excitation (30% and 14%), or inhibition for 10-30s followed by excitation for the next 1-2 min (25% and 36%). Acetylcholine showed a similar, statistically significant distribution of effects (P < 0.05), inhibiting all pinch-inhibited neurons (n = 10) but fewer pinch-excited (53%, n = 17) and biphasic neurons (20%, n = 10). Excitation, or excitation then inhibition, was again found frequently among the remaining pinch-excited and biphasic cells. The effect of gamma-amino-n-butyric acid was only inhibitory. In all three nociceptive classes, the serotonin-1A agonist buspirone (n = 15) was inhibitory (87%) and the serotonin-1C/2 antagonist ketanserin (n = 20) was excitatory (35%). The mixed serotonin-1/2 antagonist methysergide (n = 10) was inhibitory (50%) or excitatory (40%). 8-Hydroxy-dipropylaminotetralin (n = 3) was found to increase spontaneous activity (possibly because of partial serotonin-1A agonsim), and +/- propranolol (n = 4) to reduce it (possibly through beta-adrenoceptor antagonism, not serotonin-1A antagonism).(ABSTRACT TRUNCATED AT 250 WORDS)

Morning plasma cortisol levels in infants treated with topical fluorinated glucocorticosteroids.

The plasma cortisol levels of 17 infants of mean age 12.1 months taken at 8 AM were measured before and after application of a topical fluorinated glucocorticosteroid. There was a statistically significant depression of plasma cortisol value in 11 infants at days 3, 7, and 14 after therapy. All of the plasma cortisol levels returned to normal. For the entire group of 17 infants, however, there was no significant depression of plasma cortisol levels. Even in the usual office use of topical glucocorticosteroids, one must be aware of the possiblity of percutaneous absorption.

Cellular mechanisms of baroreceptor integration at the nucleus tractus solitarius.

The autonomic nervous system makes important contributions to the homeostatic regulation of the heart and blood vessels through arterial baroreflexes, and yet our understanding of the central nervous system mechanisms is limited. The sensory synapse of baroreceptors in the nucleus tractus solitarius (NTS) is unique because its participation is obligatory in the baroreflex. Here we describe experiments targeting this synapse to provide greater understanding of the cellular mechanisms at the earliest stages of the baroreflex. Our approach utilizes electrophysiology, pharmacology, and anatomical tracers to identify and evaluate key elements of the sensory information processing in NTS.

Acetylcholine release from the midbrain interpeduncular nucleus during anesthesia.

An exceptional local rise in metabolism during general anesthesia has been noted previously in the ventral midbrain's highly cholinoceptive interpeduncular nucleus (IPN). We report here a functional correlate. Increased interstitial acetylcholine (ACh) was measured in the IPN of rats through chronically implanted microdialysis probes upon anesthesia by inhalation of 3% halothane (mean 1425% of pre-anesthesia baseline at 30 min, n = 5) and by i.p. injection of 100 mg kg-1 ketamine (mean 387%, n = 6). With 50 mg kg-1 i.p. pentobarbital (n = 8), ACh either climbed or fell repeatably in each animal; a positive correlation (p less than 0.05) emerged between the baseline preanesthetized level and the percentage change after 60 min. Mapping of the brainstem under ketamine (n = 2) or pentobarbital (n = 3) anesthesia showed the ACh source to lie in the IPN. We conclude that physiological responses to the chemically and pharmacologically diverse anesthetics halothane and ketamine, and probably also to pentobarbital, converge to enhance the output of the IPN.

Contribution of potassium channels to the discharge properties of rat aortic baroreceptor sensory endings.

The expression of several types of membrane potassium channel at the cell body and central synaptic terminal of the rat aortic arch baroreceptor has been reported by others. It is not known if any of the same channels function at the peripheral sensory terminal of these afferent nerves. Our study examined the effect of three potassium channel blocking agents on the pressure-evoked discharge of such baroreceptors. Thirty-one single unit, regularly discharging baroreceptors were studied using an in vitro aortic arch-aortic nerve preparation. Discharge thresholds and suprathreshold pressure sensitivities were derived from responses of receptors to slowly rising ramps of pressure applied to the aortic arch. Vessel diameter was recorded along with receptor discharge to assess any drug-induced changes in vascular smooth muscle. The blocking agents tested have a range of specificities for classes of potassium channels: tetraethylammonium (TEA), 4-aminopyridine (4-AP) and charybdotoxin. TEA depressed the pressure sensitivity of all baroreceptors tested (n = 3) in a dose-dependent manner. Baroreceptor responses to 4-AP were complex (n = 22) and varied widely across individuals. Three were unaffected by 5 mM 4-AP. Most baroreceptors were generally depressed by 4-AP. Some of the 4-AP effects appeared to be related to actions at vascular smooth muscle. None of the baroreceptors tested (n = 6) was affected by charybdotoxin. The results of selective potassium channel blockade are generally consistent with what would be expected from a sustained depolarization of baroreceptor endings such as has been reported with raising extracellular potassium and probably includes effects of inactivation of other voltage-dependent channels.(ABSTRACT TRUNCATED AT 250 WORDS)

Localization and retention in vitro of fluorescently labeled aortic baroreceptor terminals on neurons from the nucleus tractus solitarius.

The anterograde fluorescent tracer DiA was used to visualize baroreceptor fibers and synaptic terminals both in living and fixed tissue. Baroreceptor fibers labeled with DiA terminated as a dense synaptic field in the medial nucleus tractus solitarius (NTS), making synaptic contact on the soma, as well as processes of neurons that they innervated. A similar distribution and morphology was observed in baroreceptor fibers and terminals labeled with horseradish peroxidase. DiA also identified baroreceptor terminals and the neurons receiving these synaptic contacts in vitro. NTS neurons were dissociated from their surrounding tissue and identified by attached baroreceptor terminals that retained the fluorescent dye. These results will enable us to study the electrophysiological properties of dispersed neurons that receive identified baroreceptor synaptic terminals.

Use of methylene blue in patients with refractory septic shock: impact on hemodynamics and gas exchange.

PURPOSE: The purpose of this study was to assess the acute effects of methylene blue, an inhibitor of nitric oxide synthesis, on hemodynamics and gas exchange in patients with refractory septic shock in a prospective clinical trial at medical and surgical intensive care units in a tertiary university hospital. PATIENTS AND METHODS: Prospective, sequential study of 10 consecutive patients admitted with severe septic shock of diverse causes and unable to achieve an adequate arterial pressure despite the use of at least two vasoactive drugs. Six of them also developed acute lung injury. All received 1 mg/kg intravenous bolus of methylene blue. Hemodynamic and respiratory parameters were measured at baseline and at 30, 60, 120, and 180 minutes after the bolus injection. RESULTS: Systolic, diastolic, mean arterial blood pressure, and systemic vascular resistance increased significantly in all patients, whereas no significant changes were observed in cardiac output, oxygen consumption, or oxygen extraction ratio. Gas exchange remained unaffected in patients with acute lung injury. CONCLUSIONS: Methylene blue had an acute vasopressor effect in patients with refractory septic shock, and it was not deleterious on respiratory function.

Differentiation of autonomic reflex control begins with cellular mechanisms at the first synapse within the nucleus tractus solitarius.

Visceral afferents send information via cranial nerves to the nucleus tractus solitarius (NTS). The NTS is the initial step of information processing that culminates in homeostatic reflex responses. Recent evidence suggests that strong afferent synaptic responses in the NTS are most often modulated by depression and this forms a basic principle of central integration of these autonomic pathways. The visceral afferent synapse is uncommonly powerful at the NTS with large unitary response amplitudes and depression rather than facilitation at moderate to high frequencies of activation. Substantial signal depression occurs through multiple mechanisms at this very first brainstem synapse onto second order NTS neurons. This review highlights new approaches to the study of these basic processes featuring patch clamp recordings in NTS brain slices and optical techniques with fluorescent tracers. The vanilloid receptor agonist, capsaicin, distinguishes two classes of second order neurons (capsaicin sensitive or capsaicin resistant) that appear to reflect unmyelinated and myelinated afferent pathways. The differences in cellular properties of these two classes of NTS neurons indicate clear functional differentiation at both the pre- and postsynaptic portions of these first synapses. By virtue of their position at the earliest stage of these pathways, such mechanistic differences probably impart important differentiation in the performance over the entire reflex pathways.

Liquid chromatographic determination of progestogens in animal fat.

A liquid chromatographic method was developed for determination of 3 progestogens-melengestrol acetate, megestrol acetate, and chlormadinone acetate-found in edible tissue at concentrations between 10 and 1000 ppb. These progestogens are commonly used as feed additives to control herd estrus and to improve feed efficiency. Rendered fat was extracted with acetonitrile, washed with hexane, and dried. The remaining lipids were saponified with sodium hydroxide and precipitated with magnesium chloride. The progestogens were extracted from the basic solution with hexane, dried, and cleaned up on a cyanopropyl solid-phase extraction column in the normal-phase mode. The eluate was dried and reconstituted with acetonitrile-water (7 + 3, v/v). Chromatography was performed on a 5 microns high-carbon load C18 column with acetonitrile-water (7 + 3, v/v) at 1 mL/min and UV detection at 291 nm. Recoveries from fortified samples ranged from 84 to 116%. The limit of quantitation was 10 ppb for both beef and pork. The detection limit was 3 ppb.

Academic progress of children who attended a preschool for abused children: a follow-up of the keepsafe project.

OBJECTIVE: This study was undertaken to assess academic progress of children 1 to 5 years after graduating from the C. Henry Kempe Center's Therapeutic Preschool Day Treatment Program. METHOD: Information was gathered through chart review; telephone surveys of care providers, relatives, and social service workers; as well questionnaires on all children who attended the therapeutic preschool day treatment program between 1984 and 1989, including the 24 children reported on by Oates, Gray, Schweitzer, Kempe, and Harmon, 1995. RESULTS: Classroom placement was determined for 27 of the 44 graduates (61.4%), 14 of whom (51.9%) were in a regular classroom, 10 (37.0%) in special education, two (7.4%) in residential treatment, and one (3.7%) was receiving home schooling. Twenty-two of the 27 children (81.5%) improved or remained in the same grade and type of classroom as they had been staffed into at the time of their graduation from the preschool day treatment program. Factors thought to affect stability of classroom placement were studied, of which frequency of family moves was the only significant variable. Its significance was in the direction opposite to that expected. The most effective method of locating families was to contact the Department of Social Services who provided information used to find 60% of them. CONCLUSIONS: Intervention in a therapeutic preschool day treatment program was found to be beneficial, as it enabled most of the children to progress appropriately in public school.

[Cost analysis of dialysis treatment at the Odense University Hospital and the Sønderborg Hospital]. / Omkostningsanalyse af dialysebehandlingen på Odense Universitetshospital og Sønderborg Sygehus.

The major purpose of this paper is to investigate the treatment costs of dialysis treatment by modality. In this study Odense University Hospital (OUH) and Sønderborg Hospital were chosen as cases. The costs of haemodialysis (HD) treatment are estimated to DKK 341-392,000 per patient during the first year, and DKK 328-379,000 per year the following years. The costs of continuous ambulatory peritoneal dialysis (CAPD) treatment are estimated to DKK 262-291,000 per patient during the first year, and DKK 251-277,000 per year the following years. The costs of CCPD (peritoneal dialysis with the aid of a machine), treatment are estimated to DKK 312-325,000 per patient during the first year, and DKK 296-308,000 per year the following years. The treatment costs of HD are lower than expected, while the treatment costs of PD are higher than expected. As a result of this the differences in treatment costs (HD versus PD) are much lower than expected, DKK 130,000 at the most.

Intracranial Pressure Monitoring: Invasive versus Non-Invasive Methods-A Review.

Monitoring of intracranial pressure (ICP) has been used for decades in the fields of neurosurgery and neurology. There are multiple techniques: invasive as well as noninvasive. This paper aims to provide an overview of the advantages and disadvantages of the most common and well-known methods as well as assess whether noninvasive techniques (transcranial Doppler, tympanic membrane displacement, optic nerve sheath diameter, CT scan/MRI and fundoscopy) can be used as reliable alternatives to the invasive techniques (ventriculostomy and microtransducers). Ventriculostomy is considered the gold standard in terms of accurate measurement of pressure, although microtransducers generally are just as accurate. Both invasive techniques are associated with a minor risk of complications such as hemorrhage and infection. Furthermore, zero drift is a problem with selected microtransducers. The non-invasive techniques are without the invasive methods' risk of complication, but fail to measure ICP accurately enough to be used as routine alternatives to invasive measurement. We conclude that invasive measurement is currently the only option for accurate measurement of ICP.