Cholinergic mechanisms in spinal locomotion-potential target for rehabilitation approaches.

Previous experiments implicate cholinergic brainstem and spinal systems in the control of locomotion. Our results demonstrate that the endogenous cholinergic propriospinal system, acting via M2 and M3 muscarinic receptors, is capable of consistently producing well-coordinated locomotor activity in the in vitro neonatal preparation, placing it in a position to contribute to normal locomotion and to provide a basis for recovery of locomotor capability in the absence of descending pathways. Tests of these suggestions, however, reveal that the spinal cholinergic system plays little if any role in the induction of locomotion, because MLR-evoked locomotion in decerebrate cats is not prevented by cholinergic antagonists. Furthermore, it is not required for the development of stepping movements after spinal cord injury, because cholinergic agonists do not facilitate the appearance of locomotion after spinal cord injury, unlike the dramatic locomotion-promoting effects of clonidine, a noradrenergic α-2 agonist. Furthermore, cholinergic antagonists actually improve locomotor activity after spinal cord injury, suggesting that plastic changes in the spinal cholinergic system interfere with locomotion rather than facilitating it. Changes that have been observed in the cholinergic innervation of motoneurons after spinal cord injury do not decrease motoneuron excitability, as expected. Instead, the development of a "hyper-cholinergic" state after spinal cord injury appears to enhance motoneuron output and suppress locomotion. A cholinergic suppression of afferent input from the limb after spinal cord injury is also evident from our data, and this may contribute to the ability of cholinergic antagonists to improve locomotion. Not only is a role for the spinal cholinergic system in suppressing locomotion after SCI suggested by our results, but an obligatory contribution of a brainstem cholinergic relay to reticulospinal locomotor command systems is not confirmed by our experiments.

Influence of substrate nitridation temperature on epitaxial alignment of GaN nanowires to Si(111) substrate.

An arrangement of self-assembled GaN nanowires (NWs) grown by plasma-assisted molecular beam epitaxy on a Si(111) substrate is studied as a function of the temperature at which the substrate is nitridized before GaN growth. We show that the NWs grow with the c-axis perpendicular to the substrate surface independently of nitridation temperature with only a slight improvement in tilt coherency for high nitridation temperatures. A much larger influence of the substrate nitridation process on the in-plane arrangement of NWs is found. For high (850 °C) and medium (450 °C) nitridation temperatures angular twist distributions are relatively narrow and NWs are epitaxially aligned to the substrate in the same way as commonly observed in GaN on Si(111) planar layers with an AlN buffer. However, if the substrate is nitridized at low temperature (~150 °C) the epitaxial relationship with the substrate is lost and an almost random in-plane orientation of GaN NWs is observed. These results are correlated with a microstructure of silicon nitride film created on the substrate as the result of the nitridation procedure.

Commissural interneurons with input from group I and II muscle afferents in feline lumbar segments: neurotransmitters, projections and target cells.

The aim of this study was to analyse neurotransmitter content, projection areas and target cells of commissural interneurons with input from group I and/or II muscle afferents in lumbar segments in the cat. Axonal projections of 15 intracellularly labelled commissural interneurons were reconstructed. Ten interneurons (nine located in laminae VI-VII, one in lamina VIII) were glutamatergic; only one interneuron (located in lamina VIII) was glycinergic. Contralateral terminal projections were found both in motor nuclei and within laminae VI-VIII. In order to identify target cells of commissural interneurons, effects of stimulation of contralateral group I and II muscle afferents were investigated on interneurons within these laminae. Three tests were used: intracellular records from individual interneurons, modulation of probability of activation of extracellularly recorded interneurons and modulation of their actions on motoneurons using disynaptic PSPs evoked in motoneurons as a measure. All these tests revealed much more frequent and/or stronger excitatory actions of contralateral afferents. The results indicate that commissural interneurons with input from contralateral group I and II afferents target premotor interneurons in disynaptic pathways from ipsilateral group I and II afferents and that excitatory disynaptic actions of contralateral afferents on these interneurons are mediated primarily by intermediate zone commissural interneurons. A second group of commissural interneurons activated by reticulospinal neurons, previously described, frequently had similar, but occasionally opposing, actions to the cells described here, thus indicating that these two subpopulations may act on the same premotor interneurons and either mutually enhance or counteract each other's actions.

Premotor interneurones contributing to actions of feline pyramidal tract neurones on ipsilateral hindlimb motoneurones.

The aim of the study was to analyse the potential contribution of excitatory and inhibitory premotor interneurones in reflex pathways from muscle afferents to actions of pyramidal tract (PT) neurones on ipsilateral hindlimb motoneurones. Disynaptic EPSPs and IPSPs evoked in motoneurones in deeply anaesthetized cats by group Ia, Ib and II muscle afferents were found to be facilitated by stimulation of the ipsilateral, as well as of contralateral, PT. The ipsilateral actions were evoked by either uncrossed or double-crossed pathways. The results show that interneurones mediating reflex actions of muscle afferents may be activated strongly enough by PT stimulation to contribute to movements initiated by ipsilateral PT neurones and that PT actions relayed by them might be enhanced by muscle stretches and/or contractions. However, in some motoneurones disynaptic IPSPs and EPSPs evoked from group Ib or II afferents were depressed by PT stimulation. In order to analyse the basis of this depression, the transmitter content in terminals of 11 intracellularly labelled interneurones excited by PT stimulation was defined immunohistochemically and their axonal projections were reconstructed. The interneurones included 9 glycinergic and 2 glutamatergic neurones. All but one of these neurones were mono- or disynaptically excited by group I and/or II afferents. Several projected to motor nuclei and formed contacts with motoneurones. However, all had terminal projections to areas outside the motor nuclei. Therefore both inhibitory and excitatory interneurones could modulate responses of other premotor interneurones in parallel with direct actions on motoneurones.

Neuronal relays in double crossed pathways between feline motor cortex and ipsilateral hindlimb motoneurones.

Coupling between pyramidal tract (PT) neurones and ipsilateral hindlimb motoneurones was investigated by recording from commissural interneurones interposed between them. Near maximal stimulation of either the left or right PT induced short latency EPSPs in more than 80% of 20 commissural interneurones that were monosynaptically excited by reticulospinal tract fibres in the medial longitudinal fascicle (MLF). The EPSPs were evoked at latencies that were only 1-2 ms longer than those of EPSPs evoked from the MLF, compatible with a disynaptic coupling between PT fibres and these commissural interneurones. EPSPs evoked by PT stimulation were frequently associated with IPSPs which either followed or preceded the EPSPs. The latencies of the IPSPs (on average about 1 ms longer than latencies of the earliest EPSPs) indicated that they were mediated via single additional inhibitory interneurones. Records from a sample of nine commissural interneurones from a different population (with monosynaptic input from group I and/or II muscle afferents, and disynaptically excited from the MLF) suggest that actions of PT fibres on such interneurones are weaker because only four of them were excited by PT stimuli and at longer latencies. By demonstrating disynaptic coupling between PT neurones and commissural interneurones via reticulospinal fibres, the results provide a direct demonstration of trisynaptic coupling in the most direct pathways between PT neurones and ipsilateral motoneurones, and thereby strengthen the proposal that the double crossed pathways between PT neurones and ipsilateral motoneurones might be used to replace crossed actions of damaged PT neurones.

Same spinal interneurons mediate reflex actions of group Ib and group II afferents and crossed reticulospinal actions.

The aim of the study was to analyze interactions between neuronal networks mediating centrally initiated movements and reflex reactions evoked by peripheral afferents; specifically whether interneurons in pathways from group Ib afferents and from group II muscle afferents mediate actions of reticulospinal neurons on spinal motoneurons by contralaterally located commissural interneurons. To this end reticulospinal tract fibers were stimulated in the contralateral medial longitudinal fascicle (MLF) in chloralose-anesthetized cats in which the ipsilateral half of the spinal cord was transected rostral to the lumbosacral enlargement. In the majority of interneurons mediating reflex actions of group Ib and group II afferents, MLF stimuli evoked either excitatory or inhibitory postsynaptic potentials (EPSPs and IPSPs, respectively) or both EPSPs and IPSPs attributable to disynaptic actions by commissural interneurons. In addition, in some interneurons EPSPs were evoked at latencies compatible with monosynaptic actions of crossed axon collaterals of MLF fibers. Intracellular records from motoneurons demonstrated that both excitation and inhibition from group Ib and group II afferents are modulated by contralaterally descending reticulospinal neurons. The results lead to the conclusion that commissural interneurons activated by reticulospinal neurons affect motoneurons not only directly, but also by enhancing or weakening activation of premotor interneurons in pathways from group Ib and group II afferents. The results also show that both excitatory and inhibitory premotor interneurons are affected in this way and that commissural interneurons may assist in the selection of reflex actions of group Ib and group II afferents during centrally initiated movements.

How to enhance ipsilateral actions of pyramidal tract neurons.

We have shown previously that ipsilateral pyramidal tract (PT) neurons facilitate the actions of reticulospinal neurons on feline motoneurons (Edgley et al., 2004), which indicates that they might assist the recovery of motor functions after injuries of contralateral corticospinal neurons. Nevertheless, stimulation of ipsilateral PT fibers alone only rarely evoked any synaptic actions in motoneurons. The aim of this study was to investigate possible ways of enhancing such actions and of inducing more effective excitation and inhibition of motoneurons. The effects of stimulation of the ipsilateral PT were investigated after eliminating the spinal actions of contralateral PT fibers by hemisecting the spinal cord at a low thoracic level and were estimated from intracellular records from hindlimb motoneurons. Two measures were used to enhance PT actions. The first was to increase the probability of activation of reticulospinal neurons by mutual facilitation of actions of ipsilateral and contralateral PT neurons. The second was to enhance synaptic transmission between PT neurons and reticulospinal neurons, and in pathways between the reticulospinal neurons and motoneurons via commissural interneurons, by systemic application of a K+ channel blocker, 4-aminopyridine (4-AP). The results show that under favorable conditions, ipsilateral PT neurons may induce EPSPs and IPSPs in hindlimb motoneurons, or even action potentials, via the reticulospinal pathway. This study strengthens previous conclusions that ipsilateral PT neurons can potentially replace, at least to some extent, the actions of injured contralateral PT neurons. It also suggests that 4-AP might improve the progress of the recovery.

Positive effect of treatment with synthetic steroid hormone tibolon on intimal hyperplasia and restenosis after experimental endothelial injury of rabbit carotid artery.

BACKGROUND: Arterial intimal hyperplasia and following restenosis may be inhibited by estrogens. We investigated the effect of a synthetic steroid hormone, Tibolon: (a) on intima hyperplasia and restenosis in vivo, and (b) on production of endothelial nitric oxide synthase (eNOS), vascular endothelial growth factor (VEGF), endothelial cell proliferation and apoptosis in vitro. METHODS: Influence of Tibolon treatment (0.1 mg/kg body weight, during 3 days before and 3 weeks after the operation as a drinking solution once daily) on neointimal formation (measured by morphometry) and arterial wall damage (by qualitative histology) were investigated in vivo using an animal model of balloon injury of carotid artery. In human umbilical vein endothelial cells (HUVEC) and human microvascular endothelial cells (HMEC-1), the effect of Tibolon (0.1 microg/ml) on eNOS and VEGF was assessed by ELISA. Cell proliferation was induced by VEGF(165) and measured by BrdU incorporation assay, cell apoptosis was detected colorimetrically measuring DNA fragmentation. RESULTS: Balloon injury resulted in neointima formation and prominent damage of the carotid artery wall. Treatment with Tibolon increased luminal area, decreased intimal area and intima to media ratio, and promoted better reparation of damaged vessel wall. In vitro, Tibolon treatment did not influence the expression of eNOS protein in HUVEC as well as cell proliferation rate but reduced apoptosis of endothelial cells by about 40%. Additionally, this treatment suppressed basal and IL-1beta-stimulated synthesis of VEGF in HMEC-1. CONCLUSIONS: Tibolon treatment suppressed neointimal formation and promoted better reparation of damaged vessel wall in carotid artery after balloon injury. This positive effect seems to be associated with improved endothelial cell survival resulting possibly in increased NO production. It might be also related to the decrease of VEGF generation.

The oak processionary caterpillar as the cause of an epidemic airborne disease: survey and analysis.

BACKGROUND: Thaumetopoea processionea L. (order Lepidoptera), the oak processionary moth, is found in oak forests in most European countries. Its third to sixth larval instars are armed with poisonous hairs (setae) containing an urticating toxin (thaumetopoein) potentially harmful to humans. Because T. processionea infests trees at the edges of forests or standing alone people frequently come into contact with its setae. In the woodland bordering on the western suburbs of Vienna conditions favouring its increase have led to frequent outbreaks of lepidopterism. OBJECTIVES: To determine the incidence of lepidopterism in a suburban environment with three separate caterpillar-infested oak trees and to ascertain the frequency of the various symptoms of lepidopterism and the manner of contact with setae. METHODS: We conducted a telephone survey of all the households/institutions located within 500 m of the infested trees. To gain more information on patients' symptoms and on situations likely to lead to increased contact with setae we asked those who reported cutaneous reactions to complete a questionnaire. As part of the environmental study we described the outbreak site, examined patients and, with tape-strip samples taken from the surface of the soil, looked for setae persisting in the environment. RESULTS: Of 1025 people surveyed 57 (5.6%) reported one or more symptoms of lepidopterism: 55 (96%) reported pruritus, 54 (95%) dermatitis, eight (14%) conjunctivitis, eight (14%) pharyngitis and two (4%) respiratory distress. The questionnaire was returned by 37 (69%) of the individuals with dermatitis. Of those, 16% had reacted with weal formation, 49% with papular rash and 22% with toxic irritant dermatitis. In 13% of respondents it was not possible to define the reaction. The risk factor analysis showed that airborne contamination was the most important cause: 97% of people had frequently passed an infested tree, 57% lived near a tree (in a neighbouring garden) and 32% had a tree in their own garden. Direct contact with larvae was of minor importance (38%). In four of the tape-strip samples intact setae were identified 1 year after the infestation. CONCLUSIONS: Lepidopterism caused by T. processionea is a public health problem of increasing significance. In years with outbreaks of the pest it can reach epidemic proportions in communities located near infested trees. Contact with airborne setae was mainly responsible for the occurrence of the disease.

Inactivation of bacteriophages in water by means of non-ionizing (UV-253.7 nm) and ionizing (gamma) radiation: a comparative approach.

Thc inactivation behaviour of the bacteriophages PHI X 174 (ssDNA virus). MS2 (ssRNA virus) and B40-8 (dsDNA) toward non-ionizing (UV-253.7 nm) as well as to ionizing radiation (gamma radiation) was studied in order to evaluate their potential as viral indicators for water disinfection by irradiation. Previous findings of the high UV-253.7 nm resistance of MS2 were confirmed whereas an unexpected high sensitivity to gamma radiation compared to the two other phages was found. On the other hand, PHI X 174 revealed an enhanced UV sensitivity but a high resistance to ionizing radiation. B40-8 had an intermediate position between the other two bacteriophages relative to both types of radiation. As expected, the data of E. coli reconfirmed the unreliability of fecal indicator bacteria for the purpose of predicting responses of viruses to water treatment. In UV disinfection the influence of water matrix may be adequately controlled by considering the UV (253.7 nm) absorption of the water whereas so far no such parameter has existed for the influence of the water quality on ionizing irradiation with respect to the scavenger concentration.

UV inactivation, liquid-holding recovery, and photoreactivation of Escherichia coli O157 and other pathogenic Escherichia coli strains in water.

Drinking water, water used in food production and for irrigation, water for fish farming, waste water, surface water, and recreational water have been recently recognized as a vector for the transmission of pathogenic Escherichia coli, especially serotype O157:H7. We investigated the UV (253.7 nm) inactivation behavior and the capability of dark repair (liquid-holding recovery) and photoreactivation of seven pathogenic (including three enterohemorrhagic E. coli) strains and one nonpathogenic strain of E. coli (ATCC 11229) with respect to the use of UV light for water disinfection purposes. Because most bacteria and yeast are known to be able to repair UV damage in their nucleic acids, repair mechanisms have to be considered to ensure safe water disinfection. We found a wide divergence in the UV susceptibility within the strains tested. A 6-log reduction of bacteria that fulfills the requirement for safe water disinfection was reached for the very most susceptible strain O157:H7 (CCUG 29199) at a UV fluence of 12 J/m2, whereas for the most resistant strain, O25:K98:NM, a UV fluence of about 125 J/m2 was needed. Except for one strain (O50:H7) liquid-holding recovery did not play an important role in recovery after UV irradiation. By contrast, all strains, particularly strains O25:K98:NM, O78:K80:H12, and O157:H7 (CCUG 29193), demonstrated photorepair ability. For a 6-log reduction of these strains, a UV fluence (253.7 nm) up to 300 J/m2 is required. The results reveal that the minimum fluence of 400 J/m2 demanded in the Austrian standard for water disinfection is sufficient to inactivate pathogenic E. coli. A fluence of 160 J/m2 (recommendation in Norway) or 250 J/m2 (recommendation in Switzerland) cannot be regarded as safe in that respect.

Measurement of UV radiation using suspensions of microorganisms.

The measurement of solar UV radiation is usually performed using physical devices like photodiodes or photomultipliers or with chemical substances (actinometry). The application of biological material such as microorganisms for this purpose has gained increasing importance in the last few years. The microorganisms may be dried and spread on a flat surface or they may be in aqueous suspensions contained in UV-transparent vessels. If the measurements are done on flat surfaces, the irradiance weighted by the action spectrum of the dried microorganism used is the result of the measurement. If aqueous suspensions of microorganisms are used, contained for instance in spherical vessels, the fluence weighted by the action spectrum of the microorganisms in the aqueous suspension is the result. A problem of this method of measurement can be that inside the vessel the distribution of UV radiation is usually not homogeneous, causing distributions of fluences among the irradiated microorganisms, which may result in variation of the results depending on the mixing characteristics of the suspension during irradiation.

Increased inactivation of Saccharomyces cerevisiae by protraction of UV irradiation.

The principle of equi-effectivity of the product of intensity and exposure time (principle of Bunsen-Roscoe) of UV irradiation has been assumed to be valid for the inactivation of microorganisms in general. Earlier studies claimed higher survival of Escherichia coli B/r with fractionated irradiation compared with single-exposure survival. However, data on the inactivation effect of protraction of UV irradiation are not available. By means of a specially designed UV irradiation apparatus which secured absolute UV dose measurements throughout the experiments, the effects of variation of UV irradiation intensities (253.7 nm) and exposure times were tested on the inactivation of a bacterial virus (Staphylococcus aureus phage A994), a vegetative bacterial strain (E. coli ATCC 25922), and bacterial spores (Bacillus subtilis ATCC 6633) as well as three haploid laboratory strains (RC43a, YNN281, and YNN282) and two diploid strains (commercial bakery yeast strain and laboratory strain YNN281 x YNN282) or yeast (Saccharomyces cerevisiae) and spores of the latter diploid yeast strain. Each test organism was exposed to three UV intensities (0.02, 0.2, and 2 W/m2), with corresponding exposure times resulting in three dose levels for each intensity. Differences in inactivation rates were tested by analyses of variance and Newman-Keuls tests. Virus and bacteria showed no differences in inactivation rates by variation of intensities and exposure times within selected UV doses; hence, the principle of Bunsen-Roscoe could not be rejected for these strains. However, in the eukaryotic test strains of S. cerevisiae longer exposure times with lower intensities led to enhanced inactivation in both haploid and diploid strains, with a more pronounced effect in the diploid yeast strains, whereas in yeast spores in this dose rate effect could not be observed.

[Human behavior in the solar radiation field with reference to ultraviolet exposure]. / Das Verhalten des Menschen im solaren Strahlungsfeld im Hinblick auf seine UV-Exposition.

There is a causal relation between solar ultraviolet radiation and skin cancer. For epidemiological investigations, quantification of the UV exposure is essential. To set up a risk assessment for the whole population, a representative survey was performed in Austria. The questionnaire refers to three sectors of everyday life: work, recreation and holidays; in addition the use of solaria is asked about for a further investigation. The UV exposure caused by humans' behaviour in the field of solar radiation was analysed from various demographic aspects. For some subpopulations the UV exposure sustained during work, recreation and holidays was compared. Groups with high occupational UV exposure show a weaker tendency to stay outdoors during leisure time and holidays than groups characterized by high UV exposure in their leisure time, who also prefer sun-intensive activities during holidays.

Trend analysis of solar ultraviolet exposure of the Austrian population caused by holiday patterns since 1969.

Over the last few decades, a steady increase in the incidence of skin cancer has been observed. Solar ultraviolet (UV) exposure is undoubtedly one of the primary causes, and of these, holiday patterns are an intrinsic component. To assess solar UV exposure for this trend analysis, changes in holiday patterns were investigated. For Austrians, a preponderance of holidays are spent at the Mediterranean Sea. These holidays represent not only a potentially high UV exposure because of location but also because sun-intensive activities are preferred. The analysis shows an increase in solar UV exposure from all holidays of 1.6% per year. About 22% of the population spends holidays at the Mediterranean Sea, and UV exposure of the total population from these holidays has increased annually by about 3.3%. Because of the latency period, it is not possible to draw conclusions from the increase in UV exposure and the temporal trends in skin cancer. A longer time series would be necessary to do this. A direct comparison of data from other countries reporting increases in skin cancer does not seem appropriate, as levels of UV exposure vary greatly and such factors as genetic susceptibility are involved.

[UV-inactivation of microorganisms in water]. / UV-Inaktivierung von Mikroorganismen in Wasser.

UV-Inactivation of Escherichia coli, Bacillus subtilis spores, Staphylococcus-Phage A 994, Poliovirus type Mahoney and Rotavirus SA 11 was tested under controlled physical conditions. B. subtilis-spores were found to be the most resistant of these microorganisms, followed by Rotavirus, Bacteriophage and Poliovirus. E. coli required the lowest irradiation dose for inactivation. Causes and meaning of these dose-survival-reactions are discussed.

[Construction of tissue compensating filters for extended areas in Hodgkin-irradiation on the basis of moiré topography (author's transl)]. / Herstellung von Gewebeausgleichsfiltern für gorsse Flächen bei Hodgkin-Bestrahlung mit Hilfe der Moiré-Topographie

In order to obtain a homogeneous distribution of the dose over extended fields in radiotherapy of Hodgkin's disease, for an exact dosage within the irradiated region it is necessary to consider the topography of the patient. The skin protective build-up effect due to the use of bolus materials would be lost with telecobalt irradiation, and therefore tissue compensating filters with a large surface are to be preferred. A simple method of making such filters by means of the moirétopography is described. The way to obtain a compensation filter for extended areas is demonstrated by an example showing how to construct it from lead plates correct in scale and layerwise.