Restoration of the Dopamine Transporter through Cell Therapy Improves Dyskinesia in a Rat Model of Parkinson's Disease.

The dyskinesia of Parkinson's Disease is most likely due to excess levels of dopamine in the striatum. The mechanism may be due to aberrant synthesis but also, a deficiency or absence of the Dopamine Transporter. In this study we have examined the proposition that reinstating Dopamine Transporter expression in the striatum would reduce dyskinesia. We transplanted c17.2 cells that stably expressed the Dopamine Transporter into dyskinetic rats. There was a reduction in dyskinesia in rats that received grafts expressing the Dopamine Transporter. Strategies designed to increase Dopamine Transporter in the striatum may be useful in treating the dyskinesia associated with human Parkinson's Disease.

Distribution of orexin-1 receptor-green fluorescent protein- (OX1-GFP) expressing neurons in the mouse brain stem and pons: Co-localization with tyrosine hydroxylase and neuronal nitric oxide synthase.

We used a reporter mouse line in which green fluorescent protein (GFP) was inserted into the orexin-1 receptor (OX1) locus to systematically map the neuroanatomical distribution of the OX1 receptor in the mouse brainstem and pons. Here, we show that the OX1 receptor is expressed in a select subset of medullary and pontine nuclei. In the medulla, we observed OX1-GFP expression in the cuneate, gracile, dorsal motor nucleus of the vagus (10N), nucleus of the solitary tract and medullary raphe areas. In the pons, the greatest expression was found in the locus coeruleus (LC) and dorsal raphe nucleus (DRN). High to moderate expression was found in the pedunculopontine tegmental nucleus (PPTg), laterodorsal tegmental nucleus, A5 noradrenergic cell group (A5) and the periaqueductal gray. Double-labeling with neuronal nitric oxide synthase (NOS1) revealed extensive co-localization in cell bodies and fibers of the 10N, A5 cell group and the PPTg. Double-staining with tyrosine hydroxylase revealed extensive co-expression in the LC, DRN and the lateral paragigantocellularis cell group in the ventral medulla. Our findings faithfully recapitulate the findings of OX1 mRNA expression previously reported. This is the first study to systematically map the neuroanatomical distribution of OX1 receptors within the mouse hindbrain and suggest that this OX1-GFP transgenic reporter mouse line might be a useful tool with which to study the neuroanatomy and physiology of OX1 receptor-expressing cells.

Measurements of neutron radiation in aircraft.

Radiation environment is a complex mixture of charged particles of the solar and galactic origin, as well as of secondary particles created in an interaction of galactic cosmic particles with the nuclei of the Earth's atmosphere. A radiation field at aircraft altitude consists of different types of particles, mainly photons, electrons, positrons and neutrons, with a large energy range. In order to measure a neutron component of the cosmic radiation, we investigated a few combinations of a track etch detector (CR-39, LR-115) with a plastic converter or boron foil. Detector calibration was performed on neutrons coming from the nuclear reactor, as well as in the CERN-EU high-energy Reference Field (CERF) facility. From November 2007 to September 2008, the neutron dose equivalent was measured by the track detectors during five aircraft flights, in the north geographical latitude from 21° to 58°; the respective average dose rate, determined by using the D-4 detector (CR-39/B), was H(n)=5.9 µSv/h. The photon dose rate, measured by the electronic dosimeter RAD-60 SE, had the average value of H(f)=1.4 µSv/h.

Allergenicity and immunogenicity of the major mugwort pollen allergen Art v 1 chemically modified by acetylation.

BACKGROUND: Treating allergies with modified allergens is an approach to make the treatment safer and more efficient. Art v 1 is the most prominent allergen of mugwort pollen and a significant cause of hayfever around Europe. The aim of this study was to reduce the allergenicity of Art v 1 by acetylation, and to investigate the capacity of the modified protein to generate blocking antibodies. METHODS: The reduction of allergenicity of Art v 1 following acetylation was monitored by immunoblot, ELISA inhibition using a pool of sera from mugwort pollen allergic patients, basophil activation assay and by skin prick testing of mugwort-allergic patients. Rabbits were immunized against Art v 1 and acetylated Art v 1 (acArt v 1) and the rabbit antisera were tested for their capacity to block human IgE binding in ELISA. Human T cell proliferation against Art v 1 and acArt v 1 was examined in peripheral blood mononuclear cells (PBMCs) of mugwort pollen allergic patients and cytokine release in PBMC cultures was monitored. RESULTS: Acetylation of Art v 1 gave a derivative of reduced allergenicity in the in vitro and ex vivo tests applied. The skin test reactivity to acArt v 1 was significantly reduced in 19 patients when compared with the reactivity to Art v 1. Rabbit antibodies to acArt v 1 and Art v 1 showed similar capacity to block human IgE binding to Art v 1 in inhibition ELISA. Both proteins were able to induce proliferation of PBMCs and CD3/CD4(+) cells of mugwort-allergic patients. Release of IL-5 was significantly reduced in cultures stimulated with acArt v 1. CONCLUSIONS: Art v 1 modified by acetylation had a significantly reduced allergenicity in vitro and in vivo, while its immunogenicity was retained. Modification of allergens by acetylation could be a new strategy for allergen-specific immunotherapy.

Radon anomaly in soil gas as an earthquake precursor.

The mechanical processes of earthquake preparation are always accompanied by deformations; afterwards, the complex short- or long-term precursory phenomena can appear. Anomalies of radon concentrations in soil gas are registered a few weeks or months before many earthquakes. Radon concentrations in soil gas were continuously measured by the LR-115 nuclear track detectors at site A (Osijek) during a 4-year period, as well as by the Barasol semiconductor detector at site B (Kasina) during 2 years. We investigated the influence of the meteorological parameters on the temporal radon variations, and we determined the equation of the multiple regression that enabled the reduction (deconvolution) of the radon variation caused by the barometric pressure, rainfall and temperature. The pre-earthquake radon anomalies at site A indicated 46% of the seismic events, on criterion M>or=3, R<200 km, and 21% at site B. Empirical equations between earthquake magnitude, epicenter distance and precursor time enabled estimation or prediction of an earthquake that will rise at the epicenter distance R from the monitoring site in expecting precursor time T.

Alterations in the proportions of skeletal muscle proteins following a unilateral lesion to the substantia nigra pars compacta of rats.

It is well established that mammalian skeletal muscles exhibit a considerable degree of plasticity and one of the main determining factors of this plasticity is the activity pattern and duration of motoneurone discharge. Lesions to the right substantia nigra pars compacta (SNpc) of six adult rats were made to determine whether altered output from the SNpc ultimately leads to a change in the expression of proteins in contralateral skeletal muscles. After 4 months, altered motor performance was identified by the administration of amphetamine. After 7 months, 30-70% of dopaminergic cells in the SNpc had been destroyed. The protein content of muscles was then quantified from densitometric scans of gels, and expressed as a % of the amount of actin (the protein used as a reference in this study). The lesion affected the expression of different protein isoforms in the fast- and slow-twitch muscles. In slow-twitch soleus muscles, the lesion decreased the proportion of alpha-tropomyosin and increased the proportion of beta-tropomyosin. In the fast-twitch extensor digitorum longus muscles, the lesion increased the proportion of the fast isoform of troponin-T1f, and decreased the proportions of the two isoforms of myosin light chain. This study establishes a connection between the chronic effects of a lesion to the SNpc, with a loss of dopaminergic neurones, impaired motor performance, and altered expression of proteins in skeletal muscle. The implication of these results is that the altered motor function observed in Parkinson's disease may be associated with alterations to the expression of skeletal muscle proteins.

Radon in the spas of Croatia.

Radon concentrations in air and geothermal water of the spa pools in Croatia were measured and the average values of 40.3 and 4.5 kBq/m3 were obtained, respectively. Great difference between radon concentrations in pool and spring water was considered as a result of mixing normal and geothermal water in the pool as well as the radon decay. Estimation of an effective dose, received by the personnel in the Bizovac spa, gave the value of 0.27 mSv/y. At the location Stubica, the transfer factor of the radon for air and thermal water in the pool was calculated, and the value of 4.9+/-0.7 x 10(-3) was obtained.

Haloperidol treatment reverses behavioural and anatomical changes in cocaine-dependent mice.

Abnormal dopamine (DA) transmission occurs in many pathological conditions, including drug addiction. Previously, we showed DA D2 receptor (D2R) activation results in pruning of the axonal arbour of DA neurones that innervate the dorsal striatum. Thus, we hypothesised that long-term D2R stimulation through drugs of addiction should cause arbour pruning of neurones that innervate the ventral striatum and thus reduce DA release and contribute to craving. If so, D2R blockade should return these arbours to normal size and may overcome craving. We show that long-term treatment with a D2R antagonist (haloperidol) reverses behavioural and anatomical effects of cocaine dependence in mice, including relapse. This change in arbour size reflects new synapse formation and our data suggest this must occur in the presence of increased DA activity to reverse cocaine-seeking behaviour. These findings hold significant implications for the understanding and treatment of cocaine addiction.

Timecourse of striatal re-innervation following lesions of dopaminergic SNpc neurons of the rat.

Previously we described the extent of sprouting that axons of the rat substantia nigra pars compacta (SNpc) undergo to grow new synapses and re-innervate the dorsal striatum 16 weeks after partial lesions. Here we provide insights into the timing of events related to the re-innervation of the dorsal striatum by regenerating dopaminergic nigrostriatal axons over a 104-week period after partial SNpc lesioning. Density of dopamine transporter and tyrosine hydroxylase immunoreactive axonal varicosities (terminals) decreased up to 80% 4 weeks after lesioning but returned to normal by 16 weeks, unless SNpc lesions were greater than 75%. Neuronal tracer injections into the SNpc revealed a 119% increase in axon fibres (4 mm rostral to the SNpc) along the medial forebrain bundle 4 weeks after lesioning. SNpc cells underwent phenotypic changes. Four weeks after lesioning the proportion of SNpc neurons that expressed tyrosine hydroxylase fell from 90% to 38% but returned to 78% by 32 weeks. We discuss these phenotype changes in the context of neurogenesis. Significant reductions in dopamine levels in rats with medium (30-75%) lesions returned to normal by 16 weeks whereas recovery was not observed if lesions were larger than 75%. Finally, rotational behaviour of animals in response to amphetamine was examined. The clear rightward turning bias observed after 2 weeks recovered by 16 weeks in animals with medium (30-75%) lesions but was still present when lesions were larger. These studies provide insights into the processes that regulate sprouting responses in the central nervous system following injury.

Changes in function and ultrastructure of striatal dopaminergic terminals that regenerate following partial lesions of the SNpc.

Following partial substantia nigra lesions, remaining dopaminergic neurones sprout, returning terminal density in the dorsal striatum to normal by 16 weeks. This suggests regeneration and maintenance of terminal density is regulated to release appropriate levels of dopamine. This study examined the structure and function of these reinnervated terminals, defining characteristics of dopamine uptake and release, density and affinity of the dopamine transporter (DAT) and ultrastructural morphology of dopamine terminals in the reinnervated dorsal striatum. Finally, rotational behaviour of animals in response to amphetamine was examined 4 and 16 weeks after substantia nigra pars compacta (SNpc) lesions. Dopamine transport was markedly reduced 16 weeks after lesioning along with reduced density and affinity of DAT. Rate of dopamine release and peak concentration, measured electrochemically, was similar in lesioned and control animals, while clearance was prolonged after lesioning. Ultrastructurally, terminals after lesioning were morphologically distinct, having increased bouton size, vesicle number and mitochondria, and more proximal contacts on post-synaptic cells. After 4 weeks, tendency to rotate in response to amphetamine was proportional to lesion size. By 16 weeks, rotational behaviour returned to near normal in animals where lesions were less than 70%, although some animals demonstrated unusual rotational patterns at the beginning and end of the amphetamine effect. Together, these changes indicate that sprouted terminals are well compensated for dopamine release but that transport mechanisms are functionally impaired. We discuss these results in terms of implications for dyskinesia and other behavioural states.

D2 Dopamine receptor blockade results in sprouting of DA axons in the intact animal but prevents sprouting following nigral lesions.

Recently it was demonstrated that sprouting of dopaminergic neurons and a microglial and astrocyte response follows both partial lesions of the substantia nigra pars compacta and blockade of the D2 dopamine receptor. We therefore studied the effects of the combination of these two treatments (lesioning and D2 dopamine receptor blockade). Haloperidol administration caused a 57% increase in dopaminergic terminal tree size (measured as terminal density per substantia nigra pars compacta neuron) and an increase of glia in the striatum. Following small to medium nigral lesions (less than 60%), terminal tree size increased by 51% on average and returned density of dopaminergic terminals to normal. In contrast, administration of haloperidol for 16 weeks following lesioning resulted in reduced dopaminergic terminal density and terminal tree size (13%), consistent with absent or impaired sprouting. Glial cell numbers increased but were less than with lesions alone. When haloperidol was administered after the striatum had been reinnervated through sprouting (16-32 weeks after lesioning), terminal tree size increased up to 150%, similar to the effect of haloperidol in normal animals. By examining the effect of administering haloperidol at varying times following a lesion, we concluded that a switch in the effect of D2 dopamine receptor blockade occurred after dopaminergic synapses began to form in the striatum. We postulate that when synapses are present, D2 dopamine receptor blockade results in increased terminal density, whereas prior to synapse formation D2 dopamine receptor blockade causes attenuation of a sprouting response. We speculate that D2 dopamine receptors located on growth cones 'push' neurites toward their targets, and blockade of these receptors could lead to attenuation of sprouting.

Effects of long-term treatment with dopamine receptor agonists and antagonists on terminal arbor size.

This study demonstrates that pharmacological manipulation of the dopamine (DA) receptors can modulate the size of the axonal tree of substantia nigra pars compacta (SNpc) neurons in mice. Pharmacological blockade or genetic ablation of the D2 receptor (D2R) resulted in sprouting of DA SNpc neurons whereas treatment with a D2 agonist resulted in pruning of the terminal arbor of these neurons. Agents such as cocaine, that indirectly stimulate D2R, also resulted in reduced terminal arbor. Specific D1 agonists or antagonists had no effect on the density of DA terminals in the striatum. We conclude that the D2 receptor has a central role in regulating the size of the terminal arbor of nigrostriatal neurons. These findings have implications relating to the use of dopaminergic agonists in the management of Parkinson's disease and in controlling plasticity following injury, loss or transplantation of DA neurons.

Axonal sprouting following lesions of the rat substantia nigra.

Parkinson's disease is characterized by the progressive loss of dopaminergic neurons in the substantia nigra pars compacta. Symptoms do not appear until most nigral neurons are lost, implying that compensatory mechanisms are present. Sprouting has been proposed as one of these mechanisms. This study quantified the extent of compensatory axonal sprouting following injury of dopaminergic neurons within the substantia nigra pars compacta. Specifically, the extent of the axonal arbour and axonal varicosity morphology was measured after partial destruction (with 6-hydroxydopamine) of the substantia nigra of the adult male rat. Four months later, the substantia nigra was injected with the anterograde neuronal tracer dextran-biotin to trace the full extent of individual axons. An unbiased estimate of neuron number was performed in each animal. This demonstrated nigral neuronal loss ranging from 10 to 90% on the side that received the injection whilst a 7% reduction was observed in the side contralateral to the lesion. Coincident with this loss, some nigral neurons lose tyrosine hydroxylase expression. Vigorous axonal sprouting was observed in the terminal arbours of lesioned animals and was associated with an increased axonal varicosity size. Axonal varicosities and branching points were primarily confined to the dorsal 1.5mm of the caudate-putamen, an area predominantly innervated by nigral neurons. It appears that dopaminergic neurons were responsible for this sprouting because the density of dopamine transporter immunoreactive varicosities in the caudate-putamen was maintained until about a 70% loss of neurons. It was concluded that substantial compensation in the form of sprouting and new dopaminergic synapse formation occurs following lesions of the substantia nigra pars compacta.

[Psychosocial research during the war in Sarajevo]. / Psihosocijalna istrazivanja u toku rata u Sarajevu.

War in Bosnia and Herzegovina has caused many psychic and social breakdowns. The consequences on mental health of the war which caused stress are of importance, as well as influences on psychic functioning of individuals are caused by changes in social structure of population and economic potential of the society. Project "Psycho-social aspects of war in BiH" carried out within the frame of the Academy of Sciences and Arts of Bosnia and Herzegovina and the Department of Psychiatry of Clinical Center of Sarajevo University. In this article are given the results of the Project, but only partially. The investigations showed that the number of patients visiting hospitals during the war was greatly increased in the field of stress reactions and reactive psychoses. But incidence and prevalence of alcohol psychoses decreased. Findings are the same for out patient clinics. The field investigation on the free territories of Sarajevo s communities shows enormous increase of mental disorders among the citizens: neurotic over 40%, psychotic about 20% and that is, together, more than 60% of the population of the town Sarajevo were disturbed at that time. Among the children and adolescents there was an increase of neurotic and psychotic disorders in the very beginning of the first year of the war, and decrease of the same diagnoses during the second year. This might be explained by particular adaptation of the youngsters to war conditions. When we are talking about invalidity of neurological and psychiatric disorders, the investigations showed that illness is the mostly caused by invalidity (85.1%) among the global invalidity during the war in Sarajevo. Injuries before the war were at 3%, during the war are 11% of cases. But, all those shows temporary, because war caused invalidity more and we are expecting to registration later. Our investigation among the refugee camps and population in Sarajevo shows that "life equipment" among the displaced persons was lower than domestic people. That shows that after the phase of surviving this part of the population was at risk of many psycho-social problems. Also, our investigation shows that very low socio-economical level of inhabitants of Sarajevo leads to the potential of absolutely poverty. The indicators of this trend are: low level of education, very low life standard, unemployment, bad health conditions etc. Among refugees all those indicators are worse. Criminality in Sarajevo during the war has been increased, particularly among adolescents. One fifth of contents of daily newspaper "Oslobodenje" has been during the war oriented to the health system problems.