Toxicity and elemental composition of particulate matter from outdoor and indoor air of elementary schools in Munich, Germany.

UNLABELLED: Outdoor particulate matter (PM(10)) is associated with detrimental health effects. However, individual PM(10) exposure occurs mostly indoors. We therefore compared the toxic effects of classroom, outdoor, and residential PM(10). Indoor and outdoor PM(10) was collected from six schools in Munich during teaching hours and in six homes. Particles were analyzed by scanning electron microscopy and X-ray spectroscopy (EDX). Toxicity was evaluated in human primary keratinocytes, lung epithelial cells and after metabolic activation by several human cytochromes P450. We found that PM(10) concentrations during teaching hours were 5.6-times higher than outdoors (117 ± 48 µg/m(3) vs. 21 ± 15 µg/m(3), P < 0.001). Compared to outdoors, indoor PM contained more silicate (36% of particle number), organic (29%, probably originating from human skin), and Ca-carbonate particles (12%, probably originating from paper). Outdoor PM contained more Ca-sulfate particles (38%). Indoor PM at 6 µg/cm(2) (10 µg/ml) caused toxicity in keratinocytes and in cells expressing CYP2B6 and CYP3A4. Toxicity by CYP2B6 was abolished with the reactive oxygen species scavenger N-acetylcysteine. We concluded that outdoor PM(10) and indoor PM(10) from homes were devoid of toxicity. Indoor PM(10) was elevated, chemically different and toxicologically more active than outdoor PM(10). Whether the effects translate into a significant health risk needs to be determined. Until then, we suggest better ventilation as a sensible option. PRACTICAL IMPLICATIONS: Indoor air PM(10) on an equal weight base is toxicologically more active than outdoor PM(10). In addition, indoor PM(10) concentrations are about six times higher than outdoor air. Thus, ventilation of classrooms with outdoor air will improve air quality and is likely to provide a health benefit. It is also easier than cleaning PM(10) from indoor air, which has proven to be tedious.

Can immunoglobulin E-measurement replace challenge tests in allergic rhinoconjunctivits to grass pollen?

BACKGROUND: There is minimal data available concerning the dose-response relationship between allergen exposure and clinical reactivity for outdoor aeroallergens, such as timothy grass pollen. Timothy pollen-specific IgE (sIgE) determinations might assist in predicting the clinical reactivity in patients with allergic rhinoconjunctivitis (ARC). METHODS: Allergen-sIgE antibody levels of timothy grass pollen were correlated with individual threshold doses eliciting allergic reactions in skin prick test (SPT), conjunctival (CPT) and nasal (NPT) provocation tests in patients suffering from pollen-induced rhinoconjunctivitis and healthy controls. RESULTS: One hundred and four patients with ARC (median age: 27 years; range: 18-64; females: 58%) and 36 controls (25 years (22-77); females: 70%) were included in the study. Ninety-six percent of the patients showed a positive reaction in the nasal and 57% showed a positive reaction in the conjunctival provocation. With regarding to titrated SPT, 98% of the patients showed a positive skin test reaction; correlating with the level of sIgE for timothy (P < 0.001). For both provocation protocols, there was no correlation between the provocation concentration at the reaction and the level of sIgE for timothy. The ratio of sIgE/total IgE correlated with the dilution level of SPT (P < 0.002) and CPT (P < 0.01), respectively. CONCLUSION AND CLINICAL RELEVANCE: A dose-response relationship between the levels of sIgE and clinical outcome of timothy allergen exposure could not be established. Although IgE-determination remains an important key element in allergy diagnosis, provocation tests are procedures of choice if the clinical relevance of an allergen has to be confirmed.

The allergen Bet v 1 in fractions of ambient air deviates from birch pollen counts.

BACKGROUND: Proof is lacking that pollen count is representative for allergen exposure, also because allergens were found in nonpollen-bearing fractions of ambient air. OBJECTIVE: We monitored simultaneously birch pollen and the major birch pollen allergen Bet v 1 in different size fractions of ambient air from 2004 till 2007 in Munich, Germany. METHODS: Air was sampled with a ChemVol high-volume cascade impactor equipped with stages for particulate matter (PM)>10 microm, 10 microm>PM>2.5 microm, and 2.5 microm>PM>0.12 microm. Allergen was determined with a Bet v 1-specific ELISA. Pollen count was assessed with a Burkard pollen trap. We also measured the development of allergen in pollen during ripening. RESULTS: About 93 +/- 3% of Bet v 1 was found in the PM > 10 microm fraction, the fraction containing birch pollen. We did not measure any Bet v 1 in 2.5 microm > PM > 0.12 microm. Either in Munich no allergen was in this fraction or the allergen was absorbed to diesel soot particles that also deposit in this fraction. Pollen released 115% more Bet v 1 in 2007 than in 2004. Also within 1 year, the release of allergen from the same amount of pollen varied more than 10-fold between different days. This difference was explained by a rapidly increasing expression of Bet v 1 in pollen in the week just before pollination. Depending on the day the pollen is released during ripening, its potency varies. CONCLUSION: In general, pollen count and allergen in ambient air follow the same temporal trends. However, because a 10-fold difference can exist in allergen potency of birch pollen, symptoms might be difficult to correlate with pollen counts, but perhaps better with allergen exposure.

Influence of acupuncture on type I hypersensitivity itch and the wheal and flare response in adults with atopic eczema - a blinded, randomized, placebo-controlled, crossover trial.

BACKGROUND: Itch is a major symptom of allergic skin disease. Acupuncture has been shown to exhibit a significant effect on histamine-induced itch in healthy volunteers. We investigated the effect of acupuncture on type I hypersensitivity itch and skin reaction in a double-blind, randomized, placebo-controlled, crossover trial. METHODS: An allergen stimulus (house dust mite or grass pollen skin prick) was applied to 30 patients with atopic eczema before (direct effect) and after (preventive effect) two experimental approaches or control observation: acupuncture at points Quchi and Xuehai [verum acupuncture (VA), dominant side], 'placebo-point' acupuncture (PA, dominant side), no acupuncture (NA). Itch intensity was recorded on a visual analogue scale. After 10 min, wheal and flare size and skin perfusion (via LASER-Doppler) were measured at the stimulus site, and the validated Eppendorf Itch Questionnaire (EIQ) was answered. RESULTS: Mean itch intensity was significantly lower in VA (35.7 +/- 6.4) compared to NA (45.9 +/- 7.8) and PA (40.4 +/- 5.8) regarding the direct effect; and significantly lower in VA (34.3 +/- 7.1) and PA (37.8 +/- 5.6) compared to NA (44.6 +/- 6.2) regarding the preventive effect. In the preventive approach, mean wheal and flare size were significantly smaller in VA (0.38 +/- 0.12 cm(2)/8.1 +/- 2.0 cm(2)) compared to PA (0.54 +/- 0.13 cm(2)/13.5 +/- 2.8 cm(2)) and NA (0.73 +/- 0.28 cm(2)/15.1 +/- 4.1 cm(2)), and mean perfusion in VA (72.4 +/- 10.7) compared to NA (84.1 +/- 10.7). Mean EIQ ratings were significantly lower in VA compared to NA and PA in the treatment approach; and significantly lower in VA and PA compared to NA in the preventive approach. CONCLUSIONS: Acupuncture at the correct points showed a significant reduction in type I hypersensitivity itch in patients with atopic eczema. With time the preventive point-specific effect diminished with regard to subjective itch sensation, whereas it increased in suppressing skin-prick reactions.

Gender difference, sex hormones, and immediate type hypersensitivity reactions.

Gender differences in the development and prevalence of human diseases have long been recognized. Immense interest grows in the understanding of the role of sex hormones in the homeostasis of immunity. Asthma predominates in boys before puberty and this gender preference reverses after puberty and in adulthood, when adult women tend to have a more severe disease, often recalcitrant to treatment. Atopic eczema in preschool children shows insignificant gender difference or male preponderance in different studies, with more adult females suffering from atopic eczema. The limited data on the prevalence of immediate hypersensitivity to hymenoptera venom show controversial results. Discrepancy exists regarding the gender difference in food allergy, with females reporting significantly more allergic reactions in questionnaire studies. In general, adverse reactions to nonionic iodinated radiocontrast media are more commonly observed in females. The course of allergic diseases varies unpredictably during pregnancy, whereas hormone replacement therapy in postmenopausal women usually has a favorable influence on the course of asthma. Experiments in rodents confirm an effect of estrogens on mast cell activation and allergic sensitization, while progesterone is shown to suppress histamine release but potentiate IgE induction. Dehydroepiandrosterone may antagonize the production of Th2 cytokines but the effect of testosterone and the other androgens remains less defined. Actual data from human studies are lacking.

[Influence of environmental factors on allergy development]. / Einfluss von Umweltfaktoren auf die Allergieentstehung.

The increase in allergic diseases is inter alia explained by the adjuvant effect of environmental pollutants: (1) The interaction between traffic-related airborne particles and pollen grains in the atmosphere may lead to agglomeration of particles on the surface of allergen carriers inducing their activation and to modulation of allergen release, generation of allergenic aerosols and adsorption of pollen proteins to airborne particles. (2) Anthropogenic air pollutants enhance the release of pollen-associated lipid mediators (PALMs) from pollen grains, substances with proinflammatory and immune modulating effects, which can lead to enhancement of allergic symptoms and maintenance of disease. (3) Air pollutants, such as NO(2), ozone, secondhand tobacco smoke, fine and ultrafine particles play an important role as adjuvants and trigger factors for allergic disease development as well as for elicitation and aggravation of allergic symptoms. (4) Polymorphisms in phase II drug-metabolizing enzymes can modulate susceptibility to the adjuvant effects of anthropogenic air pollutants on the IgE-mediated immune response. This highlights gene-environment interactions, which play an important role in the manifestation of allergic diseases.

Leukotriene B4 receptor inhibitor LY293111 induces cell cycle arrest and apoptosis in human anaplastic large-cell lymphoma cells via JNK phosphorylation.

Anaplastic large-cell lymphoma (ALCL) is a heterogeneous lymphoma category in which a subset of cases carry the t(2;5)(p23;q35) or variant translocations resulting in overexpression of anaplastic lymphoma kinase (ALK). LY293111 (2-[2-propyl-3-[3-[2-ethyl-4-(4-fluorophenyl)-5-hydroxyphenoxy]-propoxy]-phenoxy] benzoic acid sodium salt) is a leukotriene B4 receptor antagonist, which was found to be safe and tolerable in Phase I clinical trials. In this study, we investigated the potential therapeutic effects and mechanisms of action of LY293111 in ALCL cell lines. LY293111 inhibited proliferation of both ALK(+) and ALK(-) ALCL cell in a dose-dependent fashion and induced complete G(1)-S cell cycle arrest, which was accompanied by upregulation of p27 and downregulation of cyclin E. Pretreatment with LY293111 for 4 h resulted in profound inhibition of serum-induced phosphorylation of extracellular-regulated kinases-1 and 2 and Akt and a concomitant increase in the phosphorylation of the stress-activated kinase c-jun N-terminal kinases (JNK). Simultaneously, LY293111 induced caspase-dependent apoptosis via activation of the intrinsic pathway, including early loss of mitochondrial inner transmembrane potential and the production of reactive oxygen species (ROS), cleavage of caspases-9, -3, poly ADP-ribose polymerase (PARP) and X-linked inhibitor of apoptosis. The phospho-JNK inhibitor SP600125 partially protected Sup-M2 cells from LY293111-induced apoptosis, PARP cleavage and ROS generation, suggesting a role for JNK in LY293111-induced cell death. These results warrant further studies of LY293111 in ALCL.

Regulation and targeting of antiapoptotic XIAP in acute myeloid leukemia.

XIAP is a member of the inhibitors-of-apoptosis family of proteins, which inhibit caspases and block cell death, with prognostic importance in AML. Here we demonstrate that cytokines regulate the expression of XIAP in leukemic cell lines and primary AML blasts. Inhibition of phosphatidylinositol-3 kinase (PI3K) with LY294002 and of the mitogen-activated protein kinase (MAPK) cascade by PD98059 resulted in decreased XIAP levels (34+/-8.7 and 23+/-5.7%, respectively). We then generated OCI-AML3 cells with constitutively phosphorylated Akt (p473-Akt) by retroviral gene transfer. Neither these nor Akt inhibitor-treated OCI-AML3 cells showed changes in XIAP levels, suggesting that XIAP expression is regulated by PI3K downstream effectors other than Akt. The induction of XIAP expression by cytokines through PI3K/MAPK pathways is consistent with its role in cell survival. Exposure of leukemic cells to chemotherapeutic agents decreased XIAP protein levels by caspase-dependent XIAP cleavage. Targeting XIAP by XIAP antisense oligonucleotide resulted in downregulation of XIAP, activation of caspases and cell death, and sensitized HL-60 cells to Ara-C. Our results suggest that XIAP is regulated by cytokines through PI3K, and to a lesser degree through MAPK pathways. Selective downregulation of XIAP expression might be of therapeutic benefit to leukemic patients.

Increased expression of adhesive proteins on leukocytes by TNF alpha.

To further elucidate the role of tumor necrosis factor alpha (TNF alpha) in cell adhesion, we investigated the effect of TNF alpha on the expression of surface adhesive protein. After treatment with TNF alpha for 1 h, the increased expression of surface adhesive proteins (beta subunit) was observed on granulocytes but not on monocytes or lymphocytes. The expression of Mac-1 (3,4-fold increase) was consistently enhanced more than p 150,95 (1.4-fold increase) and LFA-1 expression was unchanged. Dose-response and time course studies indicated a parallel relationship between TNF alpha-increased expression of surface adhesive proteins and TNF alpha-induced granulocyte adhesion. The anti-inflammatory drug Dexamethasone suppressed both TNF alpha-induced granulocyte adhesion and TNF alpha-induced expression of surface adhesive proteins. The inhibition of granulocyte adhesion correlated with the reduction of surface adhesive protein expression. The data suggest that one contributing factor in the mechanism by which Dexamethasone inhibited TNF alpha-induced granulocyte adhesion may be diminished expression of surface adhesive proteins.

NADPH-diaphorase in the central nervous system of the larval lamprey (Lampetra planeri).

The distribution of nitric oxide synthase (NOS) in the lamprey brain was studied by using reduced nicotinamide-adenine-dinucleotide phosphate (NADPH)-diaphorase histochemistry to further elucidate the evolution of neurons synthesizing nitric oxide. Intense labeling of fibers and/or neurons was found in portions of the lamprey central nervous system, such as the olfactory system, the pineal organ, the habenular region, the nervus stato-acousticus (N. VIII), the brainstem, and the spinal cord, and also in the adenohypophysis. Labeled giant cells located at the floor of the 3rd and 4th ventricle were recognized as reticulospinal neurons. Mauthner and Müller cells were identified according to morphological criteria. Eight pairs of Müller cells and one pair of Mauthner cells were labeled by NADPH histochemistry. None of these cells had, as yet, been described to display NOS activity in any vertebrate. The massive staining of these cells and the apparent lack of labeling, e.g., in teleost fishes, may be a histochemical correlate to already known differences of functions served by these cells in different species. In addition, our results suggest that the nitric oxide (NO) system has appeared early in vertebrate evolution.

Rhenium-188 for inhibition of human aortic smooth muscle cell proliferation.

PURPOSE: To evaluate dose-dependent growth-modulating effects of the beta-gamma emitter Rhenium-188 on cultured human aortic smooth muscle cells (haSMC). METHODS AND MATERIALS: HaSMC were plated in 25 cm(2) flasks. Two days after plating, cells were incubated with the Re-188 (beta E(max) 2.12 MeV, tissue range(max) < 10 mm, T(1/2) 17 h) for five days. The doses administered were 0.2 Gy, 1, 4, 6, 8, 16, and 32 Gy. After five days, the radionuclide was removed. Cell growth, cell cycle distribution, and clonogenic activity were analyzed for the following 25 days. RESULTS: The 0.2 and 1 Gy groups did not show relevant growth-inhibiting effects compared to the control groups. The 4 to 32 Gy groups presented dose-dependent growth inhibition, with a complete growth arrest of the 16 and 32 Gy groups. Clonogenic activity of the smooth muscle cell was strongly inhibited from doses > or =8 Gy. Flow cytometry showed a lasting dose-dependent G2/M phase block. CONCLUSION: Smooth muscle cell (SMC) growth can be controlled effectively with Re-188 for at least 25 days after radiation in vitro. As the first four weeks after arterial angioplasty are crucial concerning neointimal formation, Re-188 may be a valuable radionuclide to inhibit restenosis after arterial angioplasty.

Optimized fixation and storage conditions for FISH analysis of single-cell suspensions.

In our protocol to isolate and identify fetal cells in maternal peripheral blood, antibody (Ab)-stained cells are preserved with paraformaldehyde (PF) before batch flow cytometric sorting. However, PF fixation compromises the quality of subsequent interphase fluorescence in situ hybridization (FISH). We therefore examined the effect of PF concentrations and storage time in phosphate-buffered saline (PBS) on the quality of FISH signals. Cells were analyzed for changes in light scatter, morphology, and accessibility of target cell DNA. Fixation in 3% PF for 1 hr was ideal for both flow cytometry and subsequent FISH detection. However, beyond 10 days of storage, FISH quality deteriorated. (J Histochem Cytochem 46:971-973, 1998)

The basal forebrain cholinergic system in the raccoon.

The distribution of neurons displaying choline acetyltransferase (ChAT) immunoreactivity was examined in the raccoon basal forebrain using a rabbit antiserum and a monoclonal antibody. Alternating sections were used for Nissl staining. ChAT-positive neurons were arranged in a continuous mass extending from the medial septum to the caudal pole of the pallidum. Based upon spatial relations to fibre tracts, the clustering of neuronal groups, and cytological criteria, the basal forebrain magnocellular complex can be subdivided into several distinct regions. Although clear nuclear boundaries were often absent, the ChAT-positive neurons were divided into: the nucleus tractus diagonalis (comprising pars septi medialis, pars verticalis and pars horizontalis); nucleus praeopticus magnocellularis; substantia innominata; and the nucleus basalis of Meynert. Comparison with Nissl-stained sections indicated the presence of varying proportions of non-cholinergic neurons clustered or arranged loosely within these basal forebrain subdivisions. These data provide a structural basis for studies concerned with the topographical and physiological aspects of the raccoon basal forebrain cholinergic projections and its comparison with the basal forebrains of other species.

Long-term enhancement of evoked potentials in raccoon somatosensory cortex following co-activation of the nucleus basalis of Meynert complex and cutaneous receptors.

Long-term enhancement of the evoked potential was induced in the primary somatosensory cortex of anaesthetized raccoons after mechanical stimulation of the skin was paired with electrical stimulation of the nucleus basalis of Meynert (NBM). Sets of 4 pulses, 0.5 ms duration at 300 Hz were delivered at 2-s intervals to the basal forebrain 80 ms before the glabrous skin on the 4th digit of the contralateral forepaw was stimulated mechanically. The average waveform of 30 evoked potentials was separated into an initial positive, a negative and a second positive component. During pairing of the skin and NBM stimuli, the area under the initial positive component was smaller than before or after pairing. The negative and second positive waves were unchanged. One minute after pairing, the initial positive wave returned to control values and continued to increase until the end of the experiment 50 min later, at which time it was 300% above control. The negative and second positive waves increased after the pairing to between 130 and 200% and remained at that level for the duration of the experiment. The effective NBM site for stimulation was the area rich in cholinergic neurons corresponding to the NBM. In control animals, repeated stimulation of the skin or NBM alone, or their random, unpaired stimulation together, did not enhance the somatosensory evoked potential. The results suggest that the NBM input enhances the efficacy of cortical responses to cutaneous input and thus may play a role in cortical neuronal plasticity.

Development of the rabbit retina. V. The question of 'columnar units'.

A qualitative and quantitative description of the columnar units in the mammalian retina, and a discussion of their ontogeny and putative functions is given. Columnar arrangements of cells exist in the developing retina which can be observed by means of scanning electron microscopy. In the adult retina, each Müller cell ensheaths a columnar group of neuronal cells. Counting the number of cells in radial H/E stained sections at various developmental stages reveals a constant ratio of neuronal cells per Müller cell, independent of the developmental stage (after postnatal day 9), and independent of the retinal topography. Such groups of cells always consist of one Müller cell, 11 rod photoreceptor cells, about 2 bipolar cells, and 1 to 2 amacrine cells. Retinal ganglion cells, cone photoreceptor cells, and horizontal cells are more sparsely distributed in the retina than these units; since they are known to arise earlier in the ontogenesis than other cell types they are considered to exist independently of the columnar units. It is suggested that the units arise by migration of groups of preneurons along a common Müller (precursor) cell; these preneurons and the corresponding Müller cell may be clonally related. In the adult retina, such columns might constitute metabolic and functional units.

Development of the rabbit retina. I. Size of eye and retina, and postnatal cell proliferation.

Measures of rabbit eyes and retinal wholemounts were used to evaluate the development of retinal area and shape. The retina is shown to have a horizontal axis about a third longer than the vertical axis just before birth, and to adopt an almost symmetrical shape during postnatal development to adulthood. In general, retinal thickness is shown to decrease after birth, but differently in particular retinal regions: the reduction is marked in the periphery, and less pronounced in the visual streak. As an exception, the myelinated region--after it becomes really myelinated, from 9 days p.p.--even increases in thickness. In all regions of the retina, the absolute and relative thickness of the nuclear layers decreases, whereas the relative thickness of plexiform and fibrous layers increases. Proliferation of cells within the rabbit retina was studied during the first three postnatal weeks. 3H-thymidine incorporation was used to demonstrate DNA synthesis autoradiographically in histological sections as well as in enzymatically isolated retinal cells. A first proliferation phase occurs in the neuroblastic cell layer and ceases shortly after birth in the retinal center, but lasts for about one week in the retinal periphery. We found, however, a few 3H-thymidine-labeled cells as late as in the third postnatal week. These late-labeled cells were found within the nerve fiber layer and in the inner plexiform layer. The latter cells were shown to express antigens detected by antibodies directed to the intermediate-sized filament protein vimentin, which are known to label Müller cells and neuroepithelial stem cells. This was confirmed in our preparation of enzymatically isolated cells; all cells with autoradiographically labeled nuclei revealed a characteristic elongated morphology typical for Müller radial glia (and also for early neuroepithelial stem cells). 3H-thymidine-labeled cells in the nerve fiber layer were most probably astrocytic. In analogy to the brain, we conclude that the mammalian retina undergoes a series of proliferation phases: first an early phase producing both neurons and glial cells, and then a late phase producing glial cells, e.g., in the nerve fiber layer. Most probably, the late phase within the inner nuclear layer is glial as well, i.e., consists of dividing Müller cells; it cannot be excluded, however, that there may remain some mitotically active stem cells.

[Direct CT-arthrography versus direct MR-arthrography in chronic shoulder instability: comparison of modalities after the introduction of multidetector-CT technology]. / Direkte CT-Arthrographie versus direkte MR-Arthrographie bei chronischer Schulterinstabilität: Ein Methodenvergleich nach Einführung der Multidetektor-CT-Technik.

PURPOSE: To evaluate the diagnostic accuracy of direct multidetector CT arthrography (CTA) and direct MR arthrography (MRA) in patients suffering from chronic shoulder instability. MATERIALS AND METHODS: Twenty-nine patients suffering from chronic shoulder instability were included into a prospective study. In all cases, the indication for direct CTA and arthroscopy was set by the orthopedic surgeon. Prior to the imaging procedures, 10 to 20 ml of a special combination of contrast media (including saline, Isovist(R) and Magnevist(R) in a relation of 125 : 125 : 1) was injected into the joint under sterile conditions. First, CTA was performed with a multidetector CT, with images reconstructed in the axial, semi-coronal and semi-sagittal planes. Thereafter, MRA was performed. Axial images were obtained using a T1-weighted, fat-saturated spin echo sequence and semi-coronal images using a T1-weighted FLASH-3D GRE sequence. The results of CTA and MTA were compared with results obtained from arthroscopy or arthrotomy. RESULTS: MRA was superior to CTA in the detection of labral lesions. The sensitivity of MRA was 96 % and the specificity 96 %, compared to a sensitivity of 76 % (p < 0.05) and specificity of 92 % for CTA. Both methods showed the same effectiveness concerning the assessment of capsule distension (sensitivity for both techniques: 91 %). CONCLUSIONS: MRA seems to be superior to CTA in the diagnostic workup of chronic shoulder instability even when using a multidetector CT technique.

[Possibilities of the prophylaxis of arterial restenoses with dexamethasone -- an in-vitro study of human aortic smooth muscle cells]. / Möglichkeiten der Prophylaxe von arteriellen Restenosen mit Dexamethason -- eine In-vitro-Studie an humanen aortalen glatten Muskelzellen.

PURPOSE: To evaluate the effect of dexamethasone on the growth of cultured human aortic smooth muscle cells in an in-vitro model depending on the dose applied. MATERIALS AND METHODS: Commercially available human aortic smooth muscle cells (haSMC) were incubated with different doses of dexamethasone (10(-6), 10(-8), 10(-10) mol/l). For 20 days, the dose-depending effects of dexamethasone on cell growth were studied by analyzing cell proliferation, clonogenic activity as well as cell cycle distribution. In addition, the migratory ability of haSMC was evaluated using a two compartment in-vitro model. RESULTS: Cell growth was reduced in a dose dependent manner. An applied dose of 10(-6) M dexamethasone effectively inhibited cell growth for the follow-up period of 20 days. Cell cycle analysis revealed a G1-phase block which was dose dependent and significant for a dose of 10(-6) M. Also a reduction of haSMC clonogenic activity could be found in the colony formation assays. Finally, dexamethasone reduced the migratory ability of the treated cells significantly for doses of 10(-6) and 10(-8) M. CONCLUSION: Depending on the dose applied, incubation with dexamethasone results in a significant growth reduction of cultured haSMC, which may be due to a drug induced G1-phase block. Dexamethasone also reduces the clonogenic activity as well as the migratory ability of cultured haSMC.

[Dose-dependent effects of the combined beta/gamma emitter 186-Rhenium on the growth of human vessel wall cells]. / Dosisabhängige Auswirkungen des kombinierten beta/gamma Emitters Rhenium186 auf das Wachstum humaner Gefässwandzellen.

PURPOSE: The aim of this study was to evaluate the capability of human aortic smooth musc e cells (HaSMC) and endothelial cells (EC) to recover after incubation with the combined beta/gamma emitter 186rhenium. MATERIALS AND METHODS: Two days after plating, HaSMC and EC were incubated for five days with 186Re (total doses applied 4 Gy-32 Gy). Cell counts were performed for a period of 30 days (haSMC) and 22 days (EC). To detect possible growth recovery, colony formation assays were plated for both cell types on day 5, 10, and 20 (and lay 30 for haSMC). RESULTS: Both cell types presented a dose-dependent growth inhibition which was maximum at a dose of 32 Gy. Human endothelial cells presented with total growth recovery at 4 and 8 Gy, and a partial growth recovery at 16 Gy. Smooth muscle cells only presented partial growth recovery at 4 and 8 Gy. At 16 Gy and more no recovery was detected. CONCLUSION: HaSMC as well as EC growth can be modulated effectively with 186Re over a period of 30 days in vitro. Compared to smooth muscle cells human endothelial cellls seem to possess a higher potential to recover at doses of 8 to 16 Gy. 186Re may be a valuable radionuclide to prevent restenosis.

[Morpho-histochemical characterization of the connections between the dorsal part of the lateral geniculate nucleus (LGN) and the brain stem of albino rats) (author's transl)]. / Morphologisch-histochemische Charakterisierung der Verbindungen des Corpus geniculatum laterale, pars dorsalis (Cgl d), mit dem Hinrstamm bei Albinoratten.

After selective application of horseradish peroxidase (HRP) into the dorsal part of the lateral geniculate nucleus (LGN) of the albino rat, cells were labelled through the retrograde transport of the enzyme in the following nuclei of the brain stem: 1. Ncl. raphes linearis, 2. Nel, raphes centralis, 3. Ncl. raphes dorsalis, 4. Ncl. ventralis rostralis lemnisci lateralis, 5. Ncl. coeruleus. Using the glyoxylic acid method to demonstrate biogenic amines we could find serotonine in following areas of the brain stem: 1. region B 7 (Ncl. raphes dorsalis) positive cells; 2. region B 8 (Ncl. raphes centralis) mainly positive fibres; 3. region B 9 (dorsal of the Lemniscus medialis) a few positive cells; 4. mesencephalic reticular formation (Area cuneiformis) network-like fibre structures; 5. dorsal LGN, in the lateral part mildly fluorescing fibres, the remaining LGN showing diffuse fluorescence, 6. Substantia nigra, fluorescing cells. The monosynaptic connections between the mesencephalic raphe nuclei and the dorsal part of the LGN, demonstrated by means of the HRP-method, are characterized as being serotoninergic with the help of fluorescence histochemistry. The character of the transmitter of these connections is compared with results of biochemical and pharmacological investigations.