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Biomedical subjects

J Springer

Publications and source records attributed to J Springer.

At least 19 recordsLinked to original sources

Transcriptional up-regulation of histamine receptor-1 in epithelial, mucus and inflammatory cells in perennial allergic rhinitis.

BACKGROUND: Histamine receptors play an important role in the pathogenesis of nasal allergy. Activation of histamine receptor 1 (H1R) and 2 (H2R) can cause allergic symptoms which can be blocked effectively by antihistamines. H1R and H2R transcript levels have been found to be up-regulated in perennial - but not in seasonal - allergic rhinitis (AR). The present study aimed to explore H1R and H2R expression in complex tissues of the nasal mucosa of perennial allergic rhinitis (PAR). METHODS: Ten patients with PAR and 13 non-AR subjects were recruited for the study by medical history, physical examination and laboratory screening tests. In this study, we have analysed single cells dissected from the nasal mucosa biopsies by laser-assisted microdissection. H1R mRNA expression was analysed in different cell types such as epithelial, endothelial, mucus and inflammatory cells isolated from the nasal mucosa of PAR in comparison with non-AR subjects. RESULTS: H1R mRNA gene expression level was significantly increased in the nasal mucosa of PAR in comparison with non-AR (P<0.0001). H1R mRNA was significantly elevated in epithelial (P<0.001) and mucus cells (P<0.05) of PAR in comparison with non-AR whereas H1R gene expression levels in endothelial cells between both groups were not changed (P=0.23). Interestingly, inflammatory cells in the nasal mucosa of PAR patients were also strongly expressed H1R mRNA (P<0.001). CONCLUSION: The present study indicates that PAR alters the expression of H1R mRNA in epithelial, mucus and inflammatory cells of the nasal mucosa and but not in endothelial cells. Therefore, epithelial, mucus and inflammatory cells may play an important role in histamine-mediated allergic airway inflammation in PAR.

Adolescent↗

Immunohistochemical localization of cardio-active neuropeptides in the heart of a living fossil, Nautilus pompilius L. (Cephalopoda, Tetrabranchiata).

Neuropeptides play an important role in modulating the effects of neurotransmitters such as acetylcholine and noradrenaline in the heart and the vascular system of vertebrates and invertebrates. Various neuropeptides, including substance P (SP), vasoactive intestinal polypeptide (VIP) and FMRFamide, have been localized in the brain in cephalopods and the neurosecretory system of the vena cava. Previous studies involving cephalopods have mainly focussed on the modern, coleoid cephalopods, whereas little attention was paid to the living fossil Nautilus. In this study, the distributions of the peptides related to tachykinins (TKs) and the high affinity receptor for the best characterized TK substance P (tachykinin NK-1), VIP, as well as FMRFamide were investigated in the heart of Nautilus pompilius L. by immunohistochemistry. TK-like immunoreactivity (TK-LI) was seen associated to a sub-population of hemocytes, VIP-LI glial cells in larger nerves entering the heart, whereas FMRFamide immunoreactivity was distributed throughout the entire heart, including the semilunar atrioventricular valves. The pattern of FMRFamide immunoreactivity matched that of Bodian silver staining for nervous tissue. The NK-1-LI receptor was located on endothelial cells, which were also positive for endothelial nitric oxide synthase-LI (eNOS). The results indicate that neuropeptides may be involved in the regulation of the Nautilus heart via different mechanisms, (1) by direct interaction with myocardial receptors (FMRFamide), (2) by interacting with the nervus cardiacus (VIP-related peptides) and (3) indirectly by stimulating eNOS in the endothelium throughout the heart (TK-related peptides).

Animals↗

Nerve growth factor-induced substance P in capsaicin-insensitive vagal neurons innervating the lower mouse airway.

BACKGROUND: Nerve growth factor (NGF) is elevated in allergic diseases such as bronchial asthma and can lead to an induction of substance P (SP) and related neuropeptides in guinea-pigs large-diameter, neurofilament-positive airway neurons. OBJECTIVE: In the present study, the effect of NGF on tyrosine kinase receptor trkA and the capsaicin receptor TRPV1 expression in airway-specific vagal sensory neurons located in the jugular-nodose ganglia complex (JNC) of mice was investigated. METHODS: Using retrograde neuronal tracing in combination with double-labelling immunohistochemistry, SP, trkA- and TRPV1-receptor expression was examined in airway-specific sensory neurons of BALB/c mice before and after NGF treatment. RESULTS: NGF injected into the lower airway was able to induce SP (13.0+/-2.03% vs. 5.9+/-0.33%) and trkA expression (78+/-2.66% vs. 60+/-2.11%) in larger diameter (>25 microm), capsaicin-insensitive and trkA-positive vagal sensory neurons that were retrograde-labelled with Fast Blue dye from the main stem bronchi. CONCLUSION: Based on the extent of SP and trkA co-expression in airway-specific neurons by NGF treatment, the present study suggests that, following a peripheral activation of trkA receptor on SP afferent by NGF which is elevated in allergic inflammation, there may be trkA-mediated SP induction to mediate neurogenic airway inflammation.

Animals↗

Effective mass of two-dimensional 3He.

We use structural information from diffusion Monte Carlo calculations for two-dimensional 3He to calculate the effective mass. Static effective interactions are constructed from the density and spin-structure functions using sum rules. We find that both spin and density fluctuations contribute about equally to the effective mass. Our results show, in agreement with recent experiments, a flattening of the single-particle self-energy with increasing density, which eventually leads to a divergent effective mass.

Journal Article↗

[Internal rotational strength of the thigh after ACL reconstruction. Prospective comparison of harvesting of the semitendinosus and gracilis tendons and the patella tendon].

AIM: The aim of the present study was the prospective, randomized evaluation of the internal rotation strength in patients, undergoing ACL reconstruction using either semitendinosus and gracilis tendon autografts (ST/G) or the central third of the patella tendon (BPT). METHOD: 40 patients (20 male, 20 female, average age 32 years [16 - 49]) underwent reconstruction of the ACL. In 20 patients ST/G was used, in 20 matched patients the central third of the patella tendon (BPT) was used. Rehabilitation was identical for both groups. The isometric strength of internal tibial rotation in neutral position was measured preoperatively, 3, 6 and 12 months postoperatively. RESULTS: After 3 months no difference to the preoperative data was found. After six months the internal tibial torque was significantly higher (p < 0.05) than preoperatively. No additional increase was found after 12 months. There was no statistically significant difference between both groups at any time. CONCLUSION: The internal tibial torque, which is predominantly a function of the pes anserinus, recovers shortly after operation and is, due to postoperative rehabilitation, significantly elevated after 6 months. The randomized comparison with the harvesting of the central third of the patellar tendon showed no significant difference.

Adolescent↗

Substance P-induced pulmonary vascular remodelling in precision cut lung slices.

Substance P (SP) levels have been reported to be elevated in animal models of pulmonary hypertension (PH) and chronic administration of SP has been shown to induce PH. In the present study, the role of reactive oxygen species (ROS) as mediators of SP-induced vascular remodelling and PH was analysed. Vascular remodelling was induced in precision cut lung slices by treatment with [Sar9,Met11(O2)]-SP and by hypoxia. Functional analyses were used to study the chronic [Sar9,Met11(O2)]-SP-mediated effects on proliferation and generation of ROS, which is involved in the pathogenesis of PH. The nonpeptide NK-1 receptor antagonist CP 96345 was used to block [Sar9,Met11(O2)]-SP effects. ROS generation and proliferation was assayed by the dichloroflourescein-diacetate method and incorporation of 5-Bromo-2'-Deoxyuridine, respectively. ROS generation was induced by either 1% oxygen (5.8-fold) or [Sar9,Met11(O2)]-SP (8-fold) in normoxia in the alveolar region. [Sar9,Met11(O2)]-SP did not further elevate ROS levels in hypoxia, suggesting an oxygen-dependent mechanism. High ROS levels stabilised hypoxia inducible factor-1alpha and induced proliferation in small vessels (4.3-fold in hypoxia and [Sar9,Met11(O2)]-SP). Both ROS generation and proliferation were blocked in the presence of CP 96345, nitroblue-tetrazolium, N-acetylcysteine- and diphenylene-iodonium. The results presented in this study indicate a role of SP in proliferative events associated with vascular remodelling in pulmonary hypertension.

Animals↗

Abundant expression of c-Jun in guinea pig sympathetic ganglia under basal conditions and allergen challenge.

Airway hyperresponsiveness, a keystone of allergic asthma, is mediated by the extrinsic airway innervation. As pathophysiological stimuli can induce the expression JUN proteins, which belong to the immediate early gene (IEG) family of transcription factors, the expression of c-Jun was examined under basal conditions and allergen challenge in guinea pig paravertebral and prevertebral sympathetic ganglia by quantitative double-labeling immunohistochemistry. C-Jun immunoreactivity was seen in 78.4 +/- 3.5% under normal and 82.6 +/- 4.6% under allergen-challenged conditions of protein-gene product (PGP) 9.5-positive sympathetic neurons of guinea pig superior cervical ganglia and 73.1 +/- 2.8% (normal) and 76.1 +/- 3.5% (allergen) of stellate ganglion neurons. In the coeliac-superior mesenteric ganglion, 59.5 +/- 5.0% (normal) and 57.5 +/- 4.4% (allergen) of the PGP 9.5-positive sympathetic neurons were labeled for c-Jun. The high basal levels of c-Jun expression indicate that the presence of c-Jun is not exclusively related to noxious stimulation such as allergic airway inflammation in the guinea pig.

Animals↗

Vasoactive intestinal polypeptide as mediator of asthma.

Vasoactive intestinal polypeptide (VIP) is one of the most abundant, biologically active peptides found in the human lung. VIP is a likely neurotransmitter or neuromodulator of the inhibitory non-adrenergic non-cholinergic airway nervous system and influences many aspects of pulmonary biology. In human airways VIP-immunoreactive nerve fibres are present in the tracheobronchial airway smooth muscle layer, the walls of pulmonary and bronchial vessels and around submucosal glands. Next to its prominent bronchodilatory effects, VIP potently relaxes pulmonary vessels. The precise role of VIP in the pathogenesis of asthma is still uncertain. Although a therapy using the strong bronchodilatory effects of VIP would offer potential benefits, the rapid inactivation of the peptide by airway peptidases has prevented effective VIP-based drugs so far and non-peptide VIP-agonists did not reach clinical use.

Airway Resistance↗

Localization of the peptide transporter PEPT2 in the lung: implications for pulmonary oligopeptide uptake.

Pulmonary delivery of peptidomimetic antibiotics is frequently used for local drug therapy in pulmonary infections. Identification of transport pathways into airway epithelia can lead to new strategies of therapy. Here we describe the distribution of the beta-lactam-transporting high-affinity proton-coupled peptide transporter PEPT2 in mammalian lungs. Using reverse transcriptase-polymerase chain reaction and Northern blot analysis, PEPT2-mRNA was detected in lung extracts. The expression of PEPT2-mRNA and protein was localized to alveolar type II pneumocytes, bronchial epithelium, and endothelium of small arteries of rat lung by nonisotopic in situ hybridization and immunohistochemistry. In addition, transport studies using murine whole-organ preparations revealed transporter-mediated uptake of a fluorophore-conjugated dipeptide derivative into bronchial epithelial cells and type II pneumocytes. This transport was competitively inhibited by cephalosporins and dipeptides that are reported as PEPT2-carried substrates. Cell specificity of the PEPT2-mediated uptake pattern was confirmed by double labeling with Lycopersicon esculentum lectin. Together these data suggest that PEPT2 is the molecular basis for the transport of peptides and peptidomimetics in pulmonary epithelial cells. In conclusion PEPT2 may be an interesting target for pulmonary delivery of peptides and peptidomimetics.

Animals↗

Expression of immediate early genes in sensory ganglia.

C-Jun and c-Fos belong to the family of immediate early genes. Apart from their role as transcription factors, a basal expression was shown for them in central nervous system tissues. The expression of c-Jun and c-Fos in sensory ganglia of guinea pig, rat and murine sensory ganglia was examined under normal, unstimulated conditions by quantitative double-immunohistochemistry. 4.6 +/- 2.8% of neuron-specific protein gene-product 9.5 -positive cells in nodose ganglia, 51.6 +/- 2.1% in jugular ganglia, 46.4 +/- 3.0% in trigeminal ganglia and 42.5 +/- 1.3% of cervical dorsal root ganglia neurons were positive for c-Jun in the guinea pig (less than 1% for c-Fos). In rat and mouse, less than 1% of the sensory neurons exhibited c-Jun and c-Fos-immunoreactivity. The high basal expression of c-Jun in guinea pig sensory neurons suggests that in this species the presence of c-Jun does not only depend on specific stimulation and is not exclusively associated with neuronal plasticity of gene expression and functional changes.

Animals↗

Immunohistochemical detection of calcitonin gene-related peptide receptor (CGRPR)-1 in the endothelium of human coronary artery and bronchial blood vessels.

The potent vasodilatory peptide, calcitonin gene-related peptide (CGRP) is present in the innervation of vascular tissue. The actions of CGRP occur via a receptor, CGRP receptor(R)-1, which is also a target for the cardioprotective peptide adrenomedullin. The human version of the pharmacologically-defined CGRPR-1 has been cloned but its distribution and cellular location is unknown. A rabbit antibody was generated to a synthetic peptide that corresponds to the C-terminus of human CGRPR-1 Immunochemical analysis of the human cell-line, SK-N-MC, which exhibits functional expression of the CGRPR-1 confirmed the antibody's specificity. The antiserum revealed specific staining in the endothelium of human coronary arteries. The vascular smooth muscle and ventricular myocardium were not immunoreactive. In bronchial blood vessels CGRPR-1-immunoreactivity was detected in the endothelium of the venules and not in the arterioles, which is particularly relevant for elucidating the putative role of CGRP in inflammation in this tissue.

Antibody Specificity↗

Effect of introns and AT-rich sequences on expression of the bacterial hygromycin B resistance gene in the basidiomycete Schizophyllum commune.

Previously, it was shown that introns are required for efficient mRNA accumulation in Schizophyllum commune and that the presence of AT-rich sequences in the coding region of genes can result in truncation of transcripts in this homobasidiomycete. Here we show that intron-dependent mRNA accumulation and truncation of transcripts are two independent events that both affect expression of the bacterial hygromycin B resistance gene in S. commune.

AT Rich Sequence↗

NK1 receptor stimulation causes contraction and inositol phosphate increase in medium-size human isolated bronchi.

Although contraction of human isolated bronchi is mediated mainly by tachykinin NK2 receptors, NK1 receptors, via prostanoid release, contract small-size (approximately 1 mm in diameter) bronchi. Here, we have investigated the presence and biological responses of NK1 receptors in medium-size (2-5 mm in diameter) human isolated bronchi. Specific staining was seen in bronchial sections with an antibody directed against the human NK1 receptor. The selective NK1 receptor agonist, [Sar(9), Met(O2)(11)]SP, contracted about 60% of human isolated bronchial rings. This effect was reduced by two different NK1 receptor antagonists, CP-99,994 and SR 140333. Contraction induced by [Sar(9), Met(O2)(11)]SP was independent of acetylcholine and histamine release and epithelium removal, and was not affected by nitric oxide synthase and cyclooxygenase (COX) inhibition. [Sar(9), Met(O2)(11)]SP increased inositol phosphate (IP) levels, and SR 140333 blocked this increase, in segments of medium- and small-size (approximately 1 mm in diameter) human bronchi. COX inhibition blocked the IP increase induced by [Sar(9), Met(O2)(11)]SP in small-size, but not in medium-size, bronchi. NK1 receptors mediated bronchoconstriction in a large proportion of medium-size human bronchi. Unlike small-size bronchi this effect is independent of prostanoid release, and the results are suggestive of a direct activation of smooth muscle receptors and IP release.

Analysis of Variance↗

Species of the toxic Pfiesteria complex, and the importance of functional type in data interpretation.

We describe the two species of the toxic Pfiesteria complex to date (Pfiesteria piscicida and Pfiesteria shumwayae), their complex life cycles, and the characteristics required for inclusion within this complex. These species resemble P. piscicida Steidinger & Burkholder and also have a) strong attraction to fresh fish tissues and excreta, b) toxic activity stimulated by live fish, and c) production of toxin that can cause fish death and disease. Amoeboid stages were verified in 1992-1997 by our laboratory (various stages from toxic cultures) and that of K. Steidinger and co-workers (filose amoebae in nontoxic cultures), and in 2000 by H. Marshall and co-workers (various stages from toxic cultures), from clonal Pfiesteria spp. cultures, using species-specific polymerase chain reaction-based molecular probes with cross-confirmation by an independent specialist. Data were provided from tests of the hypothesis that Pfiesteriastrains differ in response to fresh fish mucus and excreta, algal prey, and inorganic nutrient (N, P) enrichment, depending on functional type or toxicity status. There are three functional types: TOX-A, in actively toxic, fish-killing mode; TOX-B, temporarily nontoxic, without access to live fish for days to weeks, but capable of toxic activity if fish are added; and NON-IND, noninducible with negligible toxicity in the presence of live fish. NON-IND Pfiesteria attained highest zoospore production on algal prey without or without inorganic nitrogen or inorganic phosphorus enrichment. TOX-B Pfiesteria was intermediate and TOX-A was lowest in zoospore production on algal prey with or without nutrients. TOX-A Pfiesteria spp. showed strong behavioral attraction to fresh fish mucus and excreta in short-term trials, with intermediate attraction of TOX-B zoospores and relatively low attraction of NON-IND cultures when normalized for cell density. The data for these clones indicated a potentially common predatory behavioral response, although differing in intensity distinct from a toxicity effect, in attack of fish prey. The data also demonstrated that functional types of Pfiesteria spp. show distinct differences in response to fish, algal prey, and inorganic nutrient enrichment. Collectively, the experiments indicate that NON-IND strains should not be used in research to gain insights about environmental controls on toxic strains of Pfiesteria spp.

Animals↗

Cloning and characterization of the Yarrowia lipolytica squalene synthase (SQS1) gene and functional complementation of the Saccharomyces cerevisiae erg9 mutation.

The squalene synthase (SQS) gene encodes a key regulatory enzyme, farnesyl-diphosphate farnesyltransferase (EC 2.5.1.21), in sterol biosynthesis. The SQS1 gene was isolated from a subgenomic library of the industrially important yeast Yarrowia lipolytica, using PCR-generated probes. Probes were based on conserved regions of homologues from different organisms. The complete nucleotide sequence of the coding region and the corresponding amino acid sequence were determined. The sequences showed extensive homologies with squalene synthase genes and enzymes from a number of other organisms and extreme amino acid conservation within the binding and catalytic domains. Direct cloning of a 4.3 kb genomic Y. lipolytica fragment, also comprising its own promoter and terminator sequences, into autonomously replicating plasmid YEp352 and subsequent transformation of a Saccharomyces cerevisiae mutant strain with relevant erg9: ura3-1 markers, resulted in functional complementation of these deficiencies, although Northern blot analyses did not reveal a unique full-length messenger. The availability of the Y. lipolytica SQS1 gene (GenBank Accession No. AF092497) offers prospects for metabolic engineering of the isoprenoid and sterol biosynthetic pathways.

Amino Acid Sequence↗

Characterization of Several Polymer Surfaces by Streaming Potential and Wetting Measurements: Some Reflections on Acid-Base Interactions.

Several thermoplastic (technical, engineering, and high-performance) polymers were characterized using contact angle and electrokinetic measurements. From the measured contact angles of various test liquids on polymers, we calculated the solid surface tensions using the different approaches to determine them and compared the results. Zeta (zeta)-potential measurements gave information about the swelling behavior of the polymers in water, the surface chemistry, and the interactions with dissolved potassium and chloride ions. All investigated polymers displayed an acidic surface character. Comparing the results obtained from the zeta-potential measurements with the acid-parameter of the surface tension gamma(+) calculated from the measured "static" contact angles using the van Oss, Good, and Chaudhury approach revealed the same tendency. The correctness of the acid-base approach regarding the "overall" chemical surface character could be shown. However, it seems that the basic parameter gamma(-) obtained from the acid-base is greatly overestimated. Copyright 1999 Academic Press.

Journal Article↗