Biomedical subjects

S Zimmermann

Publications and source records attributed to S Zimmermann.

At least 19 recordsLinked to original sources

PubMed2000

CpG-DNA-mediated transient lymphadenopathy is associated with a state of Th1 predisposition to antigen-driven responses.

Infections can influence concurrent and subsequent Th1 vs Th2 immune responses to Ags. Through pattern recognition of foreign unmethylated CpG dinucleotides, the vertebrate innate immune system can sense infectious danger and typically replies with a Th1-polarized adaptive immune response. We examined whether CpG-DNA exposure would influence subsequent responses to infection and soluble Ags. CpG-DNA injection led to local lymphadenopathy characterized by maintenance of cellular composition with some biasing toward elevated dendritic cell composition. Sustained local production of IL-12 and IFN-gamma from dendritic cells and T cells was shown. Prior injection by up to 2 wk with CpG-DNA protected BALB/c mice from Th2 driven lethal leishmaniasis. CpG-DNA injection by up to 5 wk before soluble Ag challenge resulted in the generation of Ag-specific CTL, Th1 recall responses to Ag, and Th1-polarized Ag-specific Abs. Thus, CpG-DNA instigated a local predisposition for intense CTL responses and Th1-polarized immune responses to subsequent infections or Ag challenge. The induction by the innate immune system of a locally contained hypersensitivity could represent a capacitating immune reaction yielding rapid conditioned responses to secondary infections.

Adoptive Transfer↗

PubMed2000

Disruption of feeding behavior in CRH receptor 1-deficient mice is dependent on glucocorticoids.

Corticotropin-releasing hormone (CRH) has been found to markedly suppress food intake and reduce body weight. However, it still remains to be clarified whether those effects are mediated via either the CRH receptor 1 (CRHR1) or the CRH receptor 2 (CRHR2), or both receptor subtypes. Therefore, we investigated whether CRHR1-deficient mice (CRHR1-KO) show abnormalities in body weight and feeding behavior. CRHR1-KO and wildtype mice showed no difference in the total amount of food intake. However, there was a significant disruption in the circadian distribution of food intake: CRHR1 mutants consumed significantly more food during the light period (p<0.01). The normal diurnal pattern could be completely restored by oral administration of corticosterone 21-sulfate (5 mg/l added to the water-based liquid diet). We therefore conclude that in CRHR1-KO mice, the disruption of feeding behavior might be causally related to glucocorticoid deficiency, but that the CRHR1 is not likely to play a critical role in the basal regulation of ingestive behavior.

Administration, Oral↗

PubMed2000

Critical examination of trace element enrichments and depletions in soils: As, Cr, Cu, Ni, Pb, and Zn in Swiss forest soils.

The aim of this study was to obtain an overview of trace element concentrations in Swiss forest soils and to critically assess the measured values with respect to anthropogenic input vs. lithogenic background. Twenty-three sites were selected which represent a broad range of natural forest sites, bedrock material and soil types of Switzerland. At each site, samples were collected from all genetic soil horizons down to a C or B/C horizon. Total concentrations of As, Cr, Cu, Ni, Pb, and Zn in all samples were determined by X-ray fluorescence spectrometry. There were distinct differences in the geological background values estimated from the concentrations measured in the samples from the lowest soil horizon. Background concentrations for Cr and Ni were lowest in granite and gneiss, whereas Pb and Zn were highest in limestone and marl. Enrichment or depletion of the trace elements was assessed using Zr as reference element. Within the same profile, the six trace elements showed completely different enrichment/depletion patterns with depth. The various natural processes and anthropogenic inputs that can lead to these patterns are critically discussed. Based on this critical assessment, pollution of the investigated forest soils was found to be most severe for Pb and Zn and to a somewhat lesser extent for As and Cu, whereas anthropogenic input of Cr and Ni seems to be less important. The data suggest that a critical evaluation of enrichment factors is a better tool to assess soil pollution with trace elements than the use of maximum allowable concentrations (MAC) for topsoil samples. The enrichment factors calculated as described here consider the effects of geological variation on metal abundances whereas the MAC does not. In order to obtain an estimate of soil solution concentrations, water extracts of the samples collected from a subset of 10 soil profiles were analyzed for the same trace elements. Solubility of all elements generally decreased with soil depth. An exception was Cr, Cu, and Ni solubility in the humus layer, which was lower than in the underlying mineral horizon. For all elements, solubility was higher for the collective of soil samples depleted in this element when compared to the samples, in which the element was enriched.

Arsenic↗

PubMed2000

Functional characterisation of LKT1, a K+ uptake channel from tomato root hairs, and comparison with the closely related potato inwardly rectifying K+ channel SKT1 after expression in Xenopus oocytes.

A cDNA encoding a novel inwardly rectifying potassium (K+(in)) channel, LKT1, was cloned from a root-hair-specific cDNA library of tomato (Lycopersicon esculentum Mill.). The LKT1 mRNA was shown to be most strongly expressed in root hairs by Northern blot analysis. The LKT1 channel is a member of the AKT family of K+(in) channels previously identified in Arabidopsis thaliana (L.) Heynh. and potato (Solanum tuberosum L.). Moreover, LKT1 is closely related (97% identical amino acids) to potato SKT1. An electrophysiological comparison of the two channels should therefore assist the identification of possible molecular bases for functional differences. For, this comparison, both channels were functionally expressed and electrophysiologically characterised within the same expression system, i.e. Xenopus laevis oocytes. Voltage-clamp measurements identified LKT1 as a K(+)-selective inward rectifier which activates with slow kinetics upon hyperpolarising voltage pulses to potentials more negative than -50 mV. The activation potential of LKT1 is shifted towards positive potentials with respect to SKT1 which might be due to single amino acid exchanges in the rim of the channel's pore region or in the S4 domain. Like SKT1, LKT1 reversibly activated upon shifting the external pH from 6.6 to 5.5, which indicates a physiological role for pH-dependent regulation of AKT-type K+(in) channels. The pharmacological inhibitor Cs+, applied externally, inhibited K+(in) currents mediated by LKT1 and SKT1 half-maximally with a concentration (IC50) of 21 microM and 17 microM, respectively. In conclusion, LKT1 may serve as a low-affinity influx pathway for K+ into root hair cells. Comparison of homologous K+(in) rectifiers from different plant species expressed in the same heterologous system allows conclusions to be drawn in respect to structure-function relationships.

Amino Acid Sequence↗

PubMed2000

CpG DNA as a Th1 trigger.

Over the last few years, it has been recognized that along with structural components and products of bacteria, bacterial DNA is also capable of signaling infectious danger to cells of the innate immune system. Particular DNA sequences (CpG motifs), which are abundant in prokaryotic (bacterial) but not in mammalian DNA, cause the activation and stimulation of immune cells. Research has been catalyzed by the finding that certain synthetic oligodeoxynucleotides mimic the action of bacterial DNA. Immunostimulation induced by bacterial DNA or synthetic oligonucleotides not only contributes to our knowledge of the pathogen-host interrelationship during infection, but can also be used therapeutically to condition or modify ongoing immune responses of the adaptive immune system. Accordingly, CpG motifs have been used as vaccine adjuvants as well as instructing agents to selectively induce Th1-dominated immune responses. Hence, CpG motifs might be used in the future as adjuvants and/or immunomodulatory agents to treat or prevent undesired Th2-dominated immune responses, such as allergy.

Adjuvants, Immunologic↗

PubMed2000

Muscarinic control of histamine release from airways. Inhibitory M1-receptors in human bronchi but absence in rat trachea.

Isolated human bronchi and rat tracheae were incubated in organ baths to measure histamine release. The calcium ionophore A23187, 3 micromol/L in rat trachea and 10 micromol/L in human bronchi, stimulated histamine release by 145 +/- 50% (n = 6) and 270 +/- 48% (n = 7) above the prestimulation level, respectively. Acetylcholine (100 pmol/L; human bronchi) or oxotremorine (1, 100, 10,000 nmol/L; rat trachea) did not affect the spontaneous histamine release. In rat tracheae neither acetylcholine nor oxotremorine inhibited A23187-evoked histamine release, whereas 100 pmol/L acetylcholine significantly suppressed the evoked histamine release in human bronchi by 86%. For receptor characterization the following subtype-specific muscarinic receptor antagonists were applied: pirenzepine (M1 subtype), para-fluorohexahydrosiladifendiol (pFHHSiD; similar affinities at human cloned M1-, M3-, and M4-receptors), AF-DX 116 (M2 subtype), and clozapine (antagonist at cloned M1-, M2-, M3-receptors; agonist at cloned M4-receptors). Pirenzepine, pFHHSiD, AF-DX 116, and clozapine (100 nmol/L each) antagonized the inhibitory effect of 100 pmol/L acetylcholine by 83 +/- 20% (n = 6), 83 +/- 9% (n = 8), 50 +/- 14% (n = 6), and 35 +/- 7% (6). In conclusion, a species difference exists in the cholinergic control of histamine release between human and rat airways. In human airways muscarinic receptors most likely of the M1 subtype are involved in the inhibitory control of mast cell function, whereas such an inhibitory pathway does not exist in the rat trachea.

Acetylcholine↗

PubMed1999

Differentiation stage-specific inhibition of the Raf-MEK-ERK pathway by Akt.

Extracellular signals often result in simultaneous activation of both the Raf-MEK-ERK and PI3K-Akt pathways (where ERK is extracellular-regulated kinase, MEK is mitogen-activated protein kinase or ERK kinase, and PI3K is phosphatidylinositol 3-kinase). However, these two signaling pathways were shown to exert opposing effects on muscle cell hypertrophy. Furthermore, the PI3K-Akt pathway was shown to inhibit the Raf-MEK-ERK pathway; this cross-regulation depended on the differentiation state of the cell: Akt activation inhibited the Raf-MEK-ERK pathway in differentiated myotubes, but not in their myoblast precursors. The stage-specific inhibitory action of Akt correlated with its stage-specific ability to form a complex with Raf, suggesting the existence of differentially expressed mediators of an inhibitory Akt-Raf complex.

Animals↗

PubMed1999

Phosphorylation and regulation of Raf by Akt (protein kinase B).

Activation of the protein kinase Raf can lead to opposing cellular responses such as proliferation, growth arrest, apoptosis, or differentiation. Akt (protein kinase B), a member of a different signaling pathway that also regulates these responses, interacted with Raf and phosphorylated this protein at a highly conserved serine residue in its regulatory domain in vivo. This phosphorylation of Raf by Akt inhibited activation of the Raf-MEK-ERK signaling pathway and shifted the cellular response in a human breast cancer cell line from cell cycle arrest to proliferation. These observations provide a molecular basis for cross talk between two signaling pathways at the level of Raf and Akt.

Cell Division↗

PubMed1999

UV-responsive genes of arabidopsis revealed by similarity to the Gcn4-mediated UV response in yeast.

A UV response that involves the Ras proteins and AP-1 transcription factors has recently been described in mammals and yeast. To test whether an equivalent response exists in plants, we monitored the expression of Arabidopsis histidinol dehydrogenase gene (HDH), a homologue of the yeast HIS4 gene, which is strongly induced by UV light and is a target of the transcriptional activator Gcn4. We show that HDH mRNA levels increase specifically in response to UV-B light. Only small increases were detected upon exposure to other wavelengths. To isolate plant genes involved in this UV response, a gcn4 mutant was transfected with an Arabidopsis thaliana cDNA library. A new type of nucleotide diphosphate kinase (NDPK Ia) with a significant homology to the human tumor suppressor protein Nm23 rescued the gcn4 phenotype. NDPK Ia specifically binds to the HIS4 promoter in vitro and induces HIS4 transcription in yeast. In Arabidopsis, the NDPK Ia protein is located in the nucleus and cytosol. Expression studies in seedlings revealed that the level of NDPK Ia mRNA, like that of HDH, increases in response to UV-B light. It appears that NDPK Ia and HDH are components of a novel UV-responsive pathway in A. thaliana.

Alcohol Oxidoreductases↗

PubMed1999

A Semiconductor-Based Photonic Memory Cell.

Photonic signals were efficiently stored in a semiconductor-based memory cell. The incident photons were converted to electron-hole pairs that were locally stored in a quantum well that was laterally modulated by a field-effect tunable electrostatic superlattice. At large superlattice potential amplitudes, these pairs were stored for a time that was at least five orders of magnitude longer than their natural lifetime. At an arbitrarily chosen time, they were released in a short and intense flash of incoherent light, which was triggered by flattening the superlattice amplitude.

Journal Article↗

PubMed1999

Experimental studies on lead accumulation in the eel-specific endoparasites Anguillicola crassus (Nematoda) and Paratenuisentis ambiguus (Acanthocephala) as compared with their host, Anguilla anguilla.

The effect of salinity and the mode of application (oral versus aqueous) on the lead accumulation in different tissues of the fish host eel (Anguilla anguilla) and its parasites Anguillicola crassus (Nematoda) and Paratenuisentis ambiguus (Acanthocephala) was investigated. Waterborne as well as dietary lead exposure caused an increase in the metal levels of different eel tissues and the parasites. The mode of lead application had a significant influence on the distribution of lead in the fish tissues. No significant difference on the lead concentration due to water salinity was found for the fish tissues. Among the analyzed tissues and helminths, the intestinal acanthocephalan P. ambiguus contained the significantly highest amounts of lead, which were affected by neither the mode of application nor the water salinity. In contrast, the lead level of the nematode A. crassus dwelling in the swim bladder of eels was even below the levels detected for host liver, intestine, and bile. Thus, depending on the mode of lead application, the resulting metal concentrations were approximately 20 to 2,000 times higher in P. ambiguus than in A. crassus. These differences may be due to the different microhabitats and nutrient uptake mechanisms of both parasite species. This study presents important new facts for the use of intestinal fish parasites as biological indicators for water quality, not only in freshwater, but also in marine and estuarine ecosystems. The combination of the results obtained from the host and the parasites could reveal a more detailed tool to ascertain the source of an environmental contamination than a study based on a single species.http://link.springer-ny. com/link/service/journals/00244/bibs/37n2p190.html

Acanthocephala↗

PubMed1999

Plant ion channels: from molecular structures to physiological functions.

Progress in identification of plant ion channels and development of electrophysiological analyses in heterologous expression systems and in planta, in combination with reverse genetic approaches, are providing the possibility of associating molecular entities with physiological functions. Recently, the first attempts to determine in vivo functions using knockout mutants demonstrated the roles of root ion channels. The search for proteins interacting with such channels leads to an even more complex view of the concerted action in protein networks.

Ion Channels↗

PubMed1999

Apoplastic and cytosolic expression of full-size antibodies and antibody fragments in Nicotiana tabacum.

We compared the expression of a functional recombinant TMV-specific full-size antibody (rAb29) in both the apoplast and cytosol of tobacco plants and a single chain antibody fragment (scFv29), derived from rAb29, was expressed in the cytosol. Cloned heavy and light chain cDNAs of full-size rAb29, which binds to TMV coat protein monomers, were integrated into the plant expression vector pSS. The full-size rAb29 was expressed in the cytosol and targeted to the apoplast by including the original murine antibody leader sequences. Levels of functional full-size rAb29 expression were high in the apoplast (up to 8.5 micrograms per gram leaf tissue), whereas cytosolic expression was low or at the ELISA detection limit. Sequences of the variable domains of rAb29 light and heavy chain were used to generate the single chain antibody of scFv29, which was expressed in the periplasmic space of E. coli and showed the same binding specificity as full-size rAb29. In addition, scFv29 was functionally expressed in the cytosol of tobacco plants and plant derived scFv29 maintained same binding specificity to TMV-coat protein monomers as rAb29.

Antibodies, Viral↗

PubMed1999

Expression and characterization of bispecific single-chain Fv fragments produced in transgenic plants.

We describe the expression of the bispecific antibody biscFv2429 in transgenic suspension culture cells and tobacco plants. biscFv2429 consists of two single-chain antibodies, scFv24 and scFv29, connected by the Trichoderma reesi cellobiohydrolase I linker. biscFv2429 binds two epitopes of tobacco mosaic virus (TMV): the scFv24 domain recognizes neotopes of intact virions, and the scFv29 domain recognizes a cryptotope of the TMV coat protein monomer. biscFv2429 was functionally expressed either in the cytosol (biscFv2429-cyt) or targeted to the apoplast using a murine leader peptide sequence (biscFv2429-apoplast). A third construct contained the C-terminal KDEL sequence for retention in the ER (biscFv2429-KDEL). Levels of cytoplasmic biscFv2429 expression levels were low. The highest levels of antibody expression were for apoplast-targeted biscFv2429-apoplast and ER-retained biscFv2429-KDEL that reached a maximum expression level of 1.65% total soluble protein in transgenic plants. Plant-expressed biscFv2429 retained both epitope specificities, and bispecificity and bivalency were confirmed by ELISA and surface plasmon resonance analysis. This study establishes plant cells as an expression system for bispecific single-chain antibodies for use in medical and biological applications.

Antibodies, Bispecific↗

PubMed1999

Targeted disruption of the Insl3 gene causes bilateral cryptorchidism.

The sexual dimorphic position of the gonads in mammals is dependent on differential development of two ligaments, the cranial suspensory ligament (CSL) and the gubernaculum. During male embryogenesis, outgrowth of the gubernaculum and regression of the CSL result in transabdominal descent of the testes, whereas in the female, development of the CSL in conjunction with failure of the gubernaculum development holds the ovaries in a position lateral to the kidneys. Several lines of evidence suggest that regression of the CSL and induction of gubernaculum development are mediated by testosterone and a yet unidentified testicular factor, respectively. The Insl3 gene (originally designated Ley I-L), a member of the insulin-like superfamily, is specifically expressed in Leydig cells of the fetal and postnatal testis and in theca cells of the postnatal ovary. Here we show that male mice homozygous for a targeted deletion of the Insl3 locus exhibit bilateral cryptorchidism with free moving testes and genital ducts. These malformations are due to failure of gubernaculum development during embryogenesis. In double-mutant male mice for Insl3 and androgen receptor genes, testes are positioned adjacent to the kidneys and steadied in the abdomen by the CSL. These findings demonstrate, that the Insl3 induces gubernaculum development in an androgen-independent way, while androgen-mediated regression of the CSL occurs independently from Insl3.

Androgens↗

PubMed1998

CpG oligodeoxynucleotides trigger protective and curative Th1 responses in lethal murine leishmaniasis.

Synthetic oligodeoxynucleotides containing CpG dinucleotides (CpG-ODN) mimic the immunostimulatory qualities of bacterial DNA. We asked whether immunostimulation by CpG-ODN predisposes for a commitment toward a Th1 vs a Th2 response in Leishmania major infection, a model for a lethal Th2-driven disease, in BALB/c mice. CpG-ODN induced Th1 effector T cells in vitro and conveyed protective immunity to disease-prone BALB/c mice in vivo. Conversion to a Th1-driven resistant phenotype was associated with IL-12 production and maintained the expression of IL-12R beta2-chains. Most strikingly, CpG-ODN were even curative when given as late as 20 days after lethal L. major infection, indicating that CpG-ODN revert an established Th2 response. These findings imply an important role of bacterial DNA and CpG-ODN in the instruction of adaptive immune responses. They also point to the therapeutic potential of CpG-ODN in redirecting curative Th1 responses in Th2-driven disorders.

Animals↗

PubMed1998

Epidemiology of cutaneous melanoma in the French West Indies (Martinique).

The epidemiology of melanoma in populations of African-European descent has rarely been reported. The authors studied melanoma in the French West Indies (Martinique), where black Caribbeans and whites represented 96% and 4% of the population, respectively. Among the 85 cases of melanoma collected from 1976 to 1995, blacks represented 75% and whites, 25%. The average incidence rates were 1.48 and 0.9 per 100,000 per year in females and males, respectively. The sole of the foot represented 72% of the primary sites in blacks. Breslow's tumor thickness was > 1.5 mm in 68% of the cases. The 5-year survival was 44%.

Age Distribution↗

PubMed1998

Bacterial DNA as an evolutionary conserved ligand signalling danger of infection to immune cells.

During infection, the innate limb of the immune system senses danger (pathogens) via constitutively expressed pattern-recognition receptors, and responds with activation and secretion of pro-inflammatory cytokines. Cell-wall components of gram-positive and gram-negative bacteria, such as peptidoglycan, endotoxin or lipoteichoic acid, activate via CD14, a prototypic pattern-recognition receptor for carbohydrates. This review article focuses on an alternative recognition system of the innate immune system for the recognition of bacterial DNA. Bacterial DNA differs from eukaryotic DNA in its frequency of the dinucleotides CG and its lack of methylation. These structural differences appear to be sensed by cells of the innate immune system such as antigen-presenting cells. As a consequence bacterial DNA serves as an alternate ligand to signal danger of infection. Bacterial DNA and (synthetic) oligonucleotides (ODN) derived thereof are as efficient as endotoxin in activating macrophages and dendritic cells and in triggering release of pro-inflammatory cytokines. In mice sensitized with D-galactosamine (D-GalN), high doses of bacterial DNA from either gram-positive or gram-negative pathogens induce a lethal cytokine syndrome (lethal shock). Therefore, bacterial DNA may represent a hitherto unrecognized pathophysiological entity in host-parasite interactions. Moreover, recent evidence suggests that bacterial DNA or immunostimulating ODN triggers the immunostimulation of antigen-presenting cells, and can be utilized as adjuvant to enhance immune responses of the adaptive immune system towards poorly immunogenic antigens. In fact, foreign DNA might be useful as immunotherapeutically active adjuvant to direct adaptive immune responses towards Thl-dominated immune reactions. If these findings are operative in humans, immunostimulating ODN might be used to influence Th2-dominated diseases such as allergy.

Animals↗