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Biomedical subjects

A Kaiser

Publications and source records attributed to A Kaiser.

At least 55 records · Page 3Linked to original sources
PubMed1999

Cloning and expression of a cDNA encoding homospermidine synthase from Senecio vulgaris (Asteraceae) in Escherichia coli.

The enzyme homospermidine synthase catalyzes the NAD+-dependent conversion of 2 mol putrescine into homospermidine. Instead of putrescine, spermidine can substitute for the first putrescine moiety in plants, in which case diaminopropane instead of ammonia is released. The enzyme facilitates the formation of the 'uncommon' polyamine homospermidine which is an important precursor in the biosynthesis of pyrrolizidine alkaloids. The first plant homospermidine synthase was purified to apparent chemical homogenity from the root tissue culture Senecio vernalis (Asteraceae) (Böttcher et al. 1994, Can. J. Chem. 72, 80-85; Ober 1997, Dissertation). Four endopeptidase LysC fragments were sequenced from the purified protein. With the aid of degenerate primers against these peptides, a cDNA encoding homospermidine synthase was now cloned and characterized from Senecio vulgaris. The nucleotide sequence of the cloned cDNA revealed an open reading frame of 1155-base pairs containing 385 amino acids with a predicted Mr of 44500. GenBank research revealed that the deduced amino acid sequence shows 59% identity to human deoxyhypusine synthase. The homospermidine synthase encoding cDNA was subcloned into the expression vector pet15b and overexpressed in E. coli. The recombinant enzyme formed upon expression catalyzed homospermidine synthesis.

Alkyl and Aryl Transferases↗
PubMed1999

Engineering of a stable whole-cell biocatalyst capable of (S)-styrene oxide formation for continuous two-liquid-phase applications.

Recombinant strains of Pseudomonas putida KT2440 carrying genetic expression cassettes with xylene oxygenase- and styrene monooxygenase-encoding genes on their chromosomes could be induced in shaking-flask experiments to specific activities that rivaled those of multicopy-plasmid-based Escherichia coli recombinants. Such strains maintained the introduced styrene oxidation activity in continuous two-liquid-phase cultures for at least 100 generations, although at a lower level than in the shaking-flask experiments. The data suggest that placement of target genes on the chromosome might be a suitable route for the construction of segregationally stable and highly active whole-cell biocatalysts.

Bacteriological Techniques↗
PubMed1999

Establishment and characterization of cytopathogenic and noncytopathogenic pestivirus replicons.

Defective interfering particles (DIs) of bovine viral diarrhea virus (BVDV) have been identified and shown to be cytopathogenic (cp) in the presence of noncytopathogenic (noncp) helper virus. Moreover, a subgenomic (sg) RNA corresponding in its genome structure to one of those BVDV DIs (DI9) was replication competent in the absence of helper virus. We report here that an sg BVDV replicon which encodes from the viral proteins only the first three amino acids of the autoprotease N(pro) in addition to nonstructural (NS) proteins NS3 to NS5B replicates autonomously and also induces lysis of its host cells. This demonstrates that the presence of a helper virus is not required for the lysis of the host cell. On the basis of two infectious BVDV cDNA clones, namely, BVDV CP7 (cp) and CP7ins- (noncp), bicistronic replicons expressing proteins NS2-3 to NS5B were established. These replicons express, in addition to the viral proteins, the reporter gene encoding beta-glucuronidase; the release of this enzyme from transfected culture cells was used to monitor cell lysis. Applying these tools, we were able to show that the replicon derived from CP7ins- does not induce cell lysis. Accordingly, neither N(pro) nor any of the structural proteins are necessary to maintain the noncp phenotype. Furthermore, these sg RNAs represent the first pair of cp and noncp replicons which mimic complete BVDV CP7 and CP7ins- with respect to cytopathogenicity. These replicons will facilitate future studies aimed at the determination of the molecular basis for the cytopathogenicity of BVDV.

Animals↗
PubMed1999

Cortisol responses to marital conflict depend on marital interaction quality.

A sample of 80 couples were videotaped discussing a marital conflict and were then grouped in 3 types according to their interaction behavior: both partners displaying predominantly negative behavior ( N = 36 couples); both partners showing positive behavior ( N = 26); and couples showing asymmetric behavior (one positive, one negative; N = 16). Positive o r negative in this context refers to the empirically defined quality of speaker and listener skills by the Kategoriensystem Partnerschaftlicher Interaktion. Psychophysiological responses were measured 5 times, both before and after the conflict discussion. Participants rated their overall marital quality, the number of marital problems, and their actual cognitions and emotions. Overall responses to the conflict revealed a greater cortisol response in women than in men. Couples grouped according to their interaction style showed significant differences in cortisol responses: In couples with positive interaction, cortisol increased markedly, whereas couples with negative interaction showed a nonresponse, that is, a slight decrease expected with diurnal variation, although they rated the actual conflict discussion as "stressful" like the other groups of couples. We conclude that marital interaction directly affects physiological responses to a conflict depending on interaction quality.

Journal Article↗
PubMed1999

A perception experiment with time-critical graphics animation on the World-Wide Web.

The World-Wide Web offers a potentially interesting tool to collect data from a large and heterogeneous audience. While questionnaires have become rather common on the Internet, its potential reaches far beyond text processing. In principle, it is possible not only to perform interactive, dynamic experiments on the Web, but also to include graphical animation and time-critical responses, such as reaction times. We implemented a visual motion extrapolation task on the Web using the programming language Java, which can be interpreted by standard Web browsers such as Netscape or Internet Explorer. The data collected with this method turned out to be reliable and differed little from data obtained in a controlled laboratory setting, with the exception of conditions with fixation instruction. Thus, the Web can, generally speaking, be used for data collection of large sample sizes. The strengths and weaknesses of dynamic visual simulation experiments on the Internet are discussed.

Adult↗
PubMed1999

Motor neuron degeneration after sciatic nerve avulsion in adult rat evolves with oxidative stress and is apoptosis.

The mechanisms for motor neuron degeneration and regeneration in adult spinal cord following axotomy and target deprivation are not fully understood. We used a unilateral sciatic nerve avulsion model in adult rats to test the hypothesis that retrograde degeneration of motor neurons resembles apoptosis. By 21 days postlesion, the number of large motor neurons in lumbar spinal cord was reduced by approximately 30%. The death of motor neurons was confirmed using the terminal transferase-mediated deoxyuridine triphosphate-biotin nick-end labeling method for detecting fragmentation of nuclear DNA. Motor neuron degeneration was characterized by aberrant accumulation of perikaryal phosphorylated neurofilaments. Structurally, motor neuron death was apoptosis. Apoptotic motor neurons undergo chromatolysis followed by progressive cytoplasmic and nuclear condensation with chromatin compaction into uniformly large round clumps. Prior to apoptosis, functionally active mitochondria accumulate within chromatolytic motor neurons, as determined by cytochrome c oxidase activity. These dying motor neurons sustain oxidative damage to proteins and nucleic acids within the first 7 days after injury during the progression of apoptosis, as identified by immunodetection of nitrotyrosine and hydroxyl-modified deoxyguanosine and guanosine. We conclude that the retrograde death of motor neurons in the adult spinal cord after sciatic nerve avulsion is apoptosis. Accumulation of active mitochondria within the perikaryon and oxidative damage to nucleic acids and proteins may contribute to the mechanisms for apoptosis of motor neurons in the adult spinal cord.

Animals↗
PubMed1998

Retinoic acid receptor gamma1 expression determines retinoid sensitivity in pancreatic carcinoma cells.

BACKGROUND & AIMS: Retinoids inhibit growth and induce differentiation in a variety of pancreatic carcinoma cells. The goal of this study was to examine the molecular mechanisms responsible for retinoid sensitivity. METHODS: Anchorage-independent growth was examined in AR42J, DSL-6A/C1, and Capan-2 cells using a human tumor clonogenic assay. Retinoid receptors were characterized by a reverse-transcription polymerase chain reaction. Retinoic acid receptor gamma1 (RARgamma1) was stably transfected into AR42J cells using lipofectamin and into DSL-6A/C1 using ballistomagnetic gene transfer. Receptor expression was verified using Southern and Northern blotting as well as electrophoretic mobility shift assays. RESULTS: Retinoid treatment resulted in a dose-dependent growth inhibition of Capan-2 cells, whereas growth was not affected in AR42J and DSL-6A/C1 cells. A selective loss of RARgamma1 expression was observed in both retinoid-resistant cell lines, whereas all other retinoid receptor subtypes showed an identical expression pattern. Retinoid treatment of three independent RARgamma1-expressing cell clones of AR42J and DSL-6A/C1 cells resulted in pronounced growth inhibition compared with wild-type control cells. CONCLUSIONS: RARgamma1 expression determines sensitivity of pancreatic carcinoma cells to retinoid-mediated growth inhibition and might therefore serve as a valuable predictive marker for retinoid treatment of pancreatic cancer.

Animals↗
PubMed1998

The efficacy of a compact psychoeducational group training program for married couples.

There is a lack of psychoeducational programs for married couples who, although subjectively concerned about their marriages, do not seek marital therapy. In this study, the efficacy of a cognitive-behavioral program delivered on a weekend by 2 trainers for groups of 4 couples was investigated. Couples (n = 67) were recruited by newspaper announcements, then randomly assigned to the intervention program or a waiting-list control group. The program consisted of communication and problem-solving training, couples' discussions to clarify their relationship expectations, and exercises to enhance their sensual or sexual relationship. At postassessment, intervention couples emitted more positive verbal and nonverbal communication behaviors during a conflict discussion task than did control couples, who reported significantly more relationship problem areas and displayed more negative communication behaviors. At the 1-year follow-up, intervention couples reported fewer problem areas in comparison with preassessment.

Adult↗
PubMed1998

Retinoic acid receptor alpha mediates growth inhibition by retinoids in rat pancreatic carcinoma DSL-6A/C1 cells.

During carcinogenesis, pancreatic acinar cells can dedifferentiate into ductal adenocarcinoma of the pancreas. DSL-6A/C1 cells represent an in vitro model of this carcinogenic sequence. This study was designed to examine the effects of retinoids on cell growth in DSL-6A/C1 cells and to characterize further the molecular mechanisms underlying the antiproliferative actions of retinoids. Treatment of DSL-6A/C1 cells with retinoids results in a time- and dose-dependent inhibition of cell growth, paralleled by a retinoid-mediated transactivation of a pTK::betaRAREx2-luciferase reporter construct transiently transfected into DSL-6A/C1 cells. Retinoid receptor expression was evaluated by reverse transcriptase polymerase chain reaction (RT-PCR) using subtype-specific primers and demonstrated expression of retinoic acid receptor alpha (RAR-alpha), RAR-beta and retinoid X receptor alpha (RXR-alpha). Using a panel of receptor subtype-specific agonists, the RAR-alpha specific agonist Ro 40-6055 was the most potent retinoid in terms of growth inhibition. Furthermore, all-trans-retinoic acid-mediated growth inhibition and transactivation was completely blocked by the RAR-alpha-specific antagonist Ro 41-5253. In summary, the RAR-alpha subtype predominantly mediates the antiproliferative effects of retinoids in DSL-6A/C1 cells. Furthermore, this cell system provides a feasible tool to study the molecular mechanisms underlying the growth inhibitory effects of retinoids in ductal pancreatic carcinoma cells derived from a primary acinar cell phenotype.

Animals↗
PubMed1997

A fluorescent substrate of transglutaminase for detection and characterization of glutamine acceptor compounds.

A fluorescent dipeptide was designed to discover glutamine acceptor proteins of transglutaminase. Starting materials for synthesis were the commercially available compounds carbobenzoxy-L-glutaminylglycine (CBZ-Gln-Gly) and monodansylcadaverine (C-DNS) which were coupled to obtain CBZ-Gln-Gly-C-DNS 1 [1-N-(carbobenzoxy-L-glutaminylglycyl)-5-N- (5'-N', N'-dimethylamino-1'-naphthalenesulfonyl)- diamidopentane]. The glutamine peptide is a substrate of bacterial transglutaminase from Streptoverticillium mobaraense as well as of the guinea pig liver enzyme. This could be shown by incorporating 1 into alpha s1-casein resulting in a significant increase in fluorescence intensity and a concomitant inhibition of cross-linking reaction. Additionally, dipeptide 1 is a useful tool to characterize the specificity of transglutaminase toward small primary amines. We established a sensitive HPLC assay and determined the kinetic parameters of several alkylamines. Hydrolysis of 1 is suppressed in the presence of the nucleophiles as it could be demonstrated with different concentrations of butylamine in semiquantitative studies. Together with labeled primary amines, reagent 1 seems to be a particularly suitable tool for examining acceptor-donor relationships of transglutaminase substrates.

Dipeptides↗
PubMed1997

Factors associated with work resumption: a 5 year follow-up with cardiac patients.

Physiological and psychological parameters of 41 cardiac patients who retired after an inpatient rehabilitation treatment in Germany were compared with those of 41 patients (matched exactly according to sex, age, and diagnosis) who worked after a 5 year follow-up. Both samples were selected from a large pool of 733 consecutive cardiac patients participating in the rehabilitation program with follow-ups at 1, 3 and 5 years. The following physiological parameters were assessed: clinical characteristics such as biochemical variables and functional parameters of the left ventricle, performance and electrocardiogram during ergometric exercise, and medication. Psychological parameters assessed with questionnaires comprised: life satisfaction, physical complaints, and illness behavior, socioeconomic and anamnestic data. No substantial differences were observed for medical data, but psychological parameters showed striking differences. Retired patients were characterized by lower work satisfaction, complaints of being more handicapped by the disease, higher 'propensity for pension', more frequent complaints concerning their general state of health, and lower education level.

Alcohol Drinking↗
PubMed1997

Rapid stimulation of a soybean protein-serine kinase that phosphorylates a novel bZIP DNA-binding protein, G/HBF-1, during the induction of early transcription-dependent defenses.

The G-box (CACGTG) and H-box (CCTACC) cis elements function in the activation of phenylpropanoid biosynthetic genes involved in the elaboration of lignin precursors, phytoalexins and the secondary signal salicylic acid as early responses to pathogen attack. We have isolated a soybean cDNA encoding a novel bZIP protein, G/HBF-1, which binds to both the G-box and adjacent H-box in the proximal region of the chalcone synthase chs15 promoter. While G/HBF-1 transcript and protein levels do not increase during the induction of phenylpropanoid biosynthetic genes, G/HBF-1 is phosphorylated rapidly in elicited soybean cells, almost exclusively on serine residues. Using recombinant G/HBF-1 as a substrate, we identified a cytosolic protein-serine kinase that is rapidly and transiently stimulated in cells elicited with either glutathione or an avirulent strain of the soybean pathogen Pseudomonas syringae pv. glycinea. Phosphorylation of G/HBF-1 in vitro enhances binding to the chs15 promoter and we conclude that stimulation of G/HBF-1 kinase activity and G/HBF-1 phosphorylation are terminal events in a signal pathway for activation of early transcription-dependent plant defense responses.

Amino Acid Sequence↗
PubMed1997

Retinoic acid receptor beta regulates growth and differentiation in human pancreatic carcinoma cells.

BACKGROUND & AIMS: Retinoic acid receptor beta (RAR beta) expression is lost or decreased during malignant transformation in human pancreatic adenocarcinoma. The aim of this study was to evaluate the role of RAR beta expression in the propagation of a malignant phenotype in human pancreatic carcinoma cells. METHODS: Overexpression of RAR beta in the human pancreatic carcinoma cell line DAN-G was achieved by selecting stable transfected cell clones. Genomic integration and expression were verified by Southern and Northern blotting and electrophoretic mobility shift assays. Growth was determined by cell number and xenografts transplanted into nude mice. Differentiation was examined by immunohistochemistry. RESULTS: Overexpression of RAR beta in DAN-G cells inhibited cellular proliferation in vitro and in vivo. Furthermore, RAR beta overexpression resulted in induction of cellular differentiation in xenografted tumors as evidenced by increased tumor cell expression of duct cell differentiation markers carcinoembryonic antigen (CEA), CA19-9, and cytokeratin 7. CONCLUSIONS: Decreased expression of RAR beta plays a key role in the maintenance of a malignant phenotype in human pancreatic adenocarcinoma and therefore represents a novel target for experimental strategies in the treatment of pancreatic cancer patients.

Animals↗
PubMed1997

Retinoids inhibit adhesion to laminin in human pancreatic carcinoma cells via the alpha 6 beta 1-integrin receptor.

BACKGROUND & AIMS: The initial step in tumor invasion and metastasis is determined by adhesion of tumor cells to basement membranes. To evaluate their potential therapeutic use in controlling local growth and metastasis, the effects of retinoids on the adhesive properties in the human pancreatic carcinoma cell line DAN-G were examined. METHODS: The effects of retinoids on cellular adhesion were assessed by adhesion assays in vitro. The expression of laminin-binding proteins was characterized by Northern blotting, radioimmunoprecipitation, and flow-cytometric analysis. RESULTS: Treatment with retinoids results in a time- and dose-dependent inhibition of DAN-G cell adhesion to fibronection and laminin but not to collagens I, IV, and VI. The adhesion of DAN-G cells to laminin could be blocked completely by anti-alpha 6 and anti-beta 1 antibodies but not by the synthetic peptide YIGSR. Flow-cytometric analysis of DAN-G cells showed no quantitative difference for alpha 6-integrin expression in retinoid-treated and -untreated DAN-G cells. Furthermore, radioimmunoprecipitation showed no difference in the appearance of alpha 6 beta 1-integrin expression after retinoid incubation. CONCLUSIONS: Retinoids decrease pancreatic carcinoma cell adhesion to laminin via an as yet unidentified mechanism involving alteration of the alpha 6 beta 1-integrin receptor function and thereby open interesting perspectives for the modulation of infiltrative growth and metastasis in pancreatic cancer.

Animals↗
PubMed1997

Dispersal behavior of adult snow melt mosquitoes in the Upper Rhine Valley, Germany.

The dispersal behavior of female snow melt mosquitoes was studied in two forests in Rhineland-Palatinate, Germany, from April to August 1993. Both CDC-light-traps and human bait collections were used to collect mosquitoes. Sampling sites were chosen along a west-east and a north-south transect in treated and untreated parts of a forest with a village in its center. Around this settlement, breeding sites within a radius of 1.5 to 2.5 km were treated. It could be shown that this buffer zone is sufficient to prevent a nuisance caused by snow melt mosquitoes in the village. The results lead to the conclusion that snow melt mosquitoes do not regularly migrate over large distances but stay near their breeding sites. In a detailed study of the behavior of Aedes rusticus, it could be observed that these mosquitoes were resting in the interior of the forest during daytime and leaving it with increasing dusk up to 50 m from the forest edge. A comparison of landing rate counts near a row of trees and in the open field showed higher activity near the row of trees indicating visual orientation of the mosquitoes. Although the Ae. rusticus females left the forest regularly, no nuisance occurred in nearby villages. The treatment of breeding sites near settlements appeared to be sufficient to prevent a nuisance caused by the snow melt mosquitoes.

Aedes↗
PubMed1996

Brainstem connections of the macula lagenae in the chicken.

The macula lagenae, an otolithic hair-cell organ with probable vestibular function, lies close to the apical end of the avian auditory hair-cell epithelium, the papilla basilaris. In an earlier study in the pigeon in which lesioning techniques were used, Boord and Rasmussen ([1963] J. Comp. Neurol. 120:463-473) reported finding a projection of lagenar fibers to parts of the cochlear nuclei (nucleus magnocellularis and nucleus angularis). Subsequent to this report, it has been generally assumed that at least part of the cochlear nuclei has a vestibular or a combined vestibular-auditory function. In this study, we labeled fibers innervating the macula lagenae of the chicken by using a lipophilic fluorescent tracer. The analysis of Vibratome sections of the brainstem with epifluorescence illumination showed no projection to the cochlear nuclei. In cases in which the apical part of the papilla basilaris was contaminated with tracer, however, we found labeling of the cochlear nuclei in the same areas as described with the lesioning technique in the pigeon. Our results thus imply that there is no processing of information from the macula lagenae in the cochlear nucleus of the chicken. In addition, we studied the origin of the few labeled efferent neurons in the brainstem. The location of all somata encountered was restricted to an area medial to the nucleus facialis dorsalis and corresponded to the dorsal efferent cell group, from which efferents to other vestibular organs also originate.

Afferent Pathways↗