Hemocompatibility of medical connectors with biopassive or bioactive surface coatings

Although medical connectors compose very small parts of the extracorporeal circulation (ECC) system they represent a critical localization where early thromboembolic processes can manifest. In the present study we modified an in vitro closed-loop model with fresh human whole blood for the preclinical evaluation of the hemocompatibility of three types of medical connectors: non-coated (control); with silicone-, and heparin-coating. Each single loop consists of five polycarbonate connectors joined together by five pieces of silicone tubes. Thrombin-antithrombin-III, beta-thromboglobulin (beta-TG), PMN-Elastase, terminal complement complex, CD 11b expression, and surface-absorbed fibrinogen were measured. After 1 and 2 h recirculation, platelet loss, release of beta-TG, and adsorption of fibrinogen were significantly higher (p<0.05) within the non-coated connectors compared to the silicone- and heparin-coated groups. Following this experiment, the connectors were filled again with fresh heparinized whole blood from the same donor to evaluate the influence of prior blood contact. Here, the activation of platelets and coagulation was dependent on the duration of the blood preincubation period. Probably, the coated surfaces possess a reduced, or selective adsorption of plasma proteins, which in turn leads to a faster creation of a blood-friendly secondary superficial membrane, and prevents a further denaturation and hence activation of the adsorbed proteins.     

Impaired conditioned taste aversion learning in APP transgenic mice

Cognition in transgenic mouse models of Alzheimer's disease (AD) has been predominantly characterized in explicit spatial orientation tasks. However, dementia in AD encompasses also implicit memory systems. In the present study a line of transgenic mice (TgCRND8) encoding a double mutated allele of the human amyloid precursor protein (APP) genes was evaluated in an implicit associative learning task of conditioned taste aversion (CTA). CTA is a form of Pavlovian classical conditioning, in which a mouse learns to avoid a novel taste of saccharine (conditioned stimulus) paired with an experimentally induced (systemic injection of lithium chloride) nausea (unconditioned stimulus). In contrast to conditioned non-Tg mice, TgCRND8 APP mice developed weaker aversion against saccharine and quickly increased its consumption in repeated tests. These results indicate that TgCRND8 mice show a significant impairment not only in explicit spatial memory, as has been previously shown [Nature 408 (2000) 979], but also in implicit memory. Control experiments confirmed that TgCRND8 and non-Tg mice had comparable taste sensitivities in response to appetitive as well as aversive tastes. The study suggests that the CTA paradigm can be a sensitive tool to evaluate deficits in implicit associative learning in APP transgenic mouse models of AD.     

Expression and regulation of connexins in cultured ventricular myocytes isolated from adult rat hearts

Gap junctions were assayed during re-differentiation of adult rat cardiomyocytes in long-term culture to gain insight into the processes of remodeling. Double immunostaining allowed the localization of connexins Cx40, Cx43, and Cx45 between myocytes and demonstrated co-expression and co-localization in individual cells and gap junction plaques, respectively. Immunoblots showed differential time-dependent changes in connexin expression and phosphorylation. The total amount of connexins and the ratio of phosphorylated/non-phosphorylated isoforms gradually increased during the re-establishment of intercellular communication. Dual voltage-clamp studies showed the involvement of several types of gap junction channels. Multichannel currents yielded diverse spectra of g(j,inst)=f( V(j)) and g(j,ss)=f( V(j)) relationships ( g(j,inst): instantaneous gap junction conductance; g(j,ss): conductance at steady state; V(j): transjunctional voltage), indicative of homotypic and heterotypic channels. Single-channel currents revealed two prominent conductances reflecting gamma(j,main) and gamma(j,residual). The histograms of gamma(j,main) showed four discrete peaks (41-44, 59-61, 70-76, and 100-107 pS) attributable to a combination of Cx45-Cx45, Cx40-Cx45 and Cx43-Cx45 channels (1st peak), Cx43-Cx43 and Cx40-Cx43 channels (2nd peak), Cx43-Cx43 channels (3rd peak) and Cx40-Cx40 and Cx40-Cx43 channels (4th peak). However, the presence of heteromeric channels cannot be excluded. The data are consistent with an up-regulation of Cx45 and Cx43 during re-differentiation.     

Decreased prefrontal 5-HT2A receptor binding in subjects at enhanced risk for schizophrenia

The brain serotonin-2A receptor (5-HT_2AR) has been implicated in both the pathology of schizophrenia and the therapeutic action of atypical antipsychotics. However, little is known about the 5-HT_2AR status before the onset of schizophrenia and before the exposure to antipsychotics. We used [¹⁸F] altanserin and positron emission tomography (PET) in a pilot study of 6 individuals suspected to be at elevated risk for schizophrenia and seven age-matched controls to test the hypothesis that regional 5-HT_2AR binding is altered in the prodromal stages of schizophrenia. Distribution volume ratios (DVRs) as a proxy for 5-HT_2AR availability were significantly reduced in prefrontal cortex regions of at-risk subjects, implicating early abnormalities of serotonergic neurotransmission that antecede the onset of schizophrenia.     

Die Ausbildung der Haptoglobintypen im Säuglingsalter

Zusammenfassung Das gesunde Neugeborene bildet seinen eigenen Haptoglobintyp innerhalb der 1. Lebenswoche aus. Unreife (Frühgeborene) und starke Hämolyse können das Auftreten verzögern. Am Ende des 3. Monats war der eigene Typ bei allen untersuchten Säuglingen (865) nachweisbar.     

Einige Befunde zur Sinnesphysiologie der Kaltblüterzecke Amblyomma testudinis

Zusammenfassung Aus eigener Zucht stammende Larven, Nymphen und Imagines von Amblyomma testudinis wurden auf ihre Sinnesleistungen hinsichtlich Phototaxis, Thermotaxis, Geotaxis und Chemotaxis untersucht. Es ergab sich ein entwicklungsabhängiger Wandel im phototaktischen Verhalten von anfangs positiver Phototaxis bei den Larven über indifferente Phototaxis bei vollgesogenen Larven und nüchternen Nymphen zu negativer Phototaxis bei vollgesogenen Nympen sowie - und -Imagines. Sämtliche Entwicklungsstadien mit Ausnahme der -Imagines verhalten sich temperaturindifferent vor dem Kontakt mit einem Wirt; vollgesogene Larven und Nymphen dagegen bevorzugen niedrige Temperaturen. Nymphen und Imagines, die noch nicht gesogen haben, reagieren negativ geotaktisch. Sie erklettern vermutlich in der Natur die Spitzen von Pflanzen und Steinen, um dort das Vorbeikriechen eines Wirtes abzuwarten. Imagines von Amblyomma testudinis reagieren mit einer positiven Chemotaxis auf Schlangen stärker als auf Kröten. Auch Substrat aus Schlangenbehältern enthält mindestens für 30 Std nach der Entfernung der Schlangen chemotaktisch wirksame Stoffe.

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Contributions to sensory physiology of the tick Amblyomma testudinis

Summary Phototaxis, thermotaxis, geotaxis and chemotaxis of larvae, nymphs and adults of Amblyomma testudinis from own breedings were studied. The phototactic behaviour changed according to the different stages of development: unfed larvae were positively phototactic, fed larvae and unfed nymphs were indifferent, fed nymphs but also the adults showed a negative phototaxis.—All stages of development besides the females reacted without any preference for a distinct temperature, whereas larvae and nymphs after feeding prefered lower temperatures.—Unfed nymphs and adults were negatively geotactic which corresponds to their natural behaviour of climbing plants for catching hosts.—Adults of Amblyomma testudinis showed a higher rate of chemotaxis for snakes than for toads; substrate from snake cages containes chemotaxis inducing substances for more than 30 hours after elimination of the snakes.

     

Neoglycoprotein binding to colorectal tumour cells: Comparison between primary and secondary lesions

Summary Biotinylated neoglycoproteins are useful to determine the expression of sugar receptors (lectins) histochemically in routinely processed tissue sections. Assessment of the presence of distinct receptor classes with specificity to-galactosides and to- or-N-acetylgalactosamine, selected on the basis of their potential relevance for recognition processes within the metastatic cascade in murine model systems, was performed for a common human tumour type, colorectal cancer. The four different types of neoglycoproteins, derived from covalent attachment of commercially available derivatives of-N-acetylgalactosamine, differed only quantitatively in their capacity to detect specific binding on cultured cells and tissue sections, thus posing no major restriction on the choice of synthetic process for histochemical efficiency of the product. Glycocytological application revealed specific probe binding and a regulation of level of receptor expression for a human colon carcinoma cell line primarily forN-acetylgalactosamine-specific receptors upon retinoic acid-induced differentiation. Monitoring of sections of the 12 cases of primary and secondary colorectal lesions invariably disclosed the presence of the respective receptors, the extent of cell labelling in primary tumours and metastases being similar. Establishment of metastases, even in different target organs, is apparently not followed by a major phenotypic variation in this feature.     

FISH mapping and molecular organization of the major repetitive sequences of tomato

This paper presents a bird’s-eye view of the major repeats and chromatin types of tomato. Using fluorescence in-situ hybridization (FISH) with Cot-1, Cot-10 and Cot-100 DNA as probes we mapped repetitive sequences of different complexity on pachytene complements. Cot-100 was found to cover all heterochromatin regions, and could be used to identify repeat-rich clones in BAC filter hybridization. Next we established the chromosomal locations of the tandem and dispersed repeats with respect to euchromatin, nucleolar organizer regions (NORs), heterochromatin, and centromeres. The tomato genomic repeats TGRII and TGRIII appeared to be major components of the pericentromeres, whereas the newly discovered TGRIV repeat was found mainly in the structural centromeres. The highly methylated NOR of chromosome 2 is rich in [GACA]₄, a microsatellite that also forms part of the pericentromeres, together with [GA]₈, [GATA]₄ and Ty1-copia. Based on the morphology of pachytene chromosomes and the distribution of repeats studied so far, we now propose six different chromatin classes for tomato: (1) euchromatin, (2) chromomeres, (3) distal heterochromatin and interstitial heterochromatic knobs, (4) pericentromere heterochromatin, (5) functional centromere heterochromatin and (6) nucleolar organizer region.     

Tunica media of aorta and pulmonary trunk in relation to internal calibres in transposition of great arteries,in aortic and pulmonary atresia and in normal hearts

Summary In a post mortem material of 17 cases of transposition of the great arteries (TGA) from patients with an age range from birth to two years and ten months after birth, the internal calibres of the great arteries and the ostia of the heart proved to be the same as in normal hearts. Furthermore, the media of the ascending aorta and pulmonary trunk showed no adaptation to the abnormal circulatory conditions in 15 cases of TGA with an age range from birth up to 51/2 months: in both great arteries the thickness of the tunica media and the packing density of its elastic fibres were the same as in normal hearts. However, adaptation of the tunica media of the pulmonary trunk to the abnormal circulatory conditions: increased media thickness, was found in the two remaining cases, older than 12 months.In 7 cases of pulmonary atresia (age from 1 day to 12 months) and in 9 cases of aortic atresia (age from 2 days to 37 days) the following observations were made. Vessels with reduced or absent function (ascending aorta in aortic atresia and pulmonary trunk in pulmonary atresia) showed a markedly different structure. In aortic atresia the internal calibre and thickness of the media of the ascending aorta were markedly reduced, whereas the packing density of the elastic fibres of the media remained the same as in normal hearts. In pulmonary atresia the pulmonary trunk showed large variations in internal calibre, whereas both media thickness and the packing density of its elastic fibres remained the same as in normal hearts. When the markedly enlarged single functional vessels (the pulmonary trunk in aortic atresia and the ascending aorta in pulmonary atresia) were compared no significant differences between their internal calibre, media thickness and the packing density of the elastic fibres were found indicating similar adaptation to the abnormal but comparable functional load of acting as sole arterial trunk.We are indebted to Prof. Dr. J. Moll for his help, to Dr. J.J. Willemse for statistical calculations, to Mr. P. Zondervan, M.D. (Dept. of Pathology I), for supply of material and to Mrs. L. Silvis for histo-technical assistance