Histamine and H1 -histamine receptors faster venous circulation

The study has analysed the action of histamine in the rabbit venous system and evaluated its potential role in contraction during increased venous pressure. We have found that a great variety exists in histamine sensitivity and H(1) -histamine receptor expression in various types of rabbit veins. Veins of the extremities (saphenous vein, femoral vein, axillary vein) and abdomen (common iliac vein, inferior vena cava) responded to histamine by a prominent, concentration-dependent force generation, whereas great thoracic veins (subclavian vein, superior vena cavas, intrathoracic part of inferior vena cava) and a pelvic vein (external iliac vein) exhibited slight sensitivity to exogenous histamine. The lack of reactivity to histamine was not due to increased activity of nitric oxide synthase (NOS) or heme oxygenase-1. H(1) -histamine receptor expression of veins correlated well with the histamine-induced contractions. Voltage-dependent calcium channels mediated mainly the histamine-induced force generation of saphenous vein, whereas it did not act in the inferior vena cava. In contrast, the receptor-operated channels were not involved in this response in either vein. Tyrosine phosphorylation occurred markedly in response to histamine in the saphenous vein, but not in the inferior vena cava. Histamine induced a prominent ρ kinase activation in both vessels. Protein kinase C and mitogen-activated protein kinase (MAPK) were not implicated in the histamine-induced intracellular calcium sensitization. Importantly, transient clamping of the femoral vein in animals caused a short-term constriction, which was inhibited by H(1) -histamine receptor antagonist in vivo. Furthermore, a significantly greater histamine immunopositivity was detected in veins after stretching compared to the resting state. We conclude that histamine receptor density adapts to the actual requirements of the circulation, and histamine liberated by the venous wall during increased venous pressure contributes to the contraction of vessels, providing a force for the venous return.     

Maternal Condition Affects the Offspring Testosterone Concentration in Early Nestling Period: An Experimental Test on Tawny Owls

This study addressed the question of whether maternal condition during egg laying or the rearing environment has a greater effect on offspring testosterone levels. We tested this in field experiments on a population of tawny owls Strix aluco in Duna‐Ipoly National Park, Hungary. In the experiments with females of poor condition, when broods were supplied with extra food none of the nestlings died, whereas in control broods, which were not supplied with extra food, some nestlings did die. Large differences in testosterone levels were correlated positively with hatching order both in experimental and control broods. However, it was only in control broods that the later‐hatched nestlings with low testosterone concentrations, died. In the experiments with females of good condition, the males were removed and females and their broods were supplied with restricted amounts of food. In these broods starving nestlings, whose growth had stopped, were considered as having died and were removed from the nest and hand‐reared. In control broods all nestlings fledged, and both in experimental and control broods testosterone concentrations were more even between siblings. Both types of trials confirmed a maternal influence on offspring testosterone concentration: large between‐sibling differences in concentrations in the broods of females of poor condition, where some nestlings died, could not be reduced with increased food supply, and the more even concentrations in the broods of females in good condition, where all nestlings survived, could also not be increased by restricting the food supply.     

Biological availability and humic properties of dissolved organic carbon in Lake Balaton (Hungary)

The nursery role of the Mondego estuary for marine fish species was studied between June 2003 and May 2004. The spatial and temporal distribution and abundance patterns of 0-group Dicentrarchus labrax (Linnaeus, 1758), Platichthys flesus (Linnaeus, 1758) and Solea solea (Linnaeus, 1758) were analyzed based on monthly sampling surveys in five stations along the estuarine gradient. Fishing took place during the night at low water of spring tides, using a 2 m beam trawl. The spatial patterns of estuarine colonization were different according to species. D. labrax showed a wider distribution, but the main nursery ground was the same as for S. solea. Highest densities of S. solea juveniles were found in oligohaline areas, with muddy bottoms and high benthic invertebrates availability, while P. flesus occurred mainly in the sandy uppermost areas. D. labrax was found in both these areas. Fish abundance in the estuary mainly reflected seasonal changes.     

Dental anxiety in relation to emotional and behavioral problems in Croatian adolescents

The investigation was performed on 113 adolescents in the age between 15 and 18 years (63 boys, 50 girls). Corah Dental Anxiety Scale (CDAS) and Children's Fear Survey Schedule - Dental Subscale (CFSS-DS) were used for evaluation of dental fear and Child Medical Fear Questionnaire (CMFQ) for evaluation of the fear of medical treatment. Achenbach Youth Self Report questionnaire (YSR) was used for evaluation of emotional and behavioral problems. The tests were filled in by children. The aim of the study was to evaluate the level of dental anxiety in adolescents and to assess a cause--consequence relationship between dental anxiety and emotional and behavioral problems in adolescents. The results of CDAS, CFSS-DS and CMFQ tests showed that dental anxiety scores and the total internalizing problems were higher in girls. Girls displayed more physical problems (p < 0.001) and were more prone to anxiety/depression disorders (p < 0.05). Both boys and girls were more aggressive, more prone to delinquent behaviour and had more externalizing problems in comparison with the average values obtained for the Croatian population. Significant correlation coefficients for boys were calculated for age and anxiety/depression, and delinquent behaviour and aggression (p < 0.05). Significant correlations were observed between physical problems and dental anxiety measured by the CFSS-DS test (p < 0.01), and between physical problems and the total internalizing problems (p < 0.05). In girls, the CMFQ scores showed significant correlations between dental anxiety and physical problems (p < 0.05), and anxiety/ depression (p < 0.01) and the total internalizing and externalizing problems (p < 0.05). Significant correlations were calculated for age and the total internalizing and externalizing problems for boys (p < 0.05). According to the results of both CDAS and CMFQ tests, anxiety in girls showed significant correlations with delinquent behaviour (p < 0.01). CDAS scores for girls showed significant correlations with aggression (p < 0.05) and the total externalizing problems (p < 0.01).     

Yeast Rad5 protein required for postreplication repair has a DNA helicase activity specific for replication fork regression

Lesions in the template DNA strand block the progression of the replication fork. In the yeast Saccharomyces cerevisiae, replication through DNA lesions is mediated by different Rad6-Rad18-dependent means, which include translesion synthesis and a Rad5-dependent postreplicational repair pathway that repairs the discontinuities that form in the DNA synthesized from damaged templates. Although translesion synthesis is well characterized, little is known about the mechanisms that modulate Rad5-dependent postreplicational repair. Here we show that yeast Rad5 has a DNA helicase activity that is specialized for replication fork regression. On model replication fork structures, Rad5 concertedly unwinds and anneals the nascent and the parental strands without exposing extended single-stranded regions. These observations provide insight into the mechanism of postreplicational repair in which Rad5 action promotes template switching for error-free damage bypass.     

Animal models of complement-mediated hypersensitivity reactions to liposomes and other lipid-based nanoparticles

Intravenous injection of some liposomal drugs, diagnostic agents, micelles and other lipid-based nanoparticles can cause acute hypersensitivity reactions (HSRs) in a high percentage (up to 45%) of patients, with hemodynamic, respiratory and cutaneous manifestations. The phenomenon can be explained with activation of the complement (C) system on the surface of lipid particles, leading to anaphylatoxin (C5a and C3a) liberation and subsequent release reactions of mast cells, basophils and possibly other inflammatory cells in blood. These reactions can be reproduced and studied in pigs, dogs and rats, animal models which differ from each other in sensitivity and spectrum of symptoms. In the most sensitive pig model, a few miligrams of liposome (phospholipid) can cause anaphylactoid shock, characterized by pulmonary hypertension, systemic hypotension, decreased cardiac output and major cardiac arrhythmias. Pigs also display cutaneous symptoms, such as flushing and rash. The sensitivity of dogs to hemodynamic changes is close to that of pigs, but unlike pigs, dogs also react to micellar lipids (such as Cremophor EL) and their response includes pronounced blood cell and vegetative neural changes (e.g., leukopenia followed by leukocytosis, thrombocytopenia, fluid excretions). Rats are relatively insensitive inasmuch as hypotension, their most prominent response to liposomes, is induced only by one or two orders of magnitude higher phospholipid doses (based on body weight) compared to the reactogenic dose in pigs and dogs. It is suggested that the porcine and dog models are applicable for measuring and predicting the (pseudo)allergic activity of particulate "nanodrugs".     

Technology transfer from worms and flies to vertebrates: transposition-based genome manipulations and their future perspectives

To meet the increasing demand of linking sequence information to gene function in vertebrate models, genetic modifications must be introduced and their effects analyzed in an easy, controlled, and scalable manner. In the mouse, only about 10% (estimate) of all genes have been knocked out, despite continuous methodologic improvement and extensive effort. Moreover, a large proportion of inactivated genes exhibit no obvious phenotypic alterations. Thus, in order to facilitate analysis of gene function, new genetic tools and strategies are currently under development in these model organisms. Loss of function and gain of function mutagenesis screens based on transposable elements have numerous advantages because they can be applied in vivo and are therefore phenotype driven, and molecular analysis of the mutations is straightforward. At present, laboratory harnessing of transposable elements is more extensive in invertebrate models, mostly because of their earlier discovery in these organisms. Transposons have already been found to facilitate functional genetics research greatly in lower metazoan models, and have been applied most comprehensively in Drosophila. However, transposon based genetic strategies were recently established in vertebrates, and current progress in this field indicates that transposable elements will indeed serve as indispensable tools in the genetic toolkit for vertebrate models. In this review we provide an overview of transposon based genetic modification techniques used in higher and lower metazoan model organisms, and we highlight some of the important general considerations concerning genetic applications of transposon systems.     

In vitro dedifferentiation of melanocytes from adult epidermis

In previous work we described a novel culture technique using a cholera toxin and PMA-free medium (Mel-mix) for obtaining pure melanocyte cultures from human adult epidermis. In Mel-mix medium the cultured melanocytes are bipolar, unpigmented and highly proliferative. Further characterization of the cultured melanocytes revealed the disappearance of c-Kit and TRP-1 and induction of nestin expression, indicating that melanocytes dedifferentiated in this in vitro culture. Cholera toxin and PMA were able to induce c-Kit and TRP-1 protein expressions in the cells, reversing dedifferentiation. TRP-1 mRNA expression was induced in dedifferentiated melanocytes by UV-B irradiated keratinocyte supernatants, however direct UV-B irradiation of the cells resulted in further decrease of TRP-1 mRNA expression. These dedifferentiated, easily accessible cultured melanocytes provide a good model for studying melanocyte differentiation and possibly transdifferentiation. Because melanocytes in Mel-mix medium can be cultured with human serum as the only supplement, this culture system is also suitable for autologous cell transplantation.