Mobilization of Regulatory T Cells in Response to Carotid Injury Does Not Influence Subsequent Neointima Formation

Aim

T cells have been attributed an important role in modulating repair responses following vascular injury. The aim of this study was to investigate the role of different T cell subsets in this context.

Methods and Results

A non-obstructive collar was introduced to inflict carotid artery injury in mice and subsequent activation of immune cells in draining lymph nodes and spleen were studied by flow cytometry. Carotid artery injury of wild type mice was associated with mobilization of both Th1 type CD4⁺IFNγ⁺ and regulatory CD4⁺CD25⁺FoxP3⁺ T cells in draining lymph nodes. Studies using FoxP3-green fluorescent protein (GFP) transgenic C57/Bl6 mice demonstrated scattered presence of regulatory T cells in the adventitial tissue of injured arteries as well as a massive emigration of regulatory T cells from the spleen in response to carotid injury. However, deletion of antigen presentation to CD4+ T cells (H2⁰ mice), as well as deletion of regulatory T cells (through treatment with blocking anti-CD25 antibodies), did not affect neointima formation. Also deletion of antigen presentation to CD8⁺ T cells (Tap1⁰ mice) was without effect on carotid collar-induced neointima formation.

Conclusion

The results demonstrate that carotid artery injury is associated with mobilization of regulatory T cells. Depletion of regulatory T cells does not, however, influence the subsequent repair processes leading to the formation of a neointima. The results also demonstrate that lack of CD8⁺ T cells does not influence neointima formation in presence of functional CD4⁺ T cells and B cells.     

Production of cachexia mediators by Walker 256 cells from ascitic tumors

In neoplasic cachexia, chemical mediators seem to act as initiators or perpetuators of this process. Walker 256 cells, whose metabolic properties have so far been little studied with respect to cancer cachexia, are used as a model for the study of this syndrome. The main objective of this research was to pinpoint the substances secreted by these cells that may contribute to the progression of the cachectic state. Since inflammatory mediators seem to be involved in the manifestation of this syndrome, the in vitro production of nitric oxide (NO), cytokines (tumor necrosis factor alpha (TNF-alpha) and interleukin-6 (IL-6)), and prostaglandin E2 (PGE2) was evaluated in Walker 256 cells isolated from ascitic tumors. After 4 or 5 h, a significant increase in NO production was observed (2.55 +/- 1.56 and 4.05 +/- 1.99 nmol NO per 10(7) cells, respectively). When isolated from a 6-day-old tumor, a significantly lower production of IL-6 and higher production of TNF-alpha than in cells from a 4-day-old tumor were observed, indicating a relationship between the production of cytokines and the time of tumor development after implantation. Considerable production of PGE(2) by Walker 256 cells isolated from the 6-day-old tumor was also observed. Polyamines were also determined in Walker 256 cells. Levels of putrescine, spermidine, and spermine did not show significant differences in tumors developed during 4 or 6 days. Direct evidence of the release of proinflammatory cytokines and PGE2 by Walker 256 cells suggests that these mediators can drive the cachectic syndrome in the host, the effect being dependent on tumor development time.     

Endocytosis of apolipoprotein A-V by members of the low density lipoprotein receptor and the VPS10p domain receptor families

Apolipoprotein A-V (apoA-V) is present in low amounts in plasma and has been found to modulate triacylglycerol levels in humans and in animal models. ApoA-V displays affinity for members of the low density lipoprotein receptor (LDL-R) gene family, known as the classical lipoprotein receptors, including LRP1 and SorLA/LR11. In addition to LDL-A binding repeats, the mosaic receptor SorLA/LR11 also possesses a Vps10p domain. Here we show that apoA-V also binds to sortilin, a receptor from the Vsp10p domain gene family that lacks LDL-A repeats. Binding of apoA-V to sortilin was competed by neurotensin, a ligand that binds specifically to the Vps10p domain. To investigate the biological fate of receptor-bound apoA-V, binding experiments were conducted with cultured human embryonic kidney cells transfected with either SorLA/LR11 or sortilin. Compared with nontransfected cells, apoA-V binding to SorLA/LR11- and sortilin-expressing cells was markedly enhanced. Internalization experiments, live imaging studies, and fluorescence resonance energy transfer analyses demonstrated that labeled apoA-V was rapidly internalized, co-localized with receptors in early endosomes, and followed the receptors through endosomes to the trans-Golgi network. The observed decrease of fluorescence signal intensity as a function of time during live imaging experiments suggested ligand uncoupling in endosomes with subsequent delivery to lysosomes for degradation. This interpretation was supported by experiments with (125)I-labeled apoA-V, demonstrating clear differences in degradation between transfected and nontransfected cells. We conclude that apoA-V binds to receptors possessing LDL-A repeats and Vsp10p domains and that apoA-V is internalized into cells via these receptors. This could be a mechanism by which apoA-V modulates lipoprotein metabolism in vivo.     

Sex-role reversal revisited: choosy females and ornamented, competitive males in a pipefish

In the pipefish Syngnathus typhle sex roles are reversed, thatis, females compete more intensely than males over mates. However,competition over mates among individuals of one sex does notnecessarily prevent members of that same sex from being choosy,and choosiness in the other sex does not prevent competitionwithin it. In an experiment we allowed a female pipefish tochoose freely between two males, after which we released themales and let the three interact. Comparisons with earlier resultsshow that both sexes courted partners and competed with consexuals.However, females courted more often than did males, and courtshipwas more frequent in treatments involving large individualsthan in treatments with small individuals. Males competed amongthemselves for access to mates but for a shorter duration thanfemales in the same situation. Males displayed an ornament towardsfemales but not to males during mating competition. Females,however, used their ornament in both contexts. Females did notalways mate with the male of their previously made choice, whichwe interpret as females being constrained by male-male competition,male motivation to mate, or both. Thus, in this sex-role reversedspecies, mate choice in the more competitive sex may be circumventedand even overruled by mate competition and mating willingnessin the least competitive sex. Hence, sex roles should not beconsidered as sexes being either choosy or competitive but ratherthat males and females may exhibit different combinations ofchoice and competition.     

Male pipefish prefer dominant over attractive females

Animals may obtain information guiding their choice betweenpotential partners from observing competitive interactionsand displays between them, or from displays directed at thechoosing individual. In the sex-role reversed pipefish Syngnathustyphle females display a temporary ornament (a color pattern)to other females as well as to males. We have previously shown that display of female ornaments per se is attractive to males.Here we show that information from competitive displays canoverride such direct attraction displays as signals in thepartner choice process. In a mate choice experiment, an enclosedmale could choose between two females. On the first experimentalday, females could interact freely, while on the second daythey were isolated from each other. When female-female competitionwas allowed, the ornament display was directed more to theother female than to the male: Time competing, rather thantime courting the male, correlated with ornament display duration.However, ornament display under competition and ornament displayin the absence of competition did not correlate significantly.In fact, females competing more intensively on day one displayedthe ornament less on day two. Furthermore, the ornament displayduring the first, but not the second, day predicted male matechoice on the second day. Thus, males remembered previous informationfrom competitive displays and used it rather than immediateinformation from displays in the absence of female-female competition.We suggest that competitive displays more reliably signal female quality as compared to noncompetitive ones, and that males benefitfrom mating with dominant females.     

Inhibition of prostaglandin induced uterine activity in the second trimester of pregnancy by β-mimetic adrenergic agents

As a consequence of great individual variations among pregnant women in their response to prostaglandin medication, overstimulation of the uterus might occasionally occur. The need of a drug which could safely prevent excessive and undesirable prostaglandin-induced uterine contractions is therefore apparent.Two β-mimetic adrenergic compounds, orciprenaline and ritodrine, were tested with respect to their ability to reduce prostaglandin-induced uterine activity. Both compounds have successfully depressed the intensity, as well as the frequency of uterine contractions, which were previously induced by intrauterine administration of prostaglandin F_2α. With both drugs the reduction in uterine activity was closely dose-related and accompanied by a considerable increase in the maternal heart rate. While orciprenaline did not alter the maternal blood pressure, ritodrine produced a rise in systolic, and a fall in diastolic pressure. No other side effect was recorded. When the infusion ceased, uterine activity increased in various degrees.On the basis of the present findings the possible role of prostaglandins in the physiological regulation of the uterine contractions during pregnancy is discussed.     

A Universal Method for Species Identification of Mammals Utilizing Next Generation Sequencing for the Analysis of DNA Mixtures

Species identification can be interesting in a wide range of areas, for example, in forensic applications, food monitoring and in archeology. The vast majority of existing DNA typing methods developed for species determination, mainly focuses on a single species source. There are, however, many instances where all species from mixed sources need to be determined, even when the species in minority constitutes less than 1 % of the sample. The introduction of next generation sequencing opens new possibilities for such challenging samples. In this study we present a universal deep sequencing method using 454 GS Junior sequencing of a target on the mitochondrial gene 16S rRNA. The method was designed through phylogenetic analyses of DNA reference sequences from more than 300 mammal species. Experiments were performed on artificial species-species mixture samples in order to verify the method’s robustness and its ability to detect all species within a mixture. The method was also tested on samples from authentic forensic casework. The results showed to be promising, discriminating over 99.9 % of mammal species and the ability to detect multiple donors within a mixture and also to detect minor components as low as 1 % of a mixed sample.     

A Systematic Map of Systematic Reviews in Pediatric Dentistry—What Do We Really Know?

ObjectivesTo identify, appraise and summarize existing knowledge and knowledge gaps in practice-relevant questions in pediatric dentistry.MethodsA systematic mapping of systematic reviews was undertaken for domains considered important in daily clinical practice. The literature search covered questions in the following domains: behavior management problems/dental anxiety; caries risk assessment and caries detection including radiographic technologies; prevention and non-operative treatment of caries in primary and young permanent teeth; operative treatment of caries in primary and young permanent teeth; prevention and treatment of periodontal disease; management of tooth developmental and mineralization disturbances; prevention and treatment of oral conditions in children with chronic diseases/developmental disturbances/obesity; diagnosis, prevention and treatment of dental erosion and tooth wear; treatment of traumatic injuries in primary and young permanent teeth and cost-effectiveness of these interventions. Abstracts and full text reviews were assessed independently by two reviewers and any differences were solved by consensus. AMSTAR was used to assess the risk of bias of each included systematic review. Reviews judged as having a low or moderate risk of bias were used to formulate existing knowledge and knowledge gaps.ResultsOut of 81 systematic reviews meeting the inclusion criteria, 38 were judged to have a low or moderate risk of bias. Half of them concerned caries prevention. The quality of evidence was high for a caries-preventive effect of daily use of fluoride toothpaste and moderate for fissure sealing with resin-based materials. For the rest the quality of evidence for the effects of interventions was low or very low.ConclusionThere is an urgent need for primary clinical research of good quality in most clinically-relevant domains in pediatric dentistry.     

Intraclutch Hatch Synchronization in Pheasants and Mallard Ducks

Synchronization of hatching within clutches of precocial bird species can be achieved either by acceleration or retardation, i.e. by shortening or prolonging the incubation period. The ability of mallard ( Anas platyrhynchos ) and ring-necked pheasant ( Phasianus colchicus ) embryos to accelerate or retard hatching was tested by incubating separate clutches, of which three eggs had 2 d longer or shorter incubation time than the others, and observing their individual time of pipping (breaking of the shell). Mallard embryos were able to delay hatching by on average 0.6 d (43% of the eggs delayed at least 1 d), but were better at acceleration (on average 1.3 d; 91% of the eggs accelerated more than 1 d). Conversely, pheasant embryos were only able to accelerate by 0.4 d (50% accelerated more than 1 d), but were better at delaying the hatching (1.2 days; 77% delayed more than 1 d). This difference between the species may depend on different degrees of relatedness within clutches in pheasants and mallards. It may also be an effect of the more developed sensory and neuromuscular systems in galliforms; a reduction of the incubation period would mean that the development of, for example, locomotion would be insufficient at hatching.