Experimental studies of a coaxial plasma injector. 1 - Study of operational conditions

Parameter optimization, and operation of coaxial plasma injecto

Limits to sustained energy intake. XXIII. Does heat dissipation capacity limit the energy budget of lactating bank voles?

Acknowledgements We are grateful to our technicians and several students for their help during this study and for animal care. We thank Catherine Hambly and Peter Thompson for technical assistance for the isotope analysis for the DLW measurements. We thank Ulf Bauchinger for stimulating discussion and his comments, and two anonymous referees for comments on the manuscript. Funding This project was supported by grants from the Polish Ministry of Science and Higher Education [0595/B/P01/2011/40 to E.T.S. and 8167/B/P01/2011/40 to P.K.], and Jagiellonian University [DS/WBINOZ/INOS/757 to P.K.].Peer reviewedPublisher PD

Morphology and function of human Leydig cells in vitro. Immunocytochemical and radioimmunological analyses

The aim of our study was to show whether the cells isolated from testes of patients underwent bilateral orchiectomy for prostatic cancer are able to grown in vitro, and if so, are functionally active. Immuncytochemistry was performed to show the functional status of human cultured cells. In detail, immunolocalization of luteinizing hormone receptors (LHR), mitochondria, and cytoskeletal elements was demonstrated. Moreover, radioimmunological assay was used to measure testosterone secretion by cultured Leydig cells. Using Nomarski interference contrast and fine immunofluorescence analysis the positive immunostaining for LHR was observed in almost all Leydig cells, however it was of various intensity in individual cells. Testosterone measurement revealed significant difference between testosterone secretion by hCG-stimulated and unstimulated Leydig cells (p<0.05). Moreover, testosterone levels were significantly higher in 24- and 48-hour-cultures than in those of 72 hrs (p<0.05). Morphological analysis of Leydig cells in culture revealed the presence of mononuclear and multinucleate cells. The latter cells occurred in both hCG-stimulated and unstimulated cultures. In Leydig cells labeled with a molecular marker MitoTtracker, an abundance of mitochondria and typical distribution of microtubules and microfilaments were observed irrespective of the number of nuclei within the cell, suggesting no functional differences between mono- and multinucleate human Leydig cells in vitro. Since the percentage of multinucleate cells was similar in both hCG-stimulated and unstimulated cultures (23.70% and 22.80%), respectively, the appearance of these cell population seems to be independent of hormonal stimulation

Metro-Line Crossing Minimization: Hardness, Approximations, and Tractable Cases

Crossing minimization is one of the central problems in graph drawing. Recently, there has been an increased interest in the problem of minimizing crossings between paths in drawings of graphs. This is the metro-line crossing minimization problem (MLCM): Given an embedded graph and a set L of simple paths, called lines, order the lines on each edge so that the total number of crossings is minimized. So far, the complexity of MLCM has been an open problem. In contrast, the problem variant in which line ends must be placed in outermost position on their edges (MLCM-P) is known to be NP-hard. Our main results answer two open questions: (i) We show that MLCM is NP-hard. (ii) We give an $O(\sqrt{\log |L|})$-approximation algorithm for MLCM-P

Bean starch as ingredient for gluten-free bread

18 pages, 6 tables, 5 figures.-- Special Issue: Online Only Supplement.-- The definitive version is available at www3.interscience.wiley.comCommercially available gluten-free breads are of low quality and have a rapid staling during storage. Therefore, attempts were made to obtain gluten-free breads of improved structure properties and extended shelf life. For this purpose, laboratory-obtained bean starch, both native or hydrothermally modified, was added to a gluten-free formulation. Texture results revealed differences between the bottom (harder) and upper (softer) parts of fresh breads containing native bean starch. Modified starch reduced the hardness and diminished the differences between the upper and the bottom parts of a bread slice. Independent of storage duration, breads are crumbly. A considerable decrease of the peak and final viscosity was observed with the increase of the storage time in sample with native bean starch, whereas the presence of modified starch induced the opposite tendency. The addition of native starch increased the tendency of amylopectine to retrograde during storage, whereas the presence of modified starch decreased the retrogradation enthalpy by 16%.Peer reviewe

Experimental evolution on a wild mammal species results in modifications of gut microbial communities

Comparative studies have shown that diet, life history, and phylogeny interact to determine microbial community structure across mammalian hosts. However, these studies are often confounded by numerous factors. Selection experiments offer unique opportunities to validate conclusions and test hypotheses generated by comparative studies. We used a replicated, 15-generation selection experiment on bank voles (Myodes glareolus) that have been selected for high swim-induced aerobic metabolism, predatory behavior toward crickets, and the ability to maintain body mass on a high-fiber, herbivorous diet. We predicted that selection on host performance, mimicking adaptive radiation, would result in distinct microbial signatures. We collected foregut and cecum samples from animals that were all fed the same nutrient-rich diet and had not been subjected to any performance tests. We conducted microbial inventories of gut contents by sequencing the V4 region of the 16S rRNA gene. We found no differences in cecal microbial community structure or diversity between control lines and the aerobic or predatory lines. However, the cecal chambers of voles selected for herbivorous capability harbored distinct microbial communities that exhibited higher diversity than control lines. The foregut communities of herbivorous-selected voles were also distinct from control lines. Overall, this experiment suggests that differences in microbial communities across herbivorous mammals may be evolved, and not solely driven by current diet or other transient factors

The barriers and facilitators to the use of lifestyle apps: a systematic review of qualitative studies

Background: Mobile-health applications are revolutionising the way healthcare is being delivered. However, current research focusses on apps aimed at monitoring of conditions rather than the prevention of disease. Healthcare apps that prevent disease can be classified as lifestyle apps (LAs) and encompass mindfulness, exercise, and diet apps. In order for widespread implementation of these apps, perspectives of the user must be taken into consideration. Therefore, this systematic literature review identifies the barriers and facilitators to the use of LAs from a user’s perspective. Objective: To both identify the facilitators to the use of LAs from a user perspective as well as identify the barriers to the use of LAs from a user perspective. Methods: A systematic literature review was conducted following PRISMA guidelines. Qualitative articles focussed on a healthy non-diseased population were obtained. Two independent researchers coded the articles, and themes were identified. Results: Our results found that there were five barriers and five facilitators to app use. The facilitators included (1) motivational aspects to the user, (2) effective marketing and communication, (3) user-centred design and content, (4) humanising technology, and (5) accessibility. The five barriers identified were (1) a non-conducive, (2) poor marketing and branding, (3) controlling and invasive, (4) disengaging content, and (5) inaccessibility. Conclusions: By overcoming the barriers of LAs and encouraging the facilitators found, users are more likely to engage with this method of health promotion. Future research must be conducted on the barriers and facilitators to development and distribution of apps in order for LAs to be implemented in widespread healthcare practice

Selection of antibody single-chain variable fragments with improved carbohydrate binding by phage display.

A single-chain variable fragment (Fv) version of a murine monoclonal antibody, Se155-4, specific for Salmonella serogroup B O-polysaccharide, was used as a model system for testing monovalent phage display as a route for enhancing the relatively low affinities that typify anti-carbohydrate antibodies. Random single-chain Fv mutant libraries generated by chemical and error-prone polymerase chain reaction methods were panned against the serogroup B lipopolysaccharide. Panning of a randomly mutated heavy chain variable domain library indicated selection for improved serogroup B binders and yielded six mutants, five of which showed wild type activity by enzyme immunoassay. Two of these were apparently selected on the basis of better functional single-chain Fv yield in Escherichia coli. A heavy chain mutation (Ile77-->Thr) in one mutant, 3B1, appeared to have a particularly dramatic effect, resulting in yields of approximately 120 mg/liter of functional periplasmic product. The sixth mutant, 4B2, had complementarity determining region 1 (CDR1) and CDR2 mutations and demonstrated 10-fold improved binding, by enzyme immunoassay, relative to the wild type. Extensive analysis of antigen-antibody interactions indicated that the improved binding properties of 4B2 were attributable to a higher association rate constant and interaction with an epitope that is larger than the trisaccharide recognized by the wild type. None of the mutations involved known trisaccharide contact residues; this was consistent with analysis of wild type and mutant single-chain Fvs by titration microcalorimetry. Examination of the structure indicated that two mutations in the heavy chain CDR2 provided improved surface complementarity between the protein and the extended epitope encompassing 2 additional hexose residues. However, introduction of only the CDR2 mutations into the wild type structure failed to confer the improved binding properties of 4B2, indicating an indirect effect by the more distant mutations. Panning of randomly mutated light chain variable domain and full-length single-chain Fv mutant libraries did not yield mutants with improved assembly or binding properties

Bacterial expression and secretion of various single-chain Fv genes encoding proteins specific for a Salmonella serotype B O-antigen.

Active single-chain Fv molecules encoded by synthetic genes have been expressed and secreted to the periplasm of Escherichia coli using the ompA secretory signal. Four different constructs were developed to investigate the effects of peptide linker design and VL-VH orientation on expression, secretion, and binding to a Salmonella O-polysaccharide antigen. Peptide linker sequences derived from the elbow regions of the Fab molecule were used alone or in combination with the flexible (GGGGS)2 sequence. VL and VH domain order in the single chain molecules had a profound effect on the level of secretion but hardly influenced total expression levels, which were approximately 50 mg/liter, chiefly in the form of inclusion bodies. With VL in the NH2-terminal position, the amount of secreted product obtained was 2.4 mg/liter, but when VH occupied this position the yield was less than 5% of this value. Enzyme immunoassays of the four products showed domain order and linker sequence affected antigen binding by less than an order of magnitude. Attempts to express active Fv from dicistronic DNA were unsuccessful, but active Fv was obtained from single-chain Fv by enzymic cleavage at a site in the elbow linker peptide. The thermodynamic binding parameters of intact and cleaved single-chain Fvs determined by titration microcalorimetry were similar to those of bacterially produced Fab and mouse IgG