Analysis and three-dimensional visualization of collagen in artificial scaffolds using nonlinear microscopy techniques

Extracellularly distributed collagen and chondro- cytes seeded in gelatine and poly-e-caprolactone scaffolds are visualized by two-photon excitation microscopy (TPEM) and second-harmonic generation (SHG) imaging in both for- ward and backward nondescanned modes. Joint application of TPEM and SHG imaging in combination with stereolog- ical measurements of collagen enables us not only to take high-resolution 3-D images, but also to quantitatively an- alyze the collagen volume and a spatial arrangement of cell-collagen-scaffold systems, which was previously im- possible. This novel approach represents a powerful tool for the analysis of collagen-containing scaffolds with applica- tions in cartilage tissue engineering. C 2010 Society of Photo-Optica

Thermus and the Pink Discoloration Defect in Cheese

peer-reviewedA DNA sequencing-based strategy was applied to study the microbiology of Continental-type cheeses with a pink discoloration defect. The basis for this phenomenon has remained elusive, despite decades of research. The bacterial composition of cheese containing the defect was compared to that of control cheese using 16S rRNA gene and shotgun metagenomic sequencing as well as quantitative PCR (qPCR). Throughout, it was apparent that Thermus, a carotenoid-producing genus, was present at higher levels in defect-associated cheeses than in control cheeses. Prompted by this finding and data confirming the pink discoloration to be associated with the presence of a carotenoid, a culture-based approach was employed, and Thermus thermophilus was successfully cultured from defect-containing cheeses. The link between Thermus and the pinking phenomenon was then established through the cheese defect equivalent of Koch’s postulates when the defect was recreated by the reintroduction of a T. thermophilus isolate to a test cheese during the manufacturing process. IMPORTANCE Pink discoloration in cheese is a defect affecting many cheeses throughout the world, leading to significant financial loss for the dairy industry. Despite decades of research, the cause of this defect has remained elusive. The advent of high-throughput, next-generation sequencing has revolutionized the field of food microbiology and, with respect to this study, provided a means of testing a possible microbial basis for this defect. In this study, a combined 16S rRNA, whole-genome sequencing, and quantitative PCR approach was taken. This resulted in the identification of Thermus, a carotenoid-producing thermophile, in defect-associated cheeses and the recreation of the problem in cheeses to which Thermus was added. This finding has the potential to lead to new strategies to eliminate this defect, and our method represents an approach that can be employed to investigate the role of microbes in other food defects of unknown origin.Teagasc Walsh Fellowship Programm

Enhanced Growth and Osteogenic Differentiation of Human Osteoblast-Like Cells on Boron-Doped Nanocrystalline Diamond Thin Films

Intrinsic nanocrystalline diamond (NCD) films have been proven to be promising substrates for the adhesion, growth and osteogenic differentiation of bone-derived cells. To understand the role of various degrees of doping (semiconducting to metallic-like), the NCD films were deposited on silicon substrates by a microwave plasma-enhanced CVD process and their boron doping was achieved by adding trimethylboron to the CH4:H2 gas mixture, the B∶C ratio was 133, 1000 and 6700 ppm. The room temperature electrical resistivity of the films decreased from >10 MΩ (undoped films) to 55 kΩ, 0.6 kΩ, and 0.3 kΩ (doped films with 133, 1000 and 6700 ppm of B, respectively). The increase in the number of human osteoblast-like MG 63 cells in 7-day-old cultures on NCD films was most apparent on the NCD films doped with 133 and 1000 ppm of B (153,000±14,000 and 152,000±10,000 cells/cm2, respectively, compared to 113,000±10,000 cells/cm2 on undoped NCD films). As measured by ELISA per mg of total protein, the cells on NCD with 133 and 1000 ppm of B also contained the highest concentrations of collagen I and alkaline phosphatase, respectively. On the NCD films with 6700 ppm of B, the cells contained the highest concentration of focal adhesion protein vinculin, and the highest amount of collagen I was adsorbed. The concentration of osteocalcin also increased with increasing level of B doping. The cell viability on all tested NCD films was almost 100%. Measurements of the concentration of ICAM-1, i.e. an immunoglobuline adhesion molecule binding inflammatory cells, suggested that the cells on the NCD films did not undergo significant immune activation. Thus, the potential of NCD films for bone tissue regeneration can be further enhanced and tailored by B doping and that B doping up to metallic-like levels is not detrimental for cells

Eastern European women in Akureyri

Innflytjendum hefur fjölgað á Íslandi síðustu tvo áratugi, sérstaklega frá Austur- Evrópu. Þess vegna hafa nokkrar stefnur varðandi aðlögun innflytjenda verið kynntar. Þær leggja áherslu á íslenska tungu og atvinnuþátttöku. Flestir innflytjendur koma til Íslands til að vinna og dvöl þeirra er oft álitin tímabundin bæði af innfæddum og innflytjendunum sjálfum. Samþætting hefur jákvæð áhrif fyrir innflytjendur en er aðeins möguleg þegar bæði innflytjendur og innfæddir samþykkja hver annan. Þessi rannsókn skoðar aðlögun Austur-Evrópskra innflytjendakvenna, þar sem skoðað er hvort breyting sé á sjónarmiðum þeirra. Rætt var tvisvar við þær, fyrst árið 2017 og síðan árið 2020. Sjálfsálit þátttakenda var metið til að sjá hvernig það hafði áhrif á líf þeirra. Niðurstöðurnar sýndu litlar breytingar á þessum tíma, jafnvel þó að búist hefði verið við meiri breytingum. Íslenskukunnátta breytti ekki öllu þó að flestir telji að tungumálið sé lykilatriði, betri færni í tungumálinu er sjaldan tengd góðri atvinnu, launum eða almennri ánægju. Þeir sem höfðu lélega íslenskukunnáttu náðu ekki framförum á þessum þrem árum, jafnvel þegar þeim fannst þeir eiga að hafa gert það. Hámenntaðir þátttakendur notuðu sjaldan menntun sína í starfi. Þeir sem voru sjálfstætt starfandi sýndu mesta ánægju með störf þeirra. Félagsleg tengsl milli þátttakenda og Íslendinga voru tiltölulega léleg, aðeins fáir áttu íslenska vini. Ekki sýndu margir stjórnmálum áhuga eða vilja til að gerast íslenskur ríkisborgari. Skammtíma áætlarnir höfðu neikvæð áhrif á ákvarðanir þátttakenda um að læra tungumálið, bæta stöðu þeirra á vinnumarkaði eða stækka tengslanet. Marga skorti hvata til að stíga skref í átt að breytingum. Lítið sjálfsálit hafði neikvæð áhrif á suma þátttakendur og lítillækkandi hegðunarmynstur kom í ljós. Þeir þátttakendur sem höfðu aðlagast best töldu að hlutdrægar skoðanir væru gagnvart innflytjendum frá Austur-Evrópu og vildu ekki einbeita sér að uppruna þeirra.The number of immigrants, especially from Eastern Europe, has been increasing in Iceland over the last two decades. Consequently, several immigrant integration policies have been presented. They emphasise the Icelandic language and labour market participation. Most immigrants come to Iceland as labour migrants, and their stay is often perceived to be temporary both by the native population and the immigrants themselves. Integration supports positive outcomes for immigrants but is possible only when both the immigrants and the native population accept each other. This study examines Eastern European immigrant women’s integration in two waves of interviews, first in 2017 and then in 2020, inquiring whether there is a change over time in their perspectives. The self-esteem of the participants was assessed to see how it influences their migration experiences. The results showed minimal alterations over time, even if a change was anticipated. The Icelandic language plays an ambiguous role. While most believed fluency of the language is essential, better skills rarely correlated with suitable employment, pay or overall satisfaction. Those who had poor Icelandic skills did not make noticeable improvements three years later while initially believing they would. The highly educated participants rarely used their education, and those who were self-employed showed most satisfaction with their jobs. Social connections between the participants and Icelanders were relatively poor, only a few had Icelandic friends. Not many showed interest in politics or becoming an Icelandic citizen. A common pattern of short-term plans negatively influenced participants’ decisions on learning the language, improving their position on the labour market, and expanding networks. Many lacked the motivation to take steps towards change. Low self-esteem negatively influenced some participants, and self-defeating behaviour patterns became apparent. The participants who seemed most integrated felt biased views towards Eastern European immigrants, making them unwilling to focus on their origin.Byggðastofnu

Underemployment of Immigrant Women in Iceland – A case study

The number of immigrants living in Iceland has been steadily on the rise for the last decade; between 2007 and 2017, the percentage of immigrants living in Iceland has increased from 7.6 % to 11.9%. Akureyri, the largest town in the North of Iceland with considerable industry and service, has seen its immigrant population double in the last decade, and is now home to 931 immigrants for a total of 18 488 inhabitants. New research from the University of Akureyri[1]shows that immigrant women are the most vulnerable people in the labour market in Iceland. Many occupy positions that do not fit with their level of education; despite having received higher education than men. For example, in the survey conducted 30% of immigrant women in Akureyri answered that they are in employment that does not suit their background, compared to the same answer by only 8% of Icelandic women. This difference has a direct impact on the income: just 11% of immigrant women answered that they earn 300 000 ISK or more per month, compared to 37% for Icelandic women and 22% for immigrant men

Hidden in Plain Sight: Comprehensive Molecular Phylogeny of Keroplatidae and Lygistorrhinidae (Diptera) Reveals Parallel Evolution and Leads to a Revised Family Classification

We provide the first molecular phylogeny of Keroplatidae and Lygistorrhinidae, families of fungus gnats (Diptera: Bibionomorpha: Sciaroidea). Phylogenies reconstructed by Maximum Likelihood and Bayesian methods, based on four nuclear and four mitochondrial gene markers (5106 base pairs) sequenced for 75 genera and 105 species, show Keroplatidae as monophyletic only with the family Lygistorrhinidae included, herewith treated as the subfamily Lygistorrhininae stat. nov. The subfamily Arachnocampinae is retained in the family, although lowering its overall support. An early branching clade, comprising species of Platyura Meigen, 1803 and Paleoplatyura melanderi Fisher, 1941, forms subfamily Platyurinae Loew, 1850 stat. nov. The subfamilies Sciarokeroplatinae and Macrocerinae grouped together with three genera considered here as Keroplatidae incertae sedis. Subfamily Lygistorrhininae forms a sister clade to subfamily Keroplatinae, both retained monophyletic with high support. The traditional division of the subfamily Keroplatinae into the tribes Orfeliini and Keroplatini appears as outdated, resting largely on adaptive characters prone to parallel evolution. We find support for an alternative tribe corresponding to the Cloeophoromyia–Asindulum genus group, but a tribal reclassification of the Keroplatinae is left for future studies. The genus Heteropterna Skuse, 1888 is considered as identical with Ctenoceridion Matile, 1972 syn. nov

Osteogenic cell differentiation on H-terminated and O-terminated nanocrystalline diamond films

Jana Liskova,1 Oleg Babchenko,2 Marian Varga,2 Alexander Kromka,2 Daniel Hadraba,1 Zdenek Svindrych,1 Zuzana Burdikova,1 Lucie Bacakova1 1Institute of Physiology, Academy of Sciences of the Czech Republic, Prague, Czech Republic; 2Institute of Physics, Academy of Sciences of the Czech Republic, Prague, Czech Republic Abstract: Nanocrystalline diamond (NCD) films are promising materials for bone implant coatings because of their biocompatibility, chemical resistance, and mechanical hardness. Moreover, NCD wettability can be tailored by grafting specific atoms. The NCD films used in this study were grown on silicon substrates by microwave plasma-enhanced chemical vapor deposition and grafted by hydrogen atoms (H-termination) or oxygen atoms (O-termination). Human osteoblast-like Saos-2 cells were used for biological studies on H-terminated and O-terminated NCD films. The adhesion, growth, and subsequent differentiation of the osteoblasts on NCD films were examined, and the extracellular matrix production and composition were quantified. The osteoblasts that had been cultivated on the O-terminated NCD films exhibited a higher growth rate than those grown on the H-terminated NCD films. The mature collagen fibers were detected in Saos-2 cells on both the H-terminated and O-terminated NCD films; however, the quantity of total collagen in the extracellular matrix was higher on the O-terminated NCD films, as were the amounts of calcium deposition and alkaline phosphatase activity. Nevertheless, the expression of genes for osteogenic markers – type I collagen, alkaline phosphatase, and osteocalcin – was either comparable on the H-terminated and O-terminated films or even lower on the O-terminated films. In conclusion, the higher wettability of the O-terminated NCD films is promising for adhesion and growth of osteoblasts. In addition, the O-terminated surface also seems to support the deposition of extracellular matrix proteins and extracellular matrix mineralization, and this is promising for better osteoconductivity of potential bone implant coatings. Keywords: nanocrystalline diamond film, osteoblast, Saos-2, collagen, SH

Measurement of pH micro-heterogeneity in natural cheese matrices by flourescence lifetime imaging

Cheese, a product of microbial fermentation may be defined as a protein matrix entrapping fat, moisture, minerals and solutes as well as dispersed bacterial colonies. The growth and physiology of bacterial cells in these colonies may be influenced by the microenvironment around the colony, or alternatively the cells within the colony may modify the microenvironment (e.g., pH, redox potential) due to their metabolic activity. While cheese pH may be measured at macro level there remains a significant knowledge gap relating to the degree of micro-heterogeneity of pH within the cheese matrix and its relationship with microbial, enzymatic and physiochemical parameters and ultimately with cheese quality, consistency and ripening patterns. The pH of cheese samples was monitored both at macroscopic scale and at microscopic scale, using a non-destructive microscopic technique employing C-SNARF-4 and Oregon Green 488 fluorescent probes. The objectives of this work were to evaluate the suitability of these dyes for microscale pH measurements in natural cheese matrices and to enhance the sensitivity and extend the useful pH range of these probes using fluorescence lifetime imaging (FLIM). In particular, fluorescence lifetime of Oregon Green 488 proved to be sensitive probe to map pH micro heterogeneity within cheese matrices. Good agreement was observed between macroscopic scale pH measurement by FLIM and by traditional pH methods, but in addition considerable localized microheterogeneity in pH was evident within the curd matrix with pH range between 4.0 and 5.5. This technique provides significant potential to further investigate the relationship between cheese matrix physico-chemistry and bacterial metabolism during cheese manufacture and ripening