Detection of a C-insertion polymorphism within the human tumor necrosis factor alpha (TNFA) gene

We have identified a C-insertion polymorphism in the 5'UTR of the first exon of the human tumor necrosis factor alpha (TNFA) gene. TNFA is a cytokine that plays an important role in the inflammatory response.     

Impact of Early Colonizers on In Vitro Subgingival Biofilm Formation

The aim of this study was to investigate the impact of early colonizing species on the structure and the composition of the bacterial community developing in a subgingival 10-species biofilm model system. The model included Streptococcus oralis, Streptococcus anginosus, Actinomycesoris, Fusobacterium nucleatum subsp. nucleatum, Veillonella dispar, Campylobacter rectus, Prevotella intermedia, Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola. Based on literature, we considered Streptococcus oralis, Streptococcus anginosus, and Actinomyces oris as early colonizers and examined their role in the biofilms by either a delayed addition to the consortium, or by not inoculating at all the biofilms with these species. We quantitatively evaluated the resulting biofilms by real-time quantitative PCR and further compared the structures using confocal laser scanning microscopy following fluorescence in situ hybridisation. The absence of the early colonizers did not hinder biofilm formation. The biofilms reached the same total counts and developed to normal thickness. However, quantitative shifts in the abundances of individual species were observed. In the absence of streptococci, the overall biofilm structure appeared looser and more dispersed. Moreover, besides a significant increase of P. intermedia and a decrease of P. gingivalis , P. intermedia appeared to form filamented long chains that resembled streptococci. A. oris, although growing to significantly higher abundance in absence of streptococci, did not have a visible impact on the biofilms. Hence, in the absence of the early colonizers, there is a pronounced effect on P. intermedia and P. gingivalis that may cause distinct shifts in the structure of the biofilm. Streptococci possibly facilitate the establishment of P. gingivalis into subgingival biofilms, while in their absence P. intermedia became more dominant and forms elongated chains.     

Collective cell migration of smooth muscle and endothelial cells: impact of injury versus non-injury stimuli

Background

Cell migration is a vital process for growth and repair. In vitro migration assays, utilized to study cell migration, often rely on physical scraping of a cell monolayer to induce cell migration. The physical act of scrape injury results in numerous factors stimulating cell migration – some injury-related, some solely due to gap creation and loss of contact inhibition. Eliminating the effects of cell injury would be useful to examine the relative contribution of injury versus other mechanisms to cell migration. Cell exclusion assays can tease out the effects of injury and have become a new avenue for migration studies. Here, we developed two simple non-injury techniques for cell exclusion: 1) a Pyrex® cylinder - for outward migration of cells and 2) a polydimethylsiloxane (PDMS) insert - for inward migration of cells. Utilizing these assays smooth muscle cells (SMCs) and human umbilical vein endothelial cells (HUVECs) migratory behavior was studied on both polystyrene and gelatin-coated surfaces.

Results

Differences in migratory behavior could be detected for both smooth muscle cells (SMCs) and endothelial cells (ECs) when utilizing injury versus non-injury assays. SMCs migrated faster than HUVECs when stimulated by injury in the scrape wound assay, with rates of 1.26 % per hour and 1.59 % per hour on polystyrene and gelatin surfaces, respectively. The fastest overall migration took place with HUVECs on a gelatin-coated surface, with the in-growth assay, at a rate of 2.05 % per hour. The slowest migration occurred with the same conditions but on a polystyrene surface at a rate of 0.33 % per hour.

Conclusion

For SMCs, injury is a dominating factor in migration when compared to the two cell exclusion assays, regardless of the surface tested: polystyrene or gelatin. In contrast, the migrating surface, namely gelatin, was a dominating factor for HUVEC migration, providing an increase in cell migration over the polystyrene surface. Overall, the cell exclusion assays - the in-growth and out-growth assays, provide a means to determine pure migratory behavior of cells in comparison to migration confounded by cell wounding and injury.

     

Tracing the genetic origin of Europe's first farmers reveals insights into their social organization

Farming was established in Central Europe by the Linearbandkeramik culture (LBK), a well-investigated archaeological horizon, which emerged in the Carpathian Basin, in today''s Hungary. However, the genetic background of the LBK genesis is yet unclear. Here we present 9 Y chromosomal and 84 mitochondrial DNA profiles from Mesolithic, Neolithic Starčevo and LBK sites (seventh/sixth millennia BC) from the Carpathian Basin and southeastern Europe. We detect genetic continuity of both maternal and paternal elements during the initial spread of agriculture, and confirm the substantial genetic impact of early southeastern European and Carpathian Basin farming cultures on Central European populations of the sixth–fourth millennia BC. Comprehensive Y chromosomal and mitochondrial DNA population genetic analyses demonstrate a clear affinity of the early farmers to the modern Near East and Caucasus, tracing the expansion from that region through southeastern Europe and the Carpathian Basin into Central Europe. However, our results also reveal contrasting patterns for male and female genetic diversity in the European Neolithic, suggesting a system of patrilineal descent and patrilocal residential rules among the early farmers.     

Human impact during the Bronze Age on the vegetation at Lago Lucone (northern Italy)

Lake-sediment records were used to reconstruct human impact on the landscape around Lago Lucone (45°33′N, 10°29′E, 249 m a.s.l.), a former lake in the western amphitheatre system of the Lago di Garda. Presence of prehistoric human populations is attested by pile-dwelling settlements from the Early-Middle Bronze Age, with one settlement at a distance of only 100 m from the coring site. Pollen, plant-macrofossil and microscopic charcoal analyses were applied to a 250 cm sediment core with four dates providing the time control. A mixed oak forest that was important during the Early-Middle Holocene was cleared and replaced by open vegetation during the Bronze Age (∼2000–1100 b.c.) when open lands were estimated to have covered more than 60% of the total relevant pollen-source area. During a phase of high human impact, independent climatic proxies suggest warm and dry climatic conditions. Later, ca. 1100 b.c., palaeobotanical evidence indicates a sharp decrease in human pressure in the Lago Lucone area. The comparison with other sedimentary palaeocultural records shows that the period 1300–1100 b.c. was characterised by general declines of agricultural activities both south and north of the Alps. These declines have been previously attributed to a change towards wetter and colder climatic conditions in and around the Alps. However, the decline in human impact around Lago Lucone cannot be exclusively attributed to climatic variation. Therefore other forcing factors independent of climatic changes, such as cultural crises or changes in spatial organisation of the habitats, cannot be ruled out under the present state of knowledge.     

The effects of surgically implanted acoustic transmitters on laboratory growth, survival and tag retention in hatchery yearling Chinook salmon

Telemetry has proven an effective means for studying the movement of fishes, however, biases associated with tagged animals requires careful scrutiny if accurate conclusions are to be made from field studies. The objective of this study was to evaluate growth, survival, and tag retention in hatchery yearling Chinook salmon (Oncorhynchus tshawytscha) juveniles with intracoelomic surgically implanted acoustic transmitters representing 2.6 to 5.6% of body weight. The first trial consisted of three treatments; passive integrated transponder (PIT) tag-only (25 fish), acoustic-tag+PIT-tag (25 fish), and sham-surgery+PIT-tag (25 fish). There were no significant differences in relative growth rate (% change in weight day^?1) among treatments over the 221 day trial. Survival in the acoustic-tag treatment (80%) was not significantly different from the PIT-tag-only and sham treatments (92 and 88% respectively). The second trial consisted of three treatments; PIT-tag-only (22 fish), acoustic-tag+PIT-tag with absorbable sutures (12 fish) and acoustic-tag+PIT-tag with non-absorbable sutures (12 fish). There were no significant differences in relative growth rate among treatments over the 160 day trial. Survival in the second trial was 100%. Fish with absorbable sutures healed sooner and with less inflammation compared to fish with non-absorbable sutures. Tag retention was 100% in both trials. The results of this study suggest that acoustic transmitters of less than 5.6% body weight can be effectively used in 1-year old Chinook salmon.