Dynamics of gene expression during bone matrix formation in osteogenic cultures derived from human embryonic stem cells in vitro

Characterization of directed differentiation protocols is a prerequisite for understanding embryonic stem cell behavior, as they represent an important source for cell-based regenerative therapies. Studies have investigated the osteogenic potential of human embryonic stem cells (HESCs), building upon those using pre-osteoblastic cells, however no consensus exists as to whether differentiating HESCs behave in a similar manner to the traditionally used osteoblastic progenitors. Thus, the aim of the current investigation was to define the gene expression pattern of osteoblastic differentiating HESCs, treated with ascorbic acid phosphate, β-glycerophosphate and dexamethasone over a 25 day period. Characterization of the gene expression dynamics revealed a phasic pattern of bone-associated protein synthesis. Collagen type I and osteopontin were initially expressed in proliferating immature cells, whereas osterix was up-regulated at the end of active cellular proliferation. Subsequently, mineralization-associated proteins, bone sialoprotein and osteocalcin were detected. In light of this dynamic expression pattern, we concluded that two distinguishable phases occurred during osteogenic HESC differentiation; first, cellular proliferation and secretion of a pre-maturational matrix, and second the appearance of osteoprogenitors with characteristic extracellular matrix synthesis. Establishment of this model provided the foundation of a time-frame for the additional supplementation with growth factors, BMP2 and VEGF. BMP2 induced the expression of principle osteogenic factors, such as osterix, bone sialoprotein and osteocalcin, whereas VEGF had the converse effect on the gene expression pattern.     

Tadpole fingerprinting: Using tail venation patterns to photo-identify tadpole individuals of a threatened frog

Traditional methods for identifying individual amphibians in capture–mark–recapture (CMR) studies have been primarily confined to post-metamorphic stages, using artificial markers that come with a variety of limitations. An alternative that may open CMR studies to earlier life stages involves the use of a species' natural external markers in photo-based identification. In this study, we investigated whether it was possible to distinguish tadpoles of the threatened green and golden bell frog (Litoria aurea) at the individual level based on tail venation patterns. We collected photographs of the tails of captive-raised tadpoles using a smartphone over a 4-week period. This photo-library was used to create an electronic survey where participants were asked to detect matches for query tadpoles from small image pools. We found that most participants agreed on a match for each query, with perfect consensus achieved for most queries (83%). We detected a 14% decline in perfect consensus when participants were asked to match images of tadpoles separated by longer time intervals, suggesting that it is more difficult to visually identify recapture events of L. aurea tadpoles over extended periods due to changes to tail appearance. However, consensus was obtained by participants for all queries, with all matches verified as being correct by the primary researcher. The strength of agreement among participants with no prior experience in matching tadpole tails suggests that there is sufficient inter-individual variation in this feature for individuals to be manually identified. We thus propose that photo-identification is likely to be a valid, non-invasive technique that can be used for short-term studies on tadpole populations that display tail venation. This offers an alternative to artificial markers that may not allow for individual identification, while also opening up tadpole monitoring programmes to citizen scientists who can be recruited online to process image data from home.     

The BAM H1 Restriction Site in the Bacteriophage T4 Chromosome Is Located in or near Gene 8

Bacteriophage T4 cytosine-containing DNA is cleaved at a single site by the restriction endonuclease, Bam H1. The site lies within the late region of the T4 genome, close to, or within, gene 8, one of the structural genes of the phage particle baseplate.     

Identifying disease-associated biomarker network features through conditional graphical model

Biomarkers are often organized into networks, in which the strengths of network connections vary across subjects depending on subject-specific covariates (eg, genetic variants). Variation of network connections, as subject-specific feature variables, has been found to predict disease clinical outcome. In this work, we develop a two-stage method to estimate biomarker networks that account for heterogeneity among subjects and evaluate network's association with disease clinical outcome. In the first stage, we propose a conditional Gaussian graphical model with mean and precision matrix depending on covariates to obtain covariate-dependent networks with connection strengths varying across subjects while assuming homogeneous network structure. In the second stage, we evaluate clinical utility of network measures (connection strengths) estimated from the first stage. The second-stage analysis provides the relative predictive power of between-region network measures on clinical impairment in the context of regional biomarkers and existing disease risk factors. We assess the performance of proposed method by extensive simulation studies and application to a Huntington's disease (HD) study to investigate the effect of HD causal gene on the rate of change in motor symptom through affecting brain subcortical and cortical gray matter atrophy connections. We show that cortical network connections and subcortical volumes, but not subcortical connections are identified to be predictive of clinical motor function deterioration. We validate these findings in an independent HD study. Lastly, highly similar patterns seen in the gray matter connections and a previous white matter connectivity study suggest a shared biological mechanism for HD and support the hypothesis that white matter loss is a direct result of neuronal loss as opposed to the loss of myelin or dysmyelination.     

Economic Evaluation of a Community‐based Obesity Prevention Program in Children: The APPLE Project

Effective strategies are urgently required to reduce the prevalence of obesity during growth. Determining which strategies are most successful should also include analysis of their relative costs. To date, few obesity prevention studies in children have reported data concerning cost‐effectiveness. The aim of this study was to assess the costs and health benefits of implementing the APPLE (A Pilot Program for Lifestyle and Exercise) project, a 2‐year controlled community‐based obesity prevention initiative utilizing activity coordinators (ACs) in schools and nutrition promotion in New Zealand children (5–12 years). The marginal costs of the project in 2006 prices were estimated and compared with the kilograms (kg) of weight‐gain prevented for children in the intervention relative to the control arm. The children's health‐related quality of life (HRQoL) was also measured using the Health Utilities Index (HUI). The total project cost was NZ$357,490, or NZ$1,281 per intervention child for 2 years (NZ$1 = US$0.67 = UK£0.35 = EUR €0.52). Weight z‐score was reduced by 0.18 (0.13, 0.22) units at 2 years and 0.17 (0.11, 0.23) units at 4 years in intervention relative to control children. Mean HUI values did not differ between intervention and control participants. The reduction in weight z‐score observed is equivalent to 2.0 kg of weight‐gain prevented at 15 years of age. The relatively simple intervention approach employed by the APPLE project was successful in significantly reducing the rate of excessive weight gain in children, with implementation costs of NZ$664–1,708 per kg of weight‐gain prevented over 4 years.     

Milk-Derived Antimicrobial Peptides: Overview,Applications, and Future Perspectives

Probiotics and Antimicrobial Proteins - The growing consumer awareness towards healthy and safe food has reformed food processing strategies. Nowadays, food processors are aiming at natural,...