Assessment of bone healing during callus distraction by an automatic torsional stiffness metering system]

The present article describes a newly developed device for the quantitative assessment of torsional in vivo stiffness of regenerating bone under callus distraction. Both the design and function of this device, and its use during bony consolidation are discussed. The device exhibited an accuracy of +/- 18% for stiffness under 0.1 Nm/degree, and +/- 5% stiffness above 0.1 Nm/degree. The average accuracy was +/- 14%. The data scatter for the stiffness measurement ranged between +/- 1.43% and +/- 7.68% (average: +/- 3.99%). The precision of a test machine was between +/- 0.01% and +/- 11.3% (average: +/- 3.65%). The method has the following advantages over existing methods for investigating healing: 1. no need to dismantle the external fixation for measurement; 2. preservation of the bone axis with minimal risk of misalignment during the bone healing process; 3. minimal technical requirements, with easy, noninvasive measurement; 4. no exposure to X-radiation.     

Isolation and partial characterization of a Taenia taeniaeformis metacestode proteinase inhibitor

Suquet C., Green-Edwards C. and Wes Leid R. 1984. Isolation and partial characterization of a Taenia taeniaeformis metacestode proteinase inhibitor. International Journal for Parasitology14: 165–172. A proteinase inhibitor from the metacestode of Taenia taeniaeformis was purified 136-fold to apparent homogeneity as evidenced by one Coomassie Blue protein staining band on 10% SDS slab gels under both reducing and non-reducing conditions. The apparent molecular weight under dissociating conditions was 19,500. This parasite protein inhibited esterolysis of TAME and BTEE by bovine pancreatic trypsin and chymotrypsin respectively in a time and dose-dependent manner. Proteolysis of casein by both enzymes was also inhibited in a time and dose-dependent manner. The parasite inhibitor was stable from pH 2.2 to 10.5 and was fully active after heating at 56 °C for 3 h. The proteases pronase and thermolysin, at concentrations of 1 mg ml^?1, completely inactivated the metacestode inhibitor. Two sulfhydryl proteases, papain and chymopapain, used at concentrations of 1 mg ml^?1 were without effect. The irreversible proteinase inhibitors TLCK, TPCK and PMSF at concentrations up to 10 mM had no effect on the parasite inhibitor.     

Using comparative genomics to reorder the human genome sequence into a virtual sheep genome

Background

Is it possible to construct an accurate and detailed subgene-level map of a genome using bacterial artificial chromosome (BAC) end sequences, a sparse marker map, and the sequences of other genomes?

Results

A sheep BAC library, CHORI-243, was constructed and the BAC end sequences were determined and mapped with high sensitivity and low specificity onto the frameworks of the human, dog, and cow genomes. To maximize genome coverage, the coordinates of all BAC end sequence hits to the cow and dog genomes were also converted to the equivalent human genome coordinates. The 84,624 sheep BACs (about 5.4-fold genome coverage) with paired ends in the correct orientation (tail-to-tail) and spacing, combined with information from sheep BAC comparative genome contigs (CGCs) built separately on the dog and cow genomes, were used to construct 1,172 sheep BAC-CGCs, covering 91.2% of the human genome. Clustered non-tail-to-tail and outsize BACs located close to the ends of many BAC-CGCs linked BAC-CGCs covering about 70% of the genome to at least one other BAC-CGC on the same chromosome. Using the BAC-CGCs, the intrachromosomal and interchromosomal BAC-CGC linkage information, human/cow and vertebrate synteny, and the sheep marker map, a virtual sheep genome was constructed. To identify BACs potentially located in gaps between BAC-CGCs, an additional set of 55,668 sheep BACs were positioned on the sheep genome with lower confidence. A coordinate conversion process allowed us to transfer human genes and other genome features to the virtual sheep genome to display on a sheep genome browser.

Conclusion

We demonstrate that limited sequencing of BACs combined with positioning on a well assembled genome and integrating locations from other less well assembled genomes can yield extensive, detailed subgene-level maps of mammalian genomes, for which genomic resources are currently limited.     

Expansion of effector memory regulatory T cells represents a novel prognostic factor in lower risk myelodysplastic syndrome

Myelodysplastic syndromes are premalignant diseases characterized by cytopenias, myeloid dysplasia, immune dysregulation with association to autoimmunity, and variable risk for acute myeloid leukemia transformation. Studies of FOXP3(+) regulatory T cells (Tregs) indicate that the number and/or activation state may influence cancer progression in these patients. Focusing on patients with a lower risk for leukemia transformation, 18 (34.6%) of 52 patients studied displayed an altered Treg compartment compared with age-matched controls. Delineation of unique Treg subsets revealed that an increase in the absolute number of CD4(+)FOXP3(+)CD25(+)CD127(low)CD45RA(-)CD27(-) Tregs (effector memory Tregs [Treg(EM)]) was significantly associated with anemia (p = 0.046), reduced hemoglobin (p = 0.038), and blast counts ≥5% (p = 0.006). In healthy donors, this Treg(EM) population constitutes only 2% of all Tregs (one to six Tregs per microliter) in peripheral blood but, when isolated, exhibit greater suppressive activity in vitro. With a median follow-up of 3.1 y (range 2.7-4.9 y) from sample acquisition, increased numbers of Treg(EM) cells proved to have independent prognostic importance in survival estimates, suggesting that enumeration of this Treg subset may be a more reliable indicator of immunological escape than FOXP3(+) T cells as a whole. Based on multivariate analyses, Treg(EM) impacted survival independently from myeloblast characteristics, cytopenias, karyotype, and comorbidities. Based on these findings, Treg(EM) cell expansion may be synonymous with human Treg activation and indicate microenvironmental changes conducive to transformation in myelodysplastic syndromes.     

The Visible,Sustainable Farm: A Comprehensive Energy Analysis of a Midwestern Farm

The Sunshine Farm, in central Kansas, offers a unique data set detailing all the inputs and outputs of a farming project intended to be based, as much as possible, on solar energy. Such a complete and detailed data set, encompassing more than 1.25 M data points for an 85-ha, 7-year project, has never before been compiled for any farm. The data show that important energy inputs that have thus far been left out of existing farm energy efficiency estimates, with implications for biofuels and other biomass for energy technologies. The SSF achieved energy efficiencies superior to conventional agriculture while maintaining soil health and delivering more nutritious, organic products. A first-ever analysis of the energy efficiency of horse traction shows that horses are significantly less energy efficient and much less labor efficient than tractors, although the horses in the study were clearly underutilized. An analysis of the farm removing all inputs and outputs relevant to the horses shows that without horses the Sunshine Farm would have been competitive with conventional energy efficiencies even taking into account the farm's higher labor inputs and small size.     

Wavelet analysis of scaling properties of gastric electrical activity.

We present a novel approach to the analysis of fluctuations in human myoelectrical gastric activity measured noninvasively from the surface of the abdomen. The time intervals between successive maxima of the wavelet transformed quasi-periodic electrogastrographic waveform define the gastric rate variability (GRV) time series. By using the method of average wavelet coefficients, the statistical fluctuations in the GRV signal in healthy individuals are determined to scale in time. Such scaling was previously found in a variety of physiological phenomena, all of which support the hypothesis that physiological dynamics utilize fractal time series. We determine the scaling index in a cohort of 17 healthy individuals to be 0.80 +/- 0.14, which compared with a set of surrogate data is found to be significant at the level P < 0.01. We also determined that the dynamical pattern, so evident in the spectrum of average wavelet coefficients of the GRV time series of healthy individuals, is significantly reduced in a cohort of systemic sclerosis patients having a scaling index 0.64 +/- 0.17. These results imply that the long-term memory in GRV time series is significantly reduced from healthy individuals to those with systemic sclerosis. Consequently, this disease degrades the complexity of the underlying gastrointestinal control system and this degradation is manifest in the loss of scaling in the GRV time series.     

Minimotif Miner: a tool for investigating protein function

In addition to large domains, many short motifs mediate functional post-translational modification of proteins as well as protein-protein interactions and protein trafficking functions. We have constructed a motif database comprising 312 unique motifs and a web-based tool for identifying motifs in proteins. Functional motifs predicted by MnM can be ranked by several approaches, and we validated these scores by analyzing thousands of confirmed examples and by confirming prediction of previously unidentified 14-3-3 motifs in EFF-1.