Mycobacterial Ku and ligase proteins constitute a two-component NHEJ repair machine

In mammalian cells, repair of DNA double-strand breaks (DSBs) by nonhomologous end-joining (NHEJ) is critical for genome stability. Although the end-bridging and ligation steps of NHEJ have been reconstituted in vitro, little is known about the end-processing reactions that occur before ligation. Recently, functionally homologous end-bridging and ligation activities have been identified in prokarya. Consistent with its homology to polymerases and nucleases, we demonstrate that DNA ligase D from Mycobacterium tuberculosis (Mt-Lig) possesses a unique variety of nucleotidyl transferase activities, including gap-filling polymerase, terminal transferase, and primase, and is also a 3' to 5' exonuclease. These enzyme activities allow the Mt-Ku and Mt-Lig proteins to join incompatible DSB ends in vitro, as well as to reconstitute NHEJ in vivo in yeast. These results demonstrate that prokaryotic Ku and ligase form a bona fide NHEJ system that encodes all the recognition, processing, and ligation activities required for DSB repair.     

Continuous lumbar hemilaminectomy for intervertebral disc disease in an Amur tiger (Panthera tigris altaica)

A 13-yr-old Amur tiger (Panthera tigris altaica) was presented for an acute onset of paraplegia. Spinal imaging that included plain radiographs, myelography, and computed tomography performed under general anesthesia revealed lateralized spinal cord compression at the intervertebral disc space L4-5 caused by intervertebral disc extrusion. This extrusion was accompanied by an extensive epidural hemorrhage from L3 to L6. Therefore, a continuous hemilaminectomy from L3 to L6 was performed, resulting in complete decompression of the spinal cord. The tiger was ambulatory again 10 days after the surgery. This case suggests that the potential benefit of complete spinal cord decompression may outweigh the risk of causing clinically significant spinal instability after extensive decompression.     

Regional and local scale variations in soil organic carbon stocks in West Greenland

The soil organic carbon (SOC) pool of the Northern Hemisphere contains about half of the global SOC stored in soils. As the Arctic is exceptionally sensitive to global warming, temperature rise and prolonged summer lead to deeper thawing of permafrost‐affected soils and might contribute to increasing greenhouse gas emissions progressively. To assess the overall feedback of soil organic carbon stocks (SOCS) to global warming in permafrost‐affected regions the spatial variation in SOCS at different environmental scales is of great interest. However, sparse and unequally distributed soil data sets at various scales in such regions result in highly uncertain estimations of SOCS of the Northern Hemisphere and here particularly in Greenland. The objectives of this study are to compare and evaluate three controlling factors for SOCS distribution (vegetation, landscape, aspect) at two different scales (local, regional). The regional scale reflects the different environmental conditions between the two study areas at the coast and the ice margin. On the local scale, characteristics of each controlling factor in form of defined units (vegetation units, landscape units, aspect units) are used to describe the variation in the SOCS over short distances within each study area, where the variation in SOCS is high. On a regional scale, we investigate the variation in SOCS by comparing the same units between the study areas. The results show for both study areas that SOCS are with 8 kg m^?2 in the uppermost 25 cm and 16 kg m^?2 in the first 100 cm of the soil, i.e., 3 to 6 kg m^?2 (37.5%) higher than existing large scale estimations of SOCS in West Greenland. Our approach allows to rank the scale‐dependent importance of the controlling factors within and between the study areas. However, vegetation and aspect better explain variations in SOCS than landscape units. Therefore, we recommend vegetation and aspect for determining the variation in SOCS in West Greenland on both scales.     

ULTRASONOGRAPHIC EXAMINATION OF THE THYROID GLAND OF HYPERTHYROID CATS: COMPARISON TO 99mTcO−4 SCINTIGRAPHY

High-resolution ultrasonography was evaluated as an alternative to ^99mTcO^-₄ scintigraphy for examining size and appearance of thyroid glands in hyperthyroid cats. Thyroid ultrasound examinations were performed on 6 normal cats and 14 cats with hyperthyroidism. Thyroid lobe volume was estimated from ultrasound images using the equation for a prolate ellipsoid, π/6 (length * height * width). Total thyroid volume was estimated by adding the volume estimations of the left and right lobes. Thyroid lobes of hyperthyroid cats were considered abnormal if estimated volume exceeded the 99% confidence interval for normal thyroid volume determined from the control group. Scintigraphic examinations performed on hyperthyroid cats were evaluated for unilateral versus bilateral disease and for the presence of ectopic activity. Mean thyroid lobe volume and total thyroid volume for normal cats was 85 and 169 mm³, respectively. Mean thyroid lobe volume and total thyroid volume for hyperthyroid cats was 578 and 889 mm³. There was a significant difference in mean estimated total thyroid volume of normal and hyperthyroid cats. Thyroid lobes with greater than normal TcO^-₄ uptake on scintigraphy were larger and had variable homogeneity, echogenicity, and margination on ultrasound examination. There also was an 85.7% agreement of scintigraphy and ultrasonography in differentiating normal from abnormal thyroid lobes. A fair correlation between estimated total thyroid volume of hyperthyroid cats and most recent pretherapy serum thyroxine values were also found. This preliminary study indicates that thyroid ultrasound examination may provide information that is useful for diagnosis and treatment of feline hyperthyroidism. Although ultrasound provides accurate evaluation of the thyroid glands, it cannot replace ^99mTcO^-₄ scintigraphy for screening of metastatic lesions and ectopic glands.     

Notification for intended animal research - remarks on the future procedures on the basis of the European Union Laboratory Animal Guideline 2010/63/EU

The new Directive 2010/63/EU on the protection of animals used for scientific purposes causes a need of several changes in German animal welfare law. On the basis of the regulations in the Directive, adjustments of the existing notification procedures are unavoidable. The Directive includes provisions for a simplified administrative procedure which differs significantly from the existing notification procedures. In any case, an application process will be required for projects in the meaning of the Directive. This contribution comments on the conceivable changes of the existing notification procedure. The implementation of the provisions of the Directive into national law has to be done near-term as the Directive is applicable from January 1st 2013. In advance, political and legal procedures must be passed through, the adapted regulations are to be notified by the European Commission and last but not least the authorities and user establishments must have enough time to implement the necessary changes.     

Dynamic visualization of thrombopoiesis within bone marrow

Platelets are generated from megakaryocytes (MKs) in mammalian bone marrow (BM) by mechanisms that remain poorly understood. Here we describe the use of multiphoton intravital microscopy in intact BM to visualize platelet generation in mice. MKs were observed as sessile cells that extended dynamic proplatelet-like protrusions into microvessels. These intravascular extensions appeared to be sheared from their transendothelial stems by flowing blood, resulting in the appearance of proplatelets in peripheral blood. In vitro, proplatelet production from differentiating MKs was enhanced by fluid shear. These results confirm the concept of proplatelet formation in vivo and are consistent with the possibility that blood flow-induced hydrodynamic shear stress is a biophysical determinant of thrombopoiesis.     

Capillary wrinkling of floating thin polymer films

A freely floating polymer film, tens of nanometers in thickness, wrinkles under the capillary force exerted by a drop of water placed on its surface. The wrinkling pattern is characterized by the number and length of the wrinkles. The dependence of the number of wrinkles on the elastic properties of the film and on the capillary force exerted by the drop confirms recent theoretical predictions on the selection of a pattern with a well-defined length scale in the wrinkling instability. We combined scaling relations that were developed for the length of the wrinkles with those for the number of wrinkles to construct a metrology for measuring the elasticity and thickness of ultrathin films that relies on no more than a dish of fluid and a low-magnification microscope. We validated this method on polymer films modified by plasticizer. The relaxation of the wrinkles affords a simple method to study the viscoelastic response of ultrathin films.     

Influence of cereal non-starch polysaccharides on ileo-caecal and rectal microbial populations in growing pigs

The effect of cereal non-starch polysaccharides (NSP) on the gut microbial populations was studied in 5 growing pigs between 39-116 kg body weight according to a Latin square design. The diets were composed to contain different NSP levels. The control diet had a normal NSP content (139 g/kg dry matter (DM)), 2 diets had a low total amount of NSP (95 and 107 g/kg DM) and 2 diets had a high amount of total NSP (191 and 199 g/kg DM). Furthermore, one of the diets within each category had a content of insoluble NSP similar to the control diet and one had a high content of insoluble NSP. Samples were collected from the ileum, via intestinal post valve T-caecum (PVTC) cannulas surgically inserted at the ileo-caecal ostium, and from the rectum. The total microbial flora of the ileal samples were analysed for by defining base pair length with terminal restriction fraction length polymorphism (T-RFLP). The microbial diversity of the coliform flora of the ileal and rectal samples were defined by biochemical fingerprinting. It was observed that many terminal restriction fragments (TRFs) disappeared when new diets were introduced and that some characteristic TRFs were found in the high and low NSP diets, respectively. Both the total gut microflora and the coliform flora were influenced by the dietary NSP content.     

Entropically stabilized local dipole formation in lead chalcogenides

We report the observation of local structural dipoles that emerge from an undistorted ground state on warming, in contrast to conventional structural phase transitions in which distortions emerge on cooling. Using experimental and theoretical probes of the local structure, we demonstrate this behavior in binary lead chalcogenides, which were believed to adopt the ideal, undistorted rock-salt structure at all temperatures. The behavior is consistent with a simple thermodynamic model in which the emerging dipoles are stabilized in the disordered state at high temperature due to the extra configurational entropy despite the fact that the undistorted structure has lower internal energy. Our findings shed light on the anomalous electronic and thermoelectric properties of the lead chalcogenides. Similar searches may show that the phenomenon is more widespread.