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Fifteen rumen fistulated Holstein cows in late lactation and fed a total mixed ration offered ad libitum were supplemented with Se yeast to provide 0, 11, 20, 30, or 42 mg of supplemental Se/day to test the hypothesis that amounts of Se secreted in milk, excreted in urine and feces, and apparently retained in tissues would increase in direct proportion to Se intake. One-half of the yeast supplement was placed directly into the rumen through the fistula of each cow just before milking in the morning and again in the evening, and estimates of average daily excretion of Se were made using total collections of urine and feces from 25 to 31 d after treatments commenced. Amounts of Se secreted daily in milk and apparently retained in tissues increased linearly with average daily intake of Se. The amount of Se excreted in feces and total excretion of Se in urine plus feces increased curvilinearly with Se intake, such that proportionately less Se was excreted as the amount of Se fed increased. On average, total Se excretion accounted for 66%, Se secretion in milk accounted for 17%, and Se apparently retained in tissues accounted for 17% of total Se intake by cows. Thus, in herds fed large amounts of Se yeast, most of the Se will be excreted and retained on-farm. High concentrations of Se will be found where urine and feces accumulate (e.g., yards and effluent ponds), and effluent management practices must be tailored to avoid environmental issues., (Copyright © 2010 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Nuclear DBF-2-related (NDR) kinases are essential regulators of cell cycle progression, growth, and development in many organisms and are activated by the binding of an Mps One Binder (MOB) protein partner, autophosphorylation, and phosphorylation by an upstream STE20 family kinase. In the protozoan parasite, Trypanosoma brucei, the causative agent of human African trypanosomiasis, the NDR kinase, PK50, is expressed in proliferative life cycle stages and was shown to complement a yeast NDR kinase mutant cell line. However, the function of PK50 and a second NDR kinase, PK53, in T. brucei has not been determined to date, although trypanosome MOB1 is known to be essential for cytokinesis, suggesting the NDR kinases may also be involved in this process. Here, we show that specific depletion of PK50 or PK53 from bloodstream stage trypanosomes resulted in the rapid accumulation of cells with two nuclei and two kinetoplasts, indicating that cytokinesis was specifically inhibited. This led to a deregulation of the cell cycle and cell death and provides genetic validation of these kinases as potential novel drug targets for human African trypanosomiasis. Recombinant active PK50 and PK53 were produced and biochemically characterized. Both enzymes autophosphorylated, were able to trans-phosphorylate generic kinase substrates in vitro, and were active in the absence of phosphorylation by an upstream kinase. Additionally, both enzymes were active in the absence of MOB1 binding, which was also demonstrated to likely be a feature of the kinases in vivo. Biochemical characterization of recombinant PK50 and PK53 has revealed key kinetic differences between them, and the identification of in vitro peptide substrates in this study paves the way for high throughput inhibitor screening of these kinases.

The neural mechanisms underlying the integration and segregation of motion signals are often studied using plaid stimuli. These stimuli consist of two spatially coincident dynamic gratings of differing orientations, which are either perceived to move in two unique directions or are integrated by the visual system to elicit the percept of a checkerboard moving in a single direction. Computations pertaining to the motion of the individual component gratings are thought to take place in striate cortex (V1) whereas motion integration is thought to involve neurons in dorsal stream extrastriate visual areas, particularly V5/MT. By combining a psychophysical task that employed plaid stimuli with 1 Hz offline repetitive transcranial magnetic stimulation (rTMS), we demonstrated a double dissociation between striate and extrastriate visual cortex in terms of their contributions to motion integration. rTMS over striate cortex increased coherent motion percepts whereas rTMS over extrastriate cortex had the opposite effect. These effects were robust directly after the stimulation administration and gradually returned to baseline within 15 minutes. This double dissociation is consistent with previous patient data and the recent hypothesis that both coherent and transparent motion percepts are supported by the visual system simultaneously and compete for perceptual dominance.

Humans with amblyopia display anomalous performance for global motion discrimination. Attempts have been made to rule out an explanation based solely on the visibility loss in lower visual areas. However, it remains a possibility that the altered scale over which local motion is processed in V1 might lead to reduced efficiency of global motion processing in extra-striate cortex. We use stimuli composed of spatial frequency bandpass elements, equated for visibility, to show that the global motion deficit in amblyopia for both fellow and amblyopic eyes is still present once impairments in low-level processing have been factored out. This residual deficit appears to be spatial scale invariant and the relative deficit between the eyes shows a dependence on stimulus speed. We believe that this rules out an explanation of the amblyopic global motion deficit based solely on local motion input. We suggest instead that, in addition to low-level deficits, motion processing in a broadband, extra-striate, global motion mechanism is impaired in amblyopia.

Here we examine how global translational motion sensitivity varies with the spatial frequency of the elements in local motion and on the eccentricity of stimulation. Using DC-balanced, spatially narrowband elements (radial log Gabors) matched in terms of multiples above contrast threshold, we show that global translational motion sensitivity is best at mid high spatial frequencies and worst at low spatial frequencies. Furthermore, we show that the lower global motion sensitivity of the periphery is due to differences in spatial scale/contrast that can be attributed to lower reaches of the visual pathway where the local motion signal is transduced. Thus, the efficiency of the global translational motion computation that occurs in extrastriate cortical areas does not vary across the visual field. This may not be directly applicable to global radial motion because there are known visual field anisotropies.

Persian scholar Ibn al-Haytham ('Alhazen') has rightly been credited with many advances in optics and vision science, but recent spurious claims that he is the 'founder of psychophysics' rest upon unsupported assertions, a conflation of psychophysics with the wider discipline of psychology, and semantic arguments over what it is to 'found' a school of thought.

Purpose: Amblyopic observers show deficits for global motion discrimination that cannot be accounted for by their contrast sensitivity impairment. The processing of first- and second-order translational global motion is deficient, as is the processing of first-order optic flow, suggesting that cortical function in extrastriate areas is impaired. The authors sought to determine whether amblyopes show impairment in the processing of optic flow defined by second-order motion, whether these deficits are comparable in the two eyes, and whether these deficits are correlated with first-order deficits., Methods: Eight amblyopic subjects (three strabismic, three strabismic-anisometropic, one anisometropic, one deprivation; mean age, 29 years) were tested. The authors used random dot kinematograms in which the dots were luminance or contrast modulations of background noise. The global pattern of dot motion within the stimulus area was translational, radial, or rotational. Coherence thresholds for direction discrimination were obtained across a range of dot modulation depths, allowing the separation of contrast and motion deficits., Results: The present study showed that deficits in second-order optic flow processing were equivalent to those for first-order stimuli and that these were unrelated to the extent of the amblyopic contrast sensitivity deficit and were comparable in both eyes. Radial optic flow was more affected than rotational optic flow., Conclusions: Global motion impairment appeared to have a high-level binocular locus and was independent of the depth of the contrast deficit. Results also support the idea that global motion and optic flow processing are form-cue invariant.

Optic flow-large-field rotational and radial motion-is processed as efficiently as translational motion for first-order (luminance-defined) stimuli. However, it has been suggested recently that the same pattern does not hold for second-order (e.g. contrast-defined) stimuli. We used random dot kinematogram (RDK) stimuli to determine whether global processing of optic flow is as efficient as processing of global translational motion for both first- and second-order stimuli. For first-order stimuli, we found that coherence thresholds for radial and rotational motion were equivalent to thresholds for translational motion, supporting previous findings. For second-order stimuli we found, firstly, that given sufficient contrast, second-order optic flow can be processed as efficiently as first-order optic flow and, secondly, that rotational and translational second-order motion are processed with equal efficiency. This contradicts the suggestion that there is a loss of efficiency between integration of second-order global motion and second-order optic flow. The third interesting finding was that the processing of radial second-order motion appears to suffer from a deficit that is dependent upon both the contrast and spatial extent of the stimulus. Further experiments discounted the possibility that the observed deficit is caused by a centrifugal or centripetal bias, but demonstrated that a longer temporal integration period for radial second-order motion is responsible for the observed difference. For durations of approximately 850ms, all three types of motion are processed with equal efficiency.

ISWI proteins form the catalytic core of a subset of ATP-dependent chromatin remodeling activities in eukaryotes from yeast to man. Many of these complexes have been found to reposition nucleosomes but with different directionalities. We find that the yeast Isw1a, Isw2, and Chd1 enzymes preferentially move nucleosomes toward more central locations on short DNA fragments whereas Isw1b does not. Importantly, the inherent positioning properties of the DNA play an important role in determining where nucleosomes are relocated to by all of these enzymes. However, a key difference is that the Isw1a, Isw2, and Chd1 enzymes are unable to move nucleosomes to positions closer than 15 bp from a DNA end, whereas Isw1b can. We also find that there is a correlation between the inability of enzymes to move nucleosomes close to DNA ends and the preferential binding to nucleosomes bearing linker DNA. These observations suggest that the accessibility of linker DNA together with the positioning properties of the underlying DNA play important roles in determining the outcome of remodeling by these enzymes.

The two-stage decomposition-recombination model of 2D motion perception has been criticised on the basis that the direction of plaid stimuli can be accurately discriminated at speeds so low that the direction of their Fourier components is not discriminable. The nature of this gap in performance between gratings and plaids was investigated across a range of spatial frequencies and durations for first- and second-order stimuli. Motion-detection thresholds were obtained using a 2AFC, constant stimuli procedure and it was found that although thresholds for detection of plaid motion were often lower than those for gratings, the gap in performance between first-order plaids and gratings was unreliable, varying in magnitude and occasionally direction with the spatial frequency of the stimulus, presentation duration and observer. Curiously, an analogous gap found between purely second-order gratings and second-order plaids was more reliable and stable. It has been suggested that the gap is the result of 'local motion detectors' or broadly tuned V1 cells. The data presented here suggest that second-order mechanisms are responsible for the gap and that first-order information may even disrupt it.

The Snf2 family represents a functionally diverse class of ATPase sharing the ability to modify DNA structure. Here, we use a magnetic trap and an atomic force microscope to monitor the activity of a member of this class: the RSC complex. This enzyme caused transient shortenings in DNA length involving translocation of typically 400 bp within 2 s, resulting in the formation of a loop whose size depended on both the force applied to the DNA and the ATP concentration. The majority of loops then decrease in size within a time similar to that with which they are formed, suggesting that the motor has the ability to reverse its direction. Loop formation was also associated with the generation of negative DNA supercoils. These observations support the idea that the ATPase motors of the Snf2 family of proteins act as DNA translocases specialized to generate transient distortions in DNA structure.

In the 30 years since the discovery of the nucleosome, our picture of it has come into sharp focus. The recent high-resolution structures have provided a wealth of insight into the function of the nucleosome, but they are inherently static. Our current knowledge of how nucleosomes can be reconfigured dynamically is at a much earlier stage. Here, recent advances in the understanding of chromatin structure and dynamics are highlighted. The ways in which different modes of nucleosome reconfiguration are likely to influence each other are discussed, and some of the factors likely to regulate the dynamic properties of nucleosomes are considered.

During the conduct of an experiment designed to examine the nutritional management of dairy cows in late pregnancy, four cows out of 72 suffered from acute haemoglobinuria two to four weeks after calving. Thirty-six thin and 36 fat cows were individually fed one of three diets based on a total mixed ration with different energy or protein concentrations during the last 3 to 4 weeks before expected calving date. After calving, cows grazed pasture and were offered 6 kg dry matter of pelleted concentrates daily. The P concentrations of the feeds offered suggested that the cows' diets were marginally deficient in P relative to requirements. Plasma P concentrations were significantly (P < 0.05) lower in fat cows than in thin cows during the first 6 weeks of lactation (0.87 versus 1.12 mmol/L), but precalving diet had no effect (P > 0.05). Concentrations of plasma inorganic P of the four fat cows that developed acute haemoglobinuria were less than 0.3 mmol/L. However, plasma P concentrations in another 12 cows, none of which displayed overt symptoms, declined to similar levels. It appeared that inadequate dietary P may have predisposed cows to acute haemoglobinuria, but the precipitating cause was not readily obvious.

ATP-dependent chromatin remodeling activities function to manipulate chromatin structure during gene regulation. One of the ways in which they do this is by altering the positions of nucleosomes along DNA. Here we provide support for the ability of these complexes to move nucleosomes into positions in which DNA is unraveled from one edge. This is expected to result in the loss of histone-DNA contacts that are important for retention of one H2A/H2B dimer within the nucleosome. Consistent with this we find that several chromatin remodeling complexes are capable of catalyzing the exchange of H2A/H2B dimers between chromatin fragments in an ATP-dependent reaction. This provides eukaryotes with additional means by which they may manipulate chromatin structure.

The ISWI proteins form the catalytic core of a subset of ATP-dependent chromatin-remodeling activities. Here, we studied the interaction of the ISWI protein with nucleosomal substrates. We found that the ability of nucleic acids to bind and stimulate the ATPase activity of ISWI depends on length. We also found that ISWI is able to displace triplex-forming oligonucleotides efficiently when they are introduced at sites close to a nucleosome but successively less efficiently 30 to 60 bp from its edge. The ability of ISWI to direct triplex displacement was specifically impeded by the introduction of 5- or 10-bp gaps in the 3'-5' strand between the triplex and the nucleosome. In combination, these observations suggest that ISWI is a 3'-5'-strand-specific, ATP-dependent DNA translocase that may be capable of forcing DNA over the surface of nucleosomes.