Expression and purification of functional human glycogen synthase-1:glycogenin-1 complex in insect cells

We report the successful expression and purification of functional human muscle glycogen synthase (GYS1) in complex with human glycogenin-1 (GN1). Stoichiometric GYS1:GN1 complex was produced by co-expression of GYS1 and GN1 using a bicistronic pFastBac™-Dual expression vector, followed by affinity purification and subsequent size-exclusion chromatography. Mass spectrometry analysis identified that GYS1 is phosphorylated at several well-characterised and uncharacterised Ser/Thr residues. Biochemical analysis, including activity ratio (in the absence relative to that in the presence of glucose-6-phosphate) measurement, covalently attached phosphate estimation as well as phosphatase treatment, revealed that recombinant GYS1 is substantially more heavily phosphorylated than would be observed in intact human or rodent muscle tissues. A large quantity of highly-pure stoichiometric GYS1:GN1 complex will be useful to study its structural and biochemical properties in the future, which would reveal mechanistic insights into its functional role in glycogen biosynthesis

Isomorphic factorisations V: Directed graphs

An isomorphic factorisation of a digraph D is a partition of its arcs into mutually isomorphic subgraphs. If such a factorisation of D into exactly t parts exists, then t must divide the number of arcs in D. This is called the divisibility condition. It is shown conversely that the divisibility condition ensures the existence of an isomorphic factorisation into t parts in the case of any complete digraph. The sufficiency of the divisibility condition is also investigated for complete m‐partite digraphs. It is shown to suffice when m = 2 and t is odd, but counterexamples are provided when m = 2 and t is even, and when m = 3 and either t = 2 or t is odd.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/152659/1/mtks0025579300009529.pd

Effects of wheat and oat-based whole grain foods on serum lipoprotein size and distribution in overweight middle aged people : a randomised controlled trial

Peer reviewedPublisher PD

Linkage disequilibrium on chromosome 6 in Australian Holstein-Friesian cattle

We analysed linkage disequilibrium (LD) in Australian Holstein-Friesian cattle by genotyping a sample of 45 bulls for 15 closely-spaced microsatellites on two regions of BTA6 reported to carry important QTL for dairy traits. The order and distance of markers were based on the USDA-MARC linkage map. Frequencies of haplotypes were estimated using the E-M approach and a more computationally-intensive Bayesian approach as implemented in PHASE. LD was then estimated using the Hedrick multiallelic extension of Lewontin normalised coefficient D'. Estimates of D' from the two approaches were in close agreement (r = 0.91). The mean estimates of D' for marker pairs with an inter-marker distance of less than 5 cM (n = 13) are 0.57 and 0.51, and for distances more than 20 cM (n = 44) are 0.29 and 0.17, estimated from the E-M and Bayesian approaches, respectively. The Malecot model was fitted for the exponential decline of LD with map distance between markers. The swept radii (the distance at which LD has declined to 1/e (~37%) of its initial value) are 11.6 and 13.7 cM for the above two methods, respectively. The Malecot model was also fitted using map distance in Mb from the bovine integrated map (bovine location database, bLDB) in addition to cM from the MARC map. Overall, the results indicate a high level of LD on chromosome 6 in Australian dairy cattle

Is the Sevier Desert Reflection of West-Central Utah a Normal Fault?: Comment and Reply

Forum discussion on an article originally by Anders et al. Allmendinger and Royse critique Anders et al.'s argument, to which Anders et al. responds. Critique abstract: The continuing discussion of the Sevier Desert region, almost 20 years after MacDonald’s (1976) classic paper, provides a measure of the significance of the region as well as the non-uniqueness of seismic reflection data interpretation. The article by Anders and Christie-Blick (1994) and the nearly simultaneous publication of similar ideas by Hamilton (1994) raise important questions. The interpretation of a Sevier Desert detachment has always been a hypothesis to be tested rather than a fact. In our opinion, however, Anders and Christie-Blick and Hamilton have ignored a variety of basic geologic data requiring the presence of a major low-angle normal fault on the east side of the Sevier Desert basin. Response abstract: We concur with Allmendinger and Royse’s assessment of the detachment interpretation for the Sevier Desert reflection as a ‘‘hypothesis to be tested.’’ We reported on an attempt to do just that: to look for evidence for fault-related deformation in samples from two boreholes that intersect this feature. The absence of evidence for cataclasis in the inferred hanging-wall block or ductile deformation in the footwall naturally raises some interesting issues for the tectonic interpretation of the Sevier Desert. In drawing attention to these issues, we have not ‘‘ignored’’ any basic geologic data, nor are we aware of any data that ‘‘require’’ the presence of a major low-angle normal fault

Pituitary Lesions, Obesity, and Mesenteric Lipomas in Insulin-Resistant Horses

The aim of the current study was to identify associations between pituitary lesions, body condition scores, and mesenteric lipomas in horses with insulin resistance. Necropsy examinations were performed following euthanasia in 30 adult horses designated as insulin resistant (n = 11) or insulin sensitive (n = 19). Insulin sensitivity was determined using the insulin-modified frequently sampled intravenous glucose tolerance test and resting insulin concentrations. At necropsy, mesenteric lipomas were measured. The pituitary and adrenal glands, pancreas, and liver were evaluated histologically; pituitary glands were scored based on published criteria. Insulin-resistant horses had significantly higher pituitary scores (p = 0.0035) and body condition scores (p = 0.0001), even when adjusting for age, and a greater frequency of mesenteric lipomas (p = 0.014) and greater lipoma area (p = 0.0332) than insulin-sensitive horses. Regardless of insulin status, horses with pituitary scores ≥3 (diffuse hyperplasia; n = 25) had higher body condition scores (p = 0.0313) and a greater frequency of mesenteric lipomas (p \u3c 0.0002) than those with lower pituitary scores. High body condition score was not correlated to an increased frequency of mesenteric lipomas. Detection of higher pituitary scores in insulin-resistant horses suggested an association between insulin resistance and pituitary morphology. Horses in the insulin-resistant group and those with high pituitary scores had higher body condition scores and a greater frequency of mesenteric lipomas. These horses might be at increased risk for lipoma-associated colic

A Novel Vertebrate Eye Using Both Refractive and Reflective Optics

SummarySunlight is attenuated rapidly in the ocean, resulting in little visually useful light reaching deeper than ∼1000 m in even the clearest water [1]. To maximize sensitivity to the relatively brighter downwelling sunlight, to view the silhouette of animals above them, and to increase the binocular overlap of their eyes, many mesopelagic animals have developed upward-pointing tubular eyes [2–4]. However, these sacrifice the ability to detect bioluminescent [5] and reflective objects in other directions. Thus, some mesopelagic fish with tubular eyes extend their visual fields laterally and/or ventrally by lensless ocular diverticula, which are thought to provide unfocused images, allowing only simple detection of objects, with little spatial resolution [2–4]. Here, we show that a medial mirror within the ventrally facing ocular diverticulum of the spookfish, Dolichopteryx longipes, consisting of a multilayer stack derived from a retinal tapetum, is used to reflect light onto a lateral retina. The reflective plates are not orientated parallel to the surface of the mirror. Instead, plate angles change progressively around the mirror, and computer modeling indicates that this provides a well-focused image. This is the first report of an ocular image being formed in a vertebrate eye by a mirror

NIHTS: the near-infrared high throughput spectrograph for the Discovery Channel Telescope

NIHTS is a first-generation instrument now in use on Lowell Observatory’s Discovery Channel Telescope. It is a nearinfrared prism spectrograph of the BASS design featuring high throughput and low dispersion that is intended for observations of faint solar system and astrophysical objects over the YJHK spectral range. An unusual feature is its ability to observe simultaneously with the Large Monolithic Imager, an optical CCD camera, by means of a dichroic fold mirror. This is particularly valuable for time-variable targets such as Kuiper Belt Objects, asteroids, exoplanet transits, and brown dwarfs. We describe its design details and performance both in the lab and on the telescope

Hypercholesterolemia and chronic ischemia alter myocardial responses to selective cyclooxygenase-2 inhibition

ObjectiveCyclooxygenase-2 inhibitors have been implicated in adverse cardiac events. We hypothesize that hypercholesterolemia and ischemia may alter the myocardial response to the cyclooxygenase-2 inhibitor celecoxib.MethodsYorkshire swine fed normal chow (CX, n = 6) or high-cholesterol diet (HCX, n = 6) underwent placement of an Ameroid constrictor on the left circumflex artery and were started on celecoxib (200 mg/day). After 7 weeks, ischemic and nonischemic myocardium was analyzed for thrombogenic ratio (thromboxane content divided by prostacyclin content), total protein oxidative stress, and expression of prostacyclin synthase, thromboxane synthase, myeloperoxidase, and superoxide dismutase. Cardiac function, tissue perfusion, and vessel density were measured.ResultsHCX animals were significantly hypercholesterolemic compared with CX animals. Thrombogenic ratio was significantly higher in the HCX group than in the CX group, but prostacyclin and thromboxane synthase expression was similar in all tissues. Myocardial perfusion was decreased in the HCX group compared with the CX group. Total oxidative stress, myeloperoxidase, and superoxide dismutase were increased in ischemic tissue compared with nonischemic tissues, but there was no diet-induced difference between groups. There was no difference in capillary or arteriolar density between groups. Left ventricular contractility was greater in the HCX group than in the CX group, but there was no significant difference in heart rate, mean arterial pressure, or left ventricular pressure.ConclusionsHypercholesterolemic patients using celecoxib may be at higher risk for thrombotic events than those with normal cholesterol, but the relationship between dyslipidemia, ischemia, and cyclooxygenase-2 inhibition is likely much more complicated than originally thought