Bayesian estimation of sensitivity and specificity of a milk pregnancy-associated glycoprotein ELISA test for pregnancy diagnosis between 23 and 27 days after insemination in Holstein dairy cows.

Pregnancy diagnosis using pregnancy-associated glycoprotein (PAG) ELISA technology in blood or milk samples is validated from 28 d after insemination in dairy cows. The objective of this study was to estimate the sensitivity (Se) and specificity (Sp) of a commercial milk PAG-based ELISA in Holstein dairy cows between 23 and 27 d after insemination. Milk samples (n = 268) from 257 Holstein dairy cows 23 to 27 d after AI were submitted for PAG ELISA testing. Pregnancy status was confirmed by either a second milk PAG ELISA test conducted between 28 and 50 d after insemination (n = 200) or transrectal ultrasonography performed between 28 and 59 d after insemination (n = 68). A Bayesian latent class model was used to compare the paired results from the test at 23 to 27 d after AI test to the reference test. The latent class model typically used for comparing 2 or more imperfect tests was extended to include the possibility of pregnancy loss between the 23 to 27 d test and the reference test. Informative priors for the probability of pregnancy loss, and for the Se and Sp of the PAG and ultrasonography reference tests were obtained from the scientific literature. Estimated median Se and Sp of the PAG ELISA test conducted between 23 and 27 d after AI were 0.98 (95% credible interval 0.93 to 1.0) and 0.98 (0.89 to 1.0), respectively, when using a standardized corrected optical density threshold of 0.15. Although the accuracy of the test under investigation was excellent, more data will be needed to confirm the optimal diagnostic cut point for PAG in milk for early pregnancy diagnosis in this time window. The optimal timing of pregnancy diagnosis will depend on herd-specific logistics and the action to be taken to re-inseminate nonpregnant cows.

How does salinity influence habitat selection and growth in juvenile American eels Anguilla rostrata?

The influence of salinity on habitat selection and growth in juvenile American eels Anguilla rostrata captured in four rivers across eastern Canada was assessed in controlled experiments in 2011 and 2012. Glass eels were first categorized according to their salinity preferences towards fresh (FW), salt (SW) or brackish water (BW) and the growth rate of each group of elvers was subsequently monitored in controlled FW and BW environments for 7 months. Most glass eels (78-89%) did not make a choice, i.e. they remained in BW. Salinity preferences were not influenced by body condition, although a possible role of pigmentation could not be ruled out. Glass eels that did make a choice displayed a similar preference for FW (60-75%) regardless of their geographic origin but glass eels from the St Lawrence Estuary displayed a significantly higher locomotor activity than those from other regions. Neither the salinity preferences showed by glass eels in the first experiment nor the rearing salinities appeared to have much influence on growth during the experiments. Elvers from Nova Scotia, however, reached a significantly higher mass than those from the St Lawrence Estuary thus supporting the hypothesis of genetically (or epigenetically) based differences for growth between A. rostrata from different origins. These results provide important ecological knowledge for the sustained exploitation and conservation of this threatened species.

Homing and group cohesion in Atlantic cod Gadus morhua revealed by tagging experiments.

Homing behaviour and group cohesion in Atlantic cod Gadus morhua from the northern Gulf of St Lawrence were studied based on tagging-recapture data from two periods, the 1980s and a recent period from 1996 to 2008. Two or more tags from a single tagging experiment were frequently recovered together in subsequent years. The null hypothesis was tested that the frequency of matching tag recoveries occurred by chance only through random mixing of tagged G. morhua before their recapture by the commercial fishery. The alternative hypothesis was that non-random, positive association (group cohesion) existed among tagged individuals that persisted through time and during migrations. Results show that the G. morhua population exhibits a homing behaviour, with temporal stability across seasons and years: 50% of recaptured fish in the recent period were caught <34 km from their mark site, even 3 years after release. In the 1980s, G. morhua were located at <10 km from their release site 1 year after tagging during summer and at <16 km during spring and autumn combined. Despite the increasing distance between the mark and recapture sites over time, the difference was not significant. In addition, occurrences of two or more tagged fish from the same release event that were caught together indicated a non-random association among individual fish for periods of one to several years and through migrations over several hundred kilometres. Hence G. morhua showed group cohesion in addition to site fidelity. These two interacting behaviours may be fundamental for the rebuilding and conservation of depleted fish stocks.

Estimates of the mortality and the duration of the trans-Atlantic migration of European eel Anguilla anguilla leptocephali using a particle tracking model.

Using Lagrangian simulations, based on circulation models over three different hydroclimatic periods in the last 45 years in the North Atlantic Ocean, the trans-Atlantic migration of the European eel Anguilla anguilla leptocephali was simulated via the passive drift of particles released in the spawning area. Three different behaviours were modelled: drifting at fixed depth, undergoing a vertical migration or choosing the fastest currents. Simulations included mortality hypotheses to estimate a realistic mean migration duration and relative survival of A. anguilla larvae. The mean migration duration was estimated as 21 months and the mortality rate as 3.8 per year, i.e. < 0.2% of A. anguilla larvae may typically survive the trans-Atlantic migration.

Differential effects of origin and salinity rearing conditions on growth of glass eels of the American eel Anguilla rostrata: implications for stocking programmes.

In this study, growth patterns were monitored in controlled fresh and brackish water (BW) conditions for 7 months during Anguilla rostrata glass eel and elver stages. Null hypotheses tested were that there is no significant difference in growth between glass eels (1) collected from two geographic regions typified by different sex ratios, (2) reared in fresh and BW and (3) due to origin x salinity interactions. It was found that young A. rostrata from Mira River (MR, Cape Breton, Nova Scotia, Canada, an area where both males and females occur) grew faster than those from Grande-Rivière-Blanche (Québec, Canada, an area where population are highly skewed towards females; 99-100%). Anguilla rostrata from both origins also grew faster in BW, although there was a trend for origin x salinity interactions whereby this effect was more pronounced for fish from the MR. The results support the hypothesis that salinity can influence growth patterns, as possibly can quantitative genetic differences between A. rostrata glass eels from different origins. Possible explanations for these patterns and potential consequences for sex determination and translocation programmes are discussed.

Anguilla rostrata glass eel migration and recruitment in the estuary and Gulf of St Lawrence.

This study describes catches of Anguilla rostrata glass eels and associated oceanographic conditions in the St Lawrence Estuary and Gulf. Ichthyoplankton survey data suggest that they enter the Gulf primarily in May, migrate at the surface at night, and disperse broadly once they have passed Cabot Strait. They arrive in estuaries beginning at about mid-June and through the month of July. Migration extends west up to Québec City, in the freshwater zone of the St Lawrence Estuary, 1000 km west of Cabot Strait. Anguilla rostrata glass eels travel between Cabot Strait and receiving estuaries at a straight-line ground speed of c. 10-15 km day(-1). Catches of fish per unit effort in estuaries in the St Lawrence system are much lower than those reported for the Atlantic coast of Canada. Low abundance of A. rostrata glass eels in the St Lawrence system may be due to cold surface temperatures during the migration period which decrease swimming capacity, long distances from the spawning ground to Cabot Strait and from Cabot Strait to the destination waters (especially the St Lawrence River), complex circulation patterns, and hypoxic conditions in bottom waters of the Laurentian Channel and the St Lawrence Estuary.

Mott transition, compressibility divergence, and the P-T phase diagram of layered organic superconductors: an ultrasonic investigation.

The phase diagram of the organic superconductor kappa-(BEDT-TTF)(2)Cu[N(CN)(2)]Cl has been investigated by ultrasonic velocity measurements under helium gas pressure. Different phase transitions were identified through several elastic anomalies characterized from isobaric and isothermal sweeps. Our data reveal two crossover lines that end on the critical point terminating the first-order Mott transition line. When the critical point is approached along these lines, we observe a dramatic anomaly of the sound velocity which is consistent with a diverging compressibility of the electronic degrees of freedom.

First-generation aortic endografts: analysis of explanted Stentor devices from the EUROSTAR Registry.

PURPOSE: To examine the structure and healing characteristics of chronically implanted Stentor endografts that were explanted due to migration, endoleak, thrombosis, or aneurysm expansion. METHODS: The devices were harvested following reoperation (n = 5) or autopsy (n = 1) with implantation times ranging from 13 to 53 months. Structural modifications to the metal components were examined using radiography, endoscopy, and magnetic resonance imaging (MRI). Specimens taken from components of the modular stent-grafts were examined histologically and with scanning electron microscopy (SEM) to assess healing behavior. Physical and chemical stability of the nitinol wires and woven polyester graft material was evaluated using SEM and electron spectroscopy for chemical analysis. RESULTS: Although the endografts were retrieved for a variety of reasons, they exhibited similar healing and structural modifications. The woven polyester sleeve showed evidence of yarn shifting and distortion, yarn damage, and filament breakage leading to the formation of openings in the fabric. The luminal surface endografts showed incomplete healing characterized by a poorly organized, nonadherent thrombotic matrix of variable thickness. Radiographic and endoscopic observations indicated that structural failure of the grafts, particularly in the main aortic component, was related to severe compaction and dislocation of the metallic frame due to suture breaks. Corrosion marks were observed on some nitinol wires in all devices. Chemical analysis and ion bombardment of the nitinol wires revealed that the surface concentrations of titanium and nickel were not homogenous. The first layer was composed of carbon or organic elements, followed by a stratum of highly oxidized titanium with a low nickel concentration; the titanium-nickel alloy lay beneath these layers. CONCLUSIONS: Although the materials selected for construction of endovascular grafts appears judicious, the assembly of these biomaterials into various interrelated structures within the device requires further improvement.

cAMP-dependent mobilization of intracellular Ca2+ stores by activation of ryanodine receptors in pancreatic beta-cells. A Ca2+ signaling system stimulated by the insulinotropic hormone glucagon-like peptide-1-(7-37).

Glucagon-like peptide-1 (GLP-1) is an intestinally derived insulinotropic hormone currently under investigation for use as a novel therapeutic agent in the treatment of type 2 diabetes mellitus. In vitro studies of pancreatic islets of Langerhans demonstrated that GLP-1 interacts with specific beta-cell G protein-coupled receptors, thereby facilitating insulin exocytosis by raising intracellular levels of cAMP and Ca2+. Here we report that the stimulatory influence of GLP-1 on Ca2+ signaling results, in part, from cAMP-dependent mobilization of ryanodine-sensitive Ca2+ stores. Studies of human, rat, and mouse beta-cells demonstrate that the binding of a fluorescent derivative of ryanodine (BODIPY FL-X ryanodine) to its receptors is specific, reversible, and of high affinity. Rat islets and BTC3 insulinoma cells are shown by reverse transcriptase polymerase chain reaction analyses to express mRNA corresponding to the type 2 isoform of ryanodine receptor-intracellular Ca2+ release channel (RYR2). Single-cell measurements of [Ca2+]i using primary cultures of rat and human beta-cells indicate that GLP-1 facilitates Ca2+-induced Ca2+ release (CICR), whereby mobilization of Ca2+ stores is triggered by influx of Ca2+ through L-type Ca2+ channels. In these cells, GLP-1 is shown to interact with metabolism of D-glucose to produce a fast transient increase of [Ca2+]i. This effect is reproduced by 8-Br-cAMP, but is blocked by a GLP-1 receptor antagonist (exendin-(9-39)), a cAMP antagonist ((Rp)-cAMPS), an L-type Ca2+ channel antagonist (nimodipine), an antagonist of the sarco(endo)plasmic reticulum Ca2+ ATPase (thapsigargin), or by ryanodine. Characterization of the CICR mechanism by voltage clamp analysis also demonstrates a stimulation of Ca2+ release by caffeine. These findings provide new support for a model of beta-cell signal transduction whereby GLP-1 promotes CICR by sensitizing intracellular Ca2+ release channels to the stimulatory influence of cytosolic Ca2+.

Expression of adenylyl cyclase subtypes in pancreatic beta-cells.

Activation of adenylyl cyclase by Gs-coupled receptors for insulinotropic hormones such as glucagon-like peptide-1 and pituitary adenylate cyclase-activating polypeptide plays a critical role in stimulating glucose-induced insulin secretion. Despite this important role of insulinotropic hormones in the regulation of insulin secretion, little is known about which of the multiple subtypes of adenylyl cyclase are expressed in beta-cells. Here we report the use of PCR primers designed to amplify all subtypes of adenylyl cyclase from cDNA prepared from human and rat islets and from insulin-secreting beta-cell lines. PCR products were cloned and sequenced to identify the subtypes of adenylyl cyclase amplified. Adenylyl cyclase types V and VI, known to couple to Galphas and Gbetagamma in the cAMP signaling pathway, account for all subtypes identified in human islets and INS-1 cells and the majority of subtypes in rat islets and HIT-T15 cells. These findings indicate that pancreatic beta-cells are particularly well suited to transmit signals via Gs-coupled receptors such as that for glucagon-like peptide-1.

Role of the striatum, cerebellum, and frontal lobes in the learning of a visuomotor sequence.

This study was designed to examine the role of the striatum, cerebellum, and frontal lobes in the implicit learning of a visuomotor sequence. The performance of patients with idiopathic Parkinson's disease (PD), with damage to the cerebellum, or with a circumscribed lesion to the frontal lobes was thus compared to that of separate groups of matched normal control subjects on an adapted version of the Repeated Sequence Test. This paradigm consists of a visual reaction-time task with a fixed embedded sequence of finger movements to be performed based on presentation of visual stimuli. Subjects received four blocks of trials (i.e., 40 presentations of a 10-item sequence) per day over 6 training days. Following the last experimental session, subjects were also given two tests measuring their declarative knowledge of the sequence. Only PD patients with a bilateral striatal-dysfunction or patients with lesions to the cerebellum failed to improve their performance in the last three training sessions, hence suggesting an impairment late in the acquisition process. Further analyses revealed that such impairment was mainly implicit in nature, and that it could not be ascribed to a general decline in cognitive functioning, to mood disturbances, or to the severity of the motor symptoms. By contrast, the level of declarative knowledge of the sequence did not differ between the three clinical groups and their respective groups of normal subjects. These findings suggest that, unlike declarative memory, the incremental acquisition of a new visuomotor skill depends upon the integrity of both the striatum and the cerebellum, but not of the frontal lobes.

Excessive fragmentary myoclonus: time of night and sleep stage distributions.

Excessive fragmentary myoclonus during sleep consists of high amounts of brief twitch-like movements occurring asynchronously and asymmetrically in different body areas and has been reported to occur in association with a number of sleep disorders. It was analyzed using a new technique of quantification, the fragmentary myoclonus index (FMI). The FMI exhibited high rates in all stages of sleep but with a somewhat lower frequency in slow wave sleep explaining, as well, a significantly lower rate in the first hour after onset compared to later hours. There was no evidence for greater sleep fragmentation or lighter sleep compared to a matched patient group in whom it had not been noted.

Cultured human keratinocytes cannot metabolize vitamin D3 to 25-hydroxyvitamin D3.

The metabolism of [3H]vitamin D3 was studied in cultured human keratinocytes (CHK). Intact CHK were incubated for 1, 6, 12, 24 and 48 h with [3H]vitamin D3 and the lipid soluble fractions from the media and cells were extracted by high-performance liquid chromatography (HPLC). Vitamin D3 and its metabolites, 25-OH-D3, 24,25(OH)2D3 and 1,25(OH)2D3 were added to the extracts, as markers, prior to HPLC. HPLC analysis of the lipid extracts did not reveal any monohydroxylated metabolites. CHK incubated for one hour with [3H]25-OH-D3 showed a 10 +/- 4% conversion to [3H]1,25(OH)2D3 whereas no conversion to [3H]1,25(OH)2D3 was observed in control CHKs that were boiled prior to incubation with [3H]25-OH-D3. These findings suggest that cultured neonatal keratinocytes are incapable of metabolizing vitamin D3 to 25-OH-D3.