Balloon valvuloplasty of valvular pulmonary stenosis in a neonatal foal.

In a 1-day old filly with a loud heart murmur, transthoracic echocardiography revealed right ventricular hypertrophy associated with severe pulmonary valvular stenosis and a transvalvular pressure gradient (between right ventricle and pulmonary artery) of 125 mmHg. Computed tomographic angiography confirmed the finding, with no evidence of other relevant concurrent abnormalities. Balloon valvuloplasty was performed using a single balloon technique. The foal recovered well from anaesthesia. Following the procedure, the right ventricle-pulmonary artery transvalvular gradient decreased to 38 mmHg. At follow up examinations after 1 month, 1 year and 2 years, the filly showed normal exercise capacity and echocardiography confirmed the persistent substantial improvement in the transvalvular outflow gradient.

Quantification of left atrial wall motion in healthy horses using two-dimensional speckle tracking.

INTRODUCTION: The mechanical function of the left atrium (LA) plays a pivotal role in modulation of left ventricular filling. Assessment of LA function might be a clinically useful prognostic tool for horses with mitral regurgitation or atrial fibrillation. However, the most accurate, reliable, and clinically useful methods to assess LA myocardial function are yet to be determined. The objective of this study was to describe the methods for quantification of LA wall motion using two-dimensional speckle tracking (2DST) echocardiography in healthy Warmblood horses to quantify measurement variability, to calculate reference intervals for 2DST variables, and to investigate their relationship to sex, age, body weight, and heart rate. ANIMALS, MATERIALS, AND METHODS: Twenty-six healthy Warmblood horses were included. 2DST analyses of LA wall motion were performed on digitally stored cine-loop recordings of a standardized right-parasternal four-chamber view focusing on the LA. Longitudinal strain, longitudinal strain rate, and time to peak LA contraction were measured to characterize LA contractile, reservoir, and conduit function. Intraobserver and interobserver measurement variability was quantified, and reference intervals were calculated. RESULTS: The coefficient of variation for intraobserver and interobserver measurement variability ranged between 2.0-11.1% and 5.1-15.4%, respectively, for global strain, strain rate, and time to peak LA contraction. Reference intervals for healthy Warmblood horses were reported. CONCLUSION: This study shows that 2DST is a feasible and reliable method to quantify LA wall motion throughout the cardiac cycle in healthy Warmblood horses. Further studies are required to establish the clinical value of 2DST for assessment of LA function.

Plasma homocysteine concentrations in healthy horses and horses with atrial fibrillation.

INTRODUCTION: Homocysteine (HCY) is an amino acid produced from methionine metabolism. Plasma homocysteine concentrations ([HCY]p) are elevated (>13 µmol/L) in people with atrial fibrillation (AF) and can predict the recurrence of AF after cardioversion. This study aimed to validate a commercially available human HCY assay for use in horses to develop reference intervals for [HCY]p and compare [HCY]p in healthy horses and horses with AF. ANIMALS: Healthy horses (n = 27) and horses with AF (n = 55, 34 of which were cardioverted using transvenous electrical cardioversion). MATERIALS AND METHODS: Blood samples were analysed for HCY using an automated enzyme-cycling assay (Homocysteine Cobas C, Integra, Roche) and creatinine (compensated Jaffe method). Assay linearity and precision were assessed, reference intervals calculated and [HCY]p and creatinine compared between groups. RESULTS: The assay was precise (coefficient of variation 1.6-4.3%, n = 10 repetitions) and provided linear results (r = 0.99 for spiked and natural samples) for a range of [HCY]p. The reference interval for [HCY]p was 1.5-7.8 µmol/L. The plasma concentration of homocysteine was 4.65 ± 1.5 µmol/L (mean ± standard deviation) in healthy horses and 4.65 ± 1.72 µmol/L in horses with AF (p=0.99); [HCY]p was not associated with recurrence of AF (n = 18, p=0.97). A weak, positive correlation between plasma creatinine and [HCY]p was detected (r = 0.295, p=0.008, r2 = 0.11). CONCLUSIONS: This assay allows precise measurement of [HCY]p in horses. Unlike in people, [HCY]p is not increased in horses with AF and cannot predict AF recurrence. This might be due to differences in the underlying pathological mechanisms of AF development in people and horses.

Echocardiographic Assessment of Left Ventricular Function in Healthy Horses and in Horses with Heart Disease Using Pulsed-Wave Tissue Doppler Imaging.

BACKGROUND: Assessment of left ventricular (LV) function by tissue Doppler imaging (TDI) is not well established in horses with heart disease. OBJECTIVES: To describe the use of pulsed-wave (PW) TDI for the assessment of LV function, establish reference intervals, investigate effects of mitral regurgitation (MR), aortic regurgitation (AR), and primary myocardial disease (MD), and provide proof of concept for the use of PW TDI in Warmblood horses with heart disease. ANIMALS: Thirty healthy horses, 38 horses with MR, 25 with AR, 8 with MD. METHODS: Echocardiograms were retrospectively analyzed. Reference intervals were calculated. PW TDI indices of healthy horses and horses with MR, AR, and MD were compared by one-way ANOVA and Dunnett's test. RESULTS: A complete set of PW TDI variables could be obtained in 94 of 101 horses. Variables corresponding to isovolumic intervals were most difficult to measure. Valvular regurgitation influenced variables describing isovolumic contraction and ejection. Horses with MD had significantly shortened ETm (-118.5 [-154.1 to -82.9] ms; mean difference [95% CI of difference of means]), increased PEPm /ETm (0.11 [0.05 to 0.17]), prolonged IMPm (0.28 [0.18 to 0.37]), increased S1 (8.9 [5.2 to 12.6] cm/s), and decreased E1 (-2.6 [-4.7 to -0.5] cm/s), Em (-14.2 [-19.9 to -8.5] cm/s), and Em /Am ratio (-1.6 [-2.6 to -0.6]). CONCLUSIONS AND CLINICAL IMPORTANCE: Pulsed-wave TDI might be useful for detection of LV dysfunction in horses with primary MD. The clinical value of TDI in horses with MR and AR remains uncertain.

Echocardiographic Assessment of Left Atrial Size and Function in Warmblood Horses: Reference Intervals, Allometric Scaling, and Agreement of Different Echocardiographic Variables.

BACKGROUND: Echocardiographic assessment of left atrial (LA) size and function in horses is not standardized. OBJECTIVES: The aim of this study was to establish reference intervals for echocardiographic indices of LA size and function in Warmblood horses and to provide proof of concept for allometric scaling of variables and for the clinical use of area-based indices. ANIMALS: Thirty-one healthy Warmblood horses and 91 Warmblood horses with a primary diagnosis of mitral regurgitation (MR) or aortic regurgitation (AR). METHODS: Retrospective study. Echocardiographic indices of LA size and function were measured and scaled to body weight (BWT). Reference intervals were calculated, the influence of BWT, age, and valvular regurgitation on LA size and function was investigated and agreement between different measurements of LA size was assessed. RESULTS: Allometric scaling of variables of LA size allowed for correction of differences in BWT. Indices of LA size documented LA enlargement with moderate and severe MR and AR, whereas most indices of LA mechanical function were not significantly altered by valvular regurgitation. Different indices of LA size were in fair to good agreement but still lead to discordant conclusions with regard to assessment of LA enlargement in individual horses. CONCLUSIONS AND CLINICAL IMPORTANCE: Allometric scaling of echocardiographic variables of LA size is advised to correct for differences in BWT among Warmblood horses. Assessment of LA dimensions should be based on an integrative approach combining subjective evaluation and assessment of multiple measurements, including area-based variables. The clinical relevance of indices of LA mechanical function remains unclear when used in horses with mitral or aortic regurgitation.

Disrupted in schizophrenia 1 (DISC1) L100P mutants have impaired activity-dependent plasticity in vivo and in vitro.

Major neuropsychiatric disorders are genetically complex but share overlapping etiology. Mice mutant for rare, highly penetrant risk variants can be useful in dissecting the molecular mechanisms involved. The gene disrupted in schizophrenia 1 (DISC1) has been associated with increased risk for neuropsychiatric conditions. Mice mutant for Disc1 display morphological, functional and behavioral deficits that are consistent with impairments observed across these disorders. Here we report that Disc1 L100P mutants are less able to reorganize cortical circuitry in response to stimulation in vivo. Molecular analysis reveals that the mutants have a reduced expression of PSD95 and pCREB in visual cortex and fail to adjust expression of such markers in response to altered stimulation. In vitro analysis shows that mutants have impaired functional reorganization of cortical neurons in response to selected forms of neuronal stimulation, but there is no altered basal expression of synaptic markers. These findings suggest that DISC1 has a critical role in the reorganization of cortical plasticity and that this phenotype becomes evident only under challenge, even at early postnatal stages. This result may represent an important etiological mechanism in the emergence of neuropsychiatric disorders.

Formative evaluation of ontology learning methods for entity discovery by using existing ontologies as reference standards.

OBJECTIVE: Developing a two-step method for formative evaluation of statistical Ontology Learning (OL) algorithms that leverages existing biomedical ontologies as reference standards. METHODS: In the first step optimum parameters are established. A 'gap list' of entities is generated by finding the set of entities present in a later version of the ontology that are not present in an earlier version of the ontology. A named entity recognition system is used to identify entities in a corpus of biomedical documents that are present in the 'gap list', generating a reference standard. The output of the algorithm (new entity candidates), produced by statistical methods, is subsequently compared against this reference standard. An OL method that performs perfectly will be able to learn all of the terms in this reference standard. Using evaluation metrics and precision-recall curves for different thresholds and parameters, we compute the optimum parameters for each method. In the second step, human judges with expertise in ontology development evaluate each candidate suggested by the algorithm configured with the optimum parameters previously established. These judgments are used to compute two performance metrics developed from our previous work: Entity Suggestion Rate (ESR) and Entity Acceptance Rate (EAR). RESULTS: Using this method, we evaluated two statistical OL methods for OL in two medical domains. For the pathology domain, we obtained 49% ESR, 28% EAR with the Lin method and 52% ESR, 39% EAR with the Church method. For the radiology domain, we obtain 87% ESA, 9% EAR using Lin method and 96% ESR, 16% EAR using Church method. CONCLUSION: This method is sufficiently general and flexible enough to permit comparison of any OL method for a specific corpus and ontology of interest.

Rethinking the genetic architecture of schizophrenia.

BACKGROUND: For many years, the prevailing paradigm has stated that in each individual with schizophrenia (SZ) the genetic risk is due to a combination of many genetic variants, individually of small effect. Recent empirical data are prompting a re-evaluation of this polygenic, common disease-common variant (CDCV) model. Evidence includes a lack of the expected strong positive findings from genome-wide association studies and the concurrent discovery of many different mutations that individually strongly predispose to SZ and other psychiatric disorders. This has led some to adopt a mixed model wherein some cases are caused by polygenic mechanisms and some by single mutations. This model runs counter to a substantial body of theoretical literature that had supposedly conclusively rejected Mendelian inheritance with genetic heterogeneity. Here we ask how this discrepancy between theory and data arose and propose a rationalization of the recent evidence base. METHOD: In light of recent empirical findings, we reconsider the methods and conclusions of early theoretical analyses and the explicit assumptions underlying them. RESULTS: We show that many of these assumptions can now be seen to be false and that the model of genetic heterogeneity is consistent with observed familial recurrence risks, endophenotype studies and other population-wide parameters. CONCLUSIONS: We argue for a more biologically consilient mixed model that involves interactions between disease-causing and disease-modifying variants in each individual. We consider the implications of this model for moving SZ research beyond statistical associations to pathogenic mechanisms.

Rural-urban differences in the presentation, management and survival of breast cancer in Western Australia.

From all women diagnosed with invasive breast cancer in 1999 in Western Australia, rural and urban women were compared with regard to mode of detection, tumour characteristics at presentation, diagnostic investigations, treatment and survival. Women from rural areas with breast cancer (n=206, 23%) were less likely to have open biopsy with frozen section (P<0.001), breast-conserving surgery (P<0.001), adjuvant radiotherapy (P=0.004) and hormonal therapy (P=0.03), and were less likely to be treated by a high caseload breast cancer surgeon (P<0.001). Adjusting for age and tumour characteristics, rural women had an increased likelihood of death within 5 years of breast cancer diagnosis (HR 1.62, 95% CI 1.10-2.38). This difference was not significant after adjustment for treatment factors (HR 1.36, 95% CI 0.90-2.04).

Intracellular ATP-sensitive K+ channels in mouse pancreatic beta cells: against a role in organelle cation homeostasis.

AIMS/HYPOTHESIS: ATP-sensitive K(+) (K(ATP)) channels located on the beta cell plasma membrane play a critical role in regulating insulin secretion and are targets for the sulfonylurea class of antihyperglycaemic drugs. Recent reports suggest that these channels may also reside on insulin-containing dense-core vesicles and mitochondria. The aim of this study was to explore these possibilities and to test the hypothesis that vesicle-resident channels play a role in the control of organellar Ca(2+) concentration or pH. METHODS: To quantify the subcellular distribution of the pore-forming subunit Kir6.2 and the sulfonylurea binding subunit SUR1 in isolated mouse islets and clonal pancreatic MIN6 beta cells, we used four complementary techniques: immunoelectron microscopy, density gradient fractionation, vesicle immunopurification and fluorescence-activated vesicle isolation. Intravesicular and mitochondrial concentrations of free Ca(2+) were measured in intact or digitonin-permeabilised MIN6 cells using recombinant, targeted aequorins, and intravesicular pH was measured with the recombinant fluorescent probe pHluorin. RESULTS: SUR1 and Kir6.2 immunoreactivity were concentrated on dense-core vesicles and on vesicles plus the endoplasmic reticulum/Golgi network, respectively, in both islets and MIN6 cells. Reactivity to neither subunit was detected on mitochondria. Glibenclamide, tolbutamide and diazoxide all failed to affect Ca(2+) uptake into mitochondria, and K(ATP) channel regulators had no significant effect on intravesicular free Ca(2+) concentrations or vesicular pH. CONCLUSIONS/INTERPRETATION: A significant proportion of Kir6.2 and SUR1 subunits reside on insulin-secretory vesicles and the distal secretory pathway in mouse beta cells but do not influence intravesicular ion homeostasis. We propose that dense-core vesicles may serve instead as sorting stations for the delivery of channels to the plasma membrane.

Unilateral nephrectomy as a treatment for renal trauma in a foal.

A 2-day-old Warmblood colt foal was referred for evaluation of progressive abdominal distension and lethargy. Haemoperitoneum was diagnosed and a ventral midline laparotomy revealed capsular rupture, sub capsular haematoma and haemorrhage of the left kidney. Unilateral nephrectomy was performed and the foal recovered uneventfully, with no reported complications 1 year later.

Dense core secretory vesicles revealed as a dynamic Ca(2+) store in neuroendocrine cells with a vesicle-associated membrane protein aequorin chimaera.

The role of dense core secretory vesicles in the control of cytosolic-free Ca(2+) concentrations ([Ca(2+)](c)) in neuronal and neuroendocrine cells is enigmatic. By constructing a vesicle-associated membrane protein 2-synaptobrevin.aequorin chimera, we show that in clonal pancreatic islet beta-cells: (a) increases in [Ca(2+)](c) cause a prompt increase in intravesicular-free Ca(2+) concentration ([Ca(2+)]SV), which is mediated by a P-type Ca(2+)-ATPase distinct from the sarco(endo) plasmic reticulum Ca(2+)-ATPase, but which may be related to the PMR1/ATP2C1 family of Ca(2+) pumps; (b) steady state Ca(2+) concentrations are 3-5-fold lower in secretory vesicles than in the endoplasmic reticulum (ER) or Golgi apparatus, suggesting the existence of tightly bound and more rapidly exchanging pools of Ca(2+); (c) inositol (1,4,5) trisphosphate has no impact on [Ca(2+)](SV) in intact or permeabilized cells; and (d) ryanodine receptor (RyR) activation with caffeine or 4-chloro-3-ethylphenol in intact cells, or cyclic ADPribose in permeabilized cells, causes a dramatic fall in [Ca(2+)](SV). Thus, secretory vesicles represent a dynamic Ca(2+) store in neuroendocrine cells, whose characteristics are in part distinct from the ER/Golgi apparatus. The presence of RyRs on secretory vesicles suggests that local Ca(2+)-induced Ca(2+) release from vesicles docked at the plasma membrane could participate in triggering exocytosis.

Contextual overlap and eyewitness suggestibility.

Studies of eyewitness suggestibility have traditionally used a paradigm that maximizes the extent to which the postevent interview overlaps with the witnessed event in terms of narrative content, narrative structure, and environmental context. The present study explored whether these dimensions of overlap contribute to people's tendency to confuse suggested details for those they have actually witnessed. We systematically manipulated the extent to which the postevent questionnaire overlapped with the witnessed event. Across two experiments, overlap in narrative content, narrative structure, or environmental context was not found to increase suggestibility effects, even though the manipulation did have other memory effects (e.g., it improved cued recall of the actual source of the suggestions, Experiment 2). These findings suggest that understanding the interaction between the structure and content of the objective context in which misinformation is encountered and various remembering contexts (e.g., recognition vs. recall) is important for advancing our understanding of source confusion in an eyewitness situation.

Source ROCs are (typically) curvilinear: comment on Yonelinas (1999).

On the basis of his assumption that recollection is a threshold process, A. P. Yonelinas (1999) predicted linear source-identification receiver operating characteristics (ROCs) and recently reported data that were consistent with this prediction. In this article, the authors present data showing curvilinear source-identification ROCs across various encoding and test conditions. On the basis of the source-monitoring framework (e.g., M. K. Johnson, S. Hashtroudi, & D. S. Lindsay, 1993), the authors argue that curvilinearity of source-identification ROCs is a result of differences in the qualitative characteristics of memories rather than simply the influence of undifferentiated familiarity as the dual-process model might suggest.

Functional analysis of secreted and transmembrane proteins critical to mouse development.

We describe the successful application of a modified gene-trap approach, the secretory trap, to systematically analyze the functions in vivo of large numbers of genes encoding secreted and membrane proteins. Secretory-trap insertions in embryonic stem cells can be transmitted to the germ line of mice with high efficiency and effectively mutate the target gene. Of 60 insertions analyzed in mice, one-third cause recessive lethal phenotypes affecting various stages of embryonic and postnatal development. Thus, secretory-trap mutagenesis can be used for a genome-wide functional analysis of cell signaling pathways that are critical for normal mammalian development and physiology.

Risk factors for and impact of online sexual solicitation of youth.

CONTEXT: Health care professionals, educators, and others are increasingly called upon to advise parents and policymakers about risks posed to children by Internet use. However, little scientific information exists on the experiences of children online. OBJECTIVE: To assess the risk factors surrounding online sexual solicitations of youth and distress due to solicitation. DESIGN, SETTING, AND PARTICIPANTS: Telephone survey (August 1999-February 2000) of a random sample of 1501 youth aged 10 through 17 years who were regular Internet users. MAIN OUTCOME MEASURES: Demographic and behavioral characteristics associated with solicitation risk and distress due to solicitation. RESULTS: Nineteen percent of youth who used the Internet regularly were the targets of unwanted sexual solicitation in the last year. Girls (P<.001), older teens (P =.005), troubled youth (P =.004), frequent Internet users (P =.01), chat room participants (P<.001), and those who communicated online with strangers (P<.001) were at greater risk. Twenty-five percent of the solicited youth reported high levels of distress after solicitation incidents. Risk of distress was more common among the younger youth (P =.005), those who received aggressive solicitations (the solicitor attempted or made offline contact) (P =.001), and those who were solicited on a computer away from their home (P =.001). CONCLUSIONS: Many young people who use the Internet encounter unwanted sexual overtures. Health care professionals, educators, and parents should be prepared to educate youth about how to respond to online sexual solicitations, including encouraging youth to disclose and report such encounters and to talk about them.

Defining brain wiring patterns and mechanisms through gene trapping in mice.

The search to understand the mechanisms regulating brain wiring has relied on biochemical purification approaches in vertebrates and genetic approaches in invertebrates to identify molecular cues and receptors for axon guidance. Here we describe a phenotype-based gene-trap screen in mice designed for the large-scale identification of genes controlling the formation of the trillions of connections in the mammalian brain. The method incorporates an axonal marker, which helps to identify cell-autonomous mechanisms in axon guidance, and has generated a resource of mouse lines with striking patterns of axonal labelling, which facilitates analysis of the normal wiring diagram of the brain. Studies of two of these mouse lines have identified an in vivo guidance function for a vertebrate transmembrane semaphorin, Sema6A, and have helped re-evaluate that of the Eph receptor EphA4.

Aging and reflective processes of working memory: binding and test load deficits.

It was hypothesized that age-related deficits in episodic memory for feature combinations (e.g., B. L. Chalfonte & M. K. Johnson, 1996) signal, in part, decrements in the efficacy of reflective component processes (e.g., M. K. Johnson, 1992) that support the short-term maintenance and manipulation of information during encoding (e.g., F. 1. M. Craik. R. G. Morris. & M. L. Gick, 1990; T. A. Salthouse, 1990). Consistent with this, age-related binding deficits in a working memory task were found in 2 experiments. Evidence for an age-related test load deficit was also found: Older adults had greater difficulty than young adults when tested on 2 features rather than 1, even when binding was not required. Thus, disruption of source memory in older adults may involve deficits in both encoding processes (binding deficits) and monitoring processes (difficulty accessing multiple features, evaluating them, or both).

fMRI evidence of age-related hippocampal dysfunction in feature binding in working memory.

Richly detailed memories for particular events depend on processes that bind individual features of experience together. Previous cognitive behavioral research indicates that older adults have more difficulty than young adults in conditions requiring feature binding. We used functional magnetic resonance imaging (fMRI) during a working memory task to identify neural substrates of this age-related deficit in feature binding. For young, but not older, adults there was greater activation in left anterior hippocampus on combination trials (remember objects together with their locations) than on trials in which participants were told to remember only which objects or only which locations occurred. The results provide neuroimaging evidence for an age-related hippocampal dysfunction in feature binding in working memory.