Inhibitory interneurons play a critical role in the generation of gamma (20-50 Hz) oscillations, either by forming mutually inhibitory networks or as part of recurrent networks with pyramidal cells. A key property of fast spiking interneurons is their ability to generate brief spikes and high frequency spike trains with little accommodation. However, the role of their firing properties in network oscillations has not been tested in vivo. Studies in hippocampus in vitro have shown that high frequency spike doublets in interneurons play a key role in the long-range synchronization of gamma oscillations with little phase lag despite long axonal conduction delays. We generated a knockout (KO) mouse lacking Kv3.2 potassium channel subunits, where infragranular inhibitory interneurons lose the ability both to sustain high frequency firing and reliably generate high frequency spike doublets. We recorded cortical local field potentials in anesthetized and awake, restrained mice. Spontaneous activity of the KO and the wild-type (WT) showed similar content of gamma and slow (0.1-15 Hz) frequencies, but the KO showed a significantly larger decay of synchronization of gamma oscillations with distance. Coronal cuts in the cortex of WT mice decreased synchronization to values similar to the intact KO. The synchronization of the slow oscillation showed little decay with distance in both mice, and was largely reduced after coronal cuts. Our results show that the firing properties of inhibitory interneurons are critical for long range synchronization of gamma oscillations, and emphasize that intrinsic electrophysiological properties of single cells may play a key role in the spatiotemporal characteristics of network activity.

The Study to Explore Early Development (SEED), a multisite investigation addressing knowledge gaps in autism phenotype and etiology, aims to:(1) characterize the autism behavioral phenotype and associated developmental, medical, and behavioral conditions and (2) investigate genetic and environmental risks with emphasis on immunologic, hormonal, gastrointestinal, and sociodemographic characteristics. SEED uses a case-control design with population-based ascertainment of children aged 2-5 years with an autism spectrum disorder (ASD) and children in two control groups-one from the general population and one with non-ASD developmental problems. Data from parent-completed questionnaires, interviews, clinical evaluations, biospecimen sampling, and medical record abstraction focus on the prenatal and early postnatal periods. SEED is a valuable resource for testing hypotheses regarding ASD characteristics and causes.

[This corrects the article on p. e28532 in vol. 6.].

ABSTRACT: We report a case of profound neurologic and cardiovascular manifestations of tricyclic antidepressant intoxication following self-poisoning with multiple pharmaceuticals including amitriptyline in excess of 43 mg/kg, in a 51-year-old male. Institution of mechanical ventilation, volume expansion, systemic alkalinisation (pH 7.51), and intermittent bolus metaraminol resulted in QRS narrowing but failed to resolve the developed shock. One 100-ml bolus of 20% lipid emulsion followed by a further 400 ml over 30 min was administered with restoration of haemodynamic stability, thereby curtailing the need for ongoing vasopressor medications. Assayed blood levels were consistent with the 'lipid sink' being a major effecter in the observed improvement.

The large-conductance Ca(2+)-activated K(+)(BK) channel and its β-subunit underlie tuning in non-mammalian sensory or hair cells, whereas in mammals its function is less clear. To gain insights into species differences and to reveal putative BK functions, we undertook a systems analysis of BK and BK-Associated Proteins (BKAPS) in the chicken cochlea and compared these results to other species. We identified 110 putative partners from cytoplasmic and membrane/cytoskeletal fractions, using a combination of coimmunoprecipitation, 2-D gel, and LC-MS/MS. Partners included 14-3-3γ, valosin-containing protein (VCP), stathmin (STMN), cortactin (CTTN), and prohibitin (PHB), of which 16 partners were verified by reciprocal coimmunoprecipitation. Bioinformatics revealed binary partners, the resultant interactome, subcellular localization, and cellular processes. The interactome contained 193 proteins involved in 190 binary interactions in subcellular compartments such as the ER, mitochondria, and nucleus. Comparisons with mice showed shared hub proteins that included N-methyl-D-aspartate receptor (NMDAR) and ATP-synthase. Ortholog analyses across six species revealed conserved interactions involving apoptosis, Ca(2+) binding, and trafficking, in chicks, mice, and humans. Functional studies using recombinant BK and RNAi in a heterologous expression system revealed that proteins important to cell death/survival, such as annexinA5, γ-actin, lamin, superoxide dismutase, and VCP, caused a decrease in BK expression. This revelation led to an examination of specific kinases and their effectors relevant to cell viability. Sequence analyses of the BK C-terminus across 10 species showed putative binding sites for 14-3-3, RAC-α serine/threonine-protein kinase 1 (Akt), glycogen synthase kinase-3β (GSK3β) and phosphoinositide-dependent kinase-1 (PDK1). Knockdown of 14-3-3 and Akt caused an increase in BK expression, whereas silencing of GSK3β and PDK1 had the opposite effect. This comparative systems approach suggests conservation in BK function across different species in addition to novel functions that may include the initiation of signals relevant to cell death/survival.

The stem cell intrinsic model of self-renewal via asymmetric cell division (ACD) posits that fate determinants be partitioned unequally between daughter cells to either activate or suppress the stemness state. ACD is a purported mechanism by which hematopoietic stem cells (HSCs) self-renew, but definitive evidence of this cellular process remains open to conjecture. To address this issue, we chose 73 candidate genes that function within the cell polarity network to identify potential determinants that may concomitantly alter HSC fate whilst also exhibiting asymmetrical segregation at cell division. Initial gene expression profiles of polarity candidates showed high and differential expression in both HSCs and leukemia stem cells. Altered HSC fate was assessed by our established in vitro to in vivo screen on a sub-cohort of candidate polarity genes. This revealed six novel positive regulators of HSC function: Ap2a2, Gpsm2, Tmod1, Kif3a, Racgap1 and Ccnb1. Interestingly, live cell videomicroscopy of the endocytic protein AP2A2 shows instances of asymmetric segregation during hematopoietic stem/progenitor cell cytokinesis. These results contribute further evidence that ACD is functional in HSC self-renewal, suggests a role for Ap2a2 in HSC activity, and provides a unique opportunity to prospectively analyze progeny from HSC asymmetric divisions.

Procedural sedation has become widespread in emergency departments (ED) worldwide due to the ability to perform short turnaround noxious procedures beyond the confines of the operating theatre. We report one institution's experience with paediatric forearm fracture reduction and compare key time-based metrics for ED manipulation under procedural sedation (MUS), with traditional theatre-based manipulation under anaesthesia (MUA). All simple paediatric forearm fractures requiring manipulation before casting at Waikato hospital during the 2009 calendar year were studied. Time from presentation to fracture manipulation, procedure room occupancy, and hospital length of stay were recorded. Requirement for repeated intervention was additionally collated. Of 385 patients presenting with forearm fracture 108 underwent MUS and 66 MUA. Time to manipulation was shorter in the MUS group (58 plus or minus 38 minutes MUS vs. 558 plus or minus 368 minutes MUA; p<0.0001), as was hospital length of stay (139 plus or minus 70 minutes MUS vs 1452 plus or minus 544 minutes MUA; p<0.0001). No difference was observed in requirement for repeated intervention between groups (15% MUS vs 21% MUA; p=0.305). Manipulation of simple closed paediatric forearm fractures under procedural sedation was associated with lesser delay to reduction, and shorter hospital length of stay, compared with traditional manipulation under anaesthesia in the operating theatre.

Procedural sedation is commonly employed in the emergency department to assist in performance of noxious or invasive procedures. Numerous studies exist purporting the safety and efficacy of procedural sedation in North America and Australia, however, little data on sedation practice within New Zealand has been reported. We present one-year experience of all procedural sedations performed at Waikato Hospital. A prospective audit of all procedural sedations performed at the emergency department of Waikato hospital during the 2009 calendar year was conducted. Data abstraction included: indication for sedation, procedure duration, emergency department length of stay, required personnel, and procedural success, in addition to sedative agents employed and associated adverse events. 589 (276 paediatric, 313 adult) episodes of procedural sedation were available for analysis. Successful procedure performance was reported in 98% of paediatric cases and 88% of adult cases. Ketamine was the most commonly employed agent in the paediatric population (83.6%), with propofol the most frequently used in adults (99%). Procedural duration and emergency department length of stay was median 15 (IQR 10-25) minutes, and 122 (IQR 85-164) minutes respectively for the paediatric group, and median 15 (IQR 10-20) minutes and 124 (81-192) minutes for adult patients. Discharge rates for paediatric and adult patients were 87% and 52% respectively. Complication rates of procedural sedation for both groups was low. Procedural sedation appears both safe and effective in performance of time-limited noxious manipulations in a 'real-life' emergency department setting in New Zealand.

A large population of children and adults is potentially exposed to indoor environmental quality (IEQ) hazards in schools. Those with asthma are particularly at risk because IEQ-related hazards in school buildings can trigger asthma episodes. A multiagency consortium created and led by the Connecticut Department of Public Health has successfully implemented and continues to sustain the U.S. Environmental Protection Agency's (U.S. EPA's) Tools for Schools (TfS) program in the majority of Connecticut public schools. TfS is an action kit and program promoting a low-cost, problem-solving team approach to preventing IEQ hazards or improving IEQ. One key to the consortium's success is the array of services it provides to schools, including aggressive outreach and specialized training and consultation. The consortium is also a platform for launching other school IEQ initiatives. The authors present and analyze the consortium model and their efforts at evaluating the impact of TfS in Connecticut.

OBJECTIVE: UGT2B7 is a key member of the UDP-glucuronosyltransferase (UGT) family that participates in glucuronidation of endogenous compounds and pharmaceuticals. Much evidence suggests a large interindividual variability of UGT2B7-mediated glucuronidation, which is still unexplained by polymorphisms. We hypothesized that alternative splicing may be responsible for the variability in the UGT2B7 function. METHODS: We carried out a comprehensive scan for additional exons at this locus and discovered multiple alternative splicing events. We then determined transcript expression profiles across a large variety of human tissues and assessed some of these variants for their glucuronidation activity in human cells. RESULTS: In-depth analysis of the UGT2B7 gene structure led to the discovery of six novel exons. Kidney and liver samples presented the greatest enrichment of tissue-specific splicing, with 21 new UGT2B7 transcripts isolated. Furthermore, transcription from the proximal promoter (exon 1), associated with the active UGT2B7 mRNA isoform 1 (UGT2B7_v1), is most commonly observed in the gastrointestinal tract, whereas a distal promoter (exon 1a) induces the exclusion of the canonical exon 1 and is active in hormone-related tissues. We also showed that novel transcripts with alternative 3' ends could be translated into proteins lacking glucuronosyltransferase activity in human cells but acting as negative regulators on transferase activity when coexpressed with the active UGT2B7 protein. CONCLUSION: Our findings point toward a significant variability in structure, abundance, and tissue-specific UGT2B7 transcriptome, in addition to novel functions for UGT2B7-derived proteins, all of which may ensure the production of tissue-specific proteomes and functions.