Large-scale cellular-resolution gene profiling in human neocortex reveals species-specific molecular signatures.

Although there have been major advances in elucidating the functional biology of the human brain, relatively little is known of its cellular and molecular organization. Here we report a large-scale characterization of the expression of ∼1,000 genes important for neural functions by in situ hybridization at a cellular resolution in visual and temporal cortices of adult human brains. These data reveal diverse gene expression patterns and remarkable conservation of each individual gene's expression among individuals (95%), cortical areas (84%), and between human and mouse (79%). A small but substantial number of genes (21%) exhibited species-differential expression. Distinct molecular signatures, comprised of genes both common between species and unique to each, were identified for each major cortical cell type. The data suggest that gene expression profile changes may contribute to differential cortical function across species, and in particular, a shift from corticosubcortical to more predominant corticocortical communications in the human brain.

Hierarchical organization of cortical and thalamic connectivity.

The mammalian cortex is a laminar structure containing many areas and cell types that are densely interconnected in complex ways, and for which generalizable principles of organization remain mostly unknown. Here we describe a major expansion of the Allen Mouse Brain Connectivity Atlas resource1, involving around a thousand new tracer experiments in the cortex and its main satellite structure, the thalamus. We used Cre driver lines (mice expressing Cre recombinase) to comprehensively and selectively label brain-wide connections by layer and class of projection neuron. Through observations of axon termination patterns, we have derived a set of generalized anatomical rules to describe corticocortical, thalamocortical and corticothalamic projections. We have built a model to assign connection patterns between areas as either feedforward or feedback, and generated testable predictions of hierarchical positions for individual cortical and thalamic areas and for cortical network modules. Our results show that cell-class-specific connections are organized in a shallow hierarchy within the mouse corticothalamic network.

Shared and distinct transcriptomic cell types across neocortical areas.

The neocortex contains a multitude of cell types that are segregated into layers and functionally distinct areas. To investigate the diversity of cell types across the mouse neocortex, here we analysed 23,822 cells from two areas at distant poles of the mouse neocortex: the primary visual cortex and the anterior lateral motor cortex. We define 133 transcriptomic cell types by deep, single-cell RNA sequencing. Nearly all types of GABA (γ-aminobutyric acid)-containing neurons are shared across both areas, whereas most types of glutamatergic neurons were found in one of the two areas. By combining single-cell RNA sequencing and retrograde labelling, we match transcriptomic types of glutamatergic neurons to their long-range projection specificity. Our study establishes a combined transcriptomic and projectional taxonomy of cortical cell types from functionally distinct areas of the adult mouse cortex.

A mesoscale connectome of the mouse brain.

Comprehensive knowledge of the brain's wiring diagram is fundamental for understanding how the nervous system processes information at both local and global scales. However, with the singular exception of the C. elegans microscale connectome, there are no complete connectivity data sets in other species. Here we report a brain-wide, cellular-level, mesoscale connectome for the mouse. The Allen Mouse Brain Connectivity Atlas uses enhanced green fluorescent protein (EGFP)-expressing adeno-associated viral vectors to trace axonal projections from defined regions and cell types, and high-throughput serial two-photon tomography to image the EGFP-labelled axons throughout the brain. This systematic and standardized approach allows spatial registration of individual experiments into a common three dimensional (3D) reference space, resulting in a whole-brain connectivity matrix. A computational model yields insights into connectional strength distribution, symmetry and other network properties. Virtual tractography illustrates 3D topography among interconnected regions. Cortico-thalamic pathway analysis demonstrates segregation and integration of parallel pathways. The Allen Mouse Brain Connectivity Atlas is a freely available, foundational resource for structural and functional investigations into the neural circuits that support behavioural and cognitive processes in health and disease.

A comparison of conventional vs automated digital Peer Assessment Rating scoring using the Carestream 3600 scanner and CS Model+ software system: A randomized controlled trial.

INTRODUCTION: A prospective randomized study was undertaken to compare conventional study model-based manual Peer Assessment Rating (PAR) scoring with computer-based automated scoring using scanned study models or intraoral scanning. METHODS: The sample consisted of 67 patients, mean age 15.03 (range 11-37) years. Sixty-seven patients underwent alginate impression-taking and intraoral scanning (CS 3600; Carestream Dental, Stuttgart, Germany) at a single appointment in a randomized order. For each patient, a weighted PAR score was calculated manually by a calibrated examiner using study models and a PAR ruler (conventional group), and automatically using Carestream Dental CS Model+ software and data from scanned study models (indirect digital group) or intraoral scans (direct digital group). All procedures were timed, and each patient completed a binary questionnaire relating to their experience. RESULTS: There were no significant differences between methods for calculated mean weighted PAR score (P = 0.68). Mean (standard deviation) chairside time for impression-taking was 5.35 (± 1.16) minutes and for intraoral scanning, 7.76 (± 2.76) minutes (P <0.05). Mean (standard deviation) times taken to calculate weighted PAR scores were 2.86 (± 0.96), 5.58 (± 2.33), and 4.58 (± 2.18) minutes for conventional, indirect digital, and direct digital groups, respectively (P >0.05). A total of 61 patients (91%) preferred intraoral scanning to impression-taking. CONCLUSIONS: Automated PAR scoring using cast study models or intraoral scanning is valid, though both methods take longer than conventional scoring. Patients prefer intraoral scanning to impression-taking. REGISTRATION: ClinicalTrials.gov (NCT03405961). PROTOCOL: The protocol was not published before study commencement.

Recent advances in molecular biomarkers for diabetes mellitus: a systematic review.

CONTEXT: Diabetes is a growing global metabolic epidemic. Current research is focussing on exploring how the biological processes and clinical outcomes of diabetes are related and developing novel biomarkers to measure these relationships, as this can subsequently improve diagnostic, therapeutic and management capacity. OBJECTIVE: The objective of this study is to identify the most recent advances in molecular biomarkers of diabetes and directions that warrant further research. METHODS: Using a systematic search strategy, the MEDLINE, CINAHL and OVID MEDLINE databases were canvassed for articles that investigated molecular biomarkers for diabetes. Initial selections were made based on article title, whilst final inclusion was informed by a critical appraisal of the full text of each article. RESULTS: The systematic search returned 246 records, of which 113 were unique. Following screening, 29 records were included in the final review. Three main research strategies (the development of novel technologies, broad biomarker panels, and targeted approaches) identified a number of potential biomarkers for diabetes including miR-126, C-reactive protein, 2-aminoadipic acid and betatrophin. CONCLUSION: The most promising research avenue identified is the detection and quantification of micro RNA. Further, the utilisation of functionalised electrodes as a means to detect biomarker compounds also warrants attention.

Does systemic administration of casein phosphopeptides affect orthodontic movement and root resorption in rats?

OBJECTIVES: To assess the potential effects of casein phosphopeptides (CPPs) on orthodontically induced iatrogenic root resorption (OIIRR) and orthodontic teeth movement. MATERIALS AND METHODS: Forty Wistar rats (aged 11 weeks) were randomly divided into experimental group (EG; n = 20) that received a diet supplemented with CPP and control group (CG; n = 20) devoid of diet supplement. A 150 g force was applied using nickel titanium (NiTi) coil that was bonded on maxillary incisors and extended unilaterally to a maxillary first molar. At Day 28, animals in both groups were euthanized. Volumetric assessment of root resorption craters and linear measurement of maxillary first molars movement were blindly examined using a micro-computed tomography scan. RESULTS: Nine rats were excluded from the experiment due to loss during general anesthesia or appliances' failure. Intra-operator reproducibility was high in both volumetric and linear measurements, 92.8 per cent and 98.5-97.6 per cent, respectively. The results reveal that dietary CPP has statistically insignificant effect on the overall OIIRR and orthodontic movement. CONCLUSIONS: CPP seems to have statistically insignificant effect on the volume of OIIRR and orthodontic movement in rats. A long-term study with larger sample size using a different concentration of CPP is required to clarify the dentoalveolar effect of CPP.

Soluble guanylate cyclase activation during ischemic injury in mice protects against postischemic inflammation at the mitochondrial level.

The aim was to determine whether treatment with BAY 60-2770, a selective activator of oxidized soluble guanylate cyclase (sGC), near the end of an ischemic event would prevent postischemic inflammation and mitochondrial dysfunction in wild-type (WT) and heme oxygenase-1 KO (HO-1(-/-)) mice. This protocol prevented increases in leukocyte rolling (LR) and adhesion (LA) to intestinal venules along with elevated TNFα and circulating neutrophil levels that accompany ischemia-reperfusion (I/R) in both animal models. We further hypothesized that a component of BAY 60-2770 treatment involves maintenance of mitochondrial membrane integrity during I/R. Measurements on isolated enterocytes of calcein fluorescence (mitochondrial permeability) and JC-1 fluorescence ratio (mitochondrial membrane potential) were reduced by I/R, indicating formation of mitochondrial permeability transition pores (mPTP). These effects were abrogated by BAY 60-2770 as well as cyclosporin A and SB-216763, which prevented mPTP opening and inhibited glycogen synthase kinase-3ß (GSK-3ß), respectively. Western blots of WT and HO-1(-/-) enterocytes indicated that GSK-3ß phosphorylation on Ser(9) (inhibitory site) was reduced by half following I/R alone (increased GSK-3ß activity) and increased by one-third (reduced GSK-3ß activity) following BAY 60-2770. Other investigators have associated phosphorylation of the GSK-3ß substrate cyclophilin D (pCyPD) with mPTP formation. We observed a 60% increase in pCyPD after I/R, whereas BAY 60-2770 treatment of sham and I/R groups reduced pCyPD by about 20%. In conclusion, selective activation of oxidized sGC of WT and HO-1(-/-) during ischemia protects against I/R-induced inflammation and preserves mucosal integrity in part by reducing pCyPD production and mPTP formation.

Conserved molecular signatures of neurogenesis in the hippocampal subgranular zone of rodents and primates.

The neurogenic potential of the subgranular zone (SGZ) of the hippocampal dentate gyrus is likely to be regulated by molecular cues arising from its complex heterogeneous cellular environment. Through transcriptome analysis using laser microdissection coupled with DNA microarrays, in combination with analysis of genome-wide in situ hybridization data, we identified 363 genes selectively enriched in adult mouse SGZ. These genes reflect expression in the different constituent cell types, including progenitor and dividing cells, immature granule cells, astrocytes, oligodendrocytes and GABAergic interneurons. Similar transcriptional profiling in the rhesus monkey dentate gyrus across postnatal development identified a highly overlapping set of SGZ-enriched genes, which can be divided based on temporal profiles to reflect maturation of glia versus granule neurons. Furthermore, we identified a neurogenesis-related gene network with decreasing postnatal expression that is highly correlated with the declining number of proliferating cells in dentate gyrus over postnatal development. Many of the genes in this network showed similar postnatal downregulation in mouse, suggesting a conservation of molecular mechanisms underlying developmental and adult neurogenesis in rodents and primates. Conditional deletion of Sox4 and Sox11, encoding two neurogenesis-related transcription factors central in this network, produces a mouse with no hippocampus, confirming the crucial role for these genes in regulating hippocampal neurogenesis.

Correlated gene expression and target specificity demonstrate excitatory projection neuron diversity.

The neocortex contains diverse populations of excitatory neurons segregated by layer and further definable by their specific cortical and subcortical projection targets. The current study describes a systematic approach to identify molecular correlates of specific projection neuron classes in mouse primary somatosensory cortex (S1), using a combination of in situ hybridization (ISH) data mining, marker gene colocalization, and combined retrograde labeling with ISH for layer-specific marker genes. First, we identified a large set of genes with specificity for each cortical layer, and that display heterogeneous patterns within those layers. Using these genes as markers, we find extensive evidence for the covariation of gene expression and projection target specificity in layer 2/3, 5, and 6, with individual genes labeling neurons projecting to specific subsets of target structures. The combination of gene expression and target specificity imply a great diversity of projection neuron classes that is similar to or greater than that of GABAergic interneurons. The covariance of these 2 phenotypic modalities suggests that these classes are both discrete and genetically specified.

The Hawlix: a simple and aesthetic prosthetic- orthodontic retainer.

This report introduces the Hawlix, a hybrid thermoformed and cold-cured retainer. It was developed to overcome the limitations that modified Hawley retainers possess when used in patients with bounded saddles, attributable to dental trauma or hypodontia. The Hawlix can be used as an intermediate retainer while a patient is waiting to receive restorative treatment. Some of the advantages of the Hawlix over the modified Hawley retainer include: superior aesthetics, cost-effectiveness, ease of manufacture, adequate resistance to fracture, preservation of a residual ridge, and precise space maintenance.

J.G. Crowther's War: Institutional strife at the BBC and British Council.

Science writer, historian and administrator J.G. Crowther (1899-1983) had an uneasy relationship with the BBC during the 1920s and 1930s, and was regarded with suspicion by the British security services because of his left politics. Nevertheless the Second World War saw him working for 'establishment' institutions. He was closely associated with the BBC's Overseas Service and employed by the British Council's Science Committee. Both organizations found Crowther useful because of his wide, international knowledge of science and scientists. Crowther's political views, and his international aspirations for the British Council's Science Committee, increasingly embroiled him in an institutional conflict with the Royal Society and with its president, Sir Henry Dale, who was also chairman of the British Council's Science Committee. The conflict centred on the management of international scientific relations, a matter close Crowther's heart, and to Dale's. Dale considered that the formal conduct of international scientific relations was the Royal Society's business rather than the British Council's. Crowther disagreed, and eventually resigned from the British Council Science Committee in 1946. The article expands knowledge of Crowther by drawing on archival documents to elucidate a side of his career that is only lightly touched on in his memoirs. It shows that 'Crowther's war' was also an institutional war between the Science Committee of the British Council and the Royal Society. Crowther's unhappy experience of interference by the Royal Society plausibly accounts for a retreat from his pre-war view that institutional science should plan and manage BBC science broadcasts.

Denmark's comparative position regarding health status, healthcare provision, self-management and social support: Diabetes Attitudes, Wishes and Needs second study (DAWN2).

AIMS: The aim of this study was to benchmark the Danish sample of the second Diabetes, Attitudes, Wishes and Needs (DAWN2) study with the global average in order to determine Denmark's comparative position for health status, healthcare provision, self-management and social support from the perspective of people with diabetes, family members of people with diabetes and healthcare professionals. METHODS: A total of 502 Danish people with diabetes (PWD), 122 adult family members of people with diabetes (FM) and 283 healthcare professionals (HCPs) participated in the study. Data on healthcare provision and physical and psychosocial wellbeing were collected from the 17 participating countries. RESULTS: Psychological wellbeing was higher among Danish PWD; conversely, self-management behaviour of PWD ranked below the global average. A substantial gap was found in the perceptions of PWD and HCPs regarding the extent to which healthcare provision was deemed person-centred. The gap was found to be larger, however, when looking at the global data. Danish FM reported higher education participation and satisfaction rates as well as lower distress than the global average, but there appears to be an untapped potential when it comes to converting education participation of FM into social support for PWD. CONCLUSIONS: Our findings suggest that PWD in Denmark rank above the global average on measures of psychological wellbeing, despite psychological wellbeing being under-prioritised by HCP. However, there is room for improvement when it comes to self-management behaviours. Special attention is needed to address this issue without compromising the psychological wellbeing of the PWD.

The association between interpersonal problems and treatment outcome in the eating disorders: A systematic review.

OBJECTIVE: To review systematically the eating disorder literature in order to examine the association between pre-treatment interpersonal problems and treatment outcome in people diagnosed with an eating disorder. METHODS: Six relevant databases were searched for studies in which interpersonal problems prior to treatment were examined in relation to treatment outcome in patients diagnosed with anorexia nervosa (AN), bulimia nervosa (BN) or eating disorders not otherwise specified (EDNOS). RESULTS: Thirteen studies were identified (containing 764 AN, 707 BN and 48 EDNOS). The majority of studies indicated that interpersonal problems at the start of therapy were associated with a detrimental treatment outcome. CONCLUSIONS: Individuals with a binge/purge-type of eating disorder may be particularly vulnerable to interpersonal issues and these issues may lead to poorer treatment recovery by reducing the individual's ability to engage in the treatment process on a functional level. The clinical and research implications are discussed.

Physical properties of root cementum: Part 25. Extent of root resorption after the application of light and heavy buccopalatal jiggling forces for 12 weeks: A microcomputed tomography study.

INTRODUCTION: The aims of this study were to evaluate with microcomputed tomography the orthodontically induced inflammatory root resorption in premolars caused by buccopalatal jiggling movement with light and heavy forces and to compare it with the resorption caused by equivalent but continuous buccal forces. METHODS: The sample consisted of 60 maxillary first premolars collected from 30 patients (15 girls, 15 boys; ages, 13-18 years) who required orthodontic treatment with extractions. They were divided into 3 groups of 10 patients. Light (25 g) or heavy (225 g) buccal tipping orthodontic forces were randomly assigned on the maxillary right or left quadrant with either continuous buccal (positive controls) or buccopalatal jiggling forces for 12 weeks. At the end of the experimental period, the teeth were carefully extracted and processed for 3-dimensional imaging and volumetric evaluations of resorption craters. Data were analyzed with Wilcoxon signed rank tests. RESULTS: There was no statistically significant difference between positive control light (P = 0.0173) and heavy (P = 0.0173) continuous forces and jiggling forces for both force magnitudes. However, statistically significant differences were observed between heavy and light jiggling forces (P = 0.038), with heavy jiggling forces causing greater total root resorption than light jiggling forces. CONCLUSIONS: Light and heavy jiggling forces in the buccopalatal direction did not cause significantly different amounts of root resorption when compared with continuous forces of the same magnitude. On the other hand, light jiggling forces resulted in less root resorption than heavy jiggling forces.

The Allen Brain Atlas: 5 years and beyond.

The Allen Brain Atlas, a Web-based, genome-wide atlas of gene expression in the adult mouse brain, was an experiment on a massive scale. The development of the atlas faced a combination of great technical challenges and a non-traditional open research model, and it encountered many hurdles on the path to completion and community adoption. Having overcome these challenges, it is now a fundamental tool for neuroscientists worldwide and has set the stage for the creation of other similar open resources. Nevertheless, there are many untapped opportunities for exploration.

ß1-integrin is essential for vasoregulation and smooth muscle survival in vivo.

OBJECTIVE: Integrins contribute to vascular morphogenesis through regulation of adhesion and assembly of the extracellular matrix. However, the role of ß1-integrin in the mature vascular wall is less clear. APPROACH AND RESULTS: We sought to determine the function of ß1-integrin in mature smooth muscle cells in vivo using a loss of function approach by crossing a tamoxifen-inducible sm22αCre line to a floxed ß1-integrin transgenic line. Adult mice lacking smooth muscle ß1-integrin survived only 10 weeks post induction. The deletion of ß1-integrin resulted in profound loss of vasomotor control. Histological analysis revealed progressive fibrosis in arteries with associated apoptosis of smooth muscle cells, which was not rescued by adventitial stem cells. Smooth muscle cell apoptosis was detected in arteries with dead cells replaced primarily by collagen. Despite the catastrophic effects on vascular smooth muscle, the deleted visceral smooth muscle remained viable with the exception of a short portion of the colon, indicating that vascular but not visceral smooth muscle is particularly sensitive to changes in ß1-integrin. CONCLUSIONS: This study reveals an essential function of ß1-integrin in the maintenance of vasomotor control and highlights a critical role for ß1-integrin in vascular, but not visceral, smooth muscle survival.

Ultrasound imaging of the trapeziometacarpal articular cavity to investigate the presence of intraarticular gas bubbles after chiropractic manipulation.

OBJECTIVE: The purpose of this study was to investigate the presence of intraarticular gas bubbles in the trapeziometacarpal joint cavity after chiropractic manipulation with audible cavitation and to assess the state of the gas bubbles after a 20-minute refractory period. METHODS: This investigation included 18 asymptomatic male and female participants between the ages of 21 and 26 years. High-resolution (15 MHz) sonograms of the trapeziometacarpal articular cavity were obtained by an experienced musculoskeletal ultrasonographer at 3 intervals: premanipulation, within 30 seconds postmanipulation, and at 20 minutes postmanipulation. The sonograms were saved as digital copies for subsequent reports that were correlated with reports compiled during dynamic visualization of the articular cavity. Data were extracted from the reports for analysis. RESULTS: The premanipulative sonograms showed that 27.78% of joints contained minute gas bubbles, also known as microcavities, within the synovial fluid before the joint was manipulated. The remaining 72.22% of joints contained no intraarticular microcavities. All of the postmanipulative sonograms revealed numerous large conspicuous gas bubbles within the synovial fluid. The postrefractory sonograms showed that, in 66.66% of the synovial fluid, gas bubbles were still visible, whereas the remaining 33.34% had no presence of gas bubbles or microcavities, and the synovial fluid had returned to its premanipulative state. CONCLUSION: The findings of this study suggest that synovial fluid may contain intraarticular microcavities even before a manipulation is performed. Numerous large intraarticular gas bubbles are formed during manipulation due to cavitation of the synovial fluid and were observed in the absence of an axial distractive load at the time of imaging. In most cases, these gas bubbles remained within the joint for longer than 20 minutes.

Preconditioning with soluble guanylate cyclase activation prevents postischemic inflammation and reduces nitrate tolerance in heme oxygenase-1 knockout mice.

Previously we have shown that, unlike wild-type mice (WT), heme oxygenase-1 knockout (HO-1-/-) mice developed nitrate tolerance and were not protected from inflammation caused by ischemia-reperfusion (I/R) when preconditioned with a H2S donor. We hypothesized that stimulation (with BAY 41-2272) or activation (with BAY 60-2770) of soluble guanylate cyclase (sGC) would precondition HO-1-/- mice against an inflammatory effect of I/R and increase arterial nitrate responses. Intravital fluorescence microscopy was used to visualize leukocyte rolling and adhesion to postcapillary venules of the small intestine in anesthetized mice. Relaxation to ACh and BAY compounds was measured on superior mesenteric arteries isolated after I/R protocols. Preconditioning with either BAY compound 10 min (early phase) or 24 h (late phase) before I/R reduced postischemic leukocyte rolling and adhesion to sham control levels and increased superior mesenteric artery responses to ACh, sodium nitroprusside, and BAY 41-2272 in WT and HO-1-/- mice. Late-phase preconditioning with BAY 60-2770 was maintained in HO-1-/- and endothelial nitric oxide synthase knockout mice pretreated with an inhibitor (dl-propargylglycine) of enzymatically produced H2S. Pretreatment with BAY compounds also prevented the I/R increase in small intestinal TNF-α. We speculate that increasing sGC activity and related PKG acts downstream to H2S and disrupts signaling processes triggered by I/R in part by maintaining low cellular Ca²âº. In addition, BAY preconditioning did not increase sGC levels, yet increased the response to agents that act on reduced heme-containing sGC. Collectively these actions would contribute to increased nitrate sensitivity and vascular function.

The relationship between bone, hemopoietic stem cells, and vasculature.

A large body of evidence suggests hemopoietic stem cells (HSCs) exist in an endosteal niche close to bone, whereas others suggest that the HSC niche is intimately associated with vasculature. In this study, we show that transplanted hemopoietic stem and progenitor cells (HSPCs) home preferentially to the trabecular-rich metaphysis of the femurs in nonablated mice at all time points from 15 minutes to 15 hours after transplantation. Within this region, they exist in an endosteal niche in close association with blood vessels. The preferential homing of HSPCs to the metaphysis occurs rapidly after transplantation, suggesting that blood vessels within this region may express a unique repertoire of endothelial adhesive molecules. One candidate is hyaluronan (HA), which is highly expressed on the blood vessel endothelium in the metaphysis. Analysis of the early stages of homing and the spatial dis-tribution of transplanted HSPCs at the single-cell level in mice devoid of Has3-synthesized HA, provides evidence for a previously undescribed role for HA expressed on endothelial cells in directing the homing of HSPCs to the metaphysis.