Preparation and characterization of negatively charged poly(lactic-co-glycolic acid) microspheres.
Negatively charged poly(lactic-co-glycolic acid) (PLGA) microspheres encapsulated with hydrophilic drugs have been successfully prepared by a solid-in-oil-in-water (s/o/w) solvent evaporation method in the presence of anionic surfactants, sodium dodecyl sulfate (SDS), and dioctyl sodium sulfosuccinate (DSS), and nonionic surfactant polyvinyl alcohol (PVA). The effects of microencapsulation methods, surfactants types, and surfactant concentrations on the properties of microspheres were studied. Amoxicillin (AMX) was chosen as a hydrophilic model drug, and its encapsulation efficiency (EE) and in vitro release profiles were measured. The s/o/w method achieved higher EE of 40% in PLGA microspheres using surfactant SDS compared with the conventional water-in-oil-in-water (w/o/w) method (about 2%). Triphasic release profiles were observed for all PLGA microspheres (s/o/w) with slight drug burst, a slow diffusion-controlled release within the period of about 7 days and followed by the degradation-controlled sustained release for further 30 days. Smaller particle size and surface charge were achieved for s/o/w method than w/o/w method using the same anionic surfactants, and smooth surface and less porous interior matrix. The s/o/w method effectively encapsulated AMX into anionic PLGA microspheres using anionic surfactants, and these negatively charged PLGA microspheres represented an attractive approach for the controlled release of hydrophilic drugs.


Quantification of the pathological response and fate in the lung and pleura of chrysotile in combination with fine particles compared to amosite-asbestos following short-term inhalation exposure.
The marked difference in biopersistence and pathological response between chrysotile and amphibole asbestos has been well documented. This study is unique in that it has examined a commercial chrysotile product that was used as a joint compound. The pathological response was quantified in the lung and translocation of fibers to and pathological response in the pleural cavity determined. This paper presents the final results from the study. Rats were exposed by inhalation 6 h/day for 5 days to a well-defined fiber aerosol. Subgroups were examined through 1 year. The translocation to and pathological response in the pleura was examined by scanning electron microscopy and confocal microscopy (CM) using noninvasive methods. The number and size of fibers was quantified using transmission electron microscopy and CM. This is the first study to use such techniques to characterize fiber translocation to and the response of the pleural cavity. Amosite fibers were found to remain partly or fully imbedded in the interstitial space through 1 year and quickly produced granulomas (0 days) and interstitial fibrosis (28 days). Amosite fibers were observed penetrating the visceral pleural wall and were found on the parietal pleural within 7 days postexposure with a concomitant inflammatory response seen by 14 days. Pleural fibrin deposition, fibrosis, and adhesions were observed, similar to that reported in humans in response to amphibole asbestos. No cellular or inflammatory response was observed in the lung or the pleural cavity in response to the chrysotile and sanded particles (CSP) exposure. These results provide confirmation of the important differences between CSP and amphibole asbestos.


Children's coping strategies for chemotherapy-induced nausea and vomiting.
To identify anticipatory, acute, and delayed chemotherapy-induced nausea and vomiting (CINV) frequency and coping strategies used among pediatric patients with cancer. Prospective, cohort design. A pediatric teaching hospital in the southern United States. A convenience sample of 40 children aged 7-12 years scheduled to receive either moderately emetic chemotherapy or highly emetic chemotherapy for cancer treatment. Children completed the Adapted Rhodes Index of Nausea and Vomiting for Pediatrics and the Kidcope-Younger Version. CINV and coping strategies. CINV occurred during the anticipatory, acute, and delayed times, with the highest frequency occurring during the delayed time. The most frequently used coping strategies were distraction and wishful thinking, whereas the most effective strategies were social support and distraction. No statistically significant differences were observed in the frequency or efficacy of coping strategies over time. CINV occurs throughout chemotherapy treatment. The most efficacious coping strategies included active and passive coping, with active coping strategies being more effective. Nurses should recognize that CINV occurs at all points of chemotherapy treatment. Nurses can assist children in developing active coping strategies to manage their CINV.


Dynamics of brain networks in the aesthetic appreciation.
Neuroimage experiments have been essential for identifying active brain networks. During cognitive tasks as in, e.g., aesthetic appreciation, such networks include regions that belong to the default mode network (DMN). Theoretically, DMN activity should be interrupted during cognitive tasks demanding attention, as is the case for aesthetic appreciation. Analyzing the functional connectivity dynamics along three temporal windows and two conditions, beautiful and not beautiful stimuli, here we report experimental support for the hypothesis that aesthetic appreciation relies on the activation of two different networks, an initial aesthetic network and a delayed aesthetic network, engaged within distinct time frames. Activation of the DMN might correspond mainly to the delayed aesthetic network. We discuss adaptive and evolutionary explanations for the relationships existing between the DMN and aesthetic networks and offer unique inputs to debates on the mind/brain interaction.


MEF2B-Nox1 signaling is critical for stretch-induced phenotypic modulation of vascular smooth muscle cells.
Blood vessel hemodynamics have profound influences on function and structure of vascular cells. One of the main mechanical forces influencing vascular smooth muscle cells (VSMC) is cyclic stretch (CS). Increased CS stimulates reactive oxygen species (ROS) production in VSMC, leading to their dedifferentiation, yet the mechanisms involved are poorly understood. This study was designed to test the hypothesis that pathological CS stimulates NADPH oxidase isoform 1 (Nox1)-derived ROS via MEF2B, leading to VSMC dysfunction via a switch from a contractile to a synthetic phenotype. Using a newly developed isoform-specific Nox1 inhibitor and gene silencing technology, we demonstrate that a novel pathway, including MEF2B-Nox1-ROS, is upregulated under pathological stretch conditions, and this pathway promotes a VSMC phenotypic switch from a contractile to a synthetic phenotype. We observed that CS (10% at 1 Hz) mimicking systemic hypertension in humans increased Nox1 mRNA, protein levels, and enzymatic activity in a time-dependent manner, and this upregulation was mediated by MEF2B. Furthermore, we show that stretch-induced Nox1-derived ROS upregulated a specific marker for synthetic phenotype (osteopontin), whereas it downregulated classical markers for contractile phenotype (calponin1 and smoothelin B). In addition, our data demonstrated that stretch-induced Nox1 activation decreases actin fiber density and augments matrix metalloproteinase 9 activity, VSMC migration, and vectorial alignment. These results suggest that CS initiates a signal through MEF2B that potentiates Nox1-mediated ROS production and causes VSMC to switch to a synthetic phenotype. The data also characterize a new Nox1 inhibitor as a potential therapy for treatment of vascular dysfunction in hypertension.


The Use of Helplines and Telehealth Support in Aotearoa/New Zealand During COVID-19 Pandemic Control Measures: A Mixed-Methods Study.
Early evidence suggests that the COVID-19 pandemic and associated interventions have affected mental well-being and associated health service use. the aim of this study was to examine the effect of the COVID-19 pandemic and associated public health measures on helpline and telehealth service demand. the study utilized a mixed methods research design. Segmented regression analyses were used first to identify changes in patterns of demand for Aotearoa/New Zealand national helplines (n = 11) from January 2020 until the end of March 2021. Thematic analysis of 23 in-depth interviews was used next to explore the reasons behind the quantitative findings from the perspective of various organizational stakeholders. the data from 1,244,293 Aotearoa/New Zealand national helplines' contacts between January 2020 and March 2021 showed a non-significant (1.4%) upward trend for the full range of observations. Throughout this period, a peak and trough pattern was observed. Significant demand increases were observed in anticipation of containment measures (12.4% increase from January to March 2020) and significant demand decreases coincided with relaxation of restrictions (6.9% decrease from April to June 2020). There were spikes in demand during public health interventions (i.e., mental health promotion, introduction of new helpline services) and regional lockdowns, but these did not result in significant changes in trends. In general, the demand for helplines stabilized at a new higher level. Most of the contacts occurred by telephone calls. Contacts by other methods (webchat, text, email) have shown higher uptake during the periods of lockdowns. Quantitative-qualitative data triangulation showed that youth and populations who were disproportionally negatively affected by unstable economic conditions and underemployment made more frequent contacts. Providers emphasized that increased demand could be viewed positively as a successful outcome of public health messaging; however, greater capacity is needed to better serve higher demand. COVID-19, related interventions, and measures of control were associated with an increase in contacts to helplines. However, the extent of the demand increases was lower than observed internationally. Moreover, in Aotearoa/New Zealand the reasons for increases in demand were often beyond the COVID-19 pandemic and measures of control.


Genetic relatedness reveals total population size of white sharks in eastern Australia and New Zealand.
Conservation concerns exist for many sharks but robust estimates of abundance are often lacking. Improving population status is a performance measure for species under conservation or recovery plans, yet the lack of data permitting estimation of population size means the efficacy of management actions can be difficult to assess, and achieving the goal of removing species from conservation listing challenging. For potentially dangerous species, like the white shark, balancing conservation and public safety demands is politically and socially complex, often leading to vigorous debate about their population status. This increases the need for robust information to inform policy decisions. We developed a novel method for estimating the total abundance of white sharks in eastern Australia and New Zealand using the genetic-relatedness of juveniles and applying a close-kin mark-recapture framework and demographic model. Estimated numbers of adults are small (ca. 280-650), as is total population size (ca. 2,500-6,750). However, estimates of survival probability are high for adults (over 90%), and fairly high for juveniles (around 73%). This represents the first direct estimate of total white shark abundance and survival calculated from data across both the spatial and temporal life-history of the animal and provides a pathway to estimate population trend.


SURVEY OF ARCTIC ALASKAN WILDLIFE FOR INFLUENZA A ANTIBODIES: LIMITED EVIDENCE FOR EXPOSURE OF MAMMALS.
Influenza A viruses (IAVs) are maintained in wild waterbirds and have the potential to infect a broad range of species, including wild mammals. The Arctic Coastal Plain of Alaska supports a diverse suite of species, including waterfowl that are common hosts of IAVs. Mammals co-occur with geese and other migratory waterbirds during the summer breeding season, providing a plausible mechanism for interclass transmission of IAVs. To estimate IAV seroprevalence and identify the subtypes to which geese, loons, Arctic foxes ( Vulpes lagopus), caribou ( Rangifer tarandus), and polar bears ( Ursus maritimus) are potentially exposed, we used a blocking enzyme-linked immunosorbent assay (bELISA) and a hemagglutination inhibition (HI) assay to screen for antibodies to IAVs in samples collected during spring and summer of 2012-16. Apparent IAV seroprevalence using the bELISA was 50.3% in geese (range by species: 46-52.8%), 9% in loons (range by species: 3-20%), and 0.4% in Arctic foxes. We found no evidence for exposure to IAVs in polar bears or caribou by either assay. Among geese, we estimated detection probability from replicate bELISA analyses to be 0.92 and also found good concordance (>85%) between results from bELISA and HI assays, which identified antibodies reactive to H1, H6, and H9 subtype IAVs. In contrast, the HI assay detected antibodies in only one of seven loon samples that were positive by bELISA; that sample had low titers to both H4 and H5 IAV subtypes. Our results provide evidence that a relatively high proportion of waterbirds breeding on the Arctic Coastal Plain are exposed to IAVs, although it is unknown whether such exposure occurs locally or on staging or wintering grounds. In contrast, seroprevalence of IAVs in concomitant Arctic mammals is apparently low.


Nitric oxide synthase isoforms play distinct roles during acute peritonitis.
Acute peritonitis is the most frequent complication of peritoneal dialysis (PD). Increased nitric oxide (NO) release by NO synthase (NOS) isoforms has been implicated in acute peritonitis, but the role played by the NOS isoforms expressed in the peritoneum is unknown. We investigated the structural and functional consequences of acute peritonitis induced by LPS in wild-type (WT) mice versus knockout mice (KO) for the endothelial NOS (eNOS), the inducible NOS (iNOS) or the neuronal NOS (nNOS). The level of NO metabolites (NOx) in the dialysate was maximal 18 h after LPS injection. LPS induced a significant increase in the transport of small solutes and decreased ultrafiltration in WT mice. These changes, which occurred without vascular proliferation, were paralleled by the upregulation of nNOS and eNOS, and the induction of iNOS. The transport modifications induced by LPS were significantly reversed in eNOS KO mice, but not modified in mice lacking iNOS or nNOS. In contrast, the increase of dialysate NOx was abolished in iNOS KO mice and significantly reduced in eNOS KO mice, but left unchanged in mice lacking nNOS. Mice lacking iNOS also showed more severe inflammatory changes, and a trend towards increased mortality following LPS. These data demonstrate specific roles for NOS isoforms in the peritoneal membrane and suggest that selective eNOS inhibition may improve peritoneal transport during acute peritonitis.


A Micro-Electrode Array device coupled to a laser-based system for the local stimulation of neurons by optical release of glutamate.
Optical stimulation is a promising approach to investigate the local dynamic responses of cultured neurons. In particular, flash photolysis of caged compounds offers the advantage of allowing the rapid change of concentration of either extracellular or intracellular molecules, such as neurotransmitters or second messengers, for the stimulation or modulation of neuronal activity. We describe here the use of an ultra-violet (UV) laser diode coupled to an optical fibre for the local activation of caged compounds combined with a Micro-Electrode Array (MEA) device. Local uncaging was achieved by UV irradiation through the optical fibre previously positioned by using a red laser diode. The size of the stimulation was determined using caged fluorescein, whereas its efficacy was tested by studying the effect of uncaging the neurotransmitter glutamate. Uncaged glutamate evoked neuronal responses that were recorded using either fluorescence measurements or electrophysiological recordings with MEAs, thus showing the ability of our system to induce local neuronal excitation. This method allows overcoming the limitations of the MEA system related to unfocused electrical stimulation and induction of electrical artefacts. In addition, the coupling of a UV laser diode to an optical fibre allows a precise local stimulation and a quick change of the stimulation point.


The regulatory network of natural competence and transformation of Vibrio cholerae.
The human pathogen Vibrio cholerae is an aquatic bacterium frequently encountered in rivers, lakes, estuaries, and coastal regions. Within these environmental reservoirs, the bacterium is often found associated with zooplankton and more specifically with their chitinous exoskeleton. Upon growth on such chitinous surfaces, V. cholerae initiates a developmental program termed "natural competence for genetic transformation." Natural competence for transformation is a mode of horizontal gene transfer in bacteria and contributes to the maintenance and evolution of bacterial genomes. In this study, we investigated competence gene expression within this organism at the single cell level. We provide evidence that under homogeneous inducing conditions the majority of the cells express competence genes. A more heterogeneous expression pattern was observable on chitin surfaces. We hypothesize that this was the case due to the heterogeneity around the chitin surface, which might vary extensively with respect to chitin degradation products and autoinducers; these molecules contribute to competence induction based on carbon catabolite repression and quorum-sensing pathways, respectively. Therefore, we investigated the contribution of these two signaling pathways to natural competence in detail using natural transformation assays, transcriptional reporter fusions, quantitative RT-PCR, and immunological detection of protein levels using Western blot analysis. The results illustrate that all tested competence genes are dependent on the transformation regulator TfoX. Furthermore, intracellular cAMP levels play a major role in natural transformation. Finally, we demonstrate that only a minority of genes involved in natural transformation are regulated in a quorum-sensing-dependent manner and that these genes determine the fate of the surrounding DNA. We conclude with a model of the regulatory circuit of chitin-induced natural competence in V. cholerae.


Null mutation of the Lmo4 gene or a combined null mutation of the Lmo1/Lmo3 genes causes perinatal lethality, and Lmo4 controls neural tube development in mice.
The LIM-only family of proteins comprises four members; two of these (LMO1 and LMO2) are involved in human T-cell leukemia via chromosomal translocations, and LMO2 is a master regulator of hematopoiesis. We have carried out gene targeting of the other members of the LIM-only family, viz., genes Lmo1, Lmo3 and Lmo4, to investigate their role in mouse development. None of these genes has an obligatory role in lymphopoiesis. In addition, while null mutations of Lmo1 or Lmo3 have no discernible phenotype, null mutation of Lmo4 alone causes perinatal lethality due to a severe neural tube defect which occurs in the form of anencephaly or exencephaly. Since the Lmo1 and Lmo3 gene sequences are highly related and have partly overlapping expression domains, we assessed the effect of compound Lmo1/Lmo3 null mutations. Although no anatomical defects were apparent in compound null pups, these animals also die within 24 h of birth, suggesting that a compensation between the related Lmo1 and 3 proteins can occur during embryogenesis to negate the individual loss of these genes. Our results complete the gene targeting of the LIM-only family in mice and suggest that all four members of this family are important in regulators of distinct developmental pathways.


Enhancing resolution along multiple imaging dimensions using assorted pixels.
Multisampled imaging is a general framework for using pixels on an image detector to simultaneously sample multiple dimensions of imaging (space, time, spectrum, brightness, polarization, etc.). The mosaic of red, green, and blue spectral filters found in most solid-state color cameras is one example of multisampled imaging. We briefly describe how multisampling can be used to explore other dimensions of imaging. Once such an image is captured, smooth reconstructions along the individual dimensions can be obtained using standard interpolation algorithms. Typically, this results in a substantial reduction of resolution (and, hence, image quality). One can extract significantly greater resolution in each dimension by noting that the light fields associated with real scenes have enormous redundancies within them, causing different dimensions to be highly correlated. Hence, multisampled images can be better interpolated using local structural models that are learned offline from a diverse set of training images. The specific type of structural models we use are based on polynomial functions of measured image intensities. They are very effective as well as computationally efficient. We demonstrate the benefits of structural interpolation using three specific applications. These are 1) traditional color imaging with a mosaic of color filters, 2) high dynamic range monochrome imaging using a mosaic of exposure filters, and 3) high dynamic range color imaging using a mosaic of overlapping color and exposure filters.


RNAconTest: comparing tools for noncoding RNA multiple sequence alignment based on structural consistency.
The importance of noncoding RNA sequences has become increasingly clear over the past decade. New RNA families are often detected and analyzed using comparative methods based on multiple sequence alignments. Accordingly, a number of programs have been developed for aligning and deriving secondary structures from sets of RNA sequences. Yet, the best tools for these tasks remain unclear because existing benchmarks contain too few sequences belonging to only a small number of RNA families. RNAconTest (RNA consistency test) is a new benchmarking approach relying on the observation that secondary structure is often conserved across highly divergent RNA sequences from the same family. RNAconTest scores multiple sequence alignments based on the level of consistency among known secondary structures belonging to reference sequences in their output alignment. Similarly, consensus secondary structure predictions are scored according to their agreement with one or more known structures in a family. Comparing the performance of 10 popular alignment programs using RNAconTest revealed that DAFS, DECIPHER, LocARNA, and MAFFT created the most structurally consistent alignments. The best consensus secondary structure predictions were generated by DAFS and LocARNA (via RNAalifold). Many of the methods specific to noncoding RNAs exhibited poor scalability as the number or length of input sequences increased, and several programs displayed substantial declines in score as more sequences were aligned. Overall, RNAconTest provides a means of testing and improving tools for comparative RNA analysis, as well as highlighting the best available approaches. RNAconTest is available from the DECIPHER website (http://DECIPHER.codes/Downloads.html).


Do expanded seven-day NHS services improve clinical outcomes? Analysis of comparative institutional performance from the "NHS Services, Seven Days a Week" project 2013-2016.
The cause of adverse weekend clinical outcomes remains unknown. In 2013, the "NHS Services, Seven Days a Week" project was initiated to improve access to services across the seven-day week. Three years on, we sought to analyse the impact of such changes across the English NHS. Aggregated trust-level data on crude mortality rates, Summary Hospital-Level Mortality Indicator (SHMI), mean length of stay (LOS), A&E admission and four-hour breach rates were obtained from national Hospital Episode Statistics and A&E datasets across the English NHS, excluding mental and community health trusts. Trust annual reports were analysed to determine the presence of any seven-day service reorganisation in 2013-2014. Funnel plots were generated to compare institutional performance and a difference in differences analysis was performed to determine the impact of seven-day changes on clinical outcomes between 2013 and 2014, 2014-2015 and 2015-2016. Data was summarised as mean (SD). Of 159 NHS trusts, 79 (49.7%) instituted seven-day changes in 2013-2014. Crude mortality rates, A&E admission rates and mean LOS remained relatively stable between 2013 and 2016, whilst A&E four-hour breach rates nearly doubled from 5.3 to 9.7%. From 2013 to 2014 to 2014-2015 and 2015-2016, there were no significant differences in the change in crude mortality (2014-2015 p = 0.8, 2015-2016 p = 0.9), SHMI (2014-2015 p = 0.5, 2015-2016 p = 0.5), mean LOS (2014-2015 p = 0.5, 2015-2016 p = 0.4), A&E admission (2014-2015 p = 0.6, 2015-2016 p = 1.0) or four-hour breach rates (2014-2015 p = 0.06, 2015-2016 p = 0.6) between trusts that had implemented seven-day changes compared to those which had not. Adverse weekend clinical outcomes may not be ameliorated by large scale reorganisations aimed at improving access to health services across the week. Such changes may negatively impact care quality without additional financial investment, as demonstrated by worsening of some outcomes. Detailed prospective research is required to determine whether such reallocation of finite resources is clinically effective.


Exercise responses following heart transplantation: 5 year follow-up.
Heart transplantation is an established treatment for end stage heart failure. In addition to increased life expectancy, heart transplant recipients report a remarkable improvement in symptoms and functional capacity. Exercise performance following heart transplantation, however, remains impaired even in the absence of exertional symptoms. We have assessed the response to exercise in 47 patients with cardiac failure prior to and then at yearly intervals to five years post transplantation. All patients performed incremental symptom limited exercise tests during which minute ventilation (V'E), oxygen consumption (V'O2) and carbon dioxide production (V'CO2) and heart rate (HR) were measured. Ventilatory response (V'E/V'CO2), anaerobic threshold (V'O2 AT %predicted) and heart rate response (HR/VO2) were calculated. The dead space to tidal volume ratio (VD/VT) and alveolar-arterial oxygen gradient (A-aO2) were computed from transcutaneous monitoring. Despite substantial improvement in subjective functional capacity, heart transplant recipients continue to have limited exercise performance [Maximal V'O2% predicted pre-transplant 41.3 (2.2); 1 year 48.6 (1.7), p <0.001: V'O2 AT% 31.5 (1.1); 1 year 35.6 (1.0); respectively p<0.05]. The maximal oxygen uptake continued to improve at two years post-transplant but, thereafter, there was no further significant change at up to 5 years post transplant [50.9 (1.5)]. At one year post-transplantation peak HR [65.2 (0.9) vs 79.1(1.4)] and the HR/VO2 response [24.0(1.8) vs 79.6(4.2)] were significantly reduced compared to pre-transplant values. The heart rate response remained lower compared to predicted at 5 years post-transplant although there was a significant increase compared to one year post-transplant (32.9 vs 24.0mls/bt). There was a weak but significant relationship between maximal VO2 and peak HR (0.39, p<0.05) and HR/VO2 (r= 0.37, p<0.05) at one year post-transplant. Prior to transplantation the ventilatory response to exercise was elevated [V'E/V'CO2 45.6 (2.5)] and decreased significantly following transplantation [1 yr 34.1 (1.3), respectively p<0.001]. In addition, despite significant improvement in VD/VT after transplantation, it remained higher than normal [Pre VD/VT at maximum exercise 0.35 (0.02); 1 yr 0.31 (0.02); p<0.05]. There was a further fall in the VE/VCO2 and VD/VT at two years post-transplantation with no further change at up to 5 years post transplantation [VE/VCO2 32.0 (1.0); VD/VT 0.29 (0.01)]. Although cardiac output is markedly improved after transplantation, due to chronotropic incompetence associated with denervation, its response remains subnormal and this may explain the residual abnormalities of ventilatory and gas exchange responses to exercise following transplantation.


VACTERL-H associated with central hypothyroidism: a case report.
The VACTERL-H syndrome is a rare combination of vertebral anomalies, anal atresia, congenital heart defects, tracheo-esophageal fistula, abnormalities of kidneys and limb anomalies together with hydrocephalus. This condition is recognized as a hereditary entity with poor prognosis. We present a newborn weighing 3400 g, born by cesarean section to a 27 years old mother who had had an irregular antenatal follow-up. The patient had severe hydrocephalus, proximal esophageal atresia and distal tracheoesophageal fistula, gastric outlet obstruction, imperforated anus and recto-urethral fistula, patent ductus arterious, a bifid scrotum, a vertebral defect, sacral dimple and central hypothyroidism. The patient had no limb defects. The association of central hypothyroidism and VACTERL-H has previously not been reported.


Psychiatric phenomena in Alzheimer's disease. IV: Disorders of behaviour.
Out of a sample of 178 patients with AD, aggression was present in 20%, wandering in 19%, binge-eating in 10%, hyperorality in 6%, urinary incontinence in 48%, and sexual disinhibition in 7%. Behavioural abnormalities were greater in those with more severe dementia. Temporallobe atrophy correlated with aggression, and widening of the third ventricle with hyperorality. Features of the Kluver-Bucy syndrome were commonly seen, but the full syndrome occurred in only one subject. Patients with at least one feature of the Kluver-Bucy syndrome had greater temporal-lobe atropy than those without any of the features.


Epithelial stem cells in vivo.
Cellular topography within the highly polarized surface epithelia can be used to identify the location of the stem cells. In some instances, this can be quite precise and allows the characteristics of stem cells to be studied. Our current knowledge of the stem cell population in murine epidermis and small intestinal crypts is reviewed. In the epidermis, the stem cells would appear to make up about 10% of the basal layer and are distributed towards the centre of the basal layer component of the epidermal proliferative unit. These cells have a long cell cycle and are probably the same cells that retain both tritiated thymidine and radioactively labelled carcinogens for long periods of time. This label retention permits the labelling of the putative stem cell compartment. Over recent years, there has been an accumulation of information indicating various types of heterogeneity within the basal layer, much of which can be interpreted in relation to cellular hierarchies. In the small intestine, cell positions can be fairly precisely identified and the stem cell zone identified. Complex modelling of a wide range of cell kinetic experiments suggests that each crypt contains between 4 and 16 steady state functional stem cells. Radiobiological experiments suggest that up to 32 cells may be capable of clonal regeneration. The repopulation of the clonogenic cell compartment has been determined and the doubling time measured to be 19.7 h. Such studies should throw further light on the behaviour of stem cells and identify the timing of periods of increased and decreased cell proliferation (activation and suppression of controls).


The ALS-associated proteins FUS and TDP-43 function together to affect Drosophila locomotion and life span.
The fatal adult motor neuron disease amyotrophic lateral sclerosis (ALS) shares some clinical and pathological overlap with frontotemporal dementia (FTD), an early-onset neurodegenerative disorder. The RNA/DNA-binding proteins fused in sarcoma (FUS; also known as TLS) and TAR DNA binding protein-43 (TDP-43) have recently been shown to be genetically and pathologically associated with familial forms of ALS and FTD. It is currently unknown whether perturbation of these proteins results in disease through mechanisms that are independent of normal protein function or via the pathophysiological disruption of molecular processes in which they are both critical. Here, we report that Drosophila mutants in which the homolog of FUS is disrupted exhibit decreased adult viability, diminished locomotor speed, and reduced life span compared with controls. These phenotypes were fully rescued by wild-type human FUS, but not ALS-associated mutant FUS proteins. A mutant of the Drosophila homolog of TDP-43 had similar, but more severe, deficits. Through cross-rescue analysis, we demonstrated that FUS acted together with and downstream of TDP-43 in a common genetic pathway in neurons. Furthermore, we found that these proteins associated with each other in an RNA-dependent complex. Our results establish that FUS and TDP-43 function together in vivo and suggest that molecular pathways requiring the combined activities of both of these proteins may be disrupted in ALS and FTD.


Immunocytochemical detection of tumor cells in bone marrow and peripheral blood stem cell collections from patients with ovarian cancer.
High-dose chemotherapy (HDC) followed by autologous hematopoietic reconstitution is an experimental treatment option for patients with epithelial ovarian cancer. However, the incidence of occult ovarian tumor cell involvement in autologous bone marrow (BM) or peripheral blood stem cell (PBSC) autografts has not been widely investigated. We used a highly sensitive immunocytochemical (ICC) procedure that detects occult blood-borne tumor micrometastases. We analyzed 24 BM specimens (15 obtained during therapy and 9 harvest samples) and seven PBSC specimens from 22 patients with ovarian cancer. Overall, ICC analysis detected immunostained tumor cells in 10 of 23 evaluable BM specimens (43%) from 9 of 19 patients (47%). One of 9 (11%) harvest samples contained tumor cells. Only one of the 10 ICC-positive BM specimens had tumor cells detected by routine histopathological analysis. ICC-detectable tumor cells were cleared from the marrow of two patients during chemotherapy. None of the seven PBSC specimens contained tumor cells. We conclude that ovarian cancer micrometastases have the potential to contaminate BM, as is also the case in patients with other epithelial malignancies. In the limited number of specimens analyzed, PBSC harvests appeared to provide a less tumor-contaminated source of hematopoietic stem cells for autologous transplantation.


Entropy based fingerprint for local crystalline order.
We introduce a new fingerprint that allows distinguishing between liquid-like and solid-like atomic environments. This fingerprint is based on an approximate expression for the entropy projected on individual atoms. When combined with local enthalpy, this fingerprint acquires an even finer resolution and it is capable of discriminating between different crystal structures.


Women's reproductive rights in the Amazon basin of Ecuador: challenges for transforming policy into practice.
Despite advances made by Ecuador in developing policies on reproductive and sexual rights, implementation, and oversight remain a challenge, affecting in particular those living in the Amazon basin. This paper reports on an evaluation of sexual and reproductive health and rights (SRHR) in Orellana, Ecuador, the basis of which was the Health Rights of Women Assessment Instrument, which was altered to focus on government obligations, the reality of access and utilization of services, and the inequities and implementation challenges between the two. A community-based cross-sectional survey conducted in 2006 served to document the current status of SRHR Local female field workers interviewed 2025 women on three areas of womens reproductive health: delivery care, family planning, and pregnancy among adolescent girls age 10-19. The results suggest a reality more dismal than that of the official information for the area. Skilled delivery care, modern contraceptive use, and wanted pregnancies were conspicuously lower among indigenous women living in rural areas. Access to reproductive health services varied between rural and urban women. These significant differences in care--amongst others documented--raise concerns over the utility of national-level data for addressing inequities. The gaps evident in the validity of available information for monitoring policies and programs, and between national policy and action reveal that much still needs to be done to realize SRHR for women in the Amazon basin, and that current accountability mechanisms are inadequate.


Adult bile duct strictures: role of MR imaging and MR cholangiopancreatography in characterization.
Bile duct strictures in adults are secondary to a wide spectrum of benign and malignant pathologic conditions. Benign causes of bile duct strictures include iatrogenic causes, acute or chronic pancreatitis, choledocholithiasis, primary sclerosing cholangitis, IgG4-related sclerosing cholangitis, liver transplantation, recurrent pyogenic cholangitis, Mirizzi syndrome, acquired immunodeficiency syndrome cholangiopathy, and sphincter of Oddi dysfunction. Malignant causes include cholangiocarcinoma, pancreatic adenocarcinoma, and periampullary carcinomas. Rare causes include biliary inflammatory pseudotumor, gallbladder carcinoma, hepatocellular carcinoma, metastases to bile ducts, and extrinsic bile duct compression secondary to periportal or peripancreatic lymphadenopathy. Contrast material-enhanced magnetic resonance (MR) imaging with MR cholangiopancreatography is extremely helpful in the noninvasive evaluation of patients with obstructive jaundice, an obstructive pattern of liver function, or incidentally detected biliary duct dilatation. Some of these conditions may show characteristic findings at MR imaging-MR cholangiopancreatography that help in making a definitive diagnosis. Although endoscopic retrograde cholangiopancreatography with tissue biopsy or surgery is needed for the definitive diagnosis of many of these strictures, certain MR imaging characteristics of the narrowed segment (eg, thickened wall, long-segment involvement, asymmetry, indistinct outer margin, luminal irregularity, hyperenhancement relative to the liver parenchyma) may favor a malignant cause. Awareness of the various causes of bile duct strictures in adults and familiarity with their appearances at MR imaging-MR cholangiopancreatography are important for accurate diagnosis and optimal patient management.


Geriatric Assessment in Elderly Patients with End-Stage Kidney Disease.
Decision-making in elderly patients considering dialysis is highly complex. With the increasing number of elderly with end-stage kidney disease (ESKD), it may be important to assess geriatric impairments in this population. The aim of the Geriatric assessment in OLder patients starting Dialysis (GOLD) study was to assess the prevalence of geriatric impairments and frailty in the elderly ESKD population by means of a geriatric assessment (GA), which is a comprehensive tool for overall health assessment. This study included 285 patients ≥65 years: 196 patients at the time of dialysis initiation and 89 patients who chose maximal conservative management (MCM). The GA assessed cognition, mood, nutritional status, (instrumental) activities of daily living (ADL), mobility, comorbidity burden, quality of life and overall frailty. The mean age of the participants was 78 years and 36% were women. Of the incident dialysis patients, 77% started haemodialysis and 23% started peritoneal dialysis. Geriatric impairments were highly prevalent in both dialysis and MCM patients. Most frequently impaired geriatric domains in the dialysis group were functional performance (ADL 29%, instrumental ADL (iADL) 79%), cognition (67%) and comorbidity (41%). According to the GA, 77% in the dialysis group and 88% in the MCM group had 2 or more geriatric impairments. In the MCM group, functional impairment (ADL 45%, iADL 85%) was highly prevalent. Geriatric impairments are highly prevalent in the elderly ESKD population. Since impairments can be missed when not searched for in regular (pre)dialysis care, the first step of improving nephrologic care is awareness of the extensiveness of geriatric impairment.


Evaluation of a training curriculum for prehospital trauma ultrasound.
In the United States, ultrasound has rarely been incorporated into prehospital care, and scant descriptions of the processes used to train prehospital providers are available. Our objective was to evaluate the effectiveness of an extended focused assessment with sonography for trauma (EFAST) training curriculum that incorporated multiple educational modalities. We also aimed to determine if certain demographic factors predicted successful completion. All aeromedical prehospital providers (APPs) for a Level I trauma center took a 25-question computer-based test to ascertain baseline knowledge. Questions were categorized by content and format. Training over a 2-month period included a didactic course, a hands-on training session, proctored scanning sessions in the Emergency Department, six Internet-based training modules, pocket flashcards, a review session, and remedial training. At the conclusion of the training curriculum, the same test and an objective structured clinical examination were administered to evaluate knowledge gained. Thirty-three of 34 APPs completed training. The overall pre-test and post-test means and all content and format subsets showed significant improvement (p < 0.0001 for all). No APP passed the pre-test, and 28 of 33 passed the post-test with a mean score of 78%. No demographic variable predicted passing the post-test. Twenty-seven of 33 APPs passed the objective structured clinical examination, and the only predictive variable was passing the post-test (odds ratio 1.21, 95% confidence interval 1.00-1.25, p = 0.045). The implementation of a multifaceted EFAST prehospital training program is feasible. Significant improvement in overall and subset testing scores suggests that the test instrument was internally consistent and sufficiently sensitive to capture knowledge gained as a result of the training. Demographic variables were not predictive of test success.


Imagining real and unreal things: evidence of a dissociation in autism.
Current theories of visual imagery hold that the same neural processes govern both the representation of real objects and the representation of imagined (but real) objects. Here we test whether the representation of imagined (real) objects and the representation of imagined (but unreal) objects depend on the same or different neurocognitive processes. A likely clinical group for a dissociation between these two types of imagination are children with autism, since they show deficits in imaginative play, impoverished imagination is part of their diagnosis, but they can search for hidden objects. The present study explored imagination in autism using experimental methods. Experiment 1 investigated if children with autism could introduce changes to their representations of people and houses, using Karmiloff-Smith's (1989) technique of asking children to draw "impossible" people or houses. Results showed that children with autism were significantly worse than matched controls in their ability to introduce "unreal" changes to their representations of people and houses. Instead, they tended to draw real people or objects. Experiment 2 investigated whether the performance in Experiment 1 by children with autism was due to an inability to disengage from "real world" representations, as executive dysfunction theorists would argue. To do this, the experimenter instructed them on what to draw and how to draw it. Results showed that even when executive control passed to the experimenter in this way, the children with autism were still significantly impaired in their ability to draw imaginary but unreal things relative to the matched controls. Experiment 3 investigated whether the results from Experiments 1 and 2 arose because of a generativity deficit in autism, which might be the executive dysfunction theorists' alternative account. To test this, the same subjects were given a test of Verbal Fluency and a test of imagining multiple functions of a brick. Results showed that the children with autism were no worse than clinical controls in their ability to generate ideas about real objects, suggesting that a global generativity deficit cannot explain the previous findings. Rather, these results point to a specific impairment in the ability to imagine unreal objects. This is discussed in terms of its possible neural dissociability from other kinds of imagery, and in terms of its possible relationship to theory of mind.


Grasping trapezoidal objects.
When grasping rectangular or circular objects with a precision grip the digits close in on the object in opposite directions. In doing so the digits move perpendicular to the local surface orientation as they approach opposite sides of the object. This perpendicular approach is advantageous for accurately placing the digits. Trapezoidal objects have non-parallel surfaces so that moving the digits in opposite directions would make the digits approach the contact surfaces at an angle that is not 90 degrees . In this study we examined whether this happens, or whether subjects tend to approach trapezoidal objects' surfaces perpendicularly. We used objects of different sizes and with different surface slants. Subjects tended to approach the object's surfaces orthogonally, suggesting that they aim for an optimal precision of digit placement rather than simply closing their hand as it reaches the object.


Explaining the success of Kogelnik's coupled-wave theory by means of perturbation analysis: discussion.
The problem of diffraction of an electromagnetic wave by a thick hologram grating can be solved by the famous Kogelnik's coupled-wave theory (CWT) to a very high degree of accuracy. We confirm this finding by comparing the CWT and the exact result for a typical example and propose an explanation in terms of perturbation theory. To this end we formulate the problem of diffraction as a matrix problem following similar well-known approaches, especially rigorous coupled-wave theory (RCWT). We allow for a complex permittivity modulation and a possible phase shift between refractive index and absorption grating and explicitly incorporate appropriate boundary conditions. The problem is solved numerically exact for the specific case of a planar unslanted grating and a set of realistic values of the material's parameters and experimental conditions. Analogously, the same problem is solved for a two-dimensional truncation of the underlying matrix that would correspond to a CWT approximation but without the usual further approximations. We verify a close coincidence of both results even in the off-Bragg region and explain this result by means of a perturbation analysis of the underlying matrix problem. Moreover, the CWT is found not only to coincide with the perturbational approximation in the in-Bragg and the extreme off-Bragg cases, but also to interpolate between these extremal regimes.


By the pricking of my thumbs, something wicked this way comes: sporadic cancers versus eponymous hereditary cancer predisposition syndromes.
Advances in cancer genomics have led to the recognition of a growing number of high-penetrance single-gene cancer predisposition syndromes. Frequently, the suspicion for a hereditary syndrome is raised by a strongly positive family history. However, other features, such as younger-than-usual age at diagnosis and rare histology should also prompt consideration of a genetic syndrome. Common malignancies frequently show a positive family history without an eponymous syndrome being recognized. This article reports on a case with an unusual constellation of malignancies with distinctive pathologies, which raised suspicion for an eponymous cancer pre-disposition syndrome. Absent a positive family history, a de novo mutation-an alteration in a gene that is present for the first time in a family member as a result of a mutation in a germ cell of one of the parents or in the fertilized ovum-was suspected. The authors discuss indications for genetic counseling and testing, limitations, and the evidence that supports the recommendations as formulated by working groups and the NCCN. Most frequently, these recommendations are reasonable statements based on the natural history of the disease, but without population-based studies for many rare syndromes, the actual penetrance, variable expressivity, and actual associated cancer risk are unknown.


South African guideline for the diagnosis, management and prevention of acute viral bronchiolitis in children.
South African Thoracic Society, South African Society of Paediatric Infectious Diseases, United South African Neonatal Association. To develop and publish a guideline for doctors managing acute viral bronchiolitis, because this condition is extremely common in South Africa, it is responsible for significant morbidity in the population, and subsequently a great deal of patient and parental distress, and the disease is costly, since many children are unnecessarily subjected to investigations and treatment strategies that are of no proven benefit. The main aims of the guideline are to promote an improved standard of treatment based on understanding of the disease and its management, and to encourage cost-effective and appropriate management. A detailed literature review was conducted and summarised into this document by a selected working group of paediatricians from around the country. Recommendations. These include the appropriate diagnostic and management strategies for acute viral bronchiolitis.


Humanized OKT3 antibodies: successful transfer of immune modulating properties and idiotype expression.
Antibodies that possess the Ag-binding regions of OKT3 within the context of a human framework (Hu-OKT3 Ab) offer distinct advantages for optimizing anti-CD3 mAb therapy. First, manipulation of Ab genes to produce humanized Ab that retain Ag-binding activity may circumvent antigenicity problems. Second, Ab gene engineering provides a means for modifying functional properties, including T cell activation and immune suppression. The purpose of this study was to determine the functional properties of Hu-OKT3 Ab and to compare the functional properties and idiotypes of Hu-OKT3 Ab to those of murine OKT3. Three Hu-OKT3 IgG4 Ab, a chimeric OKT3 antibody (cOKT3-1) (grafted sequences comprising all OKT3 VH and VL regions) and two complementarity determining region (CDR)-grafted antibodies, gOKT3-5 and gOKT3-6 (grafted sequences comprising only OKT3 VH and VL CDR and some framework amino acids, were analyzed. Initial studies demonstrated that the cOKT3 and gOKT3-5 Ab bound selectively to T cells and competitively inhibited OKT3-FITC binding with avidities similar to that of murine OKT3. Binding avidity of the gOKT3-6 Ab was markedly less than that of the other two Hu-OKT3 Ab. Serologic analysis suggested that cOKT3 and gOKT3-5 Ab possess idiotypes (combining sites) similar to murine OKT3. T cell activation potency of all three Hu-OKT3 Ab was assessed by proliferation, induction of activation marker expression (IL-2R and Leu 23), and lymphokine production (TNF-alpha and IFN-gamma). The cOKT3 and gOKT3-5 Ab demonstrated T cell activation potencies similar to murine OKT3 as assessed by each parameter. CD3 coating and modulation by these two Ab was effective but somewhat less potent than that observed with OKT3. Finally, cOKT3 and gOKT3-5 Ab both inhibited CTL activity comparably to murine OKT3. In conclusion, these studies indicate that gOKT3-5 and cOKT3 Ab possess immune modulating properties similar to murine OKT3 and thus offer attractive alternatives to murine OKT3 for in vivo therapy.


The binocular organization of simple cells in the cat's visual cortex.
We have studied the manner by which inputs from the two eyes are combined in simple cells of the cat's visual cortex. The stimuli for this study are drifting sinusoidal gratings, shown dichoptically at optimal spatial frequency and orientation. The relative spatial phase (disparity) between the gratings for left and right eyes is varied over 360 degrees. Most simple cells show phase-specific binocular interaction such that response amplitudes and phases vary depending on the relative spatial phase. At one phase, response is greater than either of the monocular responses and often greater than the sum of the two. At the phase 180 degrees away from the optimal, the cell's responses are strongly inhibited and often completely suppressed. Phase-specific binocular interaction disappears when the gratings presented to one eye are made orthogonal to the optimal orientation. The degree of binocular interaction does not depend critically on the ocular dominance of the cells. Simple cells that are nearly equally dominated by each eye always exhibit strong phase-specific interaction. The majority of cells that are strongly dominated by one eye, and even those that appear monocular, show phase-dependent changes in responses. We examined the extent of binocular interaction for cells with preferred orientations near vertical compared with those tuned to other optimal orientations. If these cells are conveying information about depth, one might expect a greater degree of binocular phase-specificity for units preferring nearly vertical orientations, which would then be processing horizontal disparities. We find no evidence for this. Predictions of simple-cell responses are derived from a linear model of binocular convergence in which light-evoked neural signals from each eye are summed linearly to determine cell responses. Data from cells generally follow the prediction of the model for both response amplitude and phase. Deviations from predictions of the linear model are found for a minority of cells. This deviation may be accounted for by a threshold mechanism that comes into play after the linear binocular summation. A small proportion of simple cells that appear monocular by alternate tests of each eye show a purely inhibitory influence from the silent eye. This inhibition is not generally dependent on the relative phase of the gratings. We conclude that most binocular interaction in striate simple cells may be accounted for by linear summation of neural signals from each eye.(ABSTRACT TRUNCATED AT 400 WORDS)


[Prion-like Properties of Misfolded Cu/Zn-superoxide Dismutase in Amyotrophic Lateral Sclerosis: Update and Perspectives].
Amyotrophic lateral sclerosis (ALS) is a lethal neurodegenerative disease that is characterized by the loss of motor neurons, which results in progressive muscle atrophy. The pathology spreads from the initial site of onset to contiguous anatomic regions. Mutations in the gene encoding Cu/Zn-superoxide dismutase (SOD1) have been identified in a dominantly inherited form of ALS (ALS-SOD1). A major hallmark of ALS-SOD1 is the abnormal accumulation of conformationally aberrant SOD1 protein (i.e., misfolded SOD1) within motor neurons. Emerging experimental evidence has suggested that misfolded proteins associated with neurodegenerative diseases exhibit prion-like properties, i.e., misfolded proteins act as conformational templates that convert normal proteins into a pathogenic form. Possibly as a result of this prion-like self-propagation property, misfolded forms of pathological proteins are considered to accumulate in the central nervous system and cause neurodegeneration. In this article, we review recent evidence for the role of prion-like mechanisms in ALS-SOD1. In particular, we discuss the propensity of misfolded SOD1 to act as a pathological seed, spread between cells, and propagate neuroanatomically.


Does 30-day readmission affect long-term outcome among glioblastoma patients?
Research on readmissions has focused mainly on the economic and resource burden it places on hospitals. To evaluate the effect of 30-day readmission on overall survival among newly diagnosed glioblastoma multiforme (GBM) patients. A nationwide cohort of GBM patients diagnosed between 1991 and 2007 was studied using the Surveillance, Epidemiology and End Results Medicare database. Multivariate models were used to determine factors associated with readmission and overall survival. Odds ratio, hazard ratio, 95% confidence interval, and P values were reported. Complete case and multiple imputation analyses were performed. Among the 2774 newly diagnosed GBM patients undergoing surgery at 442 hospitals nationwide, 437 (15.8%) were readmitted within 30 days of the index hospitalization. Although 63% of readmitted patients returned to the index hospital where surgery was performed, a significant portion (37%) were readmitted to nonindex hospitals. The median overall survival for readmitted patients (6.0 months) was significantly shorter than for nonreadmitted (7.6 months; P < .001). In a confounder-adjusted imputed model, 30-day readmission increased the hazard of mortality by 30% (hazard ratio, 1.3; P < .001). Neurological symptoms (30.2%), thromboembolic complications (19.7%), and infections (17.6%) were the leading reasons for readmission. Prior studies that have reported only the readmissions back to index hospitals are likely underestimating the true 30-day readmission rate. GBM patients who were readmitted within 30 days had significantly shorter survival than nonreadmitted patients. Future studies that attempt to decrease readmissions and evaluate the impact of reducing readmissions on patient outcomes are needed.


Correction of prominent zygomata by tripod osteotomy of the malar bone.
Reduction of the prominent zygomata is occasionally required in aesthetic surgery in Oriental patients. Two approaches to surgical exposure are commonly used; one is oral and the other is coronal. In addition, there are two basic surgical concepts of reduction malar-bone plasty; one is thinning and chiseling the prominent area, and the other is mobilizing the prominent zygoma by osteotomy of the zygomata. We devised a new tripod osteotomy of the zygomata by the coronal exposure approach and applied it to correct prominent zygomata in association with frontoperiorbital lifting in Oriental patients. Satisfactory results were obtained and we concluded that this was an effective procedure to correct prominent zygomata for elderly Oriental patients.


Heart disease mortality following widowhood: some results from the OPCS Longitudinal Study. Office of Population Censuses and Surveys.
Many studies have suggested that following the experience of 'stressful' life events the risks of myocardial infarction, accidents and perhaps other diseases are elevated. In the OPCS Longitudinal Study routinely collected data on deaths, and deaths of a spouse occurring in a 1% sample of the population of England and Wales in the period 1971-1981 are linked together, and with 1971 census records of sample members. The timing and patterns of death following the potentially very stressful event of conjugal bereavement may thus be analysed. Overall the mortality (from ischaemic heart disease) was less than 10% in excess of that in all members of the LS sample. As in many earlier studies, some increases in death rates shortly after widowhood are observed. Unusually, for deaths from all causes these increases are more marked in widows than in widowers with, for example, a two-fold increase in mortality from all causes in the first month after widowhood. However, no peak of post-bereavement mortality from ischaemic heart disease is clearly established in either sex. Although the study is large, with a well-chosen control group, only a limited characterisation of study members from data collected in the census is possible. In particular, no measures of personality, behaviour or diet are available. Investigation of potential effects of social or familial support, as measured by household structure and numbers of children, led to equivocal results. Several possible explanations for the increased mortality rates are examined. Hypotheses based on common marital environment, homogamy or simultaneous accidental death are seen to be of very limited value. The observed patterns, although consistent with an early effect of a stressful life event, do not suggest that stress following bereavement leads to an excess of ischaemic heart disease mortality.


Incorporation of caseinoglycomacropeptide and caseinophosphopeptide into the salivary pellicle inhibits adherence of mutans streptococci.
The protective effects of milk and milk products against dental caries have been demonstrated in many animal studies. We have shown that this effect was mediated by micellar casein or caseinopeptide derivatives. A reduction in the Streptococcus sobrinus population in the oral microbiota of animals fed diets supplemented with these milk components was consistently observed. A possible explanation for these findings is that milk components are incorporated into the salivary pellicle, thereby reducing the adherence of S. sobrinus. This hypothesis was tested in vitro by the incubation of bovine enamel discs with unstimulated saliva. The resulting pellicle was washed and incubated with caseinoglycomacropeptide (CGMP) and/or caseinophosphopeptide (CPP) labeled with 17- and 12-nm gold particles. All samples were prepared for electron microscopy by high-pressure freezing followed by freeze-substitution. It was demonstrated by high-resolution scanning electron microscopy with back-scattered electron imaging, as well as by transmission electron microscopy, that both peptides were incorporated into the pellicle in exchange for albumin, confirming previous findings. This protein was identified with a mouse anti-human serum albumin followed by goat anti-mouse IgG labeled with 25-nm gold particles. Incorporation of CGMP and/or CPP into salivary pellicles reduced the adherence of both S. sobrinus and S. mutans significantly. It is suggested that the calcium and phosphate-rich micellar casein or caseinopeptides are incorporated into the pellicle. The resulting ecological shifts, together with the increased remineralization potential of this biofilm, may explain its modified cariogenic potential.


A Virtual Emergency Telemedicine Serious Game in Medical Training: A Quantitative, Professional Feedback-Informed Evaluation Study.
Serious games involving virtual patients in medical education can provide a controlled setting within which players can learn in an engaging way, while avoiding the risks associated with real patients. Moreover, serious games align with medical students' preferred learning styles. The Virtual Emergency TeleMedicine (VETM) game is a simulation-based game that was developed in collaboration with the mEducator Best Practice network in response to calls to integrate serious games in medical education and training. The VETM game makes use of data from an electrocardiogram to train practicing doctors, nurses, or medical students for problem-solving in real-life clinical scenarios through a telemedicine system and virtual patients. The study responds to two gaps: the limited number of games in emergency cardiology and the lack of evaluations by professionals. The objective of this study is a quantitative, professional feedback-informed evaluation of one scenario of VETM, involving cardiovascular complications. The study has the following research question: "What are professionals' perceptions of the potential of the Virtual Emergency Telemedicine game for training people involved in the assessment and management of emergency cases?" The evaluation of the VETM game was conducted with 90 professional ambulance crew nursing personnel specializing in the assessment and management of emergency cases. After collaboratively trying out one VETM scenario, participants individually completed an evaluation of the game (36 questions on a 5-point Likert scale) and provided written and verbal comments. The instrument assessed six dimensions of the game: (1) user interface, (2) difficulty level, (3) feedback, (4) educational value, (5) user engagement, and (6) terminology. Data sources of the study were 90 questionnaires, including written comments from 51 participants, 24 interviews with 55 participants, and 379 log files of their interaction with the game. Overall, the results were positive in all dimensions of the game that were assessed as means ranged from 3.2 to 3.99 out of 5, with user engagement receiving the highest score (mean 3.99, SD 0.87). Users' perceived difficulty level received the lowest score (mean 3.20, SD 0.65), a finding which agrees with the analysis of log files that showed a rather low success rate (20.6%). Even though professionals saw the educational value and usefulness of the tool for pre-hospital emergency training (mean 3.83, SD 1.05), they identified confusing features and provided input for improving them. Overall, the results of the professional feedback-informed evaluation of the game provide a strong indication of its potential as an educational tool for emergency training. Professionals' input will serve to improve the game. Further research will aim to validate VETM, in a randomized pre-test, post-test control group study to examine possible learning gains in participants' problem-solving skills in treating a patient's symptoms in an emergency situation.


Relationship between mean platelet volume, platelet distribution width, plateletcrit and varicocele.
Increases in platelet count and platelet indices such as mean platelet volume (MPV), platelet distribution width (RDW) and plateletcrit (PCT) have been reported in a wide range of pathological settings. The aetiology of varicocele, a vascular disease, is poorly defined. In this study, we aimed to examine the relationship between varicocele, platelet count and a series of platelet indices. A total of 69 patients with varicocele and 56 patients without varicocele were enrolled in the study. Patient sperm parameters, platelet, MPV, PDW and PCT values were analysed. There were semen abnormalities in 37 (53.6%) patients in the varicocele group and 19 (33.9%) patients in nonvaricocele group (P < 0.05). There was no significant difference in platelet count or platelet indices (MPV, PDW and PCT) between the patients with and without varicocele (P < 0.05). There was no statistically significant difference in platelet count or MPV, PDW and PCT between patients with varicocele and the control subjects.


Drosophila Ana1 is required for centrosome assembly and centriole elongation.
Centrioles organise centrosomes and cilia, and these organelles have an important role in many cell processes. In flies, the centriole protein Ana1 is required for the assembly of functional centrosomes and cilia. It has recently been shown that Cep135 (also known as Bld10) initially recruits Ana1 to newly formed centrioles, and that Ana1 then recruits Asl (known as Cep152 in mammals) to promote the conversion of these centrioles into centrosomes. Here, we show that ana1 mutants lack detectable centrosomes in vivo, that Ana1 is irreversibly incorporated into centrioles during their assembly and appears to play a more important role in maintaining Asl at centrioles than in initially recruiting Asl to centrioles. Unexpectedly, we also find that Ana1 promotes centriole elongation in a dose-dependent manner: centrioles are shorter when Ana1 dosage is reduced and are longer when Ana1 is overexpressed. This latter function of Ana1 appears to be distinct from its role in centrosome and cilium function, as a GFP-Ana1 fusion lacking the N-terminal 639 amino acids of the protein can support centrosome assembly and cilium function but cannot promote centriole over-elongation when overexpressed.


Stereotactic radiosurgery for metastatic spine tumors.
Spinal metastases invariably affect the majority of patients with cancer. Many will develop symptoms related to pain and disability from epidural spinal cord compression as well as pathologic fracture of the vertebrae. With the emergence of targeted systemic therapies and a better understanding of cancer biology, patients are living longer with bony metastases. This poses particular challenges, as palliation of pain and maintenance of local tumor control are paramount to quality of life and overall functional independence for these patients. Stereotactic radiosurgery (SRS) has emerged as a potent primary standalone and adjuvant treatment option for spinal metastases. To date, the primary indications for SRS include 1) upfront standalone treatment for painful bony metastases in the oligometastatic patient, 2) standalone or post-operative treatment following progression or recurrence of local disease despite previous conventional external beam radiation therapy (cEBRT), and 3) following surgery during which epidural disease is decompressed and the spine stabilized when indicated. SRS has demonstrated a significant advantage over cEBRT for tumors traditionally regarded as relatively radioresistant such as sarcoma, melanoma, renal cell carcinoma, non-small cell lung cancer and colon carcinoma.9 The radiobiological advantage of increased tumoricidal dose delivery and spinal cord dose sparing in SRS have made this a powerful treatment alternative to cEBRT particularly within the context of re-irradiation. Given the limitations of spinal cord dose constraints, surgery is still the first-line therapy in patients with high-grade epidural spinal cord compression (ESCC). Epidural compression can be treated with SRS, however this risks radiation-induced myelopathy and challenges the safety of effective dose delivery at the dural margin.11 With increasing dose, radiation-induced vertebral fracture is the most serious and prevalent side effect of SRS.53 An overview of SRS, including the most common indications, complications, and outcomes for spinal metastases are presented here.


Adjuvant Sandwich Chemotherapy Plus Radiotherapy vs Adjuvant Chemotherapy Alone for Locally Advanced Bladder Cancer After Radical Cystectomy: A Randomized Phase 2 Trial.
Locoregional failure for patients with locally advanced bladder cancer (LABC) after radical cystectomy (RC) is common even with chemotherapy and is associated with high morbidity and mortality. Adjuvant radiotherapy (RT) can decrease locoregional failure but has not been studied in the chemotherapy era. To investigate if adjuvant sequential RT plus chemotherapy can improve locoregional recurrence-free survival (LRFS) compared with adjuvant chemotherapy alone. A randomized phase 3 trial was opened to compare adjuvant RT vs sequential chemotherapy plus RT after RC for LABC, but a third arm was added later as a randomized phase 2 trial to compare chemotherapy plus RT vs adjuvant chemotherapy alone, an emerging standard. The intent-to-treat phase 2 trial reported herein enrolled patients from December 2002 to July 2008. Data were analyzed from August 3, 2015, to January 6, 2016. Routine follow-up and surveillance pelvic computed tomographic (CT) scans every 6 months during the first 2 years were performed. The setting was an academic center. Patients with bladder cancer 70 years or younger having 1 or more risk factors (≥pT3b, grade 3, or positive nodes) with negative margins after radical cystectomy plus pelvic lymph node dissection were eligible. Patients had Eastern Cooperative Oncology Group performance status of 0 to 2, no evidence of distant metastases on CT scan of the abdomen and pelvis or on chest imaging, and adequate renal, hepatic, and hematologic function. Ninety-one percent (109 of 120) had ≥ pT3 disease. Chemotherapy plus RT included 2 cycles of gemcitabine (1000 mg/m2 intravenously on days 1, 8, and 15) and cisplatin (70 mg/m2 intravenously on day 2) before and after RT to 4500 cGy in 150 cGy twice-daily fractions over 3 weeks using 3-dimensional conformal techniques. Chemotherapy alone included 4 cycles of gemcitabine and cisplatin. Locoregional recurrence-free survival. The chemotherapy plus RT arm accrued 75 patients, and the chemotherapy-alone arm accrued 45 patients, with a weighted randomization to speed accrual. Fifty-three percent (64 of 120) had urothelial carcinoma, and 46.7% (56 of 120) had squamous cell carcinoma or other. The arms were balanced except for age (median, 52 vs 55 years; P = .04) and tumor size (mean, 4.9 vs 5.8 cm; P < .01), both favoring chemotherapy plus RT. Two-year outcomes and overall adjusted hazard ratios (HRs) for chemotherapy plus RT vs chemotherapy alone were 96% vs 69% (HR, 0.08; 95% CI, 0.02-0.39; P < .01) for LRFS, 68% vs 56% (HR, 0.53; 95% CI, 0.27-1.06; P = .07) for disease-free survival, and 71% vs 60% (HR, 0.61; 95% CI, 0.33-1.11; P = .11) for overall survival (OS). Five patients (7%) had RT-associated late grade 3 gastrointestinal tract adverse effects in the chemotherapy plus RT arm. Adjuvant chemotherapy plus RT was reasonably well tolerated and was associated with significant improvements in LRFS and marginal improvements in disease-free survival vs chemotherapy alone in LABC. The addition of adjuvant RT should be considered for LABC. This regimen warrants further study in phase 3 trials. clinicaltrials.gov Identifier: NCT01734798.


Cystic gene dosage influences kidney lesions after nephron reduction.
Cystic kidney disease is characterized by the progressive development of multiple fluid-filled cysts. Cysts can be acquired, or they may appear during development or in postnatal life due to specific gene defects and lead to renal failure. The most frequent form of this disease is the inherited polycystic kidney disease (PKD). Experimental models of PKD showed that an increase of cellular proliferation and apoptosis as well as defects in apico-basal and planar cell polarity or cilia play a critical role in cyst development. However, little is known about the mechanisms and the mediators involved in acquired cystic kidney diseases (ACKD). In this study, we used the nephron reduction as a model to study the mechanisms underlying cyst development in ACKD. We found that tubular dilations after nephron reduction recapitulated most of the morphological features of ACKD. The development of tubular dilations was associated with a dramatic increase of cell proliferation. In contrast, the apico-basal polarity and cilia did not seem to be affected. Interestingly, polycystin 1 and fibrocystin were markedly increased and polycystin 2 was decreased in cells lining the dilated tubules, whereas the expression of several other cystic genes did not change. More importantly, Pkd1 haploinsufficiency accelerated the development of tubular dilations after nephron reduction, a phenotype that was associated to a further increase of cell proliferation. These data were relevant to humans ACKD, as cystic genes expression and the rate of cell proliferation were also increased. In conclusion, our study suggests that the nephron reduction can be considered a suitable model to study ACKD and that dosage of genes involved in PKD is also important in ACKD.


The quality of life of home-ventilated children and their primary caregivers plus the associated social and economic burdens: a prospective study.
Despite the obvious challenges faced by families caring for children on home ventilation, there is surprisingly little research into the details of their daily lives. In particular, little is known about the quality of life of the child and caregiver plus the associated social and economic burdens of care. We prospectively studied 90 families enrolled in a paediatric home ventilation service in British Columbia. In the clinic, we recorded demographic information, patient acuity score and quality of life for patient and caregiver using standardised questionnaires. Parents then monitored social and financial costs of care at home over the subsequent 8 weeks. These data were collected by telephone at 1 and 2 months. Most children led rich active lives. Camping trips, wheelchair sports and foreign travel were the norm, not the exception. Over 90% assessed the burden of care as mild or moderate. Government support covers medical expenses and home nursing (median 32 h/week, IQR 0-62.5 h). Monthly unreimbursed family expenses were low (median $87.7, IQR $15.3-$472). Despite this, nearly 25% of primary caregivers assessed burden of care as severe and over 50% had chronic illnesses requiring daily medication (principally depression, anxiety and arthritis). Quality of life for children or caregivers did not correlate with income or education. Home ventilation of complex children is a successful strategy but it places significant strain on the primary caregiver. Specific attention to the physical and mental health of the caregiver should be an integral part of the management of home-ventilated children. NCT01863992.


Through the eye of the beholder: multiple perspectives on quality in women's health care.
Quality is an illusive concept with different meanings to different people. Providers often define quality in terms of patient outcomes, professional standards of practice, predetermined criteria used to measure quality, and even subjective opinion. Patients describe quality in terms of the interpersonal aspects of care, how well they were treated, and the responsiveness of the provider to their needs. This qualitative study using a semi-structured interview defined quality from the perspectives of patients, physicians, nurses, and payers associated with a hospital-based women's service line, and how the attributes of quality varied among the multiple groups. The study also described how stakeholders become aware of quality and how they determined a hospital's quality. From the findings of the study, a conceptual framework of quality in women's health was developed.


Ultrasound-Stimulated Drug Delivery Using Therapeutic Reconstituted High-Density Lipoprotein Nanoparticles.
The abnormal tumor vasculature and the resulting abnormal microenvironment are major barriers to optimal chemotherapeutic drug delivery. It is well known that ultrasound (US) can increase the permeability of the tumor vessel walls and enhance the accumulation of anticancer agents. Reconstituted high-density lipoproteins (rHDL) nanoparticles (NPs) allow selective delivery of anticancer agents to tumor cells via their overexpressed scavenger receptor type B1 (SR-B1) receptor. The goal of this study is to investigate the potential of noninvasive US therapy to further improve delivery and tumor uptake of the payload from rHDL NPs, preloaded with an infrared dye (IR-780), aimed to establish a surrogate chemotherapeutic model with optical localization. Athymic nude mice were implanted orthotopically with one million breast cancer cells (MDA-MB-231/Luc). Three weeks later, animals were divided into seven groups with comparable mean tumor size: control, low, moderate, and high concentration of rHDL NPs alone groups, as well as these three levels of rHDL NPs plus US therapy groups (N = 7 to 12 animals per group), where low, moderate and high denote 5, 10, and 50 µg of the IR-780 dye payload per rHDL NP injection, respectively. The US therapy system included a single element focused transducer connected in series with a function generator and power amplifier. A custom 3D printed cone with an acoustically transparent aperture and filled with degassed water allowed delivery of focused US energy to the tumor tissue. US exposure involved a pulsed sequence applied for a duration of 5 min. Each animal in the US therapy groups received a slow bolus co-injection of MB contrast agent and rHDL NPs. Animals were imaged using a whole-body optical system to quantify intratumoral rHDL NP accumulation at baseline and again at 1 min, 30 min, 24 h, and 48 h. At 48 h, all animals were euthanized and tumors were excised for ex vivo analysis. We investigated a noninvasive optical imaging method for monitoring the effects of US-stimulated drug delivery of IR-780 dye-loaded rHDL NPs in living animals. No change in optical imaging data was found in the control animals. However, there was considerable dye accumulation (surrogate drug) within 48 h in the low (5 µg), moderate (10 µg), and high (50 µg) rHDL NP concentration-dosed group animals (p < 0.09). With US therapy added to the experimental protocol, there was an additional and significant increase in local tumor drug uptake at 48 h (p < 0.02). Optical image data collected from ex vivo tumor samples confirmed tumor retention of the IR-780 dye-loaded rHDL NPs and correlated positively with in vivo optical imaging results (R2 > 0.69, p < 0.003). IR-780 dye extraction from the tumor tissue samples confirmed the in vivo and ex vivo US therapy findings. Overall, the addition of US therapy considerably improved local rHDL NP accumulation in tumor tissue. This study concludes that US-mediated drug delivery can facilitate tumor uptake of rHDL NPs and more research is warranted to optimize the drug dosing schedule and the respective therapeutic protocols.


Family quality of life of Australian families with a member with an intellectual/developmental disability.
Family quality of life (FQOL) is a recent concept in intellectual/developmental disability research. Outcomes for the family are important to the provision of services because families, rather than institutions, are increasingly considered the primary support unit. This article presents Australian findings using the international Family Quality of Life Survey: Main Caregivers of People with Intellectual or Developmental Disabilities (FQOLS-2006). Forty-two South Australian main caregivers of people with an intellectual/developmental disability were interviewed using the FQOLS-2006. The FQOL domains assessed were Health of the Family, Financial Well-being, Family Relationships, Support from Other People, Support from Disability-Related Services, Influence of Values, Careers, Leisure and Recreation, and Community Interaction. Domains were measured in terms of Importance, Opportunities, Attainment, Initiative, Stability and Satisfaction. The FQOLS-2006 asked about the family's practical and emotional Support from Other People together, whereas the current study separated the constructs of practical and emotional support. Questions pertaining to FQOL in the past were also added, in order to gain a broader picture of present FQOL. Results indicated that families considered all the FQOL domains to be important. However, Health, Family Relationships and Financial Well-being were regarded as slightly more important than Practical and Emotional Support from Others. The attainment of Family Relationships, Health, Values, and Leisure and Recreation were rated as quite a bit, but Practical Support from Other People was only rated as a little. Families were generally satisfied with all FQOL domains, but they were satisfied with their Family Relationships and they were neither satisfied or dissatisfied with their Financial Well-being. Results also indicated that there was a need to distinguish between the provision of practical and emotional support from others, because the attainment of emotional support was rated at a slightly higher level than practical support. The FQOLS-2006 provided a comprehensive measure of FQOL, which, with some additional modifications, could be used to better inform service provisions and ultimately enhance the quality of life of people with intellectual/developmental disabilities and their families.


Profiling analysis of long non-coding RNA and mRNA in parathyroid carcinoma.
Parathyroid carcinoma (PCa) is a rare endocrine neoplasia that typically has unfavourable outcomes. The contribution of long non-coding RNAs (lncRNAs) to the development of malignant and benign parathyroid tumours remains largely unknown. In this study, we explored transcriptomic profiling of lncRNA and mRNA expression in 6 PCa, 6 parathyroid adenoma (PAd) and 4 normal parathyroid (PaN) tissues. In total, 2641 lncRNA transcripts and 2165 mRNA transcripts were differentially expressed between PCa and PAd. Enrichment analysis demonstrated that dysregulated transcripts were involved mainly in the extracellular matrix (ECM)-receptor interaction and energy metabolism pathways. Bioinformatics analysis suggested that ATF3, ID1, FOXM1, EZH2 and MITF may be crucial to parathyroid carcinogenesis. Series test of cluster analysis segregated differentially expressed lncRNAs and mRNAs into several expression profile models, among which the 'plateau' profile representing components specific to parathyroid carcinogenesis was selected to build a co-expression network. Seven lncRNAs and three mRNAs were selected for quantitative RT-PCR validation in 16 PCa, 41 PAd and 4 PaN samples. Receiver-operator characteristic curves analysis showed that lncRNA PVT1 and GLIS2-AS1 yielded the area under the curve values of 0.871 and 0.860, respectively. Higher hybridization signals were observed in PCa for PVT1 and PAd for GLIS2-AS1. In conclusion, the current evidence indicates that PAd and PCa partially share common signalling molecules and pathways, but have independent transcriptional events. Differentially expressed lncRNAs and mRNAs have intricate interactions and are involved in parathyroid tumourigenesis. The lncRNA PVT1 and GLIS2-AS1 may be new potential markers for the diagnosis of PCa.


The Impact of the Affordable Care Act's Dependent Coverage Mandate on Use of Dental Treatments and Preventive Services.
Oral health problems are the leading chronic conditions among children and younger adults. Lack of dental coverage is thought to be an important barrier to care but little empirical evidence exists on the causal effect of private dental coverage on use of dental services. We explore the relationship between dental coverage and dental services utilization with an analysis of a natural experiment of increasing private dental coverage stemming from the Affordable Care Act's (ACA)-dependent coverage mandate. To evaluate whether increased private dental insurance due to the spillover effect of the ACA-dependent coverage health insurance mandate affected utilization of dental services among a group of affected young adults. 2006-2013 Medical Expenditure Panel Surveys. We used a difference-in-difference regression approach comparing changes in dental care utilization for 25-year olds affected by the policy to unaffected 27-year olds. We evaluate effects on dental treatments and preventive services RESULTS:: Compared to 27-year olds, 25-year olds were 8 percentage points more likely to have private dental coverage in the 3 years following the mandate. We do not find compelling evidence that young adults increased their use of preventive dental services in response to gaining insurance. We do find a nearly 5 percentage point increase in the likelihood of dental treatments among 25-year olds following the mandate, an effect that appears concentrated among women. Increases in private dental coverage due to the ACA's-dependent coverage mandate do not appear to be driving significant changes in overall preventive dental services utilization but there is evidence of an increase in restorative care.


Adaptive seasonal trend in brood sex ratio: test in two sister species with contrasting breeding systems.
Evolutionary theory predicts adaptive adjustment in offspring sex ratio by females. Seasonal change in sex ratio is one possibility, tested here in two sister species, the Common sandpiper and the Spotted sandpiper Actitis hypoleucos and A. macularia. In the monogamous Common sandpiper, males are the most competitive sex. In each of 3 years, there was a change from mainly sons in early clutches to mainly daughters in late clutches. This seasonal adjustment of clutch sex ratio took place within the female before the eggs were laid, not by differential egg or chick survival. The sex of all eggs laid in the clutches used here was determined molecularly from chick blood taken at the time of hatching. The Spotted sandpiper in contrast is polyandrous, with partly reversed sex roles. There was no seasonal trend from sons to daughters in this species. When tested together, the two species differed significantly as predicted by the hypothesis of adaptive sex ratio adjustment by females.


The effects of low literacy and cognitive impairment on medication adherence in HIV-positive injecting drug users.
Low literacy and cognitive impairment have each been separately identified as risks for non-adherence in HIV infection. However, no studies to date have evaluated these skill deficits in combination. We therefore characterized the nature of literacy and cognitive skills in a sample of HIV-positive injecting drug users and assessed their combined effects on adherence. A community-recruited sample of 57 HIV-positive injecting drug users completed the study. Participants were classified into one of four groups based on their performance on a reading test and a brief neuropsychological battery: high literacy/high cognition, low literacy/high cognition, high literacy/low cognition and low literacy/low cognition. Chi-square and bivariate analyses were used to characterize the literacy and cognitive skills of the overall sample and logistic regression analysis was used to test the relation of the four groups to non-adherence (< 95%). In general, performance on measures of literacy and cognitive functioning were below average, with severe deficits noted in psychomotor functioning. Additionally, after adjusting for recent cocaine use, those classified as low literate/low cognition were over nine times more likely to be non-adherent than the referent high literate/high cognition group. Low literacy and cognitive impairment placed HIV-positive drug users at high risk for non-adherence, even after adjusting for recent cocaine use. The findings suggest that targeted interventions to improve these skill deficits in this population may help to improve adherence to HIV medications.


Visualization-based mapping of language function in the brain.
Cortical language maps, obtained through intraoperative electrical stimulation studies, provide a rich source of information for research on language organization. Previous studies have shown interesting correlations between the distribution of essential language sites and such behavioral indicators as verbal IQ and have provided suggestive evidence for regarding human language cortex as an organization of multiple distributed systems. Noninvasive studies using ECoG, PET, and functional MR lend support to this model; however, there as yet are no studies that integrate these two forms of information. In this paper we describe a method for mapping the stimulation data onto a 3-D MRI-based neuroanatomic model of the individual patient. The mapping is done by comparing an intraoperative photograph of the exposed cortical surface with a computer-based MR visualization of the surface, interactively indicating corresponding stimulation sites, and recording 3-D MR machine coordinates of the indicated sites. Repeatability studies were performed to validate the accuracy of the mapping technique. Six observers-a neurosurgeon, a radiologist, and four computer scientists, independently mapped 218 stimulation sites from 12 patients. The mean distance of a mapping from the mean location of each site was 2.07 mm, with a standard deviation of 1.5 mm, or within 5.07 mm with 95% confidence. Since the surgical sites are accurate within approximately 1 cm, these results show that the visualization-based approach is accurate within the limits of the stimulation maps. When incorporated within the kind of information system envisioned by the Human Brain Project, this anatomically based method will not only provide a key link between noninvasive and invasive approaches to understanding language organization, but will also provide the basis for studying the relationship between language function and anatomical variability.


Mechanical phenotyping of primary human skeletal stem cells in heterogeneous populations by real-time deformability cytometry.
Skeletal stem cells (SSCs) are a sub-population of mesenchymal stromal cells (MSCs) present in bone marrow with multipotent differentiation potential. A current unmet challenge hampering their clinical translation remains the isolation of homogeneous populations of SSCs, in vitro, with consistent regeneration and differentiation capacities. Cell stiffness has been shown to play an important role in cell separation using microfluidic techniques such as inertial focusing or deterministic lateral displacement. Here we report that the mechanical properties of SSCs, and of a surrogate human osteosarcoma cell line (MG-63), differ significantly from other cell populations found in the bone marrow. Using real-time deformability cytometry, a recently introduced method for cell mechanical characterization, we demonstrate that both MG-63 and SSCs are stiffer than the three primary leukocyte lineages (lymphocytes, monocytes and granulocytes) and also stiffer than HL-60, a human leukemic progenitor cell line. In addition, we show that SSCs form a mechanically distinct sub-population of MSCs. These results represent an important step towards finding the bio-physical fingerprint of human SSCs that will allow their label-free separation from bone marrow with significant physiological and therapeutic implications.


Association of atrial nicotinamide adenine dinucleotide phosphate oxidase activity with the development of atrial fibrillation after cardiac surgery.
Our goal was to evaluate the role of myocardial nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity and plasma markers of oxidative stress in the pathogenesis of post-operative atrial fibrillation (AF). Atrial fibrillation is a common complication of cardiac surgery, leading to increased morbidity and prolonged hospitalization. Experimental evidence suggests that oxidative stress may be involved in the pathogenesis of AF; however, the relevance of this putative mechanism in patients undergoing cardiac surgery is unclear. We measured basal and NADPH-stimulated superoxide production in right atrial appendage samples from 170 consecutive patients undergoing conventional coronary artery bypass surgery. Plasma markers of lipid and protein oxidation (thiorbabituric acid-reactive substances, 8-isoprostane, and protein carbonyls) were also measured in blood samples drawn from a central line before surgery and after reperfusion. Patients who developed AF after surgery (42%) were older and had a significantly increased atrial NADPH oxidase activity than patients who remained in sinus rhythm (SR) (in relative light units/s/mug protein: 4.78 +/- 1.44 vs. 3.53 +/- 1.04 in SR patients, p < 0.0001). Plasma markers of lipid and protein oxidation increased significantly after reperfusion; however, neither pre-operative nor post-operative measurements differed between patients who developed AF and those who remained in SR after surgery. Multivariate analysis identified atrial NADPH oxidase activity as the strongest independent predictor of post-operative AF (odds ratio 2.41; 95% confidence interval 1.71 to 3.40, p < 0.0001). Atrial NADPH oxidase activity is independently associated with an increased risk of post-operative AF, suggesting that this oxidase system may be a key mediator of atrial oxidative stress leading to the development of AF after cardiac surgery.


Colonization of a Deglaciated Moraine: Contrasting Patterns of Carbon Uptake and Release from C3 and CAM Plants.
Current glacier retreat makes vast mountain ranges available for vegetation establishment and growth. As a result, carbon (C) is accumulated in the soil, in a negative feedback to climate change. Little is known about the effective C budget of these new ecosystems and how the presence of different vegetation communities influences CO2 fluxes. On the Matsch glacier forefield (Alps, Italy) we measured over two growing seasons the Net Ecosystem Exchange (NEE) of a typical grassland, dominated by the C3 Festuca halleri All., and a community dominated by the CAM rosettes Sempervivum montanum L. Using transparent and opaque chambers, with air temperature as the driver, we partitioned NEE to calculate Ecosystem Respiration (Reco) and Gross Ecosystem Exchange (GEE). In addition, soil and vegetation samples were collected from the same sites to estimate the Net Ecosystem Carbon Balance (NECB). The two communities showed contrasting GEE but similar Reco patterns, and as a result they were significantly different in NEE during the period measured. The grassland acted as a C sink, with a total cumulated value of -46.4±35.5 g C m-2 NEE, while the plots dominated by the CAM rosettes acted as a source, with 31.9±22.4 g C m-2. In spite of the different NEE, soil analysis did not reveal significant differences in carbon accumulation of the two plant communities (1770±130 for F. halleri and 2080±230 g C m-2 for S. montanum), suggesting that processes often neglected, like lateral flows and winter respiration, can have a similar relevance as NEE in the determination of the Net Ecosystem Carbon Balance.


RP-CARS: label-free optical readout of the myelin intrinsic healthiness.
Here we present a method based on Rotating-Polarization Coherent Anti-Stokes Raman Scattering (RP-CARS) imaging to assess the myelin health status in mouse sciatic nerves. Differently from the existing techniques, our method is based on the readout of intrinsic molecular architecture rather than on the image analysis, relying on the fact that healthy myelin is characterized by a high degree of molecular order. We exploit RP-CARS imaging to demonstrate that the degree of spatial anisotropy of the CARS signal displays a strong correlation with the g-ratio (a well-known image-based index of myelin damage) in a chemical-damage model and therefore that the former is a good indicator for the local myelin health status.