Engraftment of gene-marked hematopoietic progenitors in myeloma patients after transplant of autologous long-term marrow cultures.
We conducted a phase I hematopoietic stem cell (HSC) gene-marking trial in patients undergoing autologous blood or marrow stem cell transplant for the treatment of multiple myeloma. Between 500 and 1000 ml of bone marrow was harvested from each of 14 myeloma patients and 1 syngeneic donor. A mean of 3.3x10(9) cells per patient were plated in 20 to 50 long-term marrow culture (LTMC) flasks and maintained for 3 weeks. LTMCs were exposed on days 8 and 15 to clinical-grade neo(r)-containing retrovirus supernatant (G1Na). A mean of 8.23x10(8) day-21 LTMC cells containing 5.2x10(4) gene-marked granulocyte-macrophage progenitor cells (CFU-GM) were infused along with an unmanipulated peripheral blood stem cell graft into each patient after myeloablative therapy. Proviral DNA was detected in 71% of 68 tested blood and bone marrow samples and 150 of 2936 (5.1%) CFU-GM derived from patient bone marrow samples after transplant. The proportion of proviral DNA-positive CFU-GM declined from a mean of 9.8% at 3 months to a mean of 2.3% at 24 months postinfusion. Southern blots of 26 marrow and blood samples were negative. Semiquantitative PCR analysis indicated that gene transfer was achieved in 0.01-1% of total bone marrow and blood mononuclear cells (MNCs). Proviral DNA was also observed in EBV-transformed B lymphocytes, in CD34+ -enriched bone marrow cells, and in CFUs derived from the latter progenitors. Gene-modified cells were detected by PCR in peripheral blood and bone marrow for 24 months after infusion of LTMC cells. Sensitivity and specificity of the PCR assays were independently validated in four laboratories. Our data confirm that HSCs may be successfully transduced in stromal based culture systems. The major obstacle to therapeutic application of this approach remains the overall low level of genetically modified cells among the total hematopoietic cell pool in vivo.


Sexual selection and ecological generalism are correlated in antbirds.
Sexual selection is thought to counteract natural selection on the grounds that secondary sexual traits are inherently costly and evolve at the expense of naturally selected traits. It is therefore commonly predicted that increased sexual selection is associated with decreased physiological tolerance or ecological plasticity. Using phylogenetic comparative methods, we test this prediction by exploring relationships between traits assumed to be sexually selected (plumage dichromatism and song structure) and traits assumed to be naturally selected (altitudinal range and habitat range) in a diverse family of tropical birds. Contrary to expectations, we find that taxa with higher levels of dichromatism, and lower song pitch, occupy a wider variety of habitats and elevations. In other words, indices of sexual selection are positively related to two standard measures of ecological generalism. One interpretation of this pattern is that sexual selection combines synergistically with natural selection, thereby increasing physiological tolerance or the propensity to adapt to novel environments. An alternative possibility is that ecological generalism increases population density, which in turn promotes sexual selection in the form of greater competition for mates. Overall, our results suggest that a synergism between natural selection and sexual selection may be widespread, but the processes underlying this pattern remain to be investigated.


Impact of the European Working Time Directive (EWTD) on the operative experience of surgery residents.
In Europe and the United States, work hour restrictions are considered to be particularly burdensome for residents in surgery specialties. The aim of this study was to examine whether reduction of the work week to 48 hours resulting from the implementation of the European Working Time Directive has affected the operative experience of surgery residents. This study was conducted in a general surgery training region in the Netherlands, consisting of 1 university hospital and 6 district training hospitals. Operating records summarizing the surgical procedures performed as "primary surgeon" in the operating theater for different grades of surgeons were retrospectively analyzed for the period 2005-2012 by the use of linear regression models. Operative procedures performed by residents were considered the main outcome measure. In total, 235,357 operative procedures were performed, including 47,458 (20.2%) in the university hospital and 187,899 (79.8%) in the district training hospitals (n = 5). For residents in the university hospital, the mean number of operative procedures performed per 1.0 full-time equivalent increased from 128 operations in 2005 to 204 operations in 2012 (P = .001), whereas for residents in district training hospitals, no substantial differences were found over time. The mean (±SD) operative caseload of 64 residents who completed the 6-year training program between 2005 and 2012 was 1,391 ± 226 (range, 768-1856). A comparison of the operative caseload according to year of board-certification showed no difference. Implementation of the European Working Time Directive has not affected adversely the number of surgical procedures performed by residents within a general surgical training region in the Netherlands.


[The effect of cervical arthrosis in the etiopathogenesis of vertebrobasilar insufficiency].
We studied four groups of one hundred patients older than forty years. A group with radiological signs of cervical arthrosis was statistically compared with another one without such signs. There was no significant difference between the two groups with regard to the presence of vertebrobasilar insufficiency. Then we compared one hundred patients diagnosed of vertebrobasilar insufficiency with another group without this disease. When cervical arthrosis cases in these two groups were statistically compared, we could not find any significant differences. We think cervical arthrosis is not an important etiologic factor in vertebrobasilar insufficiency. Our results point to atherosclerotic lesions as the main cause of this entity.


Clinical Value of a Nomogram Model Based on Apparent Diffusion Coefficient Values Within 1 cm of the Tumor Cavity to Predict Postoperative Progression of Glioma.
To explore the clinical value of constructing a nomogram model based on apparent diffusion coefficient values within 1 cm of the residual tumor cavity to predict the postoperative progression of gliomas. Clinical data of patients with glioma who underwent surgery were retrospectively retrieved from the First Hospital of Qinhuangdao. The mean apparent diffusion coefficient (mADC) was measured using a picture archiving and communication system. The Kaplan-Meier survival curve was constructed with the optimal mADC threshold determined by the X-tile. A nomogram was developed based on the independent risk factors determined using the Cox proportional hazards model (Cox regression model) to predict the progression of postoperative glioma. A receiver operating characteristic curve was drawn to evaluate the prediction accuracy of the model, and decision curve analysis was performed to assess the clinical value of the nomogram. There was good agreement between the mADC values of the 2 repeated measurements before and after, with a consistency correlation coefficient of 0.83. Multivariate Cox regression analysis showed that peritumoral mADC values, degree of peritumoral enhancement, age, pathological grading, and degree of tumor resection were independent risk factors for predicting postoperative progression of glioma (all P < 0.05). The receiver operating characteristic curves of the nomogram predicting 1, 2, and 3 years postoperative progression were 0.86, 0.82, and 0.91, respectively. The calibration curve showed good consistency between the observed and predicted values in the model. The curve showed that the nomogram model has a good clinical application value. The peritumoral mADC values, degree of peritumoral enhancement, age, pathological grade, and degree of tumor resection were independent factors affecting the postoperative progression of glioma. The nomogram model established for the first time based on mADC values within 1 cm of the tumor can predict the postoperative condition of patients with glioma intuitively and comprehensively. It can provide a relatively accurate prediction tool for neurosurgeons to individualize the evaluation of survival and prognosis, and formulate treatment plans for patients.


In vivo magnetic resonance imaging of injected endothelial progenitor cells after myocardial infarction in rats.
The purpose of this study was to detect and follow transplanted superparamagnetic iron oxide (SPIO)-labeled endothelial progenitor cells (EPCs) by magnetic resonance imaging (MRI). Infarcted rats were randomized to injections of SPIO-labeled EPCs, unlabeled EPCs, or saline. From 1 day to 8 weeks, in vivo serial MRI was performed for cell tracking. Labeled cells were visualized as hypointense areas by MRI. The presence of labeled EPCs at 10 days and disappearance of these cells by 8 weeks was confirmed by iron and 4',6-diamidino-2-phenylindole. Co-staining for iron and ED-1 showed that the iron-positive cells were macrophages. EPC implantation significantly elevated vascular endothelial growth factor expression, accompanied by increased capillary and arteriole density in the ischemic myocardium. At 8 weeks, the transplanted EPCs were not present and the enhanced MRI signals arose from macrophages. However, both EPCs enhanced cardiac function. The major mechanism of cardiac improvement appears to be paracrine pathways of the engrafted EPCs.


PepNovo: de novo peptide sequencing via probabilistic network modeling.
We present a novel scoring method for de novo interpretation of peptides from tandem mass spectrometry data. Our scoring method uses a probabilistic network whose structure reflects the chemical and physical rules that govern the peptide fragmentation. We use a likelihood ratio hypothesis test to determine whether the peaks observed in the mass spectrum are more likely to have been produced under our fragmentation model than under a model that treats peaks as random events. We tested our de novo algorithm PepNovo on ion trap data and achieved results that are superior to popular de novo peptide sequencing algorithms. PepNovo can be accessed via the URL http://www-cse.ucsd.edu/groups/bioinformatics/software.html.


Principal manifolds and graphs in practice: from molecular biology to dynamical systems.
We present several applications of non-linear data modeling, using principal manifolds and principal graphs constructed using the metaphor of elasticity (elastic principal graph approach). These approaches are generalizations of the Kohonen's self-organizing maps, a class of artificial neural networks. On several examples we show advantages of using non-linear objects for data approximation in comparison to the linear ones. We propose four numerical criteria for comparing linear and non-linear mappings of datasets into the spaces of lower dimension. The examples are taken from comparative political science, from analysis of high-throughput data in molecular biology, from analysis of dynamical systems.


Current Challenges in the Recognition and Management of Delirium Superimposed on Dementia.
Delirium occurring in a patient with preexisting dementia is referred to as delirium superimposed on dementia (DSD). DSD commonly occurs in older hospitalized patients and is associated with worse outcomes, including higher rates of mortality and institutionalization, compared to inpatients with delirium or dementia alone. This narrative review summarizes the screening, diagnosis, management, and pathophysiology of DSD and concludes by highlighting opportunities for future research. Studies were identified via Medline and PsycINFO keyword search, and handsearching reference lists. Conceptually, DSD could be considered an "acute exacerbation" of dementia precipitated by a noxious insult akin to an acute exacerbation of heart failure or acute on chronic renal failure. However, unlike other organ systems, there are no established biomarkers for delirium, so DSD is diagnosed and monitored clinically. Because cognitive dysfunction is common to both delirium and dementia, the diagnosis of DSD can be challenging. Inattention, altered levels of arousal, and motor dysfunction may help distinguish DSD from dementia alone. An informant history suggestive of an acute change in cognition or alertness should be investigated and managed as delirium until proven otherwise. The key management principles include prevention, identifying and treating the underlying precipitant(s), implementing multicomponent interventions to create an ideal environment for brain recovery, preventing complications, managing distress, and monitoring for resolution. Informing and involving family members or caregivers throughout the patient journey are essential because there is significant prognostic uncertainty, including the risk of persistent cognitive and functional decline following DSD and relapse. Furthermore, informal carers can provide significant assistance in management. Emerging evidence demonstrates that increased exposure to delirium is associated with neuronal injury and worse cognitive outcomes although the mechanisms through which this occurs remain unclear. Given the clinical overlap between delirium and dementia, studying shared pathophysiological pathways may uncover diagnostic tests and is an essential step in therapeutic innovation.


Stomatocytic haemolysis and macrothrombocytopenia (Mediterranean stomatocytosis/macrothrombocytopenia) is the haematological presentation of phytosterolaemia.
Phytosterolaemia (sitosterolaemia) is a recessively inherited metabolic condition in which the absorption of both cholesterol and plant-derived cholesterol-like molecules at the gut is unselective and unrestricted. In haematology, Mediterranean stomatocytosis or Mediterranean macrothrombocytopenia is a poorly understood haematological condition that combines stomatocytic haemolysis with the presence of very large platelets. Five pedigrees showing this haematology were identified. Gas chromatography mass spectrometry (GC-MS) showed that all of the patients with this highly specific haematology had grossly elevated levels of phytosterols in the blood, diagnostic of phytosterolaemia. All showed mutations in the ABCG5 and ABCG8 previously linked to phytosterolaemia. Three pedigrees showed five new mutations, while two pedigrees showed the common W361X mutation in ABCG8. We draw the following four conclusions: (i) that Mediterranean stomatocytosis/macrothrombocytopenia is caused by an excess of phytosterols in the blood; (ii) that phytosterolaemia, which does not respond to standard statin treatment, can be diagnosed via the distinctive haematology described here, even when the cholesterol is normal; (iii) that phytosterolaemia should be considered in the differential diagnosis of all patients with large platelets; and (iv) that the platelet size should be noted in patients with hypercholesterolaemia.


Machine Learning-Based Mortality Prediction of Patients at Risk During Hospital Admission.
The ability to predict in-hospital mortality from data available at hospital admission would identify patients at risk and thereby assist hospital-wide patient safety initiatives. Our aim was to use modern machine learning tools to predict in-hospital mortality from standardized data sets available at hospital admission. This was a retrospective, observational study in 3 adult tertiary care hospitals in Western Australia between January 2008 and June 2017. Primary outcome measures were the area under the curve for the receiver operating characteristics curve, the F1 score, and the average precision of the 4 machine learning algorithms used: logistic regression, neural networks, random forests, and gradient boosting trees. Using our 4 predictive models, in-hospital mortality could be predicted satisfactorily (areas under the curve for neural networks, logistic regression, random forests, and gradient boosting trees: 0.932, 0.936, 0.935, and 0.935, respectively), with moderate F1 scores: 0.378, 0.367, 0.380, and 0.380, respectively. Average precision values were 0.312, 0.321, 0.334, and 0.323, respectively. It remains unknown whether additional features might improve our models; however, this would result in additional efforts for data acquisition in daily clinical practice. This study demonstrates that using only a limited, standardized data set in-hospital mortality can be predicted satisfactorily at the time point of hospital admission. More parameters describing patient's health are likely needed to improve our model.


MTHFD2 is a metabolic checkpoint controlling effector and regulatory T cell fate and function.
Antigenic stimulation promotes T cell metabolic reprogramming to meet increased biosynthetic, bioenergetic, and signaling demands. We show that the one-carbon (1C) metabolism enzyme methylenetetrahydrofolate dehydrogenase 2 (MTHFD2) regulates de novo purine synthesis and signaling in activated T cells to promote proliferation and inflammatory cytokine production. In pathogenic T helper-17 (Th17) cells, MTHFD2 prevented aberrant upregulation of the transcription factor FoxP3 along with inappropriate gain of suppressive capacity. MTHFD2 deficiency also promoted regulatory T (Treg) cell differentiation. Mechanistically, MTHFD2 inhibition led to depletion of purine pools, accumulation of purine biosynthetic intermediates, and decreased nutrient sensor mTORC1 signaling. MTHFD2 was also critical to regulate DNA and histone methylation in Th17 cells. Importantly, MTHFD2 deficiency reduced disease severity in multiple in vivo inflammatory disease models. MTHFD2 is thus a metabolic checkpoint to integrate purine metabolism with pathogenic effector cell signaling and is a potential therapeutic target within 1C metabolism pathways.


Enantioselective Amination of 4-Substituted Pyrazolones Catalyzed by Oxindole-Containing Thioureas and by a Recyclable Linear-Polymer-Supported Analogue in a Continuous Flow Process.
A highly efficient organocatalytic amination of 4-substituted pyrazolones with azodicarboxylates mediated by a novel quinine-derived thiourea with a 3,3-diaryl-oxindole scaffold is reported. This synthetic method furnished 4-amino-5-pyrazolones in high yields and with excellent enantioselectivities (up to 97:3 er) at room temperature in short reaction times. Moreover, a linear-polymer-supported bifunctional thiourea, synthesized by reacting a bifunctional aromatic monomer (biphenyl) with isatin in superacidic media and further derivatization, was proven to be also an efficient heterogeneous organocatalyst for this α-amination reaction. The practical value of this process was demonstrated by the use of the immobilized catalyst in recycling experiments, maintaining the activity without additional reactivation, and in flow processes, allowing the synthesis of 4-amino-pyrazolone derivatives in a gram scale with high yield and enantioselectivity.


Effect of oxygen and shear stress on molecular weight of hyaluronic acid.
Dissolved oxygen (DO) and shear stress have pronounced effects on hyaluronic acid (HA) production, yet various views persist about their effect on the molecular weight of HA. Accordingly, this study investigated the effects of DO and shear stress during HA fermentation. The results showed that both cell growth and HA synthesis were suppressed under anaerobic conditions, and the HA molecular mass was only (1.22+/-0.02) x 106 Da. Under aerobic conditions, although the DO level produced no change in the biomass or HA yield, a high DO level favored the HA molecular mass, which reached a maximum value of (2.19+/- 0.05) x 106 Da at 50% DO. Furthermore, a high shear stress delayed the rate of HA synthesis and decreased the HA molecular weight, yet had no clear effect on the HA yield. Therefore, a high DO concentration and mild shear environment would appear to be essential to enhance the HA molecular weight.


Beyond cycle lanes and large-scale infrastructure: a scoping review of initiatives that groups and organisations can implement to promote cycling for the Cycle Nation Project.
Cycling has well-established positive relationships with health. Evidence suggests that large-scale infrastructure and built-environment initiatives to promote cycling are likely to be necessary but not sufficient to maximise cycling participation. Smaller-scale initiatives that can be implemented by organisations (eg, employers) and groups (eg, community groups) are therefore also important, but the full range of feasible activities to promote cycling is not known. We aimed to scope the literature and map organisational, social and individual level activities to increase cycling. Design: Scoping review following an established five-stage process.Eligibility criteria: Studies or publicly available reports describing cycling promotion initiatives deemed feasible for organisations or groups to implement.Sources of evidence and selection: (i) online databases (Ovid (Medline), Ovid (Embase), SportDISCUS (Ebscohost), ProQuest, Web of Science), (ii) existing systematic reviews, (iii) expert stakeholder consultation. We extracted data from 129 studies and reports, from 20 different countries, identifying 145 cycling promotion initiatives. From these initiatives we identified 484 actions within 93 action types within 33 action categories under the nine intervention functions described by Michie et al. Environmental restructuring (micro-level), enablement, education and persuasion were the functions with the most action types, while coercion, modelling and restriction had the fewest action types. This is the first comprehensive map to summarise the broad range of action types feasible for implementation within organisation/group-based cycling promotion initiatives. The map will be a critical tool for communities, employers, practitioners and researchers in designing interventions to increase cycling.


NuA4 and H2A.Z control environmental responses and autotrophic growth in Arabidopsis.
Nucleosomal acetyltransferase of H4 (NuA4) is an essential transcriptional coactivator in eukaryotes, but remains poorly characterized in plants. Here, we describe Arabidopsis homologs of the NuA4 scaffold proteins Enhancer of Polycomb-Like 1 (AtEPL1) and Esa1-Associated Factor 1 (AtEAF1). Loss of AtEAF1 results in inhibition of growth and chloroplast development. These effects are stronger in the Atepl1 mutant and are further enhanced by loss of Golden2-Like (GLK) transcription factors, suggesting that NuA4 activates nuclear plastid genes alongside GLK. We demonstrate that AtEPL1 is necessary for nucleosomal acetylation of histones H4 and H2A.Z by NuA4 in vitro. These chromatin marks are diminished genome-wide in Atepl1, while another active chromatin mark, H3K9 acetylation (H3K9ac), is locally enhanced. Expression of many chloroplast-related genes depends on NuA4, as they are downregulated with loss of H4ac and H2A.Zac. Finally, we demonstrate that NuA4 promotes H2A.Z deposition and by doing so prevents spurious activation of stress response genes.


Particle-Polymer Union with Changeable Wettability for Constructing Bijels Using a Simple Mixing Method.
Bicontinuous emulsion gels (bijels) are nonequilibrium dispersed systems with particle-stabilized continuous fluid domains, and the internal connectivity of channels brings the possibility of efficient mass transport, endowing bijels great potential in diverse applications. Different from the common method to produce bijels, the spinodal decomposition, which needs precise temperature control and is restricted by the selection of liquid pairs, in this work, a direct mixing method was performed to construct bijels, simplifying the fabrication process. The hydrophilic rod-shaped cellulose nanocrystalline (CNC) particles were in situ combined with the hydrophobic polymer, aminopropyl-terminated polydimethylsiloxane (PDMS-NH2), to acquire a controllable interfacial wettability of CNC. The CNC@mPDMS-NH2 complexes were adsorbed at the water-toluene interface and achieved a change of Pickering emulsion types, oil-in-water, bijel, and water-in-oil, through tuning the interfacial performance of CNC@mPDMS-NH2 complexes. A three-dimensional scanning image and curvature calculation were applied to verify the obtained bijel, further demonstrating the successful preparation of the bicontinuous structure. This work enriched the members of particles for stabilizing bijels and was considered to be scalable in manufacturing for applications on a large scale.


[Surgery of the juvenile adrenal glands. Surgical protocol, intra- and perioperative monitoring].
Surgical and non-surgical conditions of the adrenal cortex and medulla are presented. Besides general principles of operative treatment (e.g. surgical approach, safe operative technique, bilateral exploration) specific aspects associated with this procedure in childhood are discussed. The perioperative management and pharmacotherapeutical preparation (e.g. hormonal substitution) are emphasized.


[Concept and classification of obesity].
Obesity is recognized as a disease when it is associated with current or future health problems. The association of obesity with hypertension, glucose intolerance, and lipid metabolism disorder is diagnosed as metabolic syndrome since it tends to cause arteriosclerotic diseases. Obesity is important since it is pandemic throughout the earth. In this review we explain the concept and classification of obesity. We discuss the disease basis of obesity and metabolic syndrome, especially from the standpoint of adipotoxicity.


Characterization of the kaurene oxidase CYP701A3, a multifunctional cytochrome P450 from gibberellin biosynthesis.
KO (kaurene oxidase) is a multifunctional cytochrome P450 catalysing three sequential oxidations in gibberellin phytohormone biosynthesis. These serve to transform the C4α methyl of the ent-kaurene olefin intermediate into the carboxylic acid moiety of ent-kauren-19-oic acid. To investigate the unknown catalytic mechanism and properties of KO, we have engineered the corresponding CYP701A3 from Arabidopsis thaliana (AtKO) for functional recombinant expression in Escherichia coli, involving use of a fully codon-optimized construct, along with additional N-terminal deletion and modification. This recombinant AtKO (rAtKO) was used to carry out 18O2 labelling studies with ent-kaurene, and the intermediates ent-kaurenol and ent-kaurenal, to investigate the multifunctional reaction sequence; revealing catalysis of three hydroxylation reactions, which further requires dehydration at some stage. Accordingly, following initial hydroxylation, ent-kaurenol must then be further hydroxylated to a gem-diol intermediate, and our data indicate that the subsequent reactions proceed via dehydration of the gem-diol to ent-kaurenal, followed by an additional hydroxylation to directly form ent-kaurenoic acid. Kinetic analysis indicates that these intermediates are all retained in the active site during the course of the reaction series, with the first hydroxylation being rate-limiting. In addition, investigation of alternative substrates demonstrated that ent-beyerene, which differs in ring structure distal to the C4α methyl, is only hydroxylated by rAtKO, indicating the importance of the exact tetracyclic ring structure of kaurane for multifunctional KO activity. Thus the results of the present study clarify the reaction sequence and enzymatic mechanism of KO, as well as substrate features critical for the catalysed multiple reaction sequence.


Emulating short-term synaptic dynamics with memristive devices.
Neuromorphic architectures offer great promise for achieving computation capacities beyond conventional Von Neumann machines. The essential elements for achieving this vision are highly scalable synaptic mimics that do not undermine biological fidelity. Here we demonstrate that single solid-state TiO2 memristors can exhibit non-associative plasticity phenomena observed in biological synapses, supported by their metastable memory state transition properties. We show that, contrary to conventional uses of solid-state memory, the existence of rate-limiting volatility is a key feature for capturing short-term synaptic dynamics. We also show how the temporal dynamics of our prototypes can be exploited to implement spatio-temporal computation, demonstrating the memristors full potential for building biophysically realistic neural processing systems.


Neural discriminant analysis.
In this article the role of the bootstrap is highlighted for nonlinear discriminant analysis using a feedforward neural network model. Statistical techniques are formulated in terms of the principle of the likelihood of a neural-network model when the data consist of ungrouped binary responses and a set of predictor variables. We illustrate that the information criterion based on the bootstrap method is shown to be favorable when selecting the optimum number of hidden units for a neural-network model. In order to summarize the measure of goodness-of-fit, the deviance on fitting a neural-network model to binary response data can be bootstrapped. We also provide the bootstrap estimates of the biases of excess error in a prediction rule constructed by fitting to the training sample in the neural network model. We additionally propose bootstrap methods for the analysis of residuals in order to identify outliers and examine distributional assumptions in neural-network model fitting. These methods are illustrated through the analyzes of medical diagnostic data.


Dietary (n-3) polyunsaturated fatty acids do not affect the in vivo development and function of Listeria-specific CD4+ and CD8+ effector and memory/effector T cells in mice.
We previously reported that in a mouse model, a diet high in (n-3) PUFA diminishes host survival following an infection from Listeria monocytogenes, a gram-positive bacterial pathogen. In this study we investigated the impact of (n-3) PUFA on the adaptive immune response to L. monocytogenes. BALB/c mice were fed experimental diets either devoid of or rich in (n-3) PUFA from fish oil for 4 wk and then infected with 10(6) actA-deficient L. monocytogenes. At 7 and 35 d postchallenge, effector and memory/effector T cells in the spleen were enumerated by flow cytometry. Surprisingly, the number of Listeria-specific CD4(+) and CD8(+) effector and memory/effector T cells in the spleen was not affected by (n-3) PUFA. Also, the effector cells derived from mice fed either diet were equally capable of conferring protective immunity upon adoptive transfer to naive recipients. Despite our previous data, which demonstrated that (n-3) PUFA profoundly impaired host resistance to L. monocytogenes, pathogen-specific T cell responses were not substantially affected by dietary (n-3) PUFA.


Lipidomics reveals insights on the biological effects of copper oxide nanoparticles in a human colon carcinoma cell line.
Engineered nanomaterials have unique properties compared to their bulk counterparts. Copper oxide nanoparticles (CuO NPs) are one example of nanomaterials used in a wide range of consumer products due to their conductivity and biocidal properties. While CuO NPs can induce toxicity in various organisms, their interactions with different organisms and how they affect cellular homeostasis is yet to be fully understood. In this work, the toxicity of CuO NPs was evaluated in different human cell lines (colorectal carcinoma, cervical cancer, embryonic kidney, and lung fibroblast), showing a dose-dependent toxicity. An untargeted lipidomics approach using liquid chromatography-quadrupole time of flight mass spectrometry was employed in a human colon carcinoma cell line to investigate the impact of CuO NP exposure at the cellular level. A 24 h CuO NP exposure at 2.5 and 5 μg mL-1 resulted in upregulation of different metabolites: triacylglycerols, phosphatidylcholines, and ceramides accumulated. The most profound increase in a dose-dependent manner was observed in ceramides, specifically in C18:0, C18:1, and C22:0 species, with up to ∼10 fold accumulations. Further experiments suggested that activation of autophagy and oxidative stress could be responsible for the toxicity observed in these cell lines. Increases in the level of glutathione oxide (∼7 fold) also supported the activation of oxidative stress upon CuO NP treatment. Based on the changes in different metabolites induced by CuO NP exposure and previous studies from our laboratory, we propose that autophagy and oxidative stress could play a role in CuO NP-induced toxicity.


The effect of tylosin on antimicrobial resistance in beef cattle enteric bacteria: A systematic review and meta-analysis.
Tylosin is a commonly used in-feed antimicrobial and is approved in several countries to reduce the incidence of liver abscesses in beef cattle. Macrolides are critically important antimicrobials in human health and used to treat some foodborne bacterial diseases, such as Campylobacter jejuni and Salmonella. Feeding tylosin could select for resistant enteric bacteria in cattle, which could contaminate beef products at slaughter and potentially cause foodborne illness. We conducted a systematic review and meta-analysis to evaluate the impact of feeding tylosin to cattle on phenotypic and genotypic resistance in several potential zoonotic enteric bacteria: Enterococcus species, Escherichia coli, Salmonella enterica subspecies enterica, and Campylobacter species. This review was registered with PROSPERO (#CRD42018085949). Eleven databases were searched for primary research studies that fed tylosin at approved doses to feedlot cattle and tested bacteria of interest for phenotypic or genotypic resistance. We screened 1,626 citations and identified 13 studies that met the inclusion criteria. Enterococcus species were tested in seven studies, Escherichia coli was isolated in five studies, three studies reported on Salmonella, and two studies reported on Campylobacter species. Most studies relied on phenotypic antimicrobial susceptibility testing and seven also reported resistance gene testing. A random-effects meta-analyses of erythromycin-resistant enterococci from four studies had significant residual heterogeneity. Only two studies were available for a meta-analysis of tylosin-resistant enterococci. A semi-quantitative analysis demonstrated an increase in macrolide-resistant enterococci after long durations of tylosin administration (>100 days). Semi-quantitative analyses of other bacteria-antimicrobial combinations revealed mixed results, but many comparisons found no effect of tylosin administration. However, about half of these no-effect comparisons did not record the cumulative days of tylosin administration or the time since the last dose. When fed at approved dosages for typical durations, tylosin increases the proportion of macrolide-resistant enterococci in the cattle gastrointestinal tract, which could pose a zoonotic risk to human beef consumers. Feeding tylosin for short durations may mitigate the impact on macrolide-resistant enterococci and further studies are encouraged to determine the effect of minimizing or eliminating tylosin use in beef cattle. There may also be an impact on other bacteria and other antimicrobial resistances but additional details or data are needed to strengthen these comparisons. We encourage authors of antimicrobial-resistance studies to follow reporting guidelines and publish details of all comparisons to strengthen future meta-analyses.


Variation in brain organization of coral reef fish larvae according to life history traits.
In coral reefs, one of the great mysteries of teleost fish ecology is how larvae locate the relatively rare patches of habitat to which they recruit. The recruitment of fish larvae to a reef, after a pelagic phase lasting between 10 and 120 days, depends strongly on larval ability to swim and detect predators, prey and suitable habitat via sensory cues. However, no information is available about the relationship between brain organization in fish larvae and their sensory and swimming abilities at recruitment. For the first time, we explore the structural diversity of brain organization (comparative sizes of brain subdivisions: telencephalon, mesencephalon, cerebellum, vagal lobe and inferior lobe) among larvae of 25 coral reef fish species. We then investigate links between variation in brain organization and life history traits (swimming ability, pelagic larval duration, social behavior, diel activity and cue use relying on sensory perception). After accounting for phylogeny with independent contrasts, we found that brain organization covaried with some life history traits: (1) fish larvae with good swimming ability (>20 cm/s), a long pelagic duration (>30 days), diurnal activity and strong use of cues relying on sensory perception for detection of recruitment habitat had a larger cerebellum than other species. (2) Fish larvae with a short pelagic duration (<30 days) and nocturnal activity had a larger mesencephalon and telencephalon. Lastly, (3) fish larvae exhibiting solitary behavior during their oceanic phase had larger inferior and vagal lobes. Overall, we hypothesize that a well-developed cerebellum may allow fish larvae to improve their chances of successful recruitment after a long pelagic phase in the ocean. Our study is the first one to bring together quantitative information on brain organization and the relative development of major brain subdivisions across coral reef fish larvae, and more specifically to address the way in which this variation correlates with the recruitment process.


Psychometrican analysis and dimensional structure of the Brazilian version of melasma quality of life scale (MELASQoL-BP).
Although asymptomatic, melasma inflicts significant impact on quality of life. MELASQoL is the main instrument used to assess quality of life associated with melasma, it has been validated in several languages, but its latent dimensional structure and psychometric properties haven´t been fully explored. To evaluate psychometric characteristics, information and dimensional structure of the Brazilian version of MELASQoL. Survey with patients with facial melasma through socio-demographic questionnaire, DLQI-BRA, MASI and MELASQoL-BP, exploratory and confirmatory factor analysis, internal consistency of MELASQoL and latent dimensions (Cronbach's alpha). The informativeness of the model and items were investigated by the Rasch model (ordinal data). We evaluated 154 patients, 134 (87%) were female, mean age (± SD) of 39 (± 8) years, the onset of melasma at 27 (± 8) years, median (p25-p75) of MASI scores , DLQI and MELASQoL 8 (5-15) 2 (1-6) and 30 (17-44). The correlation (rho) of MELASQoL with DLQI and MASI were: 0.70 and 0.36. Exploratory factor analysis identified two latent dimensions: Q1-Q3 and Q4-Q10, which had significantly more adjusted factor structure than the one-dimensional model: Χ2 / gl = 2.03, CFI = 0.95, AGFI = 0.94, RMSEA = 0.08. Cronbach's coefficient for the one-dimensional model and the factors were: 0.95, 0.92 and 0.93. Rasch analysis demonstrated that the use of seven alternatives per item resulted in no increase in the model informativeness. MELASQoL-BP showed good psychometric performance and a latent structure of two dimensions. We also identified an oversizing of item alternatives to characterize the aggregate information to each dimension.


Comprehensive proteomic analysis of bovine spermatozoa of varying fertility rates and identification of biomarkers associated with fertility.
Male infertility is a major problem for mammalian reproduction. However, molecular details including the underlying mechanisms of male fertility are still not known. A thorough understanding of these mechanisms is essential for obtaining consistently high reproductive efficiency and to ensure lower cost and time-loss by breeder. Using high and low fertility bull spermatozoa, here we employed differential detergent fractionation multidimensional protein identification technology (DDF-Mud PIT) and identified 125 putative biomarkers of fertility. We next used quantitative Systems Biology modeling and canonical protein interaction pathways and networks to show that high fertility spermatozoa differ from low fertility spermatozoa in four main ways. Compared to sperm from low fertility bulls, sperm from high fertility bulls have higher expression of proteins involved in: energy metabolism, cell communication, spermatogenesis, and cell motility. Our data also suggests a hypothesis that low fertility sperm DNA integrity may be compromised because cell cycle: G2/M DNA damage checkpoint regulation was most significant signaling pathway identified in low fertility spermatozoa. This is the first comprehensive description of the bovine spermatozoa proteome. Comparative proteomic analysis of high fertility and low fertility bulls, in the context of protein interaction networks identified putative molecular markers associated with high fertility phenotype.


Translation Imaging in Parkinson's Disease: Focus on Neuroinflammation.
Parkinson's disease (PD) is characterized by the loss of dopaminergic neurons in the substantia nigra pars compacta (SNpc) and the appearance of α-synuclein insoluble aggregates known as Lewy bodies. Neurodegeneration is accompanied by neuroinflammation mediated by cytokines and chemokines produced by the activated microglia. Several studies demonstrated that such an inflammatory process is an early event, and contributes to oxidative stress and mitochondrial dysfunctions. α-synuclein fibrillization and aggregation activate microglia and contribute to disease onset and progression. Mutations in different genes exacerbate the inflammatory phenotype in the monogenic compared to sporadic forms of PD. Positron Emission Tomography (PET) and Single Photon Emission Computed Tomography (SPECT) with selected radiopharmaceuticals allow in vivo imaging of molecular modifications in the brain of living subjects. Several publications showed a reduction of dopaminergic terminals and dopamine (DA) content in the basal ganglia, starting from the early stages of the disease. Moreover, non-dopaminergic neuronal pathways are also affected, as shown by in vivo studies with serotonergic and glutamatergic radiotracers. The role played by the immune system during illness progression could be investigated with PET ligands that target the microglia/macrophage Translocator protein (TSPO) receptor. These agents have been used in PD patients and rodent models, although often without attempting correlations with other molecular or functional parameters. For example, neurodegeneration and brain plasticity can be monitored using the metabolic marker 2-Deoxy-2-[18F]fluoroglucose ([18F]-FDG), while oxidative stress can be probed using the copper-labeled diacetyl-bis(N-methyl-thiosemicarbazone) ([Cu]-ATSM) radioligand, whose striatal-specific binding ratio in PD patients seems to correlate with a disease rating scale and motor scores. Also, structural and functional modifications during disease progression may be evaluated by Magnetic Resonance Imaging (MRI), using different parameters as iron content or cerebral volume. In this review article, we propose an overview of in vivo clinical and non-clinical imaging research on neuroinflammation as an emerging marker of early PD. We also discuss how multimodal-imaging approaches could provide more insights into the role of the inflammatory process and related events in PD development.


Nonsteroidal anti-inflammatory drug-activated gene (NAG-1/GDF15) expression is increased by the histone deacetylase inhibitor trichostatin A.
Nonsteroidal anti-inflammatory drug-activated gene (NAG-1) is a putative tumor suppressor whose expression can be increased by drug treatment. Glioblastoma is the most common central nervous system tumor, is associated with high morbidity and mortality, and responds poorly to surgical, chemical, and radiation therapy. The histone deacetylase inhibitors are under current consideration as therapeutic agents in treating glioblastoma. We investigated whether trichostatin A (TSA) would alter the expression of NAG-1 in glioblastoma cells. The DNA demethylating agent 5-aza-dC did not increase NAG-1 expression, but TSA up-regulated NAG-1 expression and acted synergistically with 5-aza-dC to induce NAG-1 expression. TSA indirectly increases NAG-1 promoter activity and increases NAG-1 mRNA and protein expression in the T98G human glioblastoma cell line. TSA also increases the expression of transcription factors Sp-1 and Egr-1. Small interfering RNA experiments link NAG-1 expression to apoptosis induced by TSA. Reporter gene assays, specific inhibition by small interfering RNA transfections, and chromatin immunoprecipitation assays indicate that Egr-1 and Sp-1 mediate TSA-induced NAG-1 expression. TSA also increases the stability of NAG-1 mRNA. TSA-induced NAG-1 expression involves multiple mechanisms at the transcriptional and post-transcriptional levels.


Heritable environmental variance causes nonlinear relationships between traits: application to birth weight and stillbirth of pigs.
There is recent evidence from laboratory experiments and analysis of livestock populations that not only the phenotype itself, but also its environmental variance, is under genetic control. Little is known about the relationships between the environmental variance of one trait and mean levels of other traits, however. A genetic covariance between these is expected to lead to nonlinearity between them, for example between birth weight and survival of piglets, where animals of extreme weights have lower survival. The objectives were to derive this nonlinear relationship analytically using multiple regression and apply it to data on piglet birth weight and survival. This study provides a framework to study such nonlinear relationships caused by genetic covariance of environmental variance of one trait and the mean of the other. It is shown that positions of phenotypic and genetic optima may differ and that genetic relationships are likely to be more curvilinear than phenotypic relationships, dependent mainly on the environmental correlation between these traits. Genetic correlations may change if the population means change relative to the optimal phenotypes. Data of piglet birth weight and survival show that the presence of nonlinearity can be partly explained by the genetic covariance between environmental variance of birth weight and survival. The framework developed can be used to assess effects of artificial and natural selection on means and variances of traits and the statistical method presented can be used to estimate trade-offs between environmental variance of one trait and mean levels of others.


Picture recognition in animals and humans.
The question of object-picture recognition has received relatively little attention in both human and comparative psychology; a paradoxical situation given the important use of image technology (e. g. slides, digitised pictures) made by neuroscientists in their experimental investigation of visual cognition. The present review examines the relevant literature pertaining to the question of the correspondence between and/or equivalence of real objects and their pictorial representations in animals and humans. Two classes of reactions towards pictures will be considered in turn: acquired responses in picture recognition experiments and spontaneous responses to pictures of biologically relevant objects (e.g. prey or conspecifics). Our survey will lead to the conclusion that humans show evidence of picture recognition from an early age; this recognition is, however, facilitated by prior exposure to pictures. This same exposure or training effect appears also to be necessary in nonhuman primates as well as in other mammals and in birds. Other factors are also identified as playing a role in the acquired responses to pictures: familiarity with and nature of the stimulus objects, presence of motion in the image, etc. Spontaneous and adapted reactions to pictures are a wide phenomenon present in different phyla including invertebrates but in most instances, this phenomenon is more likely to express confusion between objects and pictures than discrimination and active correspondence between the two. Finally, given the nature of a picture (e.g. bi-dimensionality, reduction of cues related to depth), it is suggested that object-picture recognition be envisioned in various levels, with true equivalence being a limited case, rarely observed in the behaviour of animals and even humans.


Ocular hemodynamics and glaucoma prognosis: a color Doppler imaging study.
To evaluate the effect of optic nerve circulation, using color Doppler imaging (CDI), on the progression of visual field damage in primary open-angle glaucoma. The relationship between the results of retrobulbar CDI, performed shortly after the diagnosis of primary open-angle glaucoma, and the progression of visual field loss for 7 years was evaluated in 44 glaucoma patients. Color Doppler imaging variables in patients with a stable and deteriorating clinical course were compared, and the pattern of increasing risk for different CDI values was analyzed using an additive logistic model. Based on this nonparametric analysis, we arrived at a discriminant CDI value identifying glaucoma patients with a poor prognosis. On the basis of the discriminant value, patients were divided into 2 groups, and the odds ratio of visual field loss for each group was then estimated. Patients with a stable visual field had a higher diastolic velocity and a lower resistivity index in the ophthalmic artery (P<.001 for both) compared with those with a deteriorating visual field during the study. The odds of visual field deterioration in patients with an ophthalmic artery resistivity index of 0.78 or higher was about 6 times that of patients with an ophthalmic artery resistivity index lower than 0.78. Color Doppler imaging variables of the ophthalmic artery correlate with the risk of visual field deterioration in patients with primary open-angle glaucoma.


A dose-volume histogram based decision-support system for dosimetric comparison of radiotherapy treatment plans.
The choice of any radiotherapy treatment plan is usually made after the evaluation of a few preliminary isodose distributions obtained from different beam configurations. Despite considerable advances in planning techniques, such final decision remains a challenging task that would greatly benefit from efficient and reliable assessment tools. For any dosimetric plan considered, data on dose-volume histograms supplied by treatment planning systems are used to provide estimates on planning target coverage as well as on sparing of organs at risk and the remaining healthy tissue. These partial metrics are then combined into a dose distribution index (DDI), which provides a unified, easy-to-read score for each competing radiotherapy plan. To assess the performance of the proposed scoring system, DDI figures for fifty brain cancer patients were retrospectively evaluated. Patients were divided in three groups depending on tumor location and malignancy. For each patient, three tentative plans were designed and recorded during planning, one of which was eventually selected for treatment. We thus were able to compare the plans with better DDI scores and those actually delivered. When planning target coverage and organs at risk sparing are considered as equally important, the tentative plan with the highest DDI score is shown to coincide with that actually delivered in 32 of the 50 patients considered. In 15 (respectively 3) of the remaining 18 cases, the plan with highest DDI value still coincides with that actually selected, provided that organs at risk sparing is given higher priority (respectively, lower priority) than target coverage. DDI provides a straightforward and non-subjective tool for dosimetric comparison of tentative radiotherapy plans. In particular, DDI readily quantifies differences among competing plans with similar-looking dose-volume histograms and can be easily implemented for any tumor type and localization, irrespective of the planning system and irradiation technique considered. Moreover, DDI permits to estimate the dosimetry impact of different priorities being assigned to sparing of organs at risk or to better target coverage.


Understanding the structural and functional differences between mouse thyrotropin-releasing hormone receptors 1 and 2.
Multiple computational methods have been employed in a comparative study of thyrotropin-releasing hormone receptors 1 and 2 (TRH-R1 and TRH-R2) to explore the structural bases for the different functional properties of these G protein-coupled receptors. Three-dimensional models of both murine TRH receptors have been built and optimized by means of homology modeling based on the crystal structure of bovine rhodopsin, molecular dynamics simulations, and energy minimizations in a membrane-aqueous environment. The comparison between the two models showed a correlation between the higher flexibility and higher basal activity of TRH-R2 versus the lesser flexibility and lower basal activity of TRH-R1 and supported the involvement of the highly conserved W6.48 in the signaling process. A correlation between the level of basal activity and conformational changes of TM5 was detected also. Comparison between models of the wild type receptors and their W6.48A mutants, which have reversed basal activities compared with their respective wild types, further supported these correlations. A flexible molecular docking procedure revealed that TRH establishes a direct interaction with W6.48 in TRH-R2 but not in TRH-R1. We designed and performed new mutagenesis experiments that strongly supported these observations.


Alcohol dehydrogenase in maize: genetic control of enzyme activity.
A gene (Adhr(r)) which controls the activity of alcohol dehydrogenase in the scutellum of maize has been found. This gene is not allelic to the Adh(1) locus, which specifies the charge of the enzyme molecule and hence its migration rate. The two genes are linked and located about 17 crossover units apart. The Adh(r)(N) allele specifies equal activities of both the Adh(1)(S) and Adh(1)(F) products. The Adh(r)(L) allele gives lower activities of the Adh(1)(S) products only and operates in both the cis and the trans configurations.


Fragility fractures and the osteoporosis care gap in women: the Canadian Multicentre Osteoporosis Study.
Canadian women over 50 years old were studied over a 10-year period to see if those who sustained a fracture (caused by minimal trauma) were receiving the recommended osteoporosis therapy. We found that approximately half of these women were not being treated, indicating a significant care gap in osteoporosis treatment. Prevalent fragility fracture strongly predicts future fracture. Previous studies have indicated that women with fragility fractures are not receiving the indicated treatment. We aimed to describe post fracture care in Canadian women using a large, population-based prospective cohort that began in 1995-1997. We followed 5,566 women over 50 years of age from across Canada over a period of 10 years in the Canadian Multicentre Osteoporosis Study. Information on medication use and incident clinical fragility fractures was obtained during a yearly questionnaire or interview and fractures were confirmed by radiographic/medical reports. Over the 10-year study period, 42-56% of women with yearly incident clinical fragility fractures were not treated with an osteoporosis medication. During year 1 of the study, 22% of the women who had experienced a fragility fracture were on treatment with a bisphosphonate and 26% were on hormone therapy (HT). We were not able to differentiate HT use for menopause symptoms vs osteoporosis. Use of bisphosphonate therapy increased over time; odds ratio (OR) for use at year 10 compared to use at year 1 was 3.65 (95% confidence interval (CI) 1.83-7.26). In contrast, HT use declined, with an OR of 0.07 (95%CI 0.02-0.24) at year 10 compared to year 1 of the study. In a large population-based cohort study, we found a therapeutic care gap in women with osteoporosis and fragility fractures. Although bisphosphonate therapy usage improved over time, a substantial gap remains.


Prospective sensitivities of atom interferometers to gravitational waves and ultralight dark matter.
We survey the prospective sensitivities of terrestrial and space-borne atom interferometers to gravitational waves generated by cosmological and astrophysical sources, and to ultralight dark matter. We discuss the backgrounds from gravitational gradient noise in terrestrial detectors, and also binary pulsar and asteroid backgrounds in space-borne detectors. We compare the sensitivities of LIGO and LISA with those of the 100 m and 1 km stages of the AION terrestrial AI project, as well as two options for the proposed AEDGE AI space mission with cold atom clouds either inside or outside the spacecraft, considering as possible sources the mergers of black holes and neutron stars, supernovae, phase transitions in the early Universe, cosmic strings and quantum fluctuations in the early Universe that could have generated primordial black holes. We also review the capabilities of AION and AEDGE for detecting coherent waves of ultralight scalar dark matter. AION-REPORT/2021-04 KCL-PH-TH/2021-61, CERN-TH-2021-116 This article is part of the theme issue 'Quantum technologies in particle physics'.


Long-Term Outcomes after Minimally Invasive Aortic Valve Surgery through Right Anterior Minithoracotomy.
Background Minimally invasive aortic valve surgery (MIAV) through a right anterior minithoracotomy evolved to an accepted procedure with favorable short- and mid-term outcomes, whereas long-term results lack. The aim of this study was to evaluate the long-term outcomes. Materials and Methods All our MIAV patients were included (n = 225). Mean age was 68 ± 12 years, 29% were older than 75 years, and median EuroSCORE was 5 (0-11). Baseline characteristics, inhospital outcomes, and follow-up information about survival, major adverse cardiac and cerebrovascular events (MACCE), and need for reoperation were collected and analyzed. Results In this study, 30-day mortality was 1.3%, and there was no permanent stroke. Mean follow-up time was 69.65 ± 24 months, being the longest so far reported in the literature. At 1 and 7 years, survival was 95.8 and 79%, freedom from MACCE 98.1 and 95.7%, and from reoperation 99.5 and 98.7%, respectively. Conclusion MIAV is safe and feasible with favorable long-term outcomes. In the future, it could serve as benchmark for interventional methods as soon as indications are expanded to young and low-risk patients. Randomized studies are needed to compare the long-term outcomes of these approaches.


Differential Scanning Calorimetry Analyses of Idebenone-Loaded Solid Lipid Nanoparticles Interactions with a Model of Bio-Membrane: A Comparison with In Vitro Skin Permeation Data.
Differential scanning calorimetry (DSC) has emerged as a helpful technique both to characterize drug delivery systems and to study their interactions with bio-membranes. In this work, we compared idebenone (IDE)-loaded solid lipid nanoparticle (SLN) interactions with bio-membranes assessed by DSC with previous in vitro skin penetration data to evaluate the feasibility of predicting IDE skin penetration using DSC analyses. In vitro interactions experiments were performed using multi-lamellar liposomes as a model of bio-membrane. Enthalpy changes (ΔH) and transition temperature (Tm) were assessed during nine repeated DSC scans to evaluate IDE-loaded SLN⁻bio-membrane interactions over time. Analyzing ΔH and Tm values for each scan, we observed that the difference of ΔH and Tm values between the first and the last scan seemed to be related to SLN ability to locate IDE in the epidermis and in the stratum corneum, respectively. Therefore, the results of this study suggest the possibility of qualitatively predicting in vitro IDE skin penetration from IDE-loaded SLN utilizing the calorimetric parameters obtained from interaction experiments between the carriers under investigation and a model of bio-membrane.


ARNT PAS-B has a fragile native state structure with an alternative beta-sheet register nearby in sequence space.
The aryl hydrocarbon receptor nuclear translocator (ARNT) is a basic helix-loop-helix Period/ARNT/Single-minded (bHLH-PAS) protein that controls various biological pathways as part of dimeric transcriptional regulator complexes with other bHLH-PAS proteins. The two PAS domains within ARNT, PAS-A and PAS-B, are essential for the formation of these complexes because they mediate protein-protein interactions via residues located on their beta-sheet surfaces. While investigating the importance of residues in ARNT PAS-B involved in these interactions, we uncovered a point mutation (Y456T) on the solvent-exposed beta-sheet surface that allowed this domain to interconvert with a second, stable conformation. Although both conformations are present in equivalent quantities in the Y456T mutant, this can be shifted almost completely to either end point by additional mutations. A high-resolution solution structure of a mutant ARNT PAS-B domain stabilized in the new conformation revealed a 3-residue slip in register and accompanying inversion of the central Ibeta-strand. We have demonstrated that the new conformation has >100-fold lower in vitro affinity for its heterodimerization partner, hypoxia-inducible factor 2alpha PAS-B. We speculate that the pliability in beta-strand register is related to the flexibility required of ARNT to bind to several partners and, more broadly, to the abilities of some PAS domains to regulate their activities in response to small-molecule cofactors.


A new method to cross-link a polyplex for enhancing in vivo stability and transfection efficiency.
Disulfide-exchange was found to cross-link the polyplex of disulfide-containing poly(amido amine) and pDNA with heating of the polyplex solution over a short time. The cross-linked polyplexes based on disulfide-containing poly(amido amine) have excellent stability under physiological salt conditions, and have significantly enhanced transfection activity in the serum media compared to non-cross-linked polyplexes. In vivo, ICR mice were injected with the polyplex through the tail vein, the results show that the transfection efficiency of the cross-linked polyplex is higher than that of the non-cross-linked variety. Furthermore, the polyplex containing Cy5 labelled DNA was also injected into the mice to illustrate the stability and distribution of the polyplex, cross-linked polyplexes show a much brighter luminescence than the non-cross-linked ones. This method does not need a cross-linker or catalyst, and there are no impurities produced, it may be an elegant approach to resolve the dilemma of in vivo application of a DNA polyplex, with excellent stability whilst in circulation and a rapid unpacking of the polyplex inside the cells.


The Cultural Importance of Plants in Western African Religions.
This paper explores the importance of elements of the natural world, and particularly plants, among people of different religious affiliations in western Africa. Plants play an overriding role in African folk religions, which in turn are closely associated with health practices and influence management decisions concerning natural resources. In spite of the extensive literature documenting ritual plant use, the cultural importance of plants in this context has not been systematically assessed. Our objective was to see whether the importance of plants was reflected in people's conceptions of global (i.e., Christianity, Islam) and folk religions (i.e., Vodoun and Bwiti) in Benin (West Africa) and Gabon (Central Africa). By performing a cultural domain analysis (CDA) with 96 individuals, we found that, regardless of the religious affiliation of informants, plants and other elements of the natural world were more present in people's notions of folk religions than in global religions. We conclude by reflecting on the potentials and limitations of the data presented here as a starting point to explore the topic of cultural keystone species.


Owner observations regarding cat scratching behavior: an internet-based survey.
This study was performed to examine aspects of the cat, environment and scratching post that might influence scratching behavior, in an effort to determine how inappropriate scratching behavior might be refocused on acceptable targets. An internet survey, posted on several public websites, gathered details about scratching behavior, as described by owners in their home environments, from 4331 respondents over a 4 month period. Responses from 39 different countries were analyzed, mostly from the USA, Canada and the UK. Owners offered traditionally recommended scratching substrates including rope, cardboard, carpet and wood. Rope was most frequently used when offered, although carpet was offered most commonly. Most owners provided at least one scratching post; cats scratched the preferred substrate more often when the post was a simple upright type or a cat tree with two or more levels and at least 3 ft high. Narrower posts (base width ⩽3 ft) were used more often than wider posts (base width ⩾5 ft). Intact or neutered cats (males and females) were as likely to scratch inappropriately, and inappropriate scratching decreased with age. Geriatric cats between the ages of 10 and 14 years preferred carpet substrate most frequently; all other ages preferred rope first. Inappropriate scratching decreased as the different types/styles of posts increased in the home. Inappropriate scratching did not increase if the number of cats or dogs increased in the household. Declawed cats were preventatively declawed most often to prevent household item destruction. Although cats can have individual preferences, our data provide a starting point for veterinarians recommending scratching posts to clients.


Room temperature X-ray absorption spectroscopy of metalloenzymes with drop-on-demand sample delivery at XFELs.
X-ray crystallography and X-ray spectroscopy using X-ray free electron lasers plays an important role in understanding the interplay of structural changes in the protein and the chemical changes at the metal active site of metalloenzymes through their catalytic cycles. As a part of such an effort, we report here our recent development of methods for X-ray absorption spectroscopy (XAS) at XFELs to study dilute biological samples, available in limited volumes. Our prime target is Photosystem II (PS II), a multi subunit membrane protein complex, that catalyzes the light-driven water oxidation reaction at the Mn4CaO5 cluster. This is an ideal system to investigate how to control multi-electron/proton chemistry, using the flexibility of metal redox states, in coordination with the protein and the water network. We describe the method that we have developed to collect XAS data using PS II samples with a Mn concentration of <1 mM, using a drop-on-demand sample delivery method.


Inhibition and enhancement of Friend leukemia virus by pyran copolymer.
Inhibition or enhancement of Friend leukemia virus disease could be produced by treatment of mice with the immunopotentiator, pyran copolymer. The result depended on the route of inoculation of the drug. Prophylactic administration of the drug i.p. retarded splenomegaly, reduced splenic foci, and increased survival time of mice infected with Friend leukemia virus. Conversely, when the same dose and regimen of pyran was administered i.v., splenomegaly was enhanced, splenic foci were increased, and survival time was decreased. Histopathological examination of the spleens of mice revealed that i.p. pyran administration caused a marked increase in the splenic marginal zone with some increase in erythropoiesis in the red pulp, while i.v. pyran administration did not markedly change the splenic marginal zone but caused an early and sustained increase in erythropoiesis in the red pulp.


[Complex karyotype abnormalities in pediatric acute myeloid leukemia].
A majority of the data on the prognostic significance of distinct chromosome changes and combinations of them in pediatric acute myeloid leukemia (AML) has been derived from adult studies, with not numerous published data in pediatric patients. One of points needed to be clarified is prognostic significance of complex karyotype (at least 3 unrelated abnormalities). We investigated characteristic features of complex karyotype in newly diagnosed pediatric AML de novo. Cell clones with complex karyotype were found in 35 of 254 (13.8%) patients at the age from 0 to 15 years studied prior to therapy. The group was divided into 2 subgroups depending on presence of favorable chromosome abnormalities, i.e. (see symbols)(8;21), t(15;17) and inv(16). The abnormalities were absent in 20 cases (1st subgroup), in 15 remaining patients they were identified (2nd subgroup). In 2nd subgroup karyotypes were not so considerably changed and no adverse risk markers were detected as distinct from 1st subgroup. New data were obtained for complex karyotype differences of adult and pediatric AML. In the great majority (76%) of complex karyotypes in our adult patients chromosome abnormalities associated with adverse risk were found but in pediatric patients their frequency was significantly less (30%). The highest rate of complex karyotype we observed in children at the age from 0 to 3 years. Similar data were not published earlier. Complex karyotype is considered to be characteristic of older AML patients and in the majority of the patients the karyotype contains markers of adverse risk. Possibly, worse outcome in older AML patients is connected with the markers but not with multiple chromosome changes. New data of frequency and the peculiarities of complex karyotype in pediatric AML are important for understanding of AML pathogenesis and for development a more effective AML treatment.


Prevalence, location and symmetry of noncatastrophic ligamentous suspensory apparatus lesions in California Thoroughbred racehorses, and association of these lesions with catastrophic injuries.
Knowledge of the site distribution of ligamentous injuries facilitates clinical diagnosis of suspensory apparatus conditions. To determine if lesions within the suspensory ligament (SL) and distal ligaments of the proximal sesamoid bones (DSLs) were associated with suspensory apparatus failure or metacarpal lateral condylar fracture in California Thoroughbred racehorses. Cross-sectional study. Suspensory apparatus specimens from 327 deceased Thoroughbred racehorses were sectioned within the SL body and branches, and oblique and straight DSLs. Purple lesions ≥2 mm in width were categorised as moderate and paler or smaller lesions as mild. Associations between moderate lesions and age, sex, racetrack and cause of death were evaluated using multivariable logistic regression. Moderate lesions were evident in 16% and milder lesions in 77% of racehorses. Moderate lesions occurred with similar frequency in SL branches and oblique DSLs. Moderate lesions were more likely to occur in horses that died as a result of suspensory apparatus failure (odds ratio [OR] = 4.60; 95% confidence interval [CI] 1.61-13.13; and P = 0.004) or metacarpal lateral condylar fracture (OR = 5.05; 95% CI 1.42-17.93; and P = 0.012) compared with horses that died from nonmusculoskeletal causes, and in horses aged ≥7 years horses compared with 2-year-old horses (OR = 5.33; 95% CI 1.44-19.75; and P = 0.012). Moderate lesions are common in the SL branches and oblique DSLs of racehorses, and may be associated with risk for suspensory apparatus failure and metacarpal condylar fracture. Monitoring health of the suspensory apparatus ligamentous structures may be a simple means of assessing fatigue in, and preventing more extensive injuries to, the forelimb suspensory apparatus and metacarpal condyles.


Systemic immune inflammation index: a novel predictor for coronary collateral circulation.
Recently, a new inflammatory and prognostic marker has emerged called as Systemic Immune Inflammation Index (SII). In the current study, we searched the relation between SII and Coronary Collateral Circulation (CCC) formation in stable Coronary Artery Disease (CAD). 449 patients with stable CAD who underwent coronary angiography and documented coronary stenosis of 95% or more in at least one major coronary vessel were included in the study. The study patients were divided into two groups according to the Rentrop score as well CCC (Rentrop 2-3) and bad CCC (Rentrop 0-1). Blood samples for SII and other laboratory parameters were gathered from all the patients on admission. The SII score was formulized as platelet × neutrophil/lymphocyte counts. Patients, who had developed bad CCC had a higher C-reactive protein (CRP), neutrophil/lymphocyte ratio (NLR), platelets/lymphocyte ratio (PLR) and SII levels compared to those who had developed well CCC (p < 0.001, for all). Multivariate logistic regression analysis showed that high levels of SII was an independent predictor of bad CCC (OR: 1.005, 95% confidence interval (CI): 1.003-1.006, p < 0.001) together with dyslipidemia, high levels of CRP and NLR. In Receiver Operator Characteristic curve (ROC) analysis, the optimal cutoff value of SII to predict poor CCC was found to be 729.8, with 78.4% sensitivity and 74.6% specificity (area under ROC curve = 0.833 (95% CI: 0.777-0.889, p < 0.001). We have demonstrated that SII, a novel cardiovascular risk marker, might be used as one of the independent predictors of CCC development.


The in vitro assessment of anti-human lymphocyte globulin.
A simple reproducible in vitro test of anti-human lymphocyte globulin activity is discribed. The method is a "microtitre" complement-fixation test using human platelets as a source of antigen, guinea-pig complement and an amboceptor of sheep red blood cells sensitized by rabbit anti-sheep haemolysin.


We Offer Renal Replacement Therapy to Patients Who Are Not Benefitted by It.
Nephrologists offer renal replacement therapy (RRT) to patients who are unlikely to benefit in part because of our discomfort discussing goals of care in the setting of an uncertain prognosis for a given individual. Permanent neurological impairment, terminal illness (life expectancy <6 months), medical conditions precluding the safe delivery of dialysis, elderly patients with poor prognosis, and those who begin "early" RRT are categories of patients for whom dialysis may not be beneficial. Successful use of time-limited trials of dialysis may reduce the number of patients who are started on RRT without significant benefit. However, clear achievable milestones and goals need to be incorporated into plans for time-limited trials to ensure that continuing RRT beyond the trial period is appropriate. The lack of information on outcomes and symptom management using a "palliative approach" to dialysis suggests this should not be a clinical option until additional study is done and efficacy data available. Clinical practice guidelines are available to assist nephrologists in the appropriate withholding of RRT.


The impact of dedicated medication nurses on the medication administration error rate: a randomized controlled trial.
Concerns about hospital medication safety mount as the pace of new drug releases accelerates. We performed a randomized study at 2 hospitals (A and B) to examine whether the medication administration error rate could be decreased by having "dedicated" nurses focus exclusively on administering drugs. "Medication nurses," after receiving a brief review course on safe medication use, were responsible solely for drug delivery for up to 18 patients each. "General nurses," who did not attend the course, provided comprehensive care, including drug delivery, for 6 patients each. A direct observation technique was used to record drug errors, process-variation errors, and total errors. At both hospitals combined, the total error rate was 15.7% for medication nurses and 14.9% for general nurses (P<.84). Comparing hospitals, the total error rate for medication nurses at hospital B was significantly higher than it was at hospital A (19.7% vs 11.2%; P<.04). At hospital A, there was a significantly lower error rate for medication nurses than for general nurses in the surgical units (P<.01) but no significant differences in total errors comparing nurse types in the medical units (P<.77). This trial suggests that use of dedicated medication nurses does not reduce medication error rates. However, subgroup analysis indicates that medication nurses might be useful in some settings. The differences in findings at the 2 hospitals and their differences in medication-use processes reinforce the concept that medication errors are usually related to systems design issues.


Microvascularization and histomorphology of lateral line organs in adult Xenopus laevis.
The microvasculariaztion of the lateral line organs (LLOs) of the adult pipid frog, Xenopus laevis was studied by scanning electron microscopy of vascular corrosion casts (VCCs) and correlative light microscopy of paraplast embedded tissues sections. Scanning electron micrographs of VCCs revealed that each neuromast within the LLO rests on a distinct bowl-like capillary network (vascular bowl). One to three vascular bowls were supplied by an ascending arteriole and drained by a descending venule towards the skin deep dermal vascular network. Blood flow regulation mechanisms in form of intimal cushions were present at the origin of ascending arterioles supplying LLOs, microvenous valves were present at the confluence of deep dermal venules and veins. This together with sprouting and nonsprouting angiogenesis (intussusceptive microvascular growth) found in vascular bowls demonstrate that in adult Xenopus the capillary bed of LLO's still can be adjusted to changing energetic needs.


The effect of cartilage dehydration and rehydration on quantitative ultrashort echo time biomarkers.
The effect of dehydration of ex vivo cartilage samples and rehydration with native synovial fluid or normal saline on quantitative ultrashort echo time (UTE) biomarkers are unknown. We aimed to investigate the effect of cartilage dehydration-rehydration on UTE biomarkers and to compare the rehydration capabilities of native synovial fluid and normal saline. A total of 37 cartilage samples were harvested from patients (n=5) who underwent total knee replacement. Fresh cartilage samples were exposed to air to dehydrate for 2 hours after baseline magnetic resonance (MR) scanning, then randomly divided into two groups: one soaking in native synovial fluid (n=17) and the other in normal saline (n=20) to rehydrate for 4 hours. UTE-based biomarkers [T1, adiabatic T1r (AdiabT1r), macromolecular fraction (MMF), magnetization transfer ratio (MTR), and T2*] and sample weights were evaluated for fresh, dehydrated, and rehydrated cartilage samples. Differences and agreements between groups were assessed using the values of fresh cartilage samples as reference standard. Dehydrating in air for 2 hours resulted in significant weight loss (P=0.000). T1, AdiabT1r, and T2* decreased significantly while MMF and MTR increased significantly (all P<0.02). Non-significant differences were observed in cartilage weights after rehydrating in both synovial fluid and normal saline, with P values being 0.204 and 0.769, respectively. There were no significant differences in T1, AdiabT1r, MMF, and MTR after rehydrating in synovial fluid (P>0.0167, with Bonferroni correction) while T2* (P=0.001) still had significant differences compared with fresh samples. However, no significant differences were detected for any of the evaluated UTE biomarkers after rehydrating in normal saline (all P>0.05). No differences were detected in the agreement of UTE biomarker measurements between fresh samples and samples rehydrated with synovial fluid and normal saline. Cartilage dehydration resulted in significant changes in UTE biomarkers. Rehydrating with synovial fluid or normal saline had non-significant effect on all the evaluated UTE biomarkers except T2* values, which still had significant differences compared with fresh samples after rehydrating with synovial fluid. No significant difference was observed in the rehydration capabilities of native synovial fluid and normal saline.


[Clinical and neurobiological aspects of long-term administration of psychotropic drugs].
In neuroleptic therapy for psychotic illnesses, clinical improvement occurs much later than central dopamine blockade, and its time course varies widely among patients. A hypothesis explaining neuroleptic-responsive illness cannot be explained by dopamine blockade alone. Nevertheless, experimental data suggest that this mechanism may be a step in the therapeutic process for schizophrenia. Explanations are suggested for the time lag in therapeutic response for neuroleptics, including the hypothesis of delayed inactivation of mid-brain dopamine neurones. Chronic benzodiazepine treatment elicits adaptive responses in the CNS that are manifested as functional tolerance and physical dependence. Possible mechanisms involved in such a profound alteration of neuronal functioning are suggested. Down regulation of benzodiazepine receptors has been shown to be related to functional tolerance under certain conditions. The effect of repeated treatment with antidepressants is compatible with the hypothesis that changes in central monoamine transmission are involved in the activity of these drugs. Beta-adrenergic receptors are desensitized and their density is decreased; alpha-2 adrenoreceptors sensitivity is reduced, and post-synaptic serotoninergic receptors sensitivity is increased. It remains to be clarified whether some of the changes have larger role than others or whether they all contribute to the psychotropic drug activity in the therapeutic process.


Bridging patients with hepatocellular cancer waiting for liver transplant: all the patients are the same?
Liver transplant (LT) is considered the best curative treatment for patients with cirrhosis and hepatocellular carcinoma (HCC) within Milan criteria. The possibility to perform LT in HCC patients is limited by the liver grafts supply; indeed, the shortage of donors often leads to a long time on waiting list and then to dropout because of tumor progression. Bridging therapies are neo-adjuvant treatments given to patients on LT waitlist, with the aim to prevent tumor progression and to reduce dropout rate. Many bridging modalities have been proposed. The choice of each treatment is based on the characteristics of the patient, liver function, comorbidities and on the number, dimensions and localization of HCC. This review article describes several types of bridging therapies, focusing on the indications for different kind of patients.


Multivitamin mineral supplementation in patients with chronic fatigue syndrome.
Chronic fatigue syndrome (CFS) is characterized by medically unexplained persistent or reoccurring fatigue lasting at least 6 months. CFS has a multifactorial pathogenesis in which oxidative stress (OS) plays a prominent role. Treatment is with a vitamin and mineral supplement, but this therapeutic option so far has not been properly researched. This prospective study included 38 women of reproductive age consecutively diagnosed by CDC definition of CFS and treated with a multivitamin mineral supplement. Before and after the 2-month supplementation, SOD activity was determined and patients self-assessed their improvement in 2 questionnaires: the Fibro Fatigue Scale (FFS) and the Quality of Life Scale (SF36). Results There was a significant improvement in SOD activity levels; and significant decreases in fatigue (p=0.0009), sleep disorders (p=0.008), autonomic nervous system symptoms (p=0.018), frequency and intensity of headaches (p=0.0001), and subjective feeling of infection (p=0.0002). No positive effect on quality of life was found. Treatment with a vitamin and mineral supplement could be a safe and easy way to improve symptoms and quality of life in patients with CFS.


The impact of the Consumer Protection Act on pharmacists.
The Consumer Protection Act of 2008 has had far-reaching consequences for suppliers of goods and services in South Africa. The implementation of the Act has important implications for all suppliers who enter into 'consumer transactions.' This article aims to stimulate awareness of the legal consequences of the Act arising from day-to-day situations occurring in the pharmacy, and to highlight the compliance obligations that the Act creates for pharmacists.


The dissociation of exocytosis and respiratory stimulation in leucocytes by ionophores.
By exploiting the unique characteristics of three ionophores, experimental conditions were found which permit the dissociation of respiratory stimulation from secretion in polymorphonuclear leucocytes. A marked stimulation of respiration was produced by ionophore X537A, which binds and transports both alkali-earth and alkali cations. The stimulatory activity of this ionophore was the same at either high or low Na+/K+ ratios in the medium and was virtually unaffected by extracellular Ca2+. A slight stimulation of oxygen consumption was also caused by the K+-selective ionophore valinomycin and by ionophore A23187, which complexes and transfers bivalent cations. Ionophore X537A and valinomycin were unable to stimulate selective release of granuleassociated beta-glucuronidase and gradually increased cell fragility, as monitored by increased leakage of lactate dehydrogenase. Ionophore A23187 slightly increased exocytosis of beta-glucuronidase. In a Mg2+-free medium, Ca2+, added simultaneously with ionophore A23187, greatly enhanced respiration and secretion of the granule enzyme. If Ca2+ was added a few minutes after the ionophore, exocytosis occurred, but no respiratory burst was observed. If the latter experiment was repeated in the presence of extracellular Mg2+, both secretion and respiration were stimulated. This effect was not produced by Mn2+ or Ba2+. It is proposed that Ca2+ is required for triggering selective secretion of granule enzymes from leucocytes is caused by an intracellular redistribution of cations, which may invovle Mg2+-dependent mechanisms.


Autonomic mechanisms underlying capsaicin induced oral sensations and salivation in man.
The effects of capsaicin, citric acid and nicotine applied to the apex or radix of the tongue on taste sensations and salivation were studied in relation to the presence of substance P immunoreactive neurones in man. Application of capsaicin (30 micron) to the apex of the tongue or to the palatinal mucosa, but not to the radix of the tongue, caused a reproducible burning sensation and salivation from the submandibular-sublingual and parotid glands. The salivation response to capsaicin was reduced by methylscopolamine pretreatment. Similar levels of substance P immunoreactivity were present in the lingual apex and radix area (including vallate papillae) of man, while in the cat about 4 times higher levels of substance P immunoreactivity were present in the vallate papillae than in the lingual apex. Immunohistochemistry showed that in the cat many substance P immunoreactive nerves were associated with the taste buds of the vallate papillae, while in man substance P immunoreactive fibres were only seen penetrating into the epithelium of the lingual apex. In addition some subepithelial blood vessels in all regions were surrounded by substance P immunoreactive nerves in both cat and man. Citric acid application to the tongue apex caused both submandibular-sublingual and parotid salivary secretion concomitant with a burning sensation. Salivary secretion was also seen after citric acid application to the radix of the tongue. This response was associated with a sour taste. The salivation response to citric acid was not significantly reduced by methylscopolamine pretreatment. Lingual apex application of nicotine was associated with a sweet taste and a small rise in salivary secretion rate. This response was not significantly reduced by methylscopolamine. In conclusion, the sensitivity to capsaicin of the human tongue is restricted to the apex portion. This is in parallel with the occurrence of intraepithelial substance P immunoreactive nerve fibres. Capsaicin induced salivary secretion seems mainly to be mediated via parasympathetic, cholinergic reflex mechanisms. Citric acid and nicotine induced salivation responses are comparatively more resistant to methylscopolamine pretreatment.


Associations between nonverbal behaviors and subsequent sexual attitudes and behaviors of sexually abused and comparison girls.
This prospective, longitudinal study examined a sample of sexually abused and comparison girls to determine (a) whether there were patterns of behavior that differed between the groups and (b) whether nonverbal behaviors assessed at the initial visit (n = 147; M = 11.11 years; SD = 3.02) might predict sexual attitudes and behaviors at a later point in development (n = 144; M = 18.52 years; SD = 3.52). At the initial assessment, nonverbal behaviors during an interaction with an unknown male interviewer were factor analyzed revealing 3 factors: wary (e.g., pouting), affiliative (e.g., chin resting on hand), and coy (e.g., tongue show). Abused girls scored higher on the coy factor that was related to earlier age at first voluntary intercourse later in development (approximately 7 years later). High scores on the affiliative factor were related to higher sexual permissiveness and less negative attitudes toward sex. Results indicate that sexually abused girls showed early maladaptive patterns in interpersonal interactions, which were subsequently related to risky sexual attitudes and behaviors.


Cerebral autoregulation in children during sevoflurane anaesthesia.
Little is known about cerebral autoregulation in children. The aim of this study was to examine cerebral autoregulation in children. Cerebral autoregulation testing was performed during less than 1 MAC sevoflurane anaesthesia in children (from 6 months to 14 yr) and in adults (18-41 yr). Mean middle cerebral artery flow velocities (V(MCA)) were measured using transcranial Doppler ultrasonography. Mean arterial pressure (MAP) was increased to whichever was greater: 20% above baseline or (i) 80 mm Hg for less than 9 yr, (ii) 90 mm Hg for 9-14 yr, and (iii) 100 mm Hg for adults. Cerebral autoregulation was considered intact if the autoregulatory index was > or =0.4. There were 13 subjects less than 2 yr old (Group 1), 13 subjects 2-5 yr (Group II), 14 subjects 6-9 yr (Group III), 12 subjects 10-14 yr (Group IV), and 12 adults (Group V; control group). All subjects had an autoregulatory index > or =0.4. There was no difference in autoregulatory index between children in Groups I-IV or between children and adults. We found no age-related differences in autoregulatory capacity during low-dose sevoflurane anaesthesia. We report no differences in autoregulatory capacity between children and adults.


Autonomic Symptoms Correlate with Non-Autonomic Non-Motor Symptoms and Sleep Problems in Patients with Parkinson's Disease.
The objective of this study was to clarify the relationship between autonomic and non-autonomic non-motor symptoms in patients with Parkinson's disease (PD). Sixty-five PD patients were included in this study (27 men and 38 women; aged 68.5 ± 10.0; Hoehn and Yahr (HY) stage 2.6 ± 1.1). The autonomic symptoms were evaluated by the Japanese version of the Scales for outcomes in PD autonomic (SCOPA-AUT) questionnaire. The patients were assessed with the mini-mental state examination (MMSE), PD sleep evaluation scale-2 (PDSS-2), Epworth sleepiness scale (ESS) and Beck's depression inventory II (BDI-II). The Non-Motor Symptom Scale (NMSS) total scores and subscores of non-autonomic non-motor symptom domains (sleep/fatigue, mood/cognition, perceptual problems/hallucination, and attention/memory) were evaluated. A dopamine transporter (DAT) scan, metaiodobenzylguanidine (MIBG) myocardial scintigraphy, and card type olfactory identification test (open essence [OE, Wako]) were performed. The SCOPA-AUT total score was positively correlated with the disease duration, HY stage, levodopa equivalent dose, PDSS-2, ESS, BDI-II and non-autonomic NMSS and inversely correlated with the MMSE. The high-SCOPA-AUT group (≥9) had lower MMSE scores and higher PDSS-2, ESS, BDI-II and non-motor NMSS scores than the low-SCOPA-AUT group (< 9). The DAT scan, MIBG uptake and OE score did not differ between the groups. In a stepwise linear regression analysis, which excluded possibly overlapping items among the scales, the subtotals of PDSS-2 items, except for item 8 (nocturia), (p < 0.0001) and non-autonomic NMSS domains (p = 0.00040) were significant predictors of the total SCOPA-AUT score. Our study shows significant correlations among autonomic symptoms, PD-related sleep problems and non-autonomic non-motor symptoms in PD patients.


Protein and calorie intake, nitrogen losses, and nitrogen balance in children undergoing chronic peritoneal dialysis.
We performed 22 nitrogen balance (NB) studies of three days' duration in 19 children (8.7 +/- 3.8 years) on chronic peritoneal dialysis (CPD) for 19.4 +/- 16.4 months. The dietary intakes were assessed by the double weighing method. Total nitrogen, protein, urea, and creatinine were analyzed in the dialysate and urine collected daily. Total nitrogen was also determined in the feces collected over the whole NB study period, using vegetable charcoal as a marker. The protein intake was 1.64 +/- 0.50 g/kg/day, corresponding to 126 +/- 40% of the recommended daily allowance (RDA) for normal children of the same age, and the calorie intake (diet+glucose from dialysate) reached 75 +/- 26% of RDA. Nitrogen losses were: 0.177 +/- 0.052 g/kg/day with peritoneal fluid and urine, and 0.028 +/- 0.018 g/kg/day with feces. The NB, positive in 17 out of 22 studies, ranged from -116 to +167 mg/kg/day (mean 44.0 +/- 66.2 mg/kg/day). A direct and significant correlation between NB and nitrogen intake (g/kg/day) (r = 0.562, p < 0.05) and total calorie intake (cal/kg/day) (r = 0.483, p < 0.05) has been observed. These data confirm the need to ensure in children on CPD an adequate nutritional intake, and further support the efforts to improve calorie intake.


Comprehensive Analysis of AR Alterations in Circulating Tumor DNA from Patients with Advanced Prostate Cancer.
Somatic alterations in circulating tumor DNA (ctDNA) may be associated with treatment response or prognosis in prostate cancer (PCa). The goal was to characterize androgen receptor gene (AR) amplifications and mutations detected in ctDNA from patients with PCa and to further understand the somatic genetic heterogeneity of advanced prostate cancer. This study included a heterogeneous group of 892 patients with advanced PCa (predominantly castrate-resistant prostate cancer) with AR alterations detected in ctDNA that underwent next-generation sequencing of 54 to 73 genes via Guardant360 testing (Guardant Health, Inc., Redwood City, CA). Distribution and summary of AR alterations detected, the association of AR alterations with other genes, and a pathway analysis are reported. The median absolute plasma copy number of AR amplifications was 3.3 (range, 1.2-165.2). Many patients had multiple AR mutations; a total of 112 unique mutations were identified in AR, including L702H (25%), T878A (14%), H875Y (11%), W742C (8%), W742L (4%), F877L (2%), and T878S (2%). Other ctDNA gene alterations in the Guardant assays included TP53 (50%), MYC (34%), BRAF (32%), PIK3CA (29%), MET (25%), CDK6 (26%), EGFR (24%), FGFR1 (21%), and APC (12%). Many of these non-AR alterations are not tissue verified in other studies. AR amplification cosegregated with alterations in MYC (p < .001), BRAF (p < .001), PIK3CA (p < .001), MET (p < .001), CDK6 (p < .001), EGFR (p < .001), FGFR1 (p = .391), and more. Alterations in APC were significantly associated with mutations in AR (p < .001). Several AR alterations and concomitant non-AR alterations that associate with drug resistance were detected. These findings provide additional insights into the heterogeneity of advanced prostate cancer. The goal was to characterize androgen receptor gene (AR) amplifications and mutations detected in circulating tumor DNA (ctDNA) from patients with prostate cancer in relation to non-AR gene alterations detected in the ctDNA landscape. The study included 892 patients with prostate cancer with AR alterations in ctDNA. AR alterations were significantly associated with other gene alterations detected in ctDNA. The common AR mutations found are linked to resistance to abiraterone, enzalutamide, or bicalutamide. Characterization of the circulating AR landscape and gene alterations provides potential additional insight into the somatic genetic heterogeneity of advanced prostate cancer.


NULISA: a proteomic liquid biopsy platform with attomolar sensitivity and high multiplexing.
The blood proteome holds great promise for precision medicine but poses substantial challenges due to the low abundance of most plasma proteins and the vast dynamic range of the plasma proteome. Here we address these challenges with NUcleic acid Linked Immuno-Sandwich Assay (NULISA™), which improves the sensitivity of traditional proximity ligation assays by ~10,000-fold to attomolar level, by suppressing assay background via a dual capture and release mechanism built into oligonucleotide-conjugated antibodies. Highly multiplexed quantification of both low- and high-abundance proteins spanning a wide dynamic range is achieved by attenuating signals from abundant targets with unconjugated antibodies and next-generation sequencing of barcoded reporter DNA. A 200-plex NULISA containing 124 cytokines and chemokines and other proteins demonstrates superior sensitivity to a proximity extension assay in detecting biologically important low-abundance biomarkers in patients with autoimmune diseases and COVID-19. Fully automated NULISA makes broad and in-depth proteomic analysis easily accessible for research and diagnostic applications.