The Maine Lumbar Spine Study, Part III. 1-year outcomes of surgical and nonsurgical management of lumbar spinal stenosis.
A prospective cohort study of patients with lumbar spinal stenosis recruited from the practices of orthopedic surgeons and neurosurgeons throughout Maine. To assess 1-year outcomes of patients with lumbar spinal stenosis treated surgically or nonsurgically. No randomized trials and few nonexperimental studies have compared surgical and nonsurgical treatment of patients with lumbar spinal stenosis. The authors' goal was to assess 1-year outcomes of patients with lumbar spinal stenosis treated surgically or nonsurgically. Eligible, consenting patients participated in baseline interviews and were then mailed follow-up questionnaires at 3, 6, and 12 months. Clinical data were obtained from a physician questionnaire. Outcomes included patient-reported symptoms of leg and back pain, functional status, disability, and satisfaction with care. One hundred forty-eight patients with lumbar spinal stenosis were enrolled, of whom 81 were treated surgically and 67 treated nonsurgically. On average, patients in the surgical group had more severe imaging findings and symptoms and worse functional status than patients in the nonsurgical group at entry. Few patients with mild symptoms were treated surgically, and few patients with severe symptoms were treated nonsurgically. However, of the patients with moderate symptoms, a similar percent were treated surgically or nonsurgically. One year after study entry, 28% of nonsurgically and 55% of surgically treated patients reported definite improvement in their predominant symptoms (P = 0.003). For patients with moderate symptoms, outcomes for surgically treated patients were also improved compared with those of nonsurgically treated patients. Surgical treatment remained a significant determinant of 1-year outcome, even after adjustment for differences between treatment groups at entry (P = 0.05). The maximal benefit of surgery was observed by the time of the first follow-up evaluation, which was at 3 months. Although few nonsurgically treated patients experienced a worsening of their condition, there was little improvement in symptoms and functional status compared with study entry. At a 1-year evaluation of patient-reported outcomes, patients with severe lumbar spinal stenosis who were treated surgically had greater improvement than patients treated nonsurgically. Comparisons of outcomes by treatment received must be made cautiously because of differences in baseline characteristics. A determination of whether the outcomes observed persist requires long-term follow-up.


Involving Caregivers of People With Dementia to Validate Booklets on Food-Related Activities: A Qualitative Think-Aloud Study.
This study is the first to explore informal dementia caregivers' perceptions and outlook on written materials about all food-related processes: shopping, food preparation, and eating. The aim of the study was to develop and evaluate the content, format, and usefulness of two separate booklets (one newly developed and one existing) on food-related processes. Twenty dementia caregivers were provided with one of the two booklets, and a Think-Aloud method was used to gather information about their views on the booklets. The findings indicated that incorporating all three food processes in a new booklet could be beneficial for the participant. Shopping, purchasing food, driving, and dangers in the kitchen were addressed only in the developed booklet, and participants regarded them as important and useful areas to address. Therefore, this study has shown that tailored information may enhance caregivers' confidence and support them in making decisions to help them adapt to food-related changes.


Matrix-Assisted Ionization-Ion Mobility Spectrometry-Mass Spectrometry: Selective Analysis of a Europium-PEG Complex in a Crude Mixture.
The analytical utility of a new and simple to use ionization method, matrix-assisted ionization (MAI), coupled with ion mobility spectrometry (IMS) and mass spectrometry (MS) is used to characterize a 2-armed europium(III)-containing poly(ethylene glycol) (Eu-PEG) complex directly from a crude sample. MAI was used with the matrix 1,2-dicyanobenzene, which affords low chemical background relative to matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization (ESI). MAI provides high ion abundance of desired products in comparison to ESI and MALDI. Inductively coupled plasma-MS measurements were used to estimate a maximum of 10% of the crude sample by mass was the 2-arm Eu-PEG complex, supporting evidence of selective ionization of Eu-PEG complexes using the new MAI matrix, 1,2-dicyanobenzene. Multiply charged ions formed in MAI enhance the IMS gas-phase separation, especially relative to the singly charged ions observed with MALDI. Individual components are cleanly separated and readily identified, allowing characterization of the 2-arm Eu-PEG conjugate from a mixture of the 1-arm Eu-PEG complex and unreacted starting materials. Size-exclusion chromatography, liquid chromatography at critical conditions, MALDI-MS, ESI-MS, and ESI-IMS-MS had difficulties with this analysis, or failed. Graphical Abstract .


Importance of DNA stiffness in protein-DNA binding specificity.
From the first high-resolution structure of a repressor bound specifically to its DNA recognition sequence it has been shown that the phage 434 repressor protein binds as a dimer to the helix. Tight, local interactions are made at the ends of the binding site, causing the central four base pairs (bp) to become bent and overtwisted. The centre of the operator is not in contact with protein but repressor binding affinity can be reduced at least 50-fold in response to a sequence change there. This observation might be explained should the structure of the intervening DNA segment vary with its sequence, or if DNA at the centre of the operator resists the torsional and bending deformation necessary for complex formation in a sequence dependent fashion. We have considered the second hypothesis by demonstrating that DNA stiffness is sequence dependent. A method is formulated for calculating the stiffness of any particular DNA sequence, and we show that this predicted relationship between sequence and stiffness can explain the repressor binding data in a quantitative manner. We propose that the elastic properties of DNA may be of general importance to an understanding of protein-DNA binding specificity.


Spatial variation and inconsistency between estimates of onset of muscle activation from EMG and ultrasound.
Delayed onset of muscle activation can be a descriptor of impaired motor control. Activation onset can be estimated from electromyography (EMG)-registered muscle excitation and from ultrasound-registered muscle motion, which enables non-invasive measurements in deep muscles. However, in voluntary activation, EMG- and ultrasound-detected activation onsets may not correspond. To evaluate this, ten healthy men performed isometric elbow flexion at 20% to 70% of their maximal force. Utilising a multi-channel electrode transparent to ultrasound, EMG and M(otion)-mode ultrasound were recorded simultaneously over the biceps brachii muscle. The time intervals between automated and visually estimated activation onsets were correlated with the regional variation of EMG and muscle motion onset, contraction level and speed. Automated and visual onsets indicated variable time intervals between EMG- and motion onset, median (interquartile range) 96 (121) ms and 48 (72) ms, respectively. In 17% (computed analysis) or 23% (visual analysis) of trials, motion onset was detected before local EMG onset. Multi-channel EMG and M-mode ultrasound revealed regional differences in activation onset, which decreased with higher contraction speed (Spearman ρ ≥ 0.45, P < 0.001). In voluntary activation the heterogeneous motor unit recruitment together with immediate motion transmission may explain the high variation of the time intervals between local EMG- and ultrasound-detected activation onset.


Discovering Variable-Length Patterns in Protein Sequences for Protein-Protein Interaction Prediction.
To predict Protein-Protein Interactions (PPIs), there have recently been some attempts to use computational approaches and among them, sequence-based approaches are often preferred over other kinds of approaches as they do not require prior knowledge about proteins to perform their tasks. However, in deciding if two proteins may interact with each other, existing sequence-based approaches consider only fixed-length segments. We believe that if segments of variable-length can also be considered, interactions between proteins can be more accurately predicted. To consider variable-length segments for PPI predictions, we have developed a VLASPD algorithm. Given a database of protein sequences, VLASPD performs its tasks in several steps. The protein database is first searched to identify frequent sequence segments (FSSs) of different length. The different combinations of the presence and absence of these FSSs are then used to form different associative sequential patterns (ASPs). Based on a statistical measure, the ASPs that occur significantly frequently among proteins in the training set are then identified as significant associative sequential patterns (SASPs). If an SASP is found in a protein pair, it can be considered as providing some evidence to support or refute the existence of an interaction relationship between the protein pairs. The amount of evidence provided are then quantified with an information theoretic measure. How likely two proteins may interact with each other are then decided by the total amount of evidence provided by the SASPs found in the protein pairs. To test the effectiveness of VLASPD, we used several sets of real data. The experimental results show that VLASPD can be a promising approach for PPI prediction. The VLASPD is made available for use and testing at http://www.comp.polyu.edu.hk/~cslhu/resources/vlaspd/.


Hemorrhagic presentations of COVID-19: Risk factors for mortality.
We aim to characterize the incidence, risk for mortality, and identify risk factors for mortality in patients presenting with hemorrhage and COVID-19. This retrospective cohort study included a cohort of patients admitted to one of three major hospitals of our healthcare network including, an academic medical center and comprehensive stroke center, which accepts transfers for complex cases from eight community hospitals, during March 1 to May 1, 2020. All patients that received imaging of the neuroaxis and had positive PCR testing for COVID-19 were identified and reviewed by an attending neuroradiologist. Demographics and comorbidities were recorded. Biomarkers were recorded from the day of the hemorrhagic event. Vital signs from the day of the hemorrhagic event mechanical ventilation orders at admission were recorded. Imaging findings were divided into 5 subtypes; acute subdural hematoma (SDH), subarachnoid hemorrhage (SAH), multi-compartmental hemorrhage (MCH), multi-focal intracerebral hemorrhage (MFH), and focal intracerebral hemorrhage (fICH). Outcomes were recorded as non-routine discharge and mortality. We found a total of 35 out of 5227 patients with COVID-19 that had hemorrhage of some kind. Mortality for the entire cohort was 45.7 % (n = 16). SDH patients had a mortality rate of 35.3 % (n = 6), SAH had a mortality of 50 % (n = 1), MCH patients had a mortality of 71.4 % (n = 5), MFH patients had a mortality of 50 % (n = 2), fICH patients had a mortality of 40 % (n = 2). Patients with severe pulmonary COVID requiring mechanical ventilation (OR 10.24 [.43-243.12] p = 0.015), with INR > 1.2 on the day of the hemorrhagic event (OR 14.36 [1.69-122.14] p = 0.015], and patients presenting with spontaneous vs. traumatic hemorrhage (OR 6.11 [.31-118.89] p = 0.023) had significantly higher risk for mortality. Hemorrhagic presentations with COVID-19 are a rare but serious way in which the illness can manifest. It is important for neurosurgeons to realize that patients can present with these findings without primary pulmonary symptoms, and that severe pulmonary symptoms, elevated INR, and spontaneous hemorrhagic presentations is associated with increased risk for mortality.


Improved survival with recent Post-Transplant Lymphoproliferative Disorder (PTLD) in children with kidney transplants.
Post-transplant lymphoproliferative disorder (PTLD) has been associated with high mortality, but recent anecdotal survival appeared better. From 1988 to 2010, the NAPRTCS registry had 235 registered PTLD cases. We sent a special 25-point questionnaire study to the NAPRTCS centers with the most recent 150 cases to obtain additional follow-up data not collected in the master registry, our objective being to determine the recent outcomes after PTLD and determine prognostic factors. We received 92 completed responses, in which only 12 (13%) deaths were reported, 2 from nonmedical causes, 10 with a functioning graft. Kaplan-Meier-calculated patient survival was 90.6% at 1 year and 87.4% at 3, 4 and 5 years post-PTLD. Graft survival post-PTLD was 81.8% at 1 year, 68.0% at 3 years and 65.0% at 5 years. Seven patients received a retransplant after PTLD, with no PTLD recurrence reported. Using all 235 PTLD cases, the covariates associated with better patient survival were more recent year of PTLD diagnosis (adjusted hazard ratio AHR 0.86, p < 0.001), and with worse survival were late PTLD (AHR 1.98, p = 0.0176) and patient age above 13 at PTLD (AHR 3.43, p value 0.022). In children with kidney transplants, patient survival has improved with more recent PTLDs.


Cardiac resynchronization therapy: left or left-and-right for optimal symptomatic effect--the LOLA ROSE study.
Biventricular (BiV) pacing and left univentricular (LUV) pacing can each produce clinical benefits in heart failure. The impact of modern refinements in pacing optimization on the relative benefits of these two modes is unknown. We aimed to compare these two modes in patients with heart failure, using Echo-based optimization of each pacing mode. Paired data were collected on 18 patients (age 72 +/- 8 years; 16 male) with refractory heart failure symptoms, sinus rhythm, and LBBB with QRS duration>120 ms. Patients were randomized to an initial 8 weeks of either BiV or LUV pacing, followed by 8 weeks of the other mode, in a blinded cross-over design. Echocardiography was used to optimize atrioventricular delay for both modes and right ventricular-left ventricular offset for BiV mode. Peak oxygen consumption (baseline 13.6 +/- 2.7; BiV 15.8 +/- 3.0; LUV 15.2 +/- 3.1 mL/kg/min), 6 min walk distance (baseline 258 +/- 47; BiV 290 +/- 63; LUV 287 +/- 69 m), and scores on SF36 health questionnaire (baseline 41.5 +/- 16.8; BiV 58.6 +/- 19.6; LUV 51.8 +/- 21.3) did not differ between BiV and LUV modes. New York Heart Association class was significantly better in BiV than in LUV mode (P < 0.01). In this pilot study, we found no differences in major clinical outcome measures between the two modes of resynchronization.


Mapping considerations for optimal binary correlation filters.
The optimality of correlation filters is an important issue in applications of pattern recognition. We consider here both binary phase-only filters (BPOFs) and amplitude encoded binary phase-only filters (AE BPOFs) and study the results of optimizing the filters for a real world object (the Space Shuttle). We find that while only small improvements result from optimizing a BPOF, optimization of the AE BPOF is quite important in obtaining a useful correlation function. In the case of an AE BPOF, both signal-to-noise and peak-to-sidelobe measures must be studied. Computer simulation and experimental correlation results are presented.


Open channel deterministic lateral displacement for particle and cell sorting.
We present the use of capillary driven flow over patterned surfaces to achieve cheap and simple, but powerful separation of biologically relevant particle systems. The wide use of microfluidics is often hampered by the propensity for devices to clog due to the small channel sizes and the inability to access the interior of devices for cleaning. Often the devices can only be used for a limited duration and most frequently only once. In addition the cost and power requirements of flow control equipment limits the wider spread of the devices. We address these issues by presenting a simple particle- and cell-sorting scheme based on controlled fluid flow on a patterned surface. The open architecture makes it highly robust and easy to use. If clogging occurs it is straightforward to rinse the device and reuse it. Instead of external mechanical pumps, paper is used as a capillary pump. The different fractions are deposited in the paper and can subsequently be handled independently by simply cutting the paper for downstream processing and analyses. The sorting, based on deterministic lateral displacement, performs equivalently well in comparison with standard covered devices. We demonstrate successful separation of cancer cells and parasites from blood with good viability and with relevance for diagnostics and sample preparation. Sorting a mixture of soil and blood, we show the potential for forensic applications.


Stress and immunity in humans: a meta-analytic review.
This article presents a meta-analysis of the literature on stress and immunity in humans. The primary analyses include all relevant studies irrespective of the measure or manipulation of stress. The results of these analyses show substantial evidence for a relation between stress and decreases in functional immune measures (proliferative response to mitogens and natural killer cell activity). Stress is also related to numbers and percent of circulating white blood cells, immunoglobulin levels, and antibody titers to herpesviruses. Subsequent analyses suggest that objective stressful events are related to larger immune changes than subjective self-reports of stress, that immune response varies with stressor duration, and that interpersonal events are related to different immune outcomes than nonsocial events. We discuss the way neuroendocrine mechanisms and health practices might explain immune alteration following stress, and outline issues that need to be investigated in this area.


Morphological Plant Modeling: Unleashing Geometric and Topological Potential within the Plant Sciences.
The geometries and topologies of leaves, flowers, roots, shoots, and their arrangements have fascinated plant biologists and mathematicians alike. As such, plant morphology is inherently mathematical in that it describes plant form and architecture with geometrical and topological techniques. Gaining an understanding of how to modify plant morphology, through molecular biology and breeding, aided by a mathematical perspective, is critical to improving agriculture, and the monitoring of ecosystems is vital to modeling a future with fewer natural resources. In this white paper, we begin with an overview in quantifying the form of plants and mathematical models of patterning in plants. We then explore the fundamental challenges that remain unanswered concerning plant morphology, from the barriers preventing the prediction of phenotype from genotype to modeling the movement of leaves in air streams. We end with a discussion concerning the education of plant morphology synthesizing biological and mathematical approaches and ways to facilitate research advances through outreach, cross-disciplinary training, and open science. Unleashing the potential of geometric and topological approaches in the plant sciences promises to transform our understanding of both plants and mathematics.


Astronomy. Laser telemetry from space.
Space missions currently on the drawing boards are expected to gather data at rates exceeding the transmission capabilities of today's telemetry systems by many orders of magnitude. Even on current missions, onboard data compression techniques are being implemented to compensate for lack of transmission speed. But while data compression can minimize the loss of data, it is no substitute for transmitting all of the data through a faster communications link. The transmission problem will soon reach crisis proportions and will affect astronomical, Earth resources, geophysical, meteorological, planetary and other space science missions. To overcome this communications bottleneck, the authors advocate the implementation of telemetry systems based on near-infrared laser transmission techniques. The fiber-optics communications industry has developed most of the basic components required for signal transmission in this wavelength band, which should make such a system affordable on scales relevant to the cost of anticipated space science missions.


Identifying national health research priorities in Timor-Leste through a scoping review of existing health data.
Health research is crucial to understand a country's needs and to improve health outcomes. We conducted a scoping review and analysis of existing health data in Timor-Leste to identify the health research priorities of the country. Published and unpublished health research in Timor-Leste from 2001 to 2011 that reported objectives, methods and results were identified. Key findings were triangulated with data from national surveys and the Health Management Information System; 114 eligible articles were included in the analysis, the leading topics of which were communicable (malaria, tuberculosis, HIV and sexually transmitted diseases and dengue) and non-communicable (eye and mental health) diseases. There were 28 papers (25%) on safe motherhood, child health and nutrition, of which 20 (71%) were unpublished. The review of national indicators showed high infant, under-five and maternal mortality rates. Burden of disease is greatest in young children, with respiratory infections, febrile illnesses and diarrheal disease predominating. There is poor access to and utilization of health care. Childhood malnutrition is an important unresolved national health issue. There are several obstacles leading to under-utilization of health services. The following topics for future health research are suggested from the review: nutrition, safe motherhood, childhood illness (in particular identifying the causes and cause-specific burden of severe respiratory, febrile and diarrheal diseases) and access to and use of health services.


The effect of mutations in the lid region of Thermomyces lanuginosus lipase on interactions with triglyceride surfaces: A multi-scale simulation study.
Lipases naturally function at the interface formed between amphiphilic molecules and the aqueous environment. Thermomyces lanuginosus lipase (TLL) is a well-characterised lipase, known to exhibit interfacial activation during which a lid region covering the active site becomes displaced upon interaction with an interface. In this study, we investigate the effect the amino acid sequence of the lid region on interfacial binding and lid dynamics of TLL. Three TLL variants were investigated, a wild-type variant, a variant containing an esterase lid region (Esterase), and a Hybrid variant, containing both wild-type lid residues and esterase lid residues. Multiple coarse-grained molecular dynamics simulations revealed that the interfacial binding orientation of TLL was significantly affected by the nature of amino acids in the lid region, and atomistic simulations indicated effects on the structural dynamics of the lid itself. The atomistic simulations, as well as steered molecular dynamics simulations, also indicated that the Esterase lid region was less flexible than the wild-type lid region, whereas the Hybrid variant displayed superior lid flexibility and stability in the open conformation both at the interface, and in aqueous solution. Additional experiments performed to investigate the activity and binding behaviour of the lipase variants indicated a slightly higher specific activity for the Hybrid variant compared to the wild-type variant, correlating the observations of increased lid flexibility. Together, these results are in line with previous experimental studies, highlighting the importance of the nature of the amino acid residues within the functional lid region of lipases, particularly regarding interfacial binding orientation, activation, and structural stability.


Risk of cervical cancer among female autoworkers exposed to metalworking fluids.
Cervical cancer is caused by human papilloma virus (HPV). However, only a small proportion of women infected with HPV, progress to cervical cancer. Other co-factors must therefore be necessary to cause cervical cancer. We examined cervical cancer in relation to occupational exposure to metalworking fluids (MWF), which are complex mixtures containing several known carcinogens. A cohort of 4374 female autoworkers was followed from 1985-2004 for cancer diagnosis. Based on standardized incidence rates (SIR), we focused on cervical cancer in an internal analysis. Pooled logistic regression was used to model the relationship between exposure to three different types of MWF, selected constituents, and incidence of cervical cancer. Based on 40 cases, SIR were statistically significantly elevated for both race specific subgroups: 3.30 and 2.43, respectively for Caucasian and black women. The standard mortality ratio (SMR) was also statistically significantly elevated for Caucasian women (3.44) based on seven observed deaths. There was no association with oil-based straight fluid. Relative risks for soluble and synthetic MWF and nitrosamines were modestly elevated but not statistically significant. Water-based MWF may play a role in the etiology of cervical cancer. Further studies in larger cohorts of women are needed to clearly establish this relationship.


Measurement and modelling of deep sea sediment plumes and implications for deep sea mining.
Deep sea mining concerns the extraction of poly-metallic nodules, cobalt-rich crusts and sulphide deposits from the ocean floor. The exploitation of these resources will result in adverse ecological effects arising from the direct removal of the substrate and, potentially, from the formation of sediment plumes that could result in deposition of fine sediment on sensitive species or entrainment of sediment, chemicals and nutrients into over-lying waters. Hence, identifying the behaviour of deep-sea sediment plumes is important in designing mining operations that are ecologically acceptable. Here, we present the results of novel in situ deep sea plume experiments undertaken on the Tropic seamount, 300 nautical miles SSW of the Canary Islands. These plume experiments were accompanied by hydrographic and oceanographic field surveys and supported by detailed numerical modelling and high resolution video settling velocity measurements of the in situ sediment undertaken in the laboratory. The plume experiments involved the controlled formation of benthic sediment plumes and measurement of the plume sediment concentration at a specially designed lander placed at set distances from the plume origin. The experiments were used as the basis for validation of a numerical dispersion model, which was then used to predict the dispersion of plumes generated by full-scale mining. The results highlight that the extent of dispersion of benthic sediment plumes, resulting from mining operations, is significantly reduced by the effects of flocculation, background turbidity and internal tides. These considerations must be taken into account when evaluating the impact and extent of benthic sediment plumes.


5-Hydroxytryptamine type 2A receptors regulate cyclic AMP accumulation in a neuronal cell line by protein kinase C-dependent and calcium/calmodulin-dependent mechanisms.
The effects of 5-hydroxytryptamine (5-HT)2A receptor activation on cAMP formation were studied in a cell line derived from embryonic rat cortex (A1A1). 5-HT (EC50 = 0.87 microM) amplified the amount of cAMP formed in response to 5'-N-ethylcarboxamidoadenosine (an adenosine A2 receptor agonist), cholera toxin, and forskolin after 15 min of coincubation in the presence of the phosphodiesterase inhibitor rolipram. This effect of 5-HT was blocked by 10 nM ketanserin as well as by 10 nM spiperone, indicating a response mediated by the 5-HT2A receptor subtype. Similarly, cAMP accumulation was enhanced by coincubation with the protein kinase C (PKC) activator phorbol 12-myristate 13-acetate (PMA) and the calcium ionophore A23187. After exposure to PMA for 24 hr (PKC-depleted cells), 5-HT and A23187 still enhanced cAMP formed in response to forskolin and 5'-N-ethylcarboxamidoadenosine, whereas the amplifying effects of PMA were abolished. Analysis by Western blots and PKC activity measurements revealed that, of three PKC isoforms detected in A1A1 cells (alpha, delta, and epsilon), only the calcium-independent isoform PKC-epsilon remained in membrane fractions after long term PMA treatment. In PKC-depleted cells, 5-HT-mediated amplification was greatly reduced after treatment with the calcium chelator 1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (acetoxymethyl)-ester or the calmodulin antagonists calmidazolium and N-(6-aminohexyl)-5-chloro-1-napthalenesulfonamide hydrochloride. In addition, 5-HT-mediated amplification of cAMP accumulation was reduced by the PKC inhibitor staurosporine in normal cells but was unaffected in PKC-depleted cells. In conclusion, these data suggest that 5-HT2A receptor activation can amplify cAMP formation in A1A1 cells by two distinct pathways coupled to the hydrolysis of inositol phosphates, i.e., PKC and calcium/calmodulin.


Uncovering the dispersion history, adaptive evolution and selection of wheat in China.
Wheat was introduced to China approximately 4500 years ago, where it adapted over a span of time to various environments in agro-ecological growing zones. We investigated 717 Chinese and 14 Iranian/Turkish geographically diverse, locally adapted wheat landraces with 27 933 DArTseq (for 717 landraces) and 312 831 Wheat660K (for a subset of 285 landraces) markers. This study highlights the adaptive evolutionary history of wheat cultivation in China. Environmental stresses and independent selection efforts have resulted in considerable genome-wide divergence at the population level in Chinese wheat landraces. In total, 148 regions of the wheat genome show signs of selection in at least one geographic area. Our data show adaptive events across geographic areas, from the xeric northwest to the mesic south, along and among homoeologous chromosomes, with fewer variations in the D genome than in the A and B genomes. Multiple variations in interdependent functional genes such as regulatory and metabolic genes controlling germination and flowering time were characterized, showing clear allelic frequency changes corresponding to the dispersion of wheat in China. Population structure and selection data reveal that Chinese wheat spread from the northwestern Caspian Sea region to South China, adapting during its agricultural trajectory to increasingly mesic and warm climatic areas.


The role of fibronectin, laminin, vitronectin and their receptors on cellular adhesion in proliferative vitreoretinopathy.
To examine the possible role of some adhesion multifunctional glycoproteins of the extracellular matrix, such as fibronectin (FN), laminin (LN), vitronectin (VN) and their receptors (beta 1-subunit complex and alpha v beta 3 integrins) in events of cell migration and adhesion in proliferative vitreoretinopathy (PVR). Optical and electron-immunocytochemical techniques were carried out on epiretinal membranes. Electrophoretic immunoblotting methods and densitometric analysis of normal and PVR vitreous were also undertaken. Chi-square (chi 2) and unbalanced analysis of variance were employed for statistical analysis. FN was detected as a major component in the extracellular matrix in both fibrillar and pericellular arrangement. A change in pericellular distribution to more fibrillous organization was related to the time of intraocular proliferative tissue development (P < 0.001). LN and VN were observed as minor components in extracellular matrix. A colocalized pattern between VN and FN in collagenic bundles of the matrix was often observed. Beta-1 subunit and alpha v beta 3 receptors were usually localized in a position that could mediate the interaction of FN, VN, and/or LN to the cell plasma membrane. Increased levels of FN concentration were observed in both subretinal fluid and pathologic vitreous; intravitreal FN concentration tends to increase with clinical stages of the evolution of PVR, whereas intravitreal VN levels tend to decrease. Results suggest that FN could mediate the initial events involved in epiretinal membrane formation, and VN could modulate the adhesion mechanisms in established membranes.


Transcription profiling of peripheral B cells in antibody-positive primary Sjögren's syndrome reveals upregulated expression of CX3CR1 and a type I and type II interferon signature.
B cells play a key role in the pathogenesis of primary Sjögren's syndrome (pSS). The aim of this study was to analyse the transcriptome of CD19+ B cells from patients with pSS and healthy controls to decipher the B cell-specific contribution to pSS. RNA from purified CD19+ B cells from 12 anti-SSA antibody-positive untreated female patients with pSS and 20 healthy blood donors was subjected to whole transcriptome sequencing. A false discovery rate corrected significance threshold of α < 0.05 was applied to define differential gene expression. As validation, gene expression in B cells from 17 patients with pSS and 16 healthy controls was analysed using a targeted gene panel. RNA-sequencing identified 4047 differentially expressed autosomal genes in pSS B cells. Upregulated expression of type I and type II interferon (IFN)-induced genes was observed, establishing an IFN signature in pSS B cells. Among the top upregulated and validated genes were CX3CR1, encoding the fractalkine receptor involved in regulation of B-cell malignancies, CCL5/RANTES and CCR1. Increased expression of several members of the TNF superfamily was also identified; TNFSF4/Ox40L, TNFSF10/TRAIL, TNFSF13B/BAFF, TNFRSF17/BCMA as well as S100A8 and -A9/calprotectin, TLR7, STAT1 and STAT2. Among genes with downregulated expression in pSS B cells were SOCS1 and SOCS3, CD70 and TNFAIP3/A20. We conclude that B cells from patients with anti-SSA antibody-positive pSS display immune activation with upregulated expression of chemokines, chemokine receptors and a prominent type I and type II IFN signature, while suppressors of cytokine signalling are downregulated. This adds insight into the autoimmune process and suggests potential targets for future functional studies.


The tides of Titan.
We have detected in Cassini spacecraft data the signature of the periodic tidal stresses within Titan, driven by the eccentricity (e = 0.028) of its 16-day orbit around Saturn. Precise measurements of the acceleration of Cassini during six close flybys between 2006 and 2011 have revealed that Titan responds to the variable tidal field exerted by Saturn with periodic changes of its quadrupole gravity, at about 4% of the static value. Two independent determinations of the corresponding degree-2 Love number yield k(2) = 0.589 ± 0.150 and k(2) = 0.637 ± 0.224 (2σ). Such a large response to the tidal field requires that Titan's interior be deformable over time scales of the orbital period, in a way that is consistent with a global ocean at depth.


Prevalence of selected tick-borne pathogens in wild ungulates and ticks in southern Spain.
A survey study was carried out to assess the occurrence of selected tick-borne pathogens (TBP) in wild ungulates in Mediterranean ecosystems in southern Spain. Spleen samples were collected from 1,132 wild ungulates, including 578 red deer, 269 wild boar, 135 mouflon, 121 fallow deer and 29 roe deer, between 2009 and 2015. Eighty-nine ticks collected from TBP-positive animals were also analysed. Samples were tested by PCR and sequenced whenever possible. TBP DNA was detected in 127 of 863 wild ruminants (14.7%; 95% CI: 12.4-17.3) including the following: Anaplasma phagocytophilum (9.2%), Babesia divergens (2.9%), Theileria sp. OT3 (1.7%), Borrelia afzelii (0.7%) and Theileria capreoli (0.2%), but no positive samples were detected in wild boar (0/269). All the strains from mouflon were identified as Theileria sp. OT3, while B. divergens and T. capreoli were mainly found in red deer. Co-infection with A. phagocytophilum and B. divergens, and A. phagocytophilum and Theileria spp. was detected in red deer and mouflon, respectively. The risk factor analysis showed that the prevalences of A. phagocytophilum and piroplasms were species-related. Eighty-nine tick specimens collected from ungulates found to be infected with the selected TBP were identified as Hyalomma lusitanicum (95.5%) and Ixodes ricinus (4.5%). Thirty ticks were positive for Anaplasma/Ehrlichia spp. (33.7%), 25 for Babesia/Theileria (28.1%) and two for B. burgdorferi s.l. (2.3%). Eleven specimens showed co-infections with Anaplasma/Ehrlichia and Babesia/Theileria (10.1%) or Anaplasma/Ehrlichia and B. burgdorferi s.l. (2.3%). The estimated prevalences obtained in the present study suggest the possible contribution of wild ruminants to the maintenance of some selected TBP in Mediterranean ecosystems in southern Spain, while the role of wild boar in the epidemiology of these pathogens seems to be limited in this region.


Adipocyte-secreted exosomal microRNA-34a inhibits M2 macrophage polarization to promote obesity-induced adipose inflammation.
Persistent, unresolved inflammation in adipose tissue is a major contributor to obesity-associated metabolic complications. However, the molecular links between lipid-overloaded adipocytes and inflammatory immune cells in obese adipose tissues remain elusive. Here we identified adipocyte-secreted microRNA-34a (miR-34a) as a key mediator through its paracrine actions on adipose-resident macrophages. The expression of miR-34a in adipose tissues was progressively increased with the development of dietary obesity. Adipose-selective or adipocyte-specific miR-34a-KO mice were resistant to obesity-induced glucose intolerance, insulin resistance, and systemic inflammation, and this was accompanied by a significant shift in polarization of adipose-resident macrophages from proinflammatory M1 to antiinflammatory M2 phenotype. Mechanistically, mature adipocyte-secreted exosomes transported miR-34a into macrophages, thereby suppressing M2 polarization by repressing the expression of Krüppel-like factor 4 (Klf4). The suppressive effects of miR-34a on M2 polarization and its stimulation of inflammatory responses were reversed by ectopic expression of Klf4 in both bone marrow-derived macrophages and adipose depots of obese mice. Furthermore, increased miR-34a expression in visceral fat of overweight/obese subjects correlated negatively with reduced Klf4 expression, but positively with the parameters of insulin resistance and metabolic inflammation. In summary, miR-34a was a key component of adipocyte-secreted exosomal vesicles that transmitted the signal of nutrient overload to the adipose-resident macrophages for exacerbation of obesity-induced systemic inflammation and metabolic dysregulation.


Microcrustacean assemblages composition and environmental variables in lakes and ponds of the Andean region-South of Chile (37-39° S).
The zooplankton assemblages in Chilean Patagonian lakes are characterised mainly by their low biodiversity and high predominance of calanoids copepods, a pattern that has been studied for large and deep lakes between 38-51° S, and shallow ponds at 51° S. The aim of the present study was analyse the zooplankton assemblages in different water bodies located in coastal zones, middle valleys and mountain zones between 37-39° S. For this purpose, the following variables were considered: maximum depth, latitude, altitude, chlorophyll-a and species number, and to these variables, a Principal Component Analysis (PCA) was applied. A co-occurrence null model analysis was also applied for determining the existence of a random process in crustacean species associations. The results denoted low species richness, and different species associations for studied sites, and the null model analysis revealed the absence of a random process as regulator of species associations. Furthermore, a low species/genera ratio was identified, which denotes low productivity of the studied sites. These results were supported by a PCA analysis which denoted that the main determinant factors are chlorophyll concentration and species number that are directly associated. The obtained results are in agreement with descriptions in the literature for species diversity for lakes of Chilean Patagonia that describes oligotrophy as the main regulator of zooplankton assemblages. Other ecological and limnological topics are discussed in the present study.


Detection of small pancreatic tumors with multiphasic helical CT.
The purpose of this study was to evaluate the sensitivity and specificity of helical CT in the detection of adenocarcinomas of the pancreas measuring 2 cm or smaller at pathologic examination. Thin-section triple phase (20, 40, and 70 sec after the start of injection) contrast-enhanced helical CT scans of the abdomen in 18 patients with a pancreatic carcinoma that was 2 cm or smaller and 18 patients with a normal pancreas were retrospectively reviewed by two senior radiologists who specialized in oncologic abdominal imaging. Discrepancies were resolved by consensus. The observers were unaware of the clinical information. CT scans were evaluated for the presence of a pancreatic mass, bile, and pancreatic duct stricture. The location and size of tumors as determined on CT were compared with pathologic findings. The CT results were also compared with the prospective CT interpretations derived from the radiology reports and with the endoscopic sonographic reports when available. The sensitivity of thin-section triple-phase helical CT in the detection of small pancreatic masses was 77%, and the specificity was 100% for the two experienced observers. The sensitivity and specificity were 72% and 100%, respectively, for the prospective interpretations done by 10 observers. There was no correlation between the tumor size at pathology and the CT measurements. Thin-section contrast-enhanced helical CT is sensitive and highly specific for the detection of pancreatic tumors measuring 2 cm or smaller. Improvement in the detection rate of this technique compared with previous techniques lies in the optimization of parenchymal enhancement during the pancreatic phase and the decrease in slice thickness.


Motivational interviewing as a way to promote physical activity in obese adolescents: a randomised-controlled trial using self-determination theory as an explanatory framework.
Using self-determination theory (SDT) as an explanatory framework, this randomised-controlled study evaluates the effect of a motivational interviewing (MI)-based intervention as an addition to a standard weight loss programme (SWLP) on physical activity (PA) practice in obese adolescents over a six-month period. Fifty-four obese adolescents (mean age = 13 years, mean BMI = 29.57 kg/m²) were randomly assigned to an SWLP group (n = 28) or SWLP + MI group (n = 26). Both groups received two SWLP sessions, supplemented for the SWLP + MI group, by six MI sessions. Perceived autonomy support, perceived competence, motivational regulations, PA and BMI were assessed at baseline, three and six months (i.e. the end of the programme). MLM analyses revealed that compared to SWLP, the SWLP + MI group had a greater BMI decrease and a greater PA practice increase over time. Moreover, the SWLP + MI group reported greater autonomy support from medical staff at the end of the programme, greater increase in integrated and identified regulations and a stronger decrease in amotivation. MI appears as an efficient counselling method as an addition to an SWLP to promote PA in the context of pediatric obesity.


Insulin-induced vascular redox dysregulation in human atherosclerosis is ameliorated by dipeptidyl peptidase 4 inhibition.
Recent clinical trials have revealed that aggressive insulin treatment has a neutral effect on cardiovascular risk in patients with diabetes despite improved glycemic control, which may suggest confounding direct effects of insulin on the human vasculature. We studied 580 patients with coronary atherosclerosis undergoing coronary artery bypass surgery (CABG), finding that high endogenous insulin was associated with reduced nitric oxide (NO) bioavailability ex vivo in vessels obtained during surgery. Ex vivo experiments with human internal mammary arteries and saphenous veins obtained from 94 patients undergoing CABG revealed that both long-acting insulin analogs and human insulin triggered abnormal responses of post-insulin receptor substrate 1 downstream signaling ex vivo, independently of systemic insulin resistance status. These abnormal responses led to reduced NO bioavailability, activation of NADPH oxidases, and uncoupling of endothelial NO synthase. Treatment with an oral dipeptidyl peptidase 4 inhibitor (DPP4i) in vivo or DPP4i administered to vessels ex vivo restored physiological insulin signaling, reversed vascular insulin responses, reduced vascular oxidative stress, and improved endothelial function in humans. The detrimental effects of insulin on vascular redox state and endothelial function as well as the insulin-sensitizing effect of DPP4i were also validated in high-fat diet-fed ApoE-/- mice treated with DPP4i. High plasma DPP4 activity and high insulin were additively related with higher cardiac mortality in patients with coronary atherosclerosis undergoing CABG. These findings may explain the inability of aggressive insulin treatment to improve cardiovascular outcomes, raising the question whether vascular insulin sensitization with DPP4i should precede initiation of insulin treatment and continue as part of a long-term combination therapy.


Vagus nerve stimulation activates nucleus of solitary tract neurons via supramedullary pathways.
Vagus nerve stimulation (VNS) treats patients with drug-resistant epilepsy, depression and heart failure, but the mechanisms responsible are uncertain. The mild stimulus intensities used in chronic VNS suggest activation of myelinated primary visceral afferents projecting to the nucleus of the solitary tract (NTS). Here, we monitored the activity of second and higher order NTS neurons in response to peripheral vagal activation using therapeutic VNS criteria. A bipolar stimulating electrode activated the left cervical vagus nerve, and stereotaxically placed single tungsten electrodes recorded unit activity from the left caudomedial NTS of chloralose-anaesthetized rats. High-intensity single electrical stimuli established vagal afferent conduction velocity (myelinated A-type or unmyelinated C-type) as well as synaptic order (second vs. higher order using paired electrical stimuli) for inputs to single NTS neurons. Then, VNS treatment was applied. A mid-collicular knife cut (KC) divided the brainstem from all supramedullary regions to determine their contribution to NTS activity. Our chief findings indicate that the KC reduced basal spontaneous activity of second-order NTS neurons receiving myelinated vagal input by 85%. In these neurons, acute VNS increased activity similarly in Control and KC animals. Interestingly, the KC interrupted VNS activation of higher order NTS neurons and second-order NTS neurons receiving unmyelinated vagal input, indicating that supramedullary descending projections to NTS are needed to amplify the peripheral neuronal signal from VNS. The present study begins to define the pathways activated during VNS and will help to better identify the central nervous system contributions to the therapeutic benefits of VNS therapy. KEY POINTS: Vagus nerve stimulation is routinely used in the clinic to treat epilepsy and depression, despite our uncertainty about how this treatment works. For this study, the connections between the nucleus of the solitary tract (NTS) and the higher brain regions were severed to learn more about their contribution to activity of these neurons during stimulation. Severing these brain connections reduced baseline activity as well as reducing stimulation-induced activation for NTS neurons receiving myelinated vagal input. Higher brain regions play a significant role in maintaining both normal activity in NTS and indirect mechanisms of enhancing NTS neuronal activity during vagus nerve stimulation.


Sustained CD8+ T cell memory inflation after infection with a single-cycle cytomegalovirus.
Cytomegalovirus (CMV) is a β-herpesvirus that establishes a lifelong latent or persistent infection. A hallmark of chronic CMV infection is the lifelong persistence of large numbers of virus-specific CD8+ effector/effector memory T cells, a phenomenon called "memory inflation". How the virus continuously stimulates these T cells without being eradicated remains an enigma. The prevailing view is that CMV establishes a low grade "smoldering" infection characterized by tiny bursts of productive infection which are rapidly extinguished, leaving no detectable virus but replenishing the latent pool and leaving the immune system in a highly charged state. However, since abortive reactivation with limited viral gene expression is known to occur commonly, we investigated the necessity for virus reproduction in maintaining the inflationary T cell pool. We inhibited viral replication or spread in vivo using two different mutants of murine CMV (MCMV). First, famcyclovir blocked the replication of MCMV encoding the HSV Thymidine Kinase gene, but had no impact on the CD8+ T cell memory inflation once the infection was established. Second, MCMV that lacks the essential glycoprotein L, and thus is completely unable to spread from cell to cell, also drove memory inflation if the virus was administered systemically. Our data suggest that CMV which cannot spread from the cells it initially infects can repeatedly generate viral antigens to drive memory inflation without suffering eradication of the latent genome pool.


Multiple gene genealogical analyses reveal both common and distinct population genetic patterns among replicons in the nitrogen-fixing bacterium Sinorhizobium meliloti.
Sinorhizobium meliloti is a Gram-negative alpha-proteobacterium that can form symbiotic relationships with alfalfa and fix atmospheric nitrogen. The complete genome of a laboratory strain, Rm1021, was published in 2001, and the genome of this strain is arranged in three replicons: a chromosome of 3.65 million base pairs (Mb), and two megaplasmids, pSymA (1.35 Mb) and pSymB (1.68 Mb). However, the potential difference in genetic variation among the three replicons in natural strains remains poorly understood. In this study, a total of 16 gene fragments were sequenced, four from pSymA and six each from the chromosome and pSymB, for 49 natural S. meliloti strains. The analyses identified significant differences in divergence among genes, with the mean Hasegawa-Kishino-Yano-1985 (HKY85) distance ranging from 0.00157 to 0.04109 between pairs of strains. Overall, genes on pSymA showed the highest mean HKY85 distance, followed by those on pSymB and the chromosome. Although evidence for recombination was found, the authors' population genetic analyses revealed overall significant linkage disequilibria among genes within both pSymA and the chromosome. However, genes on pSymB were in overall linkage equilibrium, consistent with frequent recombination among genes on this replicon. Furthermore, the genealogical comparisons among the three replicons identified significant incongruence, indicating reassortment among the three replicons in natural populations. The results suggest both shared and distinct patterns of molecular evolution among the three replicons in the genomes of natural strains of S. meliloti.


Standardized computer-based organized reporting of EEG: SCORE.
The electroencephalography (EEG) signal has a high complexity, and the process of extracting clinically relevant features is achieved by visual analysis of the recordings. The interobserver agreement in EEG interpretation is only moderate. This is partly due to the method of reporting the findings in free-text format. The purpose of our endeavor was to create a computer-based system for EEG assessment and reporting, where the physicians would construct the reports by choosing from predefined elements for each relevant EEG feature, as well as the clinical phenomena (for video-EEG recordings). A working group of EEG experts took part in consensus workshops in Dianalund, Denmark, in 2010 and 2011. The faculty was approved by the Commission on European Affairs of the International League Against Epilepsy (ILAE). The working group produced a consensus proposal that went through a pan-European review process, organized by the European Chapter of the International Federation of Clinical Neurophysiology. The Standardised Computer-based Organised Reporting of EEG (SCORE) software was constructed based on the terms and features of the consensus statement and it was tested in the clinical practice. The main elements of SCORE are the following: personal data of the patient, referral data, recording conditions, modulators, background activity, drowsiness and sleep, interictal findings, "episodes" (clinical or subclinical events), physiologic patterns, patterns of uncertain significance, artifacts, polygraphic channels, and diagnostic significance. The following specific aspects of the neonatal EEGs are scored: alertness, temporal organization, and spatial organization. For each EEG finding, relevant features are scored using predefined terms. Definitions are provided for all EEG terms and features. SCORE can potentially improve the quality of EEG assessment and reporting; it will help incorporate the results of computer-assisted analysis into the report, it will make possible the build-up of a multinational database, and it will help in training young neurophysiologists.


BAY 43-9006 exhibits broad spectrum oral antitumor activity and targets the RAF/MEK/ERK pathway and receptor tyrosine kinases involved in tumor progression and angiogenesis.
The RAS/RAF signaling pathway is an important mediator of tumor cell proliferation and angiogenesis. The novel bi-aryl urea BAY 43-9006 is a potent inhibitor of Raf-1, a member of the RAF/MEK/ERK signaling pathway. Additional characterization showed that BAY 43-9006 suppresses both wild-type and V599E mutant BRAF activity in vitro. In addition, BAY 43-9006 demonstrated significant activity against several receptor tyrosine kinases involved in neovascularization and tumor progression, including vascular endothelial growth factor receptor (VEGFR)-2, VEGFR-3, platelet-derived growth factor receptor beta, Flt-3, and c-KIT. In cellular mechanistic assays, BAY 43-9006 demonstrated inhibition of the mitogen-activated protein kinase pathway in colon, pancreatic, and breast tumor cell lines expressing mutant KRAS or wild-type or mutant BRAF, whereas non-small-cell lung cancer cell lines expressing mutant KRAS were insensitive to inhibition of the mitogen-activated protein kinase pathway by BAY 43-9006. Potent inhibition of VEGFR-2, platelet-derived growth factor receptor beta, and VEGFR-3 cellular receptor autophosphorylation was also observed for BAY 43-9006. Once daily oral dosing of BAY 43-9006 demonstrated broad-spectrum antitumor activity in colon, breast, and non-small-cell lung cancer xenograft models. Immunohistochemistry demonstrated a close association between inhibition of tumor growth and inhibition of the extracellular signal-regulated kinases (ERKs) 1/2 phosphorylation in two of three xenograft models examined, consistent with inhibition of the RAF/MEK/ERK pathway in some but not all models. Additional analyses of microvessel density and microvessel area in the same tumor sections using antimurine CD31 antibodies demonstrated significant inhibition of neovascularization in all three of the xenograft models. These data demonstrate that BAY 43-9006 is a novel dual action RAF kinase and VEGFR inhibitor that targets tumor cell proliferation and tumor angiogenesis.


Reduced graphene oxide nanosheets coated with an anti-angiogenic anticancer low-molecular-weight heparin derivative for delivery of anticancer drugs.
Here, we report reduced graphene oxide (rGO) nanosheets coated with an anti-angiogenic anticancer taurocholate derivative of low-molecular-weight heparin (LHT7) as a tumor-targeting nanodelivery platform for anticancer drugs. Surface coating of LHT7 onto rGO was confirmed using fluorescein isothiocyanate-labeled LHT7, monitored as fluorescence quenching due to associated rGO. Unlike plain rGO, LHT7-coated rGO (LHT-rGO) nanosheets maintained a stable dispersion under physiological conditions for at least 24h. Moreover, LHT-rGO provided greater loading capacity for doxorubicin (Dox) compared with uncoated rGO nanosheets. Following intravenous administration into KB tumor-bearing mice, in vivo tumor accumulation of LHT-rGO/Dox was 7-fold higher than that of rGO/Dox 24h post dosing. In tumor tissues, LHT-rGO/Dox was shown to localize not to the tumor vasculature, but rather to tumor cells. Intravenously administered LHT-rGO/Dox showed the greatest anti-tumor effect in KB-bearing mice, reducing tumor volume by 92.5%±3.1% compared to the untreated group 25days after tumor inoculation. TUNEL assays revealed that the population of apoptotic cells was highest in the group treated with LHT-rGO/Dox. Taken together, our results demonstrate that LHT-rGO nanosheets confer improved dispersion stability, tumor distribution and in vivo antitumor effects, and may be further developed as a potential active nanoplatform of various anticancer drugs.


Epidemiology and clinical associations of human parechovirus respiratory infections.
Infections with human parechoviruses (HPeVs) are prevalent in young children and have been associated with mild gastroenteritis and, less frequently, with meningitis and neonatal sepsis. To investigate the involvement of these viruses in respiratory disease, a highly sensitive nested PCR was used to screen a large archive of respiratory specimens, collected between January and December 2007. Respiratory samples had previously been tested for eight respiratory viruses, including respiratory syncytial virus and adenovirus, by PCR. HPeV was detected in 34 of 3,844 specimens, representing 27 of 2,220 study subjects (1.2%). HPeV types were identified by sequencing the VP3/VP1 junction amplified by PCR directly from clinical specimens. The assay could amplify all HPeV types examined with high sensitivity (types 1 and 3 to 6) and also identified HPeV types in all but one of the screen-positive study specimens (25 HPeV1 and eight HPeV6 specimens). Infections with both HPeV1 and HPeV6 were seasonal, with highest frequencies in July and August, and restricted to children aged between 6 months and 5 years. Other respiratory viruses were frequently codetected in HPeV-positive specimens, with significant overrepresentation of adenovirus coinfections (37%). Most HPeV-positive specimens were referred from emergency departments, although no association with specific respiratory symptoms or disease was found. In summary, the low frequency of detection and lack of clear disease associations indicate that HPeV1 and -6 are not major pathogens in individuals presenting with respiratory disease. However, the screening and typing methods developed will be of value in further HPeV testing, including testing for meningitis cases and other suspected HPeV-associated disease presentations.


Neuronal KCNQ potassium channels: physiology and role in disease.
Humans have over 70 potassium channel genes, but only some of these have been linked to disease. In this respect, the KCNQ family of potassium channels is exceptional: mutations in four out of five KCNQ genes underlie diseases including cardiac arrhythmias, deafness and epilepsy. These disorders illustrate the different physiological functions of KCNQ channels, and provide a model for the study of the 'safety margin' that separates normal from pathological levels of channel expression. In addition, several KCNQ isoforms can associate to form heteromeric channels that underlie the M-current, an important regulator of neuronal excitability.


Buffer capacity of biologics--from buffer salts to buffering by antibodies.
Controlling pH is essential for a variety of biopharmaceutical process steps. The chemical stability of biologics such as monoclonal antibodies is pH-dependent and slightly acidic conditions are favorable for stability in a number of cases. Since control of pH is widely provided by added buffer salts, the current study summarizes the buffer characteristics of acetate, citrate, histidine, succinate, and phosphate buffers. Experimentally derived values largely coincide with values calculated from a model that had been proposed in 1922 by van Slyke. As high concentrated protein formulations become more and more prevalent for biologics, the self-buffering potential of proteins becomes of relevance. The current study provides information on buffer characteristics for pH ranges down to 4.0 and up to 8.0 and shows that a monoclonal antibody at 50 mg/mL exhibits similar buffer capacity as 6 mM citrate or 14 mM histidine (pH 5.0-6.0). Buffer capacity of antibody solutions scales linearly with protein concentration up to more than 200 mg/mL. At a protein concentration of 220 mg/mL, the buffer capacity resembles the buffer capacity of 30 mM citrate or 50 mM histidine (pH 5.0-6.0). The buffer capacity of monoclonal antibodies is practically identical at the process relevant temperatures 5, 25, and 40°C. Changes in ionic strength of ΔI=0.15, in contrast, can alter the buffer capacity up to 35%. In conclusion, due to efficient self-buffering by antibodies in the pH range of favored chemical stability, conventional buffer excipients could be dispensable for pH stabilization of high concentrated protein solutions.


[Not Available].
The survey "Datenerhebung zur Depression in der Allgemeinbevölkerung" was conducted from fall 2010 to spring 2011 on several online platforms. The results show that there is a considerable timespan between the appearance of initial symptoms of depression and the first diagnosis of a patient. Intervention at early stages of the disease can reduce a potentially long time of suffering and can lead to the successful treatment of depression. General practitioners play an important role as the link between patient and psychiatrist.


Genotoxic exposures of potroom workers.
Potroom workers in aluminum reduction plants have increased risks for bladder and lung cancer due to exposure from polycyclic aromatic hydrocarbons (PAH). In this study correlations between measures of the external, internal, and biological effective dose have been studied for PAH. Venous blood samples were obtained from 98 male potroom workers and 55 unexposed male blue-collar workers, for the analysis of aromatic adducts to DNA (deoxyribonucleic acid) in lymphocytes, using the 32P-postlabeling technique. 1-Hydroxypyrene in urine was analyzed with high-pressure liquid chromatography. Personal sampling of both particulate and gas phase PAH was performed during a full workday for the potroom workers and for 5 referents. Individual PAH congeners were determined with liquid chromatographic-mass spectrometric and gas chromatographic-mass spectrometric techniques. The respiratory-zone airborne level of the sum of 22 particulate (median 13.2 micro/m3) and the 7 gas phase PAH-congeners (median 16.3 microg/m3) among the potroom workers was a hundred times higher than among the referents. The urinary concentration of 1-hydroxypyrene before work was 30 times higher for the potroom workers (median 3.43 micromol/mol creatinine) than for the referents. Most airborne PAH congeners correlated with the excretion of 1-hydroxypyrene in urine. The frequency of aromatic DNA adducts did not, however, differ between the potroom workers and the referents, and no correlation was found for 1-hydroxypyrene in urine. Despite an obvious occupational exposure to PAH, no increase in aromatic DNA adducts in lymphocytes was found among the potroom workers.


In situ characterization of crystallization and melting of soft, thermoresponsive microgels by small-angle X-ray scattering.
Depending on the volume fraction and interparticle interactions, colloidal suspensions can form different phases, ranging from fluids, crystals, and glasses to gels. For soft microgels that are made from thermoresponsive polymers, the volume fraction can be tuned by temperature, making them excellent systems to experimentally study phase transitions in dense colloidal suspensions. However, investigations of phase transitions at high particle concentration and across the volume phase transition temperature in particular, are challenging due to the deformability and possibility for interpenetration between microgels. Here, we investigate the dense phases of composite core-shell microgels that have a small gold core and a thermoresponsive microgel shell. Employing Ultra Small-Angle X-ray Scattering, we make use of the strong scattering signal from the gold cores with respect to the almost negligible signal from the shells. By changing the temperature we study the freezing and melting transitions of the system in situ. Using Bragg peak analysis and the Williamson-Hall method, we characterize the phase transitions in detail. We show that the system crystallizes into an rhcp structure with different degrees of in-plane and out-of-plane stacking disorder that increase upon particle swelling. We further find that the melting process is distinctly different, where the system separates into two different crystal phases with different melting temperatures and interparticle interactions.


Local breast cancer spatial patterning: a tool for community health resource allocation to address local disparities in breast cancer mortality.
Despite available demographic data on the factors that contribute to breast cancer mortality in large population datasets, local patterns are often overlooked. Such local information could provide a valuable metric by which regional community health resources can be allocated to reduce breast cancer mortality. We used national and statewide datasets to assess geographical distribution of breast cancer mortality rates and known risk factors influencing breast cancer mortality in middle Tennessee. Each county in middle Tennessee, and each ZIP code within metropolitan Davidson County, was scored for risk factor prevalence and assigned quartile scores that were used as a metric to identify geographic areas of need. While breast cancer mortality often correlated with age and incidence, geographic areas were identified in which breast cancer mortality rates did not correlate with age and incidence, but correlated with additional risk factors, such as mammography screening and socioeconomic status. Geographical variability in specific risk factors was evident, demonstrating the utility of this approach to identify local areas of risk. This method revealed local patterns in breast cancer mortality that might otherwise be overlooked in a more broadly based analysis. Our data suggest that understanding the geographic distribution of breast cancer mortality, and the distribution of risk factors that contribute to breast cancer mortality, will not only identify communities with the greatest need of support, but will identify the types of resources that would provide the most benefit to reduce breast cancer mortality in the community.


Mitogenomics reveals high synteny and long evolutionary histories of sympatric cryptic nematode species.
Species with seemingly identical morphology but with distinct genetic differences are abundant in the marine environment and frequently co-occur in the same habitat. Such cryptic species are typically delineated using a limited number of mitochondrial and/or nuclear marker genes, which do not yield information on gene order and gene content of the genomes under consideration. We used next-generation sequencing to study the composition of the mitochondrial genomes of four sympatrically distributed cryptic species of the Litoditis marina species complex (PmI, PmII, PmIII, and PmIV). The ecology, biology, and natural occurrence of these four species are well known, but the evolutionary processes behind this cryptic speciation remain largely unknown. The gene order of the mitochondrial genomes of the four species was conserved, but differences in genome length, gene length, and codon usage were observed. The atp8 gene was lacking in all four species. Phylogenetic analyses confirm that PmI and PmIV are sister species and that PmIII diverged earliest. The most recent common ancestor of the four cryptic species was estimated to have diverged 16 MYA. Synonymous mutations outnumbered nonsynonymous changes in all protein-encoding genes, with the Complex IV genes (coxI-III) experiencing the strongest purifying selection. Our mitogenomic results show that morphologically similar species can have long evolutionary histories and that PmIII has several differences in genetic makeup compared to the three other species, which may explain why it is better adapted to higher temperatures than the other species.


Real-time computation at the edge of chaos in recurrent neural networks.
Depending on the connectivity, recurrent networks of simple computational units can show very different types of dynamics, ranging from totally ordered to chaotic. We analyze how the type of dynamics (ordered or chaotic) exhibited by randomly connected networks of threshold gates driven by a time-varying input signal depends on the parameters describing the distribution of the connectivity matrix. In particular, we calculate the critical boundary in parameter space where the transition from ordered to chaotic dynamics takes place. Employing a recently developed framework for analyzing real-time computations, we show that only near the critical boundary can such networks perform complex computations on time series. Hence, this result strongly supports conjectures that dynamical systems that are capable of doing complex computational tasks should operate near the edge of chaos, that is, the transition from ordered to chaotic dynamics.


Shaping Ability of Reciproc, WaveOne GOLD, and HyFlex EDM Single-file Systems in Simulated S-shaped Canals.
The aim of the present study was to compare the shaping ability of Reciproc (RPC; VDW, Munich, Germany), HyFlex EDM (HEDM; Coltene/Whaledent AG, Altstätten, Switzerland), and WaveOne GOLD (WOG; Dentsply Maillefer, Ballaigues, Switzerland) nickel-titanium (NiTi) files made of different NiTi alloys in S-shaped simulated canals. Sixty S-shaped canals in resin blocks were prepared to an apical size of 0.25 mm using RPC R25, WOG Primary, and HEDM OneFile (n = 20 canal/per group) systems. Composite images were made from the superimposition of pre- and postinstrumentation images. The amount of resin removed by each system was measured using a digital template and image analysis software in 22 different points. Canal aberrations were also recorded. Data were statistically analyzed using the Kruskal-Wallis and post hoc Dunn tests at the 5% level. NiTi file fracture was not observed during shaping of the simulated canals although a danger zone formation in 1 sample and a ledge in 1 sample were observed in the RPC group. There was no statistically significant difference between the WOG and HEDM groups' apical, medial, and coronal regions (P > .05). However, it was determined that the RPC group removed a statistically significantly higher amount of resin from all the canal regions when compared with the WOG and HEDM groups (P < .05). Within the limitation of the present study, it was determined that all of the tested NiTi files caused various levels of resin removal. However, WOG and HEDM NiTi files were found to cause a lower level of resin removal than RPC NiTi files.


Collisional quenching of erythrosine B as a potential reference dye for impulse response function evaluation.
We studied the collisional quenching of the erythrosine B fluorophore by potassium iodide. The quenching follows a Stern-Volmer dependence up to the highest quencher concentration. The lifetime of erythrosine B decreases to 24 ps in 5.02 M of potassium iodide. The quantum yield of erythrosine B in the presence of 5.02 M KI is 0.0035. The relatively high brightness makes this compound attractive as an ultrashort reference in time-resolved measurements. In both frequency- and time-domain fluorescence techniques, there is a need for lifetime standards with extremely short decay times. Mimicking the instantaneous scattering at longer wavelengths allows color-effect-free measurements in the emission region. Another motivation is the problem of obtaining the impulse response function in the case of two-photon excitation. Time-resolved microscopy also benefits from fast-decaying dyes because the impulse response function can be evaluated at the emission wavelength of the investigated specimen without changing filters. We demonstrated that impulse response functions for commonly used detectors are practically the same for scattering as for quenched erythrosine B emission. We also analyzed a complex fluorescence decay using both elastic scattering and quenched erythrosine B emission as a response function.


Kinetic parameters for nutrient enhanced crude oil biodegradation in intertidal marine sediments.
Availability of inorganic nutrients, particularly nitrogen and phosphorous, is often a primary control on crude oil hydrocarbon degradation in marine systems. Many studies have empirically determined optimum levels of inorganic N and P for stimulation of hydrocarbon degradation. Nevertheless, there is a paucity of information on fundamental kinetic parameters for nutrient enhanced crude oil biodegradation that can be used to model the fate of crude oil in bioremediation programmes that use inorganic nutrient addition to stimulate oil biodegradation. Here we report fundamental kinetic parameters (Ks and qmax) for nitrate- and phosphate-stimulated crude oil biodegradation under nutrient limited conditions and with respect to crude oil, under conditions where N and P are not limiting. In the marine sediments studied, crude oil degradation was limited by both N and P availability. In sediments treated with 12.5 mg/g of oil but with no addition of N and P, hydrocarbon degradation rates, assessed on the basis of CO2 production, were 1.10 ± 0.03 μmol CO2/g wet sediment/day which were comparable to rates of CO2 production in sediments to which no oil was added (1.05 ± 0.27 μmol CO2/g wet sediment/day). When inorganic nitrogen was added alone maximum rates of CO2 production measured were 4.25 ± 0.91 μmol CO2/g wet sediment/day. However, when the same levels of inorganic nitrogen were added in the presence of 0.5% P w/w of oil (1.6 μmol P/g wet sediment) maximum rates of measured CO2 production increased more than four-fold to 18.40 ± 1.04 μmol CO2/g wet sediment/day. Ks and qmax estimates for inorganic N (in the form of sodium nitrate) when P was not limiting were 1.99 ± 0.86 μmol/g wet sediment and 16.16 ± 1.28 μmol CO2/g wet sediment/day respectively. The corresponding values for P were 63 ± 95 nmol/g wet sediment and 12.05 ± 1.31 μmol CO2/g wet sediment/day. The qmax values with respect to N and P were not significantly different (P < 0.05). When N and P were not limiting Ks and qmax for crude oil were 4.52 ± 1.51 mg oil/g wet sediment and 16.89 ± 1.25 μmol CO2/g wet sediment/day. At concentrations of inorganic N above 45 μmol/g wet sediment inhibition of CO2 production from hydrocarbon degradation was evident. Analysis of bacterial 16S rRNA genes indicated that Alcanivorax spp. were selected in these marine sediments with increasing inorganic nutrient concentration, whereas Cycloclasticus spp. were more prevalent at lower inorganic nutrient concentrations. These data suggest that simple empirical estimates of the proportion of nutrients added relative to crude oil concentrations may not be sufficient to guarantee successful crude oil bioremediation in oxic beach sediments. The data we present also help define the maximum rates and hence timescales required for bioremediation of beach sediments.


Supercritical xenon-filled hollow-core photonic bandgap fiber.
We demonstrate that filling a hollow-core photonic-bandgap fiber with supercritical xenon creates a medium with a controllable density up to several hundred times that at STP, while working at room temperature. The high compressibility of the supercritical fluid allows rapid tuning of the spectral guidance window by making small changes of gas pressure near the critical point. We discuss potential applications of this system in linear and nonlinear optics.


Electro-optic response of second-harmonic generation membrane potential sensors.
We quantify and ascertain the nature of the second-harmonic generation (SHG) response of amphiphilic push-pull chromophores to a transmembrane electric field. Our technique is based on the application of an alternating field across labeled giant unilamelar vesicles. For chromophore responses that are purely electro-optic, our technique provides an estimate of photoinduced charge shifts based on the observed dispersion of the field response, in accord with a two-level perturbation theory. These results are applicable to the optimization of membrane potential sensors for SHG microscopy.


Strong association between glucocerebrosidase mutations and Parkinson's disease in Sweden.
Several genetic studies have demonstrated an association between mutations in glucocerebrosidase (GBA), originally implicated in Gaucher's disease, and an increased risk of Parkinson's disease (PD). We have investigated the possible involvement of genetic GBA variations in PD in the Swedish population. Three GBA variants, E326K, N370S, and L444P were screened in the largest Swedish Parkinson cohort reported to date; 1625 cases and 2025 control individuals. We found a significant association with high effect size of the rare variant L444P with PD (odds ratio 8.17; 95% confidence interval: 2.51-26.23; p-value = 0.0020) and a significant association of the common variant E326K (odds ratio 1.60; 95% confidence interval: 1.16-2.22; p-value = 0.026). The rare variant N370S showed a trend for association. Most L444P carriers (68%) were found to reside in northern Sweden, which is consistent with a higher prevalence of Gaucher's disease in this part of the country. Our findings support the role of GBA mutations as risk factors for PD and point to lysosomal dysfunction as a mechanism contributing to PD etiology.


Theoretical identification of seven C80 fullerene isomers by XPS and NEXAFS spectroscopy.
The carbon K-shell (1s) X-ray photoelectron spectroscopy (XPS) and near-edge X-ray absorption fine structure (NEXAFS) spectroscopy of the seven isolated-pentagon-rule (IPR) isomers of fullerene C80 have been calculated by means of density functional theory (DFT) theoretically. We have demonstrated the relationship between molecular structures and related X-ray spectroscopies. The dependence of the XPS spectra on the structures of the seven C80 molecules is imperfect, while the NEXAFS spectra show strong dependence on the seven fullerene molecules, so the NEXAFS spectra can be employed to identify all of the studied isomers. The spectral components of different local environments have been explored in detail.


Predictors of Complications of Tonsillectomy With or Without Adenoidectomy in Hospitalized Children and Adolescents in the United States, 2001-2010: A Population-Based Study.
Outcomes of tonsillectomy (with or without adenoidectomy [w/woA]) in hospitalized children are unclear. We sought, to describe the characteristics of hospitalized children who underwent tonsillectomy (w/woA), to estimate the prevalence of complications and to evaluate the relative impact of different comorbid conditions (CMC) on the risk of occurrence of common complications following these procedures. All patients aged ≤21years who underwent a tonsillectomy (w/woA) were selected from the Nationwide Inpatient Sample (NIS 2001-2010). The associations between several patient/hospital-level factors and occurrence of complications were generated using multivariable logistic regression models. Over a decade, a total of 141 599 hospitalized patients underwent tonsillectomy (w116 319; woA 25 280). A total of 58.1% were males. Majority of the procedures were performed in teaching hospitals (TH, 73.7%), in large (bed-size) hospitals (LH, 57.8%), and in those who were electively admitted (EA, 67.3%). Frequently present CMC in patients included obstructive sleep apnea (OSA, 26.4%), chronic pulmonary disease (CPD, 14.6%), neurological disorders (ND, 6.7%), and obesity (4.8%). Majority of patients were discharged routinely (98%). Overall complication rate was 6.4% with common complications being postoperative pneumonia (2.3%), bacterial infections (1.4%), respiratory complications (1.3%), and hemorrhage (1.2%). All-cause mortality included a total of 60 patients. Patients in TH (odds ratio [OR] = 0.72, 95%CI = 0.62-0.85), LH (OR = 0.80, 95% CI = 0.69-0.93), and those who had the procedures during EA (OR = 0.64, 95% CI = 0.56-0.74) had significantly lower odds of complications compared with their counterparts. CMC such as anemia, CPD, coagulopathy, HT, ND, and fluid/electrolyte disorders were independent predictors of significantly higher complication risk (P < .05). In conclusion, hospitalized children who underwent tonsillectomy (w/woA) in large or teaching hospitals, or during elective admissions had lower risk of complications. Comorbidity is an important independent predictor of complications in this cohort.


A function for phosphatidylinositol 3-kinase beta (p85alpha-p110beta) in fibroblasts during mitogenesis: requirement for insulin- and lysophosphatidic acid-mediated signal transduction.
We have previously shown that phosphatidylinositol 3-kinase alpha (PI 3-Kalpha) (p85alpha-p110alpha) is required for DNA synthesis induced by various growth factors (S. Roche, M. Koegl, and S. A. Courtneidge, Proc. Natl. Acad. Sci. USA 91:9185-9189, 1994) in fibroblasts. In the present study, we have investigated the function of PI 3-Kbeta (p85alpha-p110beta) during mitogenesis. By using antibodies specific to p110beta we showed that PI 3-Kbeta is expressed in NIH 3T3 cells. PI 3-Kbeta and PI 3-Kalpha have common features: PI 3-Kbeta is tightly associated with a protein serine kinase that phosphorylates p85alpha, it interacts with the Src-middle T antigen complex and the activated platelet-derived growth factor (PDGF) receptor in fibroblasts in vivo, and it becomes tyrosine phosphorylated after PDGF stimulation. PI 3-Kbeta was also activated in Swiss 3T3 and Cos7 cells stimulated with lysophosphatidic acid (LPA), a mitogen that interacts with a heterotrimeric G protein-coupled receptor. In contrast PI 3-Kalpha was activated to a lesser extent in these cells. Microinjection of neutralizing antibodies specific for p110beta into quiescent fibroblasts inhibited DNA synthesis induced by both insulin and LPA but poorly affected PDGF receptor signaling. Therefore, PI 3-Kbeta plays an important role in transmitting the mitogenic response induced by some, but not all, growth factors. Finally, we show that while oncogenic V12Ras interacts with type I PI 3-Ks, it could induce DNA synthesis in the absence of active PI 3-Kalpha and PI 3-Kbeta, suggesting that Ras uses other effectors for DNA synthesis.


Verapamil prevents slowing of transmural conduction and suppresses arrhythmias in an isolated guinea pig ventricular model of ischemia and reperfusion.
Transmembrane electrical activity was recorded from endocardium and epicardium of isolated segments of guinea pig right ventricular free walls. An electrocardiogram was recorded by electrodes at opposite ends of the tissue bath. Endocardium was stimulated. Tissues were exposed to "ischemic" conditions (e.g., acidosis, hyperkalemia, hypoxia, and lactate) for 15 minutes and then were reperfused with "normal" Tyrode's solution. Arrhythmias with characteristics of transmural reentry occurred in ischemic conditions and early reperfusion in 30% and 70% of 20 control hearts, respectively. Arrhythmias were associated with prolongation of transmural conduction time (CT) and abbreviation of endocardial effective refractory period. Verapamil significantly suppressed reperfusion arrhythmias at 0.1-1.0 microM but not at 3.0 microM. Verapamil also significantly decreased the incidence of arrhythmias during ischemic conditions at 0.5 microM but significantly promoted ischemic arrhythmias at 3.0 microM. Action potential duration and effective refractory period were not altered by verapamil during ischemic conditions or reperfusion. However, at 0.1-1.0 microM, verapamil prevented or attenuated prolongation of transmural CT by ischemic conditions and reperfusion. Transmural CT was further prolonged at 3 microM verapamil. In epicardial slices, 1 microM verapamil shortened CT transverse to fiber orientation during reperfusion but had no effect on longitudinal CT. Our results indicate that verapamil may suppress arrhythmias through differential effects on CT transverse and longitudinal to fiber orientation in anisotropic ventricular tissues and thus by specifically improving transmural conduction.


School-age children's environmental object identification in natural auditory scenes: effects of masking and contextual congruence.
This study investigated the development of children's skills in identifying ecologically relevant sound objects within naturalistic listening environments, using a non-linguistic analog of the classic 'cocktail-party' situation. Children aged 7-12.5 years completed a closed-set identification task in which brief, commonly encountered environmental sounds were presented at varying signal-to-noise ratios. To simulate the complexity of real-world acoustic environments, target sounds were embedded in either a single, stereophonically presented scene, or in one of two different scenes, with each scene presented to a single ear. Each target sound was either congruent or incongruent with the auditory context. Identification accuracy improved with increasing age, particularly in trials with low signal-to-noise ratios. Performance was most accurate when target sounds were incongruent with the background scene, and when sounds were presented in a single background scene. The presence of two backgrounds disproportionately disrupted children's performance relative to that of previously tested adults, and reduced children's sensitivity to contextual cues. Successful identification of familiar sounds in complex auditory contexts is the outcome of a protracted learning process, with children reaching adult levels of performance after a decade or more of experience.


Scientists' warning on extreme wildfire risks to water supply.
2020 is the year of wildfire records. California experienced its three largest fires early in its fire season. The Pantanal, the largest wetland on the planet, burned over 20% of its surface. More than 18 million hectares of forest and bushland burned during the 2019-2020 fire season in Australia, killing 33 people, destroying nearly 2500 homes, and endangering many endemic species. The direct cost of damages is being counted in dozens of billion dollars, but the indirect costs on water-related ecosystem services and benefits could be equally expensive, with impacts lasting for decades. In Australia, the extreme precipitation ("200 mm day -1 in several location") that interrupted the catastrophic wildfire season triggered a series of watershed effects from headwaters to areas downstream. The increased runoff and erosion from burned areas disrupted water supplies in several locations. These post-fire watershed hazards via source water contamination, flash floods, and mudslides can represent substantial, systemic long-term risks to drinking water production, aquatic life, and socio-economic activity. Scenarios similar to the recent event in Australia are now predicted to unfold in the Western USA. This is a new reality that societies will have to live with as uncharted fire activity, water crises, and widespread human footprint collide all-around of the world. Therefore, we advocate for a more proactive approach to wildfire-watershed risk governance in an effort to advance and protect water security. We also argue that there is no easy solution to reducing this risk and that investments in both green (i.e., natural) and grey (i.e., built) infrastructure will be necessary. Further, we propose strategies to combine modern data analytics with existing tools for use by water and land managers worldwide to leverage several decades worth of data and knowledge on post-fire hydrology.


Inferring disease risk genes from sequencing data in multiplex pedigrees through sharing of rare variants.
We previously demonstrated how sharing of rare variants (RVs) in distant affected relatives can be used to identify variants causing a complex and heterogeneous disease. This approach tested whether single RVs were shared by all sequenced affected family members. However, as with other study designs, joint analysis of several RVs (e.g., within genes) is sometimes required to obtain sufficient statistical power. Further, phenocopies can lead to false negatives for some causal RVs if complete sharing among affected is required. Here, we extend our methodology (Rare Variant Sharing, RVS) to address these issues. Specifically, we introduce gene-based analyses, a partial sharing test based on RV sharing probabilities for subsets of affected relatives and a haplotype-based RV definition. RVS also has the desirable feature of not requiring external estimates of variant frequency or control samples, provides functionality to assess and address violations of key assumptions, and is available as open source software for genome-wide analysis. Simulations including phenocopies, based on the families of an oral cleft study, revealed the partial and complete sharing versions of RVS achieved similar statistical power compared with alternative methods (RareIBD and the Gene-Based Segregation Test), and had superior power compared with the pedigree Variant Annotation, Analysis, and Search Tool (pVAAST) linkage statistic. In studies of multiplex cleft families, analysis of rare single nucleotide variants in the exome of 151 affected relatives from 54 families revealed no significant excess sharing in any one gene, but highlighted different patterns of sharing revealed by the complete and partial sharing tests.


Programmed cell death in the Drosophila CNS is ecdysone-regulated and coupled with a specific ecdysone receptor isoform.
At adult emergence, the ventral CNS of Drosophila shows a group of approximately 300 neurons, which are unique in that they express 10-fold higher levels of the A isoform of the ecdysone receptor (EcR-A) than do other central neurons. This expression pattern is established early in metamorphosis and persists throughout the remainder of the pupal stage. Although these cells represent a heterogeneous group of neurons, they all share the same fate of undergoing rapid degeneration after the adult emerges from the pupal case. One prerequisite for this death is the decline of ecdysteroids at the end of metamorphosis. Treatment of flies with 20-hydroxyecdysone blocks the death of the cells, but only if given at least 3 hours before the normal time of degeneration. The correlation of a unique pattern of receptor isoform expression with a particular steroid-regulated fate suggests that variations in the pattern of receptor isoform expression may serve as important switches during development.


Reconstructing three-dimensional hand movements from noninvasive electroencephalographic signals.
It is generally thought that the signal-to-noise ratio, the bandwidth, and the information content of neural data acquired via noninvasive scalp electroencephalography (EEG) are insufficient to extract detailed information about natural, multijoint movements of the upper limb. Here, we challenge this assumption by continuously decoding three-dimensional (3D) hand velocity from neural data acquired from the scalp with 55-channel EEG during a 3D center-out reaching task. To preserve ecological validity, five subjects self-initiated reaches and self-selected targets. Eye movements were controlled so they would not confound the interpretation of the results. With only 34 sensors, the correlation between measured and reconstructed velocity profiles compared reasonably well to that reported by studies that decoded hand kinematics from neural activity acquired intracranially. We subsequently examined the individual contributions of EEG sensors to decoding to find substantial involvement of scalp areas over the sensorimotor cortex contralateral to the reaching hand. Using standardized low-resolution brain electromagnetic tomography (sLORETA), we identified distributed current density sources related to hand velocity in the contralateral precentral gyrus, postcentral gyrus, and inferior parietal lobule. Furthermore, we discovered that movement variability negatively correlated with decoding accuracy, a finding to consider during the development of brain-computer interface systems. Overall, the ability to continuously decode 3D hand velocity from EEG during natural, center-out reaching holds promise for the furtherance of noninvasive neuromotor prostheses for movement-impaired individuals.


Polyneuronal innervation of spiny stellate neurons in cat visual cortex.
Our hypothesis was that spiny stellate neurons in layer 4 of cat visual cortex receive polyneuronal innervation. We characterised the synapses of four likely sources of innervation by three simple criteria: the type of synapse, the target (spine, dendritic shaft), and the area of the presynaptic bouton. The layer 6 pyramids had the smallest boutons and formed asymmetric synapses mainly with the dendritic shaft. The thalamic afferents had the largest boutons and formed asymmetric synapses mainly with spines. The spiny stellates had medium-sized boutons and formed asymmetric synapses mainly with spines. We used these to make a "template" to match against the boutons forming synapses with the spiny stellate dendrite. Of the asymmetric synapses, 45% could have come from layer 6 pyramidal neurons, 28% from spiny stellate neurons, and 6% from thalamic afferents. The remaining 21% of asymmetric synapses could not be accounted for without assuming some additional selectivity of the presynaptic axons. Additional asymmetric synapses may come from a variety of sources, including other cortical neurons and subcortical nuclei such as the claustrum. Of the symmetric synapses, 84% could have been provided by clutch cells, which form large boutons. The remainder, formed by small boutons, probably come from other smooth neurons in layer 4, e.g., neurogliaform and bitufted neurons. Our analysis supports the hypothesis that the spiny stellate receives polyneuronal innervation, perhaps from all the sources of boutons in layer 4. Although layer 4 is the major recipient of thalamic afferents, our results show that they form only a few percent of the synapses of layer 4 spiny stellate neurons.


Does the Presence of Primary Circulating Prostate Cells Imply the Presence of Agressive Prostate Cancer with Early Biochemical Failure: a Comparison with the Walz Nomogram.
To determine the utility of primary circulating prostate cells (CPC) for predicting early biochemical failure after radical prostatectomy for prostate cancer and compare the results with the Walz nomogram. A single centre prospective study of men with prostate cancer treated with radical prostatectomy was conducted between 2004 and 2014. Clinicalpathological details were registered, along with total serum PSA presurgery, Gleason score, extracapsular extension, positive surgical margins, infiltration of lymph nodes, seminal vesicles and pathological stage. Primary circulating prostate cells were obtained using differential gel centrifugation and detected using standard immunocytochemistry with antiPSA. Biochemical failure was defined as a PSA >0.2ng/ml, predictive values were calculated using the Walz nomagram and CPC detection. A total of 285 men participated, of whom 103/285 (36.1%) suffered biochemcial failure; 32/103 (31.1%) within two years of radical prostatectomy. Men with higher Gleason scores, higher pathological stage, infiltration of the surgical margin or prostate capsule and infiltration of seminal vesicles were more likely to undergo biochemical failure. There was a significant increase in the frequency of biochemical failure with increasing number of CPCs detected (p<0.0004 Chi squared for trend) and increasing percent prediction for the Walz nomogram (p<0.0001 Chi squared for trends). The positive predictive value of primary CPC detection, even using a cutoff point of ≥ 4 cells/sample was very low. The detection of primary CPCs in men as a prognostic factor pretreatment fails to identify those at high risk of biochemical failure within two years of curative therapy. This is in keeping with their biological significance, that the majority of them will be eliminated by the primary therapy and thus have no influence on the subsequent clinical history of the patient.