Identification of transient intermediates in the bisphosphatase reaction of rat liver 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase by 31P-NMR spectroscopy.
31P-NMR spectroscopy was used to identify reaction intermediates during catalytic turn-over of the fructose-2,6-bisphosphatase domain (Fru-2,6-P2ase) of the bifunctional enzyme, 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase. When fructose-2,6-bisphosphate (Fru-2,6-P2) was added to the enzyme, the 31P-NMR spectrum showed three resonances in addition to those of free substrate: the phosphohistidine (His-P) intermediate, the C-6 phosphoryl group of fructose-6-phosphate bound to the phosphoenzyme, and phosphate generated by the hydrolysis of substrate. Direct analysis of the alkali-denatured phospho-enzyme intermediate by 1H-31P heteronuclear multiple quantum-filtered coherence spectroscopy confirmed the formation of 3-N-phosphohistidine. Binding of fructose 6-phosphate to the bisphosphatase was detected by a down-field shift and broadening of the C-6 phosphoryl resonance. The down-field shift was greater in the presence of the phosphoenzyme intermediate. Inhibition of Fru-2,6-P2 hydrolysis by fructose 6-phosphate and Fru-2,6-P2 was shown to involve binding of the sugar phosphates to the phosphoenzyme. This study provides new experimental evidence in support of the reaction mechanism of Fru-2,6-P2ase and suggests that the steady-state His-P intermediate exists primarily in the E-P.fructose 6-phosphate complex. These results lay a solid foundation for the use of 31P-NMR magnetization transfer studies to provide an in-depth analysis of the bisphosphatase reaction mechanism.


Antifungal prophylaxis during neutropenia and immunodeficiency.
Fungal infections represent a major source of morbidity and mortality in patients with almost all types of immunodeficiencies. These infections may be nosocomial (aspergillosis) or community acquired (cryptococcosis), or both (candidiasis). Endemic mycoses such as histoplasmosis, coccidioidomycosis, and penicilliosis may infect many immunocompromised hosts in some geographic areas and thereby create major public health problems. With the wide availability of oral azoles, antifungal prophylactic strategies have been extensively developed. However, only a few well-designed studies involving strict criteria have been performed, mostly in patients with hematological malignancies or AIDS. In these situations, the best dose and duration of administration of the antifungal drug often remain to be determined. In high-risk neutropenic or bone marrow transplant patients, fluconazole is effective for the prevention of superficial and/or systemic candidal infections but is not always able to prolong overall survival and potentially selects less susceptible or resistant Candida spp. Primary prophylaxis against aspergillosis remains investigative. At present, no standard general recommendation for primary antifungal prophylaxis can be proposed for AIDS patients or transplant recipients. However, for persistently immunocompromised patients who previously experienced a noncandidal systemic fungal infection, prolonged suppressive antifungal therapy is often indicated to prevent a relapse. Better strategies for controlling immune deficiencies should also help to avoid some potentially life-threatening deep mycoses. When prescribing antifungal prophylaxis, physicians should be aware of the potential emergence of resistant strains, drug-drug interactions, and the cost. Well-designed, randomized, multicenter clinical trials in high-risk immunocompromised hosts are urgently needed to better define how to prevent severe invasive mycoses.


Cerebral blood volume in a rat model of ischemia by MR imaging at 4.7 T.
Perturbation of the cerebral circulation by occlusion of the vertebral arteries and a carotid artery can be visualized by using MR imaging and the intravascular contrast agent Gd-DTPA complexed to albumin. This tracer consistently reduced the T1 relaxation time in the brain and blood. The difference between hemispheres was revealed by less T1 reduction in the occluded hemisphere and by an adjustment in the display contrast of images that revealed the territory of decreased perfusion. These results were confirmed by comparing them with cerebral blood flow using radioactive microspheres and the intravascular blood volume tracer 51Cr-EDTA. This method, combined with high-resolution MR imaging, can be applied to serial noninvasive studies of cerebral blood volume in ischemia and other conditions.


Magnetic resonance electrical properties tomography for small anomalies using boundary conditions: A simulation study.
Magnetic resonance electrical property tomography (MREPT) is an emerging imaging modality using measured B1 maps from magnetic resonance imaging (MRI) to measure a distribution of electric conductivity and permittivity of the subject at the Larmor frequency. Conventional MREPT approaches at single transmit channel system using the Helmholtz equation rely on an assumption that conductivity and permittivity of the subject are locally homogeneous. For small tissue structures and tissue boundaries, in which the assumption of locally homogeneous conductivity and permittivity does not hold, the reconstructed conductivity values deviated from the actual values, so called "Boundary Artifacts." The aim of this study is to propose new reconstruction processes based on time-harmonic Maxwell's equations to reconstruct conductivity for small tissue structures and tissue boundaries. Instead of removing the electric fields from the equations as done in the Helmholtz equation, three key identities of circularly polarized and longitudinal components of electric fields, circularly polarized component of magnetic fields, and electric properties from time-harmonic Maxwell's equations are derived. Based on the three key identities, the proposed reconstruction methods determine conductivity, permittivity, and circularly polarized component and longitudinal component of electric fields using the measured H1+ . In each iterative step, estimated conductivity, permittivity, electric fields, and artifact-free mask region, Ω, where the contribution of the boundary artifacts is small, were updated. Using the estimated values in the artifact-free mask region as boundary conditions, the estimates beyond the mask region were updated. EM simulations were performed on three types of numerical phantoms with very small regions of homogeneous conductivity and permittivity. The performance of the proposed methods was evaluated using the simulated electric and magnetic fields. For the numerical simulation model, the proposed methods significantly reduced the boundary artifacts compared to conventional methods using Helmholtz equations. In addition, previous methods using the Helmholtz equation could measure conductivity of only large anomalies, but the proposed method can measure the conductivity of the small compartments whose size is 2-3 voxels. The proposed approaches are compatible with spatial filtering which can be used to reduce noise. If a good image segmentation is available as a prior information, better initial boundary conditions can be estimated, and thus the proposed approach can be more accurate for small tissue structures. The proposed reconstruction method not only determines electrical properties, but also circularly polarized component and longitudinal component of electric fields using an iterative process. The proposed method can quantitatively detect the conductivity of the small anomalies better than conventional methods.


Blood dyscrasias with carbamazepine and valproate: a pharmacoepidemiological study of 2,228 patients at risk.
The purpose of this study was to determine the occurrence of leukopenia and other blood dyscrasias associated with psychiatric use of carbamazepine and valproate. Rates of WBC counts of 3,000-4,000/mm3 (moderate leukopenia) and < 3,000/mm3 (severe leukopenia), platelet counts of < 100,000/mm3, and hematocrit < 30% were identified among 2,228 treated patients at risk among 11,720 patients admitted to McLean Hospital over 4 years (1989-1993). Patients who received carbamazepine or valproate and had a blood dyscrasia not associated with a relevant medical condition were compared to patients treated with imipramine or desipramine. Of 977 patients treated with carbamazepine, 2.1% experienced leukopenia (16 moderate cases, five severe). Time to 50% risk was 16 days, and recovery occurred within about 6 days after carbamazepine was stopped. For 1,251 patients given valproate, the occurrence of leukopenia was 0.4% (three moderate cases, two severe). The occurrence of leukopenia in 1,031 patients given the tricyclic antidepressants was 0.3% (two moderate cases, one severe). The observed occurrence of moderate leukopenia with carbamazepine was 6.9 and 7.3 times higher than that with valproate and antidepressants, respectively. Severe blood dyscrasias were uncommon in psychiatric patients given carbamazepine and were about as rare with valproate as with imipramine or desipramine. Most important, in this cohort of 2,228 patients exposed to carbamazepine and valproate, there were no life-threatening cases.


A Two-Component Adhesive: Tau Fibrils Arise from a Combination of a Well-Defined Motif and Conformationally Flexible Interactions.
Fibrillar aggregates of Aβ and Tau in the brain are the major hallmarks of Alzheimer's disease. Most Tau fibers have a twisted appearance, but the twist can be variable and even absent. This ambiguity, which has also been associated with different phenotypes of tauopathies, has led to controversial assumptions about fibril constitution, and it is unclear to-date what the molecular causes of this polymorphism are. To tackle this question, we used solid-state NMR strategies providing assignments of non-seeded three-repeat-domain Tau3RD with an inherent heterogeneity. This is in contrast to the general approach to characterize the most homogeneous preparations by construct truncation or intricate seeding protocols. Here, carbon and nitrogen chemical-shift conservation between fibrils revealed invariable secondary-structure properties, however, with inter-monomer interactions variable among samples. Residues with variable amide shifts are localized mostly to N- and C-terminal regions within the rigid beta structure in the repeat region of Tau3RD. By contrast, the hexapeptide motif in repeat R3, a crucial motif for fibril formation, shows strikingly low variability of all NMR parameters: Starting as a nucleation site for monomer-monomer contacts, this six-residue sequence element also turns into a well-defined structural element upon fibril formation. Given the absence of external causes in vitro, the interplay of structurally differently conserved elements in this protein likely reflects an intrinsic property of Tau fibrils.


Plasmodium falciparum Guanylyl Cyclase-Alpha and the Activity of Its Appended P4-ATPase Domain Are Essential for cGMP Synthesis and Blood-Stage Egress.
Guanylyl cyclases (GCs) synthesize cyclic GMP (cGMP) and, together with cyclic nucleotide phosphodiesterases, are responsible for regulating levels of this intracellular messenger which mediates myriad functions across eukaryotes. In malaria parasites (Plasmodium spp), as well as their apicomplexan and ciliate relatives, GCs are associated with a P4-ATPase-like domain in a unique bifunctional configuration. P4-ATPases generate membrane bilayer lipid asymmetry by translocating phospholipids from the outer to the inner leaflet. Here, we investigate the role of Plasmodium falciparum guanylyl cyclase alpha (GCα) and its associated P4-ATPase module, showing that asexual blood-stage parasites lacking both the cyclase and P4-ATPase domains are unable to egress from host erythrocytes. GCα-null parasites cannot synthesize cGMP or mobilize calcium, a cGMP-dependent protein kinase (PKG)-driven requirement for egress. Using chemical complementation with a cGMP analogue and point mutagenesis of a crucial conserved residue within the P4-ATPase domain, we show that P4-ATPase activity is upstream of and linked to cGMP synthesis. Collectively, our results demonstrate that GCα is a critical regulator of PKG and that its associated P4-ATPase domain plays a primary role in generating cGMP for merozoite egress.IMPORTANCE The clinical manifestations of malaria arise due to successive rounds of replication of Plasmodium parasites within red blood cells. Once mature, daughter merozoites are released from infected erythrocytes to invade new cells in a tightly regulated process termed egress. Previous studies have shown that the activation of cyclic GMP (cGMP) signaling is critical for initiating egress. Here, we demonstrate that GCα, a unique bifunctional enzyme, is the sole enzyme responsible for cGMP production during the asexual blood stages of Plasmodium falciparum and is required for the cellular events leading up to merozoite egress. We further demonstrate that in addition to the GC domain, the appended ATPase-like domain of GCα is also involved in cGMP production. Our results highlight the critical role of GCα in cGMP signaling required for orchestrating malaria parasite egress.


Social cohesion through football: a quasi-experimental mixed methods design to evaluate a complex health promotion program.
Social isolation and disengagement fragments local communities. Evidence indicates that refugee families are highly vulnerable to social isolation in their countries of resettlement. Research to identify approaches to best address this is needed. Football United is a program that aims to foster social inclusion and cohesion in areas with high refugee settlement in New South Wales, Australia, through skills and leadership development, mentoring, and the creation of links with local community and corporate leaders and organisations. The Social Cohesion through Football study's broad goal is to examine the implementation of a complex health promotion program, and to analyse the processes involved in program implementation. The study will consider program impact on individual health and wellbeing, social inclusion and cohesion, as well as analyse how the program by necessity interacts and adapts to context during implementation, a concept we refer to as plasticity. The proposed study will be the first prospective cohort impact study to our knowledge to assess the impact of a comprehensive integrated program using football as a vehicle for fostering social inclusion and cohesion in communities with high refugee settlement. A quasi-experimental cohort study design with treatment partitioning involving four study sites. The study employs a 'dose response' model, comparing those with no involvement in the Football United program with those with lower or higher levels of participation. A range of qualitative and quantitative measures will be used in the study. Study participants' emotional well being, resilience, ethnic identity and other group orientation, feelings of social inclusion and belonging will be measured using a survey instrument complemented by relevant data drawn from in-depth interviews, self reporting measures and participant observation. The views of key informants from the program and the wider community will also be solicited. The complexity of the Football United program poses challenges for measurement, and requires the study design to be responsive to the dynamic nature of the program and context. Assessment of change is needed at multiple levels, drawing on mixed methods and multidisciplinary approaches in implementation and evaluation. Attention to these challenges has underpinned the design and methods in the Social Cohesion through Football study, which will use a unique and innovative combination of measures that have not been applied together previously in social inclusion/cohesion and sport and social inclusion/cohesion program research.


Further corroboration of distinct functional features in SCN2A variants causing intellectual disability or epileptic phenotypes.
Deleterious variants in the voltage-gated sodium channel type 2 (Nav1.2) lead to a broad spectrum of phenotypes ranging from benign familial neonatal-infantile epilepsy (BFNIE), severe developmental and epileptic encephalopathy (DEE) and intellectual disability (ID) to autism spectrum disorders (ASD). Yet, the underlying mechanisms are still incompletely understood. To further elucidate the genotype-phenotype correlation of SCN2A variants we investigated the functional effects of six variants representing the phenotypic spectrum by whole-cell patch-clamp studies in transfected HEK293T cells and in-silico structural modeling. The two variants p.L1342P and p.E1803G detected in patients with early onset epileptic encephalopathy (EE) showed profound and complex changes in channel gating, whereas the BFNIE variant p.L1563V exhibited only a small gain of channel function. The three variants identified in ID patients without seizures, p.R937C, p.L611Vfs*35 and p.W1716*, did not produce measurable currents. Homology modeling of the missense variants predicted structural impairments consistent with the electrophysiological findings. Our findings support the hypothesis that complete loss-of-function variants lead to ID without seizures, small gain-of-function variants cause BFNIE and EE variants exhibit variable but profound Nav1.2 gating changes. Moreover, structural modeling was able to predict the severity of the variant impact, supporting a potential role of structural modeling as a prognostic tool. Our study on the functional consequences of SCN2A variants causing the distinct phenotypes of EE, BFNIE and ID contributes to the elucidation of mechanisms underlying the broad phenotypic variability reported for SCN2A variants.


Mobility of histones on the chromosome of simian virus 40.
Linear simian virus 40 (SV40) chromosomes were prepared by Eco R1 nuclease cleavage of the circular SV40 chromosomes released from virions with dithiothreitol at pH 9,8. Chromatin-DNA hybrids were constructed with segments of 3H-labeled, naked SV40 DNA covalently joined via the Eco R1-generated cohesive ends to segments of linear SV40 chromosome. Upon incubation of chromatin-DNA hybrids at 37 degrees C and moderate ionic strength, histones migrated onto the labeled DNA while retaining the nucleosome structure. This was shown first, by the pattern of micrococcal nuclease digestion of labeled DNA; second by nitrocellulose filter binding of labeled DNA after redigestion of the chromatin-DNA hybrids with Eco R1; and third, by examination of chromatin-DNA hybrids in the electron microscope. Migration was slow, being apparent after several hours. Parallel experiments in which naked DNA and chromosomes were mixed without joining showed no transfer of nucleosomal histones between DNA molecules. The kinetics of Eco R1 cleavage of the DNA in virion-derived SV40 chromosomes are also consistent with the notion that nucleosomal histones, in the absence of other proteins, can move on DNA.


United Kingdom: coronary and structural heart interventions from 2010 to 2015.
In the United Kingdom, a clinical data set is completed for every patient undergoing coronary intervention and certain structural interventions, and sent to central servers in the National Institute for Cardiovascular Outcomes Research (NICOR) on behalf of the British Cardiovascular Intervention Society (BCIS). These data are linked to the national mortality register. In addition, data are obtained about the structure of healthcare provision using an annual survey. Analyses of these data are provided for different audiences in several formats. Public reports of individual consultant operator activity and risk-adjusted outcomes from percutaneous coronary intervention (PCI) have also been produced annually since 2012. Transcatheter aortic valve implantation (TAVI) has been performed since 2007. Over 2,000 cases were performed in 2015, giving a rate of 30 per million population. Complications to discharge have fallen as case mix has changed and technologies improved. While the mean age has remained about 81 years, the logistic EuroSCORE of patients treated by TAVI has fallen from about 22 in 2010 to 18 in 2015. Tracked 30-day mortality was 3.7% in 2014. Left atrial appendage occlusion and patent foramen ovale (PFO) closure for stroke, and the use of the MitraClip® system (Abbott Vascular, Santa Clara, CA, USA) for mitral regurgitation have been funded through a process called "commissioning through evaluation".


Transcriptomic characterization of Caecomyces churrovis: a novel, non-rhizoid-forming lignocellulolytic anaerobic fungus.
Anaerobic gut fungi are the primary colonizers of plant material in the rumen microbiome, but are poorly studied due to a lack of characterized isolates. While most genera of gut fungi form extensive rhizoidal networks, which likely participate in mechanical disruption of plant cell walls, fungi within the Caecomyces genus do not possess these rhizoids. Here, we describe a novel fungal isolate, Caecomyces churrovis, which forms spherical sporangia with a limited rhizoidal network yet secretes a diverse set of carbohydrate active enzymes (CAZymes) for plant cell wall hydrolysis. Despite lacking an extensive rhizoidal system, C. churrovis is capable of growth on fibrous substrates like switchgrass, reed canary grass, and corn stover, although faster growth is observed on soluble sugars. Gut fungi have been shown to use enzyme complexes (fungal cellulosomes) in which CAZymes bind to non-catalytic scaffoldins to improve biomass degradation efficiency. However, transcriptomic analysis and enzyme activity assays reveal that C. churrovis relies more on free enzymes compared to other gut fungal isolates. Only 15% of CAZyme transcripts contain non-catalytic dockerin domains in C. churrovis, compared to 30% in rhizoid-forming fungi. Furthermore, C. churrovis is enriched in GH43 enzymes that provide complementary hemicellulose degrading activities, suggesting that a wider variety of these activities are required to degrade plant biomass in the absence of an extensive fungal rhizoid network. Overall, molecular characterization of a non-rhizoid-forming anaerobic fungus fills a gap in understanding the roles of CAZyme abundance and associated degradation mechanisms during lignocellulose breakdown within the rumen microbiome.


Nematodes in Hoplerytrinus unitaeniatus, Hoplias malabaricus and Pygocentrus nattereri (pisces characiformes) in Marajó Island, Brazil.
The aim of this study was to evaluate the tegument, musculature and mesentery of 102 specimens of Hoplerytrinus unitaeniatus, 104 of Hoplias malabaricus and 101 of Pygocentrus nattereri, from Arari Lake, Marajó Island, State of Pará, Brazil. Were identified the nematodes Contracaecum sp., Eustrongylides sp. and Procamallanus sp. Contracaecum sp. was the most prevalent, with rates of 84.31% (H. unitaeniatus), 95.19% (H. malabaricus), and 89.11% (P. nattereri). The highest prevalences of Eustrongylides sp. occurred in H. unitaeniatus (56.86%) and H. malabaricus (53.84%). Procamallanus sp. was only collected in the mesentery. Specimens of Eustrongylides sp. collected from the musculature were 91.9% of its population. Among the nematodes found in the mesentery, 98.34% were Contracaecum sp. with a mean intensity (MI) of 7.92 ± 8.11 (H. unitaeniatus), 8.49 ± 8.34 (H. malabaricus) and 7 ± 6.40 (P. nattereri). Contracaecum sp. presented the highest MI (8.49 ± 8.34) and mean abundance (8.09 ± 8.34). The highest MI values were observed in the mesentery. Eustrongylides sp. presented MI of 2.65 ± 3.21 (H. unitaeniatus), 3.41 ± 3.27 (H. malabaricus) and 2.17 ± 1.18 (P. nattereri). Nematodes with zoonotic potential that were found with high prevalence, shows the importance of actions by the health authorities.


Stride length of elderly patients with lumbar spinal stenosis: Multi-center study using the Two-Step test.
Short stride length is one of clinical symptoms associated with lumbar spinal stenosis (LSS). Short stride is a risk factor for falls; therefore, identification of factors associated with short stride is critical for fall prevention in LSS patients. Although the Two-Step test can conveniently assess maximal stride length, it has not become widely used; therefore, its data are limited. We identified the potential factors associated with short stride of elderly LSS patients using Two-Step test. Clinical data of patients aged >65 years who planned to undergo surgery for LSS were prospectively collected at multiple institutions. Patients were assessed with the Two-Step test and Timed Up-and-Go Test prior to surgery; 357 consecutive patients were enrolled. We determined the cut-off value of the Two-Step test score for short stride, referring to the Timed Up-and-Go Test score of 13.5 s, used to indicate high risk of falls in elderly individuals. Logistic regression model was constructed to identify factors associated with short stride. The Two-Step test score showed moderate-to-strong inverse correlation with that of Timed Up-and-Go Test (r = -0.65, p < 0.001). Using the tentative Two-Step test cut-off value (0.93) for short stride, multivariable analysis showed that age ≥80 years (OR = 2.3, 95% CI:1.1-4.8), a score of <60 for lumbar function in Japanese Orthopedic Association Back Pain Evaluation Questionnaire (OR = 2.7, 95% CI:1.5-4.7), motor deficit (OR = 2.7, 95% CI:1.2-6.1), and sagittal vertical axis ≥50 mm (OR = 2.1, 95% CI:1.2-3.5) were factors significantly associated with short stride in elderly patients with LSS. Using the Two-Step test, we found that 80 years old and over, lumbar dysfunction, motor deficit of the lower extremities, and forward-bent posture were associated with short stride in LSS patients. Therefore, elderly LSS patients with these conditions may have a higher risk for falls.


Protein kinase C remains functionally active during TPA induced neuronal differentiation of SH-SY5Y human neuroblastoma cells.
SH-SY5Y human neuroblastoma cells can be induced to differentiate into a neuronal phenotype by treatment with 12-O-tetradecanoylphorbol-13-acetate (TPA). In other cell systems, TPA treatment frequently leads to down-regulation of protein kinase C (PKC). However, we now report that TPA-treated and non-treated SH-SY5Y cells express PKC-alpha, but not PKC-beta and PKC-gamma, mRNA. Furthermore, only a slight down-regulation of the PKC-alpha protein could be seen during prolonged treatment with 16 nM TPA, the concentration giving optimal differentiation. In contrast, a higher concentration of TPA (1.6 microM) results in a poor neuronal differentiation and a complete down-regulation of PKC-alpha. PKC-alpha was rapidly translocated to the particulate fraction and remained membrane bound for at least 4 days during treatment with 16 nM TPA. In such cells a sustained increased level of the phosphorylated form of a 80,000 Dalton PKC-substrate was found. In addition to this sustained augmented phosphorylation, administration of fresh TPA at day 4 caused a small but reproducible further increased level of phosphorylated substrate. When the PKC activity was measured by the histone phosphorylation assay a substantial fraction of the initial enzyme activity could still be detected after 4 days of TPA treatment. Taken together, the data demonstrate that PKC remains functionally active during TPA induced differentiation of SH-SY5Y cells, which may suggest a continuous role for the enzyme during the differentiation process.


Models of cooperativity in protein folding.
What is the basis for the two-state cooperativity of protein folding? Since the 1950s, three main models have been put forward. 1. In 'helix-coil' theory, cooperativity is due to local interactions among near neighbours in the sequence. Helix-coil cooperativity is probably not the principal basis for the folding of globular proteins because it is not two-state, the forces are weak, it does not account for sheet proteins, and there is no evidence that helix formation precedes the formation of a hydrophobic core in the following pathways. 2. In the 'sidechain packing' model, cooperativity is attributed to the jigsaw-puzzle-like complementary fits of sidechains. This too is probably not the basis of folding cooperativity because exact models and experiments on homopolymers with sidechains give no evidence that sidechain freezing is two-state, sidechain complementarities in proteins are only weak trends, and the molten globule model predicted by this model is far more native-like than experiments indicate. 3. In the 'hydrophobic core collapse' model, cooperativity is due to the assembly of non-polar residues into a good core. Exact model studies show that this model gives two-state behaviour for some sequences of hydrophobic and polar monomers. It is based on strong forces. There is considerable experimental evidence for the kinetics this model predicts: the development of hydrophobic clusters and cores is concurrent with secondary structure formation. It predicts compact denatured states with sizes and degrees of disorder that are in reasonable agreement with experiments.


Structure and expression pattern of Oct4 gene are conserved in vole Microtus rossiaemeridionalis.
Oct4 is a POU-domain transcriptional factor which is essential for maintaining pluripotency in several mammalian species. The mouse, human, and bovine Oct4 orthologs display a high conservation of nucleotide sequence and genomic organization. Here we report an isolation of a common vole (Microtus rossiaemeridionalis) Oct4 ortholog. Organization and exon-intron structure of vole Oct4 gene are similar to the gene organization in other mammalian species. It consists of five exons and a regulatory region including the minimal promoter, proximal and distal enhancers. Promoter and regulatory regions of the vole Oct4 gene also display a high similarity to the corresponding regions of Oct4 in other mammalian species, and are active during the transient transfection within luciferase reporter constructs into mouse P19 embryonic carcinoma cells and TG-2a embryonic stem cells. The vole Oct4 gene expression is detectable starting from the morula stage and until day 17 of embryonic development. Genomic organization of this gene and its intron-exon structure in vole are identical to those in all previously studied species: it comprises five exons and the regulatory region containing several conserved elements. The activity of the Oct4 gene in vole, as well as in mouse, is confined only to pluripotent cells.


Possible roles of spontaneous waves and dendritic growth for retinal receptive field development.
Several models of cortical development postulate that a Hebbian process fed by spontaneous activity amplifies orientation biases occurring randomly in early wiring, to form orientation selectivity. These models are not applicable to the development of retinal orientation selectivity, since they neglect the polarization of the retina's poorly branched early dendritic trees and the wavelike organization of the retina's early noise. There is now evidence that dendritic polarization and spontaneous waves are key in the development of retinal receptive fields. When models of cortical development are modified to take these factors into account, one obtains a model of retinal development in which early dendritic polarization is the seed of orientation selectivity, while the spatial extent of spontaneous waves controls the spatial profile of receptive fields and their tendency to be isotropic.


Thermal dosimetry characteristics of deep regional heating of non-muscle invasive bladder cancer.
The aim of this paper is to report thermal dosimetry characteristics of external deep regional pelvic hyperthermia combined with intravesical mitomycin C (MMC) for treating bladder cancer following transurethral resection of bladder tumour, and to use thermal data to evaluate reliability of delivering the prescribed hyperthermia dose to bladder tissue. A total of 14 patients were treated with MMC and deep regional hyperthermia (BSD-2000, Sigma Ellipse or Sigma 60). The hyperthermia objective was 42° ± 2 °C to bladder tissue for ≥40 min per treatment. Temperatures were monitored with thermistor probes and recorded values were used to calculate thermal dose and evaluate treatment. Anatomical characteristics were examined for possible correlations with heating. Combined with BSD-2000 standard treatment planning and patient feedback, real-time temperature monitoring allowed thermal steering of heat sufficient to attain the prescribed thermal dose to bladder tissue within patient tolerance in 91.6% of treatments. Mean treatment time for bladder tissue >40 °C was 61.9 ± 11.4 min and mean thermal dose was 21.3 ± 16.5 CEM43. Average thermal doses obtained in normal tissues were 1.6 ± 1.2 CEM43 for the rectum and 0.8 ± 1.3 CEM43 in superficial normal tissues. No significant correlation was seen between patient anatomical characteristics and thermal dose achieved in bladder tissue. This study demonstrates that a hyperthermia prescription of 42° ± 2 °C for 40-60 min can be delivered safely to bladder tissue with external radiofrequency phased array applicators for a typical range of patient sizes. Using the available thermometry and treatment planning, the BSD-2000 hyperthermia system was shown to be an effective method of focusing heat regionally around the bladder with good patient tolerance.


Enhancing spatial resolution in digital holographic microscopy by biprism structured illumination.
A novel and efficient architecture of a structured-illumination digital holographic microscope (DHM) is presented. As the DHM operates at the diffraction limit, its spatial resolution on label-free imaging of transparent samples is improved by illuminating the sample with a structured illumination produced by a Fresnel's biprism. The theoretical analysis of the method forecasts a twofold improvement of the spatial resolution. The proposed method requires only two images to improve the spatial resolution, which eases the process of unmixing the high-resolution components by means of an unknown phase-shift procedure. Numerical modeling and experimental results validate the theoretical findings.


Diagnostic ionizing radiation exposure in a population-based cohort of patients with inflammatory bowel disease.
For diagnosis, assessing disease activity, complications and extraintestinal manifestations, and monitoring response to therapy, patients with inflammatory bowel disease undergo many radiological studies employing ionizing radiation. However, the extent of radiation exposure in these patients is unknown. A population-based inception cohort of 215 patients with inflammatory bowel disease from Olmsted County, Minnesota, diagnosed between 1990 and 2001, was identified. The total effective dose of diagnostic ionizing radiation was estimated for each patient. Linear regression was used to assess the median total effective dose since symptom onset. The number of patients with Crohn's disease and ulcerative colitis was 103 and 112, with a mean age at diagnosis of 38.6 and 39.4 yr, respectively. Mean follow-up was 8.9 yr for Crohn's disease and 9.0 yr for ulcerative colitis. Median total effective dose for Crohn's disease was 26.6 millisieverts (mSv) (range, 0-279) versus 10.5 mSv (range, 0-251) for ulcerative colitis (P < 0.001). Computed tomography accounted for 51% and 40% of total effective dose, respectively. Patients with Crohn's disease had 2.46 times higher total effective dose than ulcerative colitis patients (P= 0.001), adjusting for duration of disease. Annualizing our data, the radiation exposure in the inflammatory bowel disease population was equivalent to the average annual background radiation dose from naturally occurring sources in the U.S. (3.0 mSv). However, a subset of patients had substantially higher doses. The development of imaging management guidelines to minimize radiation dose, dose-reduction techniques in computed tomography, and faster, more robust magnetic resonance techniques are warranted.


Candidate sampling for neuron reconstruction from anisotropic electron microscopy volumes.
The automatic reconstruction of neurons from stacks of electron microscopy sections is an important computer vision problem in neuroscience. Recent advances are based on a two step approach: First, a set of possible 2D neuron candidates is generated for each section independently based on membrane predictions of a local classifier. Second, the candidates of all sections of the stack are fed to a neuron tracker that selects and connects them in 3D to yield a reconstruction. The accuracy of the result is currently limited by the quality of the generated candidates. In this paper, we propose to replace the heuristic set of candidates used in previous methods with samples drawn from a conditional random field (CRF) that is trained to label sections of neural tissue. We show on a stack of Drosophila melanogaster neural tissue that neuron candidates generated with our method produce 30% less reconstruction errors than current candidate generation methods. Two properties of our CRF are crucial for the accuracy and applicability of our method: (1) The CRF models the orientation of membranes to produce more plausible neuron candidates. (2) The interactions in the CRF are restricted to form a bipartite graph, which allows a great sampling speed-up without loss of accuracy.


Estimating HIV-1 fitness characteristics from cross-sectional genotype data.
Despite the success of highly active antiretroviral therapy (HAART) in the management of human immunodeficiency virus (HIV)-1 infection, virological failure due to drug resistance development remains a major challenge. Resistant mutants display reduced drug susceptibilities, but in the absence of drug, they generally have a lower fitness than the wild type, owing to a mutation-incurred cost. The interaction between these fitness costs and drug resistance dictates the appearance of mutants and influences viral suppression and therapeutic success. Assessing in vivo viral fitness is a challenging task and yet one that has significant clinical relevance. Here, we present a new computational modelling approach for estimating viral fitness that relies on common sparse cross-sectional clinical data by combining statistical approaches to learn drug-specific mutational pathways and resistance factors with viral dynamics models to represent the host-virus interaction and actions of drug mechanistically. We estimate in vivo fitness characteristics of mutant genotypes for two antiretroviral drugs, the reverse transcriptase inhibitor zidovudine (ZDV) and the protease inhibitor indinavir (IDV). Well-known features of HIV-1 fitness landscapes are recovered, both in the absence and presence of drugs. We quantify the complex interplay between fitness costs and resistance by computing selective advantages for different mutants. Our approach extends naturally to multiple drugs and we illustrate this by simulating a dual therapy with ZDV and IDV to assess therapy failure. The combined statistical and dynamical modelling approach may help in dissecting the effects of fitness costs and resistance with the ultimate aim of assisting the choice of salvage therapies after treatment failure.


Air-sea waves from the explosion of krakatoa.
The distant sea disturbances which followed the explosion of Krakatoa are correlated with recently discovered atmospheric acoustic and gravity modes having the same phase velocity as long waves on the ocean. The atmospheric waves jumped over the land barriers and reexcited the sea waves with amplitudes exceeding the hydrostatic values. An explosion of 100 to 150 megatons would be required to duplicate the Krakatoa atmosphericpressure pulse.


Injury-induced asymmetric cell death as a driving force for head regeneration in Hydra.
The freshwater Hydra polyp provides a unique model system to decipher the mechanisms underlying adult regeneration. Indeed, a single cut initiates two distinct regenerative processes, foot regeneration on one side and head regeneration on the other side, the latter relying on the rapid formation of a local head organizer. Two aspects are discussed here: the asymmetric cellular remodeling induced by mid-gastric bisection and the signaling events that trigger head organizer formation. In head-regenerating tips (but not in foot ones), a wave of cell death takes place immediately, leading the apoptotic cells to transiently release Wnt3 and activate the β-catenin pathway in the neighboring cycling cells to push them through mitosis. This process, which mimics the apoptosis-induced compensatory proliferation process deciphered in Drosophila larvae regenerating their discs, likely corresponds to an evolutionarily conserved mechanism, also at work in Xenopus tadpoles regenerating their tail or mice regenerating their skin or liver. How is this process generated in Hydra? Several studies pointed to the necessary activation of the extracellular signal-regulated kinase (ERK) 1-2 and mitogen-activated protein kinase (MAPK) pathways during early head regeneration. Indeed inhibition of ERK 1-2 or knockdown of RSK, cAMP response element-binding protein (CREB), and CREB-binding protein (CBP) prevent injury-induced apoptosis and head regeneration. The current scenario involves an asymmetric activation of the MAPK/CREB pathway to trigger injury-induced apoptosis in the interstitial cells and in the epithelial cells a CREB/CBP-dependent transcriptional activation of early genes essential for head-organizing activity as wnt3, HyBra1, and prdl-a. The question now is how bisection in the rather uniform central region of the polyp can generate this immediately asymmetric signaling.


A pilot study evaluating changes in pancreatic lipase immunoreactivity concentrations in canines treated with L-asparaginase (ASNase), vincristine, or both for lymphoma.
L-asparaginase (ASNase) is a common chemotherapy agent for the treatment of lymphoid malignancies. L-asparaginase has been reported to cause clinical pancreatitis in both humans and canines. Canine pancreatic lipase immunoreactivity (cPLI) is now a common diagnostic tool for evaluating pancreatitis in dogs. A total of 52 dogs were enrolled into this study. Canine pancreatic lipase immunoreactivity (cPLI) concentrations were evaluated before and after administration of ASNase, vincristine, or both. All dogs enrolled in the study were evaluated for signs compatible with clinical pancreatitis. No dogs receiving ASNase alone showed evidence of clinical pancreatitis after administration. Also, there was no statistically significant change in cPLI concentrations before or after treatment. Fourteen percent of dogs that received both vincristine and ASNase concurrently had elevated concentrations of cPLI after treatment. Of the 11 dogs with clinical signs compatible with pancreatitis after any chemotherapy treatment, no dog had a cPLI concentration > 400 microg/dL. In conclusion, ASNase did not cause clinical pancreatitis in this cohort of dogs but larger sample sizes are required to further validate this data.


Spatial convergence of distant cortical regions during folding explains why arteries do not cross the sylvian fissure.
Cortical folding places regions that are separated by a large distance along the cortical surface in close proximity. This process is not homogeneous; regions such as the insular opercula have a much higher cortical surface distance (CSD) to euclidean distance (ED) than others. Here the authors explore the hypothesis that in the folded brain the CSD, and not the ED, determines regions of common irrigation, because this measure corresponds more closely with the distance along the prefolded brain, where the subarachnoid arterial vascular network starts forming. The authors defined a convergence index that compared the ED to the CSD and applied it to the cortical surface reconstruction of an average brain. They then compared cortical convergence to the irrigation patterns of major sulci and fissures of the brain, by assessing whether these structures were crossed or not crossed by arterial vessels in 20 fixed hemispheres. The regions of highest convergence (top 1%) were clustered around the sylvian fissure, which is the only brain depression with high convergence values along its edges. Arterial crossings were commonly observed in every major sulcus of the brain, with the exception of the sylvian fissure, constituting a highly significant difference (p < 10-4). Arteries do not cross regions of high convergence. In the adult brain the CSD, rather than the ED, predicts the regional irrigation pattern. The distant origin of the frontal and temporal lobes creates a region of high cortical convergence, which explains why arteries do not cross the sylvian fissure.


Listeria monocytogenes contamination of ready-to-eat foods and the risk for human health in the EU.
Food safety criteria for Listeria monocytogenes in ready-to-eat (RTE) foods have been applied from 2006 onwards (Commission Regulation (EC) 2073/2005). Still, human invasive listeriosis was reported to increase over the period 2009-2013 in the European Union and European Economic Area (EU/EEA). Time series analysis for the 2008-2015 period in the EU/EEA indicated an increasing trend of the monthly notified incidence rate of confirmed human invasive listeriosis of the over 75 age groups and female age group between 25 and 44 years old (probably related to pregnancies). A conceptual model was used to identify factors in the food chain as potential drivers for L. monocytogenes contamination of RTE foods and listeriosis. Factors were related to the host (i. population size of the elderly and/or susceptible people; ii. underlying condition rate), the food (iii. L. monocytogenes prevalence in RTE food at retail; iv. L. monocytogenes concentration in RTE food at retail; v. storage conditions after retail; vi. consumption), the national surveillance systems (vii. improved surveillance), and/or the bacterium (viii. virulence). Factors considered likely to be responsible for the increasing trend in cases are the increased population size of the elderly and susceptible population except for the 25-44 female age group. For the increased incidence rates and cases, the likely factor is the increased proportion of susceptible persons in the age groups over 45 years old for both genders. Quantitative modelling suggests that more than 90% of invasive listeriosis is caused by ingestion of RTE food containing > 2,000 colony forming units (CFU)/g, and that one-third of cases are due to growth in the consumer phase. Awareness should be increased among stakeholders, especially in relation to susceptible risk groups. Innovative methodologies including whole genome sequencing (WGS) for strain identification and monitoring of trends are recommended.


Social class, political powere, and the state and their implications in medicine.
This three-part article presents an analysis of the distribution of power and of the nature of the state in Western industrialized societies, and details their implications in medicine. Part I presents a critique of contemporary theories of the Western system of power; discusses the countervailing pluralist and power elite theories, as well as those of bureaucratic and professional control; and concludes with an examination of the Marxist theories of economic determinism, structural determinism, and corporate statism. Part II presents a Marxist theory of the role, nature, and characteristics of state intervention. Part III focuses on the mode of that intervention and the reasons for its growth, with an added analysis of the attributes of state intervention in the health sector and of the dialectical relationship between its growth and the current fiscal crisis of the state. In all three parts, the focus is on Western European countries and on North America, with many examples and categories drawn from the area of medicine.


Preferential binding of the histone (H3-H4)2 tetramer by NAP1 is mediated by the amino-terminal histone tails.
The yeast nucleosome assembly protein 1 (yNAP1) participates in many diverse activities, such as the assembly of newly synthesized DNA into chromatin and the rearrangement of nucleosomes during transcriptional activation. yNAP1 does not require ATP hydrolysis to perform these functions and is a valuable tool for in vitro chromatin assembly. Using recombinant histone complexes, we show that yNAP1 has a preference for binding the (H3-H4)2 tetramer over the (H2A-H2B) dimer. We find that the loss of the histone tails abrogates this preference for H3 and H4, and we demonstrate a direct interaction between yNAP1 and the amino-terminal tails of H3 and H4. yNAP1 binds to one histone fold domain, thus specifying the stoichiometry of the complexes formed with the histone dimer and tetramer. Finally, we provide evidence that the acidic carboxyl-terminal region of yNAP1, although dispensable for nucleosome assembly in vitro, contributes to binding via structure-independent electrostatic interactions. Our results are consistent with recent mechanistic investigations of NAP1 and expand our understanding of the histone chaperone family of assembly factors.


Chicory (Cichorium intybus L.) polysaccharides attenuate high-fat diet induced non-alcoholic fatty liver disease via AMPK activation.
Chicory polysaccharides (CP) from Cichorium intybus L. roots were extracted and fractionated to isolate two novel polysaccharide fractions, CP-1 and CP-2. CP-1 is a heteropolysaccharide that is mainly composed of sorbin, glucose, fructose, and glucitol at a molar ratio of 1.00:5.58:13.97:10.32. The molecular weight of CP-1 was 8511.4 Da. The hepatoprotective effect of CP-1 was investigated in a rat model of high-fat diet induced non-alcoholic fatty liver disease (NAFLD) rats. Results indicated that the oral administration of CP-1 significantly decreased body weight and liver index in NAFLD rats. CP-1 also significantly increased serum levels of SOD and HDLC, and decreased the levels of ALT, AST, TG, TC, LDL-C, GLU, ALP, LDH, and MDA in NAFLD rats. Meanwhile, CP-1 effectively decreased MDA, TC, and TG, and increased SOD and T-AOC in the livers of NAFLD rats. Furthermore, CP-1 also increased the hepatic expression of p-AMPKα, ATGL, CPT-1, and p-ACC, and reduced the hepatic expression of ACC, FAS, and SCD-1. Moreover, histopathological examination of the livers showed that CP-1 significantly ameliorated the symptoms of NAFLD rats. Therefore, CP-1 significantly attenuated the high-fat diet-induced NAFLD in rats via AMPK activation.


Type I Interferon Receptor Expression in Human Pancreatic and Periampullary Cancer Tissue.
Interferons (IFNs) have several anticancer mechanisms. A number of clinical trials have been conducted regarding adjuvant IFN-α therapy in pancreatic cancer. Type I IFNs exert their effect via the type I IFN receptor (IFNAR-1, IFNAR-2c). The aims of the present study were to determine the type I IFN receptor expression in pancreatic and periampullary cancer tissues and to study its relation with clinicopathological factors. Receptor expression was determined by immunohistochemistry in paraffin-embedded cancer tissue of 47 pancreatic and 54 periampullary cancer patients. The results demonstrated that 91.5% of the pancreatic tumors and 88.9% of the periampullary tumors showed expression of IFNAR-1, of which 23.4% and 13.0% were strongly positive, respectively. Regarding IFNAR-2c expression, 68.1% of the pancreatic tumors and 68.5% of the periampullary tumors were positive, of which 4.3% of the pancreatic tumors and none of the periampullary tumors had a strong expression. No statistically significant associations were found between type I IFN receptor expression and clinicopathological factors or survival. Type I IFN receptors are expressed in pancreatic and periampullary cancer tissues although with great intertumoral and intratumoral variability. A small proportion of both tumors showed a strong expression of the IFNAR-1; only a very small percentage of the pancreatic tumors showed strong expression of the IFNAR-2c.


Complete nutrient content of four species of commercially available feeder insects fed enhanced diets during growth.
Commercially raised feeder insects used to feed captive insectivores are a good source of many nutrients but are deficient in several key nutrients. Current methods used to supplement insects include dusting and gut-loading. Here, we report on the nutrient composition of four species of commercially raised feeder insects fed a special diet to enhance their nutrient content. Crickets, mealworms, superworms, and waxworms were analyzed for moisture, crude protein, fat, ash, acid detergent fiber, total dietary fiber, minerals, amino acids, fatty acids, vitamins, taurine, carotenoids, inositol, and cholesterol. All four species contained enhanced levels of vitamin E and omega 3 fatty acids when compared to previously published data for these species. Crickets, superworms, and mealworms contained β-carotene although using standard conversion factors only crickets and superworms would likely contain sufficient vitamin A activity for most species of insectivores. Waxworms did not contain any detectable β-carotene but did contain zeaxanthin which they likely converted from dietary β-carotene. All four species contained significant amounts of both inositol and cholesterol. Like previous reports all insects were a poor source of calcium and only superworms contained vitamin D above the limit of detection. When compared to the nutrient requirements as established by the NRC for growing rats or poultry, these species were good sources of most other nutrients although the high fat and low moisture content of both waxworms and superworms means when corrected for energy density these two species were deficient in more nutrients than crickets or mealworms. These data show the value of modifying the diet of commercially available insects as they are growing to enhance their nutrient content. They also suggest that for most insectivores properly supplemented lower fat insects such as crickets, or smaller mealworms should form the bulk of the diet.


Diversification of African tree frogs (genus Leptopelis) in the highlands of Ethiopia.
The frog genus Leptopelis is composed of ~50 species that occur across sub-Saharan Africa. The majority of these frogs are typically arboreal; however, a few species have evolved a fossorial lifestyle. Most species inhabit lowland forests, but a few species have adapted to high elevations. Five species of Leptopelis occupy the Ethiopian highlands and provide a good opportunity to study the evolutionary transition from an arboreal to a fossorial lifestyle, as well as the diversification in this biodiversity hot spot. We sequenced 14 nuclear and three mitochondrial genes, and generated thousands of SNPs from ddRAD sequencing to study the evolutionary relationships of Ethiopian Leptopelis. The five species of highland Leptopelis form a monophyletic group, which diversified during the late Miocene and Pliocene. We found strong population structure in the fossorial species L. gramineus, with levels of genetic differentiation between populations similar to those found between arboreal species. This could indicate that L. gramineus is a complex of cryptic species. We propose that after the original colonization of the Ethiopian highlands by the ancestor of the L. gramineus group, episodes of vicariance fragmented the ancestral populations of this group. We also report the re-evolution of arboreality in L. susanae, which evolved from a fossorial ancestor, a rare ecological switch in frogs that had previously been reported only once.


Strong stretching theory for pH-responsive polyelectrolyte brushes in large salt concentrations.
In this paper, we develop a theory for describing the thermodynamics, configuration, and electrostatics of strongly-stretched, pH-responsive polyelectrolyte (PE) brushes in the presence of large salt concentrations. The aim of the paper, therefore, is to study the properties of a PE brush in a salt concentration regime (namely, large concentrations of several molars) that has been hitherto unexplored theoretically in the context of PE brushes but can be routinely encountered in molecular scale simulations of the problem. The brushes are modelled using our recently developed augmented Strong Stretching Theory (SST), while the effect of the presence of the large salt concentration is accounted for by including the contributions of three different types of non-Poisson-Boltzmann (non-PB) effects to the free energy description of the PE brush induced electric double layer (EDL). These non-PB effects are ionic non-mean-field ion-ion correlations, solvent polarization, and the finite size effect of the ions and water dipoles. We study the individual influences of these different effects and show that the ion-ion correlations and solvent polarization effect reduce the brush height which consequently enhances the monomer density and leads to an electrostatic potential distribution of the brush induced EDL that has a larger magnitude at near-wall locations and becomes zero at shorter distances from the wall. The finite size effect, on the other hand, increases the brush height and therefore, weakens the monomer density and leads to a smaller near-wall magnitude of the EDL potential that becomes zero at larger distances from the wall. Eventually, we consider the impact of all the three non-PB effects simultaneously and show that the ion-ion correlations and solvent polarization effect dominate the size effects and dictate the overall brush configuration and the EDL electrostatics. We also point out that the influence of all the three non-PB effects becomes the largest for a larger salt concentration and a smaller bulk pH. Finally, we compare our theoretical predictions with those obtained from our recently developed all-atom MD simulation model and obtain an excellent match.


Multifocal pancreatic ductal adenocarcinomas concomitant with intraductal papillary mucinous neoplasms of the pancreas detected by intraoperative pancreatic juice cytology. A case report.
Intraductal papillary mucinous neoplasms (IPMNs) of the pancreas have been detected with increasing frequency as a result of the progression of diagnostic modalities. Recently, invasive ductal carcinoma of the pancreas concomitant with IPMNs has been the focus of attention. We report the case of a 57-year-old man with multifocal ductal carcinomas of the pancreas concomitant with IPMNs detected by intraoperative cytology. During a follow-up for branch duct IPMNs, a stenotic lesion of the main duct in the pancreatic body was found by ERCP, and brush cytology of the stenosis revealed an adenocarcinoma. A distal pancreatectomy was proposed; however, intraoperative pancreatic juice cytology from the pancreatic head also revealed adenocarcinoma, and a total pancreatectomy was finally carried out. Pathological examination of the resected specimen showed multifocal ductal carcinomas and IPMNs in the distal pancreas, and invasive ductal carcinoma in the pancreatic head which had not been detected by preoperative imaging studies. Surgeons should be aware of the possibility of multifocal carcinomas in patients with concomitant IPMNs. Intraoperative pancreatic juice cytology should always be performed in order to confirm the absence of carcinoma in the pancreas to be left in place after planned resection.


Process evaluation of a social support platform 'Inlife' for caregivers of people with dementia.
Informal caregivers of persons with dementia have an increased risk of facing social isolation. Online social media interventions might offer a new opportunity to increase access to social support. An online social support platform, 'Inlife', was developed and launched in the Netherlands to enhance social support, positive interactions and information sharing in informal support networks. A process evaluation was performed to evaluate the internal and external validity of the Inlife intervention. Implementation, sampling and intervention quality were evaluated by both qualitative and quantitative methods. Analyses were performed using descriptive statistics and inductive content analysis. Analyses were conducted following participants' completion of the intervention after 16 weeks. The overall participation rate in the study was 27% (96/351). The Inlife intervention was generally well-received by the primary caregivers. Inlife facilitated empowerment, openness, involvement, and efficient care organization. Still, adherence was not optimal for all Inlife users. Determinants for Inlife use were identified on the level of the Inlife innovation, the users, and the socio-political context. Inlife was evaluated as a useful instrument for efficient central care coordination and mutual involvement. This study emphasizes that the personal attitudes of the Inlife users to seek and provide support warrant attention, next to the characteristics of the actual Inlife innovation for optimal intervention uptake. Online and offline support might be integrated to raise awareness of caregiver social support needs and attitudes and provide insight into caregivers' available social capital. Dutch trial register NTR6131, Registered on 20 October 2016.


Preoperative chemoradiation and IOERT for unresectable or borderline resectable pancreas cancer.
Pre-operative chemoradiation (preop CRT) plus intraoperative electron irradiation (IOERT) has been used in the multidisciplinary treatment for patients with locally advanced unresectable or borderline resectable pancreas cancer. This review was performed to evaluate survival, relapse patterns and prognostic factors in patients treated with curative intent. Between January 2002 and December 2010, 48 patients with locally advanced pancreatic ductal adenocarcinoma received preop CRT prior to an attempt at resection and IOERT. 31/48 (65%) patients proceeded to curative-intent surgical resection. Resection status prior to preop CRT was locally unresectable (20 patients) and borderline resectable (11 patients). Preop CRT (45-50.4 Gy/25-28 Fx in 27/31) was delivered with concurrent 5FU or gemcitabine-based regimens. Subsequent gross total resection was achieved in 16 patients (R0, 11; R1, 5). IOERT was delivered in 28 patients (dose, 10-20 Gy). 16 patients also received adjuvant post-operative systemic chemotherapy. Outcomes evaluated include survival, local failure in the EBRT field (LF), central failure in the IOERT field (CF), and distant metastases. Resection status was predictive for survival and for patterns of relapse. For patients with at least a gross total resection after preop CRT (R0/R1; n=16) vs. no resection (n=15), both median and overall survival were improved (median 23 vs. 10 months; 2-year, 40% vs. 17%; 3-year, 40% vs. 0%; P=0.002). Liver or peritoneal relapse was documented in 22/31 patients (71%); LF/CF in 5/26 (16%). Long term survival and disease control are achievable in select patients with borderline resectable or locally unresectable pancreas cancer when gross total surgical resection is achieved after preop CRT. Continued evaluation of curative-intent combined modality therapy is warranted in this high risk population, but additional strategies are needed to improve resectability and disease control.


Antimicrobial Activity of Basil, Oregano, and Thyme Essential Oils.
For centuries, plants have been used for a wide variety of purposes, from treating infectious diseases to food preservation and perfume production. Presently, the increasing resistance of microorganisms to currently used antimicrobials in combination with the appearance of emerging diseases requires the urgent development of new, more effective drugs. Plants, due to the large biological and structural diversity of their components, constitute a unique and renewable source for the discovery of new antibacterial, antifungal, and antiparasitic compounds. In the present paper, the history, composition, and antimicrobial activities of the basil, oregano, and thyme essential oils are reviewed.


Characterization of proliferation and cellular wound fill in periodontal cells using an in vitro wound model.
The therapeutic success of periodontal regenerative therapy may be compromised by our limited understanding of the wound healing process. Wound healing requires the coordination of complex cellular and molecular interactions. Recently, using an in vitro wound model, our laboratory has shown that gingival fibroblasts (GF) fill an in vitro wound more rapidly than periodontal ligament cells (PDL). This suggests that there may be differences in the levels of proliferation for these 2 cell types during the wound healing process. Such specific cell type differences may be significant in clinical outcomes of regenerative therapy. Therefore, the aim of this research was to characterize and compare the levels of both proliferation and cellular wound fill between GF and PDL using our in vitro wound model. Primary cultures of human PDL and GF cells were established from explanted tissue, and passaged to 12-well tissue culture plates. Triplicate cultures of both cell types were grown to confluence and in vitro wounds were mechanically created, removing a 3 mm wide band of the cell layer across the diameter of the wells. The wells were then incubated for 2, 6, or 9 days in media containing either 0.1% or 10% fetal bovine serum (FBS). At each time point, cells were pulsed with 5-bromo, 2-deoxyuridine (BrdU), fixed, and nuclei stained to measure DNA synthesis (as a measure for proliferation). Cells were counter stained with cytoplasmic stain to measure cell number. Quantitative analysis distant from (area of interest [AOI 1]), next to (AOI 2), and within the wound boundaries (AOI 3 and 4) was accomplished using computer-assisted histomorphometry. The levels of proliferation and cellular fill for each cell type were assessed relative to time and AOI. Overall, the PDL displayed greater (P <0.01) levels of proliferation than the GF. For both cell types, proliferation was found to be significantly (P<0.001) greater at day 2 compared to other time points. PDL displayed greater levels of proliferation than GF in all AOI, with this difference reaching significance (P<0.02) within the cell layer (AOI 1 and 2). When comparing levels of cellular fill in 10% FBS, GF displayed greater wound fill than the PDL. This difference was significant at day 6 (P <0.05) for both the marginal (AOI 3) and central (AOI 4) portions of the wound. These findings, demonstrating unique differences between PDL and GF with respect to proliferation and wound fill in an in vitro model, suggest that there may be cell-specific differences in cellular activity critical to periodontal wound healing. In addition, the results of this study show that the cellular proliferation response may not accurately reflect the overall wound healing response.


Anterior cingulate Glutamate-Glutamine cycle metabolites are altered in euthymic bipolar I disorder.
Bipolar disorder (BD) has been consistently associated with abnormalities in the Glutamate/GABA-Glutamine cycle. Magnetic resonance spectroscopy (MRS) studies have reported increased brain Glutamate (Glu) and Glx (Glu+Glutamine) in subjects with BD. However, data on separate measures of GABA and Glutamine (Gln) in BD are sparse due to overlapping resonant signals. The development of new sequence methods in the quantification of these metabolites has allowed a better understanding of the Glu/GABA-Gln cycle but data on this field of research remains sparse in BD. Eighty-eight subjects (50 euthymic BD and 38 HC) underwent 3T proton magnetic resonance spectroscopy (1H MRS) in the anterior cingulate cortex (ACC; 2×2×4.5cm(3)) using a two-dimensional JPRESS sequence. GABA, Glutamine (Gln) and Glutamate (Glu) were quantified with the ProFit program. Using image segmentation and known creatine (Cre) concentrations for white and grey matter, metabolite concentrations were calculated for the excited MRS voxel. GABA levels did not differ between groups. Gln level was higher in euthymic BD patients than in healthy controls. The Glu level and Glu/Gln ratio were lower in BD patients than in controls. The use of anticonvulsants was associated with Gln increase but did not affect Glu or Glu/Gln. Neither lithium nor antipsychotic use influenced metabolite levels. The ACC MRS findings indicate that the glutamatergic function in euthymic medicated BD patients is altered relative to controls. Whether this feature is a metabolic signature of euthymic BD subjects should be the focus of future studies.


Phosphorylated tau in the retina correlates with tau pathology in the brain in Alzheimer's disease and primary tauopathies.
The retina is a potential source of biomarkers for the detection of neurodegenerative diseases. Accumulation of phosphorylated tau (p-tau) in the brain is a pathological feature characteristic for Alzheimer's disease (AD) and primary tauopathies. In this study the presence of p-tau in the retina in relation to tau pathology in the brain was assessed. Post-mortem eyes and brains were collected through the Netherlands Brain Bank from donors with AD (n = 17), primary tauopathies (n = 8), α-synucleinopathies (n = 13), other neurodegenerative diseases including non-tau frontotemporal lobar degeneration (FTLD) (n = 9), and controls (n = 15). Retina cross-sections were assessed by immunohistochemistry using antibodies directed against total tau (HT7), 3R and 4R tau isoforms (RD3, RD4), and phospho-epitopes Ser202/Thr205 (AT8), Thr217 (anti-T217), Thr212/Ser214 (AT100), Thr181 (AT270), Ser396 (anti-pS396) and Ser422 (anti-pS422). Retinal tau load was compared to p-tau Ser202/Thr205 and p-tau Thr217 load in various brain regions. Total tau, 3R and 4R tau isoforms were most prominently present in the inner plexiform layer (IPL) and outer plexiform layer (OPL) of the retina and were detected in all cases and controls as a diffuse and somatodendritic signal. Total tau, p-tau Ser202/Thr205 and p-tau Thr217 was observed in amacrine and horizontal cells of the inner nuclear layer (INL). Various antibodies directed against phospho-epitopes of tau showed immunoreactivity in the IPL, OPL, and INL. P-tau Ser202/Thr205 and Thr217 showed significant discrimination between AD and other tauopathies, and non-tauopathy cases including controls. Whilst immunopositivity was observed for p-tau Thr212/Ser214, Thr181 and Ser396, there were no group differences. P-tau Ser422 did not show any immunoreactivity in the retina. The presence of retinal p-tau Ser202/Thr205 and Thr217 correlated with Braak stage for NFTs and with the presence of p-tau Ser202/Thr205 in hippocampus and cortical brain regions. Depending on the phospho-epitope, p-tau in the retina is a potential biomarker for AD and primary tauopathies.


Platelet function in hairy-cell leukaemia.
A quantitative study of various aspects of platelet function was carried out in eight patients with typical hairy-cell leukaemia (HCL). In at least two patients platelet aggregation was convincingly reduced to more than one aggregating agent (ADP, adrenaline, collagen, thrombin, and ristocetin). Granular storage capacity for {(14)C} 5-HT was reduced in five of the six patients tested. The two patients with definitely abnormal aggregation had the greatest reduction in granular storage pool and the longest bleeding times of those tested but, like the other patients, they did not have a clinical haemostatic defect. It was concluded that a granular storage pool defect (SPD) was at least partly responsible for aggregation abnormalities in HCL since the platelet release reaction in response to thrombin appeared to be normal. All our patients ran a chronic course uncomplicated by any of the factors known to predispose to a platelet SPD acquired in the circulation. Although in the one patient tested before and after splenectomy there was some improvement in platelet aggregation after operation, there was no clear general relationship between defective platelet function and either previous splenectomy or platelet count. Since a direct involvement of the megakaryocytic series in the underlying cell proliferation of HCL seems unlikely, it is concluded that the platelet defect can most reasonably be attributed to the production of abnormal platelets as a result of marrow fibrosis and/or infiltration by hairy cells.


Leptin (rs7799039) and solute carrier family 30 zinc transporter (rs13266634) polymorphisms in Euro-Brazilian pregnant women with gestational diabetes.
Leptin (LEP), a protein that plays a fundamental role in the metabolism of energy reserves, and the solute carrier family 30 A8 zinc transporter (SLC30A8) have been consistently associated with diabetes. Women with gestational diabetes are at moderate risk of developing diabetes type 1 and 2 after pregnancy, in addition to complications to the fetus. We investigated the association of the polymorphisms rs7799039 (LEP) and rs13266634 (SLC30A8) in a case-control study in Euro-Brazilians with gestational diabetes (GDM, N = 134) and healthy pregnant women (control, N = 180). Real-time PCR with fluorescent probes (TaqMan system) was applied to genotyping. All polymorphisms were in Hardy-Weinberg equilibrium. The minor allele frequencies, for healthy and GDM, respectively, for the A-allele (LEP gene rs7799039) were 40.3% (95%CI = 35-45%) vs 36.6% (95%CI = 31-42%), P = 0.345; and for the T-allele (SLC30A8 gene rs13266634) were 27.8% (95%CI = 23-32%) vs 23.5% (95%CI = 18-29%), P = 0.227. Genotype comparisons for both polymorphisms showed no significant difference (P > 0.05). The polymorphisms rs7799039 and rs13266634 were not associated with GDM in the population studied (P > 0.05). The minor allele frequencies for both polymorphisms were similar to those of other Caucasian populations.


Bacterial autolysins trim cell surface peptidoglycan to prevent detection by the Drosophila innate immune system.
Bacteria have to avoid recognition by the host immune system in order to establish a successful infection. Peptidoglycan, the principal constituent of virtually all bacterial surfaces, is a specific molecular signature recognized by dedicated host receptors, present in animals and plants, which trigger an immune response. Here we report that autolysins from Gram-positive pathogenic bacteria, enzymes capable of hydrolyzing peptidoglycan, have a major role in concealing this inflammatory molecule from Drosophila peptidoglycan recognition proteins (PGRPs). We show that autolysins trim the outermost peptidoglycan fragments and that in their absence bacterial virulence is impaired, as PGRPs can directly recognize leftover peptidoglycan extending beyond the external layers of bacterial proteins and polysaccharides. The activity of autolysins is not restricted to the producer cells but can also alter the surface of neighboring bacteria, facilitating the survival of the entire population in the infected host. DOI: http://dx.doi.org/10.7554/eLife.02277.001.


Management of perforating colon trauma: randomization between primary closure and exteriorization.
During a 44 month trial, 268 patients with wounds of the colon were entered into a prospective, randomized, nonblinded study. Consideration for primary closure demanded that: preoperative shock was never profound, blood loss was less than 20% of estimated normal volume, no more than two intra-abdominal organ systems had been injured, fecal contamination was minimal, operation was begun within eight hours, and wounds of colon and abdominal wall were never so destructive as to require resection. Once such criteria had been satisfied, colon wound management was dictated by last digit in the randomly assigned hospital number; odd indicated primary closure; even, exteriorization of the wound or primary closure with protection by a proximal vent. Results obtained in 139 determinant patients eligible for randomization revealed that primary closure (67 patients) had a lower infection rate of the incision (48% vs S7%, p > 0.05) and a still lower infection rate for the abdomen proper (15% vs 29%, p < 0.05) on comparison to the 72 patients with a randomized colostomy. Morbidity otherwise for the randomized colostomy was tenfold greater than if a primary closure had been performed. Average postoperative stay was six days longer (p < 0.01) if a colostomy had been created, exclusive of subsequent hospitalization for colostomy closure; while the total extra cost for management of the colon wound by colostomy was approximately $2,700.00. Although immediate mortalities were identical, one late death occurred following colostomy closure. These data not only confirm the safety of primary closure for colon wounds in selected cases, but also indicate that such should become the preferred method of treatment whenever specific criteria have been met.


A single regulatory gene is sufficient to alter Vibrio aestuarianus pathogenicity in oysters.
Oyster diseases caused by pathogenic vibrios pose a major challenge to the sustainability of oyster farming. In France, since 2012 a disease affecting specifically adult oysters has been associated with the presence of Vibrio aestuarianus. Here, by combining genome comparison, phylogenetic analyses and high-throughput infections of strains isolated before or during the recent outbreaks, we show that virulent strains cluster into two V. aestuarianus lineages independently of the sampling dates. The bacterial lethal dose was not different between strains isolated before or after 2012. Hence, the emergence of a new highly virulent clonal strain is unlikely. Each lineage comprises nearly identical strains, the majority of them being virulent, suggesting that within these phylogenetically coherent virulent lineages a few strains have lost their pathogenicity. Comparative genomics allowed the identification of a single frameshift in a non-virulent strain. This mutation affects the varS gene that codes for a signal transduction histidine-protein kinase. Genetic analyses confirmed that varS is necessary for infection of oysters and for a secreted metalloprotease expression. For the first time in a Vibrio species, we show here that VarS is a key factor of pathogenicity.


miR-138-Mediated Regulation of KINDLIN-2 Expression Modulates Sensitivity to Chemotherapeutics.
Prostate cancer is the second leading cause of cancer-related death in men, second only to lung cancer, mainly due to disease reoccurrence as a result to lack of response to androgen deprivation therapies (ADT) after castration. Patients with metastatic castration-resistant prostate cancer (mCRPC) have very limited treatment options, with docetaxel as the first-line standard of care, for which resistance to this chemotherapeutic ultimately develops. Therefore, finding ways to sensitize tumors to chemotherapies and to limit chemoresistance provides a viable strategy to extend the survival of mCRPC patients. This study investigated the role of Kindlin-2 (FERMT2/K2), a member of the Kindlin family of FERM domain proteins and key regulators of the adhesive functions mediated by integrin, in the sensitization of mCRPC to chemotherapeutics. Loss of K2, which is overexpressed in prostate cancer cells derived from mCRPC tumors, compared with those cells derived from androgen-dependent tumors, significantly enhanced apoptosis and cell death of docetaxel-treated PC3 cells. Furthermore, it was determined that K2-mediated sensitization to docetaxel treatment is the result of inhibition of β1-integrin signaling. Finally, miR-138 specifically targeted K2 and inhibited its expression, thereby regulating a miR-138/K2/β1-integrin signaling axis in mCRPC that is critical for the modulation of sensitivity to chemotherapeutics. Thus, these data identify a novel signaling axis where K2 in combination with chemotherapeutics provides a new target for the treatment of mCRPC. Targeted inhibition of Kindlin-2 in combination with chemotherapy represents an effective treatment option for mCRPC.


Multiscale modeling and decoding algorithms for spike-field activity.
Behavior is encoded across multiple spatiotemporal scales of brain activity. Modern technology can simultaneously record various scales, from spiking of individual neurons to large neural populations measured with field activity. This capability necessitates developing multiscale modeling and decoding algorithms for spike-field activity, which is challenging because of the fundamental differences in statistical characteristics and time-scales of these signals. Spikes are binary-valued with a millisecond time-scale while fields are continuous-valued with slower time-scales. We develop a multiscale encoding model, adaptive learning algorithm, and decoder that explicitly incorporate the different statistical profiles and time-scales of spikes and fields. The multiscale model consists of combined point process and Gaussian process likelihood functions. The multiscale filter (MSF) for decoding runs at the millisecond time-scale of spikes while adding information from fields at their slower time-scales. The adaptive algorithm learns all spike-field multiscale model parameters simultaneously, in real time, and at their different time-scales. We validated the multiscale framework within motor tasks using both closed-loop brain-machine interface (BMI) simulations and non-human primate (NHP) spike and local field potential (LFP) motor cortical activity during a naturalistic 3D reach task. Our closed-loop simulations show that the MSF can add information across scales and that the adaptive MSF can accurately learn all parameters in real time. We also decoded the seven joint angular trajectories of the NHP arm using spike-LFP activity. These data showed that the MSF outperformed single-scale decoding, this improvement was due to the addition of information across scales rather than the dominance of one scale and was largest in the low-information regime, and the improvement was similar regardless of the degree of overlap between spike and LFP channels. This multiscale framework provides a tool to study encoding across scales and may help enhance future neurotechnologies such as motor BMIs.


Pharmacokinetics, toxicity, and functional studies of the selective Kv1.3 channel blocker 5-(4-phenoxybutoxy)psoralen in rhesus macaques.
The small molecule 5-(4-phenoxybutoxy)psoralen (PAP-1) is a selective blocker of the voltage-gated potassium channel Kv1.3 that is highly expressed in cell membranes of activated effector memory T cells (TEMs). The blockade of Kv1.3 results in membrane depolarization and inhibition of TEM proliferation and function. In this study, the in vitro effects of PAP-1 on T cells and the in vivo toxicity and pharmacokinetics (PK) were examined in rhesus macaques (RM) with the ultimate aim of utilizing PAP-1 to define the role of TEMs in RM infected with simian immunodeficiency virus (SIV). Electrophysiologic studies on T cells in RM revealed a Kv1.3 expression pattern similar to that in human T cells. Thus, PAP-1 effectively suppressed TEM proliferation in RM. When administered intravenously, PAP-1 showed a half-life of 6.4 hrs; the volume of distribution suggested extensive distribution into extravascular compartments. When orally administered, PAP-1 was efficiently absorbed. Plasma concentrations in RM undergoing a 30-day, chronic dosing study indicated that PAP-1 levels suppressive to TEMs in vitro can be achieved and maintained in vivo at a non-toxic dose. PAP-1 selectively inhibited the TEM function in vivo, as indicated by a modest reactivation of cytomegalovirus (CMV) replication. Immunization of these chronically treated RM with the live influenza A/PR8 (flu) virus suggested that the development of an in vivo, flu-specific, central memory response was unaffected by PAP-1. These RM remained disease-free during the entire course of the PAP-1 study. Collectively, these data provide a rational basis for future studies with PAP-1 in SIV-infected RM.


Comparative study of the effects of phosphatidylcholine rich in DHA and EPA on Alzheimer's disease and the possible mechanisms in CHO-APP/PS1 cells and SAMP8 mice.
Metabolic stress induced by a high-fat (HF) diet leads to cognitive dysfunction and aging. In the present study, Chinese hamster ovary cells stably transfected with amyloid precursor protein (APP) and presenilin 1 (PS1) (CHO-APP/PS1 cells) and SAMP8 mice fed with an HF diet were used to study the effects of docosahexaenoic acid (DHA)-enriched phosphatidylcholine (DHA-PC) and eicosapentaenoic acid (EPA)-enriched phosphatidylcholine (EPA-PC) on Alzheimer's disease (AD) and the possible mechanisms involved in these effects. Behavior test results indicated that DHA-PC exerted better effects than EPA-PC on improving memory and cognitive deficiency. Further analysis showed that DHA-PC and EPA-PC could significantly decrease β-amyloid (Aβ) concentrations in CHO-APP/PS1 cells and SAMP8 mice by inhibiting APP, PS1, and BACE1 expression. Moreover, both DHA-PC and EPA-PC can increase the activities of the antioxidant index, including SOD, T-AOC, GSH, and GSH-PX, and inhibit levels of MDA, NO, and NOS. In addition, the expressions of inflammatory factors (TNF-α, IL-1β) and apoptosis were significantly suppressed via improving the ratio of Bcl-2/Bax and decreasing the expression of pro-apoptosis factors. Interestingly, only DHA-PC could improve the expression of neurotrophic factors, including BDNF, synaptophysin, and growth associated protein 43. DHA-PC and EPA-PC could ameliorate memory and cognitive function of HF diet-fed SAMP8 mice via inhibiting Aβ generation, suppressing oxidative stress and apoptosis, down-regulating inflammatory response, and improving neurotrophic activity. Therefore, DHA-PC and EPA-PC may be applied as food supplements and/or functional ingredients to relieve neurodegenerative disease.


Inversion effects in face-selective cortex with combinations of face parts.
The face inversion effect has been used as a basis for claims about the specialization of face-related perceptual and neural processes. One of these claims is that the fusiform face area (FFA) is the site of face-specific feature-based and/or configural/holistic processes that are responsible for producing the face inversion effect. However, the studies on which these claims were based almost exclusively used stimulus manipulations of whole faces. Here, we tested inversion effects using single, discrete features and combinations of multiple discrete features, in addition to whole faces, using both behavioral and fMRI measurements. In agreement with previous studies, we found behavioral inversion effects with whole faces and no inversion effects with a single eye stimulus or the two eyes in combination. However, we also found behavioral inversion effects with feature combination stimuli that included features in the top and bottom halves (eyes-mouth and eyes-nose-mouth). Activation in the FFA showed an inversion effect for the whole-face stimulus only, which did not match the behavioral pattern. Instead, a pattern of activation consistent with the behavior was found in the bilateral inferior frontal gyrus, which is a component of the extended face-preferring network. The results appear inconsistent with claims that the FFA is the site of face-specific feature-based and/or configural/holistic processes that are responsible for producing the face inversion effect. They are more consistent with claims that the FFA shows a stimulus preference for whole upright faces.


Mapping whispering-gallery modes in microspheres with a near-field probe.
We demonstrate the use of a near-field probe to map the angular dependence of high-Q whispering-gallery modes in fused-silica microspheres. The mapping is performed by placing a micrometer-sized tip formed on the end of a monomode fiber into the evanescent f ield at the microsphere surface, causing light to be coupled from the microsphere resonance into the fiber guided mode. The light output of the fiber is then measured while the tip is moved to different points on the microsphere surface. We have used this method to investigate the lifting of spherical degeneracy in the system.


Non-Newtonian Polymer-Nanoparticle Hydrogels Enhance Cell Viability during Injection.
Drug delivery and cell transplantation require minimally invasive deployment strategies such as injection through clinically relevant high-gauge needles. Supramolecular hydrogels comprising dodecyl-modified hydroxypropylmethylcellulose and poly(ethylene glycol)-block-poly(lactic acid) have been previously demonstrated for the delivery of drugs and proteins. Here, it is demonstrated that the rheological properties of these hydrogels allow for facile injectability, an increase of cell viability after injection when compared to cell viabilities of cells injected in phosphate-buffered saline, and homogeneous cell suspensions that do not settle. These hydrogels are injected at 1 mL min-1 with pressures less than 400 kPa, despite the solid-like properties of the gel when at rest. The cell viabilities immediately after injection are greater than 86% for adult human dermal fibroblasts, human umbilical vein cells, smooth muscle cells, and human mesenchymal stem cells. Cells are shown to remain suspended and proliferate in the hydrogel at the same rate as observed in cell media. The work expands on the versatility of these hydrogels and lays a foundation for the codelivery of drugs, proteins, and cells.


The consequences of monoecy and protogyny for mating in wind-pollinated Carex.
Monoecy and protogyny are widespread in wind-pollinated plants and have been interpreted as outcrossing mechanisms, though few studies have investigated their function. Carex, a large genus of anemophilous herbs, is predominantly monoecious and many species are protogynous. We investigated whether monoecy and protogyny limit self-pollination in seven Carex species. We conducted field experiments comparing stigmatic pollen loads and seed set between intact and emasculated stems. We tested for self-compatibility and evaluated pollen limitation of seed set by supplemental pollination. Finally, we measured outcrossing rates in open-pollinated and emasculated stems using allozyme markers. Emasculated stems captured significantly less pollen than open-pollinated stems and set less seed. Pollen deposition during the female-only phase for intact stems was only 12% of the total captured. Outcrossing rates for three species indicated high selfing (range t = 0.03-0.39). Allozyme loci in the remaining species were monomorphic also suggesting high selfing. These results demonstrate that neither monoecy nor protogyny is particularly effective at limiting self-fertilization. Selection for the avoidance of selfing is unlikely to maintain monoecy in many Carex species although protogyny may provide limited opportunities for outcrossing. We propose that geitonogamy in self-compatible wind-pollinated species with unisexual flowers may be widespread and provides reproductive assurance.


Two-photon imaging of spinal cord cellular networks.
Two-photon microscopy enables high-resolution in vivo imaging of cellular morphology and activity, in particular of population activity in complex neuronal circuits. While two-photon imaging has been extensively used in a variety of brain regions in different species, in vivo application to the vertebrate spinal cord has lagged behind and only recently became feasible by adapting and refining the experimental preparations. A major experimental challenge for spinal cord imaging is adequate control of tissue movement, which meanwhile can be achieved by various means. One set of studies monitored structural dynamics of neuronal and glial cellular components in living animals using transgenic mice with specific expression of fluorescent proteins. Other studies employed in vivo calcium imaging for functional measurements of sensory-evoked responses in individual neurons of the dorsal horn circuitry, which at present is the only part of rodent spinal cord grey matter accessible for in vivo imaging. In a parallel approach, several research groups have applied two-photon imaging to sensorimotor circuits in the isolated spinal cord (in vitro) to provide complementary information and valuable new perspectives on the function of specific interneuron types in locomotor-related networks. In this review we summarize recent results from these types of high-resolution two-photon imaging studies in the spinal cord and provide experimental perspectives for improving and extending this approach in future applications.


Hypertonic dextrose injections (prolotherapy) for knee osteoarthritis: results of a single-arm uncontrolled study with 1-year follow-up.
The objective of this study was to determine whether prolotherapy, an injection-based complementary treatment for chronic musculoskeletal conditions, improves pain, stiffness, and function in adults with symptomatic knee osteoarthritis (KOA) compared to baseline status. This was a prospective, uncontrolled study with 1-year follow-up. The study was conducted in an outpatient setting. Adults with at least 3 months of symptomatic KOA, recruited from clinical and community settings, participated in the study. Participants received extra-articular injections of 15% dextrose and intra-articular prolotherapy injections of 25% dextrose at 1, 5, and 9 weeks, with as-needed treatments at weeks 13 and 17. Primary outcome measure was the validated Western Ontario McMaster University Osteoarthritis Index (WOMAC). Secondary outcome measure was the validated Knee Pain Scale (KPS). Tertiary outcome measure was procedure-related pain severity and participant satisfaction. Thirty-six (36) participants (60 ± 8.7 years old, 21 female) with moderate-to-severe KOA received an average of 4.3 ± 0.7 prolotherapy injection sessions over a 17-week treatment period and reported progressively improved scores during the 52-week study on WOMAC and KPS measures. Participants reported overall WOMAC score improvement 4 weeks after the first injection session (7.6 ± 2.4 points, 17.2%), and continued to improve through the 52-week follow-up (15.9 ± 2.5 points, p<0.001, 36.1%). KPS scores improved in both injected (p<0.001) and uninjected knees (p<0.05). Prescribed low-dose opioid analgesia effectively treated procedure-related pain. Satisfaction was high and there were no adverse events. Female gender, age 46-65 years old, and body-mass index of 25 kg/m(2) or less were associated with greater improvement on the WOMAC instrument. In adults with moderate to severe KOA, dextrose prolotherapy may result in safe, significant, sustained improvement of knee pain, function, and stiffness scores. Randomized multidisciplinary effectiveness trials including evaluation of potential disease modification are warranted to further assess the effects of prolotherapy for KOA.


Cardiorespiratory Fitness Is Associated with Selective Attention in Healthy Male High-School Students.
Background: Previous studies have shown associations of physical fitness and cognition in children and in younger and older adults. However, knowledge about associations in high-school adolescents and young adults is sparse. Thus, the aim of this study was to evaluate the association of physical fitness, measured as maximal oxygen uptake ([Formula: see text]), muscle mass, weekly training, and cognitive function in the executive domains of selective attention and inhibitory control, in healthy male high-school students. Methods: Fifty-four males (17.9 ± 0.9 years, 72 ± 11 kg and 182 ± 7 cm) completed a [Formula: see text] test, a body composition test and a visual cognitive task based on the Posner cue paradigm with three types of stimuli with different attentional demands (i.e., stimuli presentation following no cue, valid cue or invalid cue presentations). The task consisted of 336 target stimuli, where 56 (17%) of the target stimuli appeared without a cue (no cue), 224 (67%) appeared in the same rectangle as the cue (valid cue) and 56 (17%) appeared in the rectangle opposite to the cue (invalid cue). Mean reaction time (RT) and corresponding errors was calculated for each stimuli type. Total task duration was 9 min and 20 s In addition, relevant background information was obtained in a questionnaire. Results: Linear mixed model analyses showed that higher [Formula: see text] was associated with faster RT for stimuli following invalid cue (Estimate = -2.69, SE = 1.03, p = 0.011), and for stimuli following valid cue (Estimate = -2.08, SE = 1.03, p = 0.048). There was no association of muscle mass and stimuli (F = 1.01, p = 0.397) or of weekly training and stimuli (F = 0.99, p = 0.405). Conclusion: The results suggest that cardiorespiratory fitness is associated with cognitive performance in healthy male high-school students in the executive domains of selective attention.


Acute respiratory distress syndrome.
Acute respiratory distress syndrome is a common cause of acute respiratory failure that is underdiagnosed both inside and outside of intensive care units. Progression to the most severe forms of the syndrome confers a mortality rate greater than 40% and is associated with often severe functional disability and psychological sequelae in survivors. While there are no disease-modifying pharmacotherapies for the syndrome, this progression may be prevented through the institution of quality improvement measures that minimise iatrogenic injury associated with acute severe illness.