Modelling of blood pressure and total cardiovascular risk outcomes after second-line valsartan therapy: the BSCORE study.
European guidelines recommend that antihypertensive management should be graded as a function of total cardiovascular risk. To examine the multilevel (patient- and physician-level) determinants of blood pressure and residual total cardiovascular risk outcomes associated with second-line valsartan therapy. The BSCORE study was a prospective, multi-centre, pharmacoepidemiological study of the "real-world" effectiveness of second-line valsartan with or without hydrochlorothiazide. A total of 3497 patients were recruited by 354 physicians. Mean age was 63.8±12.0 years; 52.3% were male; 20.9% were smokers; 47.7% were dyslipidaemic; and 23.6% had diabetes. On average, reductions in blood pressure and increases in the proportions of patients with controlled blood pressure after 90 days were statistically significant (all P<0.001). Twenty-one percent of systolic blood pressure and 25.6% of diastolic blood pressure variability at follow-up was attributable to physician-level characteristics. Significant reductions in total cardiovascular risk were observed (P<0.001); with 12.5% of the variability in total cardiovascular risk change attributable to physician-level characteristics. Several independent determinants of blood pressure outcomes were identified, many of which are modifiable. Second-line valsartan therapy improves blood pressure outcomes under variable real-world conditions, and is associated with a decrease in total cardiovascular risk. Optimizing antihypertensive effectiveness, including the reduction of residual cardiovascular risk, involves managing concomitant conditions and risk factors, improving adherence, and identifying physician-level factors amenable to intervention.


A distance learning model in a physical therapy curriculum.
In response to the rural health initiative established in 1991, the University of Kentucky has developed an innovative distance learning program of physical therapy instruction that combines classroom lecture and discussion via compressed video technology with laboratory experiences. The authors describe the process of planning, implementing, and evaluating a specific distance learning course in pathomechanics for the professional-level master's-degree physical therapy students at the University of Kentucky. This presentation may serve as a model for teaching distance learning. Descriptions of optimal approaches to preclass preparation, scheduling, course delivery, use of audiovisual aids, use of handout material, and video production are given. Special activities that may enhance or deter the achievement of the learning objectives are outlined, and a problem-solving approach to common problems encountered is presented. An approach to evaluating and comparing course outcomes for the distance learnere is presented. For this particular course, there was no statistically significant difference in the outcome measures utilized to compare the distance learners with the on-site learners.


Regeneration and outcome of dual grafts in living donor liver transplantation.
In living donor liver transplantation (LDLT), the essential aims are to provide an adequate graft volume to the recipient and to keep a sufficient remnant liver volume in the donor. In some instances, these aims cannot be met by a single donor and LDLT using dual grafts from two donors is a good solution. From 2002 to 2009, five recipients in our hospital received dual graft LDLT. Two recipients received one right lobe and one left lobe grafts; the other three received two left lobe grafts. The mean final liver regeneration rate was 91.2%. Left lobe graft atrophy in the long term was observed in recipients who received a right and a left lobe grafts. The initial bigger volume graft in all recipients was noted to have better regeneration than the smaller volume grafts. Portal flow and bilateral grafts volume size discrepancy were considered as two major factors influencing graft regeneration in this study. We also noted that the initial graft volume correlated with portal flow in the separate grafts and finally contribute to individual graft regeneration. Because of compensatory hypertrophy of the other graft, recipients who experienced atrophy of one graft did not show signs of liver dysfunction.


Glutaric acidemia, type I, missed by newborn screening in an infant with dystonia following promethazine administration.
We report a child initially diagnosed with promethazine-induced dystonia despite a lack of response to diphenhydramine therapy. On further evaluation, the child was diagnosed with glutaric acidemia, type I (GA-I), an autosomal recessive inborn error of metabolism caused by the deficiency of glutaryl-CoA dehydrogenase. The characteristic clinical feature of GA-I is an acute encephalopathic and neurologic crisis typically occurring during a catabolic state. Despite slow improvement, many patients do not fully recover from a neurologic crisis, and residual neurologic morbidity can be significant. Although newborn screening using tandem mass spectrometry is expected to enable presymptomatic diagnosis of GA-I, this patient was not detected by newborn screening with tandem mass spectrometry. Therefore, a high suspicion of GA-I must be maintained in the evaluation of childhood dystonia, even when newborn screening results are reportedly normal.


Carbon nanotube/gold nanoparticle composite-coated membrane as a facile plasmon-enhanced interface for sensitive SERS sensing.
The facile assembly of three-dimensional (3D) plasmonic substrates has been demonstrated. The assembly is based on the homogeneous decoration of multi-walled carbon nanotube/gold nanoparticle (CNT/AuNP) hybrid nanocomposites on a commercial polyvinylidene difluoride (PVDF) membrane, which is achieved via simple filtration. The CNT/AuNP hybrids with a unique 1D/0D structure remarkably improve the coverage and uniformity of plasmonic nanostructures on the membrane. The effective inter-particle and inter-tube coupling creates a multitude of hot spots within the probe area, and can produce a strong SERS effect. Moreover, the flexible membrane-based scaffold can efficiently collect and concentrate trace targets from large-volume sample solutions at milliliter-scale. The membrane-based SERS sensor shows high sensitivity and good reproducibility. The SERS sensor is employed to detect various molecular contaminants in aqueous samples, demonstrating its excellent field-testing capabilities for applications ranging from food safety to environmental monitoring.


An experimental test of the Community Assembly by Trait Selection (CATS) model.
The Community Assembly by Trait Selection (CATS) model of community assembly predicts species abundances along environmental gradients in relatively undisturbed vegetation. Here we ask whether this model, when calibrated with data from natural plant communities, can predict the abundances of five dominant grass species (Bouteloua gracilis, Elymus elymoides, Festuca arizonica, Muhlenbergia montana, and Poa fendleriana) in a greenhouse experiment that manipulated light and soil properties. To address this question, we used generalized additive models (GAMs) to model community-weighted mean (CWM) seed mass, mean Julian flowering date, and specific root length (SRL) as non-linear functions of two environmental variables (soil pH and pine basal area) in natural vegetation. The model-fitted CWM traits were then used as constraints in the CATS model to predict the relative abundance of the five grass species that were seeded in a mixture at equal densities into a 2×2 factorial experiment with soil parent material and light level as crossed factors. Light was the most important factor influencing seedling community composition, especially the abundances of Bouteloua gracilis and Poa fendleriana. The model-predicted relative abundances were significantly correlated with the observed relative abundances, and the model accurately predicted the dominant species in every treatment. P. fendleriana was correctly predicted to be the most abundant species in both shade treatments and the sun-basalt treatment, and B. gracilis was correctly predicted to be the most abundant species in the sun-limestone treatment. Our results provide experimental evidence that environmental filtering of the species pool occurs in the early stages of community assembly (including germination, emergence, and early growth), and that trait-based models calibrated with data from natural plant communities can be used to predict the outcome of the early stages of community assembly under experimental conditions.


Kinetic mechanism of DNA polymerase I (Klenow fragment): identification of a second conformational change and evaluation of the internal equilibrium constant.
In a previously determined minimal kinetic scheme for DNA polymerization catalyzed by the Klenow fragment (KF) of Escherichia coli DNA polymerase I, a nonchemical step that interconverted the KF'.DNAn+1.PPi and KF.DNAn+1PPi complexes was not observed in correct incorporation [Kuchta, R. D., Mizrahi, V., Benkovic, P.A., Johnson, K.A., & Benkovic, S.J. (1987) Biochemistry 26, 8410-8417] but was detected in misincorporation [Kuchta, R. D., Benkovic, P.A., & Benkovic, S.J. (1988) Biochemistry 27, 6716-6725]. In a pulse-chase experiment in this study, a burst amplitude of 100% of the enzyme concentration is observed; under pulse-quench conditions, the burst amplitude is 80%, indicative of the accumulation of the KF'.DNA.dNTP species owing to a slow step subsequent to chemical bond formation. This latter step was unequivocally identified by single-turnover pyrophosphorolysis and pyrophosphate-exchange experiments as one interconverting KF'.DNAn+1.PPi and KF.DNAn+1.PPi. The rate constants for this step in both directions were established through the rate constants for processive synthesis and pyrophosphorolysis. Pyrophosphorolysis of a 3'-phosphorothioate DNA duplex confirmed that the large elemental effect observed previously [Mizrahi, V., Henrie, R. N., Marlier, J.F., Johnson, K.A., & Benkovic, S.J. (1985) Biochemistry 24, 4010-4018] in this direction but not in polymerization is due to a marked decrease in the affinity of KF for the phosphorothioate-substituted duplex and not to the chemical step. The combination of the experimentally measured equilibrium constant for the bound KF.DNA species with the collective kinetic measurements further extends previous insights into the dynamics of the polymerization process catalyzed by KF.


Association of dietary cholesterol and egg intakes with the risk of incident dementia or Alzheimer disease: the Kuopio Ischaemic Heart Disease Risk Factor Study.
There is little information about the associations of intakes of cholesterol and eggs, a major source of dietary cholesterol, with the risk of cognitive decline in general populations or in carriers of apolipoprotein E ɛ4 (APO-E4), a major risk factor for dementia. We investigated the associations of cholesterol and egg intakes with incident dementia, Alzheimer disease (AD), and cognitive performance in middle-aged and older men from Eastern Finland. A total of 2497 dementia-free men, aged 42-60 y in 1984-1989 at the baseline examinations of the prospective, population-based Kuopio Ischaemic Heart Disease Risk Factor Study, were included in the study. Information on the apolipoprotein E (Apo-E) phenotype was available for 1259 men. Data on cognitive performance tests at the 4-y re-examinations were available for 480 men. Dietary intakes were assessed with the use of 4-d food records at baseline. Dementia and AD diagnoses were based on Finnish health registers. Cox regression and ANCOVA were used for the analyses. During the 21.9-y follow-up, 337 men were diagnosed with dementia, and 266 men were diagnosed with AD. Neither cholesterol nor egg intake was associated with a higher risk of incident dementia or AD. For example, when evaluated continuously, each intake of 100 mg cholesterol/d was associated with a multivariable-adjusted HR of 0.90 (95% CI: 0.79, 1.02) for incident dementia, and each additional 0.5 egg (27 g)/d was associated with an HR of 0.89 (95% CI: 0.78, 1.01). However, egg intake was associated with better performance on neuropsychological tests of the frontal lobe and executive functioning, the Trail Making Test, and the Verbal Fluency Test. The Apo-E4 phenotype did not modify the associations of cholesterol or egg intake (P-interactions > 0.11). Neither cholesterol nor egg intake is associated with an increased risk of incident dementia or AD in Eastern Finnish men. Instead, moderate egg intake may have a beneficial association with certain areas of cognitive performance.


Oxidative stress reversibly inactivates myocardial enzymes during cardiac arrest.
Oxidative stress during cardiac arrest may inactivate myocardial enzymes and thereby exacerbate ischemic derangements of myocardial metabolism. This study examined the impact of cardiac arrest on left ventricular enzymes. Beagles were subjected to 5 min of cardiac arrest and 5 min of open-chest cardiac compressions (OCCC) before epicardial direct current countershocks were applied to restore sinus rhythm. Glutathione/glutathione disulfide redox state (GSH/GSSG) and a panel of enzyme activities were measured in snap-frozen left ventricle. To test whether oxidative stress during arrest inactivated the enzymes, metabolic (pyruvate) or pharmacological (N-acetyl-l-cysteine) antioxidants were infused intravenously for 30 min before arrest. During cardiac arrest, activities of phosphofructokinase, citrate synthase, aconitase, malate dehydrogenase, creatine kinase, glucose-6-phosphate dehydrogenase, and glutathione reductase fell by 56, 81, 55, 34, 42, 55, and 45%, respectively, coincident with 50% decline in GSH/GSSG. OCCC effected full recovery of glutathione reductase and partial recovery of citrate synthase and aconitase, in parallel with GSH/GSSG. Phosphofructokinase, malate dehydrogenase, creatine kinase, and glucose-6-phosphate dehydrogenase recovered only after cardioversion. Antioxidant pretreatments augmented phosphofructokinase, aconitase, and malate dehydrogenase activities before arrest and enhanced these activities, as well as those of citrate synthase and glucose-6-phosphate dehydrogenase, during arrest. In conclusion, cardiac arrest reversibly inactivates several important myocardial metabolic enzymes. Antioxidant protection of these enzymes implicates oxidative stress as a principal mechanism of enzyme inactivation during arrest.


Rhodococcus gordoniae sp. nov., an actinomycete isolated from clinical material and phenol-contaminated soil.
The taxonomic relationships of two actinomycetes provisionally assigned to the genus Rhodococcus were determined using a polyphasic taxonomic approach. The generic assignment was confirmed by 16S rRNA gene similarity data, as the organisms, strains MTCC 1534 and W 4937(T), were shown to belong to the Rhodococcus rhodochrous subclade. These organisms had phenotypic properties typical of rhodococci; they were aerobic, Gram-positive, weakly acid-fast actinomycetes that showed an elementary branching-rod-coccus growth cycle and contained meso-diaminopimelic acid, arabinose and galactose in whole-organism hydrolysates, N-glycolated muramic acid residues, dehydrogenated menaquinones with eight isoprene units as the predominant isoprenologue and mycolic acids that co-migrated with those extracted from the type strain of R. rhodochrous. The strains had identical phenotypic profiles and belong to the same genomic species, albeit one distinguished from Rhodococcus pyridinivorans, with which they formed a distinct phyletic line. They were also distinguished from representatives of all of the species classified in the R. rhodochrous 16S rRNA gene tree using a set of phenotypic features. The genotypic and phenotypic data show that the strains merit recognition as a novel species of Rhodococcus. The name proposed is Rhodococcus gordoniae sp. nov., with the type strain W 4937(T) (=DSM 44689(T)=NCTC 13296(T)).


Investigating and modelling pituitary endocrine network function.
Endocrine cells in the mammalian pituitary are arranged into three-dimensional homotypic networks that wire the gland and act to optimise hormone output by allowing the transmission of information between cell ensembles in a temporally precise manner. Despite this, the structure-function relationships that allow cells belonging to these networks to display coordinated activity remain relatively uncharacterised. This review discusses the recent technological advances that have allowed endocrine cell network structure and function to be probed and the mathematical models that can be used to analyse and present the resulting data. In particular, we focus on the mechanisms that allow endocrine cells to dynamically function as a population to drive hormone release as well as the experimental and theoretical methods that are used to track and model information flow through the network.


Thyroid autoimmunity and function among Ugandan children and adolescents with type-1 diabetes mellitus.
Up to 30% of type-1 diabetes mellitus (T1DM) patients have co-existent thyroid autoimmunity with up to 50% of them having associated thyroid dysfunction. Routine screening for thyroid autoimmunity and dysfunction is recommended in all T1DM patients. However, this was not currently practiced in Ugandan paediatric diabetes clinics. There was also paucity of data regarding thyroid autoimmunity and dysfunction in African children and adolescents with diabetes mellitus. The objective of this study was to quantify the magnitude of thyroid autoimmunity and dysfunction in Ugandan children with TIDM. This was a cross sectional descriptive study to determine the prevalence of thyroid autoantibodies and describe thyroid function among children and adolescents aged 1-19 years with diabetes mellitus attending the paediatric diabetes clinic at Mulago National Referral Hospital, Kampala, Uganda. Following enrollment, we obtained details of clinical history and performed physical examination. Blood (plasma) was assayed to determine levels of antibodies to thyroid peroxidase (antiTPO), free thyroxine (FT4) and thyrotropin (TSH). The prevalence of thyroid autoimmunity was 7.3% (5/69). All antiTPO positive subjects were post pubertal, aged between 13-17 years with females comprising 3/5 of the antiTPO positive subjects. All study subjects were clinically euthyroid; however, 7.3% (5/69) of the study subjects had subclinical hypothyroidism. These data strengthen the argument for routine screening of all diabetic children and adolescents for thyroid autoimmunity (particularly anti-TPO) as recommended by international guidelines. We also recommend evaluation of thyroid function in diabetic children and adolescents to minimize the risk of undiagnosed thyroid dysfunction.


Improving the teaching skills of residents as tutors/facilitators and addressing the shortage of faculty facilitators for PBL modules.
Residents play an important role in teaching of medical undergraduate students. Despite their importance in teaching undergraduates they are not involved in any formal training in teaching and leadership skills. We aimed to compare the teaching skills of residents with faculty in facilitating small group Problem Based Learning (PBL) sessions. This quasi experimental descriptive comparative research involved 5 postgraduate year 4 residents and five senior faculty members. The study was conducted with all phase III (Final year) students rotating in Gastroenterology. The residents and faculty members received brief training of one month in facilitation and core principles of adult education. Different aspects of teaching skills of residents and faculty were evaluated by students on a questionnaire (graded on Likert Scale from 1 to 10) assessing i) Knowledge Base-content Learning (KBL), ii) PBL, iii) Student Centered Learning (SCL) and iv) Group Skills (GS). There were 33 PBL teaching sessions in which 120 evaluation forms were filled; out of these 53% forms were filled for residents and 47% for faculty group. The faculty showed a statistically greater rating in "KBL" (faculty 8.37 Vs resident 7.94; p-value 0.02), "GS" (faculty 8.06 vs. residents 7.68; p-value 0.04). Differences in faculty and resident scores in "the PBL" and "SCL" were not significant. The overall score of faculty facilitators, however, was statistically significant for resident facilitators. (p = .05). 1) Residents are an effective supplement to faculty members for PBL; 2) Additional facilitators for PBL sessions can be identified in an institution by involvement of residents in teacher training workshops.


Biomechanical analysis of an expandable lateral cage and a static transforaminal lumbar interbody fusion cage with posterior instrumentation in an in vitro spondylolisthesis model.
Insufficient biomechanical data exist from comparisons of the stability of expandable lateral cages with that of static transforaminal lumbar interbody fusion (TLIF) cages. The purpose of this biomechanical study was to compare the relative rigidity of L4-5 expandable lateral interbody constructs with or without additive pedicle screw fixation with that of L4-5 static TLIF cages in a novel cadaveric spondylolisthesis model. Eight human cadaver spines were used in this study. A spondylolisthesis model was created at the L4-5 level by creating 2 injuries. First, in each cadaver, a nucleotomy from 2 channels through the anterior side was created. Second, the cartilage of the facet joint was burred down to create a gap of 4 mm. Light-emitting-diode tracking markers were placed at L-3, L-4, L-5, and S-1. Specimens were tested in the following scenarios: intact model, bilateral pedicle screws, expandable lateral 18-mm-wide cage (alone, with unilateral pedicle screws [UPSs], and with bilateral pedicle screws [BPSs]), expandable lateral 22-mm-wide cage (alone, with UPSs, and with BPSs), and TLIF (alone, with UPSs, and with BPSs). Four of the spines were tested with the expandable lateral cages (18-mm cage followed by the 22-mm cage), and 4 of the spines were tested with the TLIF construct. All these constructs were tested in flexion-extension, axial rotation, and lateral bending. The TLIF-alone construct was significantly less stable than the 18- and 22-mm-wide lateral lumbar interbody fusion (LLIF) constructs and the TLIF constructs with either UPSs or BPSs. The LLIF constructs alone were significantly less stable than the TLIF construct with BPSs. However, there was no significant difference between the 18-mm LLIF construct with UPSs and the TLIF construct with BPSs in any of the loading modes. Expandable lateral cages with UPSs provide stability equivalent to that of a TLIF construct with BPSs in a degenerative spondylolisthesis model.


Ising ferromagnet in dimension five: link and spin overlaps.
In the simple [hyper]cubic five-dimension, near-neighbor-interaction Ising ferromagnet, extensive simulation measurements are made of the link overlap and the spin overlap distributions. These "two replica" measurements are standard in the spin glass context but are not usually recorded in ferromagnet simulations. The moments and moment ratios of these distributions (the variance, the kurtosis, and the skewness) show clear critical behaviors at the known ordering temperature of the ferromagnet. Analogous overlap effects can be expected quite generally in Ising ferromagnets in any dimension. The link overlap results in particular, with peaks at criticality in the kurtosis and the skewness, also have implications for spin glasses.


Control of phonon transport by the formation of the Al2O3 interlayer in Al2O3-ZnO superlattice thin films and their in-plane thermoelectric energy generator performance.
Recently, significant progress has been made in increasing the figure-of-merit (ZT) of various nanostructured materials, including thin-film and quantum dot superlattice structures. Studies have focused on the size reduction and control of the surface or interface of nanostructured materials since these approaches enhance the thermopower and phonon scattering in quantum and superlattice structures. Currently, bismuth-tellurium-based semiconductor materials are widely employed for thermoelectric (TE) devices such as TE energy generators and coolers, in addition to other sensors, for use at temperatures under 400 K. However, new and promising TE materials with enhanced TE performance, including doped zinc oxide (ZnO) multilayer or superlattice thin films, are also required for designing solid-state TE power generating devices with the maximum output power density and for investigating the physics of in-plane TE generators. Herein, we report the growth of Al2O3/ZnO (AO/ZnO) superlattice thin films, which were prepared by atomic layer deposition (ALD), and the evaluation of their electrical and TE properties. All the in-plane TE properties, including the Seebeck coefficient (S), electrical conductivity (σ), and thermal conductivity (κ), of the AO/ZnO superlattice (with a 0.82 nm-thick AO layer) and AO/ZnO films (with a 0.13 nm-thick AO layer) were evaluated in the temperature range 40-300 K, and the measured S, σ, and κ were -62.4 and -17.5 μV K-1, 113 and 847 (Ω cm)-1, and 0.96 and 1.04 W m-1 K-1, respectively, at 300 K. Consequently, the in-plane TE ZT factor of AO/ZnO superlattice films was found to be ∼0.014, which is approximately two times more than that of AO/ZnO films (ZT of ∼0.007) at 300 K. Furthermore, the electrical power generation efficiency of the TE energy generator consisting of four couples of n-AO/ZnO superlattice films and p-Bi0.5Sb1.5Te3 (p-BST) thin-film legs on the substrate was demonstrated. Surprisingly, the output power of the 100 nm-thick n-AO/ZnO superlattice film/p-BST TE energy generator was determined to be ∼1.0 nW at a temperature difference of 80 K, corresponding to a significant improvement of ∼130% and ∼220% compared to the 100 nm-thick AO/ZnO film/p-BST and n-BT/p-BST film generators, respectively, owing to the enhancement of the TE properties, including the power factor of the superlattice film.


Identification and characterization of genes underlying chitinolysis in Collimonas fungivorans Ter331.
Through a combinatorial approach of plasposon mutagenesis, genome mining, and heterologous expression, we identified genes contributing to the chitinolytic phenotype of bacterium Collimonas fungivorans Ter331. One of five mutants with abolished ability to hydrolyze colloidal chitin carried its plasposon in the chiI gene coding for an extracellular endochitinase. Two mutants were affected in the promoter of chiP-II coding for an outer-membrane transporter of chitooligosaccharides. The remaining two mutations were linked to chitobiose/N-acetylglucosamine uptake. Thus, our model for the Collimonas chitinolytic system assumes a positive feedback regulation of chitinase activity by chitin degradation products. A second chitinase gene, chiII, coded for an exochitinase that preferentially released chitobiose from chitin analogs. Genes hexI and hexII showed coding resemblance to N-acetylglucosaminidases, and the activity of purified HexI protein towards chitin analogs suggested its role in converting chitobiose to N-acetylglucosamine. The hexI gene clustered with chiI, chiII, and chiP-II in one locus, while chitobiose/N-acetylglucosamine uptake genes colocalized in another. Both loci contained genes for conversion of N-acetylglucosamine to fructose-6-phosphate, confirming that C. fungivorans Ter331 features a complete chitin pathway. No link could be established between chitinolysis and antifungal activity of C. fungivorans Ter331, suggesting that the bacterium's reported antagonism towards fungi relies on other mechanisms.


Trapping and sensing 10 nm metal nanoparticles using plasmonic dipole antennas.
The optical trapping of Au nanoparticles with dimensions as small as 10 nm in the gap of plasmonic dipole antennas is demonstrated. Single nanoparticle trapping events are recorded in real time by monitoring the Rayleigh scattering spectra of individual plasmonic antennas. Numerical simulations are also performed to interpret the experimental results, indicating the possibility to trap nanoparticles only a few nanometers in size. This work unveils the potential associated with the integration of plasmonic trapping with localized surface plasmon resonance based sensing techniques, in order to deliver analyte to specific, highly sensitive regions ("hot spots").


An NGF-TrkA-mediated retrograde signal to transcription factor CREB in sympathetic neurons.
Nerve growth factor (NGF) is a neurotrophic factor secreted by cells that are the targets of innervation of sympathetic and some sensory neurons. However, the mechanism by which the NGF signal is propagated from the axon terminal to the cell body, which can be more than 1 meter away, to influence biochemical events critical for growth and survival of neurons has remained unclear. An NGF-mediated signal transmitted from the terminals and distal axons of cultured rat sympathetic neurons to their nuclei regulated phosphorylation of the transcription factor CREB (cyclic adenosine monophosphate response element-binding protein). Internalization of NGF and its receptor tyrosine kinase TrkA, and their transport to the cell body, were required for transmission of this signal. The tyrosine kinase activity of TrkA was required to maintain it in an autophosphorylated state upon its arrival in the cell body and for propagation of the signal to CREB within neuronal nuclei. Thus, an NGF-TrkA complex is a messenger that delivers the NGF signal from axon terminals to cell bodies of sympathetic neurons.


Generation of Anti-Boa Immunoglobulin Antibodies for Serodiagnostic Applications, and Their Use to Detect Anti-Reptarenavirus Antibodies in Boa Constrictor.
Immunoglobulins (Igs), the key effectors of the adaptive immune system, mediate the specific recognition of foreign structures, i.e. antigens. In mammals, IgM production commonly precedes the production of IgG in the response to an infection. The reptilian counterpart of IgG is IgY, but the exact kinetics of the reptilian immune response are less well known. Boid inclusion body disease (BIBD), an often fatal disease of captive boas and pythons has been linked to reptarenavirus infection, and BIBD is believed to be immunosuppressive. However, so far, the study of the serological response towards reptarenaviruses in BIBD has been hampered by the lack of reagents. Thus we set up a purification protocol for boa constrictor IgY and IgM, which should also be applicable for other snake species. We used centrifugal filter units, poly ethylene glycol precipitation and gel permeation chromatography to purify and separate the IgM and IgY fractions from boa constrictor serum, which we further used to immunise rabbits. We affinity purified IgM and IgY specific reagents from the produced antiserum, and labelled the reagents with horseradish peroxidase. Finally, using the sera of snakes with known exposure to reptarenaviruses we demonstrated that the newly generated reagents can be utilised for serodiagnostic purposes, such as immunoblotting and immunofluorescent staining. To our knowledge, this is the first report to show reptarenavirus-specific antibodies in boa constrictors.


Insulin-resistance and metabolic syndrome are related to executive function in women in a large family-based study.
While type 2 diabetes is well-known to be associated with poorer cognitive performance, few studies have reported on the association of metabolic syndrome (MetS) and contributing factors, such as insulin-resistance (HOMA-IR), low adiponectin-, and high C-reactive protein (CRP)-levels. We studied whether these factors are related to cognitive function and which of the MetS components are independently associated. The study was embedded in an ongoing family-based cohort study in a Dutch population. All participants underwent physical examinations, biomedical measurements, and neuropsychological testing. Linear regression models were used to determine the association between MetS, HOMA-IR, adiponectin levels, CRP, and cognitive test scores. Cross-sectional analyses were performed in 1,898 subjects (mean age 48 years, 43% men). People with MetS had significantly higher HOMA-IR scores, lower adiponectin levels, and higher CRP levels. MetS and high HOMA-IR were associated with poorer executive function in women (P = 0.03 and P = 0.009). MetS and HOMA-IR are associated with poorer executive function in women.


Changes in infant visual attention when observing repeated actions.
Infants' early visual preferences for faces, and their observational learning abilities, are well-established in the literature. The current study examines how infants' attention changes as they become increasingly familiar with a person and the actions that person is demonstrating. The looking patterns of 12- (n = 61) and 16-month-old infants (n = 29) were tracked while they watched videos of an adult presenting novel actions with four different objects three times. A face-to-action ratio in visual attention was calculated for each repetition and summarized as a mean across all videos. The face-to-action ratio increased with each action repetition, indicating that there was an increase in attention to the face relative to the action each additional time the action was demonstrated. Infant's prior familiarity with the object used was related to face-to-action ratio in 12-month-olds and initial looking behavior was related to face-to-action ratio in the whole sample. Prior familiarity with the presenter, and infant gender and age, were not related to face-to-action ratio. This study has theoretical implications for face preference and action observations in dynamic contexts.


Geographic bias in citation rates of conservation research.
We investigated whether the impact of conservation science is greater for research conducted in countries with more pressing conservation problems. We quantified research impact for 231 countries based on 2 citation metrics (mean cites per paper and h index) and fitted models predicting research impact based on number of threatened bird and mammal species (as a measure of conservation importance of a country) and a range of demographic variables. Citation rates of conservation research increased as a country's conservation need increased and as human population, quality of governance, and wealth increased. Even after accounting for these factors, citation rates among regions and countries within regions varied significantly. The conservation research community needs to consider ways to begin addressing the entrenched disadvantages some countries have when it comes to initiating projects and producing high-quality research.


Palladium-catalyzed intramolecular reductive olefin hydrocarbonation: benzylic hydrogen serving as a new hydrogen donor.
A palladium-catalyzed intramolecular hydrocarbonation of unactivated alkenes was achieved, in which toluene is used as a hydrogen donor for the first time. The radical transfer hydrogenation is designed and realized based on the Bond Dissociation Energy (BDE) value. A toluene derivative, which is cheap, readily available and easy to handle, serves as a new hydrogen donor in radical involved reactions, providing a novel and promising perspective for future reductive reactions.


A Live Video Program to Prevent Chronic Pain and Disability in At-Risk Adults With Acute Orthopedic Injuries (Toolkit for Optimal Recovery): Protocol for a Multisite Feasibility Study.
Despite the pivotal role of psychosocial factors in pain and disability after orthopedic injury, there are no evidence-based preventive interventions targeting psychosocial factors in patients with acute orthopedic injuries. We developed the first mind-body intervention focused on optimizing recovery and improving pain and disability in patients with acute orthopedic injuries who exhibit high levels of catastrophic thinking about pain and/or pain anxiety (Toolkit for Optimal Recovery [TOR] after orthopedic injury). In a pilot single-site randomized controlled trial (RCT), the TOR met a priori set benchmarks for feasibility, acceptability, and satisfaction. The next step in developing TOR is to conduct a multisite feasibility RCT to set the stage for a scientifically rigorous hybrid efficacy-effectiveness trial. The objective of this study is to conduct a rigorous multisite feasibility RCT of TOR to determine whether the intervention and study methodology meet a priori set benchmarks necessary for the successful implementation of a future multisite hybrid efficacy-effectiveness trial. In this paper, we describe the study design, manualized treatments, and specific strategies used to conduct this multisite feasibility RCT investigation. This study will be conducted at 3 geographically diverse level 1 trauma centers, anonymized as sites A, B, and C. We will conduct a multisite feasibility RCT of TOR versus the minimally enhanced usual care (MEUC) control (60 patients per site; 30 per arm) targeting a priori set feasibility benchmarks. Adult patients with acute orthopedic injuries who endorse high pain catastrophizing or pain anxiety will be recruited approximately 1-2 months after injury or surgery (baseline). Participants randomized to the TOR will receive a 4-session mind-body treatment delivered via a secure live video by trained clinical psychologists. Participants randomized to the MEUC will receive an educational booklet. Primary outcomes include feasibility of recruitment, appropriateness, feasibility of data collection, acceptability of TOR (adherence to sessions), and treatment satisfaction across all sites. We will also collect data on secondary implementation outcomes, as well as pain severity, physical and emotional function, coping skills, and adverse events. Outcomes will be assessed at baseline, posttreatment, and at the 3-month follow-up. Enrollment for the RCT is estimated to begin in June 2021. The target date of completion of the feasibility RCT is April 2024. The institutional review board approval has been obtained (January 2020). This investigation examines the multisite feasibility of TOR administered via live videoconferencing in adult patients with acute orthopedic injuries. If feasible, the next step is a multisite, hybrid efficacy-effectiveness trial of TOR versus MEUC. Preventive psychosocial interventions can provide a new way to improve patient and provider satisfaction and decrease suffering and health care costs among patients with orthopedic injuries who are at risk for chronic pain and disability. PRR1-10.2196/28155.


Commensal microbe-derived butyrate induces the differentiation of colonic regulatory T cells.
Gut commensal microbes shape the mucosal immune system by regulating the differentiation and expansion of several types of T cell. Clostridia, a dominant class of commensal microbe, can induce colonic regulatory T (Treg) cells, which have a central role in the suppression of inflammatory and allergic responses. However, the molecular mechanisms by which commensal microbes induce colonic Treg cells have been unclear. Here we show that a large bowel microbial fermentation product, butyrate, induces the differentiation of colonic Treg cells in mice. A comparative NMR-based metabolome analysis suggests that the luminal concentrations of short-chain fatty acids positively correlates with the number of Treg cells in the colon. Among short-chain fatty acids, butyrate induced the differentiation of Treg cells in vitro and in vivo, and ameliorated the development of colitis induced by adoptive transfer of CD4(+) CD45RB(hi) T cells in Rag1(-/-) mice. Treatment of naive T cells under the Treg-cell-polarizing conditions with butyrate enhanced histone H3 acetylation in the promoter and conserved non-coding sequence regions of the Foxp3 locus, suggesting a possible mechanism for how microbial-derived butyrate regulates the differentiation of Treg cells. Our findings provide new insight into the mechanisms by which host-microbe interactions establish immunological homeostasis in the gut.


Health & Demographic Surveillance System Profile: The Nairobi Urban Health and Demographic Surveillance System (NUHDSS).
The Nairobi Urban Health and Demographic Surveillance System (NUHDSS) was the first urban-based longitudinal health and demographic surveillance platform in sub-Saharan Africa (SSA). The NUHDSS was established in 2002 to provide a platform to investigate the long-term social, economic and health consequences of urban residence, and to serve as a primary research tool for intervention and impact evaluation studies focusing on the needs of the urban poor in SSA. Since its inception, the NUHDSS has successfully followed every year a population of about 65,000 individuals in 24,000 households in two slum communities--Korogocho and Viwandani--in Nairobi, Kenya. Data collected include key demographic and health information (births, deaths including verbal autopsy, in- and out-migration, immunization) and other information that characterizes living conditions in the slums (livelihood opportunities, household amenities and possessions, type of housing etc.). In addition to the routine data, it has provided a robust platform for nesting several studies examining the challenges of rapid urbanization in SSA and associated health and poverty dynamics. NUHDSS data are shared through internal and external collaborations, in accordance with the Centre's guidelines for publications, data sharing.


Anti-prothrombin autoantibodies enriched after infection with SARS-CoV-2 and influenced by strength of antibody response against SARS-CoV-2 proteins.
Antiphospholipid antibodies (aPL), assumed to cause antiphospholipid syndrome (APS), are notorious for their heterogeneity in targeting phospholipids and phospholipid-binding proteins. The persistent presence of Lupus anticoagulant and/or aPL against cardiolipin and/or β2-glycoprotein I have been shown to be independent risk factors for vascular thrombosis and pregnancy morbidity in APS. aPL production is thought to be triggered by-among other factors-viral infections, though infection-associated aPL have mostly been considered non-pathogenic. Recently, the potential pathogenicity of infection-associated aPL has gained momentum since an increasing number of patients infected with Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has been described with coagulation abnormalities and hyperinflammation, together with the presence of aPL. Here, we present data from a multicentric, mixed-severity study including three cohorts of individuals who contracted SARS-CoV-2 as well as non-infected blood donors. We simultaneously measured 10 different criteria and non-criteria aPL (IgM and IgG) by using a line immunoassay. Further, IgG antibody response against three SARS-CoV-2 proteins was investigated using tripartite automated blood immunoassay technology. Our analyses revealed that selected non-criteria aPL were enriched concomitant to or after an infection with SARS-CoV-2. Linear mixed-effects models suggest an association of aPL with prothrombin (PT). The strength of the antibody response against SARS-CoV-2 was further influenced by SARS-CoV-2 disease severity and sex of the individuals. In conclusion, our study is the first to report an association between disease severity, anti-SARS-CoV-2 immunoreactivity, and aPL against PT in patients with SARS-CoV-2.


Neuroanatomical correlates of low body weight in Alzheimer's disease: a PET study.
Weight loss is a common problem in patients with Alzheimer's disease (AD) and this might be associated with an increased risk for mortality. Recent evidences have suggested that certain brain dysfunctions may result in impaired nutritional status in AD patients. However, the mechanism of body weight loss in AD remains enigmatic. To investigate a possible association between low body weight and regional brain dysfunction, the authors conducted the correlational analysis of body mass index (BMI) with regional brain glucose metabolism using positron emission tomography (PET) and [18F]fluorodeoxyglucose (FDG). Twenty-seven patients with probable AD were divided into two groups: the low BMI group and the normal BMI group. Regional brain glucose metabolic ratios were calculated using the cerebellar hemisphere as a reference region. Comparisons were made of regional brain metabolic ratios between the low BMI group and the normal BMI group. Correlations of the glucose metabolic ratio with BMI were also assessed in all patients with AD. In the comparison between the two groups, glucose metabolic ratio in the anterior cingulate cortex (ACC) was significantly lower in the low BMI group, whereas no significant differences were found in all the other brain regions. Furthermore, regional glucose metabolism in the ACC had a significant and positive correlation with BMI (r=.450, P=.018). After adjustment for age, gender and disease duration, regional glucose metabolism in the ACC was independently associated with BMI. Our findings suggest that the ACC may be preferentially involved in the regulation of nutritional status in AD patients, and provides a new insight into developing strategies for prevention and treatment of undernutritional demented patients.


[Complete recovery of hungry bone syndrome using intravenous calcium infusion. Report of one case].
We report a 29 years old woman with a highly symptomatic primary hyperparathyroidism. After parathyroid adenoma excision, she presented a prolonged and life threatening hypocalcemia, due to a severe hungry bone syndrome. Conventional treatment with oral and intravenous calcium and calcitriol supplementation failed to raise serum and urinary calcium or to relief symptoms. After one month, we indicated a continuous intravenous calcium infusion allowing, during 6 months, an adequate outpatient management. Initial T scores for bone density were markedly low (L2-L4: -3.14; femoral neck: -3.07) and they increased 17% after 18 days of calcium infusion. After 147 days of treatment bone density was normal, increasing by 61%. The present case shows that the hungry bone syndrome can be a real risk for patients and a complex therapeutic challenge. With an appropriate calcium supply an early, fast and complete recovery of bone mass can be achieved.


Impact of mutations in Toll-like receptor pathway genes on esophageal carcinogenesis.
Esophageal adenocarcinoma (EAC) develops in an inflammatory microenvironment with reduced microbial diversity, but mechanisms for these influences remain poorly characterized. We hypothesized that mutations targeting the Toll-like receptor (TLR) pathway could disrupt innate immune signaling and promote a microenvironment that favors tumorigenesis. Through interrogating whole genome sequencing data from 171 EAC patients, we showed that non-synonymous mutations collectively affect the TLR pathway in 25/171 (14.6%, PathScan p = 8.7x10-5) tumors. TLR mutant cases were associated with more proximal tumors and metastatic disease, indicating possible clinical significance of these mutations. Only rare mutations were identified in adjacent Barrett's esophagus samples. We validated our findings in an external EAC dataset with non-synonymous TLR pathway mutations in 33/149 (22.1%, PathScan p = 0.05) tumors, and in other solid tumor types exposed to microbiomes in the COSMIC database (10,318 samples), including uterine endometrioid carcinoma (188/320, 58.8%), cutaneous melanoma (377/988, 38.2%), colorectal adenocarcinoma (402/1519, 26.5%), and stomach adenocarcinoma (151/579, 26.1%). TLR4 was the most frequently mutated gene with eleven mutations in 10/171 (5.8%) of EAC tumors. The TLR4 mutants E439G, S570I, F703C and R787H were confirmed to have impaired reactivity to bacterial lipopolysaccharide with marked reductions in signaling by luciferase reporter assays. Overall, our findings show that TLR pathway genes are recurrently mutated in EAC, and TLR4 mutations have decreased responsiveness to bacterial lipopolysaccharide and may play a role in disease pathogenesis in a subset of patients.


Novel monitor paradigm for real-time exposure assessment.
A wearable monitor that can reliably, accurately, and continuously measure personal exposure levels of various toxicants would not only accelerate the current environmental and occupational health and safety studies, but also enable new studies that are not possible with the current monitoring technology. Developing such a monitor has been a difficult challenge, and requires innovative sensing science and creative engineering. We have developed, built, and tested a wearable monitor for real-time detection of toxic hydrocarbons and acids in the environment. The monitor is low-cost, accurate, and user friendly. In addition, it can communicate wirelessly with a cell phone in which the monitoring results can be processed, displayed, stored, and transmitted to a designated computer. We have validated the functions and performance of the monitor, and carried out field tests with workers involving waste management, fire overhaul, and floor-cleaning activities, as well as with first- and second-hand smokers. The averaged exposure levels are in agreement with those determined by the standard NIOSH methods. The monitor provides accurate and real-time exposure assessment for the workers involving different activities. The real-time and continuous monitoring capability makes it possible to correlate the exposure levels with different activities and changes in the microenvironments. The monitor provides unprecedented real-time information that will help advance occupational safety and environmental health studies. It may also be used to better protect workers from occupational overexposure to toxic molecules.


Associations Between Non-neurological Autoimmune Disorders and Psychosis: A Meta-analysis.
A relationship between non-neurological autoimmune (NNAI) disorders and psychosis has been widely reported but not yet subjected to meta-analysis. We conducted the first meta-analysis examining the association between NNAI disorders and psychosis and investigated the effect of 1) temporality (as determined by study design), 2) psychiatric diagnosis, and 3) specific autoimmune disorders. Major databases were searched for articles published until April 2018; 31 studies, comprising data for >25 million individuals, were eligible. Using random-effects models, we examined the overall association between all NNAI disorders and psychosis; rheumatoid arthritis was examined separately given the well-established negative association with psychosis. Stratified analyses investigated the effect of temporality, psychiatric diagnosis, and specific NNAI disorders. We observed a positive overall association between NNAI disorders and psychosis (odds ratio [OR] = 1.26; 95% confidence interval [CI], 1.12-1.41) that was consistent across study designs and psychiatric diagnoses; however, considerable heterogeneity was detected (I2 = 88.08). Patterns varied across individual NNAI disorders; associations were positive for pernicious anemia (OR = 1.91; 95% CI, 1.29-2.84), pemphigoid (OR = 1.90; 95% CI, 1.62-2.24), psoriasis (OR = 1.70; 95% CI, 1.51-1.91), celiac disease (OR = 1.53; 95% CI, 1.12-2.10), and Graves' disease (OR = 1.33; 95% CI, 1.03-1.72) and negative for ankylosing spondylitis (OR = 0.72; 95% CI, 0.54-0.98) and rheumatoid arthritis (OR = 0.65; 95% CI, 0.50-0.84). While we observed a positive overall association between NNAI disorders and psychosis, this was not consistent across all NNAI disorders. Specific factors, including distinct inflammatory pathways, genetic influences, autoantibodies targeting brain proteins, and exposure to corticosteroid treatment, may therefore underlie this association.


CLIP-50 immunolocalization during mouse spermiogenesis suggests a role in shaping the sperm nucleus.
The spermatid nucleus and cytoplasm undergo dramatic morphological modifications during spermatid differentiation into mature sperm. Some of the external force causing this nuclear shaping is generated by a microtubular structure termed the manchette, which attaches to the perinuclear ring of the spermatid. Here, we report the isolation and characterization of a protein component of this perinuclear ring in an immunological screening of a mouse testis cDNA library. We termed this protein CLIP-50 because of its high similarity at the amino acid level to the C-terminal region of the microtubule-binding protein CLIP-170/restin. CLIP-50 lacks the characteristic microtubule-binding motif, but retains a portion of the predicted coiled-coiled domain and the metal-binding motif. The CLIP-50 transcript and protein are abundant in testis. The protein is also expressed in heart, lung, kidney, and skin. A distinct size variant exists in brain. In the spermatids, CLIP-50 protein localizes specifically to the centriolar region where the sperm tail originates and to the perinuclear ring from which the manchette emerges. CLIP-50 staining is retained in the ring throughout its migration over the surface of the nucleus which accompanies the nuclear shaping into its characteristic sperm configuration. This localization pattern indicates a very specific function for this novel CLIP derivative during mouse spermiogenesis.


Efficacy of chiropractic manual therapy on infant colic: a pragmatic single-blind, randomized controlled trial.
The purpose of this study was to determine the efficacy of chiropractic manual therapy for infants with unexplained crying behavior and if there was any effect of parental reporting bias. Infants with unexplained persistent crying (infant colic) were recruited between October 2007 and November 2009 at a chiropractic teaching clinic in the United Kingdom. Infants younger than 8 weeks were randomized to 1 of 3 groups: (i) infant treated, parent aware; (ii) infant treated, parent unaware; and (iii) infant not treated, parent unaware. The primary outcome was a daily crying diary completed by parents over a period of 10 days. Treatments were pragmatic, individualized to examination findings, and consisted of chiropractic manual therapy of the spine. Analysis of covariance was used to investigate differences between groups. One hundred four patients were randomized. In parents blinded to treatment allocation, using 2 or less hours of crying per day to determine a clinically significant improvement in crying time, the increased odds of improvement in treated infants compared with those not receiving treatment were statistically significant at day 8 (adjusted odds ratio [OR], 8.1; 95% confidence interval [CI], 1.4-45.0) and at day 10 (adjusted OR, 11.8; 95% CI, 2.1-68.3). The number needed to treat was 3. In contrast, the odds of improvement in treated infants were not significantly different in blinded compared with nonblinded parents (adjusted ORs, 0.7 [95% CI, 0.2-2.0] and 0.5 [95% CI, 0.1-1.6] at days 8 and 10, respectively). In this study, chiropractic manual therapy improved crying behavior in infants with colic. The findings showed that knowledge of treatment by the parent did not appear to contribute to the observed treatment effects in this study. Thus, it is unlikely that observed treatment effect is due to bias on the part of the reporting parent.


Signals on the move: chemokine receptors and organogenesis in zebrafish.
The chemokine SDF1 (stromal cell-derived factor 1) directs cell migration in many different contexts, ranging from embryogenesis to inflammation. SDF1a is the guidance cue for the zebrafish lateral line primordium, a tissue that moves along the flank of the embryo and deposits cells that form mechanosensory organs. The SDF1a receptor CXCR4b acts in cells at the leading edge of the primordium to direct its migration. Two new studies show that a second SDF1 receptor, CXCR7, is required only in the trailing cells of the primordium, and they explore how these two receptors orchestrate migration of the primordium. CXCR4b and CXCR7 are expressed in complementary domains, possibly through mutual repression in which each receptor inhibits expression of the other. These studies illustrate how the entire primordium can respond to a single signal, yet generate cell type-specific responses by using different receptors.


Audio computer-assisted self interview compared to traditional interview in an HIV-related behavioral survey in Vietnam.
Globally, population surveys on HIV/AIDS and other sensitive topics have been using audio computer-assisted self interview for many years. This interview technique, however, is still new to Vietnam and little is known about its application and impact in general population surveys. One plausible hypothesis is that residents of Vietnam interviewed using this technique may provide a higher response rate and be more willing to reveal their true behaviors than if interviewed with traditional methods. This study aims to compare audio computer-assisted self interview with traditional face-to-face personal interview and self-administered interview with regard to rates of refusal and affirmative responses to questions on sensitive topics related to HIV/AIDS. In June 2010, a randomized study was conducted in three cities (Ha Noi, Da Nan and Can Tho), using a sample of 4049 residents aged 15 to 49 years. Respondents were randomly assigned to one of three interviewing methods: audio computer-assisted self interview, personal face-to-face interview, and self-administered paper interview. Instead of providing answers directly to interviewer questions as with traditional methods, audio computer-assisted self-interview respondents read the questions displayed on a laptop screen, while listening to the questions through audio headphones, then entered responses using a laptop keyboard. A MySQL database was used for data management and SPSS statistical package version 18 used for data analysis with bivariate and multivariate statistical techniques. Rates of high risk behaviors and mean values of continuous variables were compared for the three data collection methods. Audio computer-assisted self interview showed advantages over comparison techniques, achieving lower refusal rates and reporting higher prevalence of some sensitive and risk behaviors (perhaps indication of more truthful answers). Premarital sex was reported by 20.4% in the audio computer-assisted self-interview survey group, versus 11.4% in the face-to-face group and 11.1% in the self-administered paper questionnaire group. The pattern was consistent for both male and female respondents and in both urban and rural settings. Men in the audio computer-assisted self-interview group also reported higher levels of high-risk sexual behavior--such as sex with sex workers and a higher average number of sexual partners--than did women in the same group. Importantly, item refusal rates on sensitive topics tended to be lower with audio computer-assisted self interview than with the other two methods. Combined with existing data from other countries and previous studies in Vietnam, these findings suggest that researchers should consider using audio computer-assisted self interview for future studies of sensitive and stigmatized topics, especially for men.


Molecular Basis for Differential Patterns of Drug Resistance in Influenza N1 and N2 Neuraminidase.
Neuraminidase (NA) inhibitors are used for the prevention and treatment of influenza A virus infections. Two subtypes of NA, N1 and N2, predominate in viruses that infect humans, but differential patterns of drug resistance have emerged in each subtype despite highly homologous active sites. To understand the molecular basis for the selection of these drug resistance mutations, structural and dynamic analyses on complexes of N1 and N2 NA with substrates and inhibitors were performed. Comparison of dynamic substrate and inhibitor envelopes and interactions at the active site revealed how differential patterns of drug resistance have emerged for specific drug resistance mutations, at residues I222, S246, and H274 in N1 and E119 in N2. Our results show that the differences in intermolecular interactions, especially van der Waals contacts, of the inhibitors versus substrates at the NA active site effectively explain the selection of resistance mutations in the two subtypes. Avoiding such contacts that render inhibitors vulnerable to resistance by better mimicking the dynamics and intermolecular interactions of substrates can lead to the development of novel inhibitors that avoid drug resistance in both subtypes.


EGFR Downregulation after Anti-EGFR Therapy Predicts the Antitumor Effect in Colorectal Cancer.
Anti-EGFR mAb is reported to induce EGFR internalization in colorectal cancer cells. However, the biological relevance of EGFR internalization with anti-EGFR mAb is unknown. Therefore, the relevance of EGFR downregulation with anti-EGFR mAb to antitumor activity in colorectal cancer cells was investigated. Quantification of EGFR on the cell surface before cetuximab treatment was assessed by flow cytometry, and its growth-inhibitory effects were measured by Trypan blue exclusion, in 10 RAS, BRAF wild-type colorectal cancer cell lines, but there was no significant correlation between EGFR number and its growth-inhibitory effect. However, a significant correlation existed between the percentage decrease in the number of EGFRs after cetuximab treatment and its growth-inhibitory effect in those cell lines. Treatment with TGFα, a ligand for EGFR, induced EGFR internalization in colorectal cancer cells, but most EGFRs subsequently recycled to the cell surface, consistent with previous studies. While cetuximab treatment induced EGFR internalization, most receptors subsequently translocated into the late endosome, leading to lysosomal degradation, as revealed by immunoblotting and double immunofluorescence. Cetuximab-sensitive colorectal cancer cells showed greater EGFR internalization, stronger cell growth inhibition, and more augmented apoptotic signals than nonsensitive cells. IHC for EGFR, performed using an EGFR pharmDx Kit (mouse anti-human EGFR mAb clone 2-18C9), in clinical specimens before and after anti-EGFR mAb therapy in 13 colorectal cancer patients showed a significant correlation between the response to anti-EGFR mAb and decreased staining after therapy.Implications: This report clearly demonstrates that anti-EGFR mAb facilitates internalization and subsequent degradation of EGFRs in lysosomes, which is an important determinant of the efficacy of anti-EGFR mAb treatment for colorectal cancer. Mol Cancer Res; 15(10); 1445-54. ©2017 AACR.


Test-retest Reliability and Correlations of 5 Global Measures Addressing At-work Productivity Loss in Patients with Rheumatic Diseases.
Several global measures to assess at-work productivity loss or presenteeism in patients with rheumatic diseases have been proposed, but the comparative validity is hampered by the lack of data on test-retest reliability and comparative concurrent and construct validity. Our objective was to test-retest 5 global measures of presenteeism and to compare the association between these scales and health-related well-being. Sixty-five participants with inflammatory arthritis or osteoarthritis in paid employment were recruited from 7 countries (UK, Canada, Netherlands, France, Sweden, Romania, and Italy). At baseline and 2 weeks later, 5 global measures of presenteeism were evaluated: the Work Productivity Scale-Rheumatoid Arthritis (WPS-RA), Work Productivity and Activity Impairment Questionnaire (WPAI), Work Ability Index (WAI), Quality and Quantity questionnaire (QQ), and the WHO Health and Performance Questionnaire (HPQ). Agreement between the 2 timepoints was assessed using single-measure intraclass correlations (ICC) and correlated between each other and with visual analog scale general well-being scores at followup by Spearman correlation. ICC between measures ranged from fair (HPQ 0.59) to excellent (WPS-RA 0.78). Spearman correlations between measures were moderate (Qquality vs WAI, r = 0.51) to strong (WPS-RA vs WPAI, r = 0.88). Correlations between measures and general well-being were low to moderate, ranging from -0.44 ≤ r ≤ 0.66. Test-retest results of 4 out of 5 global measures were good, and the correlations between these were moderate. The latter probably reflect differences in the concepts, recall periods, and references used in the measures, which implies that some measures are probably not interchangeable.


What is the effect of health coaching on physical activity participation in people aged 60 years and over? A systematic review of randomised controlled trials.
Physical inactivity is common in older age, yet increased activity benefits older people in terms of preventing chronic disease and maximising independence. Health coaching is a behaviour change intervention that has been shown to increase physical activity in clinical populations. This systematic review and meta-analysis investigated the effect of health coaching on physical activity, mobility, quality of life and mood in older people. MEDLINE, EMBASE, CENTRAL, PsycINFO, PEDro, SPORTDiscus, LILACS and CINAHL databases were used to identify randomised controlled trials which evaluated the effect of health coaching on physical activity (primary outcome) among people aged 60+. Secondary outcomes were mobility, quality of life and mood. We calculated standardised mean differences (SMDs, Hedges' g) with 95% CIs from random effects meta-analyses. 27 eligible trials were included. Health coaching had a small, statistically significant effect on physical activity (27 studies; SMD = 0.27; 95% CI 0.18 to 0.37; p<0.001). There was no evidence of an effect of health coaching on mobility (eight studies; SMD = 0.10; 95% CI -0.03 to 0.23; p=0.13), quality of life (eight studies; SMD = 0.07; 95% CI -0.06 to 0.20; p<0.05) or mood (five studies; SMD = 0.02; 95% CI -0.12 to 0.16; p=0.83). Health coaching significantly increased physical activity in people aged 60+. There was no evidence of an effect of health coaching on quality of life, mobility and mood, so different approaches may be required to impact on these outcomes.


A model for the evolution of prokaryotic DNA restriction-modification systems based upon the structural malleability of Type I restriction-modification enzymes.
Restriction Modification (RM) systems prevent the invasion of foreign genetic material into bacterial cells by restriction and protect the host's genetic material by methylation. They are therefore important in maintaining the integrity of the host genome. RM systems are currently classified into four types (I to IV) on the basis of differences in composition, target recognition, cofactors and the manner in which they cleave DNA. Comparing the structures of the different types, similarities can be observed suggesting an evolutionary link between these different types. This work describes the 'deconstruction' of a large Type I RM enzyme into forms structurally similar to smaller Type II RM enzymes in an effort to elucidate the pathway taken by Nature to form these different RM enzymes. Based upon the ability to engineer new enzymes from the Type I 'scaffold', an evolutionary pathway and the evolutionary pressures required to move along the pathway from Type I RM systems to Type II RM systems are proposed. Experiments to test the evolutionary model are discussed.


Modified roll technique for soft tissue augmentation in prosthetic rehabilitation: A case report.
There are various causes for residual alveolar ridge defects, most common being the collapse of the cortical plate during extraction. Localized defects of alveolar ridge affect prosthetic restoration due to poor emergence profile of the pontic. Various soft tissue augmentation procedures have been recommended for the correction of these defects. Abrams's roll technique has been widely used for soft tissue ridge augmentation in the maxillary anterior region where aesthetics is a prime concern. The original Abrams's roll technique has been modified and revisited by many authors to overcome the limitations. The present clinical case report discusses the modified roll technique as an option to treat alveolar ridge deformity. The current procedure resulted in expectable ridge augmentation, uneventful healing, and good aesthetic outcome. Hence this modified technique can be selected for the treatment of mild to moderate Seibert class I ridge defects in the aesthetic zone.


Laser in situ keratomileusis using the Nidek EC-5000 or the Alcon LADARVision 4000 systems.
To compare the refractive results of eyes with low to moderate myopia that underwent laser in situ keratomileusis with either the Nidek EC-5000 or the Alcon LADARVision 4000 excimer laser systems. We performed a retrospective review of 114 LASIK procedures with either the Nidek EC-5000 (54 eyes) or the Alcon LADARVision 4000 (60 eyes) excimer laser systems. Preoperative refractive errors were similar and both populations were treated during the same time period. Data analyzed included uncorrected visual acuity (UCVA), spherical magnitude, spherical equivalent refraction, astigmatism power, astigmatism axis, and vector astigmatism change. Results at 6 months were analyzed. Spherical correction change was a mean -3.95 D for Nidek treated eyes and a mean -4.53 D for LADARVision treated eyes (P = .20). Mean spherical equivalent refraction change was -3.70 D for Nidek eyes and -4.20 D for LADARVision eyes (P = .23). Mean change in UCVA (LogMAR) was 1.05 for Nidek eyes and 0.99 for LADARVision eyes (P = .40). Mean astigmatism magnitude change was 0.71 D for Nidek eyes and 0.77 D for LADARVision eyes (P = .63). Mean vector-corrected astigmatism change was 0.93 D for Nidek eyes and 1.00 D for LADARVision eyes (P = .63). Mean vector-corrected astigmatism axis for Nidek eyes was 3.08 D and for LADARVision eyes 6.58 D (P = .70). There was no significant difference in refractive results in eyes treated with the Alcon LADARVision 4000 or the Nidek EC-5000 excimer laser systems. Inherent differences between the two laser systems are highlighted.


Dielectrophoretic microfluidic device for the continuous sorting of Escherichia coli from blood cells.
Microfluidic diagnostic devices promise faster disease identification by purifying and concentrating low-abundance analytes from a flowing sample. The diagnosis of sepsis, a whole body inflammatory response often caused by microbial infections of the blood, is a model system for pursuing the advantages of microfluidic devices over traditional diagnostic protocols. Traditional sepsis diagnoses require large blood samples and several days to culture and identify the low concentration microbial agent. During these long delays while culturing, the physician has little or no actionable information to treat this acute illness. We designed a microfluidic chip using dielectrophoresis to sort and concentrate the target microbe from a flowing blood sample. This design was optimized using the applicable electrokinetic and hydrodynamic theories. We quantify the sorting efficiency of this device using growth-based assays which show 30% of injected microbes are recovered viable, consistent with the electroporation of target cells by the dielectrophoretic cell sorters. Finally, the results illustrate the device is capable of a five-fold larger microbe concentration in the target analyte stream compared to the waste stream at a continuous sample flow rate of 35 μl∕h.


A test of virtual auditory localization.
The purpose of this study was to evaluate a test of virtual auditory localization including assessment of its ease of administration and its sensitivity to differences in binaural performance in children and adults. This test eliminates many potential problems inherent in any free-field localization test such as calibration problems, problems replicating source and listener locations, and issues associated with head movements. Binaural performance was measured using the virtual localization test and a simple binaural detection task, the masking-level difference (MLD), for three groups of subjects: adults, children with a negative history of otitis media, and children with a positive history of otitis media. There were five subjects in each group. The adults were all student volunteers; the children were recruited first and subsequently placed into groups based on their medical histories obtained from their physicians and parental reports. Results indicate that this test of virtual auditory localization is useful for measuring binaural performance in children and adults and is sensitive to differences in binaural processing. Performance of the adults and children with a negative history of otitis media was comparable on both of the binaural tests, and on the binaural detection task, was similar to that reported in the literature for normal-hearing listeners; but the children with a positive history of otitis media performed more poorly on both tests. The results of this study indicate that the virtual localization test described here is easy to administer to children and adults. The signal processing techniques used in this virtual auditory localization test lend themselves to straightforward comparisons across different laboratories and clinics and make this test a potentially useful clinical tool. The development of such a clinical test is currently under study.


Survey on advance care planning of Italian outpatients on chronic haemodialysis.
The clinical practice guidelines published by the Renal Physicians Association (USA) recommend instituting advance care planning (ACP) for patients with end-stage renal disease. Studies on this issue are lacking in Italy. Our aim was to determine the attitudes of patients on ACP in our dialysis centre. We performed a cross-sectional survey. We recruited patients on maintenance haemodialysis (HD) at Hemodialysis Center of Università Cattolica del Sacro Cuore, from 1 March 2014 to 31 March 2015. The only exclusion criterion was inability to give an informed consent. Patients completed a questionnaire concerning their treatment preferences in three hypothetical disease scenarios: persistent vegetative state, advanced dementia, severe terminal illness; for each patients, we also collected clinical, functional and socioeconomic data. Thirty-four HD outpatients completed the study questionnaire. The majority of respondents (85%) considered information about prognosis, health conditions and treatment options, including withdrawing dialysis, as very important and 94% of respondents considered treatment of uraemic/dialytic symptoms the most important issue. In the health scenarios provided, dialysis was the treatment least withheld. Dependence on instrumental activities of daily living (0.048) and higher Charlson Comorbidity Index scores (p=0.035) were associated with continuing dialysis in at least one scenario. ACP should be tailored to patients' value, culture and preferences. A significant proportion of patients, however, do not want to be involved in end of life decisions. Frail elderly patients, in particular, are not inclined to interrupt dialysis, despite poor quality of life or a poor prognosis.


Functional disability, cognitive impairment, and depression after hospitalization for pneumonia.
The study objective was to examine whether hospitalization for pneumonia is associated with functional decline, cognitive impairment, and depression, and to compare this impairment with that seen after known disabling conditions, such as myocardial infarction or stroke. We used data from a prospective cohort of 1434 adults aged more than 50 years who survived 1711 hospitalizations for pneumonia, myocardial infarction, or stroke drawn from the Health and Retirement Study (1998-2010). Main outcome measures included the number of Activities and Instrumental Activities of Daily Living requiring assistance and the presence of cognitive impairment and substantial depressive symptoms. Hospitalization for pneumonia was associated with 1.01 new impairments in Activities and Instrumental Activities of Daily Living (95% confidence interval [CI], 0.71-1.32) among patients without baseline functional impairment and 0.99 new impairments in Activities and Instrumental Activities of Daily Living (95% CI, 0.57-1.41) among those with mild-to-moderate baseline limitations, as well as moderate-to-severe cognitive impairment (odds ratio, 2.46; 95% CI, 1.60-3.79) and substantial depressive symptoms (odds ratio, 1.63; 95% CI, 1.06-2.51). Patients without baseline functional impairment who survived pneumonia hospitalization had more subsequent impairments in Activities and Instrumental Activities of Daily Living than those who survived myocardial infarction hospitalization. There were no significant differences in subsequent moderate-to-severe cognitive impairment or substantial depressive symptoms between patients who survived myocardial infarction or stroke and those who survived pneumonia. Hospitalization for pneumonia in older adults is associated with subsequent functional and cognitive impairment. Improved pneumonia prevention and interventions to ameliorate adverse sequelae during and after hospitalization may improve outcomes.


Seroprevalence of Dengue Fever in US Army Special Operations Forces: Initial Results and the Way Ahead.
The endemicity of dengue fever (DF) and, consequently, sequelae of DF are increasing worldwide. The increases are largely a result of widespread international travel and the increased range of the mosquito vectors. US Army Special Operations Command (USASOC) personnel are at an increased risk of exposure to dengue based on their frequent deployments to and presence in dengue endemic areas worldwide. Repeated deployments to different endemic areas can increase the risk for developing the more serious sequelae of dengue: dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). Information about the seroprevalence rate of dengue in USASOC personnel, in particular, is lacking and is critical to assessing the risk, tailoring preventive medicine countermeasures, leveraging field diagnostics, and maintaining mission capability. In the first part of a two-part project to assess baseline seroprevalence in USASOC units, a random, unit-stratified sample of 500 anonymous serum specimens from personnel assigned to the highest-risk units in USASOC were screened for dengue using a microneutralization assay. Of the 500 specimens screened, 56 (11.2%) of 500 had neutralizing titers (NT) (MN₅₀≥10) against at least one DENV serotype. Subsequent sample titration resulted in 48 (85.7%) of 56 of the samples with NT (MN₅₀≥10) against at least one dengue serotype for an overall dengue exposure rate of 9.6% (48 of 500). The second part of the ongoing project, started in 2012, was a multicenter, serosurveillance project using predeployment and postdeployment sera collected from USASOC personnel deployed to South and Central America, Africa, and Southeast Asia. Preliminary results show a 13.2% (55 of 414) seropositivity rate. The significance of these findings as they relate to personal risk and operational impact is discussed.


Molecular genealogy tools for white-tailed deer with chronic wasting disease.
Molecular genetic data provide powerful tools for genealogy reconstruction to reveal mechanisms underlying disease ecology. White-tailed deer (Odocoileus virginianus) congregate in matriarchal groups; kin-related close social spacing may be a factor in the spread of infectious diseases. Spread of chronic wasting disease (CWD), a prion disorder of deer and their cervid relatives, is presumed to be associated with direct contact between individuals and by exposure to shared food and water sources contaminated with prions shed by infected deer. Key aspects of disease ecology are yet unknown. DNA tools for pedigree reconstruction were developed to fill knowledge gaps in disease dynamics in prion-infected wild animals. Kinship indices using data from microsatellite loci and sequence haplotypes of mitochondrial DNA were employed to assemble genealogies. Molecular genealogy tools will be useful for landscape-level population genetic research and monitoring, in addition to epidemiologic studies examining transmission of CWD in captive and free-ranging cervids.


Psychosocial rehabilitation activities, empowerment, and quality of community-based life for people with schizophrenia.
Many variables influencing quality of life (QOL) for outpatients with schizophrenia have been identified from prior studies. Symptom severity, psychosocial rehabilitation activities, and empowerment have all been clearly identified as key variables. However, which variables are the most influential and important factors remains unknown; factors influencing QOL, either directly or indirectly and to what degree, need to be examined. The aim of this study was to test the hypothesis that empowerment is a possible mediator of how (a) psychiatric symptoms and (b) psychosocial rehabilitation activities affect QOL for outpatients with schizophrenia in the community. We used the probability proportional to size random sampling for 190 outpatients with schizophrenia at 10 community rehabilitation centers in Taipei, such that samples consisted of adults who fulfilled the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition criteria. The instruments included the questionnaire to gather demographic and disease information, the Empowerment Scale, the Psychiatric Symptoms Scale, the psychosocial rehabilitation activity (PRA), and the Quality of Life Scale for Psychiatric Patients. Beyond descriptive statistics, correlation and structural equation models were computed. Findings showed that empowerment in outpatients with schizophrenia mediates QOL, whereas psychosocial rehabilitation activities seem to increase empowerment, which may in turn increase QOL. Psychotic symptoms seem to have a direct effect of decreasing QOL that could not be mediated by empowerment. Empowerment had a significant effect on QOL for outpatients with schizophrenia. The findings of this study support the importance of empowerment and rehabilitation activities for promoting QOL among community outpatients. We suggest that various rehabilitation programs and empowerment health education are needed to enhance QOL for schizophrenia outpatients in the community.


Acute Stroke Therapy by Inhibition of Neutrophils (ASTIN): an adaptive dose-response study of UK-279,276 in acute ischemic stroke.
UK-279,276 (neutrophil inhibitory factor) reduced infarct volume in a rat middle cerebral artery occlusion reperfusion model. ASTIN (Acute Stroke Therapy by Inhibition of Neutrophils) was an adaptive phase 2 dose-response-finding, proof-of-concept study to establish whether UK-279,276 improves recovery in acute ischemic stroke. The prime objective was to determine the dose that gave a clinically relevant effect in patients. A Bayesian sequential design with real-time efficacy data capture and continuous reassessment of the dose response allowed double-blind, randomized, adaptive allocation to 1 of 15 doses (dose range, 10 to 120 mg) or placebo and early termination for efficacy or futility. The primary end point was change from baseline to day 90 on the Scandinavian Stroke Scale (DeltaSSS), adjusted for baseline SSS, aiming for a 3-point additional mean recovery above placebo. Nine hundred sixty-six acute stroke patients (887 ischemic, 204 cotreated with intravenous tissue plasminogen activator; mean baseline SSS score, 28; range, 10 to 40) were treated within 6 hours of symptom onset. Mean DeltaSSS was approximately +17 points of improvement on SSS for the overall evaluable population. There was no treatment effect for UK-279,276 (posterior probability of futility, 0.89). The trial was stopped early for futility. Post hoc analysis indicated a mean 1.6-point additional improvement on DeltaSSS in the tissue plasminogen activator-treated subset (credible interval=0.5, 2.6). UK-279,276 was generally well tolerated, with no increased incidence of infections. UK-279,276 did not improve recovery in acute ischemic stroke patients but was devoid of serious side effects. The adaptive design facilitated early termination for futility.


[Association of lifestyle and cardiometabolic risk factors with epigenetic age acceleration in adults in China].
Objective: To explore the association of lifestyle and cardiometabolic risk factors with five epigenetic age acceleration (AA) indices. Methods: This study included 980 participants of China Kadoorie Biobank, for whom genome-wide DNA methylation of peripheral blood cells had been detected in baseline survey. Five indices of DNA methylation age (DNAm age) were calculated, i.e. Horvath clock, Hannum clock, DNAm PhenoAge, GrimAge and Li clock. Epigenetic AA was defined as the residual of regressing DNAm age on chronological age. Lifestyle factors studied included smoking status, alcohol consumption, eating habits, physical activity level and body shape defined by a combination of BMI and waist circumference. Cardiometabolic risk factors included blood pressure, blood glucose level and total cholesterol level. Linear regression model was used to analyze the association of lifestyle and cardiometabolic risk factors with AA (β). Results: GrimAge_AA was associated with smoking status, alcohol consumption, physical activity level and BMI. Compared with non-smokers, non-drinkers, or participants with BMI of 18.5- 23.9 kg/m2, the smokers who smoked 1-14 cigarettes/day (β=0.71, 95%CI: 0.57-0.86), 15-24 cigarettes/day (β=0.88, 95%CI: 0.73-1.03), and ≥25 cigarettes/day (β=0.99, 95%CI: 0.81-1.18), respectively, heavy drinkers with daily pure alcohol consumption ≥60 g (β=0.33, 95%CI: 0.11-0.55) and participants with BMI<18.5 kg/m2 (β=0.23, 95%CI: 0.03-0.43) showed accelerated aging. Compared with those in the lowest quintile of physical activity level, participants in the top three quintile of physical activity level showed decelerated aging (β=-0.13, 95%CI: -0.26-0.01, β=-0.12, 95%CI: -0.26-0.02, and β=-0.14, 95%CI: -0.27- -0.00, respectively). GrimAge_AA decreased with the increase of the number of healthy lifestyle factors (P<0.001). Compared with the participants with 0 to 1 healthy lifestyle factor, the β of those with 2, 3, or 4 to 5 healthy lifestyle factors were -0.30 (95%CI: -0.47- -0.12), -0.47 (95%CI: -0.65- -0.30) and -0.72 (95%CI: -0.90- -0.53), respectively. The other four indices were not statistically significantly associated with most lifestyle factors. None of the five indices of AA was associated with blood pressure, blood glucose level or total cholesterol level. Conclusion: People with unhealthy lifestyle showed accelerated epigenetic aging, that is, the predicted DNAm age is older than their own chronological age.


Molecular basis for the antiviral and anticancer activities of unnatural L-beta-nucleosides.
As a general rule, enzymes act on only one enantiomer of a chiral substrate and only one of the enantiomeric forms of a chiral molecule may bind effectively at the catalytic site, displaying biological activity. In recent years, some exceptions have been found among viral and cellular enzymes involved in the synthesis of deoxynucleoside triphosphates and in their polymerisation into DNA. Examples are: herpes virus thymidine kinases, cellular deoxycytidine kinase and deoxynucleotide kinases, human immunodeficiency virus type 1 (HIV-1) reverse transcriptase, hepatitis B virus (HBV) DNA polymerase and, to a lesser extent, some cellular DNA polymerases. The lack of enantioselectivity allows herpes simplex virus (HSV) thymidine kinase and cellular deoxycytidine kinase to phosphorylate the unnatural L-beta-enantiomers of D-thymidine and D-deoxycytidine, respectively, or of their analogues to monophosphate. This phosphorylation represents the first and often the rate-limiting step of their activation to triphosphates. The L-triphosphates can then exert antiviral (anti-HSV, anti-Human cytomegalovirus, anti-HIV-1, anti-HBV) and anticancer activities. Although only one L-nucleoside (3TC) has so far gained United States of America Food and Drug Administration (USA FDA) approval for clinical use against HIV-1, other L-enantiomers of nucleoside analogues, which have shown antiviral or anticancer activity in cell cultures are in clinical trials. Their resistance to enantioselective enzymes, such as thymidine phosphorylase, thymidylate synthase, (deoxy)-cytidine and dCMP deaminases, and their lower affinity for the mitochondrial thymidine kinase can ensure a higher selectivity and lower cytotoxicity with respect to those exerted by their corresponding natural D-enantiomers and might be exploited to solve problems arising during chemotherapy, such as metabolic inactivation, cytotoxicity and drug-resistance.


Tunable protein harmonic diffractive micro-optical elements.
Herein, tunable protein harmonic diffractive microlenses (PHDMs) have been constructed by femtosecond laser direct writing from bovine serum albumin. With excellent three-dimensional topography, PHDMs show distinct harmonic diffractive features, such as similarly good imaging performance with smaller thickness than refractive lenses and well controlled minus chromatic dispersion compared with simple diffractive lenses (∼5% focal length shift against ∼21% of the protein Fresnel zone plate with the same radius under light with wavelength 450-659 nm). Owing to the nature of protein molecules as "building blocks," the focal length of the PHDM can be facilely tuned within seconds by changing the pH value (focal length tunability of up to ∼20%).


Structure of the human κ-opioid receptor in complex with JDTic.
Opioid receptors mediate the actions of endogenous and exogenous opioids on many physiological processes, including the regulation of pain, respiratory drive, mood, and--in the case of κ-opioid receptor (κ-OR)--dysphoria and psychotomimesis. Here we report the crystal structure of the human κ-OR in complex with the selective antagonist JDTic, arranged in parallel dimers, at 2.9 Å resolution. The structure reveals important features of the ligand-binding pocket that contribute to the high affinity and subtype selectivity of JDTic for the human κ-OR. Modelling of other important κ-OR-selective ligands, including the morphinan-derived antagonists norbinaltorphimine and 5'-guanidinonaltrindole, and the diterpene agonist salvinorin A analogue RB-64, reveals both common and distinct features for binding these diverse chemotypes. Analysis of site-directed mutagenesis and ligand structure-activity relationships confirms the interactions observed in the crystal structure, thereby providing a molecular explanation for κ-OR subtype selectivity, and essential insights for the design of compounds with new pharmacological properties targeting the human κ-OR.


Differences in muscarinic receptor reserve for inhibition of adenylate cyclase and stimulation of phosphoinositide hydrolysis in chick heart cells.
Carbachol is 100 times more potent for inhibiting cyclic AMP formation than for stimulating phosphoinositide (PI) hydrolysis in chick heart cells. To determine whether this reflects differences in agonist affinity of the receptor(s) coupled to the two responses, we measured these functional responses following removal of receptor reserve with propylbenzilycholine mustard (PrBCM). Conditions of PrBCM treatment that led to progressive loss of up to 95% of the [3H]-N-methylscopolamine-binding sites decreased the potency but not the maximal capacity of carbachol to inhibit cyclic AMP formation. In contrast, there was a marked decrease in the maximal PI response to carbachol. The KA for carbachol, calculated by measuring functional responses following receptor inactivation, was similar whether the cyclic AMP or the PI response was examined. These KA values (approximately 40 microM) were similar to the KD calculated by examining carbachol competition for [3H]-N-methylscopolamine-binding sites on the intact cell. PrBCM treatment also decreased the maximal effect of oxotremorine on cyclic AMP formation under conditions in which carbachol remained a full agonist for this response. We interpret our data as indicating that: there is much greater receptor reserve in the coupling of muscarinic receptors to adenylate cyclase than to PI hydrolysis; this, rather than differences in receptor affinity underlies the disparate dose-response relationships for the two responses; and differences in the effects of weak agonist on the two responses may also reflect differences in receptor reserve. We suggest that muscarinic receptors with the same affinity for carbachol interact with different efficiency with the transducers (Gi and Gx) that regulate adenylate cyclase and phospholipase C.


Hierarchical Graphene-Encapsulated Hollow SnO2@SnS2 Nanostructures with Enhanced Lithium Storage Capability.
Complex hierarchical structures have received tremendous attention due to their superior properties over their constitute components. In this study, hierarchical graphene-encapsulated hollow SnO2@SnS2 nanostructures are successfully prepared by in situ sulfuration on the backbones of hollow SnO2 spheres via a simple hydrothermal method followed by a solvothermal surface modification. The as-prepared hierarchical SnO2@SnS2@rGO nanocomposite can be used as anode material in lithium ion batteries, exhibiting excellent cyclability with a capacity of 583 mAh/g after 100 electrochemical cycles at a specific current of 200 mA/g. This material shows a very low capacity fading of only 0.273% per cycle from the second to the 100th cycle, lower than the capacity degradation of bare SnO2 hollow spheres (0.830%) and single SnS2 nanosheets (0.393%). Even after being cycled at a range of specific currents varied from 100 mA/g to 2000 mA/g, hierarchical SnO2@SnS2@rGO nanocomposites maintain a reversible capacity of 664 mAh/g, which is much higher than single SnS2 nanosheets (374 mAh/g) and bare SnO2 hollow spheres (177 mAh/g). Such significantly improved electrochemical performance can be attributed to the unique hierarchical hollow structure, which not only effectively alleviates the stress resulting from the lithiation/delithiation process and maintaining structural stability during cycling but also reduces aggregation and facilitates ion transport. This work thus demonstrates the great potential of hierarchical SnO2@SnS2@rGO nanocomposites for applications as a high-performance anode material in next-generation lithium ion battery technology.


Lost in reciprocal space? Determination of the scattering condition in spot profile analysis low-energy electron diffraction.
The precise knowledge of the diffraction condition, i.e., the angle of incidence and electron energy, is crucial for the study of surface morphology through spot profile analysis low-energy electron diffraction (LEED). We demonstrate four different procedures to determine the diffraction condition: employing the distortion of the LEED pattern under large angles of incidence, the layer-by-layer growth oscillations during homoepitaxial growth, a G(S) analysis of a rough surface, and the intersection of facet rods with 3D Bragg conditions.


Dental hygienists' self-reported performance of tobacco cessation activities.
Tobacco use is a major risk factor in oral and systemic diseases. Current national guidelines outline activities that positively contribute towards tobacco cessation. Little is known about dental hygienists' role in tobacco cessation activities. This study investigates dental hygienists' performance of tobacco cessation activities in the dental surgery and explores factors associated with this performance. Dental hygienists in south-east England on the register of the British Society of Dental Hygiene and Therapy completed a postal questionnaire survey in 2008. A total of 439 (61%) questionnaires were returned, representing both part-time (66.7%) and full-time (33.3%) hygienists working in the private (69.5%) and non-private sectors (30.5%). Most hygienists recorded positive attitudes towards tobacco cessation. Overall, 162 (41.9%) reported performing four or more tobacco cessation activities. The less-commonly performed activities indicated to patients the value of attending NHS Stop Smoking Services for specialised help, with hygienists referring patients to the NHS Stop Smoking Services and offering assistance to those who do not wish to attend NHS stop Smoking Services. Higher reported numbers of tobacco cessation activities performed were statistically significantly associated with a higher number of positive attitudes held (P = 0.004), while socio-demographic and professional attributes were not. Tobacco cessation activities, as recommended by national guidelines, are not always performed by dental hygienists. Efforts are needed to promote the dental hygienist's role in tobacco cessation, especially in relation to referring patients to NHS Stop Smoking services. These efforts should focus on changing the attitudes of dental hygienists, especially on their effectiveness in promoting tobacco cessation.


Modeling tumor microenvironments in vitro.
Tumor progression depends critically upon the interactions between the tumor cells and their microenvironment. The tumor microenvironment is heterogeneous and dynamic; it consists of extracellular matrix, stromal cells, immune cells, progenitor cells, and blood and lymphatic vessels. The emerging fields of tissue engineering and microtechnologies have opened up new possibilities for engineering physiologically relevant and spatially well-defined microenvironments. These in vitro models allow specific manipulation of biophysical and biochemical parameters, such as chemical gradients, biomatrix stiffness, metabolic stress, and fluid flows; thus providing a means to study their roles in certain aspects of tumor progression such as cell proliferation, invasion, and crosstalk with other cell types. Challenges and perspectives for deconvolving the complexity of tumor microenvironments will be discussed. Emphasis will be given to in vitro models of tumor cell migration and invasion.