Regulation of mitochondrial oxidative metabolism by tumor suppressor FLCN.
Birt-Hogg-Dubé (BHD) syndrome is a hereditary hamartoma syndrome that predisposes patients to develop hair follicle tumors, lung cysts, and kidney cancer. Genetic studies of BHD patients have uncovered the causative gene, FLCN, but its function is incompletely understood. Mice with conditional alleles of FLCN and/or peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PPARGC1A), a transcriptional coactivator that regulates mitochondrial biogenesis, were crossbred with mice harboring either muscle creatine kinase (CKM) -Cre or myogenin (MYOG) -Cre transgenes to knock out FLCN and/or PPARGC1A in muscle, or cadherin 16 (CDH16)- Cre transgenes to knock out FLCN and/or PPARGC1A in kidney. Real-time polymerase chain reaction, immunoblotting, electron microscopy, and metabolic profiling assay were performed to evaluate mitochondrial biogenesis and function in muscle. Immunoblotting, electron microscopy, and histological analysis were used to investigate expression and the pathological role of PPARGC1A in FLCN-deficient kidney. Real-time polymerase chain reaction, oxygen consumption measurement, and flow cytometry were carried out using a FLCN-null kidney cancer cell line. All statistical analyses were two-sided. Muscle-targeted FLCN knockout mice underwent a pronounced metabolic shift toward oxidative phosphorylation, including increased mitochondrial biogenesis (FLCN ( f/f ) vs FLCN ( f/f ) /CKM-Cre: % mitochondrial area mean = 7.8% vs 17.8%; difference = 10.0%; 95% confidence interval = 5.7% to 14.3%; P < .001), and the observed increase in mitochondrial biogenesis was PPARGC1A dependent. Reconstitution of FLCN-null kidney cancer cells with wild-type FLCN suppressed mitochondrial metabolism and PPARGC1A expression. Kidney-targeted PPARGC1A inactivation partially rescued the enlarged kidney phenotype and abrogated the hyperplastic cells observed in the FLCN-deficient kidney. FLCN deficiency and subsequent increased PPARGC1A expression result in increased mitochondrial function and oxidative metabolism as the source of cellular energy, which may give FLCN-null kidney cells a growth advantage and drive hyperplastic transformation.


Identification of ethnomedicinal plants (Rauvolfioideae: Apocynaceae) through DNA barcoding from northeast India.
DNA barcode-based molecular characterization is in practice for plants, but yet lacks total agreement considering the selection of marker. Plant species of subfamily Rauvolfioideae have long been used as herbal medicine by the majority of tribal people in Northeast (NE) India and at present holds mass effect on the society. Hence, there is an urgent need of correct taxonomic inventorization vis-à-vis species level molecular characterization of important medicinal plants. To test the efficiency of matK in species delineation like DNA barcoding in Rauvolfiadae (Apocynaceae). In this study, the core DNA barcode matK and trnH-psbA sequences are examined for differentiation of selected ethnomedicinal plants of Apocynaceae. DNA from young leaves of selected species was isolated, and matK gene (~800 bp) and trnH-psbA spacer (~450 bp) of Chloroplast DNA was amplified for species level identification. The ~758 bp matK sequence in comparison to the trnH-psbA showed easy amplification, alignment, and high level of discrimination value among the medicinal Rauvolfioidae species. Intergenic spacer trnH-psbA is also exhibited persistent problem in obtaining constant bidirectional sequences. Partial matK sequences exhibited 3 indels in multiple of 3 at 5 end. Evidently, generated matK sequences are clustered cohesively, with their conspecific Genbank sequences. However, repeat structures with AT-rich regions, possessing indels in multiple of 3, could be utilized as qualitative molecular markers in further studies both at the intra-specific and shallow inter-specific levels like the intergenic spacers of CpDNA. matK sequence information could help in correct species identification for medicinal plants of Rauvolfioideae.


Essay On Being a Doctor: Monday Morning.
Through the generosity of Charles R. Millikan, DMin, vice president for Spiritual Care and Values Integration, an annual award competition was established at Houston Methodist Hospital among the resident staff. To enter the writing competition, residents must submit a poem or essay of 1,000 words or less on the topic, "On Being a Doctor." A committee of seven was selected from Houston Methodist Hospital Education Institute to establish the judging criteria and select the winning entries. The following is the third-place winning entry for 2019.


Cholinergic stimulation of salivary secretion studied with M1 and M3 muscarinic receptor single- and double-knockout mice.
Identification of the specific muscarinic acetylcholine receptor (mAChR) subtypes mediating stimulation of salivary secretion is of considerable clinical interest. Recent pharmacological and molecular genetic studies have yielded somewhat confusing and partially contradictory results regarding the involvement of individual mAChRs in this activity. In the present study, we re-examined the roles of M(1) and M(3) mAChRs in muscarinic agonist-mediated stimulation of salivary secretion by using M(1) and M(3) receptor single-knockout (KO) mice and newly generated M(1)/M(3) receptor double-KO mice. When applied at a low dose (1 mg/kg, s.c.), the muscarinic agonist pilocarpine showed significantly reduced secretory activity in both M(1) and M(3) receptor single-KO mice. However, when applied at higher doses, pilocarpine induced only modestly reduced (5 mg/kg, s.c.) or unchanged (15 mg/kg, s.c.) salivation responses, respectively, in M(1) and M(3) receptor single-KO mice, indicating that the presence of either M(1) or M(3) receptors is sufficient to mediate robust salivary output. Quantitative reverse transcriptase-polymerase chain reaction studies with salivary gland tissue showed that the inactivation of the M(1) or M(3) mAChR genes did not lead to significantly altered mRNA levels of the remaining mAChR subtypes. Strikingly, the sialagogue activity of pilocarpine was abolished in M(1)/M(3) receptor double-KO mice. However, salivary glands from M(1)/M(3) receptor double-KO mice remained responsive to stimulation by the beta-adrenergic receptor agonist, (S)-isoproterenol. Taken together these studies support the concept that a mixture of M(1) and M(3) receptors mediates cholinergic stimulation of salivary flow.


The Arabidopsis Athb-10 (GLABRA2) is an HD-Zip protein required for regulation of root hair development.
Homeodomain-leucine zipper (HD-Zip) proteins are putative transcription factors identified only in plants. Related Arabidopsis homeobox genes, isolated by virtue of sequence conservation within the helix-3 region of the homeodomain, fall into four families based on sequence similarity. This paper reports the characterization of Athb-10, a 747 amino acid protein belonging to the fourth HD-ZIP family. The studies indicate that, although less conserved, the leucine zipper of Athb-10 can functionally replace that of Athb-2 in an in vitro DNA-binding assay. Gene mapping experiments and sequence comparison analysis revealed that Athb-10 corresponds to GLABRA2, a homeodomain protein involved in trichome development. The mRNA expression analysis revealed that Athb-10/GLABRA2 is expressed not only in trichome-bearing organs, but also in the root. The analysis of wild-type and mutant plants showed that the Athb-10/GLABRA2 gene expression in the aerial part of the plant and in the root is affected by mutations at the TTG locus. Morphological analysis of the g/2-1 mutant revealed that the gene is necessary not only for local outgrowth of the trichome, but also for the regulation of root hair development in a subset of epidermal cells. Interestingly, the development of root hair cells in a position normally occupied by non-hair cells is dependent upon the ethylene regime in which the gl2-1 plants are grown. Sequence analysis of the gl2-1 allele revealed that the mutant gene encodes a truncated protein that might still retain a partial activity responsible for the formation of aborted trichomes and for the ethylene-dependent regulation of root hair formation.


WE-G-BRB-09: CyberKnife Patient Specific QA Using a 4D Cylindrical Diode Array System.
To examine and facilitate the feasibility of the ArcCheck cylindrical diode array system as a patient specific QA device for CyberKnife radiosurgery delivery. There is an obvious necessity for CyberKnife robotic radiosurgery patient QA procedures for hypofractionated treatment of larger planned treatment volumes (PTV), e.g. prostate. This need will increase when the future CyberKnife MLC is introduced. The small unflattened CyberKnife fields, along with the variation of beam-to-detector spatial angles, pose a significant detection challenge for dosimetric systems. The feasibility of the ArcCheck (Sun Nuclear Inc.) cylindrical diode array system for patient-specific QA on the CyberKnife is demonstrated using a beam-to-diode specific angular correction that was developed and has been applied. For localization and tracking, four gold seed fiducial markers were embedded in the system's central plug. We used a Monte Carlo 1% uncertainty for the dose calculation. By disabling the Linac based corrections and applying the custom CyberKnife correction that we developed, the passing rate increased from 39.6% to 99.8% using a 3%3mm gamma criteria for a given lung case. An additional lung case passed 98.5%. In both cases, a 10% dose threshold was used. In addition, brain, trigeminal nerve and lung cases with synchrony tracking are being investigated. We demonstrated the ArcCheck feasibility for CyberKnife patient specific QA performance. The custom CK angular correction that we developed and applied showed a high passing rate for the lung cases. A verification of the polar angle response should be conducted, in addition to the azimuthal angle that was verified for Linacs. Any data that is being retrieved is additional data to the current chamber point measurement procedures.


Radon awareness and mitigation in Vermont: a public health survey.
Radon exposure is associated with an increased incidence of lung cancer, and elevated levels may be found in as many as 1 out of 15 homes. The U.S. EPA recommends testing homes for radon and mitigating over the advisory level of 4 picocuries per liter (4 pCi L(-1), or 148 Bq m(-3)). A sample population from a list of Vermont residents who had tested their residence for radon through the Vermont Department of Health and who had elevated levels were mailed a survey to assess demographic characteristics, knowledge about radon, mitigation rates, types of mitigation, as well as barriers to mitigation. The response rate was 63%. Forty-three percent of respondents mitigated. Roughly half were not completely knowledgeable of radon based upon the ability to associate radon exposure with lung cancer risk. Reasons not to mitigate radon levels in homes were cost and lack of concern over elevated levels. A multivariate logistic regression analysis revealed factors associated with mitigating: an education level of college or higher (p = 0.02), concern that a high radon level would affect real estate value (p = 0.04), and home age less than 10 y (p = 0.05). In summary, less than half of Vermonters with elevated radon levels participating in the Department of Health program mitigated. We identify factors associated with radon mitigation that may lead to improved radon education and mitigation practice.


Difference between human red blood cell Na+-Li+ countertransport and renal Na+-H+ exchange.
Several laboratories have reported that the activities of sodium-lithium countertransport are increased in red blood cells from patients with essential hypertension. Based on the many similarities between this transport system and the renal sodium-proton exchanger, a hypothesis has been put forth in the literature that increased red blood cell sodium-lithium countertransport activity may be a marker for increased sodium-proton exchange activity in the renal proximal tubule. The present studies were designed to test the hypothesis that sodium-lithium countertransport in red blood cells from humans or rabbits is mediated by the same transport mechanism that mediates sodium-proton exchange in the renal brush border from those species. Similar to what has been reported for the rabbit, the present studies show that an amiloride-sensitive sodium-proton exchanger is present in human renal brush border vesicles. However, Na+-Li+ countertransport in human and rabbit red blood cells, assayed under several different conditions, was not inhibited by amiloride. In agreement with what has been reported for humans, the present studies show that extracellular proton-stimulated sodium efflux is inhibited by amiloride in rabbit red blood cells. These data demonstrate a difference (amiloride sensitivity) between the red blood cell sodium-lithium countertransporter and the renal brush border sodium-proton exchanger in humans and rabbits. These experiments detract from the hypothesis that increased red blood cell sodium-lithium countertransport activity in patients with essential hypertension is a marker for increased sodium-proton exchange activity in the renal brush border.


Methodology and measurement of the effectiveness of Haemophilus influenzae type b vaccine: systematic review.
The use of the highly effective Haemophilus influenzae type b (Hib) conjugate vaccine has increased globally. We review the benefits and limitations of studies measuring Hib vaccine effectiveness (VE). We critically examine the case-control approach by assessing the similarities and differences in methodology and findings and discuss the need for future Hib VE studies. In the absence of good surveillance data, vaccine effectiveness studies can play an important role, particularly with the increasing use of pneumococcal vaccine that has not been well tested under field conditions in less developed countries. However, the effectiveness of Hib vaccine has been well documented so the need for future VE Hib studies is minimal.


Systolic dysfunction of systemic ventricle in patients who underwent a Fontan operation.
Öner T, Ballı Ş, Hekim Yılmaz E, Bulut MO, Şaşmazel A, Çelebi A. Systolic dysfunction of systemic ventricle in patients who underwent a Fontan operation. Turk J Pediatr 2019; 61: 925-930. The aim of the study was to report the clinical and echocardiographic data of patients who underwent a Fontan operation, and define the group with systolic dysfunction in the systemic ventricle observed during postoperative follow-up. The medical records of 183 patients [mean age: 10.93 ± 5.89 years (range: 2.5-45 years)] who were referred to our center and underwent a Fontan operation were retrospectively reviewed. The clinical, echocardiographic, and postoperative follow-up data of the patients were recorded. Preoperatively, 68 (37.2%) patients experienced pulmonary stenosis, while 41 (22.4%) had pulmonary atresia and 74 (40.4%) had pulmonary hypertension. The most common pathology in patients who were ineligible for biventricular repair was tricuspid atresia, seen in 51 patients (27.9%), followed by double-inlet left ventricle pathologies in 40 patients (21.9%). In total, 38 (20.7%) patients had a biventricular structure; among those with a single ventricular structure, the systemic ventricle involved was the right ventricle in 51 (27.9%) patients and the left ventricle in 94 (51.4%) patients. During follow-up, a total of 31 (16.9%) patients underwent catheterization and ventricular systolic functions were preserved in 168 (91.8%) patients. In Fifteen (8.2%) patients developed systolic dysfunction (ejection fraction < 50%). The mean age of the group developing systolic dysfunction was 15.6 ± 2.63 years (median, 13 years; range: 5-45 years). Of this group, 10 patients had a left ventricular structure of the systemic ventricle, 14 had atrial situs solitus, and 4 had moderate atrioventricular valve insufficiency. Even if the systemic ventricle is in the left ventricular structure, systolic dysfunction in the systemic ventricle develops, especially after the first 10 years, and this makes us think rudimentary ventricle function should also be carefully monitored for intra-univentricular diastolic asynchrony.


Spectral focusing dual-comb coherent anti-Stokes Raman spectroscopic imaging.
Coherent Raman microscopy provides label-free imaging by interrogating the intrinsic vibration of molecules. Here, we present a concept in a 100 MHz dual-comb scheme to facilitate high-speed and broadband coherent anti-Stokes Raman spectroscopic imaging based on down-converted, automatically varying delay-time in spectral focusing excitation. A rapid measurement of vibrational microspectroscopy on submicrosecond scale over a spectral span ∼700 cm-1 with a refresh rate of 1.2 kHz provides access to well-resolved molecular signatures within the fingerprint region. We demonstrate hyperspectral images of a mixture of spatially inhomogeneous distributions of chemical substances.


Gastrointestinal release behaviour of modified-release drug products: dynamic dissolution testing of mesalazine formulations.
The aminosalicylate mesalazine (mesalamine) forms the mainstay of treatment in ulcerative colitis (UC), a disease for which many commercial modified-release products have been developed with the aim of providing targeted gastrointestinal release. The release profiles of five of these commercial formulations were evaluated in bicarbonate buffer using a novel dissolution model that mimics the dynamic conditions of the gastrointestinal tract. Monolithic and multi-particulate mesalazine formulations with pH-dependent and/or independent release mechanisms were evaluated (Asacol(®) 800, Octasa(®), Mezavant(®) XL, Salofalk(®), Pentasa(®)), and each of the products displayed a distinctive dissolution profile. The dissolution results for Mezavant(®) XL (Lialda(®)) (lag time 290 min) demonstrated good correlation with previously reported in vivo disintegration times assessed by gamma-scintigraphy in humans. Octasa(®) showed a similar lag time to Mezavant(®) XL. Drug release from Asacol(®) 800 (Asacol(®) HD) showed a wide standard deviation, reflecting the great variability in vivo. Salofalk(®) displayed both delayed release and extended release characteristics. Pentasa(®) released more than 50% of its drug load in the stomach compartment of the model, which is attributed to the absence of a gastro-resistant coating in this product. The new dissolution method provided a realistic and discriminative in vitro assessment of mesalazine release from different formulations. These results demonstrate that this strategy can be used to predict intestinal release behaviour, and potentially aid the rational design of products developed to target different sites of the gut.


The role of executive functions in the pragmatic skills of children age 4-5.
Several studies suggest that pragmatic skills (PS) (i.e., social communication) deficits may be linked to executive dysfunction (i.e., cognitive processes required for the regulation of new and complex behaviors) in patients with frontal brain injuries. If impairment of executive functions (EF) causes PS deficits in otherwise healthy adults, could this mean that EF are necessary for the normal functioning of PS, even more so than cognitive maturation? If so, children with highly developed EF should exhibit higher levels of PS. This study aimed to examine the link between EF and PS among normally developing children. A secondary goal was to compare this relationship to that between intellectual quotient (IQ) and PS in order to determine which predictor explained the most variance. Participants were 70 French-speaking preschool children (3;10-5;7 years old). The PS coding system, an observational tool developed for this study, was used to codify the children's PS during a semi-structured conversation with a research assistant. Five types of EF processes were evaluated: self-control, inhibition, flexibility, working memory and planning. IQ was estimated by tallying the scores on a receptive vocabulary test and a visuoconstructive abilities test. The results of the test of differences between correlation coefficients suggest that EF contributed significantly more than IQ to the PS exhibited by preschoolers during conversation. More specifically, higher inhibition skills were correlated with a decrease in talkativeness and assertiveness. EF also appeared to foster quality of speech by promoting the ability to produce fluid utterances, free of unnecessary repetition or hesitation. Moreover, children with a high working memory capacity were more likely to formulate contingent answers and produce utterances that could be clearly understood by the interlocutor. Overall, these findings help us better understand how EF may assist children in everyday social interactions.


A time dependent kinetic small angle neutron scattering study of a novel YFe phase.
Crystallization of amorphous Y67Fe33 into the YFe2 C15 Laves phase via a novel 'YFe' intermediate phase has been observed through to completion using time-resolved small angle neutron scattering (SANS). The nucleation and growth kinetics of the phase transformations have been studied at annealing temperatures below the crystallization temperatures for both the 'YFe' phase and the YFe2 phase. The SANS results agree with previously reported neutron diffraction and SANS data. At the annealing temperatures of 360, 370 and 380 °C, changes in the scattering intensity I(Q) occur as a result of the contrast between the amorphous matrix and the nucleating and growing Y and 'YFe' phases. Critical scattering occurs during each of the isotherms, relating to the full crystallization of Y67Fe33, and extrapolation gives a crystallization temperature of 382 °C. Beyond critical scattering, isotherms at 435, 450, and 465 °C reveal the details of the continuing transformation of the 'YFe' intermediate phase into the YFe2 C15 Laves phase.


Borondifluoride complexes of hemicurcuminoids as bio-inspired push-pull dyes for bioimaging.
Hemicurcuminoids are based on half of the π-conjugated backbone of curcuminoids. The synthesis of a series of such systems and their borondifluoride complexes is described. The electrochemical and photophysical properties of difluorodioxaborine species were investigated as a function of the nature of electron donor and acceptor groups appended at either terminal positions of the molecular backbone. The emissive character of these dipolar dyes was attributed to an intraligand charge transfer process, leading to fluorescence emission that is strongly dependent on solvent polarity. Quasi-quantitative quenching of fluorescence in high polarity solvents was attributed to photoinduced electron transfer. These dyes were shown to behave as versatile fluorophores. Indeed, they display efficient two-photon excited fluorescence emission leading to high two-photon brightness values. Furthermore, they form nanoparticles in water whose fluorescence emission quantum yield is less than that of the dye in solution, owing to aggregation-induced fluorescence quenching. When cos7 living cells were exposed to these weakly-emitting nanoparticles, one- and two-photon excited fluorescence spectra showed a strong emission within the cytoplasm that originated from the individual molecules. Dye uptake thus involved a disaggregation mechanism at the cell membrane which restored fluorescence emission. This off-on fluorescence switching allows a selective optical monitoring of those molecules that do enter the cell, which offers improved sensitivity and selectivity of detection for bioimaging purposes.


Lateral patch-clamping in a standard 1536-well microplate format.
Lateral patch-clamping has emerged as a chip-based platform for automation of the conventional patch-clamp technique, the 'gold' standard for studying cellular ion channels. The conventional technique, as it relies on skilled-maneuver of glass micropipettes to patch cells, is extremely delicate, low in throughput, and thus cannot be used for primary screening of compounds against ion channels. Direct integration of glass capillaries on silicon provides lateral junctions for automated trapping and patching of cells. We demonstrate here a method of scaling up the lateral junctions to a standard 1536-well microtiter plate format. A single unit of 1536-well plate has been formed here on a 9 mm by 9 mm microstructured silicon with the inclusive of 16 wells molded in a capping layer made of polydimethylsiloxane (PDMS). The silicon substrate provides integrated glass capillaries (total 12) and their associated microfluidic network. Each glass capillary has an independent access through a dedicated well in PDMS and leads to a centralized channel in which cell suspension can be delivered through one of the remaining 4 wells. The unit has been tested on RBL-1 cells by recording whole-cell activity from inwardly rectifying endogenous potassium channels. A revised test protocol has been prescribed to avoid inaccurate readings due to altered ionic composition of the recording buffer when a typical suction is applied to capture cells.


Adaptive Evolution of Phosphorus Metabolism in Prochlorococcus.
Inorganic phosphorus is scarce in the eastern Mediterranean Sea, where the high-light-adapted ecotype HLI of the marine picocyanobacterium Prochlorococcus marinus thrives. Physiological and regulatory control of phosphorus acquisition and partitioning has been observed in HLI both in culture and in the field; however, the optimization of phosphorus metabolism and associated gains for its phosphorus-limited-growth (PLG) phenotype have not been studied. Here, we reconstructed a genome-scale metabolic network of the HLI axenic strain MED4 (iJC568), consisting of 568 metabolic genes in relation to 794 reactions involving 680 metabolites distributed in 6 subcellular locations. iJC568 was used to quantify metabolic fluxes under PLG conditions, and we observed a close correspondence between experimental and computed fluxes. We found that MED4 has minimized its dependence on intracellular phosphate, not only through drastic depletion of phosphorus-containing biomass components but also through network-wide reductions in phosphate-reaction participation and the loss of a key enzyme, succinate dehydrogenase. These alterations occur despite the stringency of having relatively few pathway redundancies and an extremely high proportion of essential metabolic genes (47%; defined as the percentage of lethal in silico gene knockouts). These strategies are examples of nutrient-controlled adaptive evolution and confer a dramatic growth rate advantage to MED4 in phosphorus-limited regions. IMPORTANCE Microbes are known to employ three basic strategies to compete for limiting elemental resources: (i) cell quotas may be adjusted by alterations to cell physiology or by substitution of a more plentiful resource, (ii) stressed cells may synthesize high-affinity transporters, and (iii) cells may access more costly sources from internal stores, by degradation, or by petitioning other microbes. In the case of phosphorus, a limiting resource in vast oceanic regions, the cosmopolitan cyanobacterium Prochlorococcus marinus thrives by adopting all three strategies and a fourth, previously unknown strategy. By generating a detailed model of its metabolism, we found that strain MED4 has evolved a way to reduce its dependence on phosphate by minimizing the number of enzymes involved in phosphate transformations, despite the stringency of nearly half of its metabolic genes being essential for survival. Relieving phosphorus limitation, both physiologically and throughout intermediate metabolism, substantially improves phosphorus-specific growth rates.


Identification of DNA-binding proteins by combining auto-cross covariance transformation and ensemble learning.
DNA-binding proteins play a pivotal role in various intra- and extra-cellular activities ranging from DNA replication to gene expression control. With the rapid development of next generation of sequencing technique, the number of protein sequences is unprecedentedly increasing. Thus it is necessary to develop computational methods to identify the DNA-binding proteins only based on the protein sequence information. In this study, a novel method called iDNA-KACC is presented, which combines the Support Vector Machine (SVM) and the auto-cross covariance transformation. The protein sequences are first converted into profile-based protein representation, and then converted into a series of fixed-length vectors by the auto-cross covariance transformation with Kmer composition. The sequence order effect can be effectively captured by this scheme. These vectors are then fed into Support Vector Machine (SVM) to discriminate the DNA-binding proteins from the non DNA-binding ones. iDNA-KACC achieves an overall accuracy of 75.16% and Matthew correlation coefficient of 0.5 by a rigorous jackknife test. Its performance is further improved by employing an ensemble learning approach, and the improved predictor is called iDNA-KACC-EL. Experimental results on an independent dataset shows that iDNA-KACC-EL outperforms all the other state-of-the-art predictors, indicating that it would be a useful computational tool for DNA binding protein identification. .


WE-C-217BCD-02: Design of an MR Compatible Rotating Anode X-Ray Tube.
To design a rotating anode X-ray tube capable of operating in strong magnetic field environments. This tube design can be used in 'close proximity" hybrid X-ray/MR system geometries where the imaging fields of view are separated by only ∼1.2 meters. Existing rotating anode X-ray tube designs fail in strong magnetic field environments because the fields alter the electron trajectories in the tube and act as a brake on the induction motor, reducing the rotation speed of the anode. We propose an X- ray tube design that utilizes optimized resistive coils to shield a fraction of the MR fringe field. The remainder of the correction is performed using bias voltages on electrodes adjacent to the x-ray tube filament. Furthermore, we replace the induction motor with a novel motor design that is analogous to a three-phase brushed DC motor with the MR fringe field serving as the stator field. Space charge simulations of the electron optics show that the combined magnetostatic/electrostatic method can correct for a magnetic field strength of 152 mT with approximately 590 A/cm2 applied to the shielding coils and a 35 kV potential difference applied to the bias electrodes. A prototype of the motor design was machined and assembled. The performance of this prototype motor was evaluated at various magnetic field strengths, and was found to accelerate to the minimum operating speed of 3000 rpm in 10 seconds for an external field of 60 mT. The space charge simulations validate that the electron trajectories can be controlled using our combined approach. Testing of the motor prototype demonstrates that our design outperforms existing induction motors in strong magnetic field environments. Integrating this design with our modified flat panel detector will allow, for the first time, a "close proximity" hybrid system in which imaging performance is not compromised. NIH R01 EB007626 Richard M. Lucas Foundation Stanford Bio-X Fellowship.


Competitive action video game players display rightward error bias during on-line video game play.
Research in asymmetrical visuospatial attention has identified a leftward bias in the general population across a variety of measures including visual attention and line-bisection tasks. In addition, increases in rightward collisions, or bumping, during visuospatial navigation tasks have been demonstrated in real world and virtual environments. However, little research has investigated these biases beyond the laboratory. The present study uses a semi-naturalistic approach and the online video game streaming service Twitch to examine navigational errors and assaults as skilled action video game players (n = 60) compete in Counter Strike: Global Offensive. This study showed a significant rightward bias in both fatal assaults and navigational errors. Analysis using the in-game ranking system as a measure of skill failed to show a relationship between bias and skill. These results suggest that a leftward visuospatial bias may exist in skilled players during online video game play. However, the present study was unable to account for some factors such as environmental symmetry and player handedness. In conclusion, video game streaming is a promising method for behavioural research in the future, however further study is required before one can determine whether these results are an artefact of the method applied, or representative of a genuine rightward bias.


Circadian rhythm of salivary serotonin in patients with major depressive disorder.
The present study aimed to explore the circadian rhythm of salivary serotonin in patients with major depressive disorder before and after treatment with fluoxetine and its relationship with clinical therapeutic effect. This study investigated salivary serotonin in 13 outpatients with major depressive disorder and age- and sex-matched healthy controls. Depressed patients received six weeks fluoxetine treatment (20 mg/day), saliva was collected before and four weeks after treatment. A total of 8 time-point salivary serotonin was measured across the whole day. Multi-oscillator cosinor model was used to fit the rhythms. Serotonin concentration in saliva ranged from 0.32 ng/ml to 9.62 ng/ml. Salivary serotonin showed prominent circadian rhythm in 91% depressed patients and 92% healthy subjects. Circadian amplitude tend to be higher after fluoxetine treatment in depressed patients, so as the ultradian cycle amplitude. The serotonin circadian amplitude (After minus Before) was positively correlated with the decrease of Zung Self-Rating Depression Scale(SDS) scores at day 42 whereas there was no such correlation at day 28. There was no significant difference in the parameters of mesor, acrophase, harmonic and area under curve among three groups. Salivary serotonin in patients with major depressive disorder showed clear circadian rhythm. The relationship between the increase of salivary serotonin amplitude and clinical response deserve further study.


High-speed graphene transistors with a self-aligned nanowire gate.
Graphene has attracted considerable interest as a potential new electronic material. With its high carrier mobility, graphene is of particular interest for ultrahigh-speed radio-frequency electronics. However, conventional device fabrication processes cannot readily be applied to produce high-speed graphene transistors because they often introduce significant defects into the monolayer of carbon lattices and severely degrade the device performance. Here we report an approach to the fabrication of high-speed graphene transistors with a self-aligned nanowire gate to prevent such degradation. A Co(2)Si-Al(2)O(3) core-shell nanowire is used as the gate, with the source and drain electrodes defined through a self-alignment process and the channel length defined by the nanowire diameter. The physical assembly of the nanowire gate preserves the high carrier mobility in graphene, and the self-alignment process ensures that the edges of the source, drain and gate electrodes are automatically and precisely positioned so that no overlapping or significant gaps exist between these electrodes, thus minimizing access resistance. It therefore allows for transistor performance not previously possible. Graphene transistors with a channel length as low as 140 nm have been fabricated with the highest scaled on-current (3.32 mA μm(-1)) and transconductance (1.27 mS μm(-1)) reported so far. Significantly, on-chip microwave measurements demonstrate that the self-aligned devices have a high intrinsic cut-off (transit) frequency of f(T) = 100-300 GHz, with the extrinsic f(T) (in the range of a few gigahertz) largely limited by parasitic pad capacitance. The reported intrinsic f(T) of the graphene transistors is comparable to that of the very best high-electron-mobility transistors with similar gate lengths.


Tracing path-guided apparent motion in human primary visual cortex V1.
Vision is a constructive process. For example, a square, flashed at two distinct locations one after the other, appears to move smoothly between the two locations rather than as two separate flashes (apparent motion). Apparent motion is usually perceived along the shortest path between locations. Previous studies have shown that retinotopic activity in V1 correlates well with the subjective filling-in in apparent motion. If V1 activity truly reflects illusory motion, it should flexibly reflect filling-in of any path, subjectively perceived. Here, we used a path-guided apparent motion paradigm in which a faint cue, presented in addition to the squares, leads to a curved illusory motion path. We found retinotopic activity in V1 to reflect the illusory filling-in of the curved path, similarly to filling-in with linear, shortest paths. Moreover, our results show that activity along the linear path was less selective to stimulus conditions than the activity along the curved path. This finding may be interpreted as V1 activity representing a small subset of infinitely many possible solutions to ambiguous stimuli, whilst giving more weight to the shortest path/energy solution.


Health-Related Resource-Use Measurement Instruments for Intersectoral Costs and Benefits in the Education and Criminal Justice Sectors.
Intersectoral costs and benefits (ICBs), i.e. costs and benefits of healthcare interventions outside the healthcare sector, can be a crucial component in economic evaluations from the societal perspective. Pivotal to their estimation is the existence of sound resource-use measurement (RUM) instruments; however, RUM instruments for ICBs in the education or criminal justice sectors have not yet been systematically collated or their psychometric quality assessed. This review aims to fill this gap. To identify relevant instruments, the Database of Instruments for Resource Use Measurement (DIRUM) was searched. Additionally, a systematic literature review was conducted in seven electronic databases to detect instruments containing ICB items used in economic evaluations. Finally, studies evaluating the psychometric quality of these instruments were searched. Twenty-six unique instruments were included. Most frequently, ICB items measured school absenteeism, tutoring, classroom assistance or contacts with legal representatives, police custody/prison detainment and court appearances, with the highest number of items listed in the Client Service Receipt Inventory/Client Sociodemographic and Service Receipt Inventory/Client Service Receipt Inventory-Children's Version (CSRI/CSSRI/CSRI-C), Studying the Scope of Parental Expenditures (SCOPE) and Self-Harm Intervention, Family Therapy (SHIFT) instruments. ICBs in the education sector were especially relevant for age-related developmental disorders and chronic diseases, while criminal justice resource use seems more important in mental health, including alcohol-related disorders or substance abuse. Evidence on the validity or reliability of ICB items was published for two instruments only. With a heterogeneous variety of ICBs found to be relevant for several disease areas but many ICB instruments applied in one study only (21/26 instruments), setting-up an international task force to, for example, develop an internationally adaptable instrument is recommended.


Activation and functional significance of the renin-angiotensin system in mice with cardiac restricted overexpression of tumor necrosis factor.
The functional significance of cross-regulation between the renin-angiotensin system (RAS) and tumor necrosis factor (TNF) has been established in nonmyocyte cell types; however, the degree and functional significance of the interaction between RAS and TNF has not been characterized in the heart. We examined the expression of components of the RAS in a line of transgenic mice (MHCsTNF) with cardiac restricted overexpression of TNF. When examined at 4, 8, and 12 weeks of age, the MHCsTNF mice had increased activation of myocardial RAS, as shown by an increase in ACE mRNA level and ACE activity and increased angiotensin II peptide levels. Furthermore, myocardial angiotensin receptor mRNA and protein levels were reduced in the MHCsTNF mice, consistent with homologous desensitization of the receptors. However, expression of renin and angiotensinogen was not increased in MHCsTNF mice compared with littermate controls. To determine the functional significance of RAS activation in the MHCsTNF mice, we treated the mice with an angiotensin type I receptor antagonist, losartan (30 mg/kg), or diluent from 4 to 8 weeks of age. Analysis of cardiac structure with MRI showed that treatment with losartan normalized left ventricular mass and wall thickness. Furthermore, treatment with losartan reduced myocardial collagen content and reduced the incidence of myocyte apoptosis. Taken together, these results show that there are functionally significant interactions between RAS and TNF in the heart and that these interactions play an important role in the development and progression of left ventricular remodeling.


Motor cortical processing is causally involved in object recognition.
Motor activity during vicarious experience of actions is a widely reported and studied phenomenon, and motor system activity also accompanies observation of graspable objects in the absence of any actions. Such motor activity is thought to reflect simulation of the observed action, or preparation to interact with the object, respectively. Here, in an initial exploratory study, we ask whether motor activity during observation of object directed actions is involved in processes related to recognition of the object after initial exposure. Single pulse Transcranial Magnetic Stimulation (TMS) was applied over the thumb representation of the motor cortex, or over the vertex, during observation of a model thumb typing on a cell-phone, and performance on a phone recognition task at the end of the trial was assessed. Disrupting motor processing over the thumb representation 100 ms after the onset of the typing video impaired the ability to recognize the phone in the recognition test, whereas there was no such effect for TMS applied over the vertex and no TMS trials. Furthermore, this effect only manifested for videos observed from the first person perspective. In an additional control condition, there was no evidence for any effects of TMS to the thumb representation or vertex when observing and recognizing non-action related shape stimuli. Overall, these data provide evidence that motor cortical processing during observation of object-directed actions from a first person perspective is causally linked to the formation of enduring representations of objects-of-action.


Base sequence dependence of in vitro translesional DNA replication past a bulky lesion catalyzed by the exo- Klenow fragment of Pol I.
The effects of base sequence, specifically different pyrimidines flanking a bulky DNA adduct, on translesional synthesis in vitro catalyzed by the Klenow fragment of Escherichia coli Pol I (exo(-)) was investigated. The bulky lesion was derived from the binding of a benzo[a]pyrene diol epoxide isomer [(+)-anti-BPDE] to N(2)-guanine (G*). Four different 43-base long oligonucleotide templates were constructed with G* at a site 19 bases from the 5'-end. All bases were identical, except for the pyrimidines, X or Y, flanking G* (sequence context 5'-.XGY., with X, Y = C and/or T). In all cases, the adduct G* slows primer extension beyond G* more than it slows the insertion of a dNTP opposite G* (A and G were predominantly inserted opposite G, with A > G). Depending on X or Y, full lesion bypass differed by factors of approximately 1.5-5 ( approximately 0.6-3.0% bypass efficiencies). A downstream T flanking G on the 5'-side instead of C favors full lesion bypass, while an upstream C flanking G* is more favorable than a T. Various deletion products resulting from misaligned template-primer intermediates are particularly dominant ( approximately 5.0-6.0% efficiencies) with an upstream flanking C, while a 3'-flanking T lowers the levels of deletion products ( approximately 0.5-2.5% efficiencies). The kinetics of (1) single dNTP insertion opposite G* and (2) extension of the primer beyond G* by a single dNTP, or in the presence of all four dNTPs, with different 3'-terminal primer bases (Z) opposite G* were investigated. Unusually efficient primer extension efficiencies beyond the adduct (approaching approximately 90%) was found with Z = T in the case of sequences with 3'-flanking upstream C rather than T. These effects are traced to misaligned slipped frameshift intermediates arising from the pairing of pairs of downstream template base sequences (up to 4-6 bases from G*) with the 3'-terminal primer base and its 5'-flanking base. The latter depend on the base Y and on the base preferentially inserted opposite the adduct. Thus, downstream template sequences as well as the bases flanking G* influence DNA translesion synthesis.


Impact of lipoprotein(a) levels and apolipoprotein(a) isoform size on risk of coronary heart disease.
Observational and genetic studies have shown that lipoprotein(a) [Lp(a)] levels and apolipoprotein(a) [apo(a)] isoform size are both associated with coronary heart disease (CHD) risk, but the relative independence of these risk factors remains unclear. Clarification of this uncertainty is relevant to the potential of future Lp(a)-lowering therapies for the prevention of CHD. Plasma Lp(a) levels and apo(a) isoform size, estimated by the number of kringle IV (KIV) repeats, were measured in 995 patients with CHD and 998 control subjects. The associations between CHD risk and fifths of Lp(a) levels were assessed before and after adjustment for KIV repeats and, conversely, the associations between CHD risk and fifths of KIV repeats were assessed before and after adjustment for Lp(a) levels. Individuals in the top fifth of Lp(a) levels had more than a twofold higher risk of CHD compared with those in the bottom fifth, and this association was materially unaltered after adjustment for KIV repeats [odds ratio (OR) 2.05, 95% confidence interval (CI) 1.38-3.04, P < 0.001]. Furthermore, almost all of the excess risk was restricted to the two-fifths of the population with the highest Lp(a) levels. Individuals in the bottom fifth of KIV repeats had about a twofold higher risk of CHD compared with those in the top fifth, but this association was no longer significant after adjustment for Lp(a) levels (OR 1.13, 95% CI 0.77-1.66, P = 0.94). The effect of KIV repeats on CHD risk is mediated through their impact on Lp(a) levels, suggesting that absolute levels of Lp(a), rather than apo(a) isoform size, are the main determinant of CHD risk.


Activation of KRAS Mediates Resistance to Targeted Therapy in MET Exon 14-mutant Non-small Cell Lung Cancer.
MET exon 14 splice site alterations that cause exon skipping at the mRNA level (METex14) are actionable oncogenic drivers amenable to therapy with MET tyrosine kinase inhibitors (TKI); however, secondary resistance eventually arises in most cases while other tumors display primary resistance. Beyond relatively uncommon on-target MET kinase domain mutations, mechanisms underlying primary and acquired resistance remain unclear. We examined clinical and genomic data from 113 patients with lung cancer with METex14. MET TKI resistance due to KRAS mutation was functionally evaluated using in vivo and in vitro models. Five of 113 patients (4.4%) with METex14 had concurrent KRAS G12 mutations, a rate of KRAS cooccurrence significantly higher than in other major driver-defined lung cancer subsets. In one patient, the KRAS mutation was acquired post-crizotinib, while the remaining 4 METex14 patients harbored the KRAS mutation prior to MET TKI therapy. Gene set enrichment analysis of transcriptomic data from lung cancers with METex14 revealed preferential activation of the KRAS pathway. Moreover, expression of oncogenic KRAS enhanced MET expression. Using isogenic and patient-derived models, we show that KRAS mutation results in constitutive activation of RAS/ERK signaling and resistance to MET inhibition. Dual inhibition of MET or EGFR/ERBB2 and MEK reduced growth of cell line and xenograft models. KRAS mutation is a recurrent mechanism of primary and secondary resistance to MET TKIs in METex14 lung cancers. Dual inhibition of MET or EGFR/ERBB2 and MEK may represent a potential therapeutic approach in this molecular cohort.


The role of ubiquitin ligases in the control of organ specific autoimmunity.
Diabetes mellitus is characterized by chronic hyperglycemia caused by a deficiency in insulin action, insulin secretion or both. Type 1 diabetes is classified as the destruction of beta cells leading to a deficiency in insulin production. Type1 diabetes accounts for 5-10% of patients with diabetes and most commonly is caused by the autoimmune destruction of the beta cells in the pancreas. The adaptive immune system is composed of antigen specific T and B lymphocytes which play a central role in protecting the human body from infectious pathogens but occasionally autoreactive T and B cells can escape immune tolerance, become activated and induce autoimmune diseases. Naïve T cells require two distinct signals one delivered via the antigen receptor and the second through the costimulatory receptor CD28 that leads to the induction of IL-2 gene transcription. IL-2 is an important T cell growth factor that can influence both immunity and tolerance. Given its pivotal role it is not surprising that the immune system places strict regulation over Il2 gene transcription that is controlled by a number of E3 ubiquitin ligases that modulate TCR and CD28 signaling. This review will examine how different E3 ligases function to control T effector cell differentiation and how studies in gene knockout animal models has been crucial in understanding how these proteins function in vivo to regulate immune tolerance in the peripheral circulation.


Endovascular treatment of brain arteriovenous malformations with prolonged intranidal Onyx injection technique: long-term results in 350 consecutive patients with completed endovascular treatment course.
The purpose of this study was to present the authors' clinical experience and long-term angiographic and clinical follow-up results in 350 patients with brain arteriovenous malformations (AVMs) treated using prolonged intranidal Onyx injection with a very slow "staged" reflux technique described by the authors. Three hundred and fifty consecutive patients with brain AVMs treated using Onyx between 1999 and 2008 and in whom definitive status for endovascular treatment was reached are presented. There were 206 (59%) male and 144 (41%) female patients, with a mean age of 34 years. There were 607 endovascular sessions performed. Onyx was the only agent used for intranidal injections in all patients, but in 42 patients high-concentration N-butyl cyanoacrylate glue was used adjunctively to close high-flow direct arteriovenous intra- or perinidal fistulas, or when a feeding vessel or nidus perforation and/or dissection occurred. Angiographically confirmed obliteration was achieved in 179 patients (51%) with only endovascular treatment; 1 patient died due to intracranial hemorrhage after the treatment. Twenty-two patients underwent resection, and 136 patients were sent to radiosurgery after endovascular treatment. In 4 patients embolization therapy was discontinued, and 5 additional patients refused the suggested complementary surgery. In all 178 surviving patients who had angiographically confirmed AVM obliteration by embolization alone, 1-8 years of control angiography (mean 47 months) confirmed stable obliteration, except for 2 patients in whom a very small recruitment was noted in the 1st year on control angiography studies, despite initial apparent total obliteration (recanalization rate 1.1%). In the entire series, 5 patients died; the mortality rate was 1.4%. The permanent morbidity rate was 7.1%. With the prolonged intranidal injection technique described herein, Onyx allows the practitioner to achieve higher rates of anatomical cures compared with the cure rates obtained previously with other embolic agents. More importantly, due to this technique's much more effective intranidal penetration, it allows high-grade AVMs to be made radiosurgically treatable in a group of patients for whom there has been no treatment alternative.


Visibility-dependent depth of focus for incoherent sinusoidal sources.
A visibility-dependent depth of focus is developed for incoherent sinusoidal sources. This analysis is based on a derived relationship between the visibility and the optical transfer function (OTF). The formalism developed is general for any aperture topology and arbitrary orientation of sinusoidal sources. To illustrate the application of the method the cases of an annular aperture and a Gaussian aperture are analyzed. It is found that as the level of defect of defocus increases, the maximum visibility for which a particular spatial frequency can be resolved decreases.


Hand Compartment Syndrome Due to N-acetylcysteine Extravasation.
N-acetylcysteine (NAC) is the antidote for acetaminophen (APAP)-induced hepatotoxicity. Both intravenous (IV) and oral (PO) NAC formulations are available with equal efficacy. Adverse events from either preparation are rare. We describe a hand compartment syndrome after extravasation of NAC requiring emergent fasciotomy during phase three of treatment for suspected APAP toxicity. Extravasation injuries leading to compartment syndrome are rare. It is unclear whether IV NAC induced a direct tissue-toxic insult, or functioned as a space-occupying lesion to cause a compartment syndrome. Compartment syndrome from extravasation of NAC is possible. In cases where IV access is difficult, PO NAC is an alternative.


Negative frequency-dependent selection or alternative reproductive tactics: maintenance of female polymorphism in natural populations.
Sex-limited polymorphisms have long intrigued evolutionary biologists and have been the subject of long-standing debates. The coexistence of multiple male and/or female morphs is widely believed to be maintained through negative frequency-dependent selection imposed by social interactions. However, remarkably few empirical studies have evaluated how social interactions, morph frequencies and fitness parameters relate to one another under natural conditions. Here, we test two hypotheses proposed to explain the maintenance of a female polymorphism in a species with extreme geographical variation in morph frequencies. We first elucidate how fecundity traits of the morphs vary in relation to the frequencies and densities of males and female morphs in multiple sites over multiple years. Second, we evaluate whether the two female morphs differ in resource allocation among fecundity traits, indicating alternative tactics to maximize reproductive output. We present some of the first empirical evidence collected under natural conditions that egg number and clutch mass was higher in the rarer female morph. This morph-specific fecundity advantage gradually switched with the population morph frequency. Our results further indicate that all investigated fecundity traits are negatively affected by relative male density (i.e. operational sex ratio), which confirms male harassment as selective agent. Finally, we show a clear trade-off between qualitative (egg mass) and quantitative (egg number) fecundity traits. This trade-off, however, is not morph-specific. Our reported frequency- and density-dependent fecundity patterns are consistent with the hypothesis that the polymorphism is driven by a conflict between sexes over optimal mating rate, with costly male sexual harassment driving negative frequency-dependent selection on morph fecundity.


Visualization of Water-Induced Surface Segregation of Polarons on Rutile TiO2(110).
Water-oxide surfaces are ubiquitous in nature and of widespread importance to phenomena like corrosion as well as contemporary industrial challenges such as energy production through water splitting. So far, a reasonably robust understanding of the structure of such interfaces under certain conditions has been obtained. Considerably less is known about how overlayer water modifies the inherent reactivity of oxide surfaces. Here we address this issue experimentally for rutile TiO2(110) using scanning tunneling microscopy and photoemission, with complementary density functional theory calculations. Through detailed studies of adsorbed water nanoclusters and continuous water overlayers, we determine that excess electrons in TiO2 are attracted to the top surface layer by water molecules. Measurements on methanol show similar behavior. Our results suggest that adsorbate-induced surface segregation of polarons could be a general phenomenon for technologically relevant oxide materials, with consequences for surface chemistry and the associated catalytic activity.


Antidiabetic activity of Cassia occidentalis (Linn) in normal and alloxan-induced diabetic rats.
To evaluate the hypoglycemic activity of various extracts, petroleum ether, chloroform and aqueous extract of Cassia occidentalis in normal and alloxan-induced diabetic rats. Petroleum ether, chloroform and aqueous extract of whole plant of Cassia occidentalis were orally tested at the dose of 200 mg/kg for hypoglycemic effect in normal and alloxan-induced diabetic rats. In addition, changes in body weight, serum cholesterol, triglyceride and total protein levels, assessed in the ethanol extract-treated diabetic rats, were compared with diabetic control and normal animals. Histopathological observations during 21 days treatment were also evaluated. Aqueous extract of C. occidentalis produced a significant reduction in fasting blood glucose levels in the normal and alloxan-induced diabetic rats. Apart from aqueous extract, petroleum ether extract showed activity from day 14 and chloroform extract showed activity from 7 days. Significant differences were observed in serum lipid profiles (cholesterol and triglyceride), serum protein, and changes in body weight by aqueous extract treated-diabetic animals, when compared with the diabetic control and normal animals. Concurrent histopathological studies of the pancreas of these animals showed comparable regeneration by extract which were earlier necrosed by alloxan. Aqueous extract of C. occidentalis exhibited significant antihyperglycemic activity in normal and alloxan-induced diabetic rats. They also showed improvement in parameters like body weight and serum lipid profiles as well as histopathological studies showed regeneration of β-cells of pancreas and so might be of value in diabetes treatment.


A comprehensive microscopic picture of the benzhydryl radical and cation photogeneration and interconversion through electron transfer.
Bond cleavage and bond formation are central to organic chemistry. Carbocations play a key role in our understanding of nucleophilic substitution reactions that involve both processes. The precise understanding of the mechanism and dynamics of the photogeneration of carbocations and carbon radicals is therefore an important quest. In particular, the role of electron transfer for the generation of carbocations from the radical pair is still unclear. A quantitative femtosecond absorption study is presented, with ultrabroad probing on selected donor and acceptor substituted benzhydryl chlorides irradiated with 270 nm (35 fs) pulses. The ultrafast bond cleavage within 300 fs is almost exclusively homolytic, thus leading to a radical pair. The carbocations observable in the nanosecond regime are generated from these radicals by electron transfer from the benzhydryl to the chlorine radical within the first tens of picoseconds. Their concentration is reduced by geminate recombination within hundreds of picoseconds. In moderately polar solvents this depletion almost extinguishes the cation population; in highly polar solvents free ions are still observable on the nanosecond timescale. The explanation of the experimental findings requires the microscopic realm of the intermediates to be accounted for, including their spatial and environmental distributions. The distance dependent electron transfer described by Marcus theory is combined with Smoluchowski diffusion. The depletion of the radical pair distribution at small distances causes a temporal increase of the mean distance and the observed stretched exponential electron transfer. A close accord with experiment can only be reached for a broad distribution of the nascent radical pairs. The increase in the inter-radical and inter-ion pair distance is measured directly as a shift of the UV/Vis absorption of the products. The results demonstrate that, at least for aprotic solvents, traditional descriptions of reaction mechanisms based on the concept of contact and solvent-separated pairs have to be reassessed.


Developing core sets for persons with bipolar disorder based on the International Classification of Functioning, Disability and Health.
Functioning is increasingly being taken into account when assessing the impact of bipolar disorder (BD) on the individual, as well as the effectiveness of treatments. With the International Classification of Functioning, Disability and Health (ICF), we can now rely on a globally agreed-upon framework and system for classifying the typical spectrum of problems in functioning, given the environmental context in which subjects live. ICF Core Sets are subgroups of ICF items selected to capture those aspects of functioning that are most likely to be affected by specific disorders. Within a given disorder, both Brief and Comprehensive Core Sets can be established to serve specific purposes. The aim of this paper is to outline the development process of the ICF Core Sets for BD. The final definition of ICF Core Sets for BD will be determined at an ICF Core-Sets Consensus Conference, which will integrate evidence from preliminary studies, namely (a) semi-structured interviews with people with BD in different countries, (b) a Delphi exercise with international experts participating and (c) a cross-sectional study. ICF Core Sets are being designed with the goal of providing useful standards for research, clinical practice and teaching. We believe that these ICF Core Sets for BD will stimulate research leading to improved understanding of functioning, disability and health in BD. Such research, we hope, will lead to interventions and accommodations to improve restoration and maintenance of functioning and minimise disability among people with BD throughout the world.


Avidity characterization of genetically engineered T-cells with novel and established approaches.
Adoptive transfer of genetically engineered autologous T-cells is becoming a successful therapy for cancer. The avidity of the engineered T-cells is of crucial importance for therapy success. We have in the past cloned a T-cell receptor (TCR) that recognizes an HLA-A2 (MHC class I)-restricted peptide from the prostate and breast cancer- associated antigen TARP. Herein we perform a side-by-side comparison of the TARP-specific TCR (TARP-TCR) with a newly cloned TCR specific for an HLA-A2-restricted peptide from the cytomegalovirus (CMV) pp65 antigen. Both CD8(+) T-cells and CD4(+) T-cells transduced with the HLA-A2-restricted TARP-TCR could readily be detected by multimer analysis, indicating that the binding is rather strong, since binding occured also without the CD8 co-receptor of HLA-A2. Not surprisingly, the TARP-TCR, which is directed against a self-antigen, had weaker binding to the HLA-A2/peptide complex than the CMV pp65-specific TCR (pp65-TCR), which is directed against a viral epitope. Higher peptide concentrations were needed to achieve efficient cytokine release and killing of target cells when the TARP-TCR was used. We further introduce the LigandTracer technology to study cell-cell interactions in real time by evaluating the interaction between TCR-engineered T-cells and peptide-pulsed cancer cells. We were able to successfully detect TCR-engineered T-cell binding kinetics to the target cells. We also used the xCELLigence technology to analyzed cell growth of target cells to assess the killing potency of the TCR-engineered T-cells. T-cells transduced with the pp65 - TCR exhibited more pronounced cytotoxicity, being able to kill their targets at both lower effector to target ratios and lower peptide concentrations. The combination of binding assay with functional assays yields data suggesting that TARP-TCR-engineered T-cells bind to their target, but need more antigen stimulation compared to the pp65-TCR to achieve full effector response. Nonetheless, we believe that the TARP-TCR is an attractive candidate for immunotherapy development for prostate and/or breast cancer.


A cereal-based evening meal rich in indigestible carbohydrates increases plasma butyrate the next morning.
Epidemiological studies have shown an inverse relation between a whole grain consumption and risk of type-2 diabetes and cardiovascular disease. One tentative mechanism relates to colonic metabolism of indigestible carbohydrates. In a previous study, we reported a positive relation between colonic fermentation and improved glucose tolerance. This work can be seen as an extension of that study, focusing on the tentative role of specific colonic metabolites, i.e. SCFA. Plasma concentrations of acetate, propionate, and butyrate were determined in the morning in healthy participants (5 women and 10 men, mean ± SD: 25.9 ± 3.2 y, BMI < 25) following 8 different cereal-based evening meals (50 g available starch) varying in content of indigestible carbohydrates. Each participant consumed all test meals in a random order on separate evenings. At a standardized breakfast following evening test meals, the postprandial glucose response (incremental area under the curve, 0-120 min) was inversely related to plasma butyrate (r = -0.26; P < 0.01) and acetate (r = -0.20; P < 0.05) concentrations. Evening meals composed of high-amylose barley kernels or high-β-glucan barley kernels resulted in higher plasma butyrate concentrations the following morning compared with an evening meal with white wheat bread (P < 0.05). The results support the view that cereal products rich in indigestible carbohydrates may improve glucose tolerance through a mechanism involving colonic fermentation and generation of SCFA, where in particular butyric acid may be involved. This mechanism may be one explanation by which whole grain is protective against type 2 diabetes and cardiovascular disease.


Genomic changes following the reversal of a Y chromosome to an autosome in Drosophila pseudoobscura.
Robertsonian translocations resulting in fusions between sex chromosomes and autosomes shape karyotype evolution by creating new sex chromosomes from autosomes. These translocations can also reverse sex chromosomes back into autosomes, which is especially intriguing given the dramatic differences between autosomes and sex chromosomes. To study the genomic events following a Y chromosome reversal, we investigated an autosome-Y translocation in Drosophila pseudoobscura. The ancestral Y chromosome fused to a small autosome (the dot chromosome) approximately 10-15 Mya. We used single molecule real-time sequencing reads to assemble the D. pseudoobscura dot chromosome, including this Y-to-dot translocation. We find that the intervening sequence between the ancestral Y and the rest of the dot chromosome is only ∼78 Kb and is not repeat-dense, suggesting that the centromere now falls outside, rather than between, the fused chromosomes. The Y-to-dot region is 100 times smaller than the D. melanogaster Y chromosome, owing to changes in repeat landscape. However, we do not find a consistent reduction in intron sizes across the Y-to-dot region. Instead, deletions in intergenic regions and possibly a small ancestral Y chromosome size may explain the compact size of the Y-to-dot translocation.


Identification of a common molecular pathway in hypertensive renal damage: comparison of rat and human gene expression profiles.
There is a common structural progression in hypertensive renal damage with early arterial damage and fibrosis in the juxtamedullary cortex. The present investigation identifies a common pathway using three-gene expression profiles from hypertensive rat models: 60-week-old spontaneously hypertensive rat (SHR), salt-loaded stroke-prone SHR (SHRSP), and the non-clipped kidney after 24 weeks of two-kidney, one-clip hypertension (2K1C). Kidney damage was scored using a specialized system. Gene-expression profiles were determined using microarrays and validated using a panel of 47 genes by quantitative real-time PCR. All groups showed kidney damage (SHRs: 0.32 ± 0.09 vs. Wistar-Kyoto rats: 0.06 ± 0.03; 2K1C: 0.27 ± 0.13 vs. pooled controls: 0.01 ± 0.01; SHRSP: 1.13 ± 0.14 vs. WKY: 0.04 ± 0.03; all P < 0.05). A total of 1614 genes were changed in the SHR experiment, 1323 in the SHRSP, and 576 in the 2K1C. Eighty-eight genes were similarly regulated in all three models. Gene ontology enrichment analysis identified 59 ontologies that were enriched in all three datasets. These included over-representation to extracellular matrix, response to oxidative stress, and immune system processes. Out of the 88 in-common genes, 40 could be connected in a common pathway that was compared to two gene-expression profiles from human kidneys with histologically verified fibrosis to identify a highly significant number of in-common genes that were also represented in the common genetic pathway. There is a common pathway during the development of hypertensive kidney damage in rats irrespective of model. Interestingly, large parts of this common pathway are conserved in human kidney damage, which may indicate a broader importance in the development of chronic kidney disease.


Structural insights into complete metal ion coordination from ternary complexes of B family RB69 DNA polymerase.
We have captured a preinsertion ternary complex of RB69 DNA polymerase (RB69pol) containing the 3' hydroxyl group at the terminus of an extendable primer (ptO3') and a nonhydrolyzable 2'-deoxyuridine 5'-α,β-substituted triphosphate, dUpXpp, where X is either NH or CH(2), opposite a complementary templating dA nucleotide residue. Here we report four structures of these complexes formed by three different RB69pol variants with catalytically inert Ca(2+) and four other structures with catalytically competent Mn(2+) or Mg(2+). These structures provide new insights into why the complete divalent metal-ion coordination complexes at the A and B sites are required for nucleotidyl transfer. They show that the metal ion in the A site brings ptO3' close to the α-phosphorus atom (Pα) of the incoming dNTP to enable phosphodiester bond formation through simultaneous coordination of both ptO3' and the nonbridging Sp oxygen of the dNTP's α-phosphate. The coordination bond length of metal ion A as well as its ionic radius determines how close ptO3' can approach Pα. These variables are expected to affect the rate of bond formation. The metal ion in the B site brings the pyrophosphate product close enough to Pα to enable pyrophosphorolysis and assist in the departure of the pyrophosphate. In these dUpXpp-containing complexes, ptO3' occupies the vertex of a distorted metal ion A coordination octahedron. When ptO3' is placed at the vertex of an undistorted, idealized metal ion A octahedron, it is within bond formation distance to Pα. This geometric relationship appears to be conserved among DNA polymerases of known structure.


Inhibition of androgen metabolism and binding by a liposterolic extract of "Serenoa repens B" in human foreskin fibroblasts.
We previously suggested [Steroids 33, (1979) 3; Steroids 37, (1981) 6] that cultured genital skin fibroblasts should prove useful for screening of potential antiandrogens in human and living target cells. "Serenoa repens" lipidic extract (S.R.E.) was recently reported (Br. J. Pharmacol., in press) to inhibit androgen action in animals. The present investigation was designed to study the antiandrogenicity of this compound in human cells: we therefore analyzed the effects of S.R.E. on the intracellular conversion of testosterone (T) to 5 alpha-reduced derivatives, and we investigated interaction of S.R.E. with the intracellular androgen-receptor complex. Since the chemical structure of the active component of S.R.E. is still unknown, results are expressed in U/ml (one unit is defined as the amount of S.R.E. required to inhibit 50% of the specific binding (IC50) of [3H]1881 to rat prostate cytosol). S.R.E. at different dilutions (5.7 to 28.6 U/ml) is added to culture media containing [3H]T or [3H]dihydrotestosterone (DHT) and incubated at 37 degrees C with cultured fibroblasts. 28.6 U/ml S.R.E. significantly alters the formation of DHT and strongly inhibits 3 ketosteroid reductase mediated conversion of DHT to 5 alpha-androstane-3 alpha, 17 beta-diol, characterized radiochemically by thin-layer chromatography. S.R.E. is a good competitor for the whole cell androgen receptor: 7.1 U/ml S.R.E. gives 50% inhibition of the binding of 2 X 10(-9) M [3H]DHT to its receptor. Competitive binding assays after cell fractionation indicate that S.R.E. is less potent in nuclear than in cytosol receptors. Sucrose gradient centrifugation of the radioactive cell lysate of fibroblasts demonstrates that 28.6 U/ml S.R.E. abolishes 70% of the 3.6 S receptor-complex radioactive peak. The present studies show that S.R.E. inhibits 5 alpha-reductase, 3-ketosteroid reductase and receptor binding of androgens in cultured human foreskin fibroblasts. As the search for the ideal antiandrogen continues, S.R.E. appears to be a new type of antiandrogenic compound as therapeutics for the treatment of benign prostatic hypertrophy, hirsutism and so forth.


Insights into the phylogeny and coding potential of microbial dark matter.
Genome sequencing enhances our understanding of the biological world by providing blueprints for the evolutionary and functional diversity that shapes the biosphere. However, microbial genomes that are currently available are of limited phylogenetic breadth, owing to our historical inability to cultivate most microorganisms in the laboratory. We apply single-cell genomics to target and sequence 201 uncultivated archaeal and bacterial cells from nine diverse habitats belonging to 29 major mostly uncharted branches of the tree of life, so-called 'microbial dark matter'. With this additional genomic information, we are able to resolve many intra- and inter-phylum-level relationships and to propose two new superphyla. We uncover unexpected metabolic features that extend our understanding of biology and challenge established boundaries between the three domains of life. These include a novel amino acid use for the opal stop codon, an archaeal-type purine synthesis in Bacteria and complete sigma factors in Archaea similar to those in Bacteria. The single-cell genomes also served to phylogenetically anchor up to 20% of metagenomic reads in some habitats, facilitating organism-level interpretation of ecosystem function. This study greatly expands the genomic representation of the tree of life and provides a systematic step towards a better understanding of biological evolution on our planet.


Effects of two barrier creams on the diffusion of benzo[a]pyrene across human skin.
Barrier hand creams, often containing antiseptic agents, may provide a form of protection not only for health care professionals, but also for workers in the chemical and pharmaceutical industries. To evaluate the efficacy of two such barrier creams available on the South African market, the in vitro diffusion of a model compound, benzo[a]pyrene, through human skin at 20 degrees and 37 degrees C was studied. Treated (10 min) and untreated human skin disks (4 mm in diameter) were mounted in flow cells of a continuous flow-through diffusion apparatus. Buffer/tritiated benzo[a]pyrene was collected from the acceptor chambers at 2-hour intervals for a total of 24 hours and counted in a liquid scintillation counter. At 20 degrees C no significant differences could be detected between the flux rates of benzo[a]pyrene across barrier cream treated and untreated skin. However, at 37 degrees C Skinguard significantly increased flux rates of this carcinogen. Skin barrier creams therefore need to be carefully scrutinised with respect to their protective effects because the latter may vary for molecules with different chemical properties.


A polymetamorphic protein.
Arc repressor is a homodimeric protein with a ribbon-helix-helix fold. A single polar-to-hydrophobic substitution (N11L) at a solvent-exposed position leads to population of an alternate dimeric fold in which 3₁₀ helices replace a β-sheet. Here we find that the variant Q9V/N11L/R13V (S-VLV), with two additional polar-to-hydrophobic surface mutations in the same β-sheet, forms a highly stable, reversibly folded octamer with approximately half the α-helical content of wild-type Arc. At low protein concentration and low ionic strength, S-VLV also populates both dimeric topologies previously observed for N11L, as judged by NMR chemical shift comparisons. Thus, accumulation of simple hydrophobic mutations in Arc progressively reduces fold specificity, leading first to a sequence with two folds and then to a manifold bridge sequence with at least three different topologies. Residues 9-14 of S-VLV form a highly hydrophobic stretch that is predicted to be amyloidogenic, but we do not observe aggregates of higher order than octamer. Increases in sequence hydrophobicity can promote amyloid aggregation but also exert broader and more complex effects on fold specificity. Altered native folds, changes in fold coupled to oligomerization, toxic pre-amyloid oligomers, and amyloid fibrils may represent a near continuum of accessible alternatives in protein structure space.


Erosion of a B-Lynch suture through the uterine wall: a case report.
The B-Lynch uterine suture brace has been used for the surgical treatment of postpartum hemorrhage. To date, no complications of this procedure have been reported. We describe B-Lynch suture erosion through the uterine wall identified at a 6-week postpartum visit. A 19-year-old primigravida underwent a primary low transverse cesarean section at term for arrest of descent. The surgery was complicated by postpartum hemorrhage secondary to uterine atony unresponsive to medical management. The patient underwent successful placement of a B-Lynch suture using delayed, absorbable suture for control of the hemorrhage and had an uneventful postoperative course. At her 6-week postpartum examination, she was found to have the suture protruding from the uterine cervical os. The suture was removed in its entirety without difficulty. Follow-up sonohysterography at 6 months identified a small defect in the anterior wall of the lower uterine segment, corresponding to the probable site of suture erosion. Erosion of suture through the uterine wall can be a complication of the B-Lynch uterine suture brace. Delayed, absorbable suture is less desirable than absorbable suture for this procedure due to the risk of erosion through the uterine wall.


How intermittent breaks in interaction improve collective intelligence.
People influence each other when they interact to solve problems. Such social influence introduces both benefits (higher average solution quality due to exploitation of existing answers through social learning) and costs (lower maximum solution quality due to a reduction in individual exploration for novel answers) relative to independent problem solving. In contrast to prior work, which has focused on how the presence and network structure of social influence affect performance, here we investigate the effects of time. We show that when social influence is intermittent it provides the benefits of constant social influence without the costs. Human subjects solved the canonical traveling salesperson problem in groups of three, randomized into treatments with constant social influence, intermittent social influence, or no social influence. Groups in the intermittent social-influence treatment found the optimum solution frequently (like groups without influence) but had a high mean performance (like groups with constant influence); they learned from each other, while maintaining a high level of exploration. Solutions improved most on rounds with social influence after a period of separation. We also show that storing subjects' best solutions so that they could be reloaded and possibly modified in subsequent rounds-a ubiquitous feature of personal productivity software-is similar to constant social influence: It increases mean performance but decreases exploration.


Single-atom vibrational spectroscopy in the scanning transmission electron microscope.
Single-atom impurities and other atomic-scale defects can notably alter the local vibrational responses of solids and, ultimately, their macroscopic properties. Using high-resolution electron energy-loss spectroscopy in the electron microscope, we show that a single substitutional silicon impurity in graphene induces a characteristic, localized modification of the vibrational response. Extensive ab initio calculations reveal that the measured spectroscopic signature arises from defect-induced pseudo-localized phonon modes-that is, resonant states resulting from the hybridization of the defect modes and the bulk continuum-with energies that can be directly matched to the experiments. This finding realizes the promise of vibrational spectroscopy in the electron microscope with single-atom sensitivity and has broad implications across the fields of physics, chemistry, and materials science.


Quantification and Classification of E. coli Proteome Utilization and Unused Protein Costs across Environments.
The costs and benefits of protein expression are balanced through evolution. Expression of un-utilized protein (that have no benefits in the current environment) incurs a quantifiable fitness costs on cellular growth rates; however, the magnitude and variability of un-utilized protein expression in natural settings is unknown, largely due to the challenge in determining environment-specific proteome utilization. We address this challenge using absolute and global proteomics data combined with a recently developed genome-scale model of Escherichia coli that computes the environment-specific cost and utility of the proteome on a per gene basis. We show that nearly half of the proteome mass is unused in certain environments and accounting for the cost of this unused protein expression explains >95% of the variance in growth rates of Escherichia coli across 16 distinct environments. Furthermore, reduction in unused protein expression is shown to be a common mechanism to increase cellular growth rates in adaptive evolution experiments. Classification of the unused protein reveals that the unused protein encodes several nutrient- and stress- preparedness functions, which may convey fitness benefits in varying environments. Thus, unused protein expression is the source of large and pervasive fitness costs that may provide the benefit of hedging against environmental change.


The comparison between two irrigation regimens on the dentine wettability for an epoxy resin based sealer by measuring its contact angle formed to the irrigated dentine.
The aim was to assess the influence of two irrigation regimens having ethylenediaminetetraacetic acid (EDTA) and ethylenediaminetetraacetic acid with cetrimide (EDTAC) as final irrigants, respectively, on the dentine wettability for AH Plus sealer by comparing its contact angle formed to the irrigated dentine. Study samples were divided into two groups (n = 10). The groups were irrigated with 3% sodium hypochlorite (NaOCl) solution followed by either 17% EDTA or 17% EDTAC solution. AH Plus was mixed, and controlled volume droplet (0.1 mL) of the sealer was placed on the dried samples. The contact angle was measured using a Dynamic Contact Angle Analyzer and results were analyzed using SPSS 21.0 and 2 sample t-test. There was a significant difference in the contact angle of AH Plus formed to the dentine irrigated with the above two regimens. AH Plus showed significantly lower contact angle with the regimen having EDTAC as a final irrigant than the one with EDTA (P < 0.05). An irrigation regimen consisting of NaOCl with either EDTA or EDTAC solution as a final irrigant influences the dentine wettability and contact angle of a sealer. EDTAC as a final irrigant facilitates better dentin wettability than EDTA for AH Plus to promote its better flow and adhesion.


Distinct mechanisms of impairment in cognitive ageing and Alzheimer's disease.
Similar manifestations of functional decline in ageing and Alzheimer's disease obscure differences in the underlying cognitive mechanisms of impairment. We sought to examine the contributions of top-down attentional and bottom-up perceptual factors to visual self-movement processing in ageing and Alzheimer's disease. We administered a novel heading discrimination task requiring subjects to determine direction of simulated self-movement from left or right offset optic flow fields of several sizes (25 degrees, 40 degrees or 60 degrees in diameter) to 18 Alzheimer's disease subjects (mean age = 75.3, 55% female), 21 older adult control subjects (mean age = 72.4, 67% female), and 26 younger control subjects (mean age = 26.5, 63% female). We also administered computerized measures of processing speed and divided and selective attention, and psychophysical measures of visual motion perception to all subjects. Both older groups showed significant difficulty in judging the direction of virtual self-movement [F(2,194) = 40.5, P < 0.001] and optic flow stimulus size had little effect on heading discrimination for any group. Both older groups showed impairments on measures of divided [F(2,62) = 22.2, P < 0.01] and selective [F(2,62) = 63.0, P < 0.001] attention relative to the younger adult control group, while the Alzheimer's disease group showed a selective impairment in outward optic flow perception [F(2,64) = 6.3, P = 0.003] relative to both control groups. Multiple linear regression revealed distinct attentional and perceptual contributions to heading discrimination performance for the two older groups. In older adult control subjects, poorer heading discrimination was attributable to attentional deficits (R(2) adj = 0.41, P = 0.001) whereas, in Alzheimer's disease patients, it was largely attributable to deficits of visual motion perception (R(2) adj = 0.57, P < 0.001). These findings suggest that successive attentional and perceptual deficits play independent roles in the progressive functional impairments of ageing and Alzheimer's disease. We speculate that the attentional deficits that dominate in older adults may promote the development of the perceptual deficits that further constrain performance in Alzheimer's disease.


Neuropeptides in tendinopathy.
Overuse tendinopathy remains a major clinical burden for sports medicine and general practitioners. Recent studies have highlighted the role of sensory and autonomic nerves in generating or perpetuating the symptoms and tissue abnormalities associated with tendinopathy. We outline the neuroanatomy and potential roles of nerves and associated neuropeptides in tendinopathy. In addition, intriguing new data is reviewed which suggests that there may be a substantial intrinsic source of neuropeptides within tendons - namely, the tenocytes themselves. The potential roles of Substance P and mast cells are highlighted in particular. We discuss the implications for conservative management including sclerosing injections and exercise training.


Corticosteroid injection for shoulder pain: single-blind randomized pilot trial in primary care.
Shoulder pain is a very common presentation in primary care. Evidence of benefit for subacromial corticosteroid injection is inconclusive and confined largely to studies with short follow-up. We plan a large, definitive, primary-care-based trial to determine efficacy and safety in patients with rotator cuff tendinopathy, and conducted a pilot trial to explore feasibility. Six general practitioners (GPs) from Oxfordshire, UK underwent update training in assessing painful shoulders and injecting the subacromial space. Each then recruited patients aged 35 to 74 years from primary care complaining of shoulder pain lasting no more than 6 months. Eligible participants were randomized to receive either methylprednisolone acetate 40 mg with lidocaine 1% (total volume 1 ml), or lidocaine 1% alone (total volume 1 ml), injected into the subacromial space. The participants were blinded to treatment allocation. Feasibility outcomes were rates of recruitment, withdrawal, adherence to the protocol, completeness of follow-up, and success of patient masking. Clinical outcomes were the Oxford Shoulder Score (OSS) at baseline and at 4 and 12 weeks, and responses to three satisfaction questions at 2, 4 and 12 weeks. Outcome data were collected by postal questionnaires. A total of 40 participants were randomized (80% of the target 50 participants) over 26 weeks giving an overall recruitment rate of 1.5 participants per week. Rates of follow-up were maintained to a high level for the full 12 weeks. Four participants requested a 'rescue' corticosteroid injection but no patients withdrew. The trial GPs gave high scores for their confidence that the patient had remained blinded to treatment allocation during the procedure. The OSS at 4 and 12 weeks and the responses to the satisfaction questions are reported. It is feasible to recruit participants with shoulder pain in the primary care setting for a blinded, randomized trial of corticosteroid injection. Online randomization of participants from the practice is also feasible, and postal questionnaires provide an effective means of gathering outcome data in this area of study. The lessons learned from this pilot will usefully inform the design of a large, definitive efficacy trial in primary care. Current Clinical Trials ISRCTN82357435.


Diagnostic potential of Western blot analysis of sera from dogs with leishmaniasis in endemic areas and significance of the pattern.
Serum samples collected from 237 dogs in Catalonia (northeastern Spain) were screened by Western blot analysis to detect the presence of antibodies specific to different Leishmania infantum polypeptide fractions. Leishmaniasis was confirmed in 72 of these dogs by direct examination and/or culture. Another 165 animals from the Priorat region were studied periodically for 2-8 years between 1987 and 1995, giving a total of 565 determinations. A control group of 93 dogs from nonendemic areas was also studied. Sera from dogs with leishmaniasis recognized antigens with molecular weights ranging from 12 to 85 kD. The most sensitive antigens were those of 70, 65, 46, 30, 28, 14, and 12 kD, which were recognized by 75%, 75%, 78%, 75%, 81%, 79%, and 75%, respectively, of the sera from dogs with positive parasitologic examination results. Antigens of 70 and 65 kD were also recognized by two dogs from nonendemic areas. Antigens of 14 and 12 kD were the first to be recognized by sera of asymptomatic dogs with titers less than the cut-off value of the dot-ELISA that increased during the longitudinal study, and the presence of antibodies specific for these fractions was observed for up to six years before seroconversion observed by dot-ELISA. These antibodies were also the first to disappear in dogs in which the disease was self-limited. The study corroborates the high sensitivity and specificity of Western blots in the diagnosis of canine leishmaniasis when the bands of low molecular weight (less than 46 kD) are considered, and indicates that fractions of 14 and 12 kD are useful in detecting early forms of the disease.


Behavioral and neural valuation of foods is driven by implicit knowledge of caloric content.
The factors that affect food choices are critical to understanding obesity. In the present study, healthy participants were shown pictures of foods to determine the impact of caloric content on food choice. Brain activity was then measured while participants bid for a chance to purchase and eat one item. True caloric density, but not individual estimates of calorie content, predicted how much participants were willing to pay for each item. Caloric density also correlated with the neural response to food pictures in the ventromedial prefrontal cortex, a brain area that encodes the value of stimuli and predicts immediate consumption. That same region exhibited functional connectivity with an appetitive brain network, and this connectivity was modulated by willingness to pay. Despite the fact that participants were poor at explicitly judging caloric content, their willingness to pay and brain activity both correlated with actual caloric density. This suggests that the reward value of a familiar food is dependent on implicit knowledge of its caloric content.


Osteoporosis in treated adult coeliac disease.
Forty five women and 10 men with coeliac disease diagnosed in adult life, who were already on a gluten free diet, had serial bone mineral density measurements at the lumbar spine and femoral neck over 12 months. Osteoporosis, defined as a bone mineral density (BMD) < or = 2 SD below the normal peak bone mass was found in 50% of male and 47% of female coeliac patients. Patients with a BMD < or = 2 SD below age and sex matched normal subjects, had a significantly lower body mass index (21.3 kg.m-2 compared with 25.2 kg.m-2, p < 0.02 Wilcoxon rank sum test) and lower average daily calcium intake (860 mg/day compared with 1054 mg/day, p < 0.05 Wilcoxon rank sum test) than patients with normal bone mineral density. In postmenopausal women with coeliac disease there was a strong correlation between the age at menopause and BMD at both the lumbar spine (r = 0.681, p < 0.01, Spearman's rank correlation) and femoral neck (r = 0.632, p < 0.01). No overall loss of bone was shown over the 12 months of follow up, and relative to the reference population there was a significant improvement in BMD at the lumbar spine in women (p < 0.025, paired t test) and at the femoral neck in men (p < 0.05, paired t test). There was a significant negative correlation between the annual percentage change in BMD at the lumbar spine and the duration of gluten free diet (r = -0.429, p<0.01, Spearman's rank correlation), with the largest gain in BMD in patients with most recently diagnosed coeliac disease. Osteoporosis was shown in 47% of patients with treated adult coeliac disease. Recognised risk factors for osteoporosis in the general population including low body mass index, dietary calcium intake, and early menopause are particularly important in coeliac disease. Treatment of coeliac disease with a gluten free diet probably protects against further bone loss, and in the early stages is associated with a gain in bone mineral density.


Refined flicker photometry technique to measure ocular lens density.
Many physiological and pathological conditions are associated with a change in the crystalline lens transmittance. Estimates of lens opacification, however, generally rely on subjective rather than objective measures in clinical practice. The goal of our study was to develop an improved psychophysical heterochromatic flicker photometry technique combined with existing mathematical models to evaluate the spectral transmittance of the human ocular media noninvasively. Our results show that it is possible to accurately estimate ocular media density in vivo in humans. Potential applications of our approach include basic research and clinical settings on visual and nonimage-forming visual systems.


Review of the fish-parasitic genus Ceratothoa Dana, 1852 (Crustacea: Isopoda: Cymothoidae) from Australia, with description of two new species.
The genus Ceratothoa Dana, 1852, is revised for Australian waters. Ceratothoa is represented in Australia by nine species, including two new species: Ceratothoa barracuda sp. nov. described from Cairns and Ceratothoa globulus sp. nov. described from Lord Howe Island. Ceratothoa imbricata Fabricius, 1775 is redescribed, with Ceratothoa trillesi (Avdeev, 1979) and Ceratothoa huttoni Filhol, 1885 placed into junior synonymy; the preferred hosts are species of the genus Trachurus (Carangidae). Ceratothoa banksii (Leach, 1818) is validated and brought out of synonymy with Ceratothoa imbricata; host species are from the families Kyphosidae, Scombridae, Latridae, Carangidae, Mugilidae, Salmonidae, Scatophagidae, Pomatomidae and Hemiramphidae. Species excluded from the Australian fauna are Ceratothoa trigonocephala (Leach, 1818) with an unknown host identity and type locality; and Ceratothoa lineata Miers, 1876a, that here is transferred to the genus Mothocya Costa, 1851, with Mothocya ihi Bruce, 1986 placed into junior synonymy. Ceratothoa contracta (Miers, 1880), the New Zealand Ceratothoa novaezelandiae Filhol, 1885 and the East Pacific Ceratothoa gaudichaudii (Milne Edwards, 1840) are regarded here as species inquirenda. A key to the Australian species of Ceratothoa is presented.


Antigens in tea-beverage prime human Vgamma 2Vdelta 2 T cells in vitro and in vivo for memory and nonmemory antibacterial cytokine responses.
Human gammadelta T cells mediate innate immunity to microbes via T cell receptor-dependent recognition of unprocessed antigens with conserved molecular patterns. These nonpeptide alkylamine antigens are shared by tumor cells, bacteria, parasites, and fungi but also by edible plant products such as tea, apples, mushrooms, and wine. Here we show that priming of gammadelta T cells with alkylamine antigens in vitro results in a memory response to these antigens. Such priming results also in a nonmemory response to whole bacteria and to lipopolysaccharide, characterized by IL-12-dependent secretion of IFN-gamma by gammadelta T cells and by gammadelta T cell proliferation. Drinking tea, which contains l-theanine, a precursor of the nonpeptide antigen ethylamine, primed peripheral blood gammadelta T cells to mediate a memory response on reexposure to ethylamine and to secrete IFN-gamma in response to bacteria. This unique combination of innate immune response and immunologic memory shows that gammadelta T cells can function as a bridge between innate and acquired immunity. In addition, these data provide an explanation for the health benefits of tea.