Intuition-Enabled Machine Learning Beats the Competition When Joint Human-Robot Teams Perform Inorganic Chemical Experiments.
Traditionally, chemists have relied on years of training and accumulated experience in order to discover new molecules. But the space of possible molecules is so vast that only a limited exploration with the traditional methods can be ever possible. This means that many opportunities for the discovery of interesting phenomena have been missed, and in addition, the inherent variability of these phenomena can make them difficult to control and understand. The current state-of-the-art is moving toward the development of automated and eventually fully autonomous systems coupled with in-line analytics and decision-making algorithms. Yet even these, despite the substantial progress achieved recently, still cannot easily tackle large combinatorial spaces, as they are limited by the lack of high-quality data. Herein, we explore the utility of active learning methods for exploring the chemical space by comparing the collaboration between human experimenters with an algorithm-based search against their performance individually to probe the self-assembly and crystallization of the polyoxometalate cluster Na6[Mo120Ce6O366H12(H2O)78]·200H2O (1). We show that the robot-human teams are able to increase the prediction accuracy to 75.6 ± 1.8%, from 71.8 ± 0.3% with the algorithm alone and 66.3 ± 1.8% from only the human experimenters demonstrating that human-robot teams can beat robots or humans working alone.


An ultrasensitive assay format for detecting ULK1 inhibition by monitoring the phosphorylation status of Atg13.
A new technology from Quanterix called SiMoA (single molecule array) which employs a fully automated system capable of ultrasensitive sandwich based ELISA detection was explored. Our studies focused upon the inhibition of the autophagy initiating kinase ULK1 by measuring the both total Atg13 and the phosphorylation of Atg13(pSer(318)) from control and following compound treatment in either overexpressing or wild type tissue culture samples. The results show linear protein concentration dependence over two orders of magnitude and provide an assay window of 8- to 100-fold signal to background for inhibition of phosphorylation for both wild type and overexpressed samples, respectively. Moreover, overexpressed samples displayed 17-fold pSer(318)-Atg13 above wild type levels of with no apparent differences in compound potency. Lastly, the inhibition of ULK1 from mouse derived wild type xenografts also demonstrated loss of pSer(318)-Atg13 upon ULK1 inhibitor treatment that compared favorably to Western blot. These results show that the SiMoA technology can detect quantitatively low levels of endogenous biomarkers with the ability to detect the loss of pSer(318)-Atg13 upon ULK1 inhibition.


Evolution of a programme to engage school students with health research and science in Kenya.
Facilitating mutually-beneficial educational activities between researchers and school students is an increasingly popular way for research institutes to engage with communities who host health research, but these activities have rarely been formally examined as a community or public engagement approach in health research. The KEMRI-Wellcome Trust Research Programme (KWTRP) in Kilifi, Kenya, through a Participatory Action Research (PAR) approach involving students, teachers, researchers and education stakeholders, has incorporated 'school engagement' as a key component into their community engagement (CE) strategy. School engagement activities at KWTRP aim at strengthening the ethical practice of the institution in two ways: through promoting an interest in science and research among school students as a form of benefit-sharing; and through creating forums for dialogue aimed at promoting mutual understanding between researchers and school students. In this article, we provide a background of CE in Kilifi and describe the diverse ways in which health researchers have engaged with communities and schools in different parts of the world. We then describe the way in which the KWTRP school engagement programme (SEP) was developed and scaled-up. We conclude with a discussion about the challenges, benefits and lessons learnt from the SEP implementation and scale-up in Kilifi, which can inform the establishment of SEPs in other settings.


Fully automated, semiautomated, and manual morphometric analysis of corneal subbasal nerve plexus in individuals with and without diabetes.
The aim of the study was to determine the association, agreement, and detection capability of manual, semiautomated, and fully automated methods of corneal nerve fiber length (CNFL) quantification of the human corneal subbasal nerve plexus (SNP). Thirty-three participants with diabetes and 17 healthy controls underwent laser scanning corneal confocal microscopy. Eight central images of the SNP were selected for each participant and analyzed using manual (CCMetrics), semiautomated (NeuronJ), and fully automated (ACCMetrics) software to quantify the CNFL. For the entire cohort, mean CNFL values quantified by CCMetrics, NeuronJ, and ACCMetrics were 17.4 ± 4.3 mm/mm, 16.0 ± 3.9 mm/mm, and 16.5 ± 3.6 mm/mm, respectively (P < 0.01). CNFL quantified using CCMetrics was significantly higher than those obtained by NeuronJ and ACCMetrics (P < 0.05). The 3 methods were highly correlated (correlation coefficients 0.87-0.98, P < 0.01). The intraclass correlation coefficients were 0.87 for ACCMetrics versus NeuronJ and 0.86 for ACCMetrics versus CCMetrics. Bland-Altman plots showed good agreement between the manual, semiautomated, and fully automated analyses of CNFL. A small underestimation of CNFL was observed using ACCMetrics with increasing the amount of nerve tissue. All 3 methods were able to detect CNFL depletion in diabetic participants (P < 0.05) and in those with peripheral neuropathy as defined by the Toronto criteria, compared with healthy controls (P < 0.05). Automated quantification of CNFL provides comparable neuropathy detection ability to manual and semiautomated methods. Because of its speed, objectivity, and consistency, fully automated analysis of CNFL might be advantageous in studies of diabetic neuropathy.


Beneficial Effects of a Dietary Weight Loss Intervention on Human Gut Microbiome Diversity and Metabolism Are Not Sustained during Weight Maintenance.
In the present study, we examined the effect of a very low-calorie diet(VLCD)-based obesity program on human gut microbiome diversity and metabolism during weight loss and weight maintenance. Obese subjects underwent 3 months of VLCD followed by 3 months of weight maintenance. A lean and an obese control group were included. The microbiome was characterized by performing high-throughput dual-indexed 16S rDNA amplicon sequencing. At baseline, a significant difference in the Firmicutes/Bacteroidetes ratio between the lean and obese individuals was observed (p = 0.047). The VLCD resulted in significant alterations in gut microbiome diversity from baseline to 3 months (p = 0.0053). Acinetobacter represented an indicator species for the observed effect (indicator value = 0.998, p = 0.006). Metabolic analyses revealed alterations of the bacterial riboflavin pathway from baseline to 3 months (pnom = 0.0078). These changes in diversity and bacterial metabolism induced by VLCD diminished during the weight maintenance phase, despite sustained reductions in body weight and sustained improvements of insulin sensitivity. The present data show that a VLCD is able to beneficially alter both gut microbiome diversity and metabolism in obese humans, but that these changes are not sustained during weight maintenance. This finding might suggest that the microbiome should be targeted during obesity programs.


Electronic transport in nanoparticle monolayers sandwiched between graphene electrodes.
Graphene/CdSe nanoparticle monolayer/graphene sandwich structures were fabricated to explore the interactions between these layered materials. Electrical transport across these heterostructures suggests that transport is limited by tunneling through the nanoparticle (NP) ligands but not the NP core itself. Photoconductivity suggests ligands may affect the exciton separation efficiency.


The Effect of Omega-3 and Omega-6 Polyunsaturated Fatty Acids on the Production of Cyclooxygenase and Lipoxygenase Metabolites by Human Umbilical Vein Endothelial Cells.
Although the correlation between polyunsaturated fatty acids (PUFA) and the production of pro- and anti-inflammatory metabolites is well documented, little is known about the simultaneous effect of different PUFA on the production of cyclooxygenase and lipoxygenase metabolites. The present research examines the association between different omega-3 (ω-3) and omega-6 (ω-6) PUFA and the release of four cyclooxygenase and six lipoxygenase metabolites in cell medium by human umbilical vein endothelial cells (HUVEC). The different combinations of ω-3 and ω-6 PUFA were prepared according to a full 24 factorial design that enables studying not only the main effects but also the different interactions between fatty acids. In addition, interactions diagrams and principal component analysis were useful tools for interpreting higher order interactions. To the best of our knowledge, this is the first report addressing the combined effect of ω-3 and ω-6 PUFA on the signaling of prostaglandins, prostacyclins, leukotrienes and resolvins by HUVEC.


Stability of nanofluids in quiescent and shear flow fields.
An experimental study was conducted to investigate the structural stability of ethylene glycol-based titanium dioxide nanoparticle suspensions (nanofluids) prepared by two-step method. The effects of particle concentration, fluid temperature, shear rate and shear duration were examined. Particle size and thermal conductivity measurements in quiescent state indicated the existence of aggregates and that they were stable in temperatures up to 60°C. Shear stability tests suggested that the structure of nanoparticle aggregates was stable in a shear interval of 500-3000 s-1 measured over a temperature range of 20-60°C. These findings show directions to resolve controversies surrounding the underlying mechanisms of thermal conduction and convective heat transfer of nanofluids.


Clinicopathological study of livers from brain-dead patients treated with a combination of vasopressin and epinephrine.
Studies were made on the pathological lesions and biochemical indices of the livers of 22 patients in whom normal hemodynamics was maintained for 0-48 days after brain death by administration of vasopressin and epinephrine. Thirty-one specimens of liver tissues were obtained by percutaneous biopsy or at autopsy. The degrees of central venous congestion, central fibrosis, focal fibrosis, fatty metamorphosis, piecemeal necrosis, periportal fibrosis, and intrahepatic cholangitis in livers on various days after brain death were compared with those on the day of brain death (day 0). Central venous congestion was extensive on days 0-4, significantly less on days 5-14, and then again extensive on days 15-48. Central fibrosis and focal fibrosis showed no remarkable change during the 48-day period. Fatty metamorphosis, piecemeal necrosis, and periportal fibrosis showed no significant changes until day 16, but spread extensively on days 40-48. Intrahepatic cholangitis was scarcely observed on day 0 but began to increase after day 3, and spread extensively after day 5. The level of serum glutamic pyruvic transaminase did not increase in most patients until day 15. The mean value of prothrombin activity also did not decrease until day 15. However, the mean value of serum alkaline phosphatase increased gradually after day 3, and was correlated with cholangitis. The present study showed that during prolonged hemodynamic maintenance of brain-dead patients, pathological lesions did not spread or diminished and that biochemical indices did not become worse, or improved, in the first 2 weeks, except for increases in cholangitis and the serum alkaline phosphatase level.


Association of the T-cell protein tyrosine phosphatase with nuclear import factor p97.
Alternative splicing of the T-cell protein tyrosine phosphatase (TCPTP) transcript generates two forms of the enzyme that differ at their extreme C termini: a 48-kDa endoplasmic reticulum-associated form and a 45-kDa nuclear form. By affinity chromatography, using GST-TCPTP fusion proteins, we have isolated three cytoplasmic proteins of 120, 116, and 97 kDa that interact with TCPTP. The p120 protein associated with residues 377-415 from the C terminus of the 48-kDa form of TCPTP, whereas the recognition site for p97 and p116 was mapped to residues 350-381 encompassing the TCPTP nuclear localization sequence (NLS). The TCPTP NLS was shown to be bipartite, requiring basic residues 350-358 (basic cluster I) and 377-381 (basic cluster II), the sites of interaction with p97 and p116, for efficient nuclear translocation. The interaction between p97, p116, and the TCPTP NLS appeared unique in that these proteins did not form a stable interaction with the classical NLS of SV40 large T antigen or the standard bipartite NLS of nucleoplasmin. Sequence analysis of p97 identified it as the nuclear import factor p97 (importin-beta), which is an essential component of the nuclear import machinery. In assays in vitro in permeabilized cells, p97 was necessary but not sufficient for optimal nuclear import of TCPTP. We found that TCPTP co-immunoprecipitated with the nuclear import factor p97 from cell lysates and that purified recombinant p97 and TCPTP interacted directly in vitro. These results indicate selectivity in the binding of p97 and p116 to the TCPTP NLS and suggest that p97 may mediate events that are distinct from the classical nuclear import process. Moreover, these results demonstrate that the C-terminal segment of TCPTP contains docking sites for interaction with proteins that may function to target the enzyme to defined intracellular locations and in the process regulate TCPTP function.


Implementation of the Abortion Act: report on a year's working of abortion clinics and operating sessions.
The results of the first year's working of abortion clinics and operating sessions are reported. Seventy-eight per cent. of patients were managed on the basis of day admission and 20% were able to leave the hospital within four hours, operation having been carried out by either Karman catheter or by Bierer suction curette under paracervical block. These results were possible because of quick reference of patients to the clinics and the absence of delay in admission for operation.The measures detailed go some way towards managing the abortion problem within the hospital service; similar arrangements could be instituted elsewhere by the responsible regional hospital boards.


Estimating energy expenditure using body-worn accelerometers: a comparison of methods, sensors number and positioning.
Several methods to estimate energy expenditure (EE) using body-worn sensors exist; however, quantifications of the differences in estimation error are missing. In this paper, we compare three prevalent EE estimation methods and five body locations to provide a basis for selecting among methods, sensors number, and positioning. We considered 1) counts-based estimation methods, 2) activity-specific estimation methods using METs lookup, and 3) activity-specific estimation methods using accelerometer features. The latter two estimation methods utilize subsequent activity classification and EE estimation steps. Furthermore, we analyzed accelerometer sensors number and on-body positioning to derive optimal EE estimation results during various daily activities. To evaluate our approach, we implemented a study with 15 participants that wore five accelerometer sensors while performing a wide range of sedentary, household, lifestyle, and gym activities at different intensities. Indirect calorimetry was used in parallel to obtain EE reference data. Results show that activity-specific estimation methods using accelerometer features can outperform counts-based methods by 88% and activity-specific methods using METs lookup for active clusters by 23%. No differences were found between activity-specific methods using METs lookup and using accelerometer features for sedentary clusters. For activity-specific estimation methods using accelerometer features, differences in EE estimation error between the best combinations of each number of sensors (1 to 5), analyzed with repeated measures ANOVA, were not significant. Thus, we conclude that choosing the best performing single sensor does not reduce EE estimation accuracy compared to a five sensors system and can reliably be used. However, EE estimation errors can increase up to 80% if a nonoptimal sensor location is chosen.


Malaria on the World Wide Web.
The Internet is enabling scientists and clinicians in areas with endemic malaria to transfer information to scientists and clinicians in other countries. This should allow changes in therapy to follow the rapid changes in the disease that have posed such difficulties in the past. This article reviews Internet resources that focus on malaria. This includes 90 Web sites in 12 sections. Authoritative multinational organizational sites and regional sites, such as those in Africa, Asia (including Thailand and India), and South America (in Venezuela and Brazil), are described. Basic research-oriented databases, such as those that deal with plasmodia genomics, biochemistry, and vaccine development, as well as vector information and geographic satellite information systems, are reviewed. There is a section about malaria research-funding organizations that offer online applications. Useful teaching resources and journals, including those with full online access, are detailed.


A 980 nm laser-activated upconverted persistent probe for NIR-to-NIR rechargeable in vivo bioimaging.
Long-lasting persistent luminescent nanoparticles (PLNPs) with efficient near-infrared (NIR) emission have emerged as a new generation of probes for in vivo optical bioimaging owing to their advantages of zero-autofluorescence benefited from the self-sustained emission after excitation, deep penetration depth, and a high signal-to-noise ratio. However, most of the PLNPs are charged by ultraviolet (UV) or visible light, remarkably limiting their applications for in vivo long-term bioimaging. Here we demonstrate 980 nm laser activated upconversion-PLNPs (UC-PLNPs) with efficient NIR emission. The NIR-emitting UC-PLNPs (Zn3Ga2GeO8:Yb/Er/Cr) were synthesized by a sol-gel method with subsequent calcination. Owing to the efficient energy-transfer between Er and Cr ions, these UC-PLNPs present long-lasting up to 15 h NIR emission at 700 nm after the excitation of a 980 nm laser; in which both excitation and emission bands fall within the biological transparent window. The results of in vitro/in vivo toxicity assessments indicate that UC-PLNPs after surface modification present low biotoxicity and side effects in living animals. More importantly, the synthesized UC-PLNPs can be effectively recharged by 980 nm laser to restore in vivo persistent bioimaging signals and can also be employed as nanoprobes for in vivo UC optical bioimaging. This is the first demonstration of rechargeable UC-PLNPs for NIR-to-NIR in vivo bioimaging. We believe that the synthesized UC-PLNPs by combining UC and persistent luminescence properties into a single host may have potential applications in the bioimaging area and pave the way for widely using PLNPs for in vivo renewable long-lasting bioimaging.


Epizootic activity of Murray Valley encephalitis and Kunjin viruses in an aboriginal community in the southeast Kimberley region of Western Australia: results of mosquito fauna and virus isolation studies.
We undertook annual surveys of flavivirus activity in the community of Billiluna in the southeast Kimberley region of Western Australia between 1989 and 2001 [corrected]. Culex annulirostris was the dominant mosquito species, particularly in years of above average rains and flooding. Murray Valley encephalitis (MVE) virus was isolated in 8 of the 13 years of the study from seven mosquito species, but more than 90% of the isolates were from Cx. annulirostris. The results suggest that MVE virus is epizootic in the region, w ith activity only apparent in years with average or above average rainfall and increased numbers of Cx. annulirostris. High levels of MVE virus activity and associated human cases were detected only once (in 1993) during the survey period. Activity of MVE virus could only be partially correlated with wet season rainfall and flooding, suggesting that a number of other factors must also be considered to accurately predict MVE virus activity at such communities.


Impact of malnutrition on propofol consumption and recovery time among patients undergoing laparoscopic gastrointestinal surgery.
Malnutrition is a major health problem, especially in hospitalized patients as it can be closely related to many post-operative complications. However, research on malnutrition and its effect on the outcome of general anesthesia have been largely neglected. Here we investigated malnutrition status on propofol consumption and recovery time among patients undergoing laparoscopic gastrointestinal surgery under general anesthesia. One hundred and one patients were recruited between January and June 2012 at Tongji Hospital and assigned into three groups according to Nutritional Risk Screening Tool 2002 score. A standard combined general anesthesia procedure was performed under regular monitoring. The dosage of propofol needed for induction, consumption during maintenance and recovery time were recorded. When compared with normal nutritional status individuals, the propofol dosage at induction was significantly decreased about 4.3% in moderate malnutritional status patients (P < 0.01) and about 16.8% in severely malnutritional status patients (P < 0.01). The average consumption of propofol was also significantly lower in malnourished individuals; for moderate malnutritional, the decrease was about 20% (P < 0.01) while for the severely malnutritional, it was 30% (P < 0.01) when compared with normal nutritional status individuals. For the recovery time of propofol anesthesia, the patients with severe malnutritional status awoke average 6.8 min later than those normally nourished (P < 0.01), but those patients with moderate malnutrition status did not (P = 0.885). The present results indicate that the dosage and recovery time of propofol does change in malnourished individuals. Therefore, malnutrition may somehow affect the outcome of general anesthesia.


Psychosocial dimensions of SLE: implications for the health care team.
The purpose of this exploratory study was threefold, ie, to clarify the unique psychosocial challenges facing those living with systemic lupus erythematosus (SLE), to distinguish which sociodemographic variables impact the lives of SLE patients, and generate knowledge regarding the way patients perceive SLE medication regimens. This was a cross-sectional exploratory study in 378 patients diagnosed with SLE and receiving services from the SLE Lupus Foundation in New York City. In addition to sociodemographic variables, the instrument used consisted of two scales, ie, the Systemic Lupus Erythematosus Needs Questionnaire (SLENQ) and the Multidimensional Health Locus of Control Scale, as well as questions regarding subjective perceptions of side effects from SLE medication. The highest general cause of self-reported depressive and anxious feelings was changes in appearance due to SLE, and limitations in physical abilities due to SLE (primarily from muscle and joint pain). The higher the sense of control over SLE, the less likely respondents were to report feeling depressed and anxious. African-American and Hispanic SLE patients reported a higher level of unmet psychological needs due to SLE than did their other ethnic counterparts. Weight gain and hair loss were the most likely medication side effects and also the most likely causes of SLE-related depression and anxiety. Those living with SLE are at risk for feelings of depression and anxiety. African-American and Hispanic women are at higher risk for these emotional states. Comprehensive assessment across the disciplines should screen this group of patients for depression and anxiety, and be prepared to refer them to patient education and social work counseling as indicated.


Transport of ER vesicles on actin filaments in neurons by myosin V.
Axoplasmic organelles in the giant axon of the squid have been shown to move on both actin filaments and microtubules and to switch between actin filaments and microtubules during fast axonal transport. The objectives of this investigation were to identify the specific classes of axoplasmic organelles that move on actin filaments and the myosin motors involved. We developed a procedure to isolate endoplasmic reticulum (ER) from extruded axoplasm and to reconstitute its movement in vitro. The isolated ER vesicles moved on exogenous actin filaments adsorbed to coverslips in an ATP-dependent manner without the addition of soluble factors. Therefore myosin was tightly bound and not extracted during isolation. These vesicles were identified as smooth ER by use of an antibody to an ER-resident protein, ERcalcistorin/protein disulfide isomerase (EcaSt/PDI). Furthermore, an antibody to squid myosin V was used in immunogold EM studies to show that myosin V localized to these vesicles. The antibody was generated to a squid brain myosin (p196) that was classified as myosin V based on comparisons of amino acid sequences of tryptic peptides of this myosin with those of other known members of the myosin V family. Dual labeling with the squid myosin V antibody and a kinesin heavy chain antibody showed that the two motors colocalized on the same vesicles. Finally, antibody inhibition experiments were performed with two myosin V-specific antibodies to show that myosin V motor activity is required for transport of vesicles on actin filaments in axoplasm. One antibody was made to a peptide in the globular tail domain and the other to the globular head fragment of myosin V. Both antibodies inhibited vesicle transport on actin filaments by greater than 90% compared to controls. These studies provide the first direct evidence that ER vesicles are transported on actin filaments by myosin V. These data confirm the role of actin filaments in fast axonal transport and provide support for the dual filament model of vesicle transport.


High-efficiency recombinant protein purification using mCherry and YFP nanobody affinity matrices.
Mammalian cell lines are important expression systems for large proteins and protein complexes, particularly when the acquisition of post-translational modifications in the protein's native environment is desired. However, low or variable transfection efficiencies are challenges that must be overcome to use such an expression system. Expression of recombinant proteins as a fluorescent protein fusion enables real-time monitoring of protein expression, and also provides an affinity handle for one-step protein purification using a suitable affinity reagent. Here, we describe a panel of anti-GFP and anti-mCherry nanobody affinity matrices and their efficacy for purification of GFP/YFP or mCherry fusion proteins. We define the molecular basis by which they bind their target proteins using X-ray crystallography. From these analyses, we define an optimal pair of nanobodies for purification of recombinant protein tagged with GFP/YFP or mCherry, and demonstrate these nanobody-sepharose supports are stable to many rounds of cleaning and extended incubation in denaturing conditions. Finally, we demonstrate the utility of the mCherry-tag system by using it to purify recombinant human topoisomerase 2α expressed in HEK293F cells. The mCherry-tag and GFP/YFP-tag expression systems can be utilized for recombinant protein expression individually or in tandem for mammalian protein expression systems where real-time monitoring of protein expression levels and a high-efficiency purification step is needed.


Menaquinone-7 regulates the expressions of osteocalcin, OPG, RANKL and RANK in osteoblastic MC3T3E1 cells.
Epidemiological studies show that dietary intake of natto, which contains significant amount of vitamin K(2), reduces the risk of bone formation loss. However, many confounding factors, such as calcium and isoflavone, are found in natto, because it is made from soybeans. In this study, the direct effects of MK-7, a vitamin K(2) analogue, were assessed in osteoblasts. Osteoblastic MC3T3E1 cells were cultured with or without MK-7 for 10 days and the number of cells was calculated. The cell count was not different between MK-7 treated cells and control cells for 1, 2, and 4 days. However, it was significantly suppressed in MK-7 treated cells at 10 days, suggesting that MK-7 suppressed cell proliferation. Real-time PCR analysis showed that mRNAs of osteocalcin (OC), osteoprotegerin (OPG), and the receptor activator of the NFkappaB ligand (RANKL) were induced after MK-7 administration to the culture medium. RANK mRNA expression was also enhanced by MK-7 administration. Immunocytochemical analysis showed that MK-7 increased the protein levels of OC and RANKL. RANK protein was also enhanced, but this induction was suppressed by anti-RANK antibody administration. This suppression was recovered when anti-RANK antibody and MK-7 were administered. These observations suggest that MK-7 may directly affect MC3T3E1 cells and stimulate osteoblastic differentiation, not proliferation.


Bis-Quinolinium Cyclophane Blockers of SK Potassium Channels Are Antagonists of M3 Muscarinic Acetylcholine Receptors.
Dequalinium is used as an antimicrobial compound for oral health and other microbial infections. Derivatives of dequalinium, the bis-quinolinium cyclophanes UCL 1684 and UCL 1848, are high affinity SK potassium channel antagonists. Here we investigated these compounds as M3 muscarinic receptor (mACHR) antagonists. We used the R-CEPIAer endoplasmic reticulum calcium reporter to functionally assay for Gq-coupled receptor signaling, and investigated the bis-quinolinium cyclophanes as antagonists of M3 mACHR activation in transfected CHO cells. Given mACHR roles in airway smooth muscle (ASM) contractility, we also tested the ability of UCL 1684 to relax ASM. We find that these compounds antagonized M3 mACHRs with an IC50 of 0.27 μM for dequalinium chloride, 1.5 μM for UCL 1684 and 1.0 μM for UCL 1848. UCL 1684 also antagonized M1 (IC50 0.12 μM) and M5 (IC50 0.52 μM) mACHR responses. UCL 1684 was determined to be a competitive antagonist at M3 receptors as it increased the EC50 for carbachol without a reduction in the maximum response. The Ki for UCL1684 determined from competition binding experiments was 909 nM. UCL 1684 reduced carbachol-evoked ASM contractions (>90%, IC50 0.43 μM), and calcium mobilization in rodent and human lung ASM cells. We conclude that dequalinium and bis-quinolinium cyclophanes antagonized M3 mACHR activation at sub- to low micromolar concentrations, with UCL 1684 acting as an ASM relaxant. Caution should be taken when using these compounds to block SK potassium channels, as inhibition of mACHRs may be a side-effect if excessive concentrations are used.


Benefits of organic farming to biodiversity vary among taxa.
Habitat and biodiversity differences between matched pairs of organic and non-organic farms containing cereal crops in lowland England were assessed by a large-scale study of plants, invertebrates, birds and bats. Habitat extent, composition and management on organic farms was likely to favour higher levels of biodiversity and indeed organic farms tended to support higher numbers of species and overall abundance across most taxa. However, the magnitude of the response varied; plants showed larger and more consistent responses than other taxa. Variation in response across taxa may be partly a consequence of the small size and isolated context of many organic farms. Extension of organic farming could contribute to the restoration of biodiversity in agricultural landscapes.


Innovation institutions and the opioid crisis.
The US has recently-and belatedly-come to recognize opioid addiction as a public health crisis. What has gone mostly unrecognized is the degree to which this crisis is intertwined with US intellectual property law and related elements of US innovation policy. Innovation institutions-the legal arrangements that structure incentives for production and allocation of knowledge goods-encouraged the development and commercialization of addictive painkillers, restricted access to opioid antidotes, and (perhaps most importantly) failed to facilitate investments in alternative, nonaddictive treatments for chronic pain. Although innovation policy does not bear all the blame for the opioid wave that has washed over communities across the country, innovation institutions are bound up in the ongoing epidemic to a degree that so far has gone underappreciated. This article examines the proliferation of opioid use and abuse through the lens of innovation policy, and it envisions ways in which innovation institutions could help to contain the crisis. Along the way, it seeks to derive broader lessons for innovation policy scholarship as well as recommendations for institutional reform. The opioid crisis challenges the conventional understanding of IP law as a trade-off between allocative efficiency and dynamic efficiency; it highlights the potentially pernicious role of IP protection for addictive and habit-forming products; and it exposes deep flaws in the structure of federal subsidies for and regulation of prescription drugs. It also draws attention to the political and cultural factors that contribute to innovation policy failures. Ultimately, the opioid crisis underscores both the urgency and the limits of institutional change in the innovation policy domain.


Structural and functional analysis of raphe neurone implants into denervated rat spinal cord.
The ability of grafts of embryonic raphe cells to the adult rat spinal cord to reverse the morphological, neurochemical and functional deficits caused by ablation of the serotoninergic afferents has been studied. After grafting, extensive reinnervation was observed by 5HT-immunoreactive fibres, many of which appeared to make contacts with host motoneurones. The neurotransmitter complement was apparently normal. There was also a reinstatement of 5HT levels in the denervated cord after transplantation, amounting to some 40% of normal at the level of the graft. Similarly, Na+-dependent uptake of [3H]-5HT into P3 fractions was over 40% of that recorded in unlesioned animals. Antidromic stimulation of the ventral roots of the spinal cord was used to assess the degree of motoneurone excitability. The field potential in the ventral horn of grafted animals was increased by electrical stimulation of discrete regions along the cord, probably corresponding to the graft loci. It is concluded that serotoninergic neurones transplanted to the denervated spinal cord survive and develop normally, reinnervating the host tissue extensively. Furthermore, the graft/host connections appear to be functionally viable.


Daratumumab: A Review in Combination Therapy for Transplant-Eligible Newly Diagnosed Multiple Myeloma.
Intravenous daratumumab (DARZALEX®), a human monoclonal antibody targeting CD38, is approved in the EU and USA for use in combination with bortezomib, thalidomide and dexamethasone for the treatment of adults with newly diagnosed multiple myeloma (MM) who are eligible for autologous stem cell transplantation. A subcutaneous formulation of daratumumab has also been approved in the EU and USA (DARZALEX FASPRO™) for use in MM. In the pivotal phase III CASSIOPEIA trial in adults with newly diagnosed, transplant-eligible MM, the addition of intravenous daratumumab to bortezomib, thalidomide and dexamethasone significantly increased the proportion of patients with a stringent complete response and significantly prolonged progression-free survival; overall survival data are not yet mature. Some facets of health-related quality of life were improved by the addition of daratumumab. The addition of daratumumab had a minimal effect on overall toxicity and the most common grade ≥ 3 adverse events with daratumumab combination therapy were haematological (e.g. neutropenia, lymphopenia). The approval of daratumumab as combination therapy in patients with newly diagnosed, transplant-eligible MM expands the range of MM treatment settings in which daratumumab is an option and the availability of the subcutaneous formulation will likely be of benefit to patients.


Compact all-fiber high-energy fiber laser with sub-300-fs duration.
We report a compact all-fiber high-energy fiber laser that consists of a laser oscillator and a compression section. The laser oscillator generates the pulses with high energy and large chirp. The compression section is made of a piece of standard single-mode fiber that dechirps the chirped pulses. The compact all-fiber fiber laser produces pulses with 8 nJ of the pulse energy and 290 fs of the pulse duration.


Human-Based Advanced in vitro Approaches to Investigate Lung Fibrosis and Pulmonary Effects of COVID-19.
The coronavirus disease 2019 (COVID-19) pandemic has caused considerable socio-economic burden, which fueled the development of treatment strategies and vaccines at an unprecedented speed. However, our knowledge on disease recovery is sparse and concerns about long-term pulmonary impairments are increasing. Causing a broad spectrum of symptoms, COVID-19 can manifest as acute respiratory distress syndrome (ARDS) in the most severely affected patients. Notably, pulmonary infection with Severe Acute Respiratory Syndrome coronavirus 2 (SARS-CoV-2), the causing agent of COVID-19, induces diffuse alveolar damage (DAD) followed by fibrotic remodeling and persistent reduced oxygenation in some patients. It is currently not known whether tissue scaring fully resolves or progresses to interstitial pulmonary fibrosis. The most aggressive form of pulmonary fibrosis is idiopathic pulmonary fibrosis (IPF). IPF is a fatal disease that progressively destroys alveolar architecture by uncontrolled fibroblast proliferation and the deposition of collagen and extracellular matrix (ECM) proteins. It is assumed that micro-injuries to the alveolar epithelium may be induced by inhalation of micro-particles, pathophysiological mechanical stress or viral infections, which can result in abnormal wound healing response. However, the exact underlying causes and molecular mechanisms of lung fibrosis are poorly understood due to the limited availability of clinically relevant models. Recently, the emergence of SARS-CoV-2 with the urgent need to investigate its pathogenesis and address drug options, has led to the broad application of in vivo and in vitro models to study lung diseases. In particular, advanced in vitro models including precision-cut lung slices (PCLS), lung organoids, 3D in vitro tissues and lung-on-chip (LOC) models have been successfully employed for drug screens. In order to gain a deeper understanding of SARS-CoV-2 infection and ultimately alveolar tissue regeneration, it will be crucial to optimize the available models for SARS-CoV-2 infection in multicellular systems that recapitulate tissue regeneration and fibrotic remodeling. Current evidence for SARS-CoV-2 mediated pulmonary fibrosis and a selection of classical and novel lung models will be discussed in this review.


Plague in a Colony of Gunnison's Prairie Dogs ( Cynomys gunnisoni) Despite Three Years of Infusions of Burrows with 0.05% Deltamethrin to Kill Fleas.
At Valles Caldera National Preserve in New Mexico, US, infusing Gunnison's prairie dog ( Cynomys gunnisoni) burrows with an insecticide dust containing 0.05% deltamethrin killed fleas which transmit bubonic plague. The reduction in the number of fleas per prairie dog was significant and dramatic immediately after infusions, with a suggestion that the reduction persisted for as long as 12 mo. Despite the lower flea counts, however, a plague epizootic killed >95% of prairie dogs after 3 yr of infusions (once per year). More research is necessary for a better understanding of the efficacy of insecticide dusts at lowering flea counts and protecting prairie dogs from plague.


Toxicology and Carcinogenesis Studies of 4,4'-Diamino-2,2'-Stilbenedisulfonic Acid Disodium Salt (CAS No. 7336-20-1) in F344 Rats and B6C3F1 Mice (Feed Studies).
4,4'-Diamino-2,2'-stilbenedisulfonic acid, disodium salt, is used in the synthesis of dyes and optical brighteners or fluorescent whitening agents. Toxicology and carcinogenesis studies were conducted by administering the chemical (approximately 14% water, 6% sodium chloride, 4% impurities, and 76% 4,4'-diamino-2,2'-stilbenedisulfonic acid) in feed to groups of F344/N rats and B6C3F1 mice of each sex for 14 days, 13 weeks, and 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium and Chinese hamster ovary cells. 14-Day Studies: Groups of five rats and five mice of each sex were given 0, 6,250, 12,500, 25,000, 50,000, or 100,000 ppm 4,4'-diamino-2,2'-stilbenedisulfonic acid, disodium salt, in feed for 14 days. All rats and mice survived to the end of the studies. The mean body weight gain of male rats receiving 50,000 or 100,000 ppm and of female rats and male and female mice receiving 100,000 ppm was significantly lower than those of the respective controls. Clinical findings included diarrhea in the rats and mice receiving 100,000 ppm. There were no chemical-related changes in absolute or relative organ weights in rats or mice. There were no gross or microscopic lesions related to chemical administration in rats or mice. 13-Week Studies: Groups of 10 rats and 10 mice of each sex were given 0, 6,250, 12,500, 25,000, 50,000, or 100,000 ppm 4,4'-diamino-2,2'-stilbenedisulfonic acid, disodium salt, in feed for 13 weeks. One female rat, six male mice, and one female mouse in the 100,000 ppm dose groups died during the studies. Mean body weight gain was significantly decreased in male rats and female mice receiving 50,000 or 100,000 ppm, in male mice receiving 25,000, 50,000, or 100,000 ppm, and in female rats receiving 100,000 ppm. Clinical findings in rats that received 50,000 or 100,000 ppm and in mice that received 100,000 ppm included diarrhea, emaciation, and hyperemia of the perineum. There were no biologically significant changes in absolute or relative organ weights or clinical pathology results in rats or mice. Histopathologic lesions present in rats receiving 100,000 ppm were bone marrow hypercellularity and chronic inflammation of the anus and rectum. Ulcerative inflammation of the anus and rectum was observed in mice receiving 25,000 ppm and above. Female mice in the 6,250, 12,500, 25,000, and 50,000 ppm dose groups had increased incidences of cystic endometrial hyperplasia. 2-Year Studies: Doses selected for the 2-year studies were based on mortality, decreased body weight gains, and the presence of diarrhea and chronic inflammation of the anus/rectum in rats and mice during the 13-week studies. Groups of 60 rats of each sex were given 0, 12,500 or 25,000 ppm and groups of 60 mice of each sex were given 0, 6,250, or 12,500 ppm 4,4'-diamino-2,2'-stilbenedisulfonic acid, disodium salt, in feed for up to 103 weeks. Interim evaluations were performed on 10 rats and 10 mice from each dose group at 15 months. There were no biologically significant absolute or relative organ weight, clinical pathology, or histopathology findings in rats or mice administered 4,4'-diamino-2,2'-stilbenedisulfonic acid, disodium salt, in feed for 15 months. Body Weight, Feed Consumption, Survival, and Clinical Findings in the 2-Year Studies: Mean body weights were marginally decreased for high-dose male and female rats and female mice. Feed consumption by dosed rats and mice was similar to feed consumption by the controls throughout the studies. Survival was similar among control and treated groups of rats and mice. No clinical findings related to chemical administration were observed in rats or mice. Nonneoplastic and Neoplastic Effects in the 2-Year Studies: There were no chemical-related increased incidences of neoplasms at any site in rats. Ulcers of the forestomach or glandular stomach occurred in dosed rats (males: 1/50, 5/50, 4/50; females: 0/50, 1/50, 4/50), and may have been related to the administration of 4,4'-diamino-2,2'-stilbenedisulfonic acid, disodium salt. There were no chemical-related incidences ohemical-related incidences of neoplasms, nonneoplastic lesions, or other toxic effects in mice in the 2-year studies. Although the animals might have been able to tolerate slightly higher doses, results of the 13-week studies indicate that a doubling of the highest doses could not have been tolerated. Genetic Toxicology: 4,4'-Diamino-2,2'-stilbenedisulfonic acid was not mutagenic in Salmonella typhimurium strains TA100, TA1535, TA1537, or TA98 with or without S9 metabolic activation. 4,4'-Diamino-2,2'-stilbenedisulfonic acid did not induce sister chromatid exchanges or chromosomal aberrations in Chinese hamster ovary cells in the presence or absence of S9. Conclusions: Under the conditions of these 2-year feed studies, there was no evidence of carcinogenic activity of 4,4'-diamino-2,2'-stilbenedisulfonic acid, disodium salt, in male or female F344/N rats receiving 12,500 or 25,000 ppm. There was no evidence of carcinogenic activity of 4,4'-diamino-2,2'-stilbenedisulfonic acid, disodium salt, in male or female B6C3F1 mice receiving 6,250 or 12,500 ppm. Synonyms: Amsonic acid; diaminostilbene disulphonate (DASD); 2,2'-(1,2-ethenediyl)bis[5-amino-benzenesulfonic acid]; 2,2'-disulfo-4,4'-stilbenediamine; 2,2'-stilbenedisulfonic acid; 4,4'-diamino-2,2'-benzenesulfonic acid; 2,2'-(1,2-ethenediyl)bis(5-amino-) diaminostilbenedisulfonic acid; flavonic acid; p,p'-diaminostilbene-o,o'-disulfonic acid; 4,4'-diaminostilbene-2,2'-disulfonic acid


Alcohol screening and brief interventions in primary care - Evidence and a pragmatic practice-based approach.
Risky alcohol drinking is a common problem in adults presenting in Australian general practice. Preventive health guidelines recommend routine delivery of alcohol screening and brief intervention (ASBI) by general practitioners (GPs). However, ASBIs have rarely been implemented successfully in a sustainable manner. In this article, we explain the current state of empirical evidence for the effectiveness of ASBI in primary care and describe a pragmatic interpretation of how this evidence applies to routine care. The empirical evidence surrounding ASBIs is complex. ASBIs are efficacious in research settings, but their effectiveness when compared with control interventions in real-world practice is less certain. Alcohol assessment within therapeutic doctor-patient relationships, rather than the specific formal tools, may be the 'active ingredient'. A pragmatic, practice-based approach to early detection of risky drinking is to focus on strategies that allow asking patients about their drinking more regularly, documenting it, and using quality improvement methodology to improve alcohol recording data completeness for the practice population.


Decrease in heart rate after longitudinal participation in the Groningen Active Living Model (GALM) recreational sports programme.
The aim of this study was to investigate changes in heart rate during submaximal exercise as an index of cardiovascular function in older adults participating in the Groningen Active Living Model recreational sports programme who were sedentary or underactive at baseline. A repeated measurement design was conducted; 151 participants were included, providing 398 heart rate files over a period of 18 months. Multi-level analyses were conducted; growth and final models were developed. Significant decreases in mean heart rate over time were observed for all walking speeds. The covariates of sex and body mass index (BMI) were significantly related to mean heart rate at each walking speed, except for BMI at 7 km x h(-1). No significant relationships were observed between energy expenditure for recreational sports activities and leisure-time physical activities and mean heart rate, except for energy expenditure for leisure-time physical activities at 7 km x h(-1). From baseline to December 2002, decreases in predicted mean heart rate were 5.5, 6.0, 10.0, and 9.0 beats x min(-1) at walking speeds of 4, 5, 6, and 7 km x h(-1); relative decreases ranged from 5.1 to 7.4%. Significant decreases in heart rate observed during submaximal exercise reflected a potential increase in cardiovascular function after 18 months of participation in the Groningen Active Living Model recreational sports programme.


Efficiency and specificity of microRNA-primed nucleotide analog incorporation by various DNA polymerases.
MicroRNAs (miRNAs) as endogenous regulators of gene expression have spurred a surge of interest for their quantification and expression analysis. High-sensitivity and high-specificity miRNA detection techniques, such as real-time polymerase chain reaction and recently introduced bioluminescent miRNA detection, require systematic study of DNA polymerases for use with miRNAs. In this study, a variety of DNA polymerases were studied to assess their capabilities of using miRNA as a primer and incorporating 2'-deoxyadenosine-5'-O-(1-thiotriphosphate) as a dATP alternative during DNA strand extension. Five DNA polymerases were investigated: mesophilic DNA polymerase I large (Klenow) fragment, 3'-->5' exo(-) Klenow DNA polymerase, thermophilic Bst DNA polymerase large fragment, Therminator DNA polymerase, and Taq DNA polymerase. The experimental results show that, except for Taq DNA polymerase, the polymerases can use miRNA as a primer and have both common and divergent properties of the nucleotide analog incorporation and miRNA discrimination. DNA polymerase I large (Klenow) fragment showed no detectable polymerization product with the thio-modified dATP as a substrate. Thermophilic Bst DNA polymerase had the highest specificity for miRNA recognition on a DNA template. The study provides a novel method for miRNA detection without reverse transcription to complementary DNA that is faster, simpler, and less prone to biases and errors.


Intrathecal baclofen for management of spastic cerebral palsy: multicenter trial.
Intrathecal baclofen infusion has demonstrated effectiveness in decreasing spasticity of spinal origin. Oral antispasticity medication is minimally effective or not well tolerated in cerebral palsy. This study assessed the effectiveness of intrathecal baclofen in reducing spasticity in cerebral palsy. Candidates were screened by randomized, double-blind, intrathecal injections of baclofen and placebo. Responders were defined as those who experienced an average reduction of 1.0 in the lower extremities on the Ashworth Scale for spasticity. Responders received intrathecal baclofen via the SynchroMed System and were followed for up to 43 months. Fifty-one patients completed screening and 44 entered open-label trials. Lower-extremity spasticity decreased from an average baseline score of 3.64 to 1.90 at 39 months. A decrease in upper extremity spasticity was evidenced over the same study period. Forty-two patients reported adverse events. Most common reports were hypotonia, seizures (no new onset), somnolence, and nausea or vomiting. Fifty-nine percent of the patients experienced procedural or system-related events. Spasticity in patients with cerebral palsy can be treated effectively by continuous intrathecal baclofen. Adverse events, although common, were manageable.


Nanografting de novo proteins onto gold surfaces.
The immobilization of novel proteins onto addressable locations on a flat surface has potential applications in a range of biotechnologies. Here we describe the nanopatterning of a de novo protein onto a gold surface. Patterning was achieved using a technique called nanografting, in which the tip of an atomic force microscope is used to disrupt a preexisting monolayer of alkanethiol molecules on a gold surface, thereby facilitating exchange with alternative thiol-linked molecules from the surrounding solution. The protein used for these studies was chosen from a designed combinatorial library of de novo sequences expressed in E. coli and was engineered to have a glycine-glycine-cysteine tag at its C-terminus, thereby enabling attachment to the gold surface through a single cysteine thiol. The average height of the grafted protein patterns was found to be somewhat higher than expected from the known NMR structure of the protein. Compression of the nanografted patches by an external force (below 10 nN) was reversible but showed some hysteresis. Interestingly, both the energy required to deform the immobilized protein patterns and the energy defined by the hysteresis loop were found to be of the same order as the energy required to unfold the monomeric protein in solution. These studies demonstrate the possibility of preparing nanometer scale protein arrays, lowering significantly the volume requirements of the protein samples necessary to fabricate protein-based biosensor arrays and thereby providing a base for increasing their sensitivity.


Xylem ionic relations and salinity tolerance in barley.
Control of ion loading into the xylem has been repeatedly named as a crucial factor determining plant salt tolerance. In this study we further investigate this issue by applying a range of biophysical [the microelectrode ion flux measurement (MIFE) technique for non-invasive ion flux measurements, the patch clamp technique, membrane potential measurements] and physiological (xylem sap and tissue nutrient analysis, photosynthetic characteristics, stomatal conductance) techniques to barley varieties contrasting in their salt tolerance. We report that restricting Na(+) loading into the xylem is not essential for conferring salinity tolerance in barley, with tolerant varieties showing xylem Na(+) concentrations at least as high as those of sensitive ones. At the same time, tolerant genotypes are capable of maintaining higher xylem K(+)/Na(+) ratios and efficiently sequester the accumulated Na(+) in leaves. The former is achieved by more efficient loading of K(+) into the xylem. We argue that the observed increases in xylem K(+) and Na(+) concentrations in tolerant genotypes are required for efficient osmotic adjustment, needed to support leaf expansion growth. We also provide evidence that K(+)-permeable voltage-sensitive channels are involved in xylem loading and operate in a feedback manner to maintain a constant K(+)/Na(+) ratio in the xylem sap.


Combined vitamin B6-magnesium treatment in autism spectrum disorder.
The use of mega-vitamin intervention began in the early 1950's with the treatment of schizophrenic patients. Pyroxidine (vitamin B6) was first used with children diagnosed with "autism syndrome" when speech and language improvement was observed in some children as a result of large doses of B6. A number of published studies attempted to assess the effects of vitamin B6-Mg (Mg was found to reduce undesirable side effects from B6) on a variety of characteristics such as verbal communication, non-verbal communication, interpersonal skills, and physiological function, in individuals with autism. To determine the efficacy of vitamin B6 and magnesium (B6-Mg) for treating social, communication and behavioural responses of children and adults with autism. We searched the Cochrane Controlled Trials Register (Cochrane Library, Issue 2, 2002), MEDLINE (1966- January 2002), EMBASE (1980-January 2002), PsychINFO (1887 - January 2002), Dissertation Abstracts International (1861 - January 2002). The search engine FirstSearch was also used (January 2002). Reference lists for all the obtained studies and other review articles were examined for additional studies. All studies in which the participants were randomly allocated prior to intervention and in which outcomes were compared to either a placebo or non-treated group were included. Two reviewers independently evaluated all potential studies identified as indicated above for inclusion. Two trials were included in the review. Both studies used a double-blind crossover design. One study (Tolbert 1993) provided insufficient data to conduct an analysis. The senior author was contacted for supporting data but was unable to provide the needed information. The remaining study (Findling, 1997) yielded no significant differences between treatment and placebo group performances following the B6 intervention on measures of social interaction, communication, compulsivity, impulsivity, or hyperactivity. Due to the small number of studies, the methodological quality of studies, and small sample sizes, no recommendation can be advanced regarding the use of B6-Mg as a treatment for autism.


Efficacy of an Internet and SMS-based integrated smoking cessation and alcohol intervention for smoking cessation in young people: study protocol of a two-arm cluster randomised controlled trial.
Tobacco smoking prevalence continues to be high, particularly among adolescents and young adults with lower educational levels, and is therefore a serious public health problem. Tobacco smoking and problem drinking often co-occur and relapses after successful smoking cessation are often associated with alcohol use. This study aims at testing the efficacy of an integrated smoking cessation and alcohol intervention by comparing it to a smoking cessation only intervention for young people, delivered via the Internet and mobile phone. A two-arm cluster-randomised controlled trial with one follow-up assessment after 6 months will be conducted. Participants in the integrated intervention group will: (1) receive individually tailored web-based feedback on their drinking behaviour based on age and gender norms, (2) receive individually tailored mobile phone text messages to promote drinking within low-risk limits over a 3-month period, (3) receive individually tailored mobile phone text messages to support smoking cessation for 3 months, and (4) be offered the option of registering for a more intensive program that provides strategies for smoking cessation centred around a self-defined quit date. Participants in the smoking cessation only intervention group will only receive components (3) and (4). Study participants will be 1350 students who smoke tobacco daily/occasionally, from vocational schools in Switzerland. Main outcome criteria are 7-day point prevalence smoking abstinence and cigarette consumption assessed at the 6-month follow up. This is the first study testing a fully automated intervention for smoking cessation that simultaneously addresses alcohol use and interrelations between tobacco and alcohol use. The integrated intervention can be easily implemented in various settings and could be used with large groups of young people in a cost-effective way. Current Controlled Trials ISRCTN02427446 (date of registration: 08th September, 2014).


Definition of perforator flap: what does a "perforator" perforate?
Perforator flap concept plays an important role in reconstructive surgery, because it allows less invasive and more complex reconstruction by preserving major vessels and muscles with intramuscular vessel dissection. Originally "perforator" represents vessel perforating the muscle, then vessel perforating the deep fascia regardless of muscle perforation. With technical progress in reconstructive microsurgery, the previous definition becomes inappropriate for least invasive flaps, only requiring intra-adiposal vessel dissection, such as superficial circumflex iliac artery perforator flap. Based on our experience of various least invasive flap reconstructive surgeries, a new concept for perforator flap has been developed. The new definition of perforator is a vessel perforating an envelope of a targeted tissue to be transferred; the superficial fascia for skin, the periosteum for bone, the perineurium for nerve, and the deep fascia for muscle. According to the new definition, all flaps can be precisely classified based on the corresponding "perforator".


Assays of urine levels of HNL/NGAL in patients undergoing cardiac surgery and the impact of antibody configuration on their clinical performances.
Acute kidney injury (AKI) is one of the most serious postoperative complications of cardiac surgery. The lack of early and powerful markers for AKI makes the morbidity and mortality still very high. HNL (Human neutrophil lipocalin)/NGAL (Neutrophil gelatinase-associated lipocalin) was recently shown as a novel biomarker for AKI after cardiac surgery. Serial urine samples from 59 patients undergoing cardiac surgery were analyzed by polyclonal antibody based radioimmunoassay (RIA), monoclonal-polyclonal antibody based enzyme-linked immunosorbent assay (ELISA). We found 10 to 100-fold increases in urine HNL/NGAL levels in about half of the patients 2 h after termination of the operation and elevated levels in all patients 72 h post operation. The urine levels of HNL/NGAL showed a weak, but significant relation with kidney function as measured by plasma levels of cystatin C or creatinine. The 2 h-HNL/NGAL levels were positively correlated to extracorporeal circulation time (p<0001). The assays were well correlated, but had different clinical performances. We confirmed that urine HNL/NGAL may be a useful early biomarker of postoperative kidney injury. The results indicate that the antibody configuration of the assay has an impact on the clinical performance of the assay.


Chemicals having estrogenic activity can be released from some bisphenol A-free, hard and clear, thermoplastic resins.
Chemicals that have estrogenic activity (EA) can potentially cause adverse health effects in mammals including humans, sometimes at low doses in fetal through juvenile stages with effects detected in adults. Polycarbonate (PC) thermoplastic resins made from bisphenol A (BPA), a chemical that has EA, are now often avoided in products used by babies. Other BPA-free thermoplastic resins, some hypothesized or advertised to be EA-free, are replacing PC resins used to make reusable hard and clear thermoplastic products such as baby bottles. We used two very sensitive and accurate in vitro assays (MCF-7 and BG1Luc human cell lines) to quantify the EA of chemicals leached into ethanol or water/saline extracts of fourteen unstressed or stressed (autoclaving, microwaving, UV radiation) thermoplastic resins. Estrogen receptor (ER)-dependent agonist responses were confirmed by their inhibition with the ER antagonist ICI 182,780. Our data showed that some (4/14) unstressed and stressed BPA-free thermoplastic resins leached chemicals having significant levels of EA, including one polystyrene (PS), and three Tritan™ resins, the latter reportedly EA-free. Exposure to UV radiation in natural sunlight resulted in an increased release of EA from Tritan™ resins. Triphenyl-phosphate (TPP), an additive used to manufacture some thermoplastic resins such as Tritan™, exhibited EA in both MCF-7 and BG1Luc assays. Ten unstressed or stressed glycol-modified polyethylene terephthalate (PETG), cyclic olefin polymer (COP) or copolymer (COC) thermoplastic resins did not release chemicals with detectable EA under any test condition. This hazard survey study assessed the release of chemicals exhibiting EA as detected by two sensitive, widely used and accepted, human cell line in vitro assays. Four PC replacement resins (Tritan™ and PS) released chemicals having EA. However, ten other PC-replacement resins did not leach chemicals having EA (EA-free-resins). These results indicate that PC-replacement plastic products could be made from EA-free resins (if appropriate EA-free additives are chosen) that maintain advantages of re-usable plastic items (price, weight, shatter resistance) without releasing chemicals having EA that potentially produce adverse health effects on current or future generations.


Do dual-mobility cups cemented into porous tantalum shells reduce the risk of dislocation after revision surgery?
Background and purpose - Dual-mobility cups (DMCs) reduce the risk of dislocation and porous tantalum (TM) shells show favorable osseointegration after acetabular revision surgery, yet the combination of these implants has not been studied. We hypothesized that (1) cementing a DMC into a TM shell decreases the risk of dislocation; (2) DMCs cemented into TM shells are not at greater risk of re-revision; (3) liberation of tantalum ions is marginal after use of this combined technique. Patients and methods - We investigated the outcome in 184 hips (184 patients) after acetabular revision surgery with TM shells, fitted either with DMCs (n = 69), or with standard poly-ethylene (PE) liners (n = 115). Chart follow-up was complete for all patients, and the occurrence of dislocations and re-revisions was recorded. 20 were deceased, 50 were unable to attend follow-up, leaving 114 for assessment of hip function after 4.9 (0.5-8.9) years, radiographs were obtained in 99, and tantalum concentrations in 84 patients. Results - 1 patient with a DMC had a dislocation, whereas 14 patients with PE liners experienced at least 1 dislocation. 11 of 15 re-revisions in the PE group were necessitated by dislocations, whereas none of the 2 re-revisions in the DMC group was performed for this reason. Hence, dislocation-free survival after 4 years was 99% (95% CI 96-100) in the DMC group, whereas it was 88% (CI 82-94, p = 0.01) in the PE group. We found no radiographic signs of implant failure in any patient. Mean tantalum concentrations were 0.1 µl/L (CI 0.05-0.2) in the DMC group and 0.1 µg/L (CI 0.05-0.2) in the PE group. Interpretation - Cementing DMCs into TM shells reduces the risk of dislocation after acetabular revision surgery without jeopardizing overall cup survival, and without enhancing tantalum release.


Consequences of transverse acetabular fracture malreduction on load transmission across the hip joint.
To evaluate the biomechanical behavior of gap and step malreductions in a model of transverse acetabular fracture. Cadaver pelvis loading in simulated single-leg stance with intact acetabulum, after transverse acetabular fracture anatomically reduced, and after step and gap malreduction. Five transtectal transverse fractures; five juxtatectal transverse fractures. Quasi-static loading of the hip with simulated abductor mechanism to physiologic loads with pressure-sensitive film interposed in the joint to determine contact area and contact pressure within the hip joint. Hip joint contact parameters: contact area, peak and mean contact pressure, and load distribution. Step malreduction of the transtectal transverse fracture resulted in significantly increased peak contact pressures (20.5 megapascals) in the superior acetabular articular surface as opposed to the intact acetabulum (9.1 megapascals). Gap malreduction of transtectal transverse fracture and step and gap malreduction of juxtatectal fracture did not result in significantly increased contact pressures in the hip. Step malreduction of a transverse acetabular fracture in the superior articular surface results in abnormally high contact forces and may predispose to the development of posttraumatic arthritis.


Dendritic cell therapy of high-grade gliomas.
The prognosis of patients with malignant glioma is poor in spite of multimodal treatment approaches consisting of neurosurgery, radiochemotherapy and maintenance chemotherapy. Among innovative treatment strategies like targeted therapy, antiangiogenesis and gene therapy approaches, immunotherapy emerges as a meaningful and feasible treatment approach for inducing long-term survival in at least a subpopulation of these patients. Setting up immunotherapy for an inherent immunosuppressive tumor located in an immune-privileged environment requires integration of a lot of scientific input and knowledge of both tumor immunology and neuro-oncology. The field of immunotherapy is moving into the direction of active specific immunotherapy using autologous dendritic cells (DCs) as vehicle for immunization. In the translational research program of the authors, the whole cascade from bench to bed to bench of active specific immunotherapy for malignant glioma is covered, including proof of principle experiments to demonstrate immunogenicity of patient-derived mature DCs loaded with autologous tumor lysate, preclinical in vivo experiments in a murine orthotopic glioma model, early phase I/II clinical trials for relapsing patients, a phase II trial for patients with newly diagnosed glioblastoma (GBM) for whom immunotherapy is integrated in the current multimodal treatment, and laboratory analyses of patient samples. The strategies and results of this program are discussed in the light of the internationally available scientific literature in this fast-moving field of basic science and translational clinical research.


Pharmacy Interweaving Safety Within Hospital Health Information Technology.
Research on the impact of hospital technology on medication safety usually focuses on prescribing and administration. Less is known about the pharmacy-related processes of reviewing, ordering and dispensing medications and how technology supports this work. We carried out a qualitative exploratory study of a hospital in England (UK) with the aim of gaining insight into processes of digitalisation. We found that hospital pharmacy staff perform safety work with technology aiming to prevent harm, such as 'scaffolding' people's thinking processes, or linking-up unintegrated systems. Their work seems to 'interweave' safety in between others' medication activities, but they do so sometimes struggling with technological deficiencies.


Overturning refusal of a hospital to terminate life support for a brain-dead mother until the fetus was born: What is the law in South Africa?
In a Texas case the court granted a husband an order for the removal of life support from his brain-dead pregnant wife after a hospital tried to keep her on it until the fetus was born. In South Africa the court would have issued a similar order, but for different reasons. Here, unlawfully and intentionally subjecting a pregnant corpse to life-support measures to keep a fetus alive against the wishes of the family would amount to the crime of violating a corpse.


Influences of laser source on phase-sensitivity optical time-domain reflectometer-based distributed intrusion sensor.
This paper investigates the influences of laser source on distributed intrusion sensor based on a phase-sensitivity optical time-domain reflectometer (φ-OTDR). A numerical simulation is performed to illustrate the relationships between trace-to-trace fluctuations and frequency drift rate as well as pulse width, and fluctuations ratio coefficient (FRC) is proposed to evaluate the level of trace-to-trace fluctuations. The simulation results show that the FRC grows with increasing frequency drift rate and pulse width, reaches, and maintains the peak value when the frequency drift rate and/or the pulse width are high enough. Furthermore, experiments are implemented using a φ-OTDR prototype with a low frequency drift laser (<5 MHz/min), of which the high frequency drift rate is simulated by frequency sweeping. The good agreement of experimental with simulated results in the region of high frequency drift rate validates the theoretical analysis, and the huge differences between them in the region of low frequency drift rate indicate the place of laser frequency drift among system noises. The conclusion is useful for choosing laser sources and improving the performance of φ-OTDR.


Delayed dynamical systems: networks, chimeras and reservoir computing.
We present a systematic approach to reveal the correspondence between time delay dynamics and networks of coupled oscillators. After early demonstrations of the usefulness of spatio-temporal representations of time-delay system dynamics, extensive research on optoelectronic feedback loops has revealed their immense potential for realizing complex system dynamics such as chimeras in rings of coupled oscillators and applications to reservoir computing. Delayed dynamical systems have been enriched in recent years through the application of digital signal processing techniques. Very recently, we have showed that one can significantly extend the capabilities and implement networks with arbitrary topologies through the use of field programmable gate arrays. This architecture allows the design of appropriate filters and multiple time delays, and greatly extends the possibilities for exploring synchronization patterns in arbitrary network topologies. This has enabled us to explore complex dynamics on networks with nodes that can be perfectly identical, introduce parameter heterogeneities and multiple time delays, as well as change network topologies to control the formation and evolution of patterns of synchrony. This article is part of the theme issue 'Nonlinear dynamics of delay systems'.


SGLT2 Inhibitors as Calorie Restriction Mimetics: Insights on Longevity Pathways and Age-Related Diseases.
Sodium-glucose cotransporter-2 (SGLT2) inhibitors induce glycosuria, reduce insulin levels, and promote fatty acid oxidation and ketogenesis. By promoting a nutrient deprivation state, SGLT2 inhibitors upregulate the energy deprivation sensors AMPK and SIRT1, inhibit the nutrient sensors mTOR and insulin/IGF1, and modulate the closely linked hypoxia-inducible factor (HIF)-2α/HIF-1α pathways. Phosphorylation of AMPK and upregulation of adiponectin and PPAR-α favor a reversal of the metabolic syndrome which have been linked to suppression of chronic inflammation. Downregulation of insulin/IGF1 pathways and mTOR signaling from a reduction in glucose and circulating amino acids promote cellular repair mechanisms, including autophagy and proteostasis which confer cellular stress resistance and attenuate cellular senescence. SIRT1, another energy sensor activated by NAD+ in nutrient-deficient states, is reciprocally activated by AMPK, and can deacetylate and activate transcription factors, such as PCG-1α, mitochondrial transcription factor A (TFAM), and nuclear factor E2-related factor (NRF)-2, that regulate mitochondrial biogenesis. FOXO3 transcription factor which target genes in stress resistance, is also activated by AMPK and SIRT1. Modulation of these pathways by SGLT2 inhibitors have been shown to alleviate metabolic diseases, attenuate vascular inflammation and arterial stiffness, improve mitochondrial function and reduce oxidative stress-induced tissue damage. Compared with other calorie restriction mimetics such as metformin, rapamycin, resveratrol, and NAD+ precursors, SGLT2 inhibitors appear to be the most promising in the treatment of aging-related diseases, due to their regulation of multiple longevity pathways that closely resembles that achieved by calorie restriction and their established efficacy in reducing cardiovascular events and all-cause mortality. Evidence is compelling for the role of SGLT2 inhibitors as a calorie restriction mimetic in anti-aging therapeutics.


The role of conformation on electron capture dissociation of ubiquitin.
Effects of protein conformation on electron capture dissociation (ECD) were investigated using high-field asymmetric waveform ion mobility spectrometry (FAIMS) and Fourier-transform ion cyclotron resonance mass spectrometry. Under the conditions of these experiments, the electron capture efficiency of ubiquitin 6+ formed from three different solution compositions differs significantly, ranging from 51 +/- 7% for ions formed from an acidified water/methanol solution to 88 +/- 2% for ions formed from a buffered aqueous solution. This result clearly indicates that these protein ions retain a memory of their solution-phase structure and that conformational differences can be probed in an ECD experiment. Multiple conformers for the 7+ and 8+ charge states of ubiquitin were separated using FAIMS. ECD spectra of conformer selected ions of the same charge states differ both in electron capture efficiency and in the fragment ion intensities. Conformers of a given charge state that have smaller collisional cross sections can have either a larger or smaller electron capture efficiency. A greater electron capture efficiency was observed for ubiquitin 6+ that has the same collisional cross section as one ubiquitin 7+ conformer, despite the lower charge state. These results indicate that the shape of the molecule can have a greater effect on electron capture efficiency than either collisional cross section or charge state alone. The cleavage locations of different conformers of a given charge state were the same indicating that the presence of different conformers in the gas phase is not due to difference in where charges are located, but rather reflect conformational differences most likely originating from solution. Small neutral losses observed from the singly- and doubly-reduced ubiquitin 6+ do not show a temperature dependence to their formation, consistent with these ions being formed by nonergodic processes.


Protease-activated receptor-2 activation exaggerates TRPV1-mediated cough in guinea pigs.
A lowered threshold to the cough response frequently accompanies chronic airway inflammatory conditions. However, the mechanism(s) that from chronic inflammation results in a lowered cough threshold is poorly understood. Irritant agents, including capsaicin, resiniferatoxin, and citric acid, elicit cough in humans and in experimental animals through the activation of the transient receptor potential vanilloid 1 (TRPV1). Protease-activated receptor-2 (PAR2) activation plays a role in inflammation and sensitizes TRPV1 in cultured sensory neurons by a PKC-dependent pathway. Here, we have investigated whether PAR2 activation exaggerates TRPV1-dependent cough in guinea pigs and whether protein kinases are involved in the PAR2-induced cough modulation. Aerosolized PAR2 agonists (PAR2-activating peptide and trypsin) did not produce any cough per se. However, they potentiated citric acid- and resiniferatoxin-induced cough, an effect that was completely prevented by the TRPV1 receptor antagonist capsazepine. In contrast, cough induced by hypertonic saline, a stimulus that provokes cough in a TRPV1-independent manner, was not modified by aerosolized PAR2 agonists. The PKC inhibitor GF-109203X, the PKA inhibitor H-89, and the cyclooxygenase inhibitor indomethacin did not affect cough induced by TRPV1 agonists, but abated the exaggeration of this response produced by PAR2 agonists. In conclusion, PAR2 stimulation exaggerates TRPV1-dependent cough by activation of diverse mechanism(s), including PKC, PKA, and prostanoid release. PAR2 activation, by sensitizing TRPV1 in primary sensory neurons, may play a role in the exaggerated cough observed in certain airways inflammatory diseases such as asthma and chronic obstructive pulmonary disease.


Characterization and functional expression of a rat genomic DNA clone encoding a lymphocyte potassium channel.
Low stringency hybridization screening of a rat genomic DNA library with a previously described cDNA clone encoding a rat voltage-gated potassium channel has resulted in the characterization of a member of the potassium channel family, RGK5. An uninterrupted nucleotide sequence encodes a protein 525 amino acids in length, revealing that the entire coding region resides on a single exon. RGK5 transcripts are present in both mouse thymus and rat brain, as determined by Northern blot analysis. RNA transcribed in vitro from RGK5 genomic DNA directs the expression of functional potassium currents after injection into Xenopus oocytes. The currents are activated by depolarization, being half-activated at -14 mV, and inactivate almost completely during depolarizations of 1 to 2 s. The properties of the currents strongly resemble those of the type n potassium channel present on both immature thymocytes and Th lymphocytes.


Resonance Raman investigation of the interaction of thromboxane synthase with substrate analogues.
Thromboxane synthase is a hemethiolate enzyme that catalyzes the isomerization of prostaglandin H2 to thromboxane A2. We report the first resonance Raman (RR) spectra of recombinant human thromboxane synthase (TXAS) in both the presence and the absence of substrate analogues U44069 and U46619. The resting enzyme and its U44069 complex are found to have a 6-coordinate, low spin (6c/ls) heme, in agreement with earlier experiments. The U46619-bound enzyme is detected as a 6c/ls heme too, which is in contradiction with a previous conclusion based on absorption difference spectroscopy. Two new vibrations at 368 and 424 cm(-1) are observed upon binding of the substrate analogues in the heme pocket and are assigned to the second propionate and vinyl bending modes, respectively. We interpret the changes in these vibrational modes as the disruption of the protein environment and the hydrogen-bonding network of one of the propionate groups when the substrate analogues enter the heme pocket. We use carbocyclic thromboxane A2 (CTA2) to convert the TXAS heme cofactor to its 5-coordinate, high spin (5c/hs) form, as is confirmed by optical and RR spectroscopy. In this 5c/hs state of the enzyme, the Fe-S stretching frequency is determined at 350 cm(-1) with excitation at 356.4 nm. This assignment is supported by comparison to the spectrum of resting enzyme excited at 356.4 nm and by exciting at different wavelengths. Implications of our findings for substrate binding and the catalytic mechanism of TXAS will be discussed.


Self-managed programmes in homeless care as (reinvented) institutions.
Purpose: Self-managed institutional homeless programmes started as an alternative to regular shelters. Using institutional theory as a lens, we aim to explore the experiences of stakeholders with the institutional aspects of a self-managed programs.Method: The data we analysed (56 interviews, both open and semi-structured) were generated in a longitudinal participatory case-study into JES, a self-managed homeless shelter. In our analysis we went back and forth between our empirical data and theory, using a combination of systematic coding and interpretation. Participants were involved in all stages of the research.Results: Our analysis revealed similarities between JES and regular shelters, stemming from institutional similarities. Participants shared space and facilities with sixteen people, which caused an ongoing discussion on (enforcement of) rules. Participants loathed lack of private space. However, participants experienced freedom of choice over both their own life and management of JES and structures were experienced more fluid than in regular care. Some structures also appeared stimulated self-management.Conclusion: Our analysis showed how an institutional context influences self-management and suggested opportunities for introducing freedom and fluidity in institutional care.


Clinical profile and course of children with postinfectious bronchiolitis obliterans from a tertiary care hospital.
Postinfectious bronchiolitis obliterans (PIBO) is a chronic obstructive lung disease with scanty information in literature on etiology, clinical profile, treatment, and outcome. The objective of the study is to describe the clinical profile and course of children diagnosed with PIBO. A chart review of children below 18 years of age diagnosed as PIBO over the past 9 years was carried out. Details of clinical profile, laboratory investigations, imaging, treatment received, and outcome were recorded. Eight children (boys 4) with PIBO were identified. Median (interquartile range [IQR]) age at the first episode of acute severe bronchiolitis such as illness and diagnosis of PIBO was 15 (6, 23.5) and 30 (16.5, 60) months, respectively, indicating a delay in diagnosis. The most common symptoms were recurrent episodes of cough (100%), fast breathing (100%), wheezing (87.5%), and fever (62.5%). Median (IQR) number of hospitalizations and episodes of antibiotic use prior to diagnosis were 2.5 (2, 5.5) and 2 (2, 4), respectively. Three (37.5%) children received mechanical ventilation during previous hospitalizations. Chest computed tomography revealed mosaic attenuation in 8 (100%), ground-glass opacities in 2 (25%), and bronchial wall thickening in 2 (25%). After diagnosis, 7 received oral steroids, 7 received hydroxychloroquine, 5 received azithromycin, and 2 received azathioprine. The median (IQR) duration of follow-up (n = 6) was 6 (1.5, 9.5) months. Median (IQR) number of pulmonary exacerbations in follow-up was 2 (1, 5). PIBO is still an under-recognized entity with substantial delay in diagnosis and unnecessary use of antibiotics. Clinical course with imaging findings may help to diagnose and manage this entity.


Single-Agent Mosunetuzumab Shows Durable Complete Responses in Patients With Relapsed or Refractory B-Cell Lymphomas: Phase I Dose-Escalation Study.
Mosunetuzumab is a bispecific antibody targeting CD20 and CD3 that redirects T cells to engage and eliminate malignant B cells and is being developed for relapsed or refractory (R/R) B-cell non-Hodgkin lymphomas (B-NHLs). This first-in-human trial (ClinicalTrials.gov identifier: NCT02500407) evaluated the safety and tolerability and efficacy of mosunetuzumab in patients with R/R B-NHL and established the recommended phase II dose. Data from dose escalation are presented. Single-agent mosunetuzumab was administered intravenously in 3-week cycles, at full dose in cycle 1 day 1 (group A) or with ascending (step-up) doses during cycle 1 on days 1, 8, and 15 (group B), for eight or 17 cycles on the basis of tumor response. Two hundred thirty patients were enrolled. Doses up to 2.8 mg and 60 mg were assessed in groups A and B, respectively; maximum tolerated dose was not exceeded. In group B (n = 197), common adverse events (≥ 20% of patients) were neutropenia (28.4%), cytokine release syndrome (27.4%), hypophosphatemia (23.4%), fatigue (22.8%), and diarrhea (21.8%). Cytokine release syndrome was mostly low-grade (grade ≥ 3: 1.0%) and mainly confined to cycle 1. Across the doses investigated (group B), best overall response rates were 34.9% and 66.2% in patients with aggressive and indolent B-NHL, respectively, and complete response rates were 19.4% and 48.5%. Among patients with a complete response, the median duration of response was 22.8 months (95% CI, 7.6 to not estimable) and 20.4 (95% CI, 16 to not estimable) in patients with aggressive and indolent B-NHL, respectively. Mosunetuzumab, administered with step-up dosing, has a manageable safety profile and induces durable complete responses in R/R B-NHL. The expansion stage of the study is ongoing at the dose level of 1/2/60/60/30 mg selected for further study.


Oral 28-day and developmental toxicity studies of (R)-3-hydroxybutyl (R)-3-hydroxybutyrate.
(R)-3-Hydroxybutyl (R)-3-hydroxybutyrate (ketone monoester) has been developed as an oral source of ketones, which may be utilized for energy. In a 28-day toxicity study, Crl:WI (Wistar) rats received diets containing, as 30% of the calories, ketone monoester (12 and 15 g/kg body weight/day for male and female rats, respectively). Control groups received either carbohydrate- or fat-based diets. Rats in the test group consumed less feed and gained less weight than control animals; similar findings have been documented in studies of ketogenic diets. Between-group differences were noted in selected hematology, coagulation, and serum chemistry parameters; however, values were within normal physiological ranges and/or were not accompanied by other changes indicative of toxicity. Upon gross and microscopic evaluation, there were no findings associated with the ketone monoester. In a developmental toxicity study, pregnant Crl:WI (Han) rats were administered 2g/kg body weight/day ketone monoester or water (control) via gavage on days 6 through 20 of gestation. No Caesarean-sectioning or litter parameters were affected by the test article. The overall incidence of fetal alterations was higher in the test group; however, there were no specific alterations attributable to the test substance. The results of these studies support the safety of ketone monoester.


Overdiagnosis and overtreatment of thyroid cancer: A population-based temporal trend study.
The increase in incidence of thyroid cancer during the last decades without concomitant rise in mortality may reflect the growing detection of indolent forms of thyroid cancer, and may have fueled unnecessary thyroidectomies. Our aim was therefore, to compare recent secular trends in surgical intervention rate for thyroid cancer with the incidence and mortality of thyroid cancer to assess overdiagnosis and resulting overtreatment. We conducted a population-based temporal trend study in Switzerland from 1998 to 2012. All cases of invasive thyroid cancer, deaths from thyroid cancer, and cancer-related thyroidectomies were analyzed. We calculated changes in age-standardized thyroid cancer incidence rates, stratified by histologic subtype and tumor stage, thyroid cancer-specific mortality, and thyroidectomy rates. Between 1998 and 2012, the age-standardized annual incidence of thyroid cancer increased from 5.9 to 11.7 cases/100,000 among women (annual mean absolute increase: +0.43/100,000/year) and from 2.7 to 3.9 cases/100,000 among men (+0.11/100,000/year). The increase was limited to the papillary subtype, the most indolent form of thyroid cancer. The incidence of early stages increased sharply, the incidence of advanced stages increased marginally, and the mortality from thyroid cancer decreased slightly. There was a three- to four-fold increase in the age-standardized annual thyroidectomy rate in both sexes. We observed a large increase in the incidence of thyroid cancer, limited to papillary and early stage tumors, with a three- to four-fold parallel increase in thyroidectomy. The mortality slightly decreased. These findings suggest that a substantial and growing part of the detected thyroid cancers are overdiagnosed and overtreated. Targeted screening and diagnostic strategies are warranted to avoid overdetection and unnecessary treatment of thyroid cancers.


Learning of new sound categories shapes neural response patterns in human auditory cortex.
The formation of new sound categories is fundamental to everyday goal-directed behavior. Categorization requires the abstraction of discrete classes from continuous physical features as required by context and task. Electrophysiology in animals has shown that learning to categorize novel sounds alters their spatiotemporal neural representation at the level of early auditory cortex. However, functional magnetic resonance imaging (fMRI) studies so far did not yield insight into the effects of category learning on sound representations in human auditory cortex. This may be due to the use of overlearned speech-like categories and fMRI subtraction paradigms, leading to insufficient sensitivity to distinguish the responses to learning-induced, novel sound categories. Here, we used fMRI pattern analysis to investigate changes in human auditory cortical response patterns induced by category learning. We created complex novel sound categories and analyzed distributed activation patterns during passive listening to a sound continuum before and after category learning. We show that only after training, sound categories could be successfully decoded from early auditory areas and that learning-induced pattern changes were specific to the category-distinctive sound feature (i.e., pitch). Notably, the similarity between fMRI response patterns for the sound continuum mirrored the sigmoid shape of the behavioral category identification function. Our results indicate that perceptual representations of novel sound categories emerge from neural changes at early levels of the human auditory processing hierarchy.


ERP correlates of word production before and after stroke in an aphasic patient.
Changes in brain activity characterizing impaired speech production after brain damage have usually been investigated by comparing aphasic speakers with healthy subjects because prestroke data are normally not available. However, when interpreting the results of studies of stroke patients versus healthy controls, there is an inherent difficulty in disentangling the contribution of neuropathology from other sources of between-subject variability. In the present work, we had an unusual opportunity to study an aphasic patient with severe anomia who had incidentally performed a picture naming task in an ERP study as a control subject one year before suffering a left hemisphere stroke. The fortuitous recording of this patient's brain activity before his stroke allows direct comparison of his pre- and poststroke brain activity in the same language production task. The subject did not differ from other healthy subjects before his stroke, but presented major electrophysiological differences after stroke, both in comparison to himself before stroke and to the control group. ERP changes consistently appeared after stroke in a specific time window starting about 250 msec after picture onset, characterized by a single divergent but stable topographic configuration of the scalp electric field associated with a cortical generator abnormally limited to left temporal posterior perilesional areas. The patient's pattern of anomia revealed a severe lexical-phonological impairment and his ERP responses diverged from those of healthy controls in the time window that has previously been associated with lexical-phonological processes during picture naming. Given that his prestroke ERPs were indistinguishable from those of healthy controls, it seems highly likely that the change in his poststroke ERPs is due to changes in language production processes as a consequence of stroke. The patient's neurolinguistic deficits, combined with the ERPs results, provide unique evidence for the role of left temporal cortex in lexical-phonological processing from about 250 to 450 msec during word production.


Adoptive transfer of in vitro-targeted, activated T lymphocytes results in total tumor regression.
Adoptive immunotherapy of cancer uses the transfer of tumor-reactive immune cells. The success of this procedure is dependent upon the specificity of the transferred immune cells, their number, and their ability to reach their target cells. We genetically modified T lymphocytes and equipped them with the ability to specifically recognize tumor cells. Tumor cells overexpressing the ErbB-2 receptor served as a model. The target cell recognition specificity is conferred to T lymphocytes by transduction of a chimeric gene encoding the zeta-chain of the TCR and a single chain antibody (scFv(FRP5)) directed against the human ErbB-2 receptor. The chimeric scFv(FRP5)-zeta gene was introduced into primary mouse T lymphocytes via retroviral gene transfer. Naive T lymphocytes were activated and infected by cocultivation with a retrovirus-producing packaging cell line. The scFv(FRP5)-zeta fusion gene was expressed in >75% of the T cells. These T cells lysed ErbB-2-expressing target cells in vitro with high specificity. We tested the antitumor efficacy of scFv(FRP5)-zeta-expressing T cells in a syngeneic BALB/c model. The mice were treated with autologous, transduced T cells. The adoptively transferred scFv(FRP5)-zeta-expressing T cells caused total regression of ErbB-2-expressing tumors. The presence of the transduced T lymphocytes in the tumor tissue was monitored. No humoral response directed against the transduced T cells was observed. Abs directed against the ErbB-2 receptor were detected upon tumor lysis.


Development and Application of the Lincoln Adherence Instrument Record for Assessing Client Adherence to Advice in Dog Behavior Consultations and Success.
Adherence to the advice of medical practitioners is critical to successful treatment outcomes and has been much researched in human health, but is less well studied in the veterinary and clinical animal behavior fields. Given that the management of behavior problems often requires substantial change in established client behavior, it is likely that adherence is a substantive issue affecting success. However, little is known about the relationships between relevant factors, and there is no established way of assessing these. Therefore, the aim of this study was to develop an instrument for coding factors likely to impinge on pet owner adherence to behavior advice and validate its utility through the identification of the factors appearing to relate most closely to a successful treatment outcome in a sample population from our clinic. Potential factors affecting adherence were identified from human health and animal behavior studies, and a survey instrument developed with items matched to these factors. Forty-two dog owners who had attended the University of Lincoln Animal Behavior Clinic over a 2-year period provided data used in the analysis. The assessment of treatment outcome success by clients and clinicians was correlated, but clinicians tended to overestimate success by half a point on a 5-point scale. Eleven items relating to adherence were found to correlate with client ratings of treatment success in a univariate analysis, with three of these remaining in an ordinal logistic regression model. These three related to trust in the advice given by the clinician, concern over distress caused to the pet in the longer term and the perceived recommendation of treatment measures that had failed. By further examining the relationship between all of these factors in a hierarchical cluster analysis, we were able to postulate ways in which we might be able to improve client adherence and thus treatment success. This provides a model for the application of the instrument in any veterinary behavior practice wishing to use client feedback to rationalize areas of the consultation which might be improved.