Determination of anabolic agents in dietary supplements by liquid chromatography-high-resolution mass spectrometry.
A sensitive method for the identification and quantification of anabolic steroids and clenbuterol at trace levels in dietary supplements by liquid chromatography-high-resolution mass spectrometry (LC-HRMS) in atmospheric pressure ionisation (APCI) mode using a single-stage Orbitrap analyser operating at a resolution power of 100 000 full width at half maximum (FWHM) was developed and validated. A total of 1 g of dietary supplement was added with testosterone-d3 as internal standard, dissolved in methanol, evaporated to dryness, diluted in sodium hydroxide solution and extracted with a mixture of pentane/ethyl ether 9:1. The extract was directly injected into the LC-HRMS system. The method was fully validated. Limits of detection (LODs) obtained for anabolic androgenic steroids (AASs) varied from 1 to 25 ng g(-1) and the limit of quantitation (LOQ) was 50 ng g(-1) for all analytes. The calibration was linear for all compounds in the range from the LOQ to 2000 ng g(-1), with correlation coefficients always higher than 0.99. Accuracy (intended as %E) and repeatability (%CV) were always lower than 15%. Good values of matrix effect and recovery were achieved. The ease of the sample preparation together with a fast run time of only 16 min permitted rapid identification of the analytes. The method was applied to the analysis of 30 dietary supplements in order to check for the presence of anabolic agents not labelled as being present in these supplements. Many AASs were often detected in the same sample: indeed, androstenedione was detected in nine supplements, 5-androsten-3β-ol-17-one (DHEA) in 12, methandienone in three, stanozolol in one, testosterone in seven and testosterone esters in four of them. A retrospective analysis of suspected compounds not included at the beginning of the method development was also possible by means of the full acquisition spectra obtained with the HRMS technique.


Pro-Leu-glycinamide and its peptidomimetic, PAOPA, attenuate haloperidol induced vacuous chewing movements in rat: A model of human tardive dyskinesia.
In the present experimental paradigm, we examine the effect of L-prolyl-L-leucyl-glycinamide (PLG) co-administration with haloperidol on vacuous chewing movements (VCM) in rats-a model of tardive dyskinesia (TD) in humans. We examined the dose dependent induction of VCM through both injected and orally administered PLG (MIF-1). Our results show significant levels of VCM attenuation (P<0.05) in rats treated with 10mg/kg of PLG. Doses of 1 and 100mg/kg were ineffective. Reductions were present in both orally treated and injected rats. We also examined the therapeutic effect of a peptidomimetic of PLG-PAOPA. PAOPA was able to produce similar behavioral effects to PLG at a dose, which was 100-fold lower than the effective dose of PLG. These results suggest that PLG may play a role in D2 receptor expression and function, as well as providing a therapy for neuroleptic induced TD.


HookNet: Multi-resolution convolutional neural networks for semantic segmentation in histopathology whole-slide images.
We propose HookNet, a semantic segmentation model for histopathology whole-slide images, which combines context and details via multiple branches of encoder-decoder convolutional neural networks. Concentric patches at multiple resolutions with different fields of view, feed different branches of HookNet, and intermediate representations are combined via a hooking mechanism. We describe a framework to design and train HookNet for achieving high-resolution semantic segmentation and introduce constraints to guarantee pixel-wise alignment in feature maps during hooking. We show the advantages of using HookNet in two histopathology image segmentation tasks where tissue type prediction accuracy strongly depends on contextual information, namely (1) multi-class tissue segmentation in breast cancer and, (2) segmentation of tertiary lymphoid structures and germinal centers in lung cancer. We show the superiority of HookNet when compared with single-resolution U-Net models working at different resolutions as well as with a recently published multi-resolution model for histopathology image segmentation. We have made HookNet publicly available by releasing the source code1 as well as in the form of web-based applications2,3 based on the grand-challenge.org platform.


Expression of matrix metalloproteinases and tissue inhibitor of metalloproteinases in laryngeal and pharyngeal squamous cell carcinoma: A quantitative analysis.
Squamous cell carcinomas of the head and neck are known for their aggressive growth and propensity to metastasize. Invasion is facilitated by matrix metalloproteineases (MMPs). Tissue inhibitors of MMPs (TIMPs) negatively regulate MMP activity. MMP and TIMP expression in head and neck squamous cell carcinomas was determined by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). qRT-PCR allows measurement of several mRNAs from as little as 4 microg of total cellular RNA. We measured MMP-1, MMP-2, MMP-9, and TIMP-1 expression in 8 specimens of primary tumors and adjacent normal tissue. MMP-1 was overexpressed in 6 of 8 tumors, and MMP-9 was overexpressed in 4 of 7 tumors. MMP-2 was expressed in 3 of 8 tumors and 3 of 8 normal samples. TIMP-1 was expressed in all specimens. This work demonstrates that qRT-PCR can be used to examine expression of specific mRNAs in clinical specimens. Therefore this method provides another tool for the molecular analysis of tumors.


Elevated Homocysteine Levels and Hypertension Relate to Cognitive Impairment via Increased White Matter Hyperintensity Volume.
Recent studies have identified a relationship between elevated homocysteine levels and hypertension (HTN) with Alzheimer's disease (AD), but its pathogenesis remains unclear. To evaluate elevated homocysteine levels and HTN as risk factors for cognitive impairment (CI) and determine their relationship with white matter hyperintensity (WMH) volume. A total of 521 subjects were selected from the Alzheimer's Disease Neuroimaging Initiative (ADNI) database and divided into two groups according to the diagnostic criteria of the ADNI database. The CI group included 370 subjects, consisting of 122 with AD and 248 with mild CI, while the cognitively normal (CN) group contained 151 subjects. The history of HTN, homocysteine levels, WMH volume and Mini-Mental State Examination (MMSE) scores were analyzed. The study found that patients with CI had higher homocysteine levels than those with CN. Additionally, WMH volume was significantly correlated with homocysteine levels in CI patients, and MMSE scores decreased as WMH volume increased. Further analysis revealed that CI patients with HTN had significantly higher homocysteine levels than those without HTN. Furthermore, the correlation between WMH volume and homocysteine levels was significant only in CI patients with HTN and not in those without HTN. In CN patients, there was no correlation between WMH volume and homocysteine levels in either the HTN or non-HTN groups, and no difference was observed in homocysteine levels. It is indicated that elevated homocysteine levels in conjunction with HTN are associated with the increased volume of WMHs and CI.


Effect of selective monoamine oxidase inhibition by clorgyline and deprenyl on the norepinephrine receptor-coupled adenylate cyclase system in rat cortex.
The consequences of selective monoamine oxidase (MAO) inhibition on the norepinephrine(NE)-sensitive adenylate cyclase system were determined in slices of rat cerebral cortex. The chronic administration of clorgyline, which selectively inhibited the activity of MAO-A, caused a significant decrease in the responsiveness of the noradrenergic cyclic AMP-generating system. The noradrenergic subsensitivity was accompanied by a significant decrease in the density of beta-adrenoceptors, as measured by 3H-dihydroalprenolol (DHA) binding, without altering the Kd value. However, selective inhibition of MAO-B by deprenyl did not alter the sensitivity of the cyclic AMP-generating system to NE or the specific DHA binding. The basal levels of cyclic AMP in the cortex were unaltered by the drugs. Since inhibition of MAO-A, but not MAO-B, increases the availability of NE, the results support the hypothesis that a persistent NE-receptor interaction is one of the prerequisites for the in vivo densitization of the NE-sensitive adenylate cyclase and the concomitant down-regulation of the number of beta-adrenoceptors in brain.


Synthesis and bio-functionalization of magnetic nanoparticles for medical diagnosis and treatment.
The synthesis of multifunctional magnetic nanoparticles (NPs) is a highly active area of current research located at the interface between materials science, biotechnology and medicine. By virtue of their unique physical properties magnetic nanoparticles are emerging as a new class of diagnostic probes for multimodal tracking and as contrast agents for MRI. Furthermore, they show great potential as carriers for targeted drug and gene delivery, since reactive agents, such as drug molecules or large biomolecules (including genes and antibodies), can easily be attached to their surface. On the other hand, the fate of the nanoparticles inside the body is mainly determined by the interactions with its local environment. These interactions strongly depend upon the size of the magnetic NPs but also on the individual surface characteristics, like charge, morphology and surface chemistry. This review not only summarizes the most common synthetic approaches for the generation of magnetic NPs, it also focuses on different surface modification strategies that are used today to enhance the biocompatibility of these NPs. Finally, key considerations for the application of magnetic NPs in biomedicine, as well as various examples for the utilization in multimodal imaging and targeted gene delivery are presented.


Vitamin D Status in Hospitalized Patients with SARS-CoV-2 Infection.
The role of vitamin D status in COVID-19 patients is a matter of debate. To assess serum 25-hydroxyvitamin D (25OHD) levels in hospitalized patients with COVID-19 and to analyze the possible influence of vitamin D status on disease severity. Retrospective case-control study of 216 COVID-19 patients and 197 population-based controls. Serum 25OHD levels were measured in both groups. The association of serum 25OHD levels with COVID-19 severity (admission to the intensive care unit, requirements for mechanical ventilation, or mortality) was also evaluated. Of the 216 patients, 19 were on vitamin D supplements and were analyzed separately. In COVID-19 patients, mean ± standard deviation 25OHD levels were 13.8 ± 7.2 ng/mL, compared with 20.9 ± 7.4 ng/mL in controls (P < .0001). 25OHD values were lower in men than in women. Vitamin D deficiency was found in 82.2% of COVID-19 cases and 47.2% of population-based controls (P < .0001). 25OHD inversely correlates with serum ferritin (P = .013) and D-dimer levels (P = .027). Vitamin D-deficient COVID-19 patients had a greater prevalence of hypertension and cardiovascular diseases, raised serum ferritin and troponin levels, as well as a longer length of hospital stay than those with serum 25OHD levels ≥20 ng/mL. No causal relationship was found between vitamin D deficiency and COVID-19 severity as a combined endpoint or as its separate components. 25OHD levels are lower in hospitalized COVID-19 patients than in population-based controls and these patients had a higher prevalence of deficiency. We did not find any relationship between vitamin D concentrations or vitamin deficiency and the severity of the disease.


Exploring Force Production Reliability across Different Levels of Clinical Experience during a Simulated One-handed Instrument-Assisted Soft Tissue Mobilization Treatment: A Pilot Study.
Instrument-assisted soft tissue mobilization (IASTM) is a commonly utilized intervention for musculoskeletal pain and dysfunction. However, little is known regarding the reliability of forces applied by clinicians of different experience levels during an IASTM intervention. The purpose of this pilot study was to assess intra-clinician reliability of IASTM force (i.e., mean normal force) during a simulated, one-handed stroke IASTM intervention across different levels of IASTM clinical experience. Descriptive laboratory study. The researchers conducted a repeated measures trial in a laboratory setting with a convenience sample of ten participants who had previously completed professional IASTM training. Participants performed 15 one-handed sweeping strokes with an IASTM instrument on a skin simulant attached to a force plate for a standardized hypothetical treatment scenario. The participants performed the treatment on two separate days, 24-48 hours apart. The researchers examined the intra-rater reliability for average (mean) normal forces using Bland-Altman (BA) plots and Coefficient of Variation (CV) values. The BA plot results indicated all participants (professional athletic training students = 4, athletic trainers = 6; males = 5, females = 5; age = 32.60 ± 8.71 y; IASTM experience = 3.78 ± 4.10 y), except participant D (1.9N, 190g), were consistently reliable within 1N (100g) or less of force for mean differences and within the maximum limits of agreement around 3.7N (370g). Most participants' CV scores ranged between 8 to 20% supporting reliable force application within each treatment session. The data indicated that IASTM trained clinicians could produce consistent forces within and across treatment sessions irrespective of clinical experience. 3.


Protein synthesis- and fucosylation-dependent mechanisms in corticosterone facilitation of long-term memory in the chick.
Long-term memory formation for a passive avoidance task in day-old chicks, which requires a late phase of glycoprotein fucosylation (5-8 hr posttraining), is dependent on a corticosterone action in the brain. In addition, corticosterone enhances late-phase fucosylation. In this study, the authors explored (a) to what extent the memory-facilitating action of corticosterone is dependent on protein fucosylation and (b) whether a protein synthesis mechanism might be involved in the steroid effects on memory and late-phase fucosylation. A combination of psychopharmacological and biochemical experiments, including the fucosylation inhibitor 2-deoxygalactose (2-DG), the protein synthesis inhibitor anisomycin (ANI), and radiolabeled fucose, indicated that the late phase of glycoprotein synthesis involved in the memory-facilitating effect of corticosterone occurs on newly synthesized proteins.


Vitamin E Enhances Cancer Immunotherapy by Reinvigorating Dendritic Cells via Targeting Checkpoint SHP1.
Despite the popular use of dietary supplements during conventional cancer treatments, their impacts on the efficacies of prevalent immunotherapies, including immune-checkpoint therapy (ICT), are unknown. Surprisingly, our analyses of electronic health records revealed that ICT-treated patients with cancer who took vitamin E (VitE) had significantly improved survival. In mouse models, VitE increased ICT antitumor efficacy, which depended on dendritic cells (DC). VitE entered DCs via the SCARB1 receptor and restored tumor-associated DC functionality by directly binding to and inhibiting protein tyrosine phosphatase SHP1, a DC-intrinsic checkpoint. SHP1 inhibition, genetically or by VitE treatment, enhanced tumor antigen cross-presentation by DCs and DC-derived extracellular vesicles (DC-EV), triggering systemic antigen-specific T-cell antitumor immunity. Combining VitE with DC-recruiting cancer vaccines or immunogenic chemotherapies greatly boosted ICT efficacy in animals. Therefore, combining VitE supplement or SHP1-inhibited DCs/DC-EVs with DC-enrichment therapies could substantially augment T-cell antitumor immunity and enhance the efficacy of cancer immunotherapies. The impacts of nutritional supplements on responses to immunotherapies remain unexplored. Our study revealed that dietary vitamin E binds to and inhibits DC checkpoint SHP1 to increase antigen presentation, prime antitumor T-cell immunity, and enhance immunotherapy efficacy. VitE-treated or SHP1-silenced DCs/DC-EVs could be developed as potent immunotherapies. This article is highlighted in the In This Issue feature, p. 1599.


Do the pyramids show continental drift?
The mystery of the orientation of the Great Pyramids of Giza has remained unexplained for many decades. The general alignment is 4 minutes west of north. It is argued that this is not a builders' error but is caused by movement over the centuries. Modern theories of continental drift do not predict quite such large movements, but other causes of polar wandering give even smaller shifts. Thus, continental drift is the most likely explanation, although somewhat implausible, especially as relevant measurements have been made over a 50-year period, whereas geophysical measurements of sea-floor spreading relate to million-year time scales.


"What Is the Benefit?": Perceptions and Preferences for Long-Acting Injectable Antiretroviral Therapy Among People Living With HIV.
Long-acting injectable antiretroviral therapy (LA-ART) expands treatment options for people living with HIV (PLWH). This qualitative study characterizes LA-ART awareness, perceptions, and preferences among PLWH engaged in HIV care. From 2019 through 2021, we conducted semistructured in-depth interviews with 71 PLWH sampled from three clinics in three U.S. settings (North Carolina, Washington, DC, Massachusetts). Transcripts were analyzed using narrative and thematic techniques. Participant mean age was 46 years (range 24-72); most were cisgender men (55%) and virally suppressed (73%). Most participants had not heard of LA-ART and reacted with a mix of excitement and cautiousness. Potential LA-ART benefits included easier adherence, privacy, and effectiveness; concerns included effectiveness, side effects, costs, and increased clinic visits. Participants appreciated that LA-ART could support achieving and sustaining viral suppression. To inform their decision, participants wanted more information and convenient access and administration. Findings indicated that a shared decision-making approach and economic and logistical support for PLWH could facilitate LA-ART uptake.


The limits of aerial twisting techniques in the aerials event of freestyle skiing.
In the aerials event of freestyle skiing, athletes perform three somersaults with up to five twists. This study investigated the twisting limits of such movements using a computer simulation model of aerial movement. The abilities of various asymmetrical arm and hip techniques to produce twist during flight were investigated using 10 simulations to maximise twist and allow reorientation prior to landing. It was found that 4-6 twists could be produced during three somersaults. The main limiting factor was the increased whole body frontal moment of inertia due to the equipment which restricted the amount of tilt resulting from an asymmetrical arm movement. It was concluded that reductions in equipment mass might make such movements easier to achieve but would be unlikely to allow advances beyond the limits found.


Pathophysiological considerations in left ventricular hypertrophy.
Considerable evidence is available to relate development of left ventricular hypertrophy to the maintenance and progression of hypertensive vascular disease and its associated hemodynamic changes that induce an increased left ventricular afterload. However, when critical investigators question in depth the variety of other nonhemodynamic factors that may also be associated with the development (or regression) of left ventricular hypertrophy, they are faced with three disquieting conclusions. First, knowledge of those mechanisms that account for the development and maintenance (or, for that matter, even regression) of left ventricular hypertrophy in hypertension is incomplete. Second, little is known as to how the physical hemodynamic and mechanical factors associated with the pathogenesis of hypertension are translated into the biochemical events associated with the development of cardiac muscle hypertrophy. Finally, to understand these important factors more clearly, the investigator (clinical or experimental) must design studies more precisely to control for the additional complicating, nonhemodynamic factors.


[The role of double-ballon enteroscopy in diagnosis and treatment of polypoid syndromes of gastrointestinal tract].
It was done 192 double-ballon enteroscopies in 141 patients at the period from March 2009 to March 2012. Epithelial benign tumors were detected in 10 (7.1%) patients with congenital polypoid syndromes. Endoscopic excision of polyps was done in 6 patients including 1 case with laparoscopic-assisted technique. Urgent open surgeries were performed before this in 4 patients with Peutz-Jeghers' syndrome and invaginated small bowel obstruction. The authors consider that double-ballon enteroscopy is highly informative diagnostic and treatment method in case of small bowel diseases. It allows to diagnose and to treat epithelial tumors on the early stage. Also it could help to avoid complications development and disability of patients.


Myocardial adaptations to endurance exercise in aged rats.
Isolated perfused working hearts of 25-mo-old male Sprague-Dawley rats trained to run for 16 wk were compared with hearts from 9- and 25-mo-old sedentary animals. Under low work load conditions, systolic and diastolic aortic pressures, aortic flow, and oxygen consumption of the three groups were similar. Under high work load, systolic pressure of trained old and 9-mo groups were higher than the 25-mo sedentary values, but diastolic pressures were similar. At a systolic pressure of 150 mmHg, coronary flow of the old trained heart was higher than that of the age-matched controls, although not equal to the 9-mo sedentary group. The oxygen consumption of the intact hearts under the latter conditions follows the same quantitative trend. Left ventricular cytochrome c concentrations and rates of oxidation of glutamate-malate, palmitoylcarnitine, and succinate were increased in the older rats by training but not to the level of the 9-mo old. These data indicate that appropriate exercise in aged animals improves myocardial function and aerobic energy metabolism.


Novel therapeutic for multiple sclerosis protects white matter function in EAE mouse model.
Multiple sclerosis (MS) is a chronic demyelinating disease with prominent axon dysfunction. Our previous studies in an MS mouse model, experimental autoimmune encephalomyelitis (EAE), demonstrated that major histocompatibility complex Class II constructs can reverse clinical signs of EAE. These constructs block binding and downstream signaling of macrophage migration inhibitory factors (MIF-1/2) through CD74, thereby inhibiting phosphorylation of extracellular signal-regulated kinase (ERK) activation and tissue inflammation and promoting remyelination. To directly assess the effects of a novel third generation construct, DRhQ, on axon integrity in EAE, we compared axon conduction properties using electrophysiology on corpus callosum slices and optic nerves. By using two distinct white matter (WM) tracts, we aimed to assess the impact of the EAE and the benefit of DRhQ on myelinated and unmyelinated axons as well as to test the clinical value of DRhQ on demyelinating lesions in CC and optic myelitis. Our study found that EAE altered axon excitability, delayed axon conduction and slowed spatiotemporal summation correlated with diffuse astrocyte and microglia activation. Because MS predisposes patients to stroke, we also investigated and showed that vulnerability to WM ischemia is increased in the EAE MS mouse model. Treatment with DRhQ after the onset of EAE drastically inhibited microglial and astrocyte activation, improved functional integrity of the myelinated axons and enhanced recovery after ischemia. These results demonstrate that DRhQ administered after the onset of EAE promotes WM integrity and function, and reduces subsequent vulnerability to ischemic injury, suggesting important therapeutic potential for treatment of progressive MS.


Altered lung structure and function in mid-childhood survivors of very preterm birth.
Survivors of preterm birth are at risk of chronic and lifelong pulmonary disease. Follow-up data describing lung structure and function are scarce in children born preterm during the surfactant era. To obtain comprehensive data on lung structure and function in mid-childhood from survivors of preterm birth. We aimed to explore relationships between lung structure, lung function and respiratory morbidity as well as early life contributors to poorer childhood respiratory outcomes. Lung function was tested at 9-11 years in children born at term (controls) and at ≤32 weeks gestation. Tests included spirometry, oscillatory mechanics, multiple breath nitrogen washout and diffusing capacity of the lung for carbon monoxide. Preterm children had CT of the chest and completed a respiratory symptoms questionnaire. 58 controls and 163 preterm children (99 with bronchopulmonary dysplasia) participated. Preterm children exhibited pulmonary obstruction and hyperinflation as well as abnormal peripheral lung mechanics compared with term controls. FEV1 was improved by 0.10 z-scores for every additional week of gestation (95% CI 0.028 to 0.182; p=0.008) and by 0.34 z-scores per z-score increase in birth weight (0.124 to 0.548; p=0.002). Structural lung changes were present in 92% of preterm children, with total CT score decreased by 0.64 (-0.99 to -0.29; p<0.001) for each additional week of gestation. Obstruction was associated with increased subpleural opacities, bronchial wall thickening and hypoattenuated lung areas on inspiratory chest CT scans (p<0.05). Abnormal lung structure in mid-childhood resulting from preterm birth in the contemporary era has important functional consequences.


Negative Ions in Space.
Until a decade ago, the only anion observed to play a prominent role in astrophysics was H-. The bound-free transitions in H- dominate the visible opacity in stars with photospheric temperatures less than 7000 K, including the Sun. The H- anion is also believed to have been critical to the formation of molecular hydrogen in the very early evolution of the Universe. Once H2 formed, about 500 000 years after the Big Bang, the expanding gas was able to lose internal gravitational energy and collapse to form stellar objects and "protogalaxies", allowing the creation of heavier elements such as C, N, and O through nucleosynthesis. Although astronomers had considered some processes through which anions might form in interstellar clouds and circumstellar envelopes, including the important role that polycyclic aromatic hydrocarbons might play in this, it was the detection in 2006 of rotational line emission from C6H- that galvanized a systematic study of the abundance, distribution, and chemistry of anions in the interstellar medium. In 2007, the Cassini mission reported the unexpected detection of anions with mass-to-charge ratios of up to ∼10 000 in the upper atmosphere of Titan; this observation likewise instigated the study of fundamental chemical processes involving negative ions among planetary scientists. In this article, we review the observations of anions in interstellar clouds, circumstellar envelopes, Titan, and cometary comae. We then discuss a number of processes by which anions can be created and destroyed in these environments. The derivation of accurate rate coefficients for these processes is an essential input for the chemical kinetic modeling that is necessary to fully extract physics from the observational data. We discuss such models, along with their successes and failings, and finish with an outlook on the future.


SARS-CoV-2 Testing in Florida, Illinois, and Maryland: Access and Barriers.
To characterize the SARS-CoV-2 testing cascade and associated barriers in three US states. We recruited participants from Florida, Illinois, and Maryland (~1000/state) for an online survey September 16 - October 15, 2020. The survey covered demographics, COVID-19 symptoms, and experiences around SARS-CoV-2 PCR testing in the prior 2 weeks. Logistic regression was used to analyze associations with outcomes of interest. Overall, 316 (10%) of 3,058 respondents wanted/needed a test in the two weeks prior to the survey. Of these, 166 (53%) were able to get tested and 156 (94%) received results; 53% waited ≥ 8 days to get results from when they wanted/needed a test. There were no significant differences by state. Among those wanting/needing a test, getting tested was significantly less common among men (aOR: 0.46) and those reporting black race (aOR: 0.53) and more common in those reporting recent travel (aOR: 3.35). There is an urgent need for a national communication strategy on who should get tested and where one can get tested. Additionally, measures need to be taken to improve access and reduce turn-around-time.


Mutual Shaping in Swarm Robotics: User Studies in Fire and Rescue, Storage Organization, and Bridge Inspection.
Many real-world applications have been suggested in the swarm robotics literature. However, there is a general lack of understanding of what needs to be done for robot swarms to be useful and trusted by users in reality. This paper aims to investigate user perception of robot swarms in the workplace, and inform design principles for the deployment of future swarms in real-world applications. Three qualitative studies with a total of 37 participants were done across three sectors: fire and rescue, storage organization, and bridge inspection. Each study examined the users' perceptions using focus groups and interviews. In this paper, we describe our findings regarding: the current processes and tools used in these professions and their main challenges; attitudes toward robot swarms assisting them; and the requirements that would encourage them to use robot swarms. We found that there was a generally positive reaction to robot swarms for information gathering and automation of simple processes. Furthermore, a human in the loop is preferred when it comes to decision making. Recommendations to increase trust and acceptance are related to transparency, accountability, safety, reliability, ease of maintenance, and ease of use. Finally, we found that mutual shaping, a methodology to create a bidirectional relationship between users and technology developers to incorporate societal choices in all stages of research and development, is a valid approach to increase knowledge and acceptance of swarm robotics. This paper contributes to the creation of such a culture of mutual shaping between researchers and users, toward increasing the chances of a successful deployment of robot swarms in the physical realm.


Triggering of the CD44 antigen on T lymphocytes promotes T cell adhesion through the LFA-1 pathway.
The CD44 molecule, a molecule which has been previously known as Hermes, Pgp-1, extracellular matrix receptor III, and In(Lu)-related p80, is currently thought to be involved in several steps of normal immune cell function, including lymphocyte adhesion to high endothelial venules and to the extracellular matrix and T cell activation. We now demonstrate that triggering of CD44 on T lymphocytes by anti-CD44 mAb promotes cell adhesion. The induced homotypic adhesion is mediated by lymphocyte function-associated antigen-1 (LFA-1), because it was inhibited by anti-LFA-1 antibodies and not by anti-LFA-3 antibodies. This notion is supported by the temperature and Mg2+ dependence which is characteristic of LFA-1-mediated adhesion. Moreover, the sensitivity of CD44-induced adhesion to AMG and H7, which both prevent the activation of protein kinase C, and to cytochalasin B, which inhibits microfilament formation, suggests that the activation of the LFA-1 pathway via CD44 involves protein kinase C activation and requires an intact cytoskeleton.


Development of liquid culture media mimicking the conditions of sinuses and lungs in cystic fibrosis and health.
The respiratory tract is a compartmentalised and heterogenous environment. The nasopharynx and sinuses of the upper airways have distinct properties from the lungs and these differences may shape bacterial adaptation and evolution. Upper airway niches act as early colonisation sites for respiratory bacterial pathogens, including those, such as Pseudomonas aeruginosa, that can go on to establish chronic infection of the lungs in people with cystic fibrosis (CF). Despite the importance of upper airway environments in facilitating early adaptation to host environments, currently available in vitro models for study of respiratory infection in CF focus exclusively on the lungs. Furthermore, animal models, widely used to bridge the gap between in vitro systems and the clinical scenario, do not allow the upper and lower airways to be studied in isolation. We have developed a suite of culture media reproducing key features of the upper and lower airways, for the study of bacterial adaptation and evolution in different respiratory environments. For both upper and lower airway-mimicking media, we have developed formulations that reflect airway conditions in health and those that reflect the altered environment of the CF respiratory tract. Here, we describe the development and validation of these media and their use for study of genetic and phenotypic adaptations in P. aeruginosa during growth under upper or lower airway conditions in health and in CF.


Brain connectivity in neurodegenerative diseases--from phenotype to proteinopathy.
Functional and structural connectivity measures, as assessed by means of functional and diffusion MRI, are emerging as potential intermediate biomarkers for Alzheimer disease (AD) and other disorders. This Review aims to summarize current evidence that connectivity biomarkers are associated with upstream and downstream disease processes (molecular pathology and clinical symptoms, respectively) in the major neurodegenerative diseases. The vast majority of studies have addressed functional and structural connectivity correlates of clinical phenotypes, confirming the predictable correlation with topography and disease severity in AD and frontotemporal dementia. In neurodegenerative diseases with motor symptoms, structural--but, to date, not functional--connectivity has been consistently found to be associated with clinical phenotype and disease severity. In the latest studies, the focus has moved towards the investigation of connectivity correlates of molecular pathology. Studies in cognitively healthy individuals with brain amyloidosis or genetic risk factors for AD have shown functional connectivity abnormalities in preclinical disease stages that are reminiscent of abnormalities observed in symptomatic AD. This shift in approach is promising, and may aid identification of early disease markers, establish a paradigm for other neurodegenerative disorders, shed light on the molecular neurobiology of connectivity disruption and, ultimately, clarify the pathophysiology of neurodegenerative diseases.


Reward Modulates Visual Responses in the Superficial Superior Colliculus of Mice.
The processing of sensory input is constantly adapting to behavioral demands and internal states. The drive to obtain reward, e.g., searching for water when thirsty, is a strong behavioral demand and associating the reward with its source, a certain environment or action, is paramount for survival. Here, we show that water reward increases subsequent visual activity in the superficial layers of the superior colliculus (SC), which receive direct input from the retina and belong to the earliest stages of visual processing. We trained mice of either sex to perform a visual decision task and recorded the activity of neurons in the SC using two-photon calcium imaging and high-density electrophysiological recordings. Responses to visual stimuli in around 20% of visually responsive neurons in the superficial SC were affected by reward delivered in the previous trial. Reward mostly increased visual responses independent from modulations due to pupil size changes. The modulation of visual responses by reward could not be explained by movements like licking. It was specific to responses to the following visual stimulus, independent of slow fluctuations in neural activity and independent of how often the stimulus was previously rewarded. Electrophysiological recordings confirmed these results and revealed that reward affected the early phase of the visual response around 80 ms after stimulus onset. Modulation of visual responses by reward, but not pupil size, significantly improved the performance of a population decoder to detect visual stimuli, indicating the relevance of reward modulation for the visual performance of the animal.SIGNIFICANCE STATEMENT To learn which actions lead to food, water, or safety, it is necessary to integrate the receiving of reward with sensory stimuli related to the reward. Cortical stages of sensory processing have been shown to represent stimulus-reward associations. Here, we show, however, that reward influences neurons at a much earlier stage of sensory processing, the superior colliculus (SC), receiving direct input from the retina. Visual responses were increased shortly after the animal received the water reward, which led to an improved stimulus signal in the population of these visual neurons. Reward modulation of early visual responses may thus improve perception of visual environments predictive of reward.


Effects of sudden changes in external sodium concentration on twitch tension in isolated muscle fibers.
When [Na] was suddenly introduced to single muscle fibers (Xenopus or frog), which had been pretreated with Na-free solution (Tris-substituted), the time-course of twitch recovery was very variable, half-time ranging from less than 1 S to 5 S. The [Na] vs. twitch height relationship was also variable. In small Xenopus fibers, decreases of [Na] to 50% increased the twitch, while in large Xenopus fibers twitch height remained constant or decreased as [Na] was decreased to 50%. The apparent diffusion constant (D') of Na+ or K+, calculated from the time-course of twitch recovery and the [Na] vs. twitch relation, and from the time-course of the slow repolarization upon sudden reduction of [K] was about 1-1.5 X 10(-6) cm2/S. This is one order of magnitude smaller than the diffusion constants in an aqueous solution. Even if the tortuosity factor of the T system is taken into account, there remains a substantial discrepancy. Although our value of D' is subject to various errors, if we accept the value, the twitch recovery is predicted to be either very quick or slow depending upon the variation of [Na]-twitch relation and fiber size. Thus, both quick and slow twitch recoveries can be explained by the diffusion time of Na+ in the T system, and therefore the results are consistent with the idea that the T system is excitable.


Characterization of changes in gene expression and biochemical pathways at low levels of benzene exposure.
Benzene, a ubiquitous environmental pollutant, causes acute myeloid leukemia (AML). Recently, through transcriptome profiling of peripheral blood mononuclear cells (PBMC), we reported dose-dependent effects of benzene exposure on gene expression and biochemical pathways in 83 workers exposed across four airborne concentration ranges (from <1 ppm to >10 ppm) compared with 42 subjects with non-workplace ambient exposure levels. Here, we further characterize these dose-dependent effects with continuous benzene exposure in all 125 study subjects. We estimated air benzene exposure levels in the 42 environmentally-exposed subjects from their unmetabolized urinary benzene levels. We used a novel non-parametric, data-adaptive model selection method to estimate the change with dose in the expression of each gene. We describe non-parametric approaches to model pathway responses and used these to estimate the dose responses of the AML pathway and 4 other pathways of interest. The response patterns of majority of genes as captured by mean estimates of the first and second principal components of the dose-response for the five pathways and the profiles of 6 AML pathway response-representative genes (identified by clustering) exhibited similar apparent supra-linear responses. Responses at or below 0.1 ppm benzene were observed for altered expression of AML pathway genes and CYP2E1. Together, these data show that benzene alters disease-relevant pathways and genes in a dose-dependent manner, with effects apparent at doses as low as 100 ppb in air. Studies with extensive exposure assessment of subjects exposed in the low-dose range between 10 ppb and 1 ppm are needed to confirm these findings.


PepsinA as a Marker of Laryngopharyngeal Reflux Detected in Chronic Rhinosinusitis Patients.
Objectives We aimed to confirm the presence of pepsinA in the nasal secretions and tissues of chronic rhinosinusitis (CRS) patients and reveal the relationship between CRS and laryngopharyngeal reflux (LPR). Study Design Cross-sectional study. Setting The study was conducted at the Department of Oto-Rhino-Laryngology, West China Hospital, Sichuan University. Subjects and Methods A total of 32 CRS patients with or without nasal polyps (CRSwNP and CRSsNP, respectively) and 10 normal controls were enrolled in our study. We investigated the expression of pepsinA in the nasal tissues, secretions, and blood plasma from the subjects by immunohistochemical staining, Western blot, or ELISA. Additionally, the expressions of MUC4, MUC5AC, MUC5B, MUC8, and pepsinogenA in nasal tissue were evaluated by quantitative real-time polymerase chain reaction. Results Immunohistochemistry and Western blot revealed that the pepsinA expression levels in the turbinate mucosa in CRSwNP/CRSsNP patients, which were largely restricted to the epithelial layer or glandular mucous cells in nasal tissues, were significantly higher than those in controls and in the polyp tissues of CRSwNP patients ( P < .05). In addition, the concentration of pepsinA in nasal secretions was significantly increased in the CRSwNP (147.85 ± 53.69 ng/mL, P < .001) and CRSsNP (134.12 ± 36.23 ng/mL, P < .001) groups as compared with the controls (68.69 ± 19.28 ng/mL). Although MUC5AC, MUC5B, and MUC8 expression differed among the groups, no correlation between pepsinA and mucin genes was found. Conclusion The results of this study provided evidence of an association between LPR and CRS, although no correlation was found to exist between LPR and mucin genes in CRS patients.


Azo dyes degradation and mutagenicity evaluation with a combination of microbiological and oxidative discoloration treatments.
This work evaluated the degradation of the Acid Blue 161 and Procion Red MX-5B dyes in a binary solution by the filamentous fungus Aspergillus terreus and the yeast Saccharomyces cerevisiae in systems with and without electrochemical oxidation as the pretreatment process. UV-Vis spectrophotometry, high-performance liquid chromatography with (HPLC), Fourier transform infrared (FT-IR) spectroscopy and Salmonella/microsome assay (Ames test) were applied towards the degradation analysis of the dyes. Adsorption tests with white clay immobilized on alginate were also conducted after the discoloration treatments to remove intermediate metabolites formed during the degradation of the dye molecules. The discoloration treatments led to the complete color removal of the solutions in all the systems tested. The clay demonstrated affinity for the metabolites formed after discoloration treatments, the removal rates were variable, but the all systems has proved efficient. The Salmonella/microsome assay (Ames test) with strains TA98 and TA100 in the absence and presence of exogenous metabolism (S9 microsomal system, Moltox) revealed that the initial molecules and by-products of the metabolism of the dyes were direct mutagens. The electrochemical/A. terreus/clay system was able to discolor the solutions and transform the direct mutagens into non-mutagenic compounds in addition to reducing the mutagenic potency of the pro-mutagens to the Salmonella strain TA100/S9, which demonstrates the high efficiency of this system with regard to discoloring and degrading azo dye molecules and their by-products. Therefore, this study showed that although not having standard treatment system for this type of pollutant, the combination of treatments can be considered promising. The use of electrochemical oxidation along with microbiological treatment may lead to the degradation and mineralization of these compounds, reducing or eliminating the environmental impact caused by the improper disposal of these dyes in aquatic environments.


[The tuberculous meningoencephalitis in patients with later stages of HIV infection].
Based on the examination and treatment of 120 patients with tuberculous meningoencephalitis (TM) in the later stages of HIV infection, the differences between these patients and patients without HIV infection were found. HIV-infected patients with TM had a more acute disease onset, more clinical symptoms of encephalitis confirmed by magnetic resonance imaging of the brain, more frequent presence of mycobacterium tuberculosis in the cerebrospinal fluid and higher level of resistance to tuberculosis drugs. The morphologic study demonstrated the domination of necrotic and exudative reactions over productive inflammation with destructive abscess-type lesions in the brain matter. The mortality was more than 2 times higher than that in the control group.


Clients' and therapists' views of the therapeutic alliance: similarities, differences and relationship to therapy outcome.
To better understand how clients' and therapists' views of the therapeutic alliance differ and overlap, this study investigated, first, the components of the alliance that are relevant to the therapy participants; second, their relationship to post-therapy outcome; and third, the relationships between participants' alliance constructs. To identify participants' views, exploratory factor analyses were performed on clients' (n = 176) and therapists' (n = 133 observations) ratings of the Working Alliance Inventory (short form), the Helping Alliance Questionnaire and the California Psychotherapy Alliance Scales and conducted both on each measure separately and on the three measures combined. The results of the separate analyses indicated in general poor correspondence between the participant-derived components and each measure's a priori constructs. Results of the joint analyses suggested that clients view the alliance in terms of six basic components (Collaborative Work Relationship, Productive Work, Active Commitment, Bond, Non-disagreement on Goals/Tasks and Confident Progress), five of which were found to predict client-rated and/or therapist-rated post-therapy outcome. Results for therapists suggested four basic components (Collaborative Work Relationship, Therapist Confidence & Dedication, Client Commitment & Confidence, Client Working Ability), of which three predicted post-therapy outcome. Findings of significant, but modest to low moderate, correlations between several client and therapist joint factors suggested that despite similarities, the therapy partners' views of the alliance differ in important ways. Compared with therapists, clients appear to place greater emphasis on helpfulness, joint participation in the work of therapy and negative signs of the alliance. Implications of these findings are discussed. Therapists should not assume that their views of the therapeutic relationship and therapeutic work are shared by their clients and are encouraged to seek the client's feedback. Therapists may benefit from conveying that the client's perspective on problems and relevant work is valued and that they are working with the client as a team. Therapists may need to explicitly address how the therapeutic work is helpful and conducive to desired changes.


Resistance training with creatine monohydrate improves upper-body strength in patients with Parkinson disease: a randomized trial.
Persons with Parkinson disease (PD) exhibit decreased muscular fitness including decreased muscle mass, muscle strength, bioenergetic capabilities and increased fatigability. This purpose of this investigation was to evaluate the therapeutic effects of resistance training with and without creatine supplementation in patients with mild to moderate PD. Twenty patients with idiopathic PD were randomized to receive creatine monohydrate supplementation plus resistance training (CRE) or placebo (lactose monohydrate) plus resistance training (PLA), using a double-blind procedure. Creatine and placebo supplementation consisted of 20 g/d for the first 5 days and 5 g/d thereafter. Both groups participated in progressive resistance training (24 sessions, 2 times per week, 1 set of 8-12 repetitions, 9 exercises). Participants performed 1-repetition maximum (1-RM) for chest press, leg extension, and biceps curl. Muscular endurance was evaluated for chest press and leg extension as the number of repetitions to failure using 60% of baseline 1-RM. Functional performance was evaluated as the time to perform 3 consecutive chair rises. Statistical analyses (ANOVA) revealed significant Group x Time interactions for chest press strength and biceps curl strength, and post hoc testing revealed that the improvement was significantly greater for CRE. Chair rise performance significantly improved only for CRE (12%, P=.03). Both PLA and CRE significantly improved 1-RM for leg extension (PLA: 16%; CRE: 18%). Muscular endurance improved significantly for both groups. These findings demonstrate that creatine supplementation can enhance the benefits of resistance training in patients with PD.


Efficacy and Safety of 3 Nasal Packing Materials Used After Functional Endoscopic Sinus Surgery for Chronic Rhinosinusitis: A Comparative Study in China.
BACKGROUND This study aimed to investigate the efficacy and safety of 3 nasal packing products (silicone tube, Beschitin-F, and Aquacel1-Ag) after functional endoscopic sinus surgery (FESS) for chronic rhinosinusitis (CRS) patients in China. MATERIAL AND METHODS Sixty-six CRS patients undergoing FESS surgery were randomly divided into 3 groups (22 patients in each group): the silicone tube group, the Beschitin-F group, and the Aquacel1-Ag group. Postoperative headache, nasal pain, nasal bleeding, and swelling of the nasal mucosa were observed at 1 month after FESS surgery. Bacteriology of chronic rhinosinusitis was conducted by culturing the removed nasal packing. RESULTS The VAS scores of nasal pain in the silicone tube group were lower than in the Beschitin-F and the Aquacel1-Ag groups. The volume of nasal bleeding in the Beschitin-F group was higher than in the silicone tube and Aquacel1-Ag groups. The adhesion rate of the nasal cavity in the Beschitin-F1 group (2/22, 9.1%) was also higher than in the silicone tube group (0/22, 0%) and the Aquacel1-Ag group (0/22, 0%). The results of bacterial culture from removed nasal packing showed that coagulase-negative staphylococci (CNS) was more frequent in the silicone tube group than in the Beschitin-F and Aquacel1-Ag groups, but Streptococcus pneumonia, Haemophilus influenza, and Gram-negative rods were more common in the Aquacel1-Ag group than in the silicone tube and Beschitin-F groups. CONCLUSIONS These results indicate that the silicone tube may be more effective and safe than Beschitin-F and Aquacel1-Ag as nasal packing after FES surgery for CRS.


Positive Culture Prior to Transperineal Prostate Biopsy Was Not Associated with Post-Biopsy Febrile Urinary Tract Infection Development.
To investigate the association between urine culture before transperineal prostate biopsy and post-biopsy febrile urinary tract infection (fUTI). We retrospectively reviewed 307 patients who underwent urine culture before transperineal prostate biopsy between April 2017 and September 2020. Patients with indwelling urinary catheters (n=7) were excluded. Urine culture was performed 1-3 days before the biopsy, and all patients received prophylactic cefazolin regardless of culture results. A urine culture was defined as positive if cell density was more than 1×105 colony-forming units per mL. Baseline characteristics and the incidence of post-biopsy fUTI were compared between patients showing positive pre-biopsy culture results and those showing negative findings. Out of 300, seven patients (2.3%) had positive urine culture results before the biopsy. Age (p=0.077); prostate-specific antigen at diagnosis (p=0.267); prostate volume (p=0.78); number of biopsy cores (p=0.277); percentage of patients testing positive for cancer on biopsy (p=0.71); and percentages of patients with a history of biopsy (p>0.999), diabetes mellitus (p=0.604), and immunosuppressive medication use (p>0.999) were similar between the two groups. No patient in the positive urine culture group had post-biopsy fUTI. However, 1.7% (five patients) of the negative urine culture group had the disease (p>0.999) (four patients with prostatitis and one with pyelonephritis). Among them, two patients were diagnosed by urine culture at the time of post-biopsy fUTI. In asymptomatic patients, positive pre-biopsy cultures were not associated with the development of post-biopsy fUTI.


Social network size relates to developmental neural sensitivity to biological motion.
The ability to perceive others' actions and goals from human motion (i.e., biological motion perception) is a critical component of social perception and may be linked to the development of real-world social relationships. Adult research demonstrates two key nodes of the brain's biological motion perception system-amygdala and posterior superior temporal sulcus (pSTS)-are linked to variability in social network properties. The relation between social perception and social network properties, however, has not yet been investigated in middle childhood-a time when individual differences in social experiences and social perception are growing. The aims of this study were to (1) replicate past work showing amygdala and pSTS sensitivity to biological motion in middle childhood; (2) examine age-related changes in the neural sensitivity for biological motion, and (3) determine whether neural sensitivity for biological motion relates to social network characteristics in children. Consistent with past work, we demonstrate a significant relation between social network size and neural sensitivity for biological motion in left pSTS, but do not find age-related change in biological motion perception. This finding offers evidence for the interplay between real-world social experiences and functional brain development and has important implications for understanding disorders of atypical social experience.


What European gynaecologists need to master: Consensus on medical expertise outcomes of pan-European postgraduate training in obstetrics & gynaecology.
European harmonisation of training standards in postgraduate medical education in Obstetrics and Gynaecology is needed because of the increasing mobility of medical specialists. Harmonisation of training will provide quality assurance of training and promote high quality care throughout Europe. Pan-European training standards should describe medical expertise outcomes that are required from the European gynaecologist. This paper reports on consensus development on the medical expertise outcomes of pan-European training in Obstetrics and Gynaecology. A Delphi procedure was performed amongst European gynaecologists and trainees in Obstetrics & Gynaecology, to develop consensus on outcomes of training. The consensus procedure consisted of two questionnaire rounds, followed by a consensus meeting. To ensure reasonability and feasibility for implementation of the training standards in Europe, implications of the outcomes were considered in a working group thereafter. We invited 142 gynaecologists and trainees in Obstetrics & Gynaecology for participation representing a wide range of European countries. They were selected through the European Board & College of Obstetrics and Gynaecology and the European Network of Trainees in Obstetrics & Gynaecology. Sixty people participated in round 1 and 2 of the consensus procedure, 38 (63.3%) of whom were gynaecologists and 22 (36.7%) were trainees in Obstetrics & Gynaecology. Twenty-eight European countries were represented in this response. Round 3 of the consensus procedure was performed in a consensus meeting with six experts. Implications of the training outcomes were discussed in a working group meeting, to ensure reasonability and feasibility of the material for implementation in Europe. The entire consensus procedure resulted in a core content of training standards of 188 outcomes, categorised in ten topics. European consensus was developed regarding the medical expertise outcomes of pan-European training in Obstetrics and Gynaecology. The outcomes will be described in core trainings standards, aimed at harmonising training in Obstetrics and Gynaecology in Europe to promote high quality care.


Dietary caffeine intake and bone status of postmenopausal women.
Dietary caffeine intake has been suggested as a risk factor for bone loss in postmenopausal women. We measured the bone density of both hips and the total body in 138 healthy, postmenopausal women aged 55-70 y who had either never used hormone replacement therapy (HRT) or had used HRT for < 1 y. In this cross-sectional study, participants were stratified according to their reported current and long-time caffeinated beverage use into one of three groups: low [0-2 cups (180 mL, or 6 oz per cup) caffeinated coffee per day], moderate (3-4 cups caffeinated coffee per day), or high (> or = 5 cups caffeinated coffee per day). Caffeine intake was measured from diet records and by gas chromatography of each subject's brewed, caffeinated beverages. No association between caffeine intake and any bone measurement was observed. The anthropometric and nutrient intakes of the three groups were similar. Compared with caffeine intake based on chemical analysis of brewed beverages, 3-d prospective food records and computer-assisted analysis overestimated caffeine intake by nearly two-thirds. In conclusion, the habitual dietary caffeine intake of this cohort of 138 postmenopausal women ranged from 0-1400 mg/d and was not associated with total body or hip bone mineral density measurements. This study does not support the notion that caffeine is a risk factor for bone loss in healthy postmenopausal women.


Therapeutic potential of curcumin in digestive diseases.
Curcumin is a low-molecular-weight hydrophobic polyphenol that is extracted from turmeric, which possesses a wide range of biological properties including anti-inflammatory, anti-oxidant, anti-proliferative and anti-microbial activities. Despite its diverse targets and substantial safety, clinical applications of this molecule for digestive disorders have been largely limited to case series or small clinical trials. The poor bioavailability of curcumin is likely the major hurdle for its more widespread use in humans. However, complexation of curcumin into phytosomes has recently helped to bypass this problem, as it has been demonstrated that this new lecithin formulation enables increased absorption to a level 29-fold higher than that of traditional curcuminoid products. This allows us to achieve much greater tissue substance delivery using significantly lower doses of curcumin than have been used in past clinical studies. As curcumin has already been shown to provide good therapeutic results in some small studies of both inflammatory and neoplastic bowel disorders, it is reasonable to anticipate an even greater efficacy with the advent of this new technology, which remarkably improves its bioavailability. These features are very promising and may represent a novel and effective therapeutic approach to both functional and organic digestive diseases.


Preparation and pharmacological evaluation of the R- and S-enantiomers of 3-(2-butylamino)-4H- and 3-(3-methyl-2-butylamino)-4H-pyrido[4,3-e]-1,2,4-thiadiazine 1,1-dioxide, two tissue selective ATP-sensitive potassium channel openers.
The preparation and the pharmacological evaluation of the R- and S-isomers of 3-(2-butylamino)-4H-pyrido[4,3-e]-1,2,4-thiadiazine 1,1-dioxide (BPDZ 42) and 3-(3-methyl-2-butylamino)-4H-pyrido[4,3-e]-1,2,4-thiadiazine 1,1-dioxide (BPDZ 44), two potassium channel openers, is described. Their optical purity was estimated by means of capillary electrophoresis (R- and S-BPDZ 42) and chiral HPLC (R- and S-BPDZ 44). The absolute configuration of each isomer of BPDZ 44 was deduced from crystallographic data. Pharmacological assays performed with the R- and S-isomers of BPDZ 44 revealed only slight differences in their activity on pancreatic B-cells but significant differences in their activity on vascular smooth muscle cells: the R-isomer being sixfold more potent than its corresponding S-isomer. The R-isomer of BPDZ 42 was shown to be more potent than its corresponding S-isomer on the endocrine pancreas. S-BPDZ 44 as well as R- and S-BPDZ 42 were found to exhibit tissue selectivity for the pancreatic versus the vascular smooth muscle tissue.


Disaggregation of Cause-Specific Cardiovascular Disease Mortality Among Hispanic Subgroups.
Hispanics are the largest minority group in the United States and face a disproportionate burden of risk factors for cardiovascular disease (CVD) and low socioeconomic position. However, Hispanics paradoxically experience lower all-cause mortality rates compared with their non-Hispanic white (NHW) counterparts. This phenomenon has been largely observed in Mexicans, and whether this holds true for other Hispanic subgroups or whether these favorable trends persist over time remains unknown. To disaggregate a decade of national CVD mortality data for the 3 largest US Hispanic subgroups. Deaths from CVD for the 3 largest US Hispanic subgroups-Mexicans, Puerto Ricans, and Cubans-compared with NHWs were extracted from the US National Center for Health Statistics mortality records using the underlying cause of death based on coding from the International Statistical Classification of Diseases and Related Health Problems, Tenth Revision (I00-II69). Mortality data were evaluated from January 1, 2003, to December 31, 2012. Population estimates were calculated using linear interpolation from the 2000 and 2010 US Census reports. Data were analyzed from November 2015 to July 2016. Mortality due to CVD. Participants included 688 074 Mexican, 163 335 Puerto Rican, 130 397 Cuban, and 19 357 160 NHW individuals (49.0% men and 51.0% women; mean [SD] age, 75 [15] years). At the time of CVD death, Mexicans (age, 67 [18] years) and Puerto Ricans (age, 68 [17] years) were younger compared with NHWs (age, 76 [15] years). Mortality rates due to CVD decreased from a mean of 414.2 per 100 000 in 2003 to 303.3 per 100 000 in 2012. Estimated decreases in mortality rate for CVD from 2003 to 2012 ranged from 85 per 100 000 for all Hispanic women to 144 per 100 000 for Cuban men, but rate differences between groups vary substantially, with Puerto Ricans exhibiting similar mortality patterns to NHWs, and Mexicans experiencing lower mortality. Puerto Ricans experienced higher mortality rates for ischemic and hypertensive heart disease compared with other subgroups, whereas Mexicans experienced higher rates of cerebrovascular disease deaths. Significant differences in CVD mortality rates and changes over time were found among the 3 largest Hispanic subgroups in the United States. Findings suggest that the current aggregate classification of Hispanics masks heterogeneity in CVD mortality reporting, leading to an incomplete understanding of health risks and outcomes in this population.


Reproducibility of glucose measurements using the glucose sensor.
Recent studies have confirmed that improved glycemic control decreases the risk of diabetic complications in type 1 and type 2 diabetic patients. The Minimed glucose sensor allows continuous 72-h glucose monitoring and represents a potentially important tool to improve diabetes management. Its use is currently limited to the health care team. Our aim was to evaluate the reproducibility of data provided by the device by comparing data provided by two sensors worn simultaneously by the same subject. A total of 11 subjects (6 type 1 and 3 type 2 diabetic patients and 2 healthy subjects) agreed to wear two sensors and perform at least four daily finger-stick glucose determinations during 72 h. The simultaneous glucose values provided by the sensors were compared. To determine the clinical implications of the glucose data, each day was divided into eight periods, and for each period the glucose range was rated as satisfactory, too high, or too low by a blinded clinician experienced in interpreting glucose sensor data in the clinical setting. The evaluation of glycemic levels based on the recordings of the two sensors were compared for each paired time interval. We discarded 18% of the sensor data for technical reasons. Examined as a group, the remaining 3,370 paired data points in all 11 patients were highly correlated (r = 0.84). However, when individual pairs were evaluated, large differences in the glucose values were apparent, with differences of >10% in 70% of the measurements and >50% in 7% of the measurements. Moreover, clinical evaluation of the glucose range provided simultaneously by two sensors was concordant for only 65% of the evaluation periods. In a real-life setting, the accuracy of data provided by the Minimed glucose sensor may be less than expected. To avoid therapeutic errors, sensor findings should be confirmed by independent means before clinical decisions are made.


Dengue risk assessment using multicriteria decision analysis: A case study of Bhutan.
Dengue is the most rapidly spreading vector-borne disease globally, with a 30-fold increase in global incidence over the last 50 years. In Bhutan, dengue incidence has been on the rise since 2004, with numerous outbreaks reported across the country. The aim of this study was to identify and map areas that are vulnerable to dengue in Bhutan. We conducted a multicriteria decision analysis (MCDA) using a weighted linear combination (WLC) to obtain a vulnerability map of dengue. Risk factors (criteria) were identified and assigned with membership values for vulnerability according to the available literature. Sensitivity analysis and validation of the model was conducted to improve the robustness and predictive ability of the map. Our study revealed marked differences in geographical vulnerability to dengue by location and season. Low-lying areas and those located along the southern border were consistently found to be at higher risk of dengue. The vulnerability extended to higher elevation areas including some areas in the Capital city Thimphu during the summer season. The higher risk was mostly associated with relatively high population density, agricultural and built-up landscapes and relatively good road connectivity. Using MCDA, our study identified vulnerable areas in Bhutan during specific seasons when and where the transmission of dengue is most likely to occur. This study provides evidence for the National Vector-borne Disease Control programme to optimize the use of limited public health resources for surveillance and vector control, to mitigate the public health threat of dengue.


Linezolid does not improve bactericidal activity of rifampin-containing first-line regimens in animal models of TB meningitis.
Tuberculous meningitis (TB meningitis) is the most devastating form of tuberculosis (TB) and there is a critical need to optimize treatment. Linezolid is approved for multidrug resistant TB and has shown encouraging results in retrospective TB meningitis studies, with several clinical trials underway assessing its additive effects on high-dose (35 mg/kg/day) or standard-dose (10 mg/kg/day) rifampin-containing regimens. However, the efficacy of adjunctive linezolid to rifampin-containing first-line TB meningitis regimens and the tissue pharmacokinetics (PK) in the central nervous system (CNS) are not known. We therefore conducted cross-species studies in two mammalian (rabbits and mice) models of TB meningitis to test the efficacy of linezolid when added to the first-line TB regimen and measure detailed tissue PK (multicompartmental positron emission tomography [PET] imaging and mass spectrometry). Addition of linezolid did not improve the bactericidal activity of the high-dose rifampin-containing regimen in either animal model. Moreover, the addition of linezolid to standard-dose rifampin in mice also did not improve its efficacy. Linezolid penetration (tissue/plasma) into the CNS was compartmentalized with lower than previously reported brain and cerebrospinal fluid (CSF) penetration, which decreased further two weeks after initiation of treatment. These results provide important data regarding the addition of linezolid for the treatment of TB meningitis.


Assay of human stem cells by repopulation of NOD/SCID mice.
The only conclusive method to assay stem cells is to follow their ability to repopulate conditioned recipients, making it difficult to study human stem cells. The development of systems to transplant human hematopoietic cells into immune-deficient mice lays the foundation for such an experimental repopulation assay for primitive human cells. Cell purification and gene marking studies have shown that the repopulating cells, termed severe-combined immunodeficiency (SCID) mouse-repopulating cells (SRC), are primitive and distinct from most of the progenitors that are detected using short and long-term in vitro culture assays. The SRC are exclusively CD34+CD38- and poorly infected with retrovirus vectors. These gene marking data are reminiscent of the human clinical trials establishing that the SRC assay is a good surrogate to develop improved transduction methods. Limiting dilution analysis has been used to establish a quantitative assay for SRC that can be used to precisely determine the effect of various cytokine cocktails on the proliferation and differentiation of SRC during in vitro culture.


Possible regulatory functions of protein kinase C-alpha and -epsilon isoenzymes in rat renal mesangial cells. Stimulation of prostaglandin synthesis and feedback inhibition of angiotensin II-stimulated phosphoinositide hydrolysis.
Short-term treatment of mesangial cells with phorbol 12-myristate 13-acetate (PMA) decreases angiotensin II-induced InsP3 formation, but potentiates hormone-stimulated arachidonic acid release and prostaglandin (PG) E2 synthesis. Long-term treatment with PMA augments hormone-stimulated InsP3 generation (after 8 h treatment), but eliminates angiotensin II-induced arachidonic acid release and PGE2 formation (after 24 h treatment). By using specific antibodies it is observed that mesangial cells express two protein kinase C (PKC) isoenzymes, PKC-alpha and -epsilon. No PKC-beta and -gamma isoenzymes are detected. On exposure to PMA a complete depletion of PKC-alpha is observed within 8 h. In contrast, down-regulation of PKC-epsilon to 10-20% of that found in control cells requires a 24 h treatment with PMA. These results may imply that PKC-alpha mediates feedback inhibition of phosphoinositide hydrolysis, whereas PKC-epsilon is a candidate for regulating PG synthesis in mesangial cells.


Personality, interests, and emotional disturbance in psychiatric residents.
To determine the personality and attitudinal characteristics of emotionally disturbed psychiatric residents and their short-term adjustment the authors conducted a series of psychological tests with physicians entering two psychiatric residency programs. Seventy residents completed the test battery, which included the General Health Questionnaire (GHQ), the MMPI, the Eysenck Personality Inventory, and the Strong Vocational Interest Blank. About 9% of the sample scored as cases on the GHQ, having nonpsychotic emotional illness, and a further 5% scored as likely cases. These physicians had significantly higher neuroticism scores and MMPI depression, social isolation, schizophrenia, psychasthenia, and hypochondriasis scores. Their interests were less like those of other physicians or psychiatrists and more like those of bankers and office workers.


Evaluation of functional integrity of the retinohypothalamic tract in advanced glaucoma using multifocal electroretinography and light-induced melatonin suppression.
The aim of the study was to investigate the survival of melanopsin-expressing retinal ganglion cells (mRGCs) and the functional integrity of the retinohypothalamic tract in patients with bilateral advanced glaucomatous optic neuropathy by measuring the neuroendocrine light response of the pineal gland. Nine patients with bilateral advanced primary open-angle glaucoma (glaucoma group) and nine normal control subjects (control group) were included in this pilot observational, prospective, case-control study. The best-corrected visual acuity logMAR, standard automated perimetry mean deviation, and the retinal nerve fiber layer thickness determined by optical coherence tomography and multifocal electroretinography were used to evaluate the changes. Melatonin was analyzed in the saliva by radioimmunoassay before and after exposure to bright light (600 lux) for 60 min at night. The advanced glaucoma group did not have any significant nocturnal melatonin suppression after exposure to bright light (14.28 ± 3.07 pg/ml pre-light melatonin concentration vs. 15.22 ± 3.56 pg/ml after light exposure; p = 0.798) unlike the marked melatonin suppression in the control group (22.43 ± 4.37 pg/ml pre-light melatonin concentration vs. 11.25 ± 1.89 pg/ml after light exposure; p < 0.002). Response density estimates by the scalar product amplitude measure for the interval 0-80 ms of the first-order kernel responses were similar in both groups, indicating that outer retinal function was significantly unchanged in the glaucoma group (5.95 ± 0.54 nV/dg^2) compared with the control group (6.20 ± 0.22 nV/dg^2) (p = 0.689). Our findings are consistent with the interpretation that the rhythmic secretion of melatonin was affected in advanced glaucoma, suggesting that attention should be paid to non-image-forming visual functions, such as control of circadian rhythm and the clinical impact in patients with glaucoma.


Rapid detection of biofilms and adherent pathogens using scanning confocal laser microscopy and episcopic differential interference contrast microscopy.
Knowledge of biofilm structure and function has changed significantly in the last few years due to advances in light microscopy. One pertinent example is the use of scanning confocal laser microscopy (SCLM) to visualise corrosion pits caused by the biofilm mosaic footprint on corroding metal surfaces. Nevertheless, SCLM has some limitations as to its widespread use, including cost, inability to observe motile bacteria and eukaryotic grazers within biofilms, and difficulty to scan a curved surface. By contrast, episcopic differential interference contrast (EDIC) microscopy has provided a rapid, real time analysis of biofilms on opaque, curved, natural or man-made surfaces without the need for cover slips and oil. EDIC, coupled with epi-fluorescence (EDIC/EF), microscopy has been used successfully to visualise the 3-D biofilm structure, physiological niches, protozoal grazing and iron biomineralization, and the location of specific pathogens such as Legionella pneumophila, Campylobacter jejuni and Cryptosporidium parvum. These species were identified using gold nanoparticles or fluorophores coupled to monoclonal antibodies or 16S rRNA probes, respectively. Among its many potential uses, the EDIC technique will provide a rapid procedure to facilitate the calibration of the modern generation of biofilm-sensing electrodes.


Antimicrobial and antiparasitic abietane diterpenoids from the roots of Clerodendrum eriophyllum.
Chromatographic separation of the roots of a Kenyan medicinal plant, Clerodendrum eriophyllum, led to the isolation of ten abietane diterpenoids (1-10), one of which (1) was isolated for the first time from a natural source. Using spectroscopic data, the structure of 1 was determined to be 12-hydroxy-8,12-abietadiene-3,11,14-trione. Circular dichroism (CD) spectra showed that the stereochemistry of compounds 1, 3, and 6-8 belongs to the normal series of abietane diterpenes, which confirmed the absolute stereochemistry of the isolated compounds. Compounds 1-10 were evaluated for their in vitro antiplasmodial, antileishmanial, antifungal and antibacterial activities. Compounds 3 and 7 exhibited potent antifungal activity (IC50/MIC 0.58/1.25 and 0.96/2.5 microg/mL, respectively) against C. neoformans, whereas 3, 6 and 7 showed strong antibacterial activity against Staphylococcus aureus and methicillin-resistant S. aureus with IC50/MIC values between 1.33-1.75/2.5-5 and 0.96-1.56/2.5 microg/mL, respectively. In addition, compounds 3 and 9 exhibited potent antileishmanial activity (IC50 0.08 and 0.20 microg/mL, respectively) against L. donovani, while 3 and 7 displayed weak antimalarial activity against Plasmodium falciparum, but 9 was inactive.


The influence of the intensity of smoking and years of work in the metallurgy on pro-oxidant/antioxidant balance in the blood of smelters.
The aim of this study was to investigate the effect of cigarette smoking and occupational exposure to heavy metals on the degree of pro-oxidant/antioxidant imbalance in smelters. The investigations were performed on the blood and urine of 400 subjects: 300 male copper smelters and 100 nonexposed male subjects. Biological material was divided into three groups: nonsmokers, those who smoked less than 20 cigarettes a day and those who smoked more than 20 cigarettes a day. The results showed a significant increase in the concentration of lead, cadmium and arsenic in the blood and urine of smelters, while smoking more than 20 cigarettes a day caused a further increase in the concentration of these metals. The level of malondialdehyde was approximately twofold higher in the plasma of the smelters compared to the control group. We have observed a disturbance in the level of antioxidants in erythrocyte lysate manifested by an increase in metallothionein and glutathione concentrations as well as superoxide dismutase and glutathione peroxidase activities and the decrease in glutathione S-transferase activity. Cigarette smoking, years of work in metallurgy and age of smelters were additional factors significantly affecting the pro-oxidant/antioxidant balance.


Biotransformation and improved enzymatic extraction of chlorogenic acid from coffee pulp by filamentous fungi.
The highest enzymatic extraction of covalent linked chlorogenic (36.1%) and caffeic (CA) (33%) acids from coffee pulp (CP) was achieved by solid-state fermentation with a mixture of three enzymatic extracts produced by Aspergillus tamarii, Rhizomucor pusillus, and Trametes sp. Enzyme extracts were produced in a practical inexpensive way. Synergistic effects on the extraction yield were observed when more than one enzyme extract was used. In addition, biotransformation of chlorogenic acid (ChA) by Aspergillus niger C23308 was studied. Equimolar transformation of ChA into CA and quinic acids (QA) was observed during the first 36 h in submerged culture. Subsequently, after 36 h, equimolar transformation of CA into protocatechuic acid was observed; this pathway is being reported for the first time for A. niger. QA was used as a carbon source by A. niger C23308. This study presents the potential of using CP to produce enzymes and compounds such as ChA with biological activities.


Leukocyte 12-lipoxygenase: expression, purification, and investigation of the role of methionine residues in turnover-dependent inactivation and 5,8,11,14-eicosatetraynoic acid inhibition.
Porcine leukocyte 12-lipoxygenase cDNA was cloned into the expression vectors pSE280, pSE380, and pSE420. pSE380 yielded the highest level of 12-lipoxygenase activity when these vectors were tested for expression in Escherichia coli Top10 cells. Optimal expression of the protein from this vector occurred in cells cultured at 30 degrees C and harvested 18 h following induction of expression by 0.5 mM isopropyl thiogalactoside (IPTG). The enzyme was purified from the 100000 g supernatant to 98% homogeneity by a combination of ammonium sulfate precipitation, anion exchange chromatography, and chromatofocusing. Addition of dithiothreitol and catalase to buffers at various steps in the purification protocol enabled the isolation of enzyme having a specific activity of 12 micromol min(-1) mg(-1). The recovery of purified protein from this expression system was 56%, resulting in a 109-fold purification. On the basis of amino acid sequence comparisons between mammalian 15- and 12-lipoxygenases, three methionine residues in the porcine leukocyte 12-lipoxygenase (M338L, M367V, and M562L) were targeted for mutation to assess their potential role in turnover-dependent inactivation and inhibition by 5,8,11,14-eicosatetraynoic acid (ETYA). The mutants were expressed and purified by the same procedure used for the wild-type enzyme. These amino acid changes did not significantly alter enzyme catalysis as judged by the kinetic constants Km and k(cat)/Km, nor did they affect the rate of turnover-dependent inactivation or inhibition by ETYA. The results indicate that these methionine residues do not play a pivotal role in catalysis, autoinactivation, or sensitivity to inhibition by acetylenic compounds.


The relative sensitivity of macrophyte and algal species to herbicides and fungicides: an analysis using species sensitivity distributions.
Lemna spp. are the standard test species representing aquatic macrophytes in the current risk assessment schemes for herbicides and plant growth regulators in the European Union and North America. At a Society of Environmental Toxicology and Chemistry (SETAC) 2008 workshop on Aquatic Macrophyte Risk Assessment for Pesticides (AMRAP), a Species Sensitivity Distribution (SSD) working group was formed to address uncertainties about the sensitivity of Lemna spp. relative to other aquatic macrophyte species. For 11 herbicides and 3 fungicides for which relevant and reliable data were found for at least 6 macrophyte species, SSDs were fitted using lognormal regression. The positions of L. gibba (the most commonly tested Lemna species) and Myriophyllum spicatum (for which standardized test methods are under development) in each SSD were determined where data were available. The sensitivity of standard algal test species required for pesticide registration in the United States under the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA) relative to the macrophytes in each SSD was also examined (algae were not included in the SSD). L. gibba was among the most sensitive macrophyte species for approximately 50% of the chemicals examined. M. spicatum was among the most sensitive macrophytes for approximately 25% of the chemicals. In most cases, the lowest FIFRA algal species endpoint was lower than the most sensitive macrophyte endpoint. Although no single species consistently represented the most sensitive aquatic plant species, for 12 of 14 chemicals L. gibba and the FIFRA algae included an endpoint near or below the 5th percentile of the macrophyte SSD. For the other compounds, M. spicatum was the most sensitive species of all aquatic plants considered.


Surgical Site Infection after Malignant Brain Tumor Resection: A Multicenter Study for Induction of a Basic Care Bundle.
Patients with malignant brain tumors are possibly at increased risk for surgical site infections (SSIs) considering the various medical situations associated with the disease. However, the actual rate of SSI after malignant brain tumor resection has not been well established, despite the potential impact of SSI on patient outcome. To investigate the incidence of SSI following malignant brain tumor surgery, we performed a retrospective study in 3 neurosurgical units. Subsequently, aiming at the reduction of incidence of SSI, we performed a prospective study using a care bundle technique in the same units. The SSI incidence in the retrospective (n = 161) and prospective studies (n = 68) were 4.3% and 4.4%, respectively, similar to the previously reports on general craniotomies. A care bundle does not appear to enhance prevention of SSI. However, future, large studies with a new care bundle should be planned based on a zero tolerance policy.


Control of adolescents' arbitrary matching-to-sample by positive and negative stimulus relations.
In Experiment 1, four developmentally delayed adolescents were taught an A-B matching-to-sample task with nonidentical stimuli: given Sample A1, select Comparison B1; given A2, select B2. During nonreinforced test trials, appropriate matching occurred when B stimuli appeared as samples and A stimuli as comparisons, i.e., the sample and comparison functions were symmetrical (B-A matching). During A-B or B-A matching test trials in which familiar samples and correct comparisons were presented along with novel comparisons, the subjects selected the correct comparisons. In tests with familiar samples and both incorrect and novel comparisons, subjects selected the novel comparisons, demonstrating control by both positive ("matching") and negative ("nonmatching") stimulus relations in A-B and B-A arrays. In Experiment 2, 12 developmentally delayed subjects were taught a two-stage arbitrary-matching task (e.g., A-B, C-B matching). Test sessions showed sample-comparison symmetry (e.g., B-A, B-C matching) and derived sample-comparison relations (e.g., A-C, C-A matching) for 11 subjects. These subjects also demonstrated control by positive and negative stimulus relations in the derived relations.


Anesthetic considerations for masticatory muscle tendon-aponeurosis hyperplasia: a report of 24 cases.
Masticatory muscle tendon-aponeurosis hyperplasia (MMTAH) is a new disease entity characterized by limited mouth opening due to contracture of the masticatory muscles, resulting from hyperplasia of tendons and aponeuroses. In this case series, we report what methods of airway establishment were conclusively chosen after rapid induction of anesthesia. We had 24 consecutive patients with MMTAH who underwent surgical release of its contracture under general anesthesia. Rapid induction of anesthesia with propofol and rocuronium was chosen for all the cases. In 7 cases, intubation using the Macintosh laryngoscopy was attempted; however, 2 of those cases failed to be intubated on the first attempt. Finally, intubation using the McCoy laryngoscopy or fiber-optic intubation was alternatively used in these 2 cases. In 7 cases, the Trachlight was used. In the remaining 10 cases, fiber-optic intubation was used. Limited mouth opening in patients with MMTAH did not improve with muscular relaxation. "Square mandible" has been reported to be one of the clinical features in this disease; however, half of these 24 patients lacked this characteristic, which might affect a definitive diagnosis of this disease for anesthesiologists. An airway problem in patients with MMTAH should not be underestimated, which means that other intubation methods rather than direct laryngoscopy had better be considered.


Systematic bias introduced by the informed consent process in a diagnostic research study.
To determine population characteristics, outcomes, and reasons for unsuccessful enrollment among potential study subjects approached for written, informed consent in a minimal-risk emergency department (ED) study. The authors hypothesized that the prevalence of venous thromboembolism (VTE) would be lower among study participants and that medical acuity and refusal to provide a blood sample would be the most common reasons for nonparticipation. The authors requested prospective, written, informed consent for a blood sample and follow-up from consecutive ED patients undergoing evaluation for pulmonary embolism (PE) and recorded spontaneously stated reasons for refusal. VTE was diagnosed or excluded using a combination of D-dimer testing and selective computed tomography (CT) angiography of the chest with venography of the lower extremities. The primary outcome was defined by the number of CT scans positive for VTE among ED patients evaluated for PE. Over 16 weeks, 260 of 287 (91%, 95% confidence interval [CI] = 87 to 94%) eligible patients were approached and consent was obtained from 183 patients (64%, 95% CI = 58% to 69%). The prevalence of VTE was 6% among participants and 13% among nonparticipants (95% CI [of the difference] = 1% to 15%). The proportions of African Americans, uninsured, and Medicaid patients were significantly higher among nonparticipants. No significant differences were found in the proportions of nonparticipants who disliked or distrusted research or desired financial reimbursement, compared to those not enrolled due to medical acuity or refused a blood sample. These data implicate the written, informed consent process as a significant source of bias on estimated disease prevalence.


Modifications of single acetylcholine-activated channels in BC3H-1 cells. Effects of trimethyloxonium and pH.
We have examined the effects of chemical modification with trimethyloxonium (TMO) and changes in external pH on the properties of acetylcholine (ACh)-activated channels in BC3H-1 cells, a clonal muscle cell line. TMO reacts covalently and specifically with carboxylic acid moieties in proteins to convert them to neutral methyl esters. In BC3H-1 cells TMO modification reduces the whole-cell response to ACh measured at negative membrane potentials by approximately 60%. G omega seal patch-clamp recordings of single ACh channel currents showed that the reduction in ACh sensitivity is due to alterations in both the current-carrying and the kinetic properties of the channels. Under all our experimental conditions, i.e., in external solutions of normal or low ionic strength, with or without external divalent cations, and at external pHs between 5.5 and 8.1, TMO treatment reduced ACh single-channel conductance to 70-90% of normal. The effects of TMO on channel kinetics were dependent on the ionic conditions. In normal ionic strength solutions containing both calcium and magnesium ions TMO modification reduced the channel average open time by approximately 25%. A similar reduction in open time was seen in calcium-free solution, but was not present when both calcium and magnesium ions were absent from the external solution. Lowering the ionic strength of the solution increased the mean open time in normal channels by about threefold, but did not affect the kinetics of modified channels. In low ionic strength solutions normal ACh channel open times were maximal at approximately pH 6.7 and decreased by three- to fourfold at both acid and alkaline pH. TMO modification removed the pH dependence of channel kinetics, and average open times were short at all pHs between 5.5 and 8.1. We suggest that TMO modifies normally titratable groups on the external surface of ACh channels that help to determine both the gating and permeability properties of ACh channels.


Deleterious functional impact of anemia on young adolescent school girls.
Iron deficiency anemia during adolescence may reduce physical work capacity and cognitive function. To assess the physical work capacity and cognition of underprivileged anemic schoolgirls in Vadodara in early adolescence as compared to their non-anemic counterparts. Prior to initiating anemia control interventions, schoolgirls in early adolescence were studied with regard to their hemoglobin status, physical work capacity and cognitive functions. Schoolgirls from four municipal primary schools in the age of 9 - 14 years were studied. In four Primary Municipal schools, hemoglobin of subjects was assessed using standard methods; physical work capacity using Modified Harvards Step test and cognitive functions using selected tests from the modified Wechsler Intelligence Scale for Children (WISC), suitably adapted for this group (n = 230). The mean hemoglobin was 11.32 g/dL, and anemia prevalence: 67%. A higher number of steps were climbed and a shorter time was taken to revert to the basal pulse rate (recovery time) by non-anemic girls compared to anemic girls (P < 0.001). Significantly lower scores in digit span and visual memory test were seen in anemic compared to non-anemic girls. The adverse impact of anemia remained after controlling for undernutrition (BMI). Anemia is likely to adversely affect physical work capacity and cognition in young adolescent girls undergoing pubertal development. Further research should be conducted in both school and community based settings to cover non-school going children.


Rotational acetabular osteotomy for secondary osteoarthritis after surgery for developmental dysplasia of the hip.
The treatment of residual deformity following surgery for developmental dysplasia of the hip remains controversial. The rationale for the use of the rotational acetabular osteotomy (RAO) is that it increases the weight-bearing area by shifting the osteotomized acetabulum to cover the femoral head. This can improve joint function as well as achieve relief of pain. However, it is unclear if this osteotomy can improve a compromised hip when performed for the treatment of residual deformity and acetabular dysplasia after surgery for developmental dysplasia of the hip. We aimed to report the clinical outcome as assessed by need for total hip arthroplasty (THA) and by the Merle d'Aubigné and Postel scores. In addition, we tried to assess the radiographic outcomes as assessed by Tönnis's classification. Only two hips required THA, which was performed in two patients at 11 and 12 years after RAO, respectively. The mean Merle d'Aubigné clinical score improved from 14.1 +/- 2.3 points (range, 10 to 17) preoperatively to 15.8 +/- 2.9 points (8 to 18) at final follow-up (p < 0.02). Radiological assessment at final follow-up showed the obvious progression of osteoarthritis in five hips. One patient in grade 1 preoperatively progressed into grade 3 at final follow-up; four patients in grade 2 preoperatively progressed into grade 3. In our study, this osteotomy prolonged the functional life of the hip, and only two hips needed THA after a mean follow-up of 11 years. We found that advanced arthritis pre-osteotomy is associated with progression of radiologic changes.


[Differences in the excitatory action of acetylcholine and mecholine on mechanosensitive high-threshold C-axonal units of the skin in the cat].
Intraarterial acetylcholine (Ach) and acetyl-beta-methylcholine (metacholine, Mch) excite feline high-threshold C-fiber mechanosensitive cutaneous sensory units (SU) in a different way. Ach in noxious (algogenic for humans) concentrations of 10 micrograms/ml and more induces a high-frequency discharge, with mean frequency of impulses at its peak being 4-12 Hz. This discharge seems to result from activation of N-cholinoreceptors because non-noxious and nonalgogenic M-agonist Mch as well as Ach in subnoxious concentrations induce in SU only a low-frequency discharge with a mean frequency of impulses 0.5-3.5 Hz. The data are discussed in connection with nociception and tissue chemoreception.


Total cholesterol and body mass index in relation to 40-year cancer mortality (the Corfu cohort of the seven countries study).
We evaluated risk factors of cancer mortality based on a 40-year follow-up of the Corfu cohort (Seven Countries Study). The population studied in this analysis consisted of 529 rural men (49 +/- 6 years old) enrolled in 1961. Since then, periodic visits every 5 years were made to define the causes of death of the participants. Cox proportional hazards models evaluated various risk factors in relation to cancer mortality. The death rate at the end of the follow-up was 87.1% (i.e., 461 deaths in 529 participants). Of those deaths, 118 (25.6%) were because of cancer (30 deaths were due to cancer of trachea, bronchus, and lung, and the rest were due to other malignant neoplasms). Cancer was the second cause of death in this cohort, after coronary heart disease. Age (hazard ratio, 1.05 per year; P < 0.05), smoking (hazard ratio, 1.97; P < 0.01), total serum cholesterol levels (hazard ratio, 0.95 per 10 mg/dL; P < 0.05), and body mass index (hazard ratio, 0.93 per 1 kg/m2; P < 0.05) showed a significant association with cancer deaths after controlling for physical activity status and anthropometric indices. It should be noted that the protective effect of total cholesterol on cancer mortality was observed only between 183 and 218 mg/dL baseline levels. Cancer was one of the leading causes of death in this cohort. Smoking was associated with increased risk of cancer, whereas moderate total serum cholesterol and increased body and mass index seemed to have a protective effect on 40-year cancer mortality.


Serologic response to human papillomavirus genotypes following vaccination: findings from the HITCH cohort study.
Human papillomavirus (HPV) infection contributes to approximately 5% of the worldwide cancer burden. The three-dose HPV vaccine has demonstrated immunogenicity and efficacy. Humoral responses may be critical for preventing, controlling, and/or eliminating HPV infection. Using data from the HITCH cohort, we analysed humoral immune response to HPV vaccination among women in relation to the phylogenetic relatedness of HPV genotypes. We included 96 women aged 18-24 years attending college or university in Montreal, Canada. Participants provided blood samples at enrolment and five follow-up visits. Antibody response to bacterially expressed L1 and E6 glutathione S-transferase fusion proteins of multiple Alphapapillomavirus types, and to virus-like particles (VLP-L1) of HPV16 and HPV18 were measured using multiplex serology. We assessed correlations between antibody seroreactivities using Pearson correlations (r). At enrolment, 87.7% of participants were unvaccinated, 2.4% had received one, 3.2% two, and 6.7% three doses of HPV vaccine. The corresponding L1 seropositivity to any HPV was 41.2%, 83.3%, 100%, and 97.0%. Between-type correlations for L1 seroreactivities increased with the number of vaccine doses, from one to three. Among the latter, the strongest correlations were observed for HPV58-HPV33 (Pearson correlation [r] = 0.96; α9-species); HPV11-HPV6 (r = 0.96; α10-species); HPV45-HPV18 (r = 0.95; α7-species), and HPV68-HPV59 (r = 0.95; α7-species). Correlations between HPV-specific antibody seroreactivities are affected by phylogenetic relatedness, with anti-L1 correlations becoming stronger with the number of vaccine doses received.