Arthroscopic agreement among surgeons on anterior cruciate ligament tunnel placement.
Little is known about surgeon agreement and accuracy using arthroscopic evaluation of anterior cruciate ligament (ACL) tunnel positioning. To investigate agreement on ACL tunnel position evaluated arthroscopically between operating surgeons and reviewing surgeons. We hypothesized that operating and evaluating surgeons would characterize tunnel positions significantly differently. Controlled laboratory study. Twelve surgeons drilled ACL tunnels on 72 cadaveric knees using transtibial (TT), medial portal (MP), or 2-incision (TI) techniques and then completed a detailed assessment form on tunnel positioning. Then, 3 independent blinded surgeon reviewers each arthroscopically evaluated tunnel position and completed the assessment form. Statistical comparisons of tunnel position evaluation between operating and reviewing surgeons were completed. Three-dimensional (3D) computed tomography (CT) scans were performed and compared with arthroscopic assessments. Arthroscopic assessments were compared with CT tunnel location criteria. Operating surgeons were significantly more likely to evaluate femoral tunnel position (92.6% vs 69.2%; P = .0054) and femoral back wall thickness as "ideal" compared with reviewing surgeons. Tunnels were judged ideal by reviewing surgeons more often when the TI technique was used compared with the MP and TT techniques. Operating surgeons were more likely to evaluate tibial tunnel position as ideal (95.5% vs 57.1%; P < .0001) and "acceptable" compared with reviewers. The ACL tunnels drilled using the TT technique were least likely to be judged as ideal on the tibia and the femur. Agreement among surgeons and observers was poor for all parameters (κ = -0.0053 to 0.2457). By 3D CT criteria, 88% of femoral tunnels and 78% of tibial tunnels were placed within applied criteria. Operating surgeons are more likely to judge their tunnels favorably than observers. However, independent surgeon reviewers appear to be more critical than results of 3D CT imaging measures. When subjectively evaluated arthroscopically, the TT technique yields more subjectively poorly positioned tunnels than the TI and MP techniques. Surgeons do not agree on the ideal placement for single-bundle ACL tunnels. This study demonstrates that surgeons do not currently uniformly agree on ideal single-bundle tunnel placement and that the TT technique may yield more poorly placed tunnels.


Dual Combined Real-Time Reverse Transcription Polymerase Chain Reaction Assay for the Diagnosis of Lyssavirus Infection.
The definitive diagnosis of lyssavirus infection (including rabies) in animals and humans is based on laboratory confirmation. The reference techniques for post-mortem rabies diagnosis are still based on direct immunofluorescence and virus isolation, but molecular techniques, such as polymerase chain reaction (PCR) based methods, are increasingly being used and now constitute the principal tools for diagnosing rabies in humans and for epidemiological analyses. However, it remains a key challenge to obtain relevant specificity and sensitivity with these techniques while ensuring that the genetic diversity of lyssaviruses does not compromise detection. We developed a dual combined real-time reverse transcription polymerase chain reaction (combo RT-qPCR) method for pan-lyssavirus detection. This method is based on two complementary technologies: a probe-based (TaqMan) RT-qPCR for detecting the RABV species (pan-RABV RT-qPCR) and a second reaction using an intercalating dye (SYBR Green) to detect other lyssavirus species (pan-lyssa RT-qPCR). The performance parameters of this combined assay were evaluated with a large panel of primary animal samples covering almost all the genetic variability encountered at the viral species level, and they extended to almost all lyssavirus species characterized to date. This method was also evaluated for the diagnosis of human rabies on 211 biological samples (positive n = 76 and negative n = 135) including saliva, skin and brain biopsies. It detected all 41 human cases of rabies tested and confirmed the sensitivity and the interest of skin biopsy (91.5%) and saliva (54%) samples for intra-vitam diagnosis of human rabies. Finally, this method was successfully implemented in two rabies reference laboratories in enzootic countries (Cambodia and Morocco). This combined RT-qPCR method constitutes a relevant, useful, validated tool for the diagnosis of rabies in both humans and animals, and represents a promising tool for lyssavirus surveillance.


Efficacy of electronic cigarettes for smoking cessation.
To review data demonstrating effective smoking cessation with electronic cigarettes (e-cigarettes). A literature search of MEDLINE/PubMed (1946-March 2014) was performed using the search terms e-cigarettes, electronic cigarettes, and smoking cessation. Additional references were identified from a review of literature citations. All English-language clinical studies assessing efficacy of e-cigarettes compared with baseline, placebo, or other pharmacological methods to aid in withdrawal symptoms, smoking reduction, or cessation were evaluated. A total of 6 clinical studies were included in the review. In small studies, e-cigarettes significantly decreased desire to smoke, number of cigarettes smoked per day, and exhaled carbon monoxide levels. Symptoms of nicotine withdrawal and adverse effects were variable. The most common adverse effects were nausea, headache, cough, and mouth/throat irritation. Compared with nicotine patches, e-cigarettes were associated with fewer adverse effects and higher adherence. Most studies showed a significant decrease in cigarette use acutely; however, long-term cessation was not sustained at 6 months. There is limited evidence for the effectiveness of e-cigarettes in smoking cessation; however, there may be a place in therapy to help modify smoking habits or reduce the number of cigarettes smoked. Studies available provided different administration patterns such as use while smoking, instead of smoking, or as needed. Short-term studies reviewed were small and did not necessarily evaluate cessation with a focus on parameters associated with cessation withdrawal symptoms. Though long-term safety is unknown, concerns regarding increased poisoning exposures among adults in comparison with cigarettes are alarming.


A regulatory network controls nephrocan expression and midgut patterning.
Although many regulatory networks involved in defining definitive endoderm have been identified, the mechanisms through which these networks interact to pattern the endoderm are less well understood. To explore the mechanisms involved in midgut patterning, we dissected the transcriptional regulatory elements of nephrocan (Nepn), the earliest known midgut specific gene in mice. We observed that Nepn expression is dramatically reduced in Sox17(-/-) and Raldh2(-/-) embryos compared with wild-type embryos. We further show that Nepn is directly regulated by Sox17 and the retinoic acid (RA) receptor via two enhancer elements located upstream of the gene. Moreover, Nepn expression is modulated by Activin signaling, with high levels inhibiting and low levels enhancing RA-dependent expression. In Foxh1(-/-) embryos in which Nodal signaling is reduced, the Nepn expression domain is expanded into the anterior gut region, confirming that Nodal signaling can modulate its expression in vivo. Together, Sox17 is required for Nepn expression in the definitive endoderm, while RA signaling restricts expression to the midgut region. A balance of Nodal/Activin signaling regulates the anterior boundary of the midgut expression domain.


Prevalent presence of periodic actin-spectrin-based membrane skeleton in a broad range of neuronal cell types and animal species.
Actin, spectrin, and associated molecules form a periodic, submembrane cytoskeleton in the axons of neurons. For a better understanding of this membrane-associated periodic skeleton (MPS), it is important to address how prevalent this structure is in different neuronal types, different subcellular compartments, and across different animal species. Here, we investigated the organization of spectrin in a variety of neuronal- and glial-cell types. We observed the presence of MPS in all of the tested neuronal types cultured from mouse central and peripheral nervous systems, including excitatory and inhibitory neurons from several brain regions, as well as sensory and motor neurons. Quantitative analyses show that MPS is preferentially formed in axons in all neuronal types tested here: Spectrin shows a long-range, periodic distribution throughout all axons but appears periodic only in a small fraction of dendrites, typically in the form of isolated patches in subregions of these dendrites. As in dendrites, we also observed patches of periodic spectrin structures in a small fraction of glial-cell processes in four types of glial cells cultured from rodent tissues. Interestingly, despite its strong presence in the axonal shaft, MPS is disrupted in most presynaptic boutons but is present in an appreciable fraction of dendritic spine necks, including some projecting from dendrites where such a periodic structure is not observed in the shaft. Finally, we found that spectrin is capable of adopting a similar periodic organization in neurons of a variety of animal species, including Caenorhabditis elegans, Drosophila, Gallus gallus, Mus musculus, and Homo sapiens.


Antimicrobial activity of Lactobacillus fermentum TcUESC01 against Streptococcus mutans UA159.
Dental caries is a multifactorial chronic-infection disease, which starts with a bacterial biofilm formation caused mainly by Streptococcus mutans. The use of probiotics has shown numerous health benefits, including in the fight against oral diseases. Strains of Lactobacillus fermentum have already shown probiotic potential against S. mutans, but there are still few studies. Thus, the aim of our study was to evaluate the antimicrobial activity of the inoculum and metabolites produced by L. fermentum TcUESC01 against S. mutans UA159. For this, a growth curve of L. fermentum was performed and both the inoculum and the metabolites formed in the fermentation were tested against the growth of S. mutans UA159 in agar diffusion tests, and only its metabolites were tested to determine the minimum inhibitory concentration, minimal bactericidal concentration and inhibition of cell adhesion. Inhibition of biofilm formation, pH drop and proton permeability were also tested with the metabolites. The zone of inhibition began to be formed at 14 h and continued until 16 h. The inoculum containing L. fermentum also showed zone of inhibition. The MIC for the metabolites was 1280 mg/mL and the MBC was obtained with a concentration higher than the MIC equal to 5120 mg/mL. Half of the MIC concentration (640 mg/mL) was required to inhibit S. mutans adhesion to the surface of the microplates. In the biofilm analyzes, the treatment with the metabolites in the tested concentration was not able to reduce biomass, insoluble glucans and alkali soluble compared to the control biofilm (p > 0.05). The metabolites also did not affect acid production and acid tolerance of S. mutans cells in biofilms compared to saline group (p > 0.05). Lactic acid (50.38%) was the most abundant organic acid produced by L. fermentum. This is the first report showing that the metabolites produced by the Lactobacillus fermentum TcUESC01 have a potential to be used as an antimicrobial agent against S. mutans, showing anti-adherence and bactericidal activity against planktonic cells of S. mutans. Thus, further studies should be carried out in order to better understand the antimicrobial activity of metabolites of L. fermentum TCUESC01.


Clinical outcomes after the endovascular treatments of pulmonary vein stenosis in patients with congenital heart disease.
Pulmonary vein stenosis (PVS) is a condition with challenging treatment and leads to severe cardiac failure and pulmonary hypertension. Despite aggressive surgical or catheter-based intervention, the prognosis of PVS is unsatisfactory. This study aimed to assess the prognosis and to establish appropriate treatment strategies. We retrospectively reviewed endovascular treatments for PVS (2001-2017) from the clinical database at the Okayama University Hospital. A total of 24 patients underwent PVS associated with total anomalous pulmonary venous connection and 7 patients underwent isolated congenital PVS. In total, 53 stenotic pulmonary veins were subjected to endovascular treatments; 40 of them were stented by hybrid (29) and percutaneous procedures (11) (bare-metal stent, n = 34; drug-eluting stent, n = 9). Stent size of hybrid stenting was larger than percutaneous stenting. Median follow-up duration from the onset of PVS was 24 months (4-134 months). Survival rate was 71 and 49% at 1 and 5 years, respectively. There was no statistically significant difference between stent placement and survival; however, patients who underwent bare-metal stent implantation had statistically better survival than those who underwent drug-eluting stent implantation or balloon angioplasty. Early onset of stenosis, timing of stenting, and small vessel diameter of pulmonary vein before stenting were considered as risk factors for in-stent restenosis. Freedom from re-intervention was 50 and 26% at 1 and 2 years. To improve survival and stent patency, implantation of large stent is important. However, re-intervention after stenting is also significant to obtain good outcome.


Camphor Sulfonic Acid Doped Poly(3-hexylthiophene) Nanofilms: Optical and Electrical Properties.
Herein, we report the fabrication and characterization of Camphor Sulfonic Acid (CSA) doped Poly(3-hexylthiophene) (P3HT) nanofilms prepared at different substrate temperature on glass by simple wire bar coating method. Spectroscopic, optic, structural and electrical properties of the prepared nanofilms were characterized by atomic force microscopy (AFM), UV-Visible spectroscopy, fluorescence spectroscopy, X-ray diffraction, NKD spectroscopy, SEM and DC electrical measurements. The detailed electrical characterizations revealed that the conductivity of the P3HT nanofilms increased when the films are doped with CSA and the conductivity increased from 5.89×10⁻⁵ S/cm to 1.39 × 10⁻⁴ S/cm for 40 °C preparation temperature. Thus, it was confirmed that the substrate temperature plays an important role on the film structure and spectroscopic properties. Additionally CSA-doping changes the optical properties, especially when the films prepared at 40 °C.


Viral Restriction Activity of Feline BST2 Is Independent of Its N-Glycosylation and Induction of NF-κB Activation.
BST2 (CD317, tetherin, HM1.24) is an interferon-inducible transmembrane protein which can directly inhibit the release of enveloped virus particles from infected cells, and its anti-viral activity is reported to be related to the specific topological arrangement of its four structural domains. The N-terminal cytoplasmic tail of feline BST2 (fBST2) is characterized by a shorter N-terminal region compared to those of other known homologs. In this study, we investigated the functional impact of modifying the cytoplasmic tail region of fBST2 and its molecular mechanism. The fBST2 protein with the addition of a peptide at the N-terminus retained anti-release activity against human immunodeficiency virus type-1 and pseudovirus based on feline immunodeficiency virus at a weaker level compared with the wild-type fBST2. However, the fBST2 protein with addition of a peptide internally in the ectodomain proximal to the GPI anchor still retained its anti-viral activity well. Notably, the N-glycosylation state and the cell surface level of the N-terminally modified variants were unlike those of the wild-type protein, while no difference was observed in their intracellular localizations. However, in contrast to human BST2, the wild-type fBST2 did not show the ability to activate NF-κB. Consistent with previous reports, our findings showed that adding a peptide in the cytoplasmic tail region of fBST2 may influence its anti-viral activity. The shorter N-terminal cytoplasmic region of fBST2 compared with human BST2 did not apparently affect its anti-viral activity, which is independent of its N-glycosylation and ability to activate NF-κB.


The Gut: A Key to the Pathogenesis of Type 2 Diabetes?
In this communication we discuss the role of the gut for the development of type 2 diabetes mellitus (T2DM). Gastric emptying rates importantly determine postprandial glucose excursions and regulate postprandial secretion of the incretin hormones, glucose-dependent insulinotropic polypeptide (GIP), and glucagon-like peptide-1 (GLP-1). It thereby also determines their powerful, amplifying effect on glucose-induced insulin secretion and thus the ability of the body to regulate glucose disposal. Although disturbances in gastric emptying are not consistent findings in type 2 diabetes, the incretin system is seriously impaired, probably associated with insulin resistance and obesity. Both of the incretin hormones lose (part of) their insulinotropic activity resulting, together with (genetically) defective beta cell function, in the impaired postprandial insulin secretion of T2DM. In addition, glucagon responses are inappropriately increased and importantly contribute to both fasting and postprandial hyperglycemia. This may involve stimulation by GIP, but evidence also points to a role of circulating amino acids, which are elevated due to steatosis-induced impaired glucagon-mediated hepatic clearance, in line with recent work suggesting that the alpha cells and the liver are linked in a close, amino acid-mediated feedback circuit. Thus, the gut plays an important role in the development of T2DM spurred by overeating and defective beta cells.


Optical pulse propagation through clouds.
The cloud impulse response (spatial and temporal) to optical pulse propagation has been measured. Experimental data are reported for the radiance function, pulse stretching, and (the first published) delay time. The results have been confirmed by Monte Carlo modeling. A geometric scattering model is presented explaining the temporal results for the test conditions.


Reliability of spike timing in neocortical neurons.
It is not known whether the variability of neural activity in the cerebral cortex carries information or reflects noisy underlying mechanisms. In an examination of the reliability of spike generation using recordings from neurons in rat neocortical slices, the precision of spike timing was found to depend on stimulus transients. Constant stimuli led to imprecise spike trains, whereas stimuli with fluctuations resembling synaptic activity produced spike trains with timing reproducible to less than 1 millisecond. These data suggest a low intrinsic noise level in spike generation, which could allow cortical neurons to accurately transform synaptic input into spike sequences, supporting a possible role for spike timing in the processing of cortical information by the neocortex.


The influence of repeated pain stimulation on the emotional aspect of pain: a preliminary study in healthy volunteers.
Pain is a multidimensional experience with sensory-discriminative, cognitive-evaluative, and affective-motivational components. Emotional factors, such as unpleasantness or anxiety, are known to have influence on pain in humans. Repeated painful stimulation has been reported to reduce subjective pain intensity. Nevertheless, there is little evidence of the influence of such stimulation on the emotional factors of pain. The aim of this study was to evaluate the influence of repeated painful stimulation on the experience of unpleasantness and anxiety. Eight subjects (six females, two males) volunteered to participate in this study. Subjects received repeated painful stimulation for 3 consecutive days each instance lasting 6 seconds, 60 times per day, on the medial side of the forearm of the nondominant hand. We examined the following items to evaluate changes of responses to painful stimulation: pain thresholds, pain tolerance levels, pain intensities, unpleasantness, and anxiety. Furthermore, pain thresholds and pain tolerance levels were compared between different sites on the ipsilateral and contralateral forearms. No immediate or chronological changes in pain thresholds or pain tolerance levels were observed. Pain intensities were reduced significantly over the 3-day experimental period (P<0.05). On the other hand, there was no great change in unpleasantness during the 3-day period. Anxiety was increased significantly after the painful stimulation compared with that without the painful stimulation and before day 1 of the stimulation (P<0.05). These results suggest that repeated painful stimulation may result in habituation to pain intensities but not habituation to emotional factors.


Late toxicity in the randomized multicenter HYPRO trial for prostate cancer analyzed with automated treatment planning.
Assess to what extent the use of automated treatment planning would have reduced organ-at-risk dose delivery observed in the randomized HYPRO trial for prostate cancer, and estimate related toxicity reductions. Investigate to what extent improved plan quality for hypofractionation scheme as achieved with automated planning can potentially reduce observed enhanced toxicity for the investigated hypofractionation scheme to levels observed for conventional fractionation scheme. For 725 trial patients, VMAT plans were generated with an algorithm for automated multi-criterial plan generation (autoVMAT). All clinically delivered plans (CLINICAL), generated with commonly applied interactive trial-and-error planning were also available for the investigations. Analyses were based on dose-volume histograms (DVH) and predicted normal tissue complication probabilities (NTCP) for late gastrointestinal (GI) toxicity. Compared to CLINICAL, autoVMAT plans had similar or higher PTV coverage, while large and statistically significant OAR sparing was achieved. Mean doses in the rectum, anus and bladder were reduced by 7.8 ± 4.7 Gy, 7.9 ± 6.0 Gy and 4.2 ± 2.9 Gy, respectively (p < 0.001). NTCPs for late grade ≥2 GI toxicity, rectal bleeding and stool incontinence were reduced from 23.3 ± 9.1% to 19.7 ± 8.9%, from 9.7 ± 2.8% to 8.2 ± 2.8%, and from 16.8 ± 8.5% to 13.1 ± 7.2%, respectively (p < 0.001). Reductions in rectal bleeding NTCP were observed for all published Equivalent Uniform Dose volume parameters, n. AutoVMAT allowed hypofractionation with predicted toxicity similar to conventional fractionation with CLINICAL plans. Compared to CLINICAL, autoVMAT had superior plan quality, with meaningful NTCP reductions for both conventional fractionation and hypofractionation schemes. AutoVMAT plans might reduce toxicity for hypofractionation to levels that were clinically observed (and accepted) for conventional fractionation. This may be relevant when considering clinical use of the investigated hypofractionation schedule with relatively high fraction dose (3.4 Gy).


Point-of-admission hypoglycaemia among under-five Nigerian children with plasmodium falciparum malaria: prevalence and risk factors.
Hypoglycaemia is a well recognized complication of falciparum malaria in children but its diagnosis may be overlooked because all the clinical features may be mimicked by severe malaria. To determine the prevalence of hypoglycaemia at the point of hospital admission of under-fives with falciparum malaria and identify its risk factors in patients seen in a Nigerian secondary-health-care institution. During a 12-month period and at the point of hospital admission, venous blood sample was collected into an appropriate sample bottle (fluoride-oxalate bottle) from 502 children who were below 5 years of age with positive falciparum malaria parasitaemia. The blood sample was analysed using the glucose-oxidase method. The duration of illness, degree of parasitaemia and time of last meal were noted for each child. Ninety two (18.3%) out of 502 children below five years old with falciparum malaria had hypoglycaemia ( blood glucose below 2.6 mmol/L or 50 mg/dl) at the point of hospital admission. Twenty three percent (78 out of 339) of children below 36 months old were hypoglycaemic compared to 8.6% (14 out of 163) children aged 36 months and above; (p=0.01). Prevalence of hypoglycaemia was higher in girls (20.7%) than boys (16.3%) [Odd ratio, OR=0.75 (95% Confidence Interval, CI=0.48-1.18)]. Forty (13.1%) out of 305 children whose time of last meal was 12 hours and below had hypoglycaemia compared to 52(26.4%) out of 197 whose time of last meal was greater than 12 hours; (p=0.02). Hypoglycaemia at admission point was associated with a significant increase in mortality rate; (p=0.00). The duration of illness and the degree of parasitaemia did not have significant difference with the prevalence of hypoglycaemia. In falciparum malaria, a greater interval (between 2 meals) than 12 hours in children below 36 months old predisposed them to hypoglycaemia. Routine monitoring of blood glucose at the admission point is suggested in malaria endemic region.


Requests for 692 transfers to an academic level I trauma center: implications of the emergency medical treatment and active labor act.
The Emergency Medical Treatment and Active Labor Act (EMTALA) effectively requires Level I trauma centers (TC) to accept all transfers for a higher level of care if capacity exists. We hypothesized that EMTALA would burden a Level I TC by a selective referral of a poor payer mix of primarily nonoperative patients. All transfer calls (December 2003 and September 2005) to our Level I TC are handled by a dedicated transfer center. Calls were reviewed for age, surgical service requested, and outcome of request. The trauma registry was queried to compare Injury Severity Scale (ISS) score, hospital stay (LOS), operations, mortality, and payer status for transfer and primary catchment patients. In all, 821 calls were received; 77 calls were cancelled by the referring hospital and 52 were for consultation only. Of the 692 transfer requests, 534 (77%) were accepted, 134 (19%) were denied for no capacity, and only 24 (4%) were declined by TC as not clinically indicated. Transferred patients were younger (32.0 +/- 1.49 versus 38.9 +/- 0.51, p < 0.05), had similar ISS scores (13.6 +/- 0.62 versus 13.7 +/- 0.26) and LOS (7.0 +/- 0.70 versus 7.4 +/- 0.25), but were somewhat more likely to require an operation than direct admissions (58% versus 51%, p < 0.05). Although trauma (24%) and neurosurgery (24%) were the most commonly requested services, followed by orthopedics (20%), orthopedics accounted for 60% of operations on transferred patients compared with 10% to 13% for trauma and neurosurgery (mostly spine). There was no difference in the payer status of transfer and direct admit patients. Contrary to our assumptions, EMTALA patients had an identical payer mix and similar operative need compared with our primary catchment patients. They do represent a large additional patient load (20% of admissions) and differentially impact specialists, mostly operative for orthopedics and complex nonoperative care for trauma and neurosurgery. These data suggest that the primary motivations for transfer are specialist availability and complexity of care rather than financial concerns. As TCs provide backup specialty call coverage for a wide geographic area, this further supports the need for trauma systems development.


Dopa, catecholamines and their metabolites in essential hypertension.
1. Urinary excretion of dopa, catecholamines and their metabolites (vanillylmandelic acid, methoxycatecholamines, 3-methoxy-4-hydroxyphenylglycol and homovanillic acid) were studied in eighty patients with essential hypertension and in twenty-five healthy control subjects. 2. Increased urinary excretion of catecholamines, dopa and catecholamine metabolites was found in a proportion of cases. 3. The relationship between urinary excretion of catecholamine metabolites and the excretion of dopa and noradrenaline was studied. 4. In view of the suggested significance of 3-methoxy-4-hydroxyphenylglycol as an index of brain catecholamine metabolism, particular attention was paid to urinary excretion of this metabolite in the subjects under study.


Validation of the revised Mystical Experience Questionnaire in experimental sessions with psilocybin.
The 30-item revised Mystical Experience Questionnaire (MEQ30) was previously developed within an online survey of mystical-type experiences occasioned by psilocybin-containing mushrooms. The rated experiences occurred on average eight years before completion of the questionnaire. The current paper validates the MEQ30 using data from experimental studies with controlled doses of psilocybin. Data were pooled and analyzed from five laboratory experiments in which participants (n=184) received a moderate to high oral dose of psilocybin (at least 20 mg/70 kg). Results of confirmatory factor analysis demonstrate the reliability and internal validity of the MEQ30. Structural equation models demonstrate the external and convergent validity of the MEQ30 by showing that latent variable scores on the MEQ30 positively predict persisting change in attitudes, behavior, and well-being attributed to experiences with psilocybin while controlling for the contribution of the participant-rated intensity of drug effects. These findings support the use of the MEQ30 as an efficient measure of individual mystical experiences. A method to score a "complete mystical experience" that was used in previous versions of the mystical experience questionnaire is validated in the MEQ30, and a stand-alone version of the MEQ30 is provided for use in future research.


Use of the prostate-specific antigen test among men aged 75 years or older in the United States: 2006 Behavioral Risk Factor Surveillance System.
In 2008, the US Preventive Services Task Force (USPSTF) updated prostate cancer screening guidelines to recommend against screening for prostate cancer in men aged 75 years or older. We describe the prevalence of prostate-specific antigen (PSA) testing in this population and identify factors that may be correlated with the use of this test. Data came from the 2006 Behavioral Risk Factor Surveillance System. We assessed the status of PSA testing in the past year among 9,033 US men aged 76 or older who had no history of prostate cancer. We conducted descriptive and multiple logistic regression analyses to assess associations of PSA testing with certain sociodemographic and psychosocial factors. Overall, 60% of men aged 76 or older reported having a PSA test in the past year. Men who had health insurance, were satisfied with life, or always had emotional support were significantly more likely to report having a PSA test in the past year. However, men who had no routine health checkup; were divorced, widowed, or separated; or had less than a high school education were significantly less likely to report having had a PSA test. PSA testing is common among men aged 75 or older in the United States. Certain sociodemographic and psychosocial factors were associated with receipt of this test. This study may not only provide baseline data to evaluate acceptance and implementation of the USPSTF screening guidelines but may also help physicians and public health providers better understand these sociodemographic and psychosocial factors in this population.


Normalization of mediotemporal and prefrontal activity, and mediotemporal-striatal connectivity, may underlie antipsychotic effects of cannabidiol in psychosis.
Recent evidence suggests that cannabidiol (CBD), a non-intoxicating ingredient present in cannabis extract, has an antipsychotic effect in people with established psychosis. However, the effect of CBD on the neurocognitive mechanisms underlying psychosis is unknown. Patients with established psychosis on standard antipsychotic treatment were studied on separate days at least one week apart, to investigate the effects of a single dose of orally administered CBD (600 mg) compared to a matched placebo (PLB), using a double-blind, randomized, PLB-controlled, repeated-measures, within-subject cross-over design. Three hours after taking the study drug participants were scanned using a block design functional magnetic resonance imaging (fMRI) paradigm, while performing a verbal paired associate learning task. Fifteen psychosis patients completed both study days, 13 completed both scanning sessions. Nineteen healthy controls (HC) were also scanned using the same fMRI paradigm under identical conditions, but without any drug administration. Effects of CBD on brain activation measured using the blood oxygen level-dependent hemodynamic response fMRI signal were studied in the mediotemporal, prefrontal, and striatal regions of interest. Compared to HC, psychosis patients under PLB had altered prefrontal activation during verbal encoding, as well as altered mediotemporal and prefrontal activation and greater mediotemporal-striatal functional connectivity during verbal recall. CBD attenuated dysfunction in these regions such that activation under its influence was intermediate between the PLB condition and HC. CBD also attenuated hippocampal-striatal functional connectivity and caused trend-level symptom reduction in psychosis patients. This suggests that normalization of mediotemporal and prefrontal dysfunction and mediotemporal-striatal functional connectivity may underlie the antipsychotic effects of CBD.


Smoldering multiple myeloma: when to observe and when to treat?
Smoldering multiple myeloma (SMM) is an asymptomatic disorder characterized by the presence of at least 3 g/dL of serum M-protein and/or 10% to 60% bone marrow plasma cell infiltration with no myeloma-defining event. The risk of progression to active multiple myeloma (MM) is not uniform and several markers are useful for identifying patients at high risk of progression. The definition of the disease has recently been revisited and patients with asymptomatic MM at 80% to 90% of progression risk at 2 years are now considered to have MM. Although the current standard of care is not to treat, a randomized trial in patients with high-risk SMM that compared early treatment versus observation demonstrated that early intervention resulted in substantial benefits in terms of time to progression and overall survival (OS). These findings highlight the need to follow a correct diagnosis by an accurate risk stratification to plan an optimized follow-up according to the risk of disease progression.


Intracavity electromagnetically induced transparency.
The effect of intracavity electromagnetically induced transparency (EIT) on the properties of optical resonators and active laser devices is discussed theoretically. Pronounced frequency pulling and cavity-linewidth narrowing are predicted. The EIT effect can be used to reduce classical and quantum-phase noise of the beat note of an optical oscillator substantially. Fundamental limits of this stabilization mechanism as well as its potential application to high-resolution spectroscopy are discussed.


Medical interpreting and the law in the European Union.
In 2011, the Danish government announced that from June that year it would no longer cover the costs of medical interpreters for patients who had been living in Denmark for more than seven years. The Dutch Ministry of Health followed with an even more draconian approach; from 1 January 2012, the cost of translation and interpreting would no longer be covered by the state. These two announcements led to widespread concern about whether or not there is a legal foundation for interpreter provision in healthcare. This article considers United Nations treaties, conventions from the Council of Europe and European Union law. European Union member states have been slow to sign up to international agreements to protect the rights of migrant workers. The European Union itself has only recently moved into the area of discrimination and it is unclear if the Race Directive covers language. As a result, access to interpreters in healthcare, where it exists, is dependent on national anti-discrimination legislation or on positive action taken at national or local level rather than on European or international law.


Further observations on luminal deformity and stenosis of nonrespiratory bronchioles in pulmonary emphysema.
In an endeavour to elucidate the anatomical basis for the increased resistance to airflow which characterises the most peripheral conducting air passages in pulmonary emphysema, lumina of nonrespiratory bronchioles of lungs with mainly centrilobular disease were assessed for two- and three-dimensional features by: (1) determination of percentage conformity of the lumina of individual bronchioles in histological sections to hypothetical planes through cylinders (ie, ellipses with the same areas and circumferences), and (2) comparison of luminal areas at regular intervals along bronchiolar longitudinal axes. The lumina of most nonrespiratory bronchioles from normal lungs conformed closely to their respective ellipses, thus corroborating previous observations that they are normally cylindroid. In contrast, there was a substantial excess of plane section deformities in the lumina of nonrespiratory bronchioles from the emphysematous specimens. The incidence of stenotic bronchioles (by both diameter and area determinations) was also greatly increased in emphysema. Since there was a strong positive association between such stenotic lesions and bronchiolar deformity, the latter was concluded to be a major factor in bronchiolar restriction. Furthermore, these characteristics seemed to have three-dimensional expression, for the lumina of stenotic bronchioles in emphysema were irregular in a longitudinal fashion.


Disruption of arterial perivascular drainage of amyloid-β from the brains of mice expressing the human APOE ε4 allele.
Failure of elimination of amyloid-β (Aβ) from the brain and vasculature appears to be a key factor in the etiology of sporadic Alzheimer's disease (AD) and cerebral amyloid angiopathy (CAA). In addition to age, possession of an apolipoprotein E (APOE) ε4 allele is a strong risk factor for the development of sporadic AD. The present study tested the hypothesis that possession of the APOE ε4 allele is associated with disruption of perivascular drainage of Aβ from the brain and with changes in cerebrovascular basement membrane protein levels. Targeted replacement (TR) mice expressing the human APOE3 (TRE3) or APOE4 (TRE4) genes and wildtype mice received intracerebral injections of human Aβ(40). Aβ(40) aggregated in peri-arterial drainage pathways in TRE4 mice, but not in TRE3 or wildtype mice. The number of Aβ deposits was significantly higher in the hippocampi of TRE4 mice than in the TRE3 mice, at both 3- and 16-months of age, suggesting that clearance of Aβ was disrupted in the brains of TRE4 mice. Immunocytochemical and Western blot analysis of vascular basement membrane proteins demonstrated significantly raised levels of collagen IV in 3-month-old TRE4 mice compared with TRE3 and wild type mice. In 16-month-old mice, collagen IV and laminin levels were unchanged between wild type and TRE3 mice, but were lower in TRE4 mice. The results of this study suggest that APOE4 may increase the risk for AD through disruption and impedance of perivascular drainage of soluble Aβ from the brain. This effect may be mediated, in part, by changes in age-related expression of basement membrane proteins in the cerebral vasculature.


Taxonomy and toxicity of Prorocentrum from Perhentian Islands (Malaysia), with a description of a non-toxigenic species Prorocentrum malayense sp. nov. (Dinophyceae).
Thirteen isolates of Prorocentrum species were established from the coral reefs of Perhentian Islands Marine Park, Malaysia and underwent morphological observations and molecular characterization. Six species were found: P. caipirignum, P. concavum, P. cf. emarginatum, P. lima, P. mexicanum and a new morphotype, herein designated as P. malayense sp. nov. Prorocentrum malayense, a species closely related to P. leve, P. cf. foraminosum, P. sp. aff. foraminossum, and P. concavum (Clade A sensu Chomérat et al. 2018), is distinguished from its congeners as having larger thecal pore size and a more deeply excavated V-shaped periflagellar area. Platelet arrangement in the periflagellar area of P. malayense is unique, with the presence of platelet 1a and 1b, platelet 2 being the most anterior platelet, and a broad calabash-shaped platelet 3. The species exhibits consistent genetic sequence divergences for the nuclear-encoded large subunit ribosomal RNA gene (LSU rDNA) and the second internal transcribed spacer (ITS2). The phylogenetic inferences further confirmed that it represents an independent lineage, closely related to species in Clade A sensu Chomérat et al. Pairwise comparison of ITS2 transcripts with its closest relatives revealed the presence of compensatory base changes (CBCs). Toxicity analysis showed detectable levels of okadaic acid in P. lima (1.0-1.6 pg cell-1) and P. caipirignum (3.1 pg cell-1); this is the first report of toxigenic P. caipirignum in the Southeast Asian region. Other Prorocentrum species tested, including the new species, however, were below the detection limit.


The social skills and attachment to dogs of children with autism spectrum disorder.
Children with Autism Spectrum Disorder (ASD) have deficits in social skills, and interaction with service dogs has been associated with increased social skills for children with ASD. In this telephone survey of 70 parents of children with ASD, children owning dogs had greater Mean scores for social skills, using the Social Skills Improvement System Rating Scale, while those with some type of pet (not excluding dogs) had significantly greater skills for subscale item "assertion". Parents described their children as attached to their dogs. Children owning dogs completed the Companion Animal Bonding Scale, and reported strong bonding with dogs. These findings suggest children with ASD may bond with their dogs, and pet ownership may be associated with increased social skills.


Computational superoscillation imaging beyond the Rayleigh limit from far-field measurements.
Far-field imaging beyond the Rayleigh limit is one of the most important challenges in optics, microwave, and ultrasonics. We propose a novel sparsity-promoted super-oscillation imaging scheme for reconstructing more universal objects in subwavelength scales, which solves a weighted optimization problem constrained by lp-norm-based sparsity regularization (0≤p≤1). We demonstrate numerically that the proposed imaging technique improves the resolution related to existing approaches remarkably for the case of very high signal-to-noise ratio (SNR), including the traditional super-oscillation imaging and sparsity-based super-resolution imaging. The standard superoscillation based super-resolution imaging approach can be regarded as the first-iteration solution of the proposed scheme. Numerical results for one- and two-dimensional super-resolution imaging are presented for validation.


Secretagogue-induced redistributions of Na,K-ATPase in rat lacrimal acini.
Lacrimal acinar cells secrete macromolecular products in an approximately isotonic, sodium chloride (NaCl)-rich fluid. The mechanisms of macromolecular product secretion depend in part on a recycling traffic of membrane constituents between the Golgi complex and the apical plasma membrane. In contrast, the acinar cell's mechanisms for secreting Na+ and Cl- depend largely on the fluxes of these ions through transporters expressed in the apical and basal-lateral membranes. In addition to accelerating the recycling of secretory vesicle membrane constituents, the cholinergic agonist carbachol also triggers a net redistribution of sodium potassium adenosine triphosphatase (Na,K-ATPase) ion pumps between Golgi-associated pools and the basal-lateral plasma membranes (Yiu SC, et al: J Membrane Biol 102:185, 1988). In the present study, acinar preparations from rat lacrimal glands were stimulated with either carbachol, epinephrine, or isoproterenol. All three agonist stimulated release of the secretory protein lactoperoxidase, but only carbachol significantly accelerated Na+ undirectional influx. Subcellular fractionation analyses of resting and stimulated preparations indicated that carbachol caused a significant translocation of Na,K-ATPase activity from a Golgi-associated compartment to the basal-lateral plasma membranes. Neither adrenergic agonist significantly increased the basal-lateral membrane Na,K-ATPase activity, but each triggered a distinct pattern of redistributions of Na,K-ATPase and the Golgi membrane marker, galactosyltransferase. The carbachol-induced augmentation of basal-lateral membrane Na,K-ATPase activity represents a mechanism by which the cell might compensate for increased Na+ influx.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of copper treatment on the composition and function of the bacterial community in the sponge Haliclona cymaeformis.
Marine sponges are the most primitive metazoan and host symbiotic microorganisms. They are crucial components of the marine ecological system and play an essential role in pelagic processes. Copper pollution is currently a widespread problem and poses a threat to marine organisms. Here, we examined the effects of copper treatment on the composition of the sponge-associated bacterial community and the genetic features that facilitate the survival of enriched bacteria under copper stress. The 16S rRNA gene sequencing results showed that the sponge Haliclona cymaeformis harbored symbiotic sulfur-oxidizing Ectothiorhodospiraceae and photosynthetic Cyanobacteria as dominant species. However, these autotrophic bacteria decreased substantially after treatment with a high copper concentration, which enriched for a heterotrophic-bacterium-dominated community. Metagenomic comparison revealed a varied profile of functional genes and enriched functions, including bacterial motility and chemotaxis, extracellular polysaccharide and capsule synthesis, virulence-associated genes, and genes involved in cell signaling and regulation, suggesting short-period mechanisms of the enriched bacterial community for surviving copper stress in the microenvironment of the sponge. Microscopic observation and comparison revealed dynamic bacterial aggregation within the matrix and lysis of sponge cells. The bacteriophage community was also enriched, and the complete genome of a dominant phage was determined, implying that a lytic phage cycle was stimulated by the high copper concentration. This study demonstrated a copper-induced shift in the composition of functional genes of the sponge-associated bacterial community, revealing the selective effect of copper treatment on the functions of the bacterial community in the microenvironment of the sponge. This study determined the bacterial community structure of the common sponge Haliclona cymaeformis and examined the effect of copper treatment on the community structure and functional gene composition, revealing that copper treatment had a selective effect on the functions of the bacterial community in the sponge. These findings suggest that copper pollution has an ecological impact on the sponge symbiont. The analysis showed that the untreated sponges hosted symbiotic autotrophic bacteria as dominant species, and the high-concentration copper treatment enriched for a heterotrophic bacterial community with enrichment for genes important for bacterial motility, supplementary cellular components, signaling and regulation, and virulence. Microscopic observation showed obvious bacterial aggregation and a reduction of sponge cell numbers in treated sponges, which suggested the formation of aggregates to reduce the copper concentration. The enrichment for functions of directional bacterial movement and supplementary cellular components and the formation of bacterial aggregates and phage enrichment are novel findings in sponge studies.


Factors associated with increasing rural doctor supply in Asia-Pacific LMICs: a scoping review.
More than 60% of the world's rural population live in the Asia-Pacific region. Of these, more than 90% reside in low- and middle-income countries (LMICs). Asia-Pacific LMICs rural populations are more impoverished and have poorer access to medical care, placing them at greater risk of poor health outcomes. Understanding factors associated with doctors working in rural areas is imperative in identifying effective strategies to improve rural medical workforce supply in Asia-Pacific LMICs. We performed a scoping review of peer-reviewed and grey literature from Asia-Pacific LMICs (1999 to 2019), searching major online databases and web-based resources. The literature was synthesized based on the World Health Organization Global Policy Recommendation categories for increasing access to rural health workers. Seventy-one articles from 12 LMICs were included. Most were about educational factors (82%), followed by personal and professional support (57%), financial incentives (45%), regulatory (20%), and health systems (13%). Rural background showed strong association with both rural preference and actual work in most studies. There was a paucity in literature on the effect of rural pathway in medical education such as rural-oriented curricula, rural clerkships and internship; however, when combined with other educational and regulatory interventions, they were effective. An additional area, atop of the WHO categories was identified, relating to health system factors, such as governance, health service organization and financing. Studies generally were of low quality-frequently overlooking potential confounding variables, such as respondents' demographic characteristics and career stage-and 39% did not clearly define 'rural'. This review is consistent with, and extends, most of the existing evidence on effective strategies to recruit and retain rural doctors while specifically informing the range of evidence within the Asia-Pacific LMIC context. Evidence, though confined to 12 countries, is drawn from 20 years' research about a wide range of factors that can be targeted to strengthen strategies to increase rural medical workforce supply in Asia-Pacific LMICs. Multi-faceted approaches were evident, including selecting more students into medical school with a rural background, increasing public-funded universities, in combination with rural-focused education and rural scholarships, workplace and rural living support and ensuring an appropriately financed rural health system. The review identifies the need for more studies in a broader range of Asia-Pacific countries, which expand on all strategy areas, define rural clearly, use multivariate analyses, and test how various strategies relate to doctor's career stages.


Proposing a Framework for the Restorative Effects of Nature through Conditioning: Conditioned Restoration Theory.
Natural environments have been shown to trigger psychological and physiological restoration in humans. A new framework regarding natural environments restorative properties is proposed. Conditioned restoration theory builds on a classical conditioning paradigm, postulating the occurrence of four stages: (i) unconditioned restoration, unconditioned positive affective responses reliably occur in a given environment (such as in a natural setting); (ii) restorative conditioning, the positive affective responses become conditioned to the environment; (iii) conditioned restoration, subsequent exposure to the environment, in the absence of the unconditioned stimulus, retrieves the same positive affective responses; and (iv) stimulus generalization, subsequent exposure to associated environmental cues retrieves the same positive affective responses. The process, hypothetically not unique to natural environments, involve the well-documented phenomenon of conditioning, retrieval, and association and relies on evaluative conditioning, classical conditioning, core affect, and conscious expectancy. Empirical findings showing that restoration can occur in non-natural environments and through various sensory stimuli, as well as findings demonstrating that previous negative experience with nature can subsequently lower restorative effects, are also presented in support of the theory. In integration with other existing theories, the theory should prove to be a valuable framework for future research.


Rotavirus A Genome Segments Show Distinct Segregation and Codon Usage Patterns.
Reassortment of the Rotavirus A (RVA) 11-segment dsRNA genome may generate new genome constellations that allow RVA to expand its host range or evade immune responses. Reassortment may also produce phylogenetic incongruities and weakly linked evolutionary histories across the 11 segments, obscuring reassortment-specific epistasis and changes in substitution rates. To determine the co-segregation patterns of RVA segments, we generated time-scaled phylogenetic trees for each of the 11 segments of 789 complete RVA genomes isolated from mammalian hosts and compared the segments' geodesic distances. We found that segments 4 (VP4) and 9 (VP7) occupied significantly different tree spaces from each other and from the rest of the genome. By contrast, segments 10 and 11 (NSP4 and NSP5/6) occupied nearly indistinguishable tree spaces, suggesting strong co-segregation. Host-species barriers appeared to vary by segment, with segment 9 (VP7) presenting the weakest association with host species. Bayesian Skyride plots were generated for each segment to compare relative genetic diversity among segments over time. All segments showed a dramatic decrease in diversity around 2007 coinciding with the introduction of RVA vaccines. To assess selection pressures, codon adaptation indices and relative codon deoptimization indices were calculated with respect to different host genomes. Codon usage varied by segment with segment 11 (NSP5) exhibiting significantly higher adaptation to host genomes. Furthermore, RVA codon usage patterns appeared optimized for expression in humans and birds relative to the other hosts examined, suggesting that translational efficiency is not a barrier in RVA zoonosis.


Vitamin E and cancer prevention: recent advances and future potentials.
Many animal and in vitro experiments have shown that the supplementation of diet with vitamin E within a certain dose range reduced the risk of chemical- and radiation-induced cancers. In vitro studies revealed that alpha-tocopheryl succinate (TS) induced differentiation and growth-inhibition in certain animal and human tumor cells in culture, whereas alpha-tocopherol (alpha-T), alpha-tocopheryl acetate (alpha-TA) and alpha-tocopheryl nicotinate (alpha-TN) were ineffective, alpha-TS also reduced basal and ligand-stimulated adenylate cyclase activity, and expression of c-myc and H-ras oncogenes in certain tumor cells in culture. The relative efficacy of various forms of vitamin E in cancer prevention in animal or human models has not been evaluated. Human epidemiologic studies utilizing retrospective and prospective case-control experimental designs are not suitable for evaluating the role of vitamin E in cancer prevention due to several inherent problems associated with these methodologies. Intervention trials utilizing vitamin E with appropriate biological and statistical rationales are most suitable for testing the role of vitamin E in cancer prevention in humans. Some human trials utilizing vitamin E alone or in combination with other nutrients are in progress.


Chronic cervical spinal cord injury and autonomic hyperreflexia in rats.
Although it is well established that patients with cervical spinal cord injury are prone to acute, marked, hypertensive episodes, i.e., autonomic hyperreflexia, the specific mechanisms mediating this sometimes-fatal phenomenon are not completely understood. In this report, we describe the preparation and characterization of a rat model of chronic cervical spinal cord injury and autonomic hyperreflexia. Adult male Sprague-Dawley rats were chronically instrumented with arterial, venous, and gastric catheters. Beginning the first day after a complete cervical spinal transection (CST) and continuing for 1 wk, acute hypertensive responses to a modest increase of urinary bladder pressure (0-20 mmHg) were studied. Mean arterial pressure increased 25.9 +/- 4.8 mmHg during bladder distension the first day after CST. This response was not significantly different 3, 5, and 7 days after CST (overall average = 18.0 +/- 2.3 mmHg). The pressor response to bladder distension was completely abolished by intravesical lidocaine and autonomic ganglionic blockade (atropine + hexamethonium). Responses to bladder distension were not observed after the administration of chloralose anesthesia. We conclude that after cervical spinal transection the rat exhibits autonomic hyperreflexia similar to that seen in humans with spinal injury. Furthermore, autonomic hyperreflexia is completely established within 24 h after CST in the rat. Finally, some spinal autonomic reflexes are suppressed by chloralose anesthesia in the rat.


Optical realization of bioinspired spiking neurons in the electron trapping material thin film.
A thin film of electron-trapping material (ETM), when combined with suitable optical bistability, is considered as a medium for optical implementation of bioinspired neural nets. The optical mechanism of ETM under blue light and near-infrared exposure has the inherent ability at the material level to mimic the crucial components of the stylized Hodgkin-Huxley model of biological neurons. Combining this unique property with the high-resolution capability of ETM, a dense network of bioinspired neurons can be realized in a thin film of this infrared stimulable storage phosphor. When combined with suitable optical bistability and optical interconnectivity, it has the potential of producing an artificial nonlinear excitable medium analog to cortical tissue.


Epidemiologic Measures for Quantifying the Incidence of Concussion in National Collegiate Athletic Association Sports.
Injury rates compare the relative frequency of sport-related concussions across groups. However, they may not be intuitive to policy makers, parents, or coaches in understanding the likelihood of concussion. To describe 4 measures of incidence (athlete-based rate, athlete-based risk, team-based rate, and team-based risk) during the 2011-2012 through 2014-2015 academic years. Descriptive epidemiology study. Aggregate injury and exposure data collected from the National Collegiate Athletic Association Injury Surveillance Program in 13 sports (men's baseball, basketball, football, ice hockey, lacrosse, soccer, and wrestling and women's basketball, ice hockey, lacrosse, soccer, softball, and volleyball). Collegiate student-athletes. Sport-related concussion data from the National Collegiate Athletic Association Injury Surveillance Program during the 2011-2012 through 2014-2015 academic years were analyzed. We calculated concussion rates per 1000 athlete-exposures (AEs), concussion risk, average number of concussions per team, and percentage of teams with at least 1 concussion. During the 2011-2012 through 2014-2015 academic years, 1485 concussions were sustained by 1410 student-athletes across 13 sports. Concussion rates ranged from 0.09/1000 AEs in men's baseball to 0.89/1000 AEs in men's wrestling. Concussion risk ranged from 0.74% in men's baseball to 7.92% in men's wrestling. The average ± SD number of concussions per team ranged from 0.25 ± 0.43 in men's baseball to 5.63 ± 5.36 in men's football. The percentage of teams with a concussion ranged from 24.5% in men's baseball to 80.6% in men's football. Although men's wrestling had a higher concussion rate and risk, men's football had the largest average number of concussions per team and the largest percentage of teams with at least 1 concussion. The risk of concussion, average number of concussions per team, and percentage of teams with concussions may be more intuitive measures of incidence for decision makers. Calculating these additional measures is feasible within existing injury surveillance programs, and this method can be applied to other injury types.


Attenuation and size distribution measurements of Definity and manipulated Definity populations.
The contrast agent Definity has recently been shown to have substantial nonlinear activity at high ultrasound frequencies (>10 MHz). In this study, measurements were performed to characterize the frequency dependent attenuation properties of Definity and populations of Definity that had been modified to preferentially isolate smaller bubbles through decantation or mechanical filtration. A narrowband pulse-echo substitution method was employed with a series of four transducers covering the frequency range from 2 to 50 MHz. "Native" Definity has peak in attenuation in the vicinity of 10 MHz and remains high until 50 MHz. This pattern is significantly different from other clinically approved agents and is consistent with recent reports of nonlinear activity at high frequencies. With increasing decantation times, the attenuation peak becomes more diffuse and occurs at progressively higher frequencies. By 3 h for example, attenuation continues to rise until 30 MHz. The bubble size distribution undergoes preferential skewing toward smaller bubbles with increasing decantation time. Between 30 s and 3 h, the mean bubble diameter goes from 3.99 to 0.98 micrometers. Mechanical filtration with 2 and 1 microm pores causes attenuation to rise until 15 and 40 MHz, respectively. Definity can therefore be manipulated to improve its relative activity at higher frequencies (>10 MHz), which has implications for ultrasound biomicroscopy and intravascular ultrasound applications. Further, these results suggest that agent handling can have a substantial impact on size distributions affecting lower frequency applications. Shell friction estimates derived from these data are lower than those reported for larger bubbles at lower frequencies.


Early childhood socioeconomic status is associated with circulating interleukin-6 among mid-life adults.
It is proposed that socioeconomic conditions in early childhood effect immune programming, with poorer conditions resulting in adult phenotypes that are prone to inflammation. Recent evidence supports this possibility, showing an inverse association of childhood SES with adult markers of systemic inflammation. In this study, we further investigate this association, extending prior studies to include an examination of multiple indices of SES across distinct periods of childhood. Subjects were 112 men and women, 40-60 years of age (88.6% Caucasian). Childhood SES was indexed by a composite of three indicators of parental wealth (parental home and vehicle ownership, and number of bedrooms per child in the family home) averaged across 2 year periods of childhood between 1 and 18 years old. Higher adult serum concentrations of interleukin (IL)-6 were associated with lower SES in early childhood (years 1-2) (β=-.05, p<.05), associations that were independent of adult age, personal income, educational attainment, gender, race, body mass index, and physical activity. These associations support recent suggestions that the early environment may program immune phenotypes that contribute to disease risk.


Autophagic Network Analysis of the Dual Effect of Sevoflurane on Neurons Associated with GABARAPL1 and 2.
Sevoflurane is commonly used as a general anesthetic in neonates to aged patients. Preconditioning or postconditioning with sevoflurane protects neurons from excitotoxic injury. Conversely, sevoflurane exposure induces neurotoxicity during early or late life. However, little is known about the underlying mechanism of the dual effect of sevoflurane on neurons. Autophagy is believed to control neuronal homeostasis. We hypothesized that autophagy determined the dual effect of sevoflurane on neurons. DTome was used to identify the direct protein target (DPT) of sevoflurane. The STRING database was employed to investigate the proteins associated with the DPTs. Protein-protein interaction was assessed using Cytoscape. WebGestalt was used to analyze gene set enrichment. The linkage between candidate genes and autophagy was identified using GeneCards. This study found that 23 essential DPTs of sevoflurane interacted with 77 proteins from the STRING database. GABARAPL1 and 2, both of which are DPT- and autophagy-associated proteins, were significantly expressed in the brain and enriched in GABAergic synapses. Taken together, our findings showed that the network of sevoflurane-DPT-GABARAPL1 and 2 is related to the dual effect of sevoflurane on neurons.


Elucidating the mechanism of ferrocytochrome c heme disruption by peroxidized cardiolipin.
The interaction of peroxidized cardiolipin with ferrocytochrome c induces two kinetically and chemically distinct processes. The first is a rapid oxidation of ferrocytochrome c, followed by a slower, irreversible disruption of heme c. The oxidation of ferrocytochrome c by peroxidized cardiolipin is explained by a Fenton-type reaction. Heme scission is a consequence of the radical-mediated reactions initiated by the interaction of ferric heme iron with peroxidized cardiolipin. Simultaneously with the heme c disruption, generation of hydroxyl radical is detected by EPR spectroscopy using the spin trapping technique. The resulting apocytochrome c sediments as a heterogeneous mixture of high aggregates, as judged by sedimentation analysis. Both the oxidative process and the destructive process were suppressed by nonionic detergents and/or high ionic strength. The mechanism for generating radicals and heme rupture is presented.


HOT or not: examining the basis of high-occupancy target regions.
High-occupancy target (HOT) regions are segments of the genome with unusually high number of transcription factor binding sites. These regions are observed in multiple species and thought to have biological importance due to high transcription factor occupancy. Furthermore, they coincide with house-keeping gene promoters and consequently associated genes are stably expressed across multiple cell types. Despite these features, HOT regions are solely defined using ChIP-seq experiments and shown to lack canonical motifs for transcription factors that are thought to be bound there. Although, ChIP-seq experiments are the golden standard for finding genome-wide binding sites of a protein, they are not noise free. Here, we show that HOT regions are likely to be ChIP-seq artifacts and they are similar to previously proposed 'hyper-ChIPable' regions. Using ChIP-seq data sets for knocked-out transcription factors, we demonstrate presence of false positive signals on HOT regions. We observe sequence characteristics and genomic features that are discriminatory of HOT regions, such as GC/CpG-rich k-mers, enrichment of RNA-DNA hybrids (R-loops) and DNA tertiary structures (G-quadruplex DNA). The artificial ChIP-seq enrichment on HOT regions could be associated to these discriminatory features. Furthermore, we propose strategies to deal with such artifacts for the future ChIP-seq studies.


A Transcriptomic Model of Postnatal Cardiac Effects of Prenatal Maternal Cortisol Excess in Sheep.
In utero treatment with glucocorticoids have been suggested to reprogram postnatal cardiovascular function and stress responsiveness. However, little is known about the effects of prenatal exposure to the natural corticosteroid, cortisol, on postnatal cardiovascular system or metabolism. We have demonstrated an increased incidence of stillbirth in sheep pregnancies in which there is mild maternal hypercortisolemia caused by infusion of 1 mg/kg/d cortisol. In order to model corticosteroid effects in the neonate, we created a second model in which cortisol was infused for 12 h per day for a daily infusion of 0.5 mg/kg/d. In this model we had previously found that neonatal plasma glucose was increased and plasma insulin was decreased compared to those in the control group, and that neonatal ponderal index and kidney weight were reduced and left ventricular wall thickness was increased in the 2 week old lamb. In this study, we have used transcriptomic modeling to better understand the programming effect of this maternal hypercortisolemia in these hearts. This is a time when both terminal differentiation and a shift in the metabolism of the heart from carbohydrates to lipid oxidation are thought to be complete. The transcriptomic model indicates suppression of genes in pathways for fatty acid and ketone production and upregulation of genes in pathways for angiogenesis in the epicardial adipose fat (EAT). The transcriptomic model indicates that RNA related pathways are overrepresented by downregulated genes, but ubiquitin-mediated proteolysis and protein targeting to the mitochondria are overrepresented by upregulated genes in the intraventricular septum (IVS) and left ventricle (LV). In IVS the AMPK pathway and adipocytokine signaling pathways were also modeled based on overrepresentation by downregulated genes. Peroxisomal activity is modeled as increased in EAT, but decreased in LV and IVS. Our results suggest that pathways for lipids as well as cell proliferation and cardiac remodeling have altered activity postnatally after the in utero cortisol exposure. Together, this model is consistent with the observed increase in cardiac wall thickness at necropsy and altered glucose metabolism observed in vivo, and predicts that in utero exposure to excess maternal cortisol will cause postnatal cardiac hypertrophy and altered responses to oxidative stress.


The relationship between hospital managers' leadership style and effectiveness with passing managerial training courses.
Background: Effective leadership that rises from managerial training courses is highly constructive in managing hospitals more effectively. This study aims at investigating the relationship between leadership effectiveness with providing management training courses for hospital managers. Methods: This was a cross-sectional study carried out on top and middle managers of 16 hospitals of Iran University of Medical Sciences. As a sample, 96 participants were selected through census method. Data were collected using leadership effectiveness and style questionnaire, whose validity and reliability were certified in previous studies. Pearson correlation coefficient and linear regressions were used for data analysis. Results: Leadership effectiveness score was estimated to be 4.36, showing a suitable status for managers' leadership effectiveness compared to the set criteria. No significant difference was found between leadership effectiveness and styles among managers who had passed the training courses with those who had not (p>0.05). Conclusion: Passing managerial training courses may have no significant effect on managers' leadership effectiveness, but there may be some other variables which should be meticulously studied.


Propafenone disposition and pharmacodynamics in normal and norepinephrine-induced cardiomyopathic rabbit hearts.
The myocardial disposition and pharmacodynamics of propafenone were studied in 10 normal and 10 norepinephrine-induced cardiomyopathic rabbit hearts. The left ventricular propafenone concentrations measured after perfusion of propafenone (0.3 microM) for 150 min were similar in the normal group (18 +/- 8 micrograms/g) compared to the cardiomyopathy group (20 +/- 5 micrograms/g, P = NS). However, the concentration of propafenone in cardiomyopathic left ventricular papillary muscle, which was always extensively involved in the inflammatory process, was significantly lower (11 +/- 2 micrograms/g) compared to normal papillary muscle (19 +/- 4 micrograms/g, P less than .05). During propafenone perfusion a significantly greater increment in ventricular conduction time was observed in the cardiomyopathy group (17 +/- 6 msec) compared to the normal group (12 +/- 3 msec, P less than .05). The propafenone myocardial concentration-effect relationships describing changes in QRS duration were shifted to the left in the cardiomyopathy group. Furthermore, the slopes of these linear concentration-effect relationships were greater in the cardiomyopathy group (1.80 +/- 0.60 msec/micrograms/g) compared to the normal group (1.07 +/- 0.25 msec/micrograms/g, P less than .01). The ventricular effective refractory period was shorter at base line in the cardiomyopathy hearts (156 +/- 21 msec) compared to the normal group (176 +/- 23 msec, P less than .08). However, propafenone effects on changes in the ventricular effective refractory period were similar in the two groups. Thus, the myocardial accumulation of propafenone is reduced in areas of extensive necrosis observed in norepinephrine-induced cardiomyopathy. As well, cardiomyopathic tissue is more responsive to propafenone effects on ventricular conduction time.


Matrix metalloproteinase 2 from human rheumatoid synovial fibroblasts. Purification and activation of the precursor and enzymic properties.
Human rheumatoid synovial cells in culture secrete at least three related metalloproteinases that digest extracellular matrix macromolecules. One of them, termed matrix metalloproteinase 2 (MMP-2), has been purified as an inactive zymogen (proMMP-2). The final product is homogeneous on SDS/PAGE with Mr = 72,000 under reducing conditions. The NH2-terminal sequence of proMMP-2 is Ala-Pro-Ser-Pro-Ile-Ile-Lys-Phe-Pro-Gly-Asp-Val-Ala-Pro-Lys-Thr, which is identical to that of the so-called '72-kDa type IV collagenase/gelatinase'. The zymogen can be rapidly activated by 4-aminophenylmercuric acetate to an active form of MMP-2 with Mr = 67,000, and the new NH2-terminal generated is Tyr-Asn-Phe-Phe-Pro-Arg-Lys-Pro-Lys-Trp-Asp-Lys-Asn-Gln-Ile. However, following 4-aminophenylmercuric acetate activation, MMP-2 is gradually inactivated by autolysis. Nine endopeptidases (trypsin, chymotrypsin, plasmin, plasma kallikrein, thrombin, neutrophil elastase, cathepsin G, matrix metalloproteinase 3, and thermolysin) were tested for their abilities to activate proMMP-2, but none had this ability. This contrasts with the proteolytic activation of proMMP-1 (procollagenase) and proMMP-3 (prostromelysin). The optimal activity of MMP-2 against azocoll is around pH 8.5, but about 50% of activity is retained at pH 6.5. Enzymic activity is inhibited by EDTA, 1,10-phenanthroline or tissue inhibitor of metalloproteinases, but not by inhibitors of serine, cysteine or aspartic proteinases. MMP-2 digests gelatin, fibronectin, laminin, and collagen type V, and to a lesser extent type IV collagen, cartilage proteoglycan and elastin. Comparative studies on digestion of collagen types IV and V by MMP-2 and MMP-3 (stromelysin) indicate that MMP-3 degrades type IV collagen more readily than MMP-2, while MMP-2 digests type V collagen effectively. Biosynthetic studies of MMPs using cultured human rheumatoid synovial fibroblasts indicated that the production of both proMMP-1 and proMMP-3 is negligible but it is greatly enhanced by the treatment with rabbit-macrophage-conditioned medium, whereas the synthesis of proMMP-2 is constitutively expressed by these cells and is not significantly affected by the treatment. This suggests that the physiological and/or pathological role of MMP-2 and its site of action may be different from those of MMP-1 and MMP-3.


Acetylation of MKP-1 and the control of inflammation.
Innate immune responses mediated by Toll-like receptors (TLRs), a class of pattern-recognition receptors, play a critical role in the defense against microbial pathogens. However, excessive TLR-mediated responses result in sepsis, autoimmunity, and chronic inflammation. To prevent deleterious activation of TLRs, cells have evolved multiple mechanisms that inhibit innate immune reactions. Stimulation of TLRs induces the expression of the gene encoding the mitogen-activated protein kinase (MAPK) phosphatase-1 (MKP-1), a nuclear-localized dual-specificity phosphatase that preferentially dephosphorylates p38 MAPK and c-Jun N-terminal kinase (JNK), resulting in the attenuation of TLR-triggered production of proinflammatory cytokines. MKP-1 is posttranslationally modified by multiple mechanisms, including phosphorylation. A study now demonstrates that MKP-1 is also acetylated on a key lysine residue following stimulation of TLRs. Acetylation of MKP-1 promotes the interaction of MKP-1 with its substrate p38 MAPK, which results in dephosphorylation of p38 MAPK and the inhibition of innate immunity.


Different influence of cardiac hemodynamics on thromboembolic events in patients with paroxysmal and non-paroxysmal atrial fibrillation.
Blood stasis in left atrium (LA) or LA appendage (LAA) is thought to be the main cause of thrombus formation and systemic embolism in atrial fibrillation (AF) patients. Paroxysmal and non-paroxysmal AF differ significantly in various aspects. Impact of cardiac hemodynamics on systemic embolism might also differ between the 2 distinct AF entities. This study was performed to evaluate the influence of cardiac hemodynamics on systemic embolism in both paroxysmal and non-paroxysmal AF. Consecutive AF patients undergoing radiofrequency catheter ablation (RFCA) in Korea University Medical Center Anam Hospital between June 1998 and February 2018 were analyzed. Among 2,801 patients who underwent first-time RFCA, a total of 231 patients had either previous ischemic stroke, transient ischemic attack, or arterial embolism. In paroxysmal AF, LA diameter, LA volume (measured with magnetic resonance imaging), left ventricular (LV) ejection fraction, E/e', LAA flow velocity, and prevalence of spontaneous echocontrast (SEC) and dense SEC were significantly different between patients with and without thromboembolic events. However, only E/e' was different between patients with and without thromboembolic events in non-paroxysmal AF. The influence of LA diameter, LA volume, LV EF, LAA flow velocity, and dense SEC on thromboembolic events was significantly moderated by the type of AF. In conclusion, paroxysmal and non-paroxysmal AF might have a different mechanism responsible for thrombus formation and consequent embolic events. Relative contribution of hemodynamic parameters and other factors such as atrial myopathy to thromboembolic events in paroxysmal versus non-paroxysmal AF needs further evaluation.


A steroidomic approach for biomarkers discovery in doping control.
Anti-doping authorities have high expectations of the athlete steroidal passport (ASP) for anabolic-androgenic steroids misuse detection. However, it is still limited to the monitoring of known well-established compounds and might greatly benefit from the discovery of new relevant biomarkers candidates. In this context, steroidomics opens the way to the untargeted simultaneous evaluation of a high number of compounds. Analytical platforms associating the performance of ultra-high pressure liquid chromatography (UHPLC) and the high mass-resolving power of quadrupole time-of-flight (QTOF) mass spectrometers are particularly adapted for such purpose. An untargeted steroidomic approach was proposed to analyse urine samples from a clinical trial for the discovery of relevant biomarkers of testosterone undecanoate oral intake. Automatic peak detection was performed and a filter of reference steroid metabolites mass-to-charge ratio (m/z) values was applied to the raw data to ensure the selection of a subset of steroid-related features. Chemometric tools were applied for the filtering and the analysis of UHPLC-QTOF-MS(E) data. Time kinetics could be assessed with N-way projections to latent structures discriminant analysis (N-PLS-DA) and a detection window was confirmed. Orthogonal projections to latent structures discriminant analysis (O-PLS-DA) classification models were evaluated in a second step to assess the predictive power of both known metabolites and unknown compounds. A shared and unique structure plot (SUS-plot) analysis was performed to select the most promising unknown candidates and receiver operating characteristic (ROC) curves were computed to assess specificity criteria applied in routine doping control. This approach underlined the pertinence to monitor both glucuronide and sulphate steroid conjugates and include them in the athletes passport, while promising biomarkers were also highlighted.


Ghost transmission: How large basis sets can make electron transport calculations worse.
The Landauer approach has proven to be an invaluable tool for calculating the electron transport properties of single molecules, especially when combined with a nonequilibrium Green's function approach and Kohn-Sham density functional theory. However, when using large nonorthogonal atom-centered basis sets, such as those common in quantum chemistry, one can find erroneous results if the Landauer approach is applied blindly. In fact, basis sets of triple-zeta quality or higher sometimes result in an artificially high transmission and possibly even qualitatively wrong conclusions regarding chemical trends. In these cases, transport persists when molecular atoms are replaced by basis functions alone ("ghost atoms"). The occurrence of such ghost transmission is correlated with low-energy virtual molecular orbitals of the central subsystem and may be interpreted as a biased and thus inaccurate description of vacuum transmission. An approximate practical correction scheme is to calculate the ghost transmission and subtract it from the full transmission. As a further consequence of this study, it is recommended that sensitive molecules be used for parameter studies, in particular those whose transmission functions show antiresonance features such as benzene-based systems connected to the electrodes in meta positions and other low-conducting systems such as alkanes and silanes.


Women's autonomy and experience of physical violence within marriage in rural India: evidence from a prospective study.
Evidence regarding the relationship between married women's autonomy and risk of marital violence remains mixed. Moreover, studies examining the contribution of specific aspects of women's autonomy in influencing the risk of marital violence using measures of autonomy that incorporate its dynamic nature are rare. We investigated the relationship between women's autonomy and their experience of marital violence in rural India using prospective data. We used data on 4,904 rural women drawn from two linked studies: the NFHS-2, conducted during 1998-1999 and a follow-up study for a subgroup of women carried out during 2002-2003. Three dimensions of autonomy were used: financial autonomy, freedom of movement, and household decision-making. Marital violence was measured as experience of physical violence in the year prior to the follow-up survey. Findings indicate the protective effects of financial autonomy and freedom of movement in reducing the risk of marital violence in the overall model. Furthermore, region-wise analysis revealed that in the more gender equitable settings of south India, financial autonomy exerted a protective influence on risk of marital violence. However, in the more gender-stratified settings of north India, none of the dimensions of autonomy were found to have any protective effect on women's risk of marital violence. Results argue for an increased focus on strategies aimed at improving women's financial status through livelihood skill-building opportunities, development of a strong savings orientation, and asset-building options.


Velnacrine for Alzheimer's disease.
Alzheimer's disease (AD) is the commonest cause of dementia affecting older people. One of the therapeutic strategies aimed at ameliorating the clinical manifestations of Alzheimer's disease is to enhance cholinergic neurotransmission in relevant parts of the brain by the use of cholinesterase inhibitors to delay the breakdown of acetylcholine released into synaptic clefts. Tacrine, the first of the cholinesterase inhibitors to undergo extensive trials for this purpose, was associated with significant adverse effects including hepatotoxicity. Velnacrine is a derivative of tacrine. To determine the clinical efficacy and safety of velnacrine for patients with dementia of Alzheimer's type. The trials were identified from a last updated search of the Specialized Register of the Cochrane Dementia and Cognitive Improvement Group on 8 January 2004 using the terms velnacr* and 'HP 029'. The CDCIG SR is regularly updated and contains records from all major health care databases and a great many ongoing trial databases. All unconfounded, double-blind, randomized trials in which treatment with velnacrine was administered to patients with dementia of the Alzheimer's type for more than two weeks and its effects compared with those of placebo in a parallel group of patients. One reviewer (JSB) applied study selection criteria, assessed the quality of studies and extracted data. Four trials, involving 899 participants, were included. Cutler 1990 reported only the adverse events, and omitted the results for the placebo group. Medication was stopped for all in the highest dose group after the fourth day because one member suffered a tonic seizure. It is not possible to report any comparisons with placebo. Antuono 1995 reported benefit for velnacrine at endpoint for the CGI-C, and the Physical Self-maintenance Scale (PGIR), but not for the Carers Assessment of Time Activity (CATS). These results could not be checked because the relevant information was not reported, and we cannot assess the effect on the results of the higher number of non-completers from the velnacrine group. Treatment was discontinued because of safety reasons in 135 patients, mostly due to an abnormal liver function tests. There was a significant difference in favour of placebo compared with the combined treatment group for the number with an abnormal liver function test before the end of treatment at 24 weeks [105/297 vs 4/152, OR =20.23, 95% CI 7.29 to 56.18, p<0.00001]. There was a significant difference in favour of placebo compared with the combined treatment group for the number of withdrawals before the end of treatment at 24 weeks [130/297 vs 39/152, OR =2.26, 95% CI 1.47 to 3.47, p=0.0002]. Results are available for the dose replication phases of Zemlan 1996a and Zemlan 1996b. All the patients had taken velnacrine within two weeks prior to this phase and were identified as responders to velnacrine defined by improvement on the ADAS-Cog. Both studies reported a significant benefit for velnacrine compared with placebo for the ADAS-cog, but the results could not be checked because the relevant information was not reported. Neither study reported any benefit for velnacrine for the other efficacy measures. There was a significant difference in favour of placebo compared with the treatment group for the number with elevated liver transaminases before the end of treatment at 6 weeks [45/153 vs 29/156, OR =1.82, 95% CI 1.07 to 3.11, p=0.03]. There was a significant difference in favour of placebo compared with the treatment group for the number of withdrawals before the end of treatment at 6 weeks [68/211 vs 47/215, OR =1.70, 95% CI 1.10 to 2.62, p=0.02]. There has been no research into the use of velnacrine as a cognitive enhancer in the treatment of Alzheimer's disease since 1994. The FDA peripheral and CNS drug advisory board voted unanimously against recommending approval. This review shows the toxic nature of velnacrine, and provides no evidence of efficacy. There are no grounds for further research into velnacrine.


Sensitivity of self-reported noncognitive skills to survey administration conditions.
Noncognitive skills (e.g., persistence and self-control) are typically measured using self-reported questionnaires in which respondents rate their own skills. In many applications-including program evaluation and school accountability systems-such reports are assumed to measure only the skill of interest. However, self-reports might also capture other dimensions aside from the skill, such as aspects of a respondent's situation, which could include incentives and the conditions in which they complete the questionnaire. To explore this possibility, this study conducted 2 experiments to estimate the extent to which survey administration conditions can affect student responses on noncognitive skill questionnaires. The first experiment tested whether providing information about the importance of noncognitive skills to students directly affects their responses, and the second experiment tested whether incentives tied to performance on another task indirectly affect responses. Both experiments suggest that self-reports of noncognitive skills are sensitive to survey conditions. The effects of the conditions are relatively large compared with those found in the program evaluation literature, ranging from 0.05 to 0.11 SDs. These findings suggest that the effects of interventions or other social policies on self-reported noncognitive skills should be interpreted with caution.


Regioselective magnesiation of N-heterocyclic molecules: securing insecure cyclic anions by a β-diketiminate-magnesium clamp.
Using a specially designed magnesium metallating manifold, combining kinetically activated TMP amide base with a sterically amplified β-diketiminate ligand, this study has established a new regioselective strategy for magnesiation of challenging N-heterocyclic molecules. The broad scope of the approach is illustrated through reactions of pyrazine, triazoles and substituted pyridines by isolation and structural elucidation of their magnesiated intermediates.


Detection of VIM-1-Producing Enterobacter cloacae and Salmonella enterica Serovars Infantis and Goldcoast at a Breeding Pig Farm in Germany in 2017 and Their Molecular Relationship to Former VIM-1-Producing S. Infantis Isolates in German Livestock Production.
In 2011, VIM-1-producing Salmonella enterica serovar Infantis and Escherichia coli were isolated for the first time in four German livestock farms. In 2015/2016, highly related isolates were identified in German pig production. This raised the issue of potential reservoirs for these isolates, the relation of their mobile genetic elements, and potential links between the different affected farms/facilities. In a piglet-producing farm suspicious for being linked to some blaVIM-1 findings in Germany, fecal and environmental samples were examined for the presence of carbapenemase-producing Enterobacteriaceae and Salmonella spp. Newly discovered isolates were subjected to Illumina whole-genome sequencing (WGS) and S1 pulsed-field gel electrophoresis (PFGE) hybridization experiments. WGS data of these isolates were compared with those for the previously isolated VIM-1-producing Salmonella Infantis isolates from pigs and poultry. Among 103 samples, one Salmonella Goldcoast isolate, one Salmonella Infantis isolate, and one Enterobacter cloacae isolate carrying the blaVIM-1 gene were detected. Comparative WGS analysis revealed that the blaVIM-1 gene was part of a particular Tn21-like transposable element in all isolates. It was located on IncHI2 (ST1) plasmids of ∼290 to 300 kb with a backbone highly similar (98 to 100%) to that of reference pSE15-SA01028. SNP analysis revealed a close relationship of all VIM-1-positive S Infantis isolates described since 2011. The findings of this study demonstrate that the occurrence of the blaVIM-1 gene in German livestock is restricted neither to a certain bacterial species nor to a certain Salmonella serovar but is linked to a particular Tn21-like transposable element located on transferable pSE15-SA01028-like IncHI2 (ST1) plasmids, being present in all of the investigated isolates from 2011 to 2017.IMPORTANCE Carbapenems are considered one of few remaining treatment options against multidrug-resistant Gram-negative pathogens in human clinical settings. The occurrence of carbapenemase-producing Enterobacteriaceae in livestock and food is a major public health concern. Particularly the occurrence of VIM-1-producing Salmonella Infantis in livestock farms is worrisome, as this zoonotic pathogen is one of the main causes for human salmonellosis in Europe. Investigations on the epidemiology of those carbapenemase-producing isolates and associated mobile genetic elements through an in-depth molecular characterization are indispensable to understand the transmission of carbapenemase-producing Enterobacteriaceae along the food chain and between different populations to develop strategies to prevent their further spread.


Memory T cell, exhaustion, and tumor immunity.
CD8+T cells are important in protective immunity against intracellular pathogens and tumors. In chronic infections or cancer, CD8+T cells are constantly exposed to antigens and inflammatory signals. Such excessive and constitutive signals lead to the deterioration of T cell function, called 'exhaustion'. Exhausted T cells are characterized by low proliferation in response to antigen stimulation, progressive loss of effector function (cytokine production and killing function), expression of multiple inhibitory receptors such as PD-1, Tim3, and LAG3, and metabolic alterations from oxidative phosphorylation to glycolysis. These dysfunctions are associated with altered transcriptional programs and epigenetic regulations and recent studies suggested that NR4a and TOX transcription factors are deeply involved in exhaustion phenotypes. However, an increase the early memory T cells including stem cell memory T (TSCM) cells is critical for T cell persistence and efficient tumor killing especially for adoptive cancer immunotherapy such as CAR-T cell therapy. An increasing amount of evidence supports the therapeutic potential of targeting exhausted T cells and TSCM cells. We have begun to understand the molecular mechanisms of T cell exhaustion and early memory formation, and the clinical application of converting exhausted T cells to rejuvenated early memory T cells is the goal of our study.


From structure to function: the ecology of host-associated microbial communities.
In the past several years, we have witnessed an increased interest in understanding the structure and function of the indigenous microbiota that inhabits the human body. It is hoped that this will yield novel insight into the role of these complex microbial communities in human health and disease. What is less appreciated is that this recent activity owes a great deal to the pioneering efforts of microbial ecologists who have been studying communities in non-host-associated environments. Interactions between environmental microbiologists and human microbiota researchers have already contributed to advances in our understanding of the human microbiome. We review the work that has led to these recent advances and illustrate some of the possible future directions for continued collaboration between these groups of researchers. We discuss how the application of ecological theory to the human-associated microbiota can lead us past descriptions of community structure and toward an understanding of the functions of the human microbiota. Such an approach may lead to a shift in the prevention and treatment of human diseases that involves conservation or restoration of the normal community structure and function of the host-associated microbiota.


Methods for data analysis.
The molecular epidemiology of infectious diseases uses a variety of techniques to assay the relatedness of disease-causing organisms to identify strains responsible for outbreaks or associated with particular phenotypes of interest (such as antibiotic resistance) and, it is hoped, provide insights into where and how these strains have emerged. The correct analysis of such data requires that we understand how the assayed variation accumulates. We discuss this with specific reference to three classes of methods: those based on gel electrophoresis of fragments generated by restriction enzymes or polymerase chain reaction (PCR), those based on microsatellites and other repeat elements, and raw sequence data from protein-coding genes. We also provide a simple example of how the likely origin of an apparently novel antibiotic-resistant strain may be identified and conclude with a discussion of some popular analysis packages and the more interesting prospects for the future in this rapidly developing field.


Pediatric Emergency Department Visits at US Children's Hospitals During the COVID-19 Pandemic.
The impact of the coronavirus disease 2019 (COVID-19) pandemic on pediatric emergency department (ED) visits is not well characterized. We aimed to describe the epidemiology of pediatric ED visits and resource use during the pandemic. We conducted a cross-sectional study using the Pediatric Health Information System for ED visits to 27 US children's hospitals during the COVID-19 pandemic period (March 15, 2020, to August 31, 2020) and a 3-year comparator period (March 15 to August 31, 2017-2019). ED visit rates, patient and visit characteristics, resource use, and ED charges were compared between the time periods. We specifically evaluated changes in low-resource-intensity visits, defined as ED visits that did not result in hospitalization or medication administration and for which no laboratory tests, diagnostic imaging, or procedures were performed. ED visit rates decreased by 45.7% (average 911 026 ED visits over 2017-2019 vs 495 052 visits in 2020) during the pandemic. The largest decrease occurred among visits for respiratory disorders (70.0%). The pandemic was associated with a relative increase in the proportion of visits for children with a chronic condition from 23.7% to 27.8% (P < .001). The proportion of low-resource-intensity visits decreased by 7.0 percentage points, and total charges decreased by 20.0% during the pandemic period. The COVID-19 pandemic was associated with a marked decrease in pediatric ED visits across a broad range of conditions; however, the proportional decline of poisoning and mental health visits was less pronounced. The impact of decreased visits on patient outcomes warrants further research.