Plaque formation by virulent Shigella flexneri.
An in vitro tissue culture plaque assay was developed to investigate the intracellular replication and intercellular spread of virulent shigellae. Shigella plaques were formed in HeLa cell monolayers in the presence of an agarose overlay containing tissue culture medium and gentamicin, which eliminated extracellular bacterial growth. Microscopically, the plaques were characterized by a central area of dead host cells surrounded by cells infected with shigellae. Cells further away from the plaque center were uninfected. Inclusion of chloramphenicol or nalidixic acid in the overlay completely abolished plaque formation. Plaque formation was completely inhibited when infected monolayers were shifted from 37 to 30 degrees C. Shifting infected monolayers from 30 degrees C, where plaques do not form, to 37 degrees C resulted in the formation of plaques. Cultures of Shigella boydii, Shigella sonnei (form I), and all six serotypes of Shigella flexneri produced plaques. Shigellae isolated from plaques were Sereny test positive, contained a 140-megadalton plasmid, and were gentamicin sensitive. Noninvasive shigellae did not form plaques.


Transmission of SARS-CoV-2 by inhalation of respiratory aerosol in the Skagit Valley Chorale superspreading event.
During the 2020 COVID-19 pandemic, an outbreak occurred following attendance of a symptomatic index case at a weekly rehearsal on 10 March of the Skagit Valley Chorale (SVC). After that rehearsal, 53 members of the SVC among 61 in attendance were confirmed or strongly suspected to have contracted COVID-19 and two died. Transmission by the aerosol route is likely; it appears unlikely that either fomite or ballistic droplet transmission could explain a substantial fraction of the cases. It is vital to identify features of cases such as this to better understand the factors that promote superspreading events. Based on a conditional assumption that transmission during this outbreak was dominated by inhalation of respiratory aerosol generated by one index case, we use the available evidence to infer the emission rate of aerosol infectious quanta. We explore how the risk of infection would vary with several influential factors: ventilation rate, duration of event, and deposition onto surfaces. The results indicate a best-estimate emission rate of 970 ± 390 quanta/h. Infection risk would be reduced by a factor of two by increasing the aerosol loss rate to 5 h-1 and shortening the event duration from 2.5 to 1 h.


Healthcare workers and immunity to infectious diseases.
In 2002, New South Wales (NSW) Health introduced an updated policy for occupational screening and vaccination against infectious diseases. This study describes healthcare worker (HCW) immunity to hepatitis B, measles, mumps, rubella (MMR) and varicella based on serological screening, following introduction of this policy. HCW screening serology performed at two healthcare facilities in south western Sydney (Bankstown and Fairfield) was extracted for the period September 2003 to September 2005. Immunity to hepatitis B, MMR and varicella was quantitated and cross-tabulated against age, sex and staff risk category. A total of 1,320 HCWs were screened. Almost two thirds were immune to hepatitis B while immunity to MMR and varicella ranged from 88% to 94%. Age stratification showed lower levels of measles immunity in those born after 1965. Despite availability of vaccination for over two decades, a significant proportion of HCWs at these two facilities were non-immune to hepatitis B. This is of concern for those non-immune staff involved in direct clinical care, who are at risk of blood and body fluid exposures. The small group of HCWs non-immune to MMR and varicella pose a risk to themselves and others in the event of an outbreak. There is a need for improved implementation of the occupational screening and vaccination policy, including better education of HCWs about the risks of non-immunity to vaccine preventable diseases. The revised 2007 NSW Health policy may assist this process and will need evaluation to determine whether HCW immunity improves in the coming years.


A mathematical model to predict endothelial cell density following penetrating keratoplasty with selective dropout from graft failure.
We constructed several mathematical models that predict endothelial cell density (ECD) for patients after penetrating keratoplasty (PK) for a moderate-risk condition (principally Fuchs' dystrophy or pseudophakic/aphakic corneal edema). In a subset (n = 591) of Cornea Donor Study participants, postoperative ECD was determined by a central reading center. Various statistical models were considered to estimate the ECD trend longitudinally over 10 years of follow-up. A biexponential model with and without a logarithm transformation was fit using the Gauss-Newton nonlinear least squares algorithm. To account for correlated data, a log-polynomial model was fit using the restricted maximum likelihood method. A sensitivity analysis for the potential bias due to selective dropout was performed using Bayesian analysis techniques. The three models using a logarithm transformation yield similar trends, whereas the model without the transform predicts higher ECD values. The adjustment for selective dropout turns out to be negligible. However, this is possibly due to the relatively low rate of graft failure in this cohort (19% at 10 years). Fuchs' dystrophy and pseudophakic/aphakic corneal edema (PACE) patients had similar ECD decay curves, with the PACE group having slightly higher cell densities by 10 years. Endothelial cell loss after PK can be modeled via a log-polynomial model, which accounts for the correlated data from repeated measures on the same subject. This model is not significantly affected by the selective dropout due to graft failure. Our findings warrant further study on how this may extend to ECD following endothelial keratoplasty.


Repeated, Close Physician Coronary Artery Bypass Grafting Teams Associated with Greater Teamwork.
To determine whether observed patterns of physician interaction around shared patients are associated with higher levels of teamwork as perceived by physicians. Michigan Medicare beneficiaries who underwent coronary artery bypass grafting (CABG) procedures at 24 hospitals in the state between 2008 and 2011. We assessed hospital teamwork using the teamwork climate scale in the Safety Attitudes Questionnaire. After aggregating across CABG discharges at these hospitals, we mapped the physician referral networks (including both surgeons and nonsurgeons) that served them and measured three network properties: (1) reinforcement, (2) clustering, and (3) density. We then used multilevel regression models to identify associations between network properties and teamwork at the hospitals on which the networks were anchored. In hospitals where physicians repeatedly cared for patients with the same colleagues, physicians perceived better teamwork (β-reinforcement = 3.28, p = .003). When physicians who worked together also had other colleagues in common, the reported teamwork was stronger (β clustering = 1.71, p = .001). Reported teamwork did not change when physicians worked with a higher proportion of other physicians at the hospital (β density = -0.58, p = .64). In networks with higher levels of reinforcement and clustering, physicians perceive stronger teamwork, perhaps because the strong ties between them create a shared understanding; however, sharing patients with more physicians overall (i.e., density) did not lead to stronger teamwork. Clinical and organizational leaders may consider designing the structure of clinical teams to increase interactions with known colleagues and repeated interactions between providers.


Hippocampal neurofibrillary tangle changes and aggressive behaviour in dementia.
Neuropsychiatric behaviours occur frequently in Alzheimer's disease and other dementias and are thought to arise from the neurodegenerative process. However, it is unclear whether neurodegenerative changes in the hippocampus are associated with neuropsychiatric behaviours such as aggression. In this study, semiquantitative measurements of cell loss, atrophy, neuritic plaque and neurofibrillary tangle load in the postmortem hippocampus were taken for dementia patients, prospectively assessed for neuropsychiatric behaviours. It was found that increased tangle load, but not other hippocampal neuropathological variables, was associated with increased severity of aggressive behaviours and presence of chronic aggression. This study suggests a pathogenic link between neurofibrillary tangle load and aggressive behaviours in the hippocampus of dementia patients.


Ptf1a-mediated control of Dll1 reveals an alternative to the lateral inhibition mechanism.
Neurog3-induced Dll1 expression in pancreatic endocrine progenitors ostensibly activates Hes1 expression via Notch and thereby represses Neurog3 and endocrine differentiation in neighboring cells by lateral inhibition. Here we show in mouse that Dll1 and Hes1 expression deviate during regionalization of early endoderm, and later during early pancreas morphogenesis. At that time, Ptf1a activates Dll1 in multipotent pancreatic progenitor cells (MPCs), and Hes1 expression becomes Dll1 dependent over a brief time window. Moreover, Dll1, Hes1 and Dll1/Hes1 mutant phenotypes diverge during organ regionalization, become congruent at early bud stages, and then diverge again at late bud stages. Persistent pancreatic hypoplasia in Dll1 mutants after eliminating Neurog3 expression and endocrine development, together with reduced proliferation of MPCs in both Dll1 and Hes1 mutants, reveals that the hypoplasia is caused by a growth defect rather than by progenitor depletion. Unexpectedly, we find that Hes1 is required to sustain Ptf1a expression, and in turn Dll1 expression in early MPCs. Our results show that Ptf1a-induced Dll1 expression stimulates MPC proliferation and pancreatic growth by maintaining Hes1 expression and Ptf1a protein levels.


A wireless electrochemiluminescence detector applied to direct and indirect detection for electrophoresis on a microfabricated glass device.
A novel electrochemiluminescence (ECL) detector is presented in this article. The detector is applied for micellar electrokinetic chromatographic separation of dichlorotris(2,2'-bipyridyl)ruthenium(II) hydrate [Ru-(bpy)] and dichlorotris(1,10-phenanthroline)ruthenium-(II) hydrate [Ru(phen)] on a microfabricated glass device. It consists of a microfabricated "U"-shape floating platinum electrode placed across the separation channel. The legs of the U function respectively as working and counter electrode. The required potential difference for the ECL reaction is generated at the Pt electrode by the electric field available in the separation channel during electrophoretic separation. Initial experiments demonstrate a micellar electrokinetic separation and direct ECL detection of 10(-16) mol of Ru(phen) (10(-6) M) and 4.5 x 10(-16) mol of Ru(bpy) (5 x 10(-6) M). Also, preliminary results show the indirect detection of three amino acids. The high voltage at the location of detection does not interfere with the electrochemistry.


Clinical Nocardia species: Identification, clinical characteristics, and antimicrobial susceptibility in Shandong, China.
Nocardia is a pathogen responsible for a variety of clinical infections. Here, we aimed to investigate the species distribution, clinical manifestations, and antimicrobial susceptibility of Nocardia species over 3 years in two tertiary general hospitals in China. In this retrospective study, a total of 27 Nocardia species were isolated from 27 individuals between January 2017 and December 2019. Nocardia isolates were identified to species level by mass spectrometry and 16S rRNA PCR sequencing. Clinical data were collected from medical records. Antimicrobial susceptibility was determined by the standard Broth microdilution method. The 27 patients with Nocardia infection included 12 males and 15 females with a mean age of 60.11 years. Among 27 Nocardia isolates, 7 species were identified, with the most common species being Nocardia otitidiscaviarum (40.7%). The antimicrobial susceptibility profiles varied between different Nocardia species. Notably, all Nocardia isolates were linezolid susceptible. The majority of Nocardia isolates were collected from a department of respiratory medicine (55.56%) and sputum specimen (44.44%). Pulmonary region was the most involved body site (70.37%) followed by skin (7.4%) and pleural cavity (7.4%). Most patients with Nocardia infection needed combination antibiotic therapy. Two deaths were reported during the treatment period and 24 patients achieved improvement after antibiotic therapy. The clinical manifestations of Nocardia infection and antimicrobial susceptibility profiles varied with diverse Nocardia species. Thus, the accurate identification of these species is crucial for the diagnosis and the selection of antibiotic treatment.


The emerging role of SOX2 in cell proliferation and survival and its crosstalk with oncogenic signaling in lung cancer.
Tumor cells have long been observed to share several biological characteristics with normal stem/progenitor cells; however, the oncogenic mechanisms underlying the lung stem/progenitor cell signaling remain elusive. Here, we report that SOX2, a self-renewal factor in lung stem/progenitor cells, is highly expressed in a subclass of lung cancer cells, the proliferation, survival, and chemoresistance of which are dependent on SOX2 signaling. Overexpression of SOX2 promotes oncogenic phenotypes in lung cancer cells; knockdown of SOX2 attenuated cell proliferation. We observed that SOX2 increased the expression of epidermal growth factor receptor (EGFR), and EGFR activation further upregulated SOX2 levels, forming a positive feedback loop. SOX2 expression promoted chemoresistance, and silencing of SOX2 perturbed mitochondrial function, causing marked apoptosis and autophagy. SOX2 induced BCL2L1, the ectopic expression of which rescued the effects of SOX2 silencing on apoptosis, autophagy, and mitochondrial function. SOX2 promoted tumor formation, along with increased cell proliferation in a xenograft mouse model. SOX2 expression is associated with poor prognosis in lung cancer patients; moreover, SOX2, EGFR, and BCL2L1 expression levels were significantly correlated in lung tumors. Our findings support the emerging role of SOX2 in cell proliferation and survival by eliciting oncogenic EGFR and BCL2L1 signaling with potential applications as a prognosis marker and a therapeutic target in lung cancer.


Optical properties of metal-dielectric-metal microcavities in the THz frequency range.
We present an experimental and theoretical study of the optical properties of metal-dielectric-metal structures with patterned top metallic surfaces, in the THz frequency range. When the thickness of the dielectric slab is very small with respect to the wavelength, these structures are able to support strongly localized electromagnetic modes, concentrated in the subwavelength metal-metal regions. We provide a detailed analysis of the physical mechanisms which give rise to these photonic modes. Furthermore, our model quantitatively predicts the resonance positions and their coupling to free space photons. We demonstrate that these structures provide an efficient and controllable way to convert the energy of far field propagating waves into near field energy.


Predictors of Acne Vulgaris among Adolescents in Kuwait.
The prevalence of acne vulgaris (AV) among adolescents in Kuwait has remained mostly unknown. This study aimed to estimate the prevalence of AV and its predictors among adolescents in Kuwait. A cross-sectional study was conducted on 714 participants who were randomly selected from public schools using multistage cluster random sampling. The presence or absence of AV, as well as severity of AV, were assessed using the Global Acne Grading Scale. Data on risk factors and potential confounders were collected from the parents through a self-administered questionnaire, and from the adolescents by face-to-face interview. Weight and height were measured in a standardized manner. Of 714 participants, 320 (44.8%; 95% CI 41.1-48.6%) self-reported AV. By clinical examination, AV was found among 479 (67.1%; 95% CI 63.5-70.5%) participants. Factors that were found to be significantly associated with AV in multivariable analysis were age, gender, nationality, and BMI categories. Females compared to males (adjusted prevalence ratio [APR] 1.16 [95% CI 1.05-1.25]; p = 0.006) and overweight or obese adolescents compared to normal weight (APR 1.15 [95% CI 1.02-1.26] and 1.14 [95% CI 1.02-1.23], respectively; p = 0.026) were more likely to have AV. None of the food items studied showed an association with AV in multivariable analysis. The prevalence of self-reported AV seems to be lower than that reported in other settings, while the prevalence of AV by clinical examination resembles that reported in many other countries. BMI was the only modifiable risk factor associated with AV in multivariable analysis, and none of the food items studied was related to the presence or absence of AV in multivariable analysis.


Cerebral blood flow response to flavanol-rich cocoa in healthy elderly humans.
Cerebral ischemia is a common, morbid condition accompanied by cognitive decline. Recent reports on the vascular health benefits of flavanol-containing foods signify a promising approach to the treatment of cerebral ischemia. Our study was designed to investigate the effects of flavanol-rich cocoa (FRC) consumption on cerebral blood flow in older healthy volunteers. We used transcranial Doppler (TCD) ultrasound to measure mean blood flow velocity (MFV) in the middle cerebral artery (MCA) in thirty-four healthy elderly volunteers (72 +/- 6 years) in response to the regular intake of FRC or flavanol-poor cocoa (FPC). In response to two weeks of FRC intake, MFV increased by 8% +/- 4% at one week (p = 0.01) and 10% +/- 4% (p = 0.04) at two weeks. In response to one week of cocoa, significantly more subjects in the FRC as compared with the FPC group had an increase in their MFV (p < 0.05). In summary, we show that dietary intake of FRC is associated with a significant increase in cerebral blood flow velocity in the MCA as measured by TCD. Our data suggest a promising role for regular cocoa flavanol's consumption in the treatment of cerebrovascular ischemic syndromes, including dementias and stroke.


Updated protocol for the examination of specimens removed from patients with colorectal carcinoma. A basis for checklists.
The Cancer Committee of the College of American Pathologists has updated and expanded their protocol for the pathologic examination and reporting of specimens from patients with colorectal carcinoma, which was originally developed in 1989. The updated protocol incorporates all basic pathology data of diagnostic and prognostic significance appropriate for the treatment of patients with colorectal carcinoma. The purpose of the protocol is to serve as a basis for the development of checklists, as an outline for full narrative reporting, as a basis for research protocols, or as a guide for other types of synoptic or reporting formats. The protocol is stratified to accommodate the surgical procedures usually employed for colorectal carcinomas, including incisional endoscopic biopsy, polypectomy, local excision (transanal disc excision), and colon/colorectal resection (eg, segmental resection, total colectomy, abdominoperoneal resection). Explanatory notes detailing specific procedures and rationales for documentation of specific pathologic data are included in the protocol. The protocol uses the recently revised TNM staging system for colorectal carcinoma defined by the American Joint Committee on Cancer and the International Union Against Cancer.


A Closer Look at the Bivariate Association between Ambient Air Pollution and Allergic Diseases: The Role of Spatial Analysis.
Although previous ecological studies investigating the association between air pollution and allergic diseases accounted for temporal or seasonal relationships, few studies address spatial non-stationarity or autocorrelation explicitly. Our objective was to examine bivariate correlation between outdoor air pollutants and the prevalence of allergic diseases, highlighting the limitation of a non-spatial correlation measure, and suggesting an alternative to address spatial autocorrelation. The 5-year prevalence data (2011⁻2015) of allergic rhinitis, atopic dermatitis, and asthma were integrated with the measures of four major air pollutants (SO₂, NO₂, CO, and PM10) for each of the 423 sub-districts of Seoul. Lee's L statistics, which captures how much bivariate associations are spatially clustered, was calculated and compared with Pearson's correlation coefficient for each pair of the air pollutants and allergic diseases. A series of maps showing spatiotemporal patterns of allergic diseases at the sub-district level reveals a substantial degree of spatial heterogeneity. A high spatial autocorrelation was observed for all pollutants and diseases, leading to significant dissimilarities between the two bivariate association measures. The local L statistics identifies the areas where a specific air pollutant is considered to be contributing to a type of allergic disease. This study suggests that a bivariate correlation measure between air pollutants and allergic diseases should capture spatially-clustered phenomenon of the association, and detect the local instability in their relationships. It highlights the role of spatial analysis in investigating the contribution of the local-level spatiotemporal dynamics of air pollution to trends and the distribution of allergic diseases.


Living in living cities.
This article presents an overview of current and potential applications of living technology to some urban problems. Living technology can be described as technology that exhibits the core features of living systems. These features can be useful to solve dynamic problems. In particular, urban problems concerning mobility, logistics, telecommunications, governance, safety, sustainability, and society and culture are presented, and solutions involving living technology are reviewed. A methodology for developing living technology is mentioned, and supraoptimal public transportation systems are used as a case study to illustrate the benefits of urban living technology. Finally, the usefulness of describing cities as living systems is discussed.


Multisensory integration of speech signals: the relationship between space and time.
Integrating audiovisual cues for simple events is affected when sources are separated in space and time. By contrast, audiovisual perception of speech appears resilient when either spatial or temporal disparities exist. We investigated whether speech perception is sensitive to the combination of spatial and temporal inconsistencies. Participants heard the bisyllable /aba/ while seeing a face produce the incongruent bisyllable /ava/. We tested the level of visual influence over auditory perception when the sound was asynchronous with respect to facial motion (from -360 to +360 ms) and emanated from five locations equidistant to the participant. Although an interaction was observed, it was not related to participants' perception of synchrony, nor did it indicate a linear relationship between the effect of spatial and temporal discrepancies. We conclude that either the complexity of the signal or the nature of the task reduces reliance on spatial and temporal contiguity for audiovisual speech perception.


Best bang for your buck: GPU nodes for GROMACS biomolecular simulations.
The molecular dynamics simulation package GROMACS runs efficiently on a wide variety of hardware from commodity workstations to high performance computing clusters. Hardware features are well-exploited with a combination of single instruction multiple data, multithreading, and message passing interface (MPI)-based single program multiple data/multiple program multiple data parallelism while graphics processing units (GPUs) can be used as accelerators to compute interactions off-loaded from the CPU. Here, we evaluate which hardware produces trajectories with GROMACS 4.6 or 5.0 in the most economical way. We have assembled and benchmarked compute nodes with various CPU/GPU combinations to identify optimal compositions in terms of raw trajectory production rate, performance-to-price ratio, energy efficiency, and several other criteria. Although hardware prices are naturally subject to trends and fluctuations, general tendencies are clearly visible. Adding any type of GPU significantly boosts a node's simulation performance. For inexpensive consumer-class GPUs this improvement equally reflects in the performance-to-price ratio. Although memory issues in consumer-class GPUs could pass unnoticed as these cards do not support error checking and correction memory, unreliable GPUs can be sorted out with memory checking tools. Apart from the obvious determinants for cost-efficiency like hardware expenses and raw performance, the energy consumption of a node is a major cost factor. Over the typical hardware lifetime until replacement of a few years, the costs for electrical power and cooling can become larger than the costs of the hardware itself. Taking that into account, nodes with a well-balanced ratio of CPU and consumer-class GPU resources produce the maximum amount of GROMACS trajectory over their lifetime.


Nonneutralizing antibodies binding to the surface glycoprotein of lymphocytic choriomeningitis virus reduce early virus spread.
The biological relevance of nonneutralizing antibodies elicited early after infection with noncytopathic persistence-prone viruses is unclear. We demonstrate that cytotoxic T lymphocyte-deficient TgH(KL25) mice, which are transgenic for the heavy chain of the lymphocytic choriomeningitis virus (LCMV)-neutralizing monoclonal antibody KL25, mount a focused neutralizing antibody response following LCMV infection, and that this results in the emergence of neutralization escape virus variants. Further investigation revealed that some of the escape variants that arose early after infection could still bind to the selecting antibody. In contrast, no antibody binding could be detected for late isolates, indicating that binding, but nonneutralizing, antibodies exerted a selective pressure on the virus. Infection of naive TgH(KL25) mice with distinct escape viruses differing in their antibody-binding properties revealed that nonneutralizing antibodies accelerated clearance of antibody-binding virus variants in a partly complement-dependent manner. Virus variants that did not bind antibodies were not affected. We therefore conclude that nonneutralizing antibodies binding to the same antigenic site as neutralizing antibodies are biologically relevant by limiting early viral spread.


What happened when Scottish women were given advance supplies of emergency contraception? A survey and qualitative study of women's views and experiences.
The Lothian Emergency Contraception Project in Scotland was a radical intervention in which women aged 16-29 were given 5 packs of emergency contraception (EC) to keep at home. We use survey and qualitative interview data to describe how women used the project packs and their views of advance supplies. The women's accounts suggest that concerns that eased access to emergency contraception will lead to repeated use and risky sex appear to be largely unfounded. Women were pleased to be offered the packs, which were reported as having practical advantages and also sparing them the difficulty of negotiating a sometimes awkward consultation. Respondents explained how they used their packs of EC and in their accounts used justifications, repetition and distancing to emphasise that they would not take risks with contraception or sexually transmitted infections. We interpret the data in the light of the observation that EC has an anomalous role in contraception and the work of applied linguists Candlin and Lucas who have demonstrated the difficulties inherent in the family planning consultation.


Safety and efficacy of rivastigmine in adolescents with Down syndrome: long-term follow-up.
Following the completion of a 20-week, open-label study of the safety and efficacy of liquid rivastigmine for adolescents with Down syndrome, 5 of the 10 adolescents in the clinical trial continued long-term rivastigmine therapy and 5 did not. After an average period of 38 months, all 10 subjects returned for a follow-up assessment to determine the safety and efficacy of long-term rivastigmine use. Rivastigmine was well tolerated and overall health appeared to be unaffected by long-term rivastigmine use. Performance change on cognitive and language measures administered at the termination of the open-label clinical trial was compared between the two groups. No between-group difference in median performance change across the long-term period was found, suggesting that the long-term use of rivastigmine does not improve cognitive and language performance. However, two subjects demonstrated remarkable improvement in adaptive function over the long-term period. Both subjects had received long-term rivastigmine therapy. The discussion addresses the challenge of assessing cognitive change in clinical trials using adolescents with Down syndrome as subjects and the use of group versus individual data to evaluate the relevance of medication effects.


Effects of hypertension and hypercholesterolemia on vasodilatation in the rabbit.
Vasodilator substances act either directly on vascular smooth muscle (e.g., adenosine) or indirectly (e.g., acetylcholine) on endothelial cells that respond by releasing an unknown powerful, short-lived relaxing factor. To determine whether chronic hypertension or hypercholesterolemia or both would alter the release of the endothelium-derived relaxing factor, experiments were performed in hypertensive rabbits (5-week cellophane wrap perinephritis; mean blood pressure, 134.7 mm Hg) and normotensive rabbits (mean blood pressure, 80 mm Hg) with a Doppler flow transducer and perivascular balloon implanted on the lower abdominal aorta. Rabbits were fed either 1% cholesterol or control diet for 4 weeks before the experiment. On the day of the experiment, resting hindlimb vascular resistance was greatest in hypertensive rabbits fed 1% cholesterol diet, followed (in descending order) by hypertensive rabbits, normotensive rabbits fed 1% cholesterol diet, and normotensive rabbits. Pharmacological autonomic reflex blockade was induced, and steady state intravenous infusion curves to acetylcholine, serotonin, and adenosine were constructed. Sensitivity (location of effective dose, 50%) to the three vasodilator agents was altered less than twofold from the values in normotensive rabbits for any treatment group. The maximum vasodilator response to acetylcholine, but not to adenosine or serotonin, infusion was reduced significantly in the treated rabbits compared with that in normal rabbits. Reactive hyperemic responses to 5 to 80 seconds of ischemia were not significantly different among the treatment groups. These results indicate that hypertension with or without hypercholesterolemia does not greatly alter the responsiveness of the hindlimb resistance vasculature to these three vasodilator agents or to ischemia.


Orbitally induced oscillations in the East Antarctic ice sheet at the Oligocene/Miocene boundary.
Between 34 and 15 million years (Myr) ago, when planetary temperatures were 3-4 degrees C warmer than at present and atmospheric CO2 concentrations were twice as high as today, the Antarctic ice sheets may have been unstable. Oxygen isotope records from deep-sea sediment cores suggest that during this time fluctuations in global temperatures and high-latitude continental ice volumes were influenced by orbital cycles. But it has hitherto not been possible to calibrate the inferred changes in ice volume with direct evidence for oscillations of the Antarctic ice sheets. Here we present sediment data from shallow marine cores in the western Ross Sea that exhibit well dated cyclic variations, and which link the extent of the East Antarctic ice sheet directly to orbital cycles during the Oligocene/Miocene transition (24.1-23.7 Myr ago). Three rapidly deposited glacimarine sequences are constrained to a period of less than 450 kyr by our age model, suggesting that orbital influences at the frequencies of obliquity (40 kyr) and eccentricity (125 kyr) controlled the oscillations of the ice margin at that time. An erosional hiatus covering 250 kyr provides direct evidence for a major episode of global cooling and ice-sheet expansion about 23.7 Myr ago, which had previously been inferred from oxygen isotope data (Mi1 event).


Cytotoxic Immunity in Peripheral Nerve Injury and Pain.
Cytotoxicity and consequent cell death pathways are a critical component of the immune response to infection, disease or injury. While numerous examples of inflammation causing neuronal sensitization and pain have been described, there is a growing appreciation of the role of cytotoxic immunity in response to painful nerve injury. In this review we highlight the functions of cytotoxic immune effector cells, focusing in particular on natural killer (NK) cells, and describe the consequent action of these cells in the injured nerve as well as other chronic pain conditions and peripheral neuropathies. We describe how targeted delivery of cytotoxic factors via the immune synapse operates alongside Wallerian degeneration to allow local axon degeneration in the absence of cell death and is well-placed to support the restoration of homeostasis within the nerve. We also summarize the evidence for the expression of endogenous ligands and receptors on injured nerve targets and infiltrating immune cells that facilitate direct neuro-immune interactions, as well as modulation of the surrounding immune milieu. A number of chronic pain and peripheral neuropathies appear comorbid with a loss of function of cellular cytotoxicity suggesting such mechanisms may actually help to resolve neuropathic pain. Thus while the immune response to peripheral nerve injury is a major driver of maladaptive pain, it is simultaneously capable of directing resolution of injury in part through the pathways of cellular cytotoxicity. Our growing knowledge in tuning immune function away from inflammation toward recovery from nerve injury therefore holds promise for interventions aimed at preventing the transition from acute to chronic pain.


The effects of MDCO-2010, a serine protease inhibitor, on activated clotting time in blood obtained from volunteers and cardiac surgical patients.
The activated clotting time (ACT) is widely used for monitoring heparin anticoagulation during cardiac surgery. Celite-based ACT values are prolonged when aprotinin is administered. MDCO-2010, a novel serine protease inhibitor, is currently being evaluated as a possible alternative to aprotinin. Therefore, we evaluated the in vitro effects of this novel agent on ACT values using 3 different point-of-care instruments with kaolin or celite as an activator. The study was performed in 2 parts. In the first part, blood samples were obtained from 15 healthy volunteers. Samples were pipetted into small Eppendorf tubes and 2 concentrations of the MDCO-2010 (100 and 500 nM, final concentration) alone or with heparin (1.2 or 2.4 U/mL) were added. ACTs were measured using Helena (celite), Hemochron (kaolin), and Medtronic (kaolin) devices. In the second part of the study, blood samples were obtained intraoperatively, at 5 time points, from 15 patients undergoing cardiopulmonary bypass. MDCO-2010 at a final concentration of 100 or 500 nM was added and ACT testing was performed as before. Additional coagulation tests included prothrombin time, activated partial thromboplastin time, fibrinogen, antithrombin, prothrombin, and anti-Xa levels. Addition of MDCO-2010 concentration-dependently prolonged ACTs in volunteers' and patients' blood samples regardless of the ACT activator or device used. In volunteer samples (no heparin) and in patient samples (baseline and intensive care unit) percent changes in ACTs due to MDCO-2010 were on average 3.1 ± 1.8 times higher (95% confidence interval 2.6-3.6; P < 0.001) for the celite-based Helena device compared with either Hemochron or Medtronic devices. MDCO-2010 causes less ACT prolongation with kaolin than with celite activation.


Emotional and behavioural problems amongst Afghan unaccompanied asylum-seeking children: results from a large-scale cross-sectional study.
Unaccompanied asylum-seeking children (UASC) are considered at high risk for mental health problems, yet few studies focus on single ethnic populations. This study presents results from the largest Afghan UASC mental health survey in the U.K. Specifically, the study aims to estimate the prevalence of emotional and behavioural problems and to investigate the associations of these problems with demographic variables, cumulative traumatic events, and care and migration variables. A census sample of 222 Afghan UASC was interviewed using validated self-report screening measures. Emotional and behavioural problems were screened using the Hopkins Symptoms Checklist 37A (HSCL-37A). Pre-migration stressful life events were screened using the Stressful Life Events Questionnaire. Administrative data on care and asylum were provided by the local authority social services and the UK Border Agency. Approximately one-third (31.4%) scored above cut-offs for emotional and behavioural problems, 34.6% for anxiety and 23.4% for depression. Ordinary least squares regression indicated a significant dose-response relationship between total pre-migration traumatic events and distress as well as between increased time in the country and greater behavioural problems. Compound traumatic events in the pre-migration stages of forced migration have a deleterious association with UASC well-being. Increased time in country suggests a possible peer effect for these children. Consistent with other studies on refugee children, it should be stressed that the majority of UASC scored below suggested cut-offs, thus displaying a marked resilience despite the experience of adverse events.


Effect of host diet on production of organic acids and methane by cockroach gut bacteria.
The effect of high-fiber diets on microbial populations and processes in cockroach guts was investigated by feeding American cockroaches (Periplaneta americana) milled cereal leaves, milled corn cob, or commercial bran-type breakfast cereal in place of the commonly used laboratory diet of dog chow. The activities and numbers of specific gut bacteria varied significantly with the insect's diet and developmental stage. Acetate and lactate were the principal organic acids present in the gut fluid of adult cockroaches and occurred at concentrations of up to 17 and 8 mM, respectively. These acids were most abundant in the gut fluid of dog chow-fed insects, and the greatest amounts were generally found in the foregut and midgut regions. Foreguts of dog chow-fed cockroaches contained an abundant population of lactic acid bacteria that formed acetate and lactate from endogenous hexoses present in the foregut. When adult cockroaches were fed dog chow amended with antibacterial drugs, (i) the concentrations of acetate, lactate, and total hexoses in gut fluid decreased significantly, (ii) the numbers of lactic acid bacteria in the foregut also decreased significantly, and (iii) the production of acetate and lactate by foregut homogenates was suppressed. It was estimated that acetate and lactate produced by bacteria in the foregut of dog chow-fed adult P. americana could support up to 14% of the insect's respiratory requirement if taken up and used by the animal. When insects were fed high-fiber diets of bran cereal, cereal leaves, or corn cob, bacterial production of acetate and lactate in the foregut diminished.(ABSTRACT TRUNCATED AT 250 WORDS)


Understanding transmission and control of the pork tapeworm with CystiAgent: a spatially explicit agent-based model.
The pork tapeworm, Taenia solium, is a serious public health problem in rural low-resource areas of Latin America, Africa and Asia, where the associated conditions of nuerocysticercosis (NCC) and porcine cysticercosis cause substantial health and economic harms. An accurate and validated transmission model for T. solium would serve as an important new tool for control and elimination, as it would allow for comparison of available intervention strategies, and prioritization of the most effective strategies for control and elimination efforts. We developed a spatially-explicit agent-based model (ABM) for T. solium ("CystiAgent") that differs from prior T. solium models by including a spatial framework and behavioral parameters such as pig roaming, open human defecation, and human travel. In this article, we introduce the structure and function of the model, describe the data sources used to parameterize the model, and apply sensitivity analyses (Latin hypercube sampling-partial rank correlation coefficient (LHS-PRCC)) to evaluate model parameters. LHS-PRCC analysis of CystiAgent found that the parameters with the greatest impact on model uncertainty were the roaming range of pigs, the infectious duration of human taeniasis, use of latrines, and the set of "tuning" parameters defining the probabilities of infection in humans and pigs given exposure to T. solium. CystiAgent is a novel ABM that has the ability to model spatial and behavioral features of T. solium transmission not available in other models. There is a small set of impactful model parameters that contribute uncertainty to the model and may impact the accuracy of model projections. Field and laboratory studies to better understand these key components of transmission may help reduce uncertainty, while current applications of CystiAgent may consider calibration of these parameters to improve model performance. These results will ultimately allow for improved interpretation of model validation results, and usage of the model to compare available control and elimination strategies for T. solium.


ω-3 polyunsaturated fatty acids ameliorate type 1 diabetes and autoimmunity.
Despite the benefit of insulin, blockade of autoimmune attack and regeneration of pancreatic islets are ultimate goals for the complete cure of type 1 diabetes (T1D). Long-term consumption of ω-3 polyunsaturated fatty acids (PUFAs) is known to suppress inflammatory processes, making these fatty acids candidates for the prevention and amelioration of autoimmune diseases. Here, we explored the preventative and therapeutic effects of ω-3 PUFAs on T1D. In NOD mice, dietary intervention with ω-3 PUFAs sharply reduced the incidence of T1D, modulated the differentiation of Th cells and Tregs, and decreased the levels of IFN-γ, IL-17, IL-6, and TNF-α. ω-3 PUFAs exerted similar effects on the differentiation of CD4+ T cells isolated from human peripheral blood mononuclear cells. The regulation of CD4+ T cell differentiation was mediated at least in part through ω-3 PUFA eicosanoid derivatives and by mTOR complex 1 (mTORC1) inhibition. Importantly, therapeutic intervention in NOD mice through nutritional supplementation or lentivirus-mediated expression of an ω-3 fatty acid desaturase, mfat-1, normalized blood glucose and insulin levels for at least 182 days, blocked the development of autoimmunity, prevented lymphocyte infiltration into regenerated islets, and sharply elevated the expression of the β cell markers pancreatic and duodenal homeobox 1 (Pdx1) and paired box 4 (Pax4). The findings suggest that ω-3 PUFAs could potentially serve as a therapeutic modality for T1D.


Why own an exotic pet?
Even though people have owned a wide variety of companion animals since times of old, the modern craze for increasingly exotic and little-known species raises a number of questions, including some of an ethical nature. While trade in exotic animals is certainly profitable for these who practise it, it poses great risks of varying types: ecological risks, threats to biodiversity conservation and health risks. Several introduced animal populations have gone on to establish a line in their new host country. We are just starting to measure the adverse impact this has had, in some cases on a very large scale. The veterinary profession doubtless has a major role to play in endeavouring to reform this trade in living creatures that unfortunately results in many losses.


Substrate elasticity regulates skeletal muscle stem cell self-renewal in culture.
Stem cells that naturally reside in adult tissues, such as muscle stem cells (MuSCs), exhibit robust regenerative capacity in vivo that is rapidly lost in culture. Using a bioengineered substrate to recapitulate key biophysical and biochemical niche features in conjunction with a highly automated single-cell tracking algorithm, we show that substrate elasticity is a potent regulator of MuSC fate in culture. Unlike MuSCs on rigid plastic dishes (approximately 10(6) kilopascals), MuSCs cultured on soft hydrogel substrates that mimic the elasticity of muscle (12 kilopascals) self-renew in vitro and contribute extensively to muscle regeneration when subsequently transplanted into mice and assayed histologically and quantitatively by noninvasive bioluminescence imaging. Our studies provide novel evidence that by recapitulating physiological tissue rigidity, propagation of adult muscle stem cells is possible, enabling future cell-based therapies for muscle-wasting diseases.


Directional migration of mesenchymal stem cells under an SDF-1α gradient on a microfluidic device.
Homing of peripheral stem cells is regulated by one of the most representative homing factors, stromal cell-derived factor 1 alpha (SDF-1α), which specifically binds to the plasma membrane receptor CXCR4 of mesenchymal stem cells (MSCs) in order to initiate the signaling pathways that lead to directional migration and homing of stem cells. This complex homing process and directional migration of stem cells have been mimicked on a microfluidic device that is capable of generating a chemokine gradient within the collagen matrix and embedding endothelial cell (EC) monolayers to mimic blood vessels. On the microfluidic device, stem cells showed directional migration toward the higher concentration of SDF-1α, whereas treatment with the CXCR4 antagonist AMD3100 caused loss of directionality of stem cells. Furthermore, inhibition of stem cell's main migratory signaling pathways, Rho-ROCK and Rac pathways, caused blockage of actomyosin and lamellipodia formation, decreasing the migration distance but maintaining directionality. Stem cell homing regulated by SDF-1α caused directional migration of stem cells, while the migratory ability was affected by the activation of migration-related signaling pathways.


Genomic analysis of bone marrow failure and myelodysplastic syndromes reveals phenotypic and diagnostic complexity.
Accurate and timely diagnosis of inherited bone marrow failure and inherited myelodysplastic syndromes is essential to guide clinical management. Distinguishing inherited from acquired bone marrow failure/myelodysplastic syndrome poses a significant clinical challenge. At present, diagnostic genetic testing for inherited bone marrow failure/myelodysplastic syndrome is performed gene-by-gene, guided by clinical and laboratory evaluation. We hypothesized that standard clinically-directed genetic testing misses patients with cryptic or atypical presentations of inherited bone marrow failure/myelodysplastic syndrome. In order to screen simultaneously for mutations of all classes in bone marrow failure/myelodysplastic syndrome genes, we developed and validated a panel of 85 genes for targeted capture and multiplexed massively parallel sequencing. In patients with clinical diagnoses of Fanconi anemia, genomic analysis resolved subtype assignment, including those of patients with inconclusive complementation test results. Eight out of 71 patients with idiopathic bone marrow failure or myelodysplastic syndrome were found to harbor damaging germline mutations in GATA2, RUNX1, DKC1, or LIG4. All 8 of these patients lacked classical clinical stigmata or laboratory findings of these syndromes and only 4 had a family history suggestive of inherited disease. These results reflect the extensive genetic heterogeneity and phenotypic complexity of bone marrow failure/myelodysplastic syndrome phenotypes. This study supports the integration of broad unbiased genetic screening into the diagnostic workup of children and young adults with bone marrow failure and myelodysplastic syndromes.


Airway compromise due to wound hematoma following anterior cervical spine surgery.
One of the most serious adverse events associated with anterior cervical spine surgery is wound hematoma resulting in airway compromise. The reported incidence of this postoperative complication has varied from 0.2% to 1.9%. Obstruction of the airway secondary to bleeding presents a challenging clinical scenario given the rapidity of onset, distorted anatomy of the upper respiratory tract, urgent need to act and potential for catastrophic consequences. This high-risk, life-threatening clinical scenario requires specialized knowledge and a well-designed treatment protocol to achieve a positive outcome. In this review, we report a case of airway compromise secondary to wound hematoma following anterior cervical discectomy and fusion, followed by a review of relevant literature, anatomy, etiologic factors and diagnostic considerations. We also propose guidelines for the prevention and management of postoperative airway obstruction due to wound hematoma.


Immunology of COVID-19: Mechanisms, clinical outcome, diagnostics, and perspectives-A report of the European Academy of Allergy and Clinical Immunology (EAACI).
With the worldwide spread of the novel severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) resulting in declaration of a pandemic by the World Health Organization (WHO) on March 11, 2020, the SARS-CoV-2-induced coronavirus disease-19 (COVID-19) has become one of the main challenges of our times. The high infection rate and the severe disease course led to major safety and social restriction measures worldwide. There is an urgent need of unbiased expert knowledge guiding the development of efficient treatment and prevention strategies. This report summarizes current immunological data on mechanisms associated with the SARS-CoV-2 infection and COVID-19 development and progression to the most severe forms. We characterize the differences between adequate innate and adaptive immune response in mild disease and the deep immune dysfunction in the severe multiorgan disease. The similarities of the human immune response to SARS-CoV-2 and the SARS-CoV and MERS-CoV are underlined. We also summarize known and potential SARS-CoV-2 receptors on epithelial barriers, immune cells, endothelium and clinically involved organs such as lung, gut, kidney, cardiovascular, and neuronal system. Finally, we discuss the known and potential mechanisms underlying the involvement of comorbidities, gender, and age in development of COVID-19. Consequently, we highlight the knowledge gaps and urgent research requirements to provide a quick roadmap for ongoing and needed COVID-19 studies.


Matrix Metalloproteinases in Diabetic Kidney Disease.
Around the world diabetic kidney disease (DKD) is the main cause of chronic kidney disease (CKD), which is characterized by mesangial expansion, glomerulosclerosis, tubular atrophy, and interstitial fibrosis. The hallmark of the pathogenesis of DKD is an increased extracellular matrix (ECM) accumulation causing thickening of the glomerular and tubular basement membranes, mesangial expansion, sclerosis, and tubulointerstitial fibrosis. The matrix metalloproteases (MMPs) family are composed of zinc-dependent enzymes involved in the degradation and hydrolysis of ECM components. Several MMPs are expressed in the kidney; nephron compartments, vasculature and connective tissue. Given their important role in DKD, several studies have been performed in patients with DKD proposing that the measurement of their activity in serum or in urine may become in the future markers of early DKD. Studies from diabetic nephropathy experimental models suggest that a balance between MMPs levels and their inhibitors is needed to maintain renal homeostasis. This review focuses in the importance of the MMPs within the kidney and their modifications at the circulation, kidney and urine in patients with DKD. We also cover the most important studies performed in experimental models of diabetes in terms of MMPs levels, renal expression and its down-regulation effect.


Early experiences matter: a review of the effects of prenatal environment on offspring characteristics in poultry.
Early life experiences can be important in determining offspring phenotypes and may influence interaction with the environment and hence health, welfare, and productivity. The prenatal environment of poultry can be divided into the pre-lay environment and the egg storage/incubation environment, both of which can affect offspring outcomes. The ability to separate maternal and egg/incubation effects makes birds well suited to this type of research. There are many factors, including feeding and nutrition, environmental conditions, husbandry practices, housing system, social environment, infectious environment, and maternal health status, that can influence both the health and performance and behavior and cognition of the offspring. There are some aspects of the environments that can be changed to produce beneficial effects in the offspring, like addition of certain additives to feed or short changes in incubation temperatures, while other aspects should be avoided to reduce negative effects, such as unpredictable feeding and lighting regimens. Measures of offspring characteristics may prove to be a useful method of assessing parent stock welfare if known stressors result in predictable offspring outcomes. This has the advantage of assessing the parent environment without interfering with the animals and possibly affecting their responses and could lead to improved welfare for the animals.


Chronic stress decreases the expression of sympathetic markers in the pineal gland and increases plasma melatonin concentration in rats.
Chronic stress affects brain areas involved in learning and emotional responses. Although most studies have concentrated on the effect of stress on limbic-related brain structures, in this study we investigated whether chronic stress might induce impairments in diencephalic structures associated with limbic components of the stress response. Specifically, we analyzed the effect of chronic immobilization stress on the expression of sympathetic markers in the rat epithalamic pineal gland by immunohistochemistry and western blot, whereas the plasma melatonin concentration was determined by radioimmunoassay. We found that chronic stress decreased the expression of three sympathetic markers in the pineal gland, tyrosine hydroxylase, the p75 neurotrophin receptor and alpha-tubulin, while the same treatment did not affect the expression of the non-specific sympathetic markers Erk1 and Erk2, and glyceraldehyde-3-phosphate dehydrogenase. Furthermore, these results were correlated with a significant increase in plasma melatonin concentration in stressed rats when compared with control animals. Our findings indicate that stress may impair pineal sympathetic inputs, leading to an abnormal melatonin release that may contribute to environmental maladaptation. In addition, we propose that the pineal gland is a target of glucocorticoid damage during stress.


Factors contributing to decreased protein stability when aspartic acid residues are in beta-sheet regions.
Asp residues are significantly under represented in beta-sheet regions of proteins, especially in the middle of beta-strands, as found by a number of studies using statistical, modeling, or experimental methods. To further understand the reasons for this under representation of Asp, we prepared and analyzed mutants of a beta-domain. Two Gln residues of the immunoglobulin light-chain variable domain (V(L)) of protein Len were replaced with Asp, and then the effects of these changes on protein stability and protein structure were studied. The replacement of Q38D, located at the end of a beta-strand, and that of Q89D, located in the middle of a beta-strand, reduced the stability of the parent immunoglobulin V(L) domain by 2.0 kcal/mol and 5.3 kcal/mol, respectively. Because the Q89D mutant of the wild-type V(L)-Len domain was too unstable to be expressed as a soluble protein, we prepared the Q89D mutant in a triple mutant background, V(L)-Len M4L/Y27dD/T94H, which was 4.2 kcal/mol more stable than the wild-type V(L)-Len domain. The structures of mutants V(L)-Len Q38D and V(L)-Len Q89D/M4L/Y27dD/T94H were determined by X-ray diffraction at 1.6 A resolution. We found no major perturbances in the structures of these Q-->D mutant proteins relative to structures of the parent proteins. The observed stability changes have to be accounted for by cumulative effects of the following several factors: (1) by changes in main-chain dihedral angles and in side-chain rotomers, (2) by close contacts between some atoms, and, most significantly, (3) by the unfavorable electrostatic interactions between the Asp side chain and the carbonyls of the main chain. We show that the Asn side chain, which is of similar size but neutral, is less destabilizing. The detrimental effect of Asp within a beta-sheet of an immunoglobulin-type domain can have very serious consequences. A somatic mutation of a beta-strand residue to Asp could prevent the expression of the domain both in vitro and in vivo, or it could contribute to the pathogenic potential of the protein in vivo.


Acute porphyrias: clinical spectrum of hospitalized patients.
To determine characteristics, clinical features and triggers of acute porphyria in hospitalized patients presenting to a tertiary care center in Pakistan. Case series. The Aga Khan University Hospital, Karachi, from 1988 to 2003. Case records of 26 patients hospitalized with diagnosis were identified through computerized hospital patients' data. The diagnosis of acute porphyria was based on pertinent clinical features and laboratory investigations after exclusion of other alternative diagnosis and patients previously diagnosed as porphyric. The data was analyzed through SPSS software version 11.0. Twelve patients (46.2%) were males. Mean age was 21 years. Most common manifestation were gastrointestinal (n=22; 88.5%) followed by neurological symptoms (n=14; 54%). Neurological manifestations included seizures (n=9; 34.6%) and neuropathy (n=6; 23%). One patient presented with depression and insomnia. Family history was positive in (n=8; 30.8%). Eighteen (69%) had history of previous attacks at their presentation to the hospital. Most common precipitating factor was 'eating outside' (n=18; 69%). Porphyrias are uncommon and cryptic group of diseases. This study shows a slightly different gender distribution, earlier onset of symptoms, higher number of neuropsychiatric symptoms (especially seizures), more distal neuropathies and different precipitant in the studied subset of patients than described previously in the western studies.


Giant-cell arteritis in the southern United States. An 11-year retrospective study from the Texas Gulf Coast.
Giant-cell arteritis (GCA) and polymyalgia rheumatica are systemic disorders that reportedly affect primarily white women older than age 50 years. We conducted an 11-year chart review to determine the relative occurrence and pattern of demographic involvement of GCA in the Gulf Coast region of the United States. Of 101,239 computer-coded entries for individual patients aged 40 years or older, 60 charts listed GCA as a differential diagnosis. Twenty-seven patients had temporal GCA; 21 temporal artery biopsy specimens were identified. Two patients had associated systemic GCA (one with aortitis). A striking finding was that 13 of the 27 patients were black women (about 50% of the entire study population). The group with GCA and polymyalgia rheumatica (17 patients) had a significantly higher mean erythrocyte sedimentation rate than the group with "pure" GCA. Jaw claudication and blindness were rare. We concluded that temporal GCA seems relatively uncommon in the Gulf Coast region and in the southern United States as a whole. Furthermore, GCA seems rare in Hispanics (only one patient identified). Nonetheless, this is the first report to document a proportionally high occurrence of GCA in black patients in this part of the country.


Discovery of new inhibitors of Schistosoma mansoni PNP by pharmacophore-based virtual screening.
Schistosomiasis is considered the second most important tropical parasitic disease, with severe socioeconomic consequences for millions of people worldwide. Schistosoma mansoni , one of the causative agents of human schistosomiasis, is unable to synthesize purine nucleotides de novo, which makes the enzymes of the purine salvage pathway important targets for antischistosomal drug development. In the present work, we describe the development of a pharmacophore model for ligands of S. mansoni purine nucleoside phosphorylase (SmPNP) as well as a pharmacophore-based virtual screening approach, which resulted in the identification of three thioxothiazolidinones (1-3) with substantial in vitro inhibitory activity against SmPNP. Synthesis, biochemical evaluation, and structure-activity relationship investigations led to the successful development of a small set of thioxothiazolidinone derivatives harboring a novel chemical scaffold as new competitive inhibitors of SmPNP at the low-micromolar range. Seven compounds were identified with IC(50) values below 100 μM. The most potent inhibitors 7, 10, and 17 with IC(50) of 2, 18, and 38 μM, respectively, could represent new potential lead compounds for further development of the therapy of schistosomiasis.


G-spot anatomy: a new discovery.
The anatomic existence of the G-spot has not been documented yet. To identify the anatomic structure of the G-spot. A stratum-by-stratum vaginal wall dissection on a fresh cadaver. Primary outcome is the identification of the G-spot and the secondary outcome is its measurements and anatomic description of the G-spot. The G-spot has a distinguishable anatomic structure that is located on the dorsal perineal membrane, 16.5 mm from the upper part of the urethral meatus, and creates a 35° angle with the lateral border of the urethra. The lower pole (tail) and the upper pole (head) were located 3 and 15 mm next to the lateral border of the urethra, respectively. Grossly, the G-spot appeared as a well-delineated sac with walls that resembled fibroconnective tissues and resembled erectile tissues. The superior surface of the sac had bluish irregularities visible through the coat. Upon opening the sac's upper coat, blue grape-like anatomic compositions of the G-spot emerged with dimensions of length (L) of 8.1 mm × width (W) of 3.6-1.5 mm × height (H) of 0.4 mm. The G-spot structure had three distinct areas: the proximal part (the head) L 3.4 mm × W 3.6 mm, the middle part L 3.1 mm × W 3.3 mm, and the distal part (tail) L 3.3 mm × W 3.0 mm. From the distal tail, a rope-like structure emerged, which was seen for approximately 1.6 mm and then disappeared into the surrounding tissue. The anatomic existence of the G-spot was documented with potential impact on the practice and clinical research in the field of female sexual function.


Thyroid autoregulation: effect of iodine on glucose transport in cultured thyroid cells.
The nonmetabolizable glucose analogs, [3H]2-deoxy-D-glucose and [3H]O-methyl-D-glucose, were used to determine whether iodide influences glucose transport in porcine cells in primary culture. Incubation with iodide (3 h) decreased basal glucose transport with a half-maximum at NaI 3 X 10(-5) M and maximum at 10(-4) M. Iodide (10(-6) M to 10(-4) M) also abolished the stimulatory effect of TSH (1 mU/ml) on glucose transport. The iodide effect on [3H]2-deoxy-D-glucose transport had the following characteristics: 1) it was abolished 24 h after incubation in iodide-free medium; 2) it was prevented by methimazole (3 mM), and correlated with newly formed organic iodine, 3) and it affected the maximum velocity (Vmax) of glucose transport, reducing it from 25.1 to 14.4 and 12.0 nmol/(min mg protein) at 10(-5) M and 10(-4) M NaI, without affecting the Michaelis-Menten constant (Km) (6mM). Iodide-treated cells had a reduced specific binding of [3H]cytochalasin B (38% and 47% with respect to control cells at 10(-5) M and 10(-4) M NaI). These data suggest that iodide treatment reduces the functional carriers mediating glucose transport in the thyroid.


Building Capacity in Healthcare by Re-examining Clinical Services in Paramedicine.
Emergency departments (ED) continue to be overburdened, leading to crowding and elevated risk of negative clinical outcomes. Extending clinical services to paramedics may support efforts to improve ED burdens by promoting health care access and capacity during times of patient crisis. The objective of this study was to identify the clinical course and most responsible diagnosis of patients transported by paramedic services to local EDs to then evaluate impact of various augmented 9-1-1/paramedic clinical service models on the need for additional ED services. A retrospective cohort and model-simulation based study. We retrieved clinical data from hospital records for a random selection of 3,000 patients who engaged 9-1-1/paramedic services and were transported to a regional ED to identify their clinical course (interventions, diagnostics) disposition and most responsible admitting/discharge diagnosis. We used this data to establish, simulate and test numerous paramedic service models on the need for ED services. A random selection of 3,000 patients was reviewed across 3 hospitals. The majority were female (57.2%) with a mean age of 65 (SD = 21.3). The majority (n = 1954; 65.1%) were discharged directly from ED of which 3.6% (n = 108) received no intervention or diagnostic, 20.4% (n = 611) received only a diagnostic, 4.8% (n = 143) received only an intervention and 36.4% (n = 1092) received both an intervention and diagnostic. The proportion of nonadmitted patients rose to 82.2% and 77.2% when considering lower priority patients and age greater than 65, respectively. Patient types were identified based on frequency and association with discharge directly from ED. Twelve simulated augmented paramedic clinical service models are reported with estimated gains in the number of patients who may no longer require ED services ranging from 7.5% (n = 146) to 35.4% (n = 691). This study suggests a reduction in need for ED services may be achieved through innovative models of paramedic services at the time of crisis. Identifying and confirming patient types/events to target and clinical services to include in the model requires ongoing investigation. Future research will be needed to evaluate the accuracy and impact of the models presented.


Open-labeled study of unilateral autologous bone-marrow-derived mesenchymal stem cell transplantation in Parkinson's disease.
Parkinson's disease (PD) is a progressive neurodegenerative disease for which stem cell research has created hope in the last few years. Seven PD patients aged 22 to 62 years with a mean duration of disease 14.7+/-7.56 years were enrolled to participate in the prospective, uncontrolled, pilot study of single-dose, unilateral transplantation of autologous bone-marrow-derived mesenchymal stem cells (BM-MSCs). The BM-MSCs were transplanted into the sublateral ventricular zone by stereotaxic surgery. Patients were followed up for a period that ranged from 10 to 36 months. The mean baseline "off" score was 65+/-22.06, and the mean baseline "on" score was 50.6+/-15.85. Three of 7 patients have shown a steady improvement in their "off"/"on" Unified Parkinson's Disease Rating Scale (UPDRS). The mean "off" score at their last follow-up was 43.3 with an improvement of 22.9% from the baseline. The mean "on" score at their last follow-up was 31.7, with an improvement of 38%. Hoehn and Yahr (H&Y) and Schwab and England (S&E) scores showed similar improvements from 2.7 and 2.5 in H&Y and 14% improvement in S&E scores, respectively. A subjective improvement was found in symptoms like facial expression, gait, and freezing episodes; 2 patients have significantly reduced the dosages of PD medicine. These results indicate that our protocol seems to be safe, and no serious adverse events occurred after stem-cell transplantation in PD patients. The number of patients recruited and the uncontrolled nature of the trial did not permit demonstration of effectiveness of the treatment involved. However, the results encourage future trials with more patients to demonstrate efficacy.


Carbonic anhydrase I (CA1) is involved in the process of bone formation and is susceptible to ankylosing spondylitis.
Ankylosing spondylitis (AS) is characterized by abnormal bone formation in the spine and the sacroiliac joints. In vitro assays demonstrate that carbonic anhydrase I (CA1) promotes calcium precipitation. This study investigated the function of CA1 for bio-mineralization and determined if common polymorphisms in the CA1 gene might contribute to AS risk. Calcification was induced in Saos-2 cells, a human osteosarcoma cell line, with ascorbic acid and β-glycerophosphate. Calcification was determined by Alizarin Red-S (AR-S) staining. Expressions of CA1, alkaline phosphatase (ALP), bone sialoprotein (BSP), osteocalcin (OCN), osterix (OSX) and runt-related transcription factor-2 (Runx2) were determined by real-time PCR and western blotting. The cells were also treated with acetazolamide, an anti-carbonic anhydrase drug. Genotyping was performed using Illumina VeraCode microarray in a case-control study including 51 AS patients, 267 rheumatoid arthritis (RA) patients and 160 healthy controls. The result was confirmed by Taqman assay, including 258 AS patients, 288 RA patients and 288 healthy controls. Following the induction of calcification, Saos-2 cells produced large amounts of calcium-rich deposits. Increased transcriptions of CA1, ALP, BSP, OCN, OSX and Runx2, essential genes for ossification, were detected in the cultured cells. Following treatmen with acetazolamide, the expression of CA1 obviously declined and mineralized nodule formation was also decreased. Illumina microarray indicates that SNP at rs7841425 also showed significant differences in allelic frequency (P = 0.01396) and genotypic frequency (P = 0.005902) between AS cases and controls. In addition, SNP at rs7827474 showed significant differences in allelic frequency (P = 5.83E-04) and genotypic frequency (P = 0.000186) between RA cases and controls (P values were adjusted to multiple comparisons). The Taqman assay revealed that rs725605 demonstrated statistically significant evidence of allele frequency (P = 0.022307) and gene frequency (P = 0.007731) for association with AS. This SNP did not show significant differences in allelic frequencies and gene frequencies between RA patients and controls. CA1 may play an essential role in bio-mineralization and new bone formation. The gene encoding CA1 is susceptible to AS.


Superresolution laser beam shaping.
The superresolution technique is well known for its ability to compress the central diffractive spot that is smaller than the Airy diffractive spot. In this paper, we extend the superresolution technique for different laser beam shaping. A complete set of superresolution diffractive elements is developed for the flat-top beam shaping, the single-circle beam shaping, and the novel circular Dammann grating. Five phase plates, corresponding to each of its applications, have been made by use of micro-optics technology. Experiments that are presented are in good agreement with the theoretical results. The superresolution technique presented in this paper should be highly interesting for the wide applications of laser beam shaping.


Genetic diversity of influenza A viruses circulating in Bulgaria during the 2018-2019 winter season.
Introduction. Influenza viruses evolve rapidly and change their antigenic characteristics, necessitating biannual updates of flu vaccines.Aim. The aim of this study was to characterize influenza viruses circulating in Bulgaria during the 2018/2019 season and to identify amino acid substitutions in them that might impact vaccine effectiveness.Methodology. Typing/subtyping of influenza viruses were performed using real-time Reverse Transcription-PCR (RT-PCR) and results of phylogenetic and amino acid sequence analyses of influenza strains are presented.Results. A(H1N1)pdm09 (66 %) predominated over A(H3N2) (34 %) viruses, with undetected circulation of B viruses in the 2018/2019 season. All A(H1N1)pdm09 viruses studied fell into the recently designated 6B.1A subclade with over 50 % falling in four subgroups: 6B.1A2, 6B.1A5, 6B.1A6 and 6B.1A7. Analysed A(H3N2) viruses belonged to subclades 3C.2a1b and 3C.2a2. Amino acid sequence analysis of 36 A(H1N1)pdm09 isolates revealed the presence of six-ten substitutions in haemagglutinin (HA), compared to the A/Michigan/45/2015 vaccine virus, three of which occurred in antigenic sites Sa and Cb, together with four-nine changes at positions in neuraminidase (NA), and a number of substitutions in internal proteins. HA1 D222N substitution, associated with increased virulence, was identified in two A(H1N1)pdm09 viruses. Despite the presence of several amino acid substitutions, A(H1N1)pdm09 viruses remained antigenically similar to the vaccine virus. The 28 A(H3N2) viruses characterized carried substitutions in HA, including some in antigenic sites A, B, C and E, in NA and internal protein sequences.Conclusion. The results of this study showed the genetic diversity of circulating influenza viruses and the need for continuous antigenic and molecular surveillance.


A subcortical maternal complex essential for preimplantation mouse embryogenesis.
We have identified a subcortical maternal complex (SCMC) that assembles during oocyte growth and is essential for zygotes to progress beyond the first embryonic cell divisions. At least four maternally encoded proteins contribute to this MDa complex: FLOPED, MATER, and TLE6 interact with each other while Filia binds independently to MATER. Although the transcripts encoding these proteins are degraded during meiotic maturation and ovulation, the SCMC proteins persist in the early embryo. The SCMC, located in the subcortex of eggs, is excluded from regions of cell-cell contact in the cleavage-stage embryo and segregates to the outer cells of the morulae and blastocyst. Floped(tm/tm) and/or Mater(tm/tm) eggs lack the SCMC but can be fertilized. However, these embryos do not progress beyond cleavage stage development and female mice are sterile. The proteins are conserved in humans, and similar maternal effect mutations may result in recurrent embryonic loss.


Effectiveness and cost-effectiveness of prognostic markers in prostate cancer.
This paper demonstrates how economic modelling can be used to derive estimates of the cost-effectiveness of prognostic markers in the management of clinically localised and moderately graded prostate cancer. The model uses a Markov process and is populated using published evidence and local data. The robustness of the results has been tested using sensitivity analysis. Three treatment policies of 'monitoring' (observation), radical prostatectomy, or a selection-based management policy using DNA-ploidy as an experimental marker, have been evaluated. Modelling indicates that a policy of managing these tumours utilising experimental markers has an estimated cost per quality-adjusted life year (QALY) of pound 12 068. Sensitivity analysis shows the results to be relatively sensitive to quality-of-life variables. If novel and experimental markers can achieve specificity in excess of 80%, then a policy of radical surgery for those identified as being at high risk and conservative treatment for the remainder would be both better for patients and cost-effective. The analysis suggests that a radical prostatectomy treatment policy for the moderately graded tumours (Gleason grades -7) modelled in this paper may be inferior to a conservative approach in the absence of reliable prognostic markers, being both more costly and yielding fewer QALYs.


Family, Community, and Health System Considerations for Reducing the Burden of Pediatric Sickle Cell Disease in Uganda Through Newborn Screening.
Sickle cell disease (SCD) is associated with high mortality for children under 5 years of age in sub-Saharan Africa. Newborn sickle screening program and enhanced capacity for SCD treatment are under development to reduce disease burden in Uganda and elsewhere in the region. Based on an international stakeholder meeting and a family-directed conference on SCD in Kampala in 2015, and interviews with parents, multinational experts, and other key informants, we describe health care, community, and family perspectives in support of these initiatives. Key stakeholder meetings, discussions, and interviews were held to understand perspectives of public health and multinational leadership, patients and families, as well as national progress, resource needs, medical and social barriers to program success, and resources leveraged from HIV/AIDS. Partnering with program leadership, professionals, patients and families, multinational stakeholders, and leveraging resources from existing programs are needed for building successful programs in Uganda and elsewhere in sub-Saharan Africa.


Primary vascular bone tumors in the spine: a challenge for pathologists and spine oncology surgeons.
Primary vascular bone tumors of the spine represent a challenge for oncology surgeons, due to the need of planning a surgical strategy appropriate to the tumor behavior. But these tumors represent a challenge also for pathologists, as immunohistochemical and molecular analyses have recently refined the terminology. A cohort of 81 cases was retrospectively reviewed, targeting the evolution of diagnoses and the treatment-related outcome. Sixty-six cases (including one case originally diagnosed as chordoma) were diagnosed before 2011, and 15 were diagnosed and treated in the period 2011-2017. Fully documented outcome studies are available for 46 patients whose immunohistochemical and molecular analyses were available. The follow-up ranges from 6 months of the early patient death to 300 months of the longest disease-free survival. The outcome was related not only to the treatment performed, but also to the evolution of diagnoses. The term Hemangioendothelioma that defined a benign aggressive (Enneking stage 3) tumor is now obsolete and replaced by the diagnosis of Epithelioid Hemangioma for benign aggressive tumor and Epithelioid Hemangioendothelioma for low-grade malignant tumor. En bloc resection was appropriate for local control, but 2 out of 7 cases of epithelioid Hemangioendotheliomas died due to the spread of the disease None of the Hemangiomas recurred after intralesional excision with or without radiotherapy, or after vertebroplasty. Both cases of Angiosarcoma had a fast and lethal evolution. Surgeons must be aware of the evolution of terminology to decide the most appropriate treatment options. These slides can be retrieved from Electronic Supplementary Material.


Regulation of clonal growth by anti-T-cell receptor antibody-directed lysis.
CD4 and CD8 T-cell clones were generated using the Mx9 monoclonal antibody (mAb), which recognizes the V beta 8 T-cell receptor (TcR) gene family. The interaction of these clones with the Mx9 antibody was analysed and all were found to be specifically stimulated to proliferate by plastic-adherent Mx9. In the presence of Mx9 or its F(ab')2 fragment, CD8+ Mx9+ clones were capable of specifically lysing the CD4+ Mx9+ T-cell clones. No lysis was seen of Mx9- T cells, or in the absence of the antibody. Conversely CD4+ Mx9+ T cells did not have lytic function. These results indicate that cross-linking of T cells via their antigen-specific receptors may initiate a unidirectional killing. Unlike previously reported lytic systems involving anti-TcR antibodies (e.g. anti-CD3), these results suggest that this mechanism may have an important physiological role in immune regulation. Anti-idiotypic antibodies have been shown to recognize T-cell receptors. These may exert profound immunosuppressive effects by inducing the lysis of the helper cells or B cells.


Resveratrol promotes recovery of immune function of immunosuppressive mice by activating JNK/NF-κB pathway in splenic lymphocytes.
Resveratrol, a natural compound found in over 70 plants, is known to possess immunoregulatory effects and anti-inflammatory activity. It has been shown that resveratrol has regulatory effects on different signaling pathways in different diseases. However, few reports have evaluated the effects of resveratrol on reinforcing immunity recovery via activating nuclear factor-κB (NF-κB) pathway and Jun N-terminal kinases (JNK) pathway. The present study aimed to assess immune-enhancing activity and underlying mechanism of resveratrol in immunosuppressive mice. Previously, we reported that resveratrol could promote mouse spleen lymphocyte functions to recover the immune system effectively. In the present study, we show that resveratrol could upregulate the expressions of NF-κB, IκB kinase, JNK, and c-jun in splenic lymphocytes of immunosuppressive mice. Taken together, our results indicate that resveratrol could promote recovery of immunologic function in immunosuppressive mice by activating JNK/NF-κB pathway.


Impact of asian ethnicity on colorectal cancer screening: a population-based analysis.
Although research shows that African Americans and Hispanics frequently receive less colorectal cancer screening (CRCS) than whites, few studies have focused on CRCS among Asians. The aims of this study were to compare CRCS between Asians and whites and to evaluate for clinical predictors of CRCS. From the 2007 California Health Interview Survey, we identified all Asian and white respondents who were eligible for CRCS. Logistic regression was performed to evaluate for differences in CRCS. We used stratified and interaction analyses to examine whether associations between race and CRCS were modified by insurance status, birthplace, or language skills, while controlling for other confounders. Baseline characteristics were similar between Asians and whites. Only 58% of Asians and 66% of whites reported undergoing up-to-date CRCS (P < 0.01). In multivariate analyses, visiting a physician more than 5 times produced the highest odds of being up-to-date with screening. When compared with whites, Asians had decreased odds of being up-to-date with screening. Stratified analyses showed that this disparity existed mainly in the insured, but not in the uninsured, and it was not modified by place of birth or English language proficiency. Despite its ability to reduce mortality, CRCS is suboptimal in our US population-based cohort of Asians when compared with whites. A contributing factor to this problem for the Chinese and Koreans may be a lack of awareness regarding CRCS, whereas the source of the problem in the Vietnamese seems to be related to healthcare access.


Cross-talk between aryl hydrocarbon receptor and the inflammatory response: a role for nuclear factor-κB.
The aryl hydrocarbon receptor (AhR) is involved in the regulation of immune responses, T-cell differentiation, and immunity. Here, we show that inflammatory stimuli such as LPS induce the expression of AhR in human dendritic cells (DC) associated with an AhR-dependent increase of CYP1A1 (cytochrome P4501A1). In vivo data confirmed the elevated expression of AhR by LPS and the LPS-enhanced 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-mediated induction of CYP1A1 in thymus of B6 mice. Inhibition of nuclear factor-κB (NF-κB) repressed both normal and LPS-enhanced, TCDD-inducible, AhR-dependent gene expression and canonical pathway control of RelA-regulated AhR-responsive gene expression. LPS-mediated induction of AhR was NF-κB-dependent, as shown in mouse embryonic fibroblasts (MEFs) derived from Rel null mice. AhR expression and TCDD-mediated induction of CYP1A1 was significantly reduced in RelA-deficient MEF compared with wild type MEF cells and ectopic expression of RelA restored the expression of AhR and induction of CYP1A1 in MEF RelA null cells. Promoter analysis of the human AhR gene identified three putative NF-κB-binding elements upstream of the transcription start site. Mutation analysis of the AhR promoter identified one NF-κB site as responsible for mediating the induction of AhR expression by LPS and electrophoretic shift assays demonstrated that this NF-κB motif is recognized by the RelA/p50 heterodimer. Our results show for the first time that NF-κB RelA is a critical component regulating the expression of AhR and the induction of AhR-dependent gene expression in immune cells illustrating the interaction of AhR and NF-κB signaling.


Chronic neuropsychiatric sequelae of SARS-CoV-2: Protocol and methods from the Alzheimer's Association Global Consortium.
Coronavirus disease 2019 (COVID-19) has caused >3.5 million deaths worldwide and affected >160 million people. At least twice as many have been infected but remained asymptomatic or minimally symptomatic. COVID-19 includes central nervous system manifestations mediated by inflammation and cerebrovascular, anoxic, and/or viral neurotoxicity mechanisms. More than one third of patients with COVID-19 develop neurologic problems during the acute phase of the illness, including loss of sense of smell or taste, seizures, and stroke. Damage or functional changes to the brain may result in chronic sequelae. The risk of incident cognitive and neuropsychiatric complications appears independent from the severity of the original pulmonary illness. It behooves the scientific and medical community to attempt to understand the molecular and/or systemic factors linking COVID-19 to neurologic illness, both short and long term. This article describes what is known so far in terms of links among COVID-19, the brain, neurological symptoms, and Alzheimer's disease (AD) and related dementias. We focus on risk factors and possible molecular, inflammatory, and viral mechanisms underlying neurological injury. We also provide a comprehensive description of the Alzheimer's Association Consortium on Chronic Neuropsychiatric Sequelae of SARS-CoV-2 infection (CNS SC2) harmonized methodology to address these questions using a worldwide network of researchers and institutions. Successful harmonization of designs and methods was achieved through a consensus process initially fragmented by specific interest groups (epidemiology, clinical assessments, cognitive evaluation, biomarkers, and neuroimaging). Conclusions from subcommittees were presented to the whole group and discussed extensively. Presently data collection is ongoing at 19 sites in 12 countries representing Asia, Africa, the Americas, and Europe. The Alzheimer's Association Global Consortium harmonized methodology is proposed as a model to study long-term neurocognitive sequelae of SARS-CoV-2 infection. The following review describes what is known so far in terms of molecular and epidemiological links among COVID-19, the brain, neurological symptoms, and AD and related dementias (ADRD)The primary objective of this large-scale collaboration is to clarify the pathogenesis of ADRD and to advance our understanding of the impact of a neurotropic virus on the long-term risk of cognitive decline and other CNS sequelae. No available evidence supports the notion that cognitive impairment after SARS-CoV-2 infection is a form of dementia (ADRD or otherwise). The longitudinal methodologies espoused by the consortium are intended to provide data to answer this question as clearly as possible controlling for possible confounders. Our specific hypothesis is that SARS-CoV-2 triggers ADRD-like pathology following the extended olfactory cortical network (EOCN) in older individuals with specific genetic susceptibility.The proposed harmonization strategies and flexible study designs offer the possibility to include large samples of under-represented racial and ethnic groups, creating a rich set of harmonized cohorts for future studies of the pathophysiology, determinants, long-term consequences, and trends in cognitive aging, ADRD, and vascular disease.We provide a framework for current and future studies to be carried out within the Consortium. and offers a "green paper" to the research community with a very broad, global base of support, on tools suitable for low- and middle-income countries aimed to compare and combine future longitudinal data on the topic.The Consortium proposes a combination of design and statistical methods as a means of approaching causal inference of the COVID-19 neuropsychiatric sequelae. We expect that deep phenotyping of neuropsychiatric sequelae may provide a series of candidate syndromes with phenomenological and biological characterization that can be further explored. By generating high-quality harmonized data across sites we aim to capture both descriptive and, where possible, causal associations.


Spinal Tuberculosis - Directly Observed Treatment and Short Course or Daily Anti Tubercular Therapy -Are We Over Treating?
Prospective randomised control trial. To compare the effectiveness of Directly Observed Treatment and Short Course (DOTS) and Daily Anti Tubercular Therapy (ATT) in spinal tuberculosis with no neurological deficit. Spinal tuberculosis is rampant in India with a major cause of morbidity and mortality. There is a lot of conflict regarding management with anti tubercular drugs, the regimen to be followed and the duration the drugs needed for complete cure. This prospective study was conducted during the period of 2006 to 2009. Thirty cases of spinal tuberculosis were randomly divided equally into two groups of fifteen each and treated with DOTS and Daily ATT and compared at the end of follow up on clinical, radiological and Erythrocyte Sedimentation Rate values (ESR). Pain score on Visual Analogue Scale (VAS) exhibits that mean pain score was 5.93+_1.54 at start and 0.64+_1.01 at the end of follow up with 89.39% change with DOTS therapy whereas mean pain score was 7.08 +_1.61at start of therapy and 0.69+_0.95 at end of follow up with 91.73% change in patients treated with Daily ATT with p value >0.05 (not significant). The radiological recovery in patients with DOTS and Daily ATT have similar results after 1.5 years with p value > 0.05 showing that the radiological recovery by both treatment modalities are comparable. Mean change in ESR scores in DOTS therapy patients and patients on daily ATT were 74.57+_9.34% and 75.69+_9.38% change with p value >0.05 which was not significant. DOTS therapy is an effective means of management of spinal tuberculosis cases with no neurological involvement, however its efficacy in patients with neurological deficit is further to be evaluated.


Implementation and impact of an audit and feedback antimicrobial stewardship intervention in the orthopaedics department of a tertiary-care hospital: a controlled interrupted time series study.
A prospective audit and feedback antimicrobial stewardship intervention conducted in the Orthopaedics Department of a university hospital in Portugal was evaluated by comparing an interrupted time series in the intervention group with a non-intervention (control) group. Monthly antibiotic use (except cefazolin) was measured as the World Health Organization's Anatomical Therapeutic Chemical defined daily doses (ATC-DDD) from January 2012 to September 2016, excluding the 6-month phase of intervention implementation starting on 1 January 2015. Compared with the control group, the intervention group had a monthly decrease in the use of fluoroquinolones by 2.3 DDD/1000 patient-days [95% confidence interval (CI) -3.97 to -0.63]. An increase in the use of penicillins by 103.3 DDD/1000 patient-days (95% CI 47.42 to 159.10) was associated with intervention implementation, followed by a decrease during the intervention period (slope = -5.2, 95% CI -8.56 to -1.82). In the challenging scenario of treatment of osteoarticular and prosthetic joint infections, an audit and feedback intervention reduced antibiotic exposure and spectrum.


FRET-enhanced photostability allows improved single-molecule tracking of proteins and protein complexes in live mammalian cells.
A major challenge in single-molecule imaging is tracking the dynamics of proteins or complexes for long periods of time in the dense environments found in living cells. Here, we introduce the concept of using FRET to enhance the photophysical properties of photo-modulatable (PM) fluorophores commonly used in such studies. By developing novel single-molecule FRET pairs, consisting of a PM donor fluorophore (either mEos3.2 or PA-JF549) next to a photostable acceptor dye JF646, we demonstrate that FRET competes with normal photobleaching kinetic pathways to increase the photostability of both donor fluorophores. This effect was further enhanced using a triplet-state quencher. Our approach allows us to significantly improve single-molecule tracking of chromatin-binding proteins in live mammalian cells. In addition, it provides a novel way to track the localization and dynamics of protein complexes by labeling one protein with the PM donor and its interaction partner with the acceptor dye.


Randomized, double-blind, placebo-controlled study of interferon-γ 1b in Friedreich Ataxia.
In vitro, in vivo, and open-label studies suggest that interferon gamma (IFN-γ 1b) may improve clinical features in Friedreich Ataxia through an increase in frataxin levels. The present study evaluates the efficacy and safety of IFN-γ 1b in the treatment of Friedreich Ataxia through a double-blind, multicenter, placebo-controlled trial. Ninety-two subjects with FRDA between 10 and 25 years of age were enrolled. Subjects received either IFN-γ 1b or placebo for 6 months. The primary outcome measure was the modified Friedreich Ataxia Rating Scale (mFARS). No difference was noted between the groups after 6 months of treatment in the mFARS or secondary outcome measures. No change was noted in buccal cell or whole blood frataxin levels. However, during an open-label extension period, subjects had a more stable course than expected based on natural history data. This study provides no direct evidence for a beneficial effect of IFN-γ1b in FRDA. The modest stabilization compared to natural history data leaves open the possibility that longer studies may demonstrate benefit.


Reducing youth screen time: qualitative metasynthesis of findings on barriers and facilitators.
An integrated perspective on the relevant qualitative findings on the experience of screen time in youth can inform the development of hypotheses to be tested in future research and can guide the development of interventions to decrease sedentary behavior. The purpose of this qualitative metasynthesis was to explore parent, youth, and educational professionals' perceptions of barriers to, and facilitators of, reducing youth screen time. Qualitative metasynthesis techniques were used to analyze and synthesize 15 qualitative studies of screen time among youth (11-18 years) meeting inclusion criteria. The phrases, quotes, and/or author interpretations (i.e., theme or subtheme) were recorded in a data display matrix to facilitate article comparisons. Codes were collapsed into 23 categories of similar conceptual meaning and 3 overarching themes were derived using thematic analysis procedures. Study sample sizes ranged from 6 to 270 participants from 6 countries. Data collection methods included focus groups (n = 6), interviews (n = 4), focus group and interviews (n = 4), and naturalistic observation (n = 1) with youth and/or parents. Data analysis techniques included thematic analysis (n = 9), content analysis (n = 3), grounded theory (n = 1), observation (n = 1), and interpretive phenomenological analysis (n = 1). Three thematic categories were identified: (a) youth's norms-screen time is an integral part of daily life, and facilitates opportunities for entertainment, social interaction, and escapism; (b) family dynamics and parental roles-parents are conflicted and send mixed messages about the appropriate uses and amounts of screen time; and, (c) resources and environment-engagement in screen time is dependent on school, community, neighborhood, and home environmental contexts. Screen time is an established norm in many youth cultures, presenting barriers to behavior change. Parents recognize the importance of reducing youth screen time, but model and promote engagement themselves. For youth and parents, mutually agreed rules, limits, and parental monitoring of screen time were perceived as likely to be effective.


Characterization of 1H NMR spectroscopic data and the generation of synthetic validation sets.
Common contemporary practice within the nuclear magnetic resonance (NMR) metabolomics community is to evaluate and validate novel algorithms on empirical data or simplified simulated data. Empirical data captures the complex characteristics of experimental data, but the optimal or most correct analysis is unknown a priori; therefore, researchers are forced to rely on indirect performance metrics, which are of limited value. In order to achieve fair and complete analysis of competing techniques more exacting metrics are required. Thus, metabolomics researchers often evaluate their algorithms on simplified simulated data with a known answer. Unfortunately, the conclusions obtained on simulated data are only of value if the data sets are complex enough for results to generalize to true experimental data. Ideally, synthetic data should be indistinguishable from empirical data, yet retain a known best analysis. We have developed a technique for creating realistic synthetic metabolomics validation sets based on NMR spectroscopic data. The validation sets are developed by characterizing the salient distributions in sets of empirical spectroscopic data. Using this technique, several validation sets are constructed with a variety of characteristics present in 'real' data. A case study is then presented to compare the relative accuracy of several alignment algorithms using the increased precision afforded by these synthetic data sets. These data sets are available for download at http://birg.cs.wright.edu/nmr_synthetic_data_sets.