Inhibition of Shedding of Low-Density Lipoprotein Receptor-Related Protein 1 Reverses Cartilage Matrix Degradation in Osteoarthritis.
The aggrecanase ADAMTS-5 and the collagenase matrix metalloproteinase 13 (MMP-13) are constitutively secreted by chondrocytes in normal cartilage, but rapidly endocytosed via the cell surface endocytic receptor low-density lipoprotein receptor-related protein 1 (LRP-1) and subsequently degraded. This endocytic system is impaired in osteoarthritic (OA) cartilage due to increased ectodomain shedding of LRP-1. The aim of this study was to identify the LRP-1 sheddase(s) in human cartilage and to test whether inhibition of LRP-1 shedding prevents cartilage degradation in OA. Cell-associated LRP-1 and soluble LRP-1 (sLRP-1) released from human cartilage explants and chondrocytes were measured by Western blot analysis. LRP-1 sheddases were identified by proteinase inhibitor profiling and gene silencing with small interfering RNAs. Specific monoclonal antibodies were used to selectively inhibit the sheddases. Degradation of aggrecan and collagen in human OA cartilage was measured by Western blot analysis using an antibody against an aggrecan neoepitope and a hydroxyproline assay, respectively. Shedding of LRP-1 was increased in OA cartilage compared with normal tissue. Shed sLRP-1 bound to ADAMTS-5 and MMP-13 and prevented their endocytosis without interfering with their proteolytic activities. Two membrane-bound metalloproteinases, ADAM-17 and MMP-14, were identified as the LRP-1 sheddases in cartilage. Inhibition of their activities restored the endocytic capacity of chondrocytes and reduced degradation of aggrecan and collagen in OA cartilage. Shedding of LRP-1 is a key link to OA progression. Local inhibition of LRP-1 sheddase activities of ADAM-17 and MMP-14 is a unique way to reverse matrix degradation in OA cartilage and could be effective as a therapeutic approach.


Antibiotics in severely malnourished children: systematic review of efficacy, safety and pharmacokinetics.
To systemically review the evidence in support of World Health Organization guidelines recommending broad-spectrum antibiotics for children with severe acute malnutrition (SAM). CENTRAL, MEDLINE, EMBASE, LILACS, POPLINE, CAB Abstracts and ongoing trials registers were searched. Experts were contacted. Conference proceedings and reference lists were manually searched. All study types, except single case reports, were included. Two randomized controlled trials (RCTs), one before-and-after study and two retrospective reports on clinical efficacy and safety were retrieved, together with 18 pharmacokinetic studies. Trial quality was generally poor and results could not be pooled due to heterogeneity. Oral amoxicillin for 5 days was as effective as intramuscular ceftriaxone for 2 days (1 RCT). For uncomplicated SAM, amoxicillin showed no benefit over placebo (1 retrospective study). The introduction of a standardized regimen using ampicillin and gentamicin significantly reduced mortality in hospitalized children (odds ratio, OR: 4.0; 95% confidence interval, CI: 1.7-9.8; 1 before-and-after study). Oral chloramphenicol was as effective as trimethoprim-sulfamethoxazole in children with pneumonia (1 RCT). Pharmacokinetic data suggest that normal doses of penicillins, cotrimoxazole and gentamicin are safe in malnourished children, while the dose or frequency of chloramphenicol requires adjustment. Existing evidence is not strong enough to further clarify recommendations for antibiotic treatment in children with SAM. Large RCTs are needed to define optimal antibiotic treatment in children with SAM with and without complications. Further research into gentamicin and chloramphenicol toxicity and into the pharmacokinetics of ceftriaxone and ciprofloxacin is also required.


Ovar-DRB1 haplotypes *2001 and *0301 are associated with sheep growth and ewe lifetime prolificacy.
The major histocompatibility complex (MHC) is an organized cluster of tightly linked vertebrate genes with immunological and non-immunological functions. While the important MHC gene DRB1 has been examined in regard to many sheep infectious disease traits, only one study, based on microsatellite markers, has previously examined DRB1 and sheep production traits. Furthermore, to our knowledge no studies have examined DRB1 relationship with lifetime ewe prolificacy traits. Therefore, we analyzed association between the presence of DRB1 SNP haplotypes with internationally recognized standard names and production traits including growth and lifetime prolificacy in 370 Rambouillet, Columbia, and Polypay sheep. The DRB1 *2001 haplotype was associated with increased weaning and mature weights, as well as average daily gain (Šidák P<0.05; corrected for the number of haplotypes tested). Interestingly, the *2001 haplotype also showed a trend toward association with increased total number of lifetime lambs born (Šidák P=0.084) and number of lambs born alive (Šidák P=0.084). In contrast, the DRB1 *0301 haplotype was associated with decreased mature weight (Šidák P=0.01). Since the *2001 haplotype was present in all three breeds, these results suggest there is at least one functional mutation in the region that influences growth and prolificacy traits that may be broadly present across several breeds. Furthermore, combined use of the similar *2001 and *0301 multi-marker haplotypes that nonetheless have opposing directions of production trait associations will enhance mutation discovery in this region. If undesirable alleles for underlying mutations can be identified, selective pressure against one or a small number of undesirable alleles may improve production with limited impact on MHC genetic diversity and infectious disease susceptibility.


Outcomes following conservative treatment of extension fractures in the setting of diffuse idiopathic skeletal hyperostosis: is external orthosis alone a reasonable option?
Extension fractures in the setting of diffuse idiopathic skeletal hyperostosis (DISH) represent highly unstable injuries. As a result, these fractures are most frequently treated with immediate surgical fixation to limit any potential risk of associated neurological injury. Although this represents the standard of care, patients with significant comorbidities, advanced age, or medical instability may not be surgical candidates. In this paper, the authors evaluated a series of patients with extension DISH fractures who were treated with orthosis alone and evaluated their outcomes. A retrospective review from 2015 to 2022 was conducted at a large level 1 trauma center. Patients with extension-type DISH fractures without neurological deficits were identified. All patients were treated conservatively with orthosis alone. Baseline patient characteristics and adverse outcomes are reported. Twenty-seven patients were identified as presenting with extension fractures associated with DISH without neurological deficit. Of these, 22 patients had complete follow-up on final chart review. Of these 22 patients, 21 (95.5%) were treated successfully with external orthosis. One patient (4.5%) who was noncompliant with the brace had an acute spinal cord injury 1 month after presentation, requiring immediate surgical fixation and decompression. No other complications, including skin breakdown or pressure ulcers related to bracing, were reported. Treatment of extension-type DISH fractures may be a reasonable option for patients who are not candidates for safe surgical intervention; however, a risk of neurological injury secondary to delayed instability remains, particularly if patients are noncompliant with the bracing regimen. This risk should be balanced against the high complication rate and potential mortality associated with surgical intervention in this patient population.


Significant Locus and Metabolic Genetic Correlations Revealed in Genome-Wide Association Study of Anorexia Nervosa.
The authors conducted a genome-wide association study of anorexia nervosa and calculated genetic correlations with a series of psychiatric, educational, and metabolic phenotypes. Following uniform quality control and imputation procedures using the 1000 Genomes Project (phase 3) in 12 case-control cohorts comprising 3,495 anorexia nervosa cases and 10,982 controls, the authors performed standard association analysis followed by a meta-analysis across cohorts. Linkage disequilibrium score regression was used to calculate genome-wide common variant heritability (single-nucleotide polymorphism [SNP]-based heritability [h2SNP]), partitioned heritability, and genetic correlations (rg) between anorexia nervosa and 159 other phenotypes. Results were obtained for 10,641,224 SNPs and insertion-deletion variants with minor allele frequencies >1% and imputation quality scores >0.6. The h2SNP of anorexia nervosa was 0.20 (SE=0.02), suggesting that a substantial fraction of the twin-based heritability arises from common genetic variation. The authors identified one genome-wide significant locus on chromosome 12 (rs4622308) in a region harboring a previously reported type 1 diabetes and autoimmune disorder locus. Significant positive genetic correlations were observed between anorexia nervosa and schizophrenia, neuroticism, educational attainment, and high-density lipoprotein cholesterol, and significant negative genetic correlations were observed between anorexia nervosa and body mass index, insulin, glucose, and lipid phenotypes. Anorexia nervosa is a complex heritable phenotype for which this study has uncovered the first genome-wide significant locus. Anorexia nervosa also has large and significant genetic correlations with both psychiatric phenotypes and metabolic traits. The study results encourage a reconceptualization of this frequently lethal disorder as one with both psychiatric and metabolic etiology.


Pancreatic Inflammation Redirects Acinar to β Cell Reprogramming.
Using a transgenic mouse model to express MafA, Pdx1, and Neurog3 (3TF) in a pancreatic acinar cell- and doxycycline-dependent manner, we discovered that the outcome of transcription factor-mediated acinar to β-like cellular reprogramming is dependent on both the magnitude of 3TF expression and on reprogramming-induced inflammation. Overly robust 3TF expression causes acinar cell necrosis, resulting in marked inflammation and acinar-to-ductal metaplasia. Generation of new β-like cells requires limiting reprogramming-induced inflammation, either by reducing 3TF expression or by eliminating macrophages. The new β-like cells were able to reverse streptozotocin-induced diabetes 6 days after inducing 3TF expression but failed to sustain their function after removal of the reprogramming factors.


Accuracy of transvaginal ultrasound for diagnosis of deep endometriosis in the rectosigmoid: systematic review and meta-analysis.
To review the diagnostic accuracy of transvaginal ultrasound (TVS) in the preoperative detection of rectosigmoid endometriosis in patients with clinical suspicion of deep infiltrating endometriosis (DIE), comparing enhanced (E-TVS) and non-enhanced approaches. An extensive search was performed in MEDLINE (PubMed) and EMBASE for studies published between January 1989 and December 2014. The eligibility criterion was use of TVS for preoperative detection of rectosigmoid endometriosis in women with clinical suspicion of DIE, using surgical data as the reference standard. Study quality was assessed using Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines and the Quality Assessment of Diagnostic Accuracy Studies-2 (QUADAS-2) tool. Our extended search identified a total of 801 citations, among which 19 studies (n = 2639) were considered eligible and included in the meta-analysis. Overall pooled sensitivity, specificity, positive likelihood ratio (LR+) and negative likelihood ratio (LR-) of TVS for detecting DIE in the rectosigmoid were 91% (95%CI, 85-94%), 97% (95%CI, 95-98%), 33.0 (95%CI, 18.6-58.6) and 0.10 (95%CI, 0.06-0.16), respectively. Significant heterogeneity was found for sensitivity (I(2) , 90.8%; Cochran Q, 195.2; P < 0.001) and specificity (I(2) , 76.8%; Cochran Q, 77.7; P < 0.001). We did not find statistical differences between non-enhanced TVS and E-TVS (P = 0.304). Overall diagnostic performance of TVS for DIE of the rectosigmoid is good. However, further studies with improved quality in design are needed. Copyright © 2015 ISUOG. Published by John Wiley & Sons Ltd.


Co-administering Melatonin With an Estradiol-Progesterone Menopausal Hormone Therapy Represses Mammary Cancer Development in a Mouse Model of HER2-Positive Breast Cancer.
Melatonin has numerous anti-cancer properties reported to influence cancer initiation, promotion, and metastasis. With the need for effective hormone therapies (HT) to treat menopausal symptoms without increasing breast cancer risk, co-administration of nocturnal melatonin with a natural, low-dose HT was evaluated in mice that develop primary and metastatic mammary cancer. Individually, melatonin (MEL) and estradiol-progesterone therapy (EPT) did not significantly affect mammary cancer development through age 14 months, but, when combined, the melatonin-estradiol-progesterone therapy (MEPT) significantly repressed tumor formation. This repression was due to effects on tumor incidence, but not latency. These results demonstrate that melatonin and the HT cooperate to decrease the mammary cancer risk. Melatonin and EPT also cooperate to alter the balance of the progesterone receptor (PR) isoforms by significantly increasing PRA protein expression only in MEPT mammary glands. Melatonin significantly suppressed amphiregulin transcripts in MEL and MEPT mammary glands, suggesting that amphiregulin together with the higher PRA:PRB balance and other factors may contribute to reducing cancer development in MEPT mice. Melatonin supplementation influenced mammary morphology by increasing tertiary branching in the mouse mammary glands and differentiation in human mammary epithelial cell cultures. Uterine weight in the luteal phase was elevated after long-term exposure to EPT, but not to MEPT, indicating that melatonin supplementation may reduce estrogen-induced uterine stimulation. Melatonin supplementation significantly decreased the incidence of grossly-detected lung metastases in MEL mice, suggesting that melatonin delays the formation of metastatic lesions and/or decreases aggressiveness in this model of HER2+ breast cancer. Mammary tumor development was similar in EPT and MEPT mice until age 8.6 months, but after 8.6 months, only MEPT continued to suppress cancer development. These data suggest that melatonin supplementation has a negligible effect in young MEPT mice, but is required in older mice to inhibit tumor formation. Since melatonin binding was significantly decreased in older mammary glands, irrespective of treatment, melatonin supplementation may overcome reduced melatonin responsiveness in the aged MEPT mice. Since melatonin levels are known to decline near menopause, nocturnal melatonin supplementation may also be needed in aging women to cooperate with HT to decrease breast cancer risk.


p53-Dependent Senescence in Mesenchymal Stem Cells under Chronic Normoxia Is Potentiated by Low-Dose γ-Irradiation.
Mesenchymal stem cells (MSCs) are a source of adult multipotent cells important in tissue regeneration. Murine MSCs are known to proliferate poorly in vitro under normoxia. The aim of this study is to analyze the interaction of nonphysiological high oxygen and low-dose γ-irradiation onto growth, senescence, and DNA damage. Tri-potent bone marrow-derived MSCs from p53 wildtype and p53-/- mice were cultured under either 21% or 2% O2. Long-term observations revealed a decreasing ability of wildtype mMSCs to proliferate and form colonies under extended culture in normoxia. This was accompanied by increased senescence under normoxia but not associated with telomere shortening. After low-dose γ-irradiation, the normoxic wildtype cells further increased the level of senescence. The number of radiation-induced γH2AX DNA repair foci was higher in mMSCs kept under normoxia but not in p53-/- cells. P53-deficient MSCs additionally showed higher clonogeneity, lower senescence levels, and fewer γH2AX repair foci per cell as compared to their p53 wildtype counterparts irrespective of oxygen levels. These results reveal that oxygen levels together with γ-irradiation and p53 status are interconnected factors modulating growth capacity of BM MSCs in long-term culture. These efforts help to better understand and optimize handling of MSCs prior to their therapeutic use.


Noninvasive carotid strain imaging using angular compounding at large beam steered angles: validation in vessel phantoms.
Stroke and myocardial infarction are initiated by rupturing vulnerable atherosclerotic plaques. With noninvasive ultrasound elastography, these plaques might be detected in carotid arteries. However, since the ultrasound beam is generally not aligned with the radial direction in which the artery pulsates, radial and circumferential strains need to be derived from axial and lateral data. Conventional techniques to perform this conversion have the disadvantage that lateral strain is required. Since the lateral strain has relatively poor accuracy, the quality of the radial and circumferential strains is reduced. In this study, the radial and circumferential strain estimates are improved by combining axial strain data acquired at multiple insonification angles. Adaptive techniques to correct for grating lobe interference and other artifacts that occur when performing beam steering at large angles are introduced. Acquisitions at multiple angles are performed with a beam steered linear array. For each beam steered angle, there are two spatially restricted regions of the circular vessel cross section where the axial strain is closely aligned with the radial strain and two spatially restricted regions (different from the radial strain regions) where the axial strain is closely aligned with the circumferential strain. These segments with high quality strain estimates are compounded to form radial or circumferential strain images. Compound radial and circumferential strain images were constructed for a homogeneous vessel phantom with a concentric lumen subjected to different intraluminal pressures. Comparison of the elastographic signal-to-noise ratio (SNR(e)) and contrast-to-noise ratio (CNR(e)) revealed that compounding increases the image quality considerably compared to images from 0 degrees information only. SNR(e) and CNR(e) increase up to 2.7 and 6.6 dB, respectively. The highest image quality was achieved by projecting axial data, completed with a small segment determined by either principal component analysis or by application of a rotation matrix.


Impaired relaxation despite upregulated calcium-handling protein atrial myocardium from type 2 diabetic patients with preserved ejection fraction.
Diastolic dysfunction is a key factor in the development and pathology of cardiac dysfunction in diabetes, however the exact underlying mechanism remains unknown, especially in humans. We aimed to measure contraction, relaxation, expression of calcium-handling proteins and fibrosis in myocardium of diabetic patients with preserved systolic function. Right atrial appendages from patients with type 2 diabetes mellitus (DM, n = 20) and non-diabetic patients (non-DM, n = 36), all with preserved ejection fraction and undergoing coronary artery bypass grafting (CABG), were collected. From appendages, small cardiac muscles, trabeculae, were isolated to measure basal and β-adrenergic stimulated myocardial function. Expression levels of calcium-handling proteins, sarcoplasmic reticulum Ca2+ ATPase (SERCA2a) and phospholamban (PLB), and of β1-adrenoreceptors were determined in tissue samples by Western blot. Collagen deposition was determined by picro-sirius red staining. In trabeculae from diabetic samples, contractile function was preserved, but relaxation was prolonged (Tau: 74 ± 13 ms vs. 93 ± 16 ms, non-DM vs. DM, p = 0.03). The expression of SERCA2a was increased in diabetic myocardial tissue (0.75 ± 0.09 vs. 1.23 ± 0.15, non-DM vs. DM, p = 0.007), whereas its endogenous inhibitor PLB was reduced (2.21 ± 0.45 vs. 0.42 ± 0.11, non-DM vs. DM, p = 0.01). Collagen deposition was increased in diabetic samples. Moreover, trabeculae from diabetic patients were unresponsive to β-adrenergic stimulation, despite no change in β1-adrenoreceptor expression levels. Human type 2 diabetic atrial myocardium showed increased fibrosis without systolic dysfunction but with impaired relaxation, especially during β-adrenergic challenge. Interestingly, changes in calcium-handling protein expression suggests accelerated active calcium re-uptake, thus improved relaxation, indicating a compensatory calcium-handling mechanism in diabetes in an attempt to maintain diastolic function at rest despite impaired relaxation in the diabetic fibrotic atrial myocardium. Our study addresses important aspects of the underlying mechanisms of diabetes-associated diastolic dysfunction, which is crucial to developing new therapeutic treatments.


[Cognitive communication disorder due to right hemispheric brain injury].
Communication disorders can arise as a consequence of right hemispheric brain injury due to, for example, stroke. These disorders can comprise language comprehension, language production and pragmatic language use. If these problems are not recognised in time, treatment will probably focus on behavioural problems, which would lead to further deterioration of the patient due to an escalation in interaction. We present a patient in whom this situation occured. However, by addressing her underlying communication problem, we were able to improve her quality of life.


An alpaca nanobody inhibits hepatitis C virus entry and cell-to-cell transmission.
Severe liver disease caused by chronic hepatitis C virus is the major indication for liver transplantation. Despite recent advances in antiviral therapy, drug toxicity and unwanted side effects render effective treatment in liver-transplanted patients a challenging task. Virus-specific therapeutic antibodies are generally safe and well-tolerated, but their potential in preventing and treating hepatitis C virus (HCV) infection has not yet been realized due to a variety of issues, not least high production costs and virus variability. Heavy-chain antibodies or nanobodies, produced by camelids, represent an exciting antiviral approach; they can target novel highly conserved epitopes that are inaccessible to normal antibodies, and they are also easy to manipulate and produce. We isolated four distinct nanobodies from a phage-display library generated from an alpaca immunized with HCV E2 glycoprotein. One of them, nanobody D03, recognized a novel epitope overlapping with the epitopes of several broadly neutralizing human monoclonal antibodies. Its crystal structure revealed a long complementarity determining region (CD3) folding over part of the framework that, in conventional antibodies, forms the interface between heavy and light chain. D03 neutralized a panel of retroviral particles pseudotyped with HCV glycoproteins from six genotypes and authentic cell culture-derived particles by interfering with the E2-CD81 interaction. In contrast to some of the most broadly neutralizing human anti-E2 monoclonal antibodies, D03 efficiently inhibited HCV cell-to-cell transmission. This is the first description of a potent and broadly neutralizing HCV-specific nanobody representing a significant advance that will lead to future development of novel entry inhibitors for the treatment and prevention of HCV infection and help our understanding of HCV cell-to-cell transmission.


Stress- and diet-induced fat gain is controlled by NPY in catecholaminergic neurons.
Neuropeptide Y (NPY) and noradrenaline are commonly co-expressed in sympathetic neurons. Both are key regulators of energy homeostasis and critical for stress-coping. However, little is known about the specific function of NPY in the catecholaminergic system in these regulations. Here we show that mice with NPY expression only in the noradrenergic and adrenergic cells of the catecholaminergic system (catNPY) exhibited exacerbated diet-induced obesity, lower body and brown adipose tissue temperatures compared to WT and NPY(-/-) mice under a HFD. Furthermore, chronic stress increased adiposity and serum corticosterone level in WT but not NPY(-/-) mice. Re-introducing NPY specifically to the catecholaminergic system in catNPY mice restored stress responsiveness associated with increased respiratory exchange ratio and decreased liver pACC to tACC ratio. These results demonstrate catecholaminergic NPY signalling is critical in mediating diet- and chronic stress-induced fat gain via effects on diet-induced thermogenesis and stress-induced increases in corticosterone levels and lipogenic capacity.


Transmissible cancers in an evolutionary context.
Cancer is an evolutionary and ecological process in which complex interactions between tumour cells and their environment share many similarities with organismal evolution. Tumour cells with highest adaptive potential have a selective advantage over less fit cells. Naturally occurring transmissible cancers provide an ideal model system for investigating the evolutionary arms race between cancer cells and their surrounding micro-environment and macro-environment. However, the evolutionary landscapes in which contagious cancers reside have not been subjected to comprehensive investigation. Here, we provide a multifocal analysis of transmissible tumour progression and discuss the selection forces that shape it. We demonstrate that transmissible cancers adapt to both their micro-environment and macro-environment, and evolutionary theories applied to organisms are also relevant to these unique diseases. The three naturally occurring transmissible cancers, canine transmissible venereal tumour (CTVT) and Tasmanian devil facial tumour disease (DFTD) and the recently discovered clam leukaemia, exhibit different evolutionary phases: (i) CTVT, the oldest naturally occurring cell line is remarkably stable; (ii) DFTD exhibits the signs of stepwise cancer evolution; and (iii) clam leukaemia shows genetic instability. While all three contagious cancers carry the signature of ongoing and fairly recent adaptations to selective forces, CTVT appears to have reached an evolutionary stalemate with its host, while DFTD and the clam leukaemia appear to be still at a more dynamic phase of their evolution. Parallel investigation of contagious cancer genomes and transcriptomes and of their micro-environment and macro-environment could shed light on the selective forces shaping tumour development at different time points: during the progressive phase and at the endpoint. A greater understanding of transmissible cancers from an evolutionary ecology perspective will provide novel avenues for the prevention and treatment of both contagious and non-communicable cancers.


Core decompression of the distal femur for avascular necrosis of the knee.
From 1974 to 1981, we performed 28 core decompressions of the distal femur for pathologically confirmed avascular necrosis. At a mean follow-up of 54 months (range 20 to 140 months) and using the Ficat stages, all seven cases in stage I and stage II had good results. Of 21 cases in stage III, 11 cases had good results, four had poor results, and six needed total knee replacement. There were no significant orthopaedic complications. The procedure is worthwhile and will be more accurate with new methods of imaging.


Non-genomic effects of xenoestrogen mixtures.
Xenoestrogens (XEs) are chemicals derived from a variety of natural and anthropogenic sources that can interfere with endogenous estrogens by either mimicking or blocking their responses via non-genomic and/or genomic signaling mechanisms. Disruption of estrogens' actions through the less-studied non-genomic pathway can alter such functional end points as cell proliferation, peptide hormone release, catecholamine transport, and apoptosis, among others. Studies of potentially adverse effects due to mixtures and to low doses of endocrine-disrupting chemicals have recently become more feasible, though few so far have included actions via the non-genomic pathway. Physiologic estrogens and XEs evoke non-monotonic dose responses, with different compounds having different patterns of actions dependent on concentration and time, making mixture assessments all the more challenging. In order to understand the spectrum of toxicities and their mechanisms, future work should focus on carefully studying individual and mixture components across a range of concentrations and cellular pathways in a variety of tissue types.


A role for Drosophila Drac1 in neurite outgrowth and synaptogenesis in the giant fiber system.
Recent studies have shown the small GTPases, Rac1, Rho, and CDC42, to have a role in axon guidance. To assess their participation in synapse assembly and function we have expressed various forms of Drac1 in the giant fiber system of Drosophila. Overexpression of wild-type Drac1 in the giant fiber (GF) lead to a disruption in axonal morphology; axons often terminate prematurely in a large swelling in the target area but lack the normal lateral bend where the synapse with the jump motor neuron would normally be found. Electrophysiological assays revealed longer latencies and lowering following frequencies indicating defects in the synapse between the GF and the tergotrochanteral motor neuron (TTMn). Thickened abnormal GF dendrites were also observed in the brain. Overexpression of the dominant-negative form of Drac1, (N17), resulted in axons that produced extra branches in the second thoracic neuromere (T2); however, the synaptic connection to the TTMn was present and functioned normally. Conversely, expression of the constitutively active form, Drac1(V12), resulted in a complete lack of neurite outgrowth and this was also seen with overexpression of Dcdc42(V12). In the absence of a GF, these flies showed no response in the jump (TTM) or flight (DLM) muscles upon brain stimulation. Taken together these results show that the balance of actin polymerization and depolymerization determines local process outgrowth and thereby synapse structure and function.


Radiographic evaluation of factors affecting bearing dislocation in the domed lateral Oxford unicompartmental knee replacement.
The rate of bearing dislocation with the domed lateral Oxford Unicompartmental Knee Replacement (OUKR) in different series varies from 1% to 6% suggesting that dislocation is influenced by surgical technique. The aim of this study was to identify surgical factors associated with dislocation. Aligned post-operative antero-posterior knee radiographs of seven knees that had dislocated and 87 control knees were compared. Component alignment and position and the alignment of the knee were assessed. All bearing dislocations occurred medially over the tibial wall. Knees that dislocated tended to be overcorrected: Compared with those that did not dislocate, they were in 2° less valgus (p=0.019) and the tibial components were positioned 2 mm more proximal (p<0.01). Although the relative position of the centre of the femoral component and the tibial component was the same (p=0.8), in the dislocating group the gap between the edge of the femoral component and the top of the wall in flexion was 3mm greater (p=0.019) suggesting that the components were internally rotated. To minimise the risk of dislocation it is recommended that the knee should not be overstuffed. This is best achieved by selecting the bearing thickness that just tightens the ligaments in full extension, and re-cutting the tibia if necessary. In addition to minimise the gap between the femoral and tibial components through which the bearing dislocates, the femoral component should be implanted in neutral rotation and should not be internally rotated. Level IV.


Detecting Entanglement Structure in Continuous Many-Body Quantum Systems.
A prerequisite for the comprehensive understanding of many-body quantum systems is a characterization in terms of their entanglement structure. The experimental detection of entanglement in spatially extended many-body systems describable by quantum fields still presents a major challenge. We develop a general scheme for certifying entanglement and demonstrate it by revealing entanglement between distinct subsystems of a spinor Bose-Einstein condensate. Our scheme builds on the spatially resolved simultaneous detection of the quantum field in two conjugate observables which allows the experimental confirmation of quantum correlations between local as well as nonlocal partitions of the system. The detection of squeezing in Bogoliubov modes in a multimode setting illustrates its potential to boost the capabilities of quantum simulations to study entanglement in spatially extended many-body systems.


Precision medicine in acute respiratory distress syndrome: workshop report and recommendations for future research.
Acute respiratory distress syndrome (ARDS) is a devastating critical illness that can be triggered by a wide range of insults and remains associated with a high mortality of around 40%. The search for targeted treatment for ARDS has been disappointing, possibly due to the enormous heterogeneity within the syndrome. In this perspective from the European Respiratory Society research seminar on "Precision medicine in ARDS", we will summarise the current evidence for heterogeneity, explore the evidence in favour of precision medicine and provide a roadmap for further research in ARDS. There is evident variation in the presentation of ARDS on three distinct levels: 1) aetiological; 2) physiological and 3) biological, which leads us to the conclusion that there is no typical ARDS. The lack of a common presentation implies that intervention studies in patients with ARDS need to be phenotype aware and apply a precision medicine approach in order to avoid the lack of success in therapeutic trials that we faced in recent decades. Deeper phenotyping and integrative analysis of the sources of variation might result in identification of additional treatable traits that represent specific pathobiological mechanisms, or so-called endotypes.


Performance and image analysis of the aberration-corrected Hitachi HD-2700C STEM.
We report the performance of the first aberration-corrected scanning transmission electron microscope (STEM) manufactured by Hitachi. We describe its unique features and versatile capabilities in atomic-scale characterization and its applications in materials research. We also discuss contrast variation of the STEM images obtained from different annular dark-field (ADF) detectors of the instrument, and the increased complexity in contrast interpretation and quantification due to the increased convergent angles of the electron probe associated with the aberration corrector. We demonstrate that the intensity of atomic columns in an ADF image depends strongly on a variety of imaging parameters, sample thickness, as well as the nuclear charge and the deviation from their periodic position of the atoms we are probing. Image simulations are often required to correctly interpret the atomic structure of an ADF-STEM image.


Evaluating an emergency medical services-initiated nontransport system.
To evaluate both factors predicting nontransport and mortality rates in an emergency medical services system with a nontransport policy. We reviewed data from 1,581 transported and nontransported patients from October 2001 to July 2003. Patients who refused transport against medical advice were excluded. Extracted data included demographics, run characteristics, chief complaint, and clinical impression. Transported and nontransported patients were compared using Mann-Whitney U or chi-square tests. Logistic regression identified factors predictive of nontransport. A Social Security Death Index search determined 30-day mortality. A total of 1,501 runs involving 1,059 patients were included. Median age was 60 years (range, 0-97 years). A total of 427 (40.4%) were male; 107 (10.2%) were nonwhite. Older patients were more likely to be transported (odds ratio, 1.03; confidence interval, 1.02-1.03). Race, frequency of calls, mutual aid, or time of day did not significantly influence probability of transport. Patients with cardiovascular, respiratory, and gastrointestinal complaints were more likely to be transported than those with other conditions (P < 0.005); patients with endocrine, trauma, and miscellaneous complaints were less likely to be transported (P < 0.003). Patients with renal, obstetrics/gynecology, and hema matology/oncology were complaints all transported. Mortality was 4.9% (confidence interval, 3.9%-6.2%) for transported patients and 1.0% for those not transported (confidence interval, 0.2%-3.7%). Age is a determinant when deciding on transporting patients. Patients with complaints with potentially higher acuity were transported most often. Only two nontransported patients died within 30 days, although it is unknown whether initial transport would have changed their mortality. Our data suggest that emergency medical services-initiated nontransport is influenced only by age and chief complaint and may not result in significant mortality.


Finding groups using model-based cluster analysis: heterogeneous emotional self-regulatory processes and heavy alcohol use risk.
Model-based cluster analysis is a new clustering procedure to investigate population heterogeneity utilizing finite mixture multivariate normal densities. It is an inferentially based, statistically principled procedure that allows comparison of nonnested models using the Bayesian information criterion to compare multiple models and identify the optimum number of clusters. The current study clustered 36 young men and women on the basis of their baseline heart rate and heart rate variability (HRV), chronic alcohol use, and reasons for drinking. Two cluster groups were identified and labeled the high alcohol risk and normative groups. Compared to the normative group, individuals in the high alcohol risk group had higher levels of alcohol use and more strongly endorsed disinhibition and suppression reasons for use. The high alcohol risk group showed significant HRV changes in response to positive and negative emotional and appetitive picture cues, compared to neutral cues. In contrast, the normative group showed a significant HRV change only to negative cues. Findings suggest that individuals with autonomic self-regulatory difficulties may be more susceptible to heavy alcohol use and use of alcohol for emotional regulation.


Neural drive increases following resistance training in patients with multiple sclerosis.
The present study tested the hypothesis that lower body progressive resistance training (PRT) increases the neural drive expressed as surface electromyographical (EMG) activity in patients with multiple sclerosis (MS). The study was a randomised controlled trial (RCT) including a 12-week follow up period. Thirty-eight MS patients were randomized to an exercise group (n = 19) or a control group (n = 19). During the intervention period, the exercise group performed a 12-week supervised lower body PRT program, whereas the control group maintained their usual daily activity level. After the 12 week intervention period, the exercise group were encouraged to continue training on their own for a 12-week follow up period, while the control group completed the 12-week supervised PRT program. Surface EMG was recorded from vastus lateralis, rectus femoris and semitendinosus during maximal isometric knee extension and knee flexion, before and after the intervention and at follow up. From the recordings, the area under the rectified, low-pass filtered EMG signal (integrated EMG, iEMG) was calculated. Muscle strength was expressed as the angular impulse (AI) and was measured during the same period as the iEMG. After PRT significant improvements for iEMG of vastus lateralis and rectus femoris during maximal knee extension and for semitendinosus during maximal knee flexion and for AI during both maximal knee extension and flexion were found in the exercise group, when compared to the control group. When compared to the post values, all effects, except for AI during knee flexion, were maintained at follow up in the exercise group. When the control group was exposed to PRT, a similar pattern of improvements were found, albeit not all improvements were significant. In conclusion twelve weeks of intense PRT of the lower extremities improved the neural drive expressed as maximal surface EMG activity in patients with MS, with effects persisting 12 weeks after the intervention. The study was registered at clinicalTrials.gov, Protocol no. NCT00381576.


Identifying the critical success factors in the coverage of low vision services using the classification analysis and regression tree methodology.
To identify the critical success factors (CSF) associated with coverage of low vision services. Data were collected from a survey distributed to Vision 2020 contacts, government, and non-government organizations (NGOs) in 195 countries. The Classification and Regression Tree Analysis (CART) was used to identify the critical success factors of low vision service coverage. Independent variables were sourced from the survey: policies, epidemiology, provision of services, equipment and infrastructure, barriers to services, human resources, and monitoring and evaluation. Socioeconomic and demographic independent variables: health expenditure, population statistics, development status, and human resources in general, were sourced from the World Health Organization (WHO), World Bank, and the United Nations (UN). The findings identified that having >50% of children obtaining devices when prescribed (χ(2) = 44; P < 0.000), multidisciplinary care (χ(2) = 14.54; P = 0.002), >3 rehabilitation workers per 10 million of population (χ(2) = 4.50; P = 0.034), higher percentage of population urbanized (χ(2) = 14.54; P = 0.002), a level of private investment (χ(2) = 14.55; P = 0.015), and being fully funded by government (χ(2) = 6.02; P = 0.014), are critical success factors associated with coverage of low vision services. This study identified the most important predictors for countries with better low vision coverage. The CART is a useful and suitable methodology in survey research and is a novel way to simplify a complex global public health issue in eye care.


Transcription-factor-dependent enhancer transcription defines a gene regulatory network for cardiac rhythm.
The noncoding genome is pervasively transcribed. Noncoding RNAs (ncRNAs) generated from enhancers have been proposed as a general facet of enhancer function and some have been shown to be required for enhancer activity. Here we examine the transcription-factor-(TF)-dependence of ncRNA expression to define enhancers and enhancer-associated ncRNAs that are involved in a TF-dependent regulatory network. TBX5, a cardiac TF, regulates a network of cardiac channel genes to maintain cardiac rhythm. We deep sequenced wildtype and Tbx5-mutant mouse atria, identifying ~2600 novel Tbx5-dependent ncRNAs. Tbx5-dependent ncRNAs were enriched for tissue-specific marks of active enhancers genome-wide. Tbx5-dependent ncRNAs emanated from regions that are enriched for TBX5-binding and that demonstrated Tbx5-dependent enhancer activity. Tbx5-dependent ncRNA transcription provided a quantitative metric of Tbx5-dependent enhancer activity, correlating with target gene expression. We identified RACER, a novel Tbx5-dependent long noncoding RNA (lncRNA) required for the expression of the calcium-handling gene Ryr2. We illustrate that TF-dependent enhancer transcription can illuminate components of TF-dependent gene regulatory networks.


Mig1 localization exhibits biphasic behavior which is controlled by both metabolic and regulatory roles of the sugar kinases.
Glucose, fructose and mannose are the preferred carbon/energy sources for the yeast Saccharomyces cerevisiae. Absence of preferred energy sources activates glucose derepression, which is regulated by the kinase Snf1. Snf1 phosphorylates the transcriptional repressor Mig1, which results in its exit from the nucleus and subsequent derepression of genes. In contrast, Snf1 is inactive when preferred carbon sources are available, which leads to dephosphorylation of Mig1 and its translocation to the nucleus where Mig1 acts as a transcription repressor. Here we revisit the role of the three hexose kinases, Hxk1, Hxk2 and Glk1, in glucose de/repression. We demonstrate that all three sugar kinases initially affect Mig1 nuclear localization upon addition of glucose, fructose and mannose. This initial import of Mig1 into the nucleus was temporary; for continuous nucleocytoplasmic shuttling of Mig1, Hxk2 is required in the presence of glucose and mannose and in the presence of fructose Hxk2 or Hxk1 is required. Our data suggest that Mig1 import following exposure to preferred energy sources is controlled via two different pathways, where (1) the initial import is regulated by signals derived from metabolism and (2) continuous shuttling is regulated by the Hxk2 and Hxk1 proteins. Mig1 nucleocytoplasmic shuttling appears to be important for the maintenance of the repressed state in which Hxk1/2 seems to play an essential role.


HTLV-1 provirus load in peripheral blood lymphocytes of HTLV-1 carriers is diminished by green tea drinking.
Human T-cell lymphotropic virus type 1 (HTLV-1) is causatively associated with adult T-cell leukemia (ATL) and HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). Since a high level of HTLV-1 provirus load in circulating lymphocytes is thought to be a risk for ATL and HAM/TSP, diminution of HTLV-1 provirus load in the circulation may prevent these intractable diseases. Our previous study (Jpn J Cancer Res 2000; 91: 34-40) demonstrated that green tea polyphenols inhibit in vitro growth of ATL cells, as well as HTLV-1-infected T-cells. The present study aimed to investigate the in vivo effect of green tea polyphenols on HTLV-1 provirus load in peripheral blood lymphocytes on HTLV-1 carriers. We recruited 83 asymptomatic HTLV-1 carriers to examine HTLV-1 provirus DNA with or without administration of capsulated green tea extract powder. Thirty-seven subjects were followed up for 5 months by measuring HTLV-1 provirus load after daily intake of 9 capsules of green tea extract powder per day (equivalent to 10 cups of regular green tea), and 46 subjects lived ad libitum without intake of any green tea capsule. The real-time PCR quantification of HTLV-1 DNA revealed a wide range of variation of HTLV-1 provirus load among asymptomatic HTLV-1 carriers (0.2-200.2 copies of HTLV-1 provirus load per 1000 peripheral blood lymphocytes). Daily intake of the capsulated green tea for 5 months significantly diminished the HTLV-1 provirus load as compared with the controls (P = 0.031). These results suggest that green tea drinking suppresses proliferation of HTLV-1-infected lymphocytes in vivo.


Philosophical background of attitudes toward and treatment of invertebrates.
People who interact with or make decisions about invertebrate animals have an attitude toward them, although they may not have consciously worked it out. Three philosophical approaches underlie this attitude. The first is the contractarian, which basically contends that animals are only automata and that we humans need not concern ourselves with their welfare except for our own good, because cruelty and neglect demean us. A second approach is the utilitarian, which focuses on gains versus losses in interactions between animals, including humans. Given the sheer numbers of invertebrates-they constitute 99% of the animals on the planet- this attitude implicitly requires concern for them and consideration in particular of whether they can feel pain. Third is the rights-based approach, which focuses on humans-treatment of animals by calling for an assessment of their quality of life in each human-animal interaction. Here scholars debate to what extent different animals have self-awareness or even consciousness, which may dictate our treatment of them. Regardless of the philosophical approach to invertebrates, information and education about their lives are critical to an understanding of how humans ought to treat them.


FLUORESCENCE POLARIZATION: MEASUREMENT WITH ULTRAVIOLET-POLARIZING FILTERS IN A SPECTROPHOTOFLUOROMETER.
Ultraviolet-polarizing filters with large apertures facilitate the use of spectrophotofluorometers for measuring fluorescence polarization. Such filters (i) allow the use of polarized exciting light even in the ultraviolet region, (ii) make it possible to correct for grating anomalies which influence the observed polarization, and (iii) because of good transmission properties allow the entrance of sufficient light so that small slits and dilute solutions can be used. Under optimum conditions, polarization can be measured with a precision of +/-0.002. Results obtained are in excellent agreement with values cited elsewhere and indicate that standard spectrophotofluorometers can easily be adapted for accurate measurement of polarization.


Microsurgical correction of facial contour deformities in patients with craniofacial malformations: a 15-year experience.
Since their first review of microsurgical correction of facial contour deformities in 19 patients with craniofacial malformations, the authors have treated an additional 74 patients (n = 93). The authors review indications, choices, safety, efficacy, complications, and technical refinements. A treatment algorithm is presented. A retrospective chart review of all patients who underwent microvascular reconstruction of the face and all patients with craniofacial dysmorphology was performed. Between 1989 and 2004, a total of 93 patients with the following diagnoses were identified: craniofacial microsomia (n = 73), Treacher Collins syndrome (n = 8), and severe orbitofacial cleft (n = 12). All patients underwent microsurgical facial reconstruction with a superficial inferior epigastric, groin, or circumflex scapular flap. Flap revisions, complications, and non-free flap related surgery were reviewed. The mean age at microvascular reconstruction was 11 years (range, 4 to 27 years). Flap choices included the following: superficial inferior epigastric (n = 4), groin (n = 3), and circumflex scapular (n = 105). Seventy-six patients underwent unilateral and 17 patients underwent bilateral (one of 17 simultaneous) reconstructions. Postoperative complications included partial flap loss (n = 1), reexploration (n = 1), hematoma (n = 5), and cellulitis (n = 5). All patients had subjective improvement in facial contour, symmetry, skin tone, and color. Most patients underwent additional non-free flap procedures including mandibular distraction and ear reconstruction. Microsurgical flaps have markedly improved the authors' ability to restore craniofacial contour in patients with craniofacial malformations. In selected patients, the authors choose primary midface augmentation with free vascularized tissue to restore form and function. Microsurgical flaps in patients with craniofacial malformations are safe, effective, and reliable.


Surveying Manganese Oxides as Electrode Materials for Harnessing Salinity Gradient Energy.
The potential energy contained in the controlled mixing of waters with different salt concentrations (i.e., salinity gradient energy) can theoretically provide a substantial fraction of the global electrical demand. One method for generating electricity from salinity gradients is to use electrode-based reactions in electrochemical cells. Here, we examined the relationship between the electrical power densities generated from synthetic NaCl solutions and the crystal structures and morphologies of manganese oxides, which undergo redox reactions coupled to sodium ion uptake and release. Our aim was to make progress toward developing rational frameworks for selecting electrode materials used to harvest salinity gradient energy. We synthesized 12 manganese oxides having different crystal structures and particle sizes and measured the power densities they produced in a concentration flow cell fed with 0.02 and 0.5 M NaCl solutions. Power production varied considerably among the oxides, ranging from no power produced (β-MnO2) to 1.18 ± 0.01 W/m2 (sodium manganese oxide). Power production correlated with the materials' specific capacities, suggesting that cyclic voltammetry may be a simple method to screen possible materials. The highest power densities were achieved with manganese oxides capable of intercalating sodium ions when their potentials were prepoised prior to power production.


Impaired holistic processing in congenital prosopagnosia.
It has long been argued that face processing requires disproportionate reliance on holistic or configural processing, relative to that required for non-face object recognition, and that a disruption of such holistic processing may be causally implicated in prosopagnosia. Previously, we demonstrated that individuals with congenital prosopagnosia (CP) did not show the normal face inversion effect (better performance for upright compared to inverted faces) and evinced a local (rather than the normal global) bias in a compound letter global/local (GL) task, supporting the claim of disrupted holistic processing in prosopagnosia. Here, we investigate further the nature of holistic processing impairments in CP, first by confirming, in a large sample of CP individuals, the absence of the normal face inversion effect and the presence of the local bias on the GL task, and, second, by employing the composite face paradigm, often regarded as the gold standard for measuring holistic face processing. In this last task, we show that, in contrast with controls, the CP group perform equivalently with aligned and misaligned faces and was impervious to (the normal) interference from the task-irrelevant bottom part of faces. Interestingly, the extent of the local bias evident in the composite task is correlated with the abnormality of performance on diagnostic face processing tasks. Furthermore, there is a significant correlation between the magnitude of the local bias in the GL and performance on the composite task. These results provide further evidence for impaired holistic processing in CP and, moreover, corroborate the critical role of this type of processing for intact face recognition.


Single-row vs. double-row arthroscopic rotator cuff repair: clinical and 3 Tesla MR arthrography results.
Arthroscopic rotator cuff repair has become popular in the last few years because it avoids large skin incisions and deltoid detachment and dysfunction. Earlier arthroscopic single-row (SR) repair methods achieved only partial restoration of the original footprint of the tendons of the rotator cuff, while double-row (DR) repair methods presented many biomechanical advantages and higher rates of tendon-to-bone healing. However, DR repair failed to demonstrate better clinical results than SR repair in clinical trials. MR imaging at 3 Tesla, especially with intra-articular contrast medium (MRA), showed a better diagnostic performance than 1.5 Tesla in the musculoskeletal setting. The objective of this study was to retrospectively evaluate the clinical and 3 Tesla MRA results in two groups of patients operated on for a medium-sized full-thickness rotator cuff tear with two different techniques. The first group consisted of 20 patients operated on with the SR technique; the second group consisted of 20 patients operated on with the DR technique. All patients were evaluated at a minimum of 3 years after surgery. The primary end point was the re-tear rate at 3 Tesla MRA. The secondary end points were the Constant-Murley Scale (CMS), the Simple Shoulder Test (SST) scores, surgical time and implant expense. The mean follow-up was 40 months in the SR group and 38.9 months in the DR group. The mean postoperative CMS was 70 in the SR group and 68 in the DR group. The mean SST score was 9.4 in the SR group and 10.1 in the DR group. The re-tear rate was 60% in the SR group and 25% in the DR group. Leakage of the contrast medium was observed in all patients. To the best of our knowledge, this is the first report on 3 Tesla MRA in the evaluation of two different techniques of rotator cuff repair. DR repair resulted in a statistically significant lower re-tear rate, with longer surgical time and higher implant expense, despite no difference in clinical outcomes. We think that leakage of the contrast medium is due to an incomplete tendon-to-bone sealing, which is not a re-tear. This phenomenon could have important medicolegal implications. Level of evidence III. Treatment study: Case-control study.


[Gene therapy as a treatment concept for inherited retinal diseases].
Gene therapy for inherited retinal diseases (IRDs) is currently being validated in several clinical trials and is becoming a promising therapeutic option for these previously incurable diseases. The aim of this review is to give an overview of the concept, the application and the challenges associated with gene therapy. In particular, the pertinence of gene therapy for IRDs will be highlighted along with ongoing clinical trials in the field. A systematic review of relevant entries on gene therapy and on gene therapy for IRDs, in particular in PubMed and ClinicalTrials.gov. Gene therapy is emerging not only as a therapy for monogenetic retinal diseases but also for complex genetic diseases, such as neovascular age-related macular degeneration. The discovery of adeno-associated viral vectors (AAVs) has marked a great improvement for IRD gene therapy. All clinical studies since 2006 demonstrated the safety and initial efficacy; however, not all expectations based on very successful preclinical studies were met. In future we can expect gene therapy to continue to become more clinically relevant. More than ever, it is now essential to generate precise characterizations of the natural disease progression of IRDs through observational or retrospective studies in order to guarantee a most effective study design.


Tissue distribution of P-glycoprotein encoded by a multidrug-resistant gene as revealed by a monoclonal antibody, MRK 16.
A monoclonal antibody, MRK 16, specific to a human myelogenous leukemia cell line, K-562, and resistant to Adriamycin, was used to determine the localization of the antigen molecules (P-glycoprotein) recognized by the monoclonal antibody. P-glycoprotein was found to be expressed very strongly in the adrenal cortex and medulla of adults and strongly in the renal tubules of the kidney and the placenta. Interestingly, P-glycoprotein was not distributed in fetal and neonatal adrenals, and thus may be closely related to adrenal maturation. A high level of P-glycoprotein expression was also seen in one case each of untreated lung cancer (one of ten) and breast cancer (one of nine). Immunoelectron microscopically, the P-glycoprotein was distributed evenly on the membranes of K-562/ADM and 2780 cells. These results imply that the presence of the glycoprotein may be useful as a marker for in vitro studies of multidrug resistance in various malignancies and as an indicator of therapeutic efficacy of ex vivo eradication of multidrug-resistant cancer cells, although other mechanisms of drug resistance may exist, and there is a possibility that this MRK 16 monoclonal antibody may not recognize all P-glycoprotein.


Improving MEME via a two-tiered significance analysis.
With over 9000 unique users recorded in the first half of 2013, MEME is one of the most popular motif-finding tools available. Reliable estimates of the statistical significance of motifs can greatly increase the usefulness of any motif finder. By analogy, it is difficult to imagine evaluating a BLAST result without its accompanying E-value. Currently MEME evaluates its EM-generated candidate motifs using an extension of BLAST's E-value to the motif-finding context. Although we previously indicated the drawbacks of MEME's current significance evaluation, we did not offer a practical substitute suited for its needs, especially because MEME also relies on the E-value internally to rank competing candidate motifs. Here we offer a two-tiered significance analysis that can replace the E-value in selecting the best candidate motif and in evaluating its overall statistical significance. We show that our new approach could substantially improve MEME's motif-finding performance and would also provide the user with a reliable significance analysis. In addition, for large input sets, our new approach is in fact faster than the currently implemented E-value analysis.


In Vitro Efficacy of Six Alternative Antibiotics against Multidrug Resistant Escherichia Coli and Klebsiella Pneumoniae from Urinary Tract Infections.
Increasing resistance in Escherichia coli and Klebsiella pneumoniae to firstline antibiotics makes therapeutic options for urinary tract infections (UTIs) challenging. This study investigated the in vitro efficacies of 6 antibiotics against multidrug resistant (MDR) uropathogens. Minimum inhibitory concentrations to ceftibuten, cefpodoxime, fosfomycin, mecillinam, temocillin, and trimethoprim were determined against 155 MDR-isolates of E. coli and K. pneumoniae. The presence of extended-spectrum beta-lactamases (ESBL) and plasmid-borne AmpC enzymes was determined by phenotypic testing with genotyping performed by multiplex polymerase chain reaction. Temocillin demonstrated highest susceptibility rates for both E. coli (95%) and K. pneumoniae (95%) when breakpoints for uncomplicated UTIs were applied; however, temocillin susceptibility was substantially lower when "systemic infection" breakpoints were used. Fosfomycin demonstrated the best in vitro efficacy of the orally available agents, with 78% and 69% of E. coli and K. pneumoniae isolates susceptible, respectively. The next most effective antibiotics were ceftibuten (45%) and mecillinam (32%). ESBL and ampC genes were present in 47 (30%) and 59 (38%) isolates. This study demonstrated few oral therapeutic options for MDR-uropathogens, with fosfomycin demonstrating the best in vitro activity.


Behavioral phenotypes of inbred mouse strains: implications and recommendations for molecular studies.
Choosing the best genetic strains of mice for developing a new knockout or transgenic mouse requires extensive knowledge of the endogenous traits of inbred strains. Background genes from the parental strains may interact with the mutated gene, in a manner which could severely compromise the interpretation of the mutant phenotype. The present overview summarizes the literature on a wide variety of behavioral traits for the 129, C57BL/6, DBA/2, and many other inbred strains of mice. Strain distributions are described for open field activity, learning and memory tasks, aggression, sexual and parental behaviors, acoustic startle and prepulse inhibition, and the behavioral actions of ethanol, nicotine, cocaine, opiates, antipsychotics, and anxiolytics. Using the referenced information, molecular geneticists can choose optimal parental strains of mice, and perhaps develop new embryonic stem cell progenitors, for new knockouts and transgenics to investigate gene function, and to serve as animal models in the development of novel therapeutics for human genetic diseases.


Furcation anatomy of the first mandibular molar in dogs.
Tooth surface and interradicular area (furcation) measurements were taken of 20 first mandibular molar teeth obtained randomly from canine cadavers. The lingual furcation entrance had a mean width of 1.2 +/- 0.3 mm. The buccal furcation entrance had a mean width of 1.3 +/- 0.4 mm. There was no significant difference between mean furcation entrance measurements. The horizontal attachment area between the cementoenamel junction and the coronal roof of the lingual furcation was 1.1 +/- 0.4 mm. The horizontal attachment area between the cementoenamel junction and the coronal roof of the buccal furcation (0.5 +/- 0.3 mm) was significantly different from the lingual horizontal attachment area. Distal root length and mesial root length were 16.0 +/- 1.5 mm and 16.1 +/- 1.6 mm, respectively. The mean coefficient of variation for variables measured was 14.0%. Tooth size did not have a significant effect on furcation entrance measurement. All teeth had a concavity coronal to the furcation area, which extended apically for a mean distance of 65.6% of the distal root length and 83.8% of the mesial root length. Furcation anatomy of the first mandibular molar is complex and may be a contributing factor in periodontal disease involving the interradicular area.


Appearance and Frequency of Gas Interface Artifacts Involving Small Bowel on Rapid-Voltage-Switching Dual-Energy CT Iodine-Density Images.
The purpose of this study is to describe the appearance and frequency of gas interface artifacts in the jejunum that may mimic severe bowel disease on iodine-density images generated from rapid-voltage-switching dual-energy CT (DECT) scans. Two readers retrospectively reviewed 108 consecutive abdominal rapid-voltage-switching DECT scans to record the presence of image artifacts in jejunal segments with different degrees of gaseous luminal filling, classified as full, partial, or absent. Readers viewed iodine-density images and corresponding 140-kVp and 65-keV virtual monochromatic images and classified the jejunal artifacts on iodine-density images as pseudostratified appearance of the bowel wall, pseudopneumatosis, pseudohyperenhancement, or pseudohypoenhancement. We correlated the presence of the artifacts with clinical features suggesting bowel disease. Image artifacts were found in 91 of 108 scans (84.3%), appeared in 148 of 265 jejunal segments (55.8%), and included each type except for pseudohypoenhancement. Artifacts occurred exclusively when gas was present in the bowel lumen and were seen in 59 of 59 (100%) fully gas-distended segments, 89 of 98 (90.8%) partially gas-distended segments, and none of 108 gas-absent segments (p < 0.0001). In fully and partially gas-distended jejunal segments (n = 157), 148 (94.3%) segments had two or more artifacts. None of the patients was found to have clinical bowel-related injury on follow-up of medical records. Pseudostratified appearance, pseudopneumatosis, and pseudohyperenhancement, but not pseudohypoenhancement, artifacts are common in gas-filled jejunal segments on iodine-density images generated from rapid-voltage-switching DECT scans and are not seen in the corresponding 140-kVp or 65-keV images. Knowledge of the appearance of such iodine-density image artifacts will avoid potential examination interpretation pitfalls.


Kin selection may contribute to lek evolution and trait introgression across an avian hybrid zone.
Understanding the mechanism(s) that favour cooperation among individuals competing for the same resources provides direct insights into the evolution of grouping behaviour. In a hybrid zone between golden-/yellow-collared (Manacus vitellinus) and white-collared (Manacus candei) manakins, males form aggregations composed of white and yellow males solely to attract females ('mixed leks'). Previous work shows that yellow males in these mixed leks experience a clear mating advantage over white males, resulting in the preferential introgression of yellow plumage allele(s) into the white species. However, the yellow male mating advantage only occurs in mixed leks with high frequencies of yellow males, and only a few of these males probably mate. Hence, it remains unclear why unsuccessful males join leks. Here, we used microsatellite markers to estimate pairwise relatedness among males within and between leks to test whether indirect genetic benefits of helping kin ('kin selection') can promote grouping. We found that yellow males are significantly more related to each other within than between leks, while relatedness among white males did not differ within and between leks. This suggests that yellow males may indirectly enhance their own reproductive success by preferentially lekking with relatives because yellow plumage is under positive frequency-dependent selection (positive FDS). Our results are consistent with the hypothesis that kin selection may promote grouping and facilitate positive FDS for yellow males, mediating the movement of yellow plumage across this hybrid zone.


Active yeast ribosome preparation using monolithic anion exchange chromatography.
In vitro studies of translation provide critical mechanistic details, yet purification of large amounts of highly active eukaryotic ribosomes remains a challenge for biochemists and structural biologists. Here, we present an optimized method for preparation of highly active yeast ribosomes that could easily be adapted for purification of ribosomes from other species. The use of a nitrogen mill for cell lysis coupled with chromatographic purification of the ribosomes results in 10-fold-increased yield and less variability compared with the traditional approach, which relies on sedimentation through sucrose cushions. We demonstrate that these ribosomes are equivalent to those made using the traditional method in a host of in vitro assays, and that utilization of this new method will consistently produce high yields of active yeast ribosomes.


Immunological and toxicological implications of short-term studies in animals of pharmaceutical aerosol delivery to the lungs: relevance to humans.
The effects of xenobiotics on the lungs have been studied for many years. In the past 50 years, delivery of drugs to the lungs has been adopted to achieve local effects, specifically for the treatment of asthma. Recently, due to the proximity of the circulating blood supply and to their large surface area, the lungs have been proposed as the port of entry for drugs to obtain systemic effects, particularly for macromolecular compounds of biological origin. Numerous studies regarding drug formulation, delivery systems, and related pharmacokinetics have been reported; however, the concurrent effects of pulmonary delivery of drugs on the physiology of the lung has not been evaluated early in the development process. The prospect of using the lungs for the delivery of biological molecules such as proteins, peptides, and nucleic acids raises the question of the local toxicity of these compounds. Therefore, criteria must be established to study the initial impact of pulmonary drug delivery on the physiology of the lungs. This relates particularly to subtle local and systemic implications of those effects on the transport phenomena that may be contrasted with conventional toxicity studies focused on gross effects.


Prediction of intracellular metabolic states from extracellular metabolomic data.
Metabolic models can provide a mechanistic framework to analyze information-rich omics data sets, and are increasingly being used to investigate metabolic alternations in human diseases. An expression of the altered metabolic pathway utilization is the selection of metabolites consumed and released by cells. However, methods for the inference of intracellular metabolic states from extracellular measurements in the context of metabolic models remain underdeveloped compared to methods for other omics data. Herein, we describe a workflow for such an integrative analysis emphasizing on extracellular metabolomics data. We demonstrate, using the lymphoblastic leukemia cell lines Molt-4 and CCRF-CEM, how our methods can reveal differences in cell metabolism. Our models explain metabolite uptake and secretion by predicting a more glycolytic phenotype for the CCRF-CEM model and a more oxidative phenotype for the Molt-4 model, which was supported by our experimental data. Gene expression analysis revealed altered expression of gene products at key regulatory steps in those central metabolic pathways, and literature query emphasized the role of these genes in cancer metabolism. Moreover, in silico gene knock-outs identified unique control points for each cell line model, e.g., phosphoglycerate dehydrogenase for the Molt-4 model. Thus, our workflow is well-suited to the characterization of cellular metabolic traits based on extracellular metabolomic data, and it allows the integration of multiple omics data sets into a cohesive picture based on a defined model context.


SARS-CoV-2 Viral Entry Proteins in Hyperandrogenemic Female Mice: Implications for Women with PCOS and COVID-19.
SARS-CoV-2, the causative agent of COVID-19, infects host cells using the angiotensin I converting enzyme 2 (ACE2) as its receptor after priming by host proteases, including TMPRSS2. COVID-19 affects multiple organ systems, and male patients suffer increased severity and mortality. Polycystic Ovary Syndrome (PCOS) is the most common endocrine disorder in reproductive-age women and is characterized by hyperandrogenism, ovulatory dysfunction, and polycystic ovarian morphology. PCOS is associated with obesity and cardiometabolic comorbidities, both being risk factors associated with severe COVID-19 pathology. We hypothesize that elevated androgens in PCOS regulate SARS-CoV-2 entry proteins in multiple tissues increasing the risk for this population. Female mice were treated with dihydrotestosterone (DHT) for 90 days. Body composition was measured by EchoMRI. Fasting glucose was determined by an enzymatic method. mRNA and protein levels of ACE2, Tmprss2, Cathepsin L, Furin, Tmprss4, and Adam17 were quantified by RT-qPCR, Western-blot, or ELISA in tissues, serum, and urine. DHT treatment increased body weight, fat and lean mass, and fasting glucose. Ace2 mRNA was upregulated in the lung, cecum, heart, and kidney, while downregulated in the brain by DHT. ACE2 protein was upregulated by DHT in the small intestine, heart, and kidney. The SARS-CoV-2 priming proteases Tmprss2, Cathepsin L, and Furin mRNA were upregulated by DHT in the kidney. ACE2 sheddase Adam17 mRNA was upregulated by DHT in the kidney, which corresponded with increased urinary ACE2 in DHT treated mice. Our results highlight the potential for increased cardiac, renal, and gastrointestinal dysfunction in PCOS women with COVID-19.


Loss of TFF1 is associated with activation of NF-κB-mediated inflammation and gastric neoplasia in mice and humans.
Trefoil factor 1 (TFF1) is a tumor suppressor gene that encodes a peptide belonging to the trefoil factor family of protease-resistant peptides. Although TFF1 expression is frequently lost in gastric carcinomas, the tumorigenic pathways this affects have not been determined. Here we show that Tff1-knockout mice exhibit age-dependent carcinogenic histological changes in the pyloric antrum of the gastric mucosa, progressing from gastritis to hyperplasia, low-grade dysplasia, high-grade dysplasia, and ultimately malignant adenocarcinoma. The histology and molecular signatures of gastric lesions in the Tff1-knockout mice were consistent with an inflammatory phenotype. In vivo, ex-vivo, and in vitro studies showed that TFF1 expression suppressed TNF-α-mediated NF-κB activation through the TNF receptor 1 (TNFR1)/IκB kinase (IKK) pathway. Consistent with these mouse data, human gastric tissue samples displayed a progressive decrease in TFF1 expression and an increase in NF-κB activation along the multi-step carcinogenesis cascade. Collectively, these results provide evidence that loss of TFF1 leads to activation of IKK complex-regulated NF-κB transcription factors and is an important event in shaping the NF-κB-mediated inflammatory response during the progression to gastric tumorigenesis.


Brown adipose tissue is the key depot for glucose clearance in microbiota depleted mice.
Gut microbiota deficient mice demonstrate accelerated glucose clearance. However, which tissues are responsible for the upregulated glucose uptake remains unresolved, with different studies suggesting that browning of white adipose tissue, or modulated hepatic gluconeogenesis, may be related to enhanced glucose clearance when the gut microbiota is absent. Here, we investigate glucose uptake in 22 different tissues in 3 different mouse models. We find that gut microbiota depletion via treatment with antibiotic cocktails (ABX) promotes glucose uptake in brown adipose tissue (BAT) and cecum. Nevertheless, the adaptive thermogenesis and the expression of uncoupling protein 1 (UCP1) are dispensable for the increased glucose uptake and clearance. Deletion of Ucp1 expressing cells blunts the improvement of glucose clearance in ABX-treated mice. Our results indicate that BAT and cecum, but not white adipose tissue (WAT) or liver, contribute to the glucose uptake in the gut microbiota depleted mouse model and this response is dissociated from adaptive thermogenesis.


Increased reaction time variability in attention-deficit hyperactivity disorder as a response-related phenomenon: evidence from single-trial event-related potentials.
Increased intra-subject variability (ISV) in reaction times (RTs) is a promising endophenotype for attention-deficit hyperactivity disorder (ADHD) and among the most robust hallmarks of the disorder. ISV has been assumed to represent an attentional deficit, either reflecting lapses in attention or increased neural noise. Here, we use an innovative single-trial event-related potential approach to assess whether the increased ISV associated with ADHD is indeed attributable to attention, or whether it is related to response-related processing. We measured electroencephalographic responses to working memory oddball tasks in patients with ADHD (N = 20, aged 11.3 ± 1.1) and healthy controls (N = 25, aged 11.7 ± 1.1), and analysed these data with a recently developed method of single-trial event-related potential analysis. Estimates of component latency variability were computed for the stimulus-locked and response-locked forms of the P3b and the lateralised readiness potential (LRP). ADHD patients showed significantly increased ISV in behavioural ISV. This increased ISV was paralleled by an increase in variability in response-locked event-related potential latencies, while variability in stimulus-locked latencies was equivalent between groups. This result held across the P3b and LRP. Latency of all components predicted RTs on a single-trial basis, confirming that all were relevant for speed of processing. These data suggest that the increased ISV found in ADHD could be associated with response-end, rather than stimulus-end processes, in contrast to prevailing conceptions about the endophenotype. This mental chronometric approach may also be useful for exploring whether the existing lack of specificity of ISV to particular psychiatric conditions can be improved upon.


Cost-effective management alternatives for Snake River Chinook salmon: a biological-economic synthesis.
The mandate to increase endangered salmon populations in the Columbia River Basin of North America has created a complex, controversial resource-management issue. We constructed an integrated assessment model as a tool for analyzing biological-economic trade-offs in recovery of Snake River spring- and summer-run chinook salmon (Oncorhynchus tshawytscha). We merged 3 frameworks: a salmon-passage model to predict migration and survival of smolts; an age-structured matrix model to predict long-term population growth rates of salmon stocks; and a cost-effectiveness analysis to determine a set of least-cost management alternatives for achieving particular population growth rates. We assessed 6 individual salmon-management measures and 76 management alternatives composed of one or more measures. To reflect uncertainty, results were derived for different assumptions of effectiveness of smolt transport around dams. Removal of an estuarine predator, the Caspian Tern (Sterna caspia), was cost-effective and generally increased long-term population growth rates regardless of transport effectiveness. Elimination of adult salmon harvest had a similar effect over a range of its cost estimates. The specific management alternatives in the cost-effective set depended on assumptions about transport effectiveness. On the basis of recent estimates of smolt transport effectiveness, alternatives that discontinued transportation or breached dams were prevalent in the cost-effective set, whereas alternatives that maximized transportation dominated if transport effectiveness was relatively high. More generally, the analysis eliminated 80-90% of management alternatives from the cost-effective set. Application of our results to salmon management is limited by data availability and model assumptions, but these limitations can help guide research that addresses critical uncertainties and information. Our results thus demonstrate that linking biology and economics through integrated models can provide valuable tools for science-based policy and management.


mTOR: a pharmacologic target for autophagy regulation.
mTOR, a serine/threonine kinase, is a master regulator of cellular metabolism. mTOR regulates cell growth and proliferation in response to a wide range of cues, and its signaling pathway is deregulated in many human diseases. mTOR also plays a crucial role in regulating autophagy. This Review provides an overview of the mTOR signaling pathway, the mechanisms of mTOR in autophagy regulation, and the clinical implications of mTOR inhibitors in disease treatment.


Are patients better than the laboratory in assessing sweating? Validation study.
Primary focal hyperhidrosis is a hereditary condition characterized by excessive sweating affecting a variety of areas, including the palms. In most cases, the decision to treat a patient is based only on the patient's own description of the symptoms because laboratory tests have not been established in clinical practice. To validate a subjective self-assessment scale for sweating in comparison with an objective method of quantifying evaporation from the skin surface. We studied palmar sweating in 20 patients with primary focal hyperhidrosis and 20 age- and gender-matched controls. Four times in each subject, we obtained measurements of the evaporation in palms and the self-evaluation scale (SES) scores on a proportional scale from 0 (no sweating) to 10 (worst imaginable sweating). Good correlation was found between the SES score and evaporation assessment (R = .65). The SES score was easier to obtain and provided better diagnostic sensitivity and specificity. They leveled out at over 80% when a score of 3 was used as the threshold level; for evaporimetry, sensitivity equalled specificity at 70% at the threshold 90 mg -2h(-1). We confirm the usefulness of the SES in assessing palmar sweating and discuss the current restrictions in its use in clinical diagnosis. We propose that the scale can be added effortlessly to research or clinical protocols to provide the necessary validation in wider spectra of patients.


A Study of Traveller Horse Owners' Attitudes to Horse Care and Welfare Using an Equine Body Condition Scoring System.
Traveller horses are often perceived to be exposed to poor welfare due to Travellers' traditional way of horsemanship. However, few studies have investigated Traveller horse welfare. Hence, the present study aims to explore Traveller horse owners' attitudes to horse care and welfare. Semi-structured interviews and discussion groups examined 14 Irish Traveller horse owners' attitudes and approach to horse ownership. Additionally, a body condition scoring (BCS) instrument was assessed for its accuracy and ease of use when applied by Traveller horse owners. Additionally, the BCS system was used to assess 18 horses. Results show that Travellers have a good understanding of horses' natural behaviours and environment, which is reflected in their management practices. However, barriers to improved welfare are land availability, since landowners are often reluctant to lease to Travellers, and the impoundment of horses as a consequence of fly grazing, under the Control of Horses Act 1996 (Ireland). Furthermore, Travellers regarded the BCS as a useful tool, but would require training to apply the scoring successfully. The results suggest that attitudes and management practices are favourable, but Travellers have limited means to overcome barriers. Therefore, it is necessary to increase capacity building and assist with the acquisition of land.


Protein-nanoparticle interactions and a new insight.
The study of protein-nanoparticle interactions provides knowledge about the bio-reactivity of nanoparticles, and creates a database of nanoparticles for applications in nanomedicine, nanodiagnosis, and nanotherapy. The problem arises when nanoparticles come in contact with physiological fluids such as plasma or serum, wherein they interact with the proteins (or other biomolecules). This interaction leads to the coating of proteins on the nanoparticle surface, mostly due to the electrostatic interaction, called 'corona'. These proteins are usually partially unfolded. The protein corona can deter nanoparticles from their targeted functionalities, such as drug/DNA delivery at the site and fluorescence tagging of diseased tissues. The protein corona also has many repercussions on cellular intake, inflammation, accumulation, degradation, and clearance of the nanoparticles from the body depending on the exposed part of the proteins. Hence, the protein-nanoparticle interaction and the configuration of the bound-proteins on the nanosurface need thorough investigation and understanding. Several techniques such as DLS and zeta potential measurement, UV-vis spectroscopy, fluorescence spectroscopy, circular dichroism, FTIR, and DSC provide valuable information in the protein-nanoparticle interaction study. Besides, theoretical simulations also provide additional understanding. Despite a lot of research publications, the fundamental question remained unresolved. Can we aim for the application of functional nanoparticles in medicine? A new insight, given by us, in this article assumes a reasonable solution to this crucial question.


Immigrants in the one percent: The national origin of top wealth owners.
Economic inequality in the United States is extreme, but little is known about the national origin of affluent households. Households in the top one percent by total wealth own vastly disproportionate quantities of household assets and have correspondingly high levels of economic, social, and political influence. The overrepresentation of white natives (i.e., those born in the U.S.) among high-wealth households is well-documented, but changing migration dynamics suggest that a growing portion of top households may be immigrants. Because no single survey dataset contains top wealth holders and data about country of origin, this paper uses two publicly-available data sets: the Survey of Consumer Finances (SCF) and the Survey of Income and Program Participation (SIPP). Multiple imputation is used to impute country of birth from the SIPP into the SCF. Descriptive statistics are used to demonstrate reliability of the method, to estimate the prevalence of immigrants among top wealth holders, and to document patterns of asset ownership among affluent immigrants. Significant numbers of top wealth holders who are usually classified as white natives may be immigrants. Many top wealth holders appear to be European and Canadian immigrants, and increasing numbers of top wealth holders are likely from Asia and Latin America as well. Results suggest that of those in the top one percent of wealth holders, approximately 3% are European and Canadian immigrants, .5% are from Mexico or Cuban, and 1.7% are from Asia (especially Hong Kong, Taiwan, Mainland China, and India). Ownership of key assets varies considerably across affluent immigrant groups. Although the percentage of top wealth holders who are immigrants is relatively small, these percentages represent large numbers of households with considerable resources and corresponding social and political influence. Evidence that the propensity to allocate wealth to real and financial assets varies across immigrant groups suggests that wealth ownership is more global than previous research suggests and that immigrant groups are likely to become more prevalent in top wealth positions in the U.S. As the representation of immigrants in top wealth positions grows, their economic, social, and political influence is likely to increase as well.


The phylogenetic affinities of the bizarre Late Cretaceous Romanian theropod Balaur bondoc (Dinosauria, Maniraptora): dromaeosaurid or flightless bird?
The exceptionally well-preserved Romanian dinosaur Balaur bondoc is the most complete theropod known to date from the Upper Cretaceous of Europe. Previous studies of this remarkable taxon have included its phylogenetic interpretation as an aberrant dromaeosaurid with velociraptorine affinities. However, Balaur displays a combination of both apparently plesiomorphic and derived bird-like characters. Here, we analyse those features in a phylogenetic revision and show how they challenge its referral to Dromaeosauridae. Our reanalysis of two distinct phylogenetic datasets focusing on basal paravian taxa supports the reinterpretation of Balaur as an avialan more crownward than Archaeopteryx but outside of Pygostylia, and as a flightless taxon within a paraphyletic assemblage of long-tailed birds. Our placement of Balaur within Avialae is not biased by character weighting. The placement among dromaeosaurids resulted in a suboptimal alternative that cannot be rejected based on the data to hand. Interpreted as a dromaeosaurid, Balaur has been assumed to be hypercarnivorous and predatory, exhibiting a peculiar morphology influenced by island endemism. However, a dromaeosaurid-like ecology is contradicted by several details of Balaur's morphology, including the loss of a third functional manual digit, the non-ginglymoid distal end of metatarsal II, and a non-falciform ungual on the second pedal digit that lacks a prominent flexor tubercle. Conversely, an omnivorous ecology is better supported by Balaur's morphology and is consistent with its phylogenetic placement within Avialae. Our reinterpretation of Balaur implies that a superficially dromaeosaurid-like taxon represents the enlarged, terrestrialised descendant of smaller and probably volant ancestors.


Introduction of the blood pressure cuff into U.S. medical practice: technology and skilled practice.
The history of the sphygmomanometer, or blood pressure cuff, raises useful questions about the acceptance of new medical technologies. When the blood pressure cuff first appeared in U.S. medical practice in the first decade of the 1900s, it generated some concern and debate among physicians. Review of the medical literature, a systematic study of patient records from Massachusetts General Hospital, and consideration of events in Boston during this period suggest that physicians faced several important choices associated with the early acceptance of the cuff. The introduction of the blood pressure cuff presented physicians with several different, competing methods for assessing the force of a patient's blood flow. Physicians chose to use the cuff in a manner that preserved their exclusive use of the new tool and maintained a high level of skill for their individual practices. An early proposal to introduce the new blood pressure cuff as a simple tool for nursing use met with resistance. Many physicians initially favored a competing practice of assessing the force of blood flow by pulse palpation. Physicians eventually dropped the practice of subjectively palpating the force of blood flow and came to rely increasingly on the measurement of blood pressure using auscultation. Even after adopting the cuff into practice, however, they had little interest in completely standardizing the use of the blood pressure cuff across the practices of individual physicians.


Immobilization of zinc phthalocyanines in silicate matrices and investigation of their photobactericidal effect on E. coli.
The aim of the present investigation was to immobilize zinc phthalocyanines in a silicate matrix and to test the photobactericidal properties of the matrices so prepared toward Esherichia coli in model aqueous media. For the purpose, tetra tertiary butyl zinc phthalocyanine (TBZnPc) and zinc phthalocyanine tetrasulfonic acid (ZnPcTS) were used. The abilities of these two photosensitizers to generate singlet oxygen in solution were compared by following the rate of photobleaching of 1,3-diphenylisobenzofuran (DPBF) at 430 nm in dimethylformamide (DMF). The results of this study show clearly that, under the conditions used here, the TBZnPc is the more effective generator of singlet oxygen; with it the DPBF was virtually completely photobleached in 4 min, while with the ZnPcTS under the same conditions, it took 12 min to reach this point. Glass conjugates with the two phthalocyanines were obtained by the sol-gel technique and were characterized by a well-defined color due to the phthalocyanine incorporated in the silicate matrix. Glasses with an intense, but inhomogeneous, green color were obtained when the tetrasulfonic derivative of the zinc phthalocyanine was used, while blue glasses of evenly distributed coloration were formed from the tetra tertiary butyl derivative. The ZnPcTS conjugate demonstrates more effective singlet oxygen evolution than is the case with the TBZnPc conjugate. These results are the opposite of those obtained for the free phthalocyanines in solution. The structural formulae of the compounds show that TBZnPc has a more pronounced hydrophobic character than the sulfonic derivative. In our view, the relative reactivities of the conjugates can be explained by the tetrasulfonic derivative being situated mainly in the surface parts of the glass matrix where the hydrophilic character is prevailing, while the tertiary butyl derivative is mainly present in the internal parts of the matrix as a result of which it is less accessible and therefore less active. The results obtained on the effect of zinc phthalocyanine conjugates on E. coli show a trend similar to that observed with singlet oxygen evolution shown. Thus, for the ZnPcTS conjugate, the log kill is 1.32 and for the TBZnPc conjugate, it is 0.98, in each case after 120 min. The results obtained show that phthalocyanines can be immobilized successfully in a silicate matrix and used for photodisinfection of microbially polluted waters. The silicate matrix has some advantages in comparison with other organic matrices. It is insoluble in water, resistant towards microorganisms, easy to fabricate, and might be developed successfully for the photodisinfection of water, e.g., in swimming pools and in other open water reservoirs.


Self-consistent evolution of magnetic fields and chiral asymmetry in the early universe.
We show that the evolution of magnetic fields in a primordial plasma, filled with standard model particles at temperatures T≳10 MeV, is strongly affected by the chiral anomaly-an effect previously neglected. Although reactions, equilibrating left and right electrons, are in thermal equilibrium for T≲80 TeV, a left-right asymmetry develops in the presence of strong magnetic fields. This results in magnetic helicity transfer from shorter to longer scales and lepton asymmetry present in the plasma until T~10 MeV, which may strongly affect many processes in the early Universe.


The use of Systemic Lupus Erythematosus Disease Activity Index-2000 to define active disease and minimal clinically meaningful change based on data from a large cohort of systemic lupus erythematosus patients.
To examine SLEDAI-2000 cut-off scores for definition of active SLE and to determine the sensitivity to change of SLEDAI-2000 for the assessment of SLE disease activity and minimal clinically meaningful changes in score. Data from two multi-centre studies were used in the analysis: in a cross-sectional and a longitudinal fashion. At every assessment, data were collected on SLEDAI-2000 and treatment. The cross-sectional analysis with receiver operating characteristic (ROC) curves was used to examine the appropriate SLEDAI-2000 score to define active disease and increase in therapy was the reference standard. In the longitudinal analysis, sensitivity to change of SLEDAI-2000 was assessed with multinomial logistic regression. ROC curves analysis was used to examine possible cut-points in score changes associated with change in therapy, and mean changes were estimated. In the cross-sectional analysis, the most appropriate cut-off scores for active disease were 3 or 4. In the longitudinal analysis, the best model for predicting treatment increase was with the change in SLEDAI-2000 score and the score from the previous visit as continuous variables. The use of cut-points was less predictive of treatment change than the use of continuous score. The mean difference in the change in SLEDAI-2000 scores, adjusted for prior score, between patients with treatment increase and those without was 2.64 (95% CI 2.16, 3.14). An appropriate SLEDAI-2000 score to define active disease is 3 or 4. SLEDAI-2000 index is sensitive to change. The use of SLEDAI-2000 as a continuous outcome is recommended for comparative purposes.