Cp*CoIII-Catalyzed Alkylation of Primary and Secondary C(sp3)-H Bonds of 8-Alkylquinolines with Maleimides.
The cobalt(III)-catalyzed C(sp3)-H bond alkylation of 8-methyl quinoline with maleimides is reported. In contrast to the rhodium-catalyzed method, in the current cobalt-catalyzed method, a catalytic amount of acid is used, and importantly, it is also applicable to secondary C(sp3)-H bond alkylation. The developed methodology is applicable for N-alkyl- and N-aryl-substituted maleimides and unsubstituted maleimides, and it also tolerates the variety of functional groups on the 8-methyl quinoline moiety. Atom-economy and high regioselectivity with good to excellent yields of the alkylated products under mild reaction conditions are important features of this method.


Open chromatin defined by DNaseI and FAIRE identifies regulatory elements that shape cell-type identity.
The human body contains thousands of unique cell types, each with specialized functions. Cell identity is governed in large part by gene transcription programs, which are determined by regulatory elements encoded in DNA. To identify regulatory elements active in seven cell lines representative of diverse human cell types, we used DNase-seq and FAIRE-seq (Formaldehyde Assisted Isolation of Regulatory Elements) to map "open chromatin." Over 870,000 DNaseI or FAIRE sites, which correspond tightly to nucleosome-depleted regions, were identified across the seven cell lines, covering nearly 9% of the genome. The combination of DNaseI and FAIRE is more effective than either assay alone in identifying likely regulatory elements, as judged by coincidence with transcription factor binding locations determined in the same cells. Open chromatin common to all seven cell types tended to be at or near transcription start sites and to be coincident with CTCF binding sites, while open chromatin sites found in only one cell type were typically located away from transcription start sites and contained DNA motifs recognized by regulators of cell-type identity. We show that open chromatin regions bound by CTCF are potent insulators. We identified clusters of open regulatory elements (COREs) that were physically near each other and whose appearance was coordinated among one or more cell types. Gene expression and RNA Pol II binding data support the hypothesis that COREs control gene activity required for the maintenance of cell-type identity. This publicly available atlas of regulatory elements may prove valuable in identifying noncoding DNA sequence variants that are causally linked to human disease.


Angiotensin II increases norepinephrine release from atria by acting on angiotensin subtype 1 receptors.
Norepinephrine stores in electrically driven guinea pig isolated atria were loaded with [3H]norepinephrine, and norepinephrine release was deduced from the radioactivity efflux. Electrical field stimulation of sympathetic nerve endings was applied during the refractory period of atrial contractions. The stimulation-induced release of norepinephrine was increased by angiotensin II (Ang II) (10(-8) to 10(-6) mol/L) in a concentration-dependent manner. The maximum observed effect was a 55% augmentation. The effects of 10(-7) and 10(-6) mol/L Ang II were abolished by 10(-6) and 10(-5) mol/L of the subtype 1 Ang II receptor antagonist losartan, respectively. Losartan by itself (10(-6) mol/L) caused a 14% reduction of norepinephrine release. The subtype 2 Ang II receptor ligand PD 123319 (1-[[4-(dimethylamino)-3-methylphenyl]methyl]-5-(diphenylacetyl)- 4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-6-carboxylic acid ditrifluoroacetate) in a concentration of 10(-4) mol/L had no detectable influence on transmitter release and did not antagonize the effect of Ang II. Angiotensin I (10(-6) and 10(-5) mol/L) increased norepinephrine release maximally by 23%. This effect was antagonized by 10(-5) mol/L losartan and did not appear in the presence of 10(-6) mol/L of the converting enzyme inhibitor ramiprilat. These results suggest that Ang II increases norepinephrine release by an activation of subtype 1 receptors, whereas angiotensin I is converted to Ang II to become effective.


Outbreaks of waterborne infectious intestinal disease in England and Wales, 1992-5.
Following the introduction of an improved surveillance system for infectious intestinal disease outbreaks in England and Wales, the Public Health Laboratory Service Communicable Disease Surveillance Centre received reports of 26 outbreaks between 1 January 1992 and 31 December 1995 in which there was evidence for waterborne transmission of infection. In these 26 outbreaks, 1756 laboratory confirmed cases were identified of whom 69 (4%) were admitted to hospital. In 19 outbreaks, illness was associated with the consumption of drinking water from public supplies (10 outbreaks) or private supplies (9 outbreaks). The largest outbreak consisted of 575 cases. In 4 of the remaining 7 outbreaks, illness was associated with exposure to swimming pool water. Cryptosporidium was identified as the probable causative organism in all 14 outbreaks associated with public water supplies and swimming pools. Campylobacter was responsible for most outbreaks associated with private water supplies. This review confirms a continuing risk of cryptosporidiosis from chlorinated water supplies in England and Wales, and reinforces governmental advice to water utilities that water treatment processes should be rigorously applied to ensure effective particle removal. High standards of surveillance are important for prompt recognition of outbreaks and institution of control measures. As microbiological evidence of water contamination may be absent or insufficient to implicate a particular water supply, a high standard of epidemiological investigation is recommended in all outbreaks of suspected waterborne disease.


Exercising alcohol patients don't lack motivation but struggle with structures, emotions and social context - a qualitative dropout study.
Exercise is an important component of a healthy lifestyle, the development of which is a relapse prevention strategy for those with alcohol use disorder. However, it is a challenge to create exercise interventions with a persistent behavioural change. The aim of this qualitative study was to investigate perceived barriers to participation in an exercise intervention among alcohol use disorder patients, who dropped out of the intervention program. Furthermore, this study aims to propose possibilities for a better practice of future intervention studies based on the participants' experiences and suggestions. Qualitative interviews with 17 patients who dropped out from an exercise intervention in an outpatient treatment centre about their experiences and reasons for dropping out. Social cognitive theory informed the development of the interview guides and systematic text condensation was used for analysis. Analysis revealed three central themes: 1) Structural barriers described as the type of exercise and the timing of the intervention, 2) Social barriers described as need for accountability and unsupportive relations, and 3) Emotional barriers described as fear, guilt and shame, and negative affect of the intervention on long term. Future exercise interventions should include socio-psychological support during the first weeks, begin shortly after treatment initiation instead of concurrently, and focus on garnering social support for participants in both the intervention context and among their existing network in order to best reduce barriers to participation. This study was retrospectively registered at Current Controlled Trials ISRCTN74889852 on 11 July 2013.


Extending drug ethno-epidemiology using agent-based modelling.
To show how the inclusion of agent-based modelling improved the integration of ethno-epidemiological data in a study of psychostimulant use and related harms among young Australians. Agent-based modelling, ethnographic fieldwork, in-depth interviews and epidemiological surveys. Melbourne, Perth and Sydney, Australia. Club drug users in Melbourne, recreational drug users in Perth and street-based injecting drug users in Sydney. Participants were aged 18-30 years and reported monthly or more frequent psychostimulant use. Agent-based modelling provided a specific focus for structured discussion about integrating ethnographic and epidemiological methods and data. The modelling process was underpinned by collective and incremental design principles, and produced 'SimAmph', a data-driven model of social and environmental agents and the relationships between them. Using SimAmph, we were able to test the probable impact of ecstasy pill-testing on the prevalence of harms--a potentially important tool for policy development. The study also navigated a range of challenges, including the need to manage epistemological differences, changes in the collective design process and modelling focus, the differences between injecting and non-injecting samples and concerns over the dissemination of modelling outcomes. Agent-based modelling was used to integrate ethno-epidemiological data on psychostimulant use, and to test the probable impact of a specific intervention on the prevalence of drug-related harms. It also established a framework for collaboration between research disciplines that emphasizes the synthesis of diverse data types in order to generate new knowledge relevant to the reduction of drug-related harms.


Altitude training considerations for the winter sport athlete.
Winter sports events routinely take place at low to moderate altitudes, and nearly all Winter Olympic Games have had at least one venue at an altitude >1000 m. The acute and chronic effects of altitude can have a substantial effect on performance outcomes. Acutely, the decline in oxygen delivery to working muscle decreases maximal oxygen uptake, negatively affecting performance in endurance events, such as cross-country skiing and biathlon. The reduction in air resistance at altitude can dramatically affect sports involving high velocities and technical skill components, such as ski jumping, speed skating, figure skating and ice hockey. Dissociation between velocity and sensations usually associated with work intensity (ventilation, metabolic signals in skeletal muscle and heart rate) may impair pacing strategy and make it difficult to determine optimal race pace. For competitions taking place at altitude, a number of strategies may be useful, depending on the altitude of residence of the athlete and ultimate competition altitude, as follows. First, allow extra time and practice (how much is yet undetermined) for athletes to adjust to the changes in projectile motion; hockey, shooting, figure skating and ski jumping may be particularly affected. These considerations apply equally in the reverse direction; that is, for athletes practising at altitude but competing at sea level. Second, allow time for acclimatization for endurance sports: 3-5 days if possible, especially for low altitude (500-2000 m); 1-2 weeks for moderate altitude (2000-3000 m); and at least 2 weeks if possible for high altitude (>3000 m). Third, increase exercise-recovery ratios as much as possible, with 1:3 ratio probably optimal, and consider more frequent substitutions for sports where this is allowed, such as ice hockey. Fourth, consider the use of supplemental O(2) on the sideline (ice hockey) or in between heats (skating and Alpine skiing) to facilitate recovery. For competitions at sea level, the 'live high-train low' model of altitude training can help athletes in endurance events to maximize performance.


The role of community acceptance over time for costs of HIV and STI prevention interventions: analysis of the Masaka Intervention Trial, Uganda, 1996-1999.
The objective of this study is to estimate the annual costs of information, education, and communication (IEC), both community- and school-based; strengthened public and private sexually transmitted infections treatment; condom social marketing (CSM); and voluntary counseling and testing (VCT) implemented in Masaka, Uganda, over 4 years, and to explore how unit costs change with varying population use/uptake. Total economic provider's costs and intervention outputs were collected annually to estimate annual unit costs between 1996 and 1999. In early intervention years, uptake of all activities grew dramatically and continued to grow for public STI treatment, CSM, and VCT. Attendance at IEC performances started to drop in year 4. Unit costs dropped rapidly with increasing uptake of and participation in interventions. When implementing long-term community-based interventions, it is important to take into account that it takes time for communities to scale up their participation, since this can lead to large variations in unit costs.


Short-term glucocorticoid treatment increases insulin secretion in islets derived from lean mice through multiple pathways and mechanisms.
Chronic exposure to elevated levels of glucocorticoids leads to metabolic dysfunctions with hyperglycemia and insulin resistance. Long-term treatment with glucocorticoids induces severe impairment of glucose-stimulated insulin secretion. We analyzed the effects of short-, and medium-term (2-120h) treatment with 50-200nM glucocorticoids on primary pancreatic islet cultures derived from lean C57BL/6J mice. In contrast to animal models of insulin resistance, beta-cells from lean mice respond with an increased glucose-stimulated insulin secretion, with a peak effect around 18-24h of treatment. Analyses of the insulin secretion response reveal that early and late phase responses are dissociated upon glucocorticoid treatment. Whereas late phase responses return to basal levels after long treatment, early phase responses remain increased over several days. Increased insulin secretion is also obtained by incubation with the inactive glucocorticoid dehydrocorticosterone, pointing to an important role of the enzyme 11beta-hydroxysteroid dehydrogenase type 1 in mediating glucocorticoid effects in beta-cells. Transcript profiling revealed differential regulation of genes involved in mediation of signal transduction, insulin secretion, stress and inflammatory responses. The results show that short- to medium-term glucocorticoid treatment of pancreatic islets derived from lean mice leads to an increased insulin release and may constitute an important parameter in changing towards a pro-diabetic phenotype.


24(S)-hydroxycholesterol participates in a liver X receptor-controlled pathway in astrocytes that regulates apolipoprotein E-mediated cholesterol efflux.
Both apolipoprotein E (apoE) and 24(S)-hydroxycholesterol are involved in the pathogenesis of Alzheimer disease (AD). It has been hypothesized that apoE affects AD development via isoform-specific effects on lipid trafficking between astrocytes and neurons. However, the regulation of the cholesterol supply of neurons via apoE-containing high density lipoproteins remains to be clarified. We show for the first time that the brain-specific metabolite of cholesterol produced by neurons, i.e. 24(S)-hydroxycholesterol, induces apoE transcription, protein synthesis, and secretion in a dose- and time-dependent manner in cells of astrocytic but not of neuronal origin. Moreover, 24(S)-hydroxycholesterol primes astrocytoma, but not neuroblastoma cells, to mediate cholesterol efflux to apoE. Similar results were obtained using the synthetic liver X receptor (LXR) agonist GW683965A, suggesting involvement of an LXR-controlled signaling pathway. A 10-20-fold higher basal LXRalpha and -beta expression level in astrocytoma compared with neuroblastoma cells may underlie these differential effects. Furthermore, apoE-mediated cholesterol efflux from astrocytoma cells may be controlled by the ATP binding cassette transporters ABCA1 and ABCG1, since their expression was also up-regulated by both compounds. In contrast, ABCG4 seems not to be involved, because its expression was induced only in neuronal cells. The expression of sterol regulatory element-binding protein (SREBP-2), low density lipoprotein receptor, 3-hydroxy-3-methylglutaryl-CoA reductase, and SREBP-1c was transiently up-regulated by GW683965A in astrocytes but down-regulated by 24(S)-hydroxycholesterol, suggesting that cholesterol efflux and synthesis are regulated independently. In conclusion, evidence is provided that 24(S)-hydroxycholesterol induces apoE-mediated efflux of cholesterol in astrocytes via an LXR-controlled pathway, which may be relevant for chronic and acute neurological diseases.


Validating a functional near-infrared spectroscopy diagnostic paradigm for Major Depressive Disorder.
Reduced haemodynamic response in the frontotemporal cortices of patients with major depressive disorder (MDD) has been demonstrated using functional near-infrared spectroscopy (fNIRS). Most notably, changes in cortical oxy-haemoglobin during a Japanese phonetic fluency task can differentiate psychiatric patients from healthy controls (HC). However, this paradigm has not been validated in the English language. Therefore, the present work aimed to distinguish patients with MDD from HCs, using haemodynamic response measured during an English letter fluency task. One hundred and five HCs and 105 patients with MDD took part in this study. NIRS signals during the verbal fluency task (VFT) was acquired using a 52-channel system, and changes in oxy-haemoglobin in the frontal and temporal regions were quantified. Depression severity, psychosocial functioning, pharmacotherapy and psychiatric history were noted. Patients with MDD had smaller changes in oxy-haemoglobin in the frontal and temporal cortices than HCs. In both regions of interest, oxy-haemoglobin was not associated with any of the clinical variables studied. 75.2% and 76.5% of patients with MDD were correctly classified using frontal and temporal region oxy-haemoglobin, respectively. Haemodynamic response measured by fNIRS during an English letter fluency task is a promising biomarker for MDD.


Integrated transcriptomic analysis of Trichosporon Asahii uncovers the core genes and pathways of fluconazole resistance.
Trichosporon asahii (T. asahii) has emerged as a dangerous pathogen that causes rare but life-threatening infections. Its resistance to certain antifungal agents makes it difficult to treat, especially for patients undergoing long-term antibiotic therapy. In this study, we performed a series of fluconazole (FLC) perturbation experiments for two T. asahii strains, a clinical isolate stain CBS 2479 (T2) and an environmental isolate strain CBS 8904 (T8), to uncover potential genes and pathways involved in FLC resistance. We achieved 10 transcriptomes of T2 and T8 that were based on dose and time series of FLC perturbations. Systematic comparisons of the transcriptomes revealed 32 T2 genes and 25 T8 genes that are highly sensitive to different FLC perturbations. In both T2 and T8 strains with the phenotype of FLC resistance, the processes of oxidation-reduction and transmembrane transport were detected to be significantly changed. The antifungal susceptibility testing of FLC and penicillin revealed their resistance pathways are merged. Accumulated mutations were found in 564 T2 and 225 T8 genes, including four highly mutated genes that are functionally related to the target of rapamycin complex (TOR). Our study provides abundant data towards genome-wide understanding of the molecular basis of FLC resistance in T. asahii.


[Mandatory treatment of forensic psychiatric patients in the Netherlands: costs and benefits in perspective].
So far, it is not known what costs and benefits are connected with the tbs-measure, a measure that involves a mandatory treatment programme for forensic psychiatric patients.
 AIM: To explore the costs and benefits that the tbs-measure has on society, on other important stakeholders such as victims and/or next-of-kin and the forensic psychiatric patients themselves.
 METHOD: We studied the relevant literature.
 RESULTS: The average costs of the tbs-treatment programme are 1.5 million euros. Additional costs result from recidivism among patients after tbs-treatment. Of these, 21.2% commit another serious offence after 9 years; this recidivism rate is much lower than rates for former offenders who have not received tbs-treatment (63.8%). Other costs arise through the impact that crimes have on stake-holders. Among the benefits of the tbs-programme are a reduction in psychopathological symptoms and in risk factors and lower recommitment rates (including judicial, non-judicial, voluntary and mandatory recommitment rates). Yet another benefit is the resultant increase of protective factors.
 CONCLUSION: Forensic psychiatric patients form a unique group within the mental health system in the Netherlands; these patients have multiple complex psychiatric problems and display serious criminal behavior. This group cannot easily be treated elsewhere in the existing judicial or mental health care system because these systems differ in (judicial) frameworks and have different treatment goals, and the forensic psychiatric patients have different psychiatric disorders and display more serious criminal behaviour than patients in the alternative systems. The daily costs of treatment in the tbs-system are higher that in other systems - but they are not exorbitant, given the complexity of the group. The tbs-measure therefore contributes to the safety of society.


Susceptibility of compound 48/80-sensitized Pseudomonas aeruginosa to the hydrophobic biocide triclosan.
Pseudomonas aeruginosa is intrinsically resistant to the hydrophobic biocide triclosan, and yet it can be sensitized to low concentrations by permeabilization of the outer membrane using compound 48/80. A selective plating assay revealed that compound 48/80-permeabilized YM64, a triclosan-recognizing efflux pump-deficient variant, was unable to initiate growth on a medium containing triclosan. Macrobroth dilution assay data revealed that treatment with compound 48/80 synergistically decreased minimal inhibitory concentrations of the hydrophobic antibacterial agents rifamycin SV and chloramphenicol for all cell envelope variant strains examined. A low concentration of triclosan exerted a transient bactericidal effect on permeabilized wild-type strain PAO1, after which exponential growth resumed within 4 h. Permeabilized strain YM64 was unable to overcome the inhibition; yet, both strains remained susceptible to chloramphenicol for as long as 6 h, thereby suggesting that the outer membrane remained permeable to nonpolar compounds. These data support the notion that the transitory nature of compound 48/80 sensitization to triclosan in P. aeruginosa does not involve obviation of the hydrophobic diffusion pathway through the outer membrane. The inability of strain YM64 to overcome the synergistic effect of compound 48/80 and triclosan strongly suggests that triclosan-recognizing efflux pumps are involved in maintaining viability in wild-type cells whose outer membranes are otherwise compromised.


Simulating Heterogeneous Tumor Cell Populations.
Certain tumor phenomena, like metabolic heterogeneity and local stable regions of chronic hypoxia, signify a tumor's resistance to therapy. Although recent research has shed light on the intracellular mechanisms of cancer metabolic reprogramming, little is known about how tumors become metabolically heterogeneous or chronically hypoxic, namely the initial conditions and spatiotemporal dynamics that drive these cell population conditions. To study these aspects, we developed a minimal, spatially-resolved simulation framework for modeling tissue-scale mixed populations of cells based on diffusible particles the cells consume and release, the concentrations of which determine their behavior in arbitrarily complex ways, and on stochastic reproduction. We simulate cell populations that self-sort to facilitate metabolic symbiosis, that grow according to tumor-stroma signaling patterns, and that give rise to stable local regions of chronic hypoxia near blood vessels. We raise two novel questions in the context of these results: (1) How will two metabolically symbiotic cell subpopulations self-sort in the presence of glucose, oxygen, and lactate gradients? We observe a robust pattern of alternating striations. (2) What is the proper time scale to observe stable local regions of chronic hypoxia? We observe the stability is a function of the balance of three factors related to O2-diffusion rate, local vessel release rate, and viable and hypoxic tumor cell consumption rate. We anticipate our simulation framework will help researchers design better experiments and generate novel hypotheses to better understand dynamic, emergent whole-tumor behavior.


Antibiotics promote inflammation through the translocation of native commensal colonic bacteria.
Antibiotic use is associated with an increased risk of developing multiple inflammatory disorders, which in turn are linked to alterations in the intestinal microbiota. How these alterations in the intestinal microbiota translate into an increased risk for inflammatory responses is largely unknown. Here we investigated whether and how antibiotics promote inflammation via the translocation of live native gut commensal bacteria. Oral antibiotics were given to wildtype and induced mutant mouse strains, and the effects on bacterial translocation, inflammatory responses and the susceptibility to colitis were evaluated. The sources of the bacteria and the pathways required for bacterial translocation were evaluated using induced mutant mouse strains, 16s rRNA sequencing to characterise the microbial communities, and in vivo and ex vivo imaging techniques. Oral antibiotics induced the translocation of live native commensal bacteria across the colonic epithelium, promoting inflammatory responses, and predisposing to increased disease in response to coincident injury. Bacterial translocation resulted from decreased microbial signals delivered to colonic goblet cells (GCs), was associated with the formation of colonic GC-associated antigen passages, was abolished when GCs were depleted and required CX3CR1(+) dendritic cells. Bacterial translocation occurred following a single dose of most antibiotics tested, and the predisposition for increased inflammation was only associated with antibiotics inducing bacterial translocation. These findings reveal an unexpected outcome of antibiotic therapy and suggest that bacterial translocation as a result of alterations in the intestinal microflora may provide a link between increasing antibiotic use and the increased incidence of inflammatory disorders.


Posttranslational glutamylation of alpha-tubulin.
The high degree of tubulin heterogeneity in neurons is controlled mainly at the posttranslational level. Several variants of alpha-tubulin can be posttranslationally labeled after incubation of cells with [3H]acetate or [3H]glutamate. Peptides carrying the radioactive moiety were purified by high-performance liquid chromatography. Amino acid analysis, Edman degradation sequencing, and mass spectrometric analysis of these peptides led to the characterization of a posttranslational modification consisting of the successive addition of glutamyl units on the gamma-carboxyl group of a glutamate residue (Glu445). This modification, localized within a region of alpha-tubulin that is important in the interactions of tubulin with microtubule-associated proteins and calcium, could play a role in regulating microtubule dynamics.


A comparison of face-to-face or internet-delivered physical activity intervention on targeted determinants.
This article describes the equivalency testing results of a 12-week behavior change program on targeted determinates of physical activity (PA) and self-reported health status. Participants (n = 192) were randomized to face-to-face, combined Internet and face-to-face, and Internet-only groups. Equivalency testing was used to examine differences and statistical equivalency across groups for all outcome measures (social support, self-efficacy, perceived health status, and motivational readiness for PA). Participants were assessed at baseline, postintervention, and 2 and 5 months postintervention. Motivational readiness for PA increased across all groups. The face-to-face and combined groups showed changes in social support; however, they were not statistically different and were equivalent. There were no changes in self-efficacy or physical health status. Overall face-to-face and the Internet delivery modes show similar results. If Internet-based programs can be shown to be as effective as face-to-face, they may in turn be a more efficient and cost-effective delivery method.


ECM-dependent HIF induction directs trophoblast stem cell fate via LIMK1-mediated cytoskeletal rearrangement.
The Hypoxia-inducible Factor (HIF) family of transcriptional regulators coordinates the expression of dozens of genes in response to oxygen deprivation. Mammalian development occurs in a hypoxic environment and HIF-null mice therefore die in utero due to multiple embryonic and placental defects. Mouse embryonic stem cells do not differentiate into placental cells; therefore, trophoblast stem cells (TSCs) are used to study mouse placental development. Consistent with a requirement for HIF activity during placental development in utero, TSCs derived from HIF-null mice exhibit severe differentiation defects and fail to form trophoblast giant cells (TGCs) in vitro. Interestingly, differentiating TSCs induce HIF activity independent of oxygen tension via unclear mechanisms. Here, we show that altering the extracellular matrix (ECM) composition upon which TSCs are cultured changes their differentiation potential from TGCs to multinucleated syncytiotropholasts (SynTs) and blocks oxygen-independent HIF induction. We further find that modulation of Mitogen Activated Protein Kinase Kinase-1/2 (MAP2K1/2, MEK-1/2) signaling by ECM composition is responsible for this effect. In the absence of ECM-dependent cues, hypoxia-signaling pathways activate this MAPK cascade to drive HIF induction and redirect TSC fate along the TGC lineage. In addition, we show that integrity of the microtubule and actin cytoskeleton is critical for TGC fate determination. HIF-2α ensures TSC cytoskeletal integrity and promotes invasive TGC formation by interacting with c-MYC to induce non-canonical expression of Lim domain kinase 1-an enzyme that regulates microtubule and actin stability, as well as cell invasion. Thus, we find that HIF can integrate positional and metabolic cues from within the TSC niche to regulate placental development by modulating the cellular cytoskeleton via non-canonical gene expression.


Iatrogenic pseudoaneurysm of the superior gluteal artery presenting as pelvic mass with foot drop and sciatica: case report and review of literature.
We report an unusual case of a pseudoaneurysm of the superior gluteal artery as a complication of bone marrow biopsy. A 51-year-old man presented with sciatic pain and foot drop after undergoing bone marrow biopsy and was initially diagnosed as having degenerative disc disease based on his past medical history. Pelvic magnetic resonance imaging (MRI) revealed a large heterogeneous mass suggestive of a neurogenic tumor, but pulsatile blood was instead encountered during computed tomography (CT)-guided needle biopsy. Subsequent workup established the diagnosis of a superior gluteal artery pseudoaneurysm, which was treated with coil embolization, followed by surgical evacuation of the hematoma, which relieved his sciatic pain. However, the patient continues to have a persistent foot drop. Gluteal artery pseudoaneurysms are exceedingly uncommon but should be considered in the workup of a patient with gluteal pain or sciatic nerve palsy following trauma or medical procedures in the gluteal region.


Efficiently Training Vision Transformers on Structural MRI Scans for Alzheimer's Disease Detection.
Neuroimaging of large populations is valuable to identify factors that promote or resist brain disease, and to assist diagnosis, subtyping, and prognosis. Data-driven models such as convolutional neural networks (CNNs) have increasingly been applied to brain images to perform diagnostic and prognostic tasks by learning robust features. Vision transformers (ViT) - a new class of deep learning architectures - have emerged in recent years as an alternative to CNNs for several computer vision applications. Here we tested variants of the ViT architecture for a range of desired neuroimaging downstream tasks based on difficulty, in this case for sex and Alzheimer's disease (AD) classification based on 3D brain MRI. In our experiments, two vision transformer architecture variants achieved an AUC of 0.987 for sex and 0.892 for AD classification, respectively. We independently evaluated our models on data from two benchmark AD datasets. We achieved a performance boost of 5% and 9-10% upon fine-tuning vision transformer models pre-trained on synthetic (generated by a latent diffusion model) and real MRI scans, respectively. Our main contributions include testing the effects of different ViT training strategies including pre-training, data augmentation and learning rate warm-ups followed by annealing, as pertaining to the neuroimaging domain. These techniques are essential for training ViT-like models for neuroimaging applications where training data is usually limited. We also analyzed the effect of the amount of training data utilized on the test-time performance of the ViT via data-model scaling curves.Clinical Relevance- The models evaluated in this work could be trained on neuroimaging data to assist in diagnosis, subtyping and prognosis of Alzheimer's disease.


Ten things to get right for marine conservation planning in the Coral Triangle.
Systematic conservation planning increasingly underpins the conservation and management of marine and coastal ecosystems worldwide. Amongst other benefits, conservation planning provides transparency in decision-making, efficiency in the use of limited resources, the ability to minimise conflict between diverse objectives, and to guide strategic expansion of local actions to maximise their cumulative impact. The Coral Triangle has long been recognised as a global marine conservation priority, and has been the subject of huge investment in conservation during the last five years through the Coral Triangle Initiative on Coral Reefs, Fisheries and Food Security. Yet conservation planning has had relatively little influence in this region. To explore why this is the case, we identify and discuss 10 challenges that must be resolved if conservation planning is to effectively inform management actions in the Coral Triangle. These are: making conservation planning accessible; integrating with other planning processes; building local capacity for conservation planning; institutionalising conservation planning within governments; integrating plans across governance levels; planning across governance boundaries; planning for multiple tools and objectives; understanding limitations of data; developing better measures of progress and effectiveness; and making a long term commitment. Most important is a conceptual shift from conservation planning undertaken as a project, to planning undertaken as a process, with dedicated financial and human resources committed to long-term engagement.


Opportunities and Barriers to the Development and Use of Open Source Health Economic Models: A Survey.
Health economic (HE) models are routinely used to support health policy and resource allocation decisions but are often considered "black boxes" that may be prone to error and bias. Open source models (OSMs) have been advocated to increase the transparency, credibility, and reuse of HE models. Previous studies have demonstrated interest in OSMs among the health economics and outcomes research community, but the number of OSMs remains low. We conducted an online survey of ISPOR (the leading professional society for health economics and outcomes research) members' perspectives on the usefulness of OSMs and barriers to their development and implementation. Respondents (N = 230) included academics (27%), pharmaceutical (or related) industry representatives (23%), health research or consulting representatives (21%), governmental or nonprofit agency representatives (10%), and others (19%). Respondents were generally not familiar with barriers to the development and adoption of OSMs. Most agreed that OSMs would improve transparency (92%), efficiency (76%), and HE model reuse (86%) and promote confidence in using HE models (75%). The use of OSMs by health technology assessment authorities was considered a very important indicator of the usefulness of OSMs by 49% of respondents. Three-quarters of respondents perceived legal concerns and the ability to transfer data as important barriers to the development and use of OSMs. Respondents believe that OSMs could increase the transparency, efficiency, and credibility of HE models, but that several barriers hamper their widespread adoption. Our results suggest that fundamental changes may be needed across the health economics and outcomes research community if OSMs are to become widely adopted.


The effects of previous meniscus and anterior cruciate ligament injuries in patients with total knee arthroplasty.
Patients undergoing total knee replacements constitute a suitable population to study the natural history of traumatic joint injuries. We studied all the patients who received a TKA (Total knee arthroplasty) over the course of one year, in five different centers. The study included 474 patients who had undergone primary TKA for knee OA over a one-year period. In each patient, we analyzed age, sex, side of operation, weight, height and body mass index (BMI, kg/m2). BMI were stratified into four groups according to the WHO classification: normal (<25), overweight (>25 and <30), obese (>30 and <40), and morbidly obese (>40). In the TKA group, 74% of the patients were women, while in the THA group the percentage of men and women was similar. No differences were found in the sides operated on. Differences between both groups were found in knee alignment. Women were operated on more frequently for TKA, as there was a higher incidence of OA of the knee joints in women aged over 65. Patients who required a total knee arthroplasty are likely to have previously undergone surgery or trauma to the knee joints.


Implicit level set algorithms for modelling hydraulic fracture propagation.
Hydraulic fractures are tensile cracks that propagate in pre-stressed solid media due to the injection of a viscous fluid. Developing numerical schemes to model the propagation of these fractures is particularly challenging due to the degenerate, hypersingular nature of the coupled integro-partial differential equations. These equations typically involve a singular free boundary whose velocity can only be determined by evaluating a distinguished limit. This review paper describes a class of numerical schemes that have been developed to use the multiscale asymptotic behaviour typically encountered near the fracture boundary as multiple physical processes compete to determine the evolution of the fracture. The fundamental concepts of locating the free boundary using the tip asymptotics and imposing the tip asymptotic behaviour in a weak form are illustrated in two quite different formulations of the governing equations. These formulations are the displacement discontinuity boundary integral method and the extended finite-element method. Practical issues are also discussed, including new models for proppant transport able to capture 'tip screen-out'; efficient numerical schemes to solve the coupled nonlinear equations; and fast methods to solve resulting linear systems. Numerical examples are provided to illustrate the performance of the numerical schemes. We conclude the paper with open questions for further research. This article is part of the themed issue 'Energy and the subsurface'.


[Resistance of Brevipalpus phoenicis (Geijskes) (Acari: Tenuipalpidae) to acaricides that inhibit cellular respiration in citrus: cross-resistance and fitness cost].
Acaricides that inhibit cellular respiration play an important role in the control of Brevipalpus phoenicis (Geijskes) in citrus groves in Brazil. Studies were conducted to evaluate: (a) the variability in the susceptibility among B. phoenicis populations collected from citrus groves to cyhexatin, azocyclotin, propargite and sulphur; (b) cross-resistance relationships between propargite and azocyclotin, cyhexatin, dinocap, pyridaben and sulphur; and (c) the fitness cost associated with propargite resistance in B. phoenicis under laboratory conditions. A residual-type contact bioassay was used to characterize the susceptibility. The susceptibility was estimated with diagnostic concentrations based LC(95) of each acaricide. The cross-resistance was evaluated by characterizing the concentration-mortality responses of susceptible (S) and propargite-resistant (Propargite-R) strains. The fitness cost was evaluated by measuring the biological parameters of S and Propargite-R strains on citrus fruits at 25 +/- 1 degrees C and fotophase of 14h. Significant differences in the susceptibility of B. phoenicis were detected at diagnostic concentration of cyhexatin (survivorship from 16.3% to 80.5%), azocyclotin (from 3.0% to 15.0%), propargite (from 1.0% a 71.6%) and sulphur (from 9.0% to 82.6%). A low intensity of cross-resistance was detected between propargite and the acaricides azocyclotin (1.8-fold), cyhexatin (4.6-fold), dinocap (3.5-fold) and pyridaben (3.5-fold). On the other hand, the intensity of cross-resistance to sulphur (> 111-fold) was very high. There was no fitness cost associated with B. phoenicis resistance to propargite, based on biological parameters evaluated. Therefore, the use of these acaricides should also be done very carefully in resistance management of B. phoenicis to acaricides.


Population Dynamics of the Segregation Distorter Polymorphism of DROSOPHILA MELANOGASTER.
Two two-locus models of the population dynamics of the segregation distortion (SD) polymorphism of Drosophila melanogaster are described. One model is appropriate for understanding the population genetics of SD in nature, whereas the other is a special case appropriate for understanding an artificial population that has been extensively analysed. The models incorporate the general features of the Sd and Rsp loci which form the core of the SD system. It is shown that the SD polymorphism can be established only when there is sufficiently tight linkage between Sd and Rsp. An approximate treatment, valid for tight linkage, is given of all the equilibria of the system and their stabilities. It is shown that the observed composition of natural and artificial populations with respect to the Sd and Rsp loci is predicted well by the model, provided that restrictions are imposed on the fertilities of certain genotypes. Highly oscillatory paths towards equilibrium are usually to be expected on the basis of this model. The selection pressures on inversions introduced into this system are also investigated.


Effects of sodium replacement on uptake of the dipeptide glycylsarcosine by hamster jejunum in vitro.
We report observations on transport of the hydrolysis-resistant dipeptide glycylsarcosine by rings of everted hamster jejunum in vitro in the presence and absence of Na+, using several substituents for Na+: Li+, K+, Cs+, Tris, choline and mannitol. At most concentrations, mediated influx of glycylsarcosine was depressed by Li+, K+, Cs+ and Tris, though not abolished. At high concentrations, it was moderately increased by choline and mannitol. Under conditions in which the tissue could concentrate the peptide in the presence of Na+, uptake was greatly depressed by all the substituents and the ability to concentrate was abolished. The Kt of mediated influx was affected in different ways according to the substituent used. Kt was reduced by Li+ replacement of Na+ and increased by choline replacement. Vmax was greatly reduced by all metallic substituents but not by non-metallic substituents. Though the results cannot yet be satisfactorily interpreted, they suggest possible reasons for previous conflicting results and show that it is impossible to make the unqualified statement that transport of glycylsarcosine is 'Na+-dependent'. It is now doubtful whether transport of this peptide into the intestinal cells is by co-transport with Na+, and the whole matter of Na+-dependence of intestinal peptide transport is in question.


Gait and gait-related activities and fatigue in Parkinson's disease: what is the relationship?
To explore the relationship between gait and gait-related activities and fatigue in Parkinson's disease (PD). Twenty people with idiopathic PD (12 male, 8 female; mean age 64.6 +/- 7.96) and 10 age, sex and education matched controls (6 male, 4 female; mean age 63.5 +/- 7.03) wore an activity monitor for approximately 24 h, from which the percentage time walking and standing, the number of periods of walking greater than 10 sec were derived. Prior to monitoring, levels of fatigue (Multi-dimensional Fatigue Index, MFI) were evaluated. Activity data related to gait were compared for the waking hours of the day, reflected by the period between 6 am and 10 pm. Overall, PD subjects demonstrated reduced activity compared to controls, however there was no significant difference between PD and controls for the percentage time spent walking (p = 0.149), standing (p = 1.0) or engaged in periods of walking that were greater than 10 sec (p = 0.059). Significantly greater levels of fatigue were experienced by PD subjects in all MFI domains compared to controls (physical p = 0.001; activity p = 0.001; general p = 0.001; mental p = 0.028; motivation p = 0.024). There was no clear association between activity and fatigue in PD subjects. Evidence of increased fatigue is not supported by evidence of significantly decreased levels of activity in PD subjects. The relationship between activity and fatigue therefore remains unclear, highlighting the complex nature of the relationship between these factors. Further work is required to increase our understanding of the impact of fatigue on gait and gait-related activity so that improved management can be explored.


Involvement of Girdin in the determination of cell polarity during cell migration.
Cell migration is a critical cellular process that determines embryonic development and the progression of human diseases. Therefore, cell- or context-specific mechanisms by which multiple promigratory proteins differentially regulate cell migration must be analyzed in detail. Girdin (girders of actin filaments) (also termed GIV, Gα-interacting vesicle associated protein) is an actin-binding protein that regulates migration of various cells such as endothelial cells, smooth muscle cells, neuroblasts, and cancer cells. Here we show that Girdin regulates the establishment of cell polarity, the deregulation of which may result in the disruption of directional cell migration. We found that Girdin interacts with Par-3, a scaffolding protein that is a component of the Par protein complex that has an established role in determining cell polarity. RNA interference-mediated depletion of Girdin leads to impaired polarization of fibroblasts and mammary epithelial cells in a way similar to that observed in Par-3-depleted cells. Accordingly, the expression of Par-3 mutants unable to interact with Girdin abrogates cell polarization in fibroblasts. Further biochemical analysis suggests that Girdin is present in the Par protein complex that includes Par-3, Par-6, and atypical protein kinase C. Considering previous reports showing the role of Girdin in the directional migration of neuroblasts, network formation of endothelial cells, and cancer invasion, these data may provide a specific mechanism by which Girdin regulates cell movement in biological contexts that require directional cell movement.


Quantitative gated SPECT-derived phase analysis on gated myocardial perfusion SPECT detects left ventricular dyssynchrony and predicts response to cardiac resynchronization therapy.
The significance of left ventricular (LV) dyssynchrony for the prediction of response to cardiac resynchronization therapy (CRT) has been demonstrated. Parameters reflecting LV dyssynchrony (phase SD, histogram bandwidth) can be derived from gated myocardial perfusion SPECT (GMPS) using phase analysis. The feasibility of LV dyssynchrony assessment with phase analysis on GMPS using Quantitative Gated SPECT (QGS) software has not been demonstrated in patients undergoing CRT. The aim of the present study was to validate the QGS algorithm for phase analysis on GMPS in a direct comparison with echocardiography using tissue Doppler imaging (TDI) for LV dyssynchrony assessment. Also, prediction of response to CRT using GMPS and phase analysis was evaluated. Patients (n = 40) with severe heart failure (New York Heart Association class III-IV), an LV ejection fraction of no more than 35%, and a QRS complex greater than or equal to 120 ms were evaluated for LV dyssynchrony using GMPS and echocardiography with TDI. At baseline and after 6 mo of CRT, clinical status, LV volumes, and LV ejection fraction were evaluated. Patients with functional improvement were classified as CRT responders. Both histogram bandwidth (r = 0.69, r(2) = 0.48, SEE = 25.4, P < 0.01) and phase SD (r = 0.65, r(2) = 0.42, SEE = 26.8, P < 0.01) derived from GMPS correlated significantly with TDI for assessment of LV dyssynchrony. At baseline, CRT responders showed a significantly larger histogram bandwidth (94 degrees +/- 23 degrees vs. 68 degrees +/- 21 degrees , P < 0.01) and a larger phase SD (26 degrees +/- 6 degrees vs. 18 degrees +/- 5 degrees , P < 0.01) than did nonresponders. Receiver-operating-characteristic curve analysis identified an optimal cutoff value of 72.5 degrees for histogram bandwidth to predict CRT response, yielding a sensitivity of 83% and a specificity of 81%. For phase SD, sensitivity and specificity similar to those for histogram bandwidth were obtained at a cutoff value of 19.6 degrees . QGS phase analysis on GMPS correlated significantly with TDI for the assessment of LV dyssynchrony. Moreover, a high accuracy for prediction of response to CRT was obtained using either histogram bandwidth or phase SD.


Sources of distortion in functional MRI data.
Functional magnetic resonance image (fMRI) experiments rely on the ability to detect subtle signal changes in magnetic resonance image time series. Any areas of signal change that correlate with the neurological stimulus can then be identified and compared with a corresponding high-resolution anatomical scan. This report reviews some of the several artefacts that are frequently present in fMRI data, degrading their quality and hence their interpretation. In particular, the effects of magnetic field inhomogeneities are described, both on echo planar imaging (EPI) data and on spiral imaging data. The modulation of these distortions as the subject moves in the magnet is described. The effects of gradient coil nonlinearities and EPI ghost correction schemes are also discussed.


Stimulation of endocannabinoid formation in brain slice cultures through activation of group I metabotropic glutamate receptors.
Activation of group I metabotropic glutamate (mGlu) receptors drives the endocannabinoid system to cause both short- and long-term changes of synaptic strength in the striatum, hippocampus, and other brain areas. Although there is strong electrophysiological evidence for a role of endocannabinoid release in mGlu receptor-dependent plasticity, the identity of the endocannabinoid transmitter mediating this phenomenon remains undefined. In this study, we show that activation of group I mGlu receptors triggers the biosynthesis of the endocannabinoid 2-arachidonoylglycerol (2-AG), but not anandamide, in primary cultures of corticostriatal and hippocampal slices prepared from early postnatal rat brain. Pharmacological studies suggest that 2-AG biosynthesis is initiated by activation of mGlu5 receptors, is catalyzed by phospholipase C (PLC) and 1,2-diacylglycerol lipase (DGL) activities, and is dependent on intracellular Ca2+ ions. Realtime polymerase chain reaction and immunostaining analyses indicate that DGL-beta is the predominant DGL isoform expressed in corticostriatal and hippocampal slices and that this enzyme is highly expressed in striatal neurons, where it is colocalized with PLC-beta1. The results suggest that 2-AG is a primary endocannabinoid mediator of mGlu receptor-dependent neuronal plasticity.


Survival trends in mantle cell lymphoma in the United States over 16 years 1992-2007.
A retrospective analysis was done using the Surveillance, Epidemiology, and End-Results (SEER) database to determine the trends in overall survival and identify prognostic factors in patients with mantle cell lymphoma (MCL). In total 5367 cases of MCL identified from 1992 to 2007 were split into three cohorts, group 1(1992-1999), group 2 (2000-2003) and group 3 (2004-2007). Survival was analyzed using the Cox proportional hazards model to correct for age, gender and stage of disease. The proportion of patients with advanced disease at diagnosis, male gender and advanced age increased over time and these were all associated with increased mortality. The overall survival remained unchanged. However, when adjusted for the increased proportion of patients with poor prognostic features noted above, we found a significant improvement in survival. The adjusted model also showed an improvement in predicted survival over time in patients with advanced stage. No change in survival was seen in patients with localized disease. Although this analysis is not designed to evaluate specific treatment modalities, these data suggest that the development of new treatment strategies over the past decade may be impacting the survival of patients with advanced MCL despite the finding that the overall survival remains unchanged in the general US population.


Cervical Cancer Prevention Through HPV Vaccination in Low- and Middle-Income Countries in Asia
Cervical cancer is ranked the first or second most common cancer in women of low- and middle-income countries (LMICs) in Asia. Cervical cancer is almost exclusively caused by human papillomavirus (HPV), and majority of the cases can be prevented with the use of HPV vaccines. The HPV vaccines have demonstrated high vaccine efficacies against HPV infection and cervical cancer precursors in clinical and post-marketing studies, and are in use in most high-income countries. However, their use in LMICs are limited mainly due to the high costs and logistics in delivering multiple doses of the vaccine. Other issues such as the safety of the vaccines, social and cultural factors, as well as poor knowledge and awareness of the virus have also contributed to the low uptake of the vaccine. This mini-review focuses on the need for HPV vaccine implementation in Asia given the substantial disease burden and underuse of HPV vaccines in LMICs in this region. In addition, the progress towards HPV vaccine introduction, and barriers preventing further rollout of these essential, life-saving vaccines are also discussed in this article.


A compensatory mutation provides resistance to disparate HIV fusion inhibitor peptides and enhances membrane fusion.
Fusion inhibitors are a class of antiretroviral drugs used to prevent entry of HIV into host cells. Many of the fusion inhibitors being developed, including the drug enfuvirtide, are peptides designed to competitively inhibit the viral fusion protein gp41. With the emergence of drug resistance, there is an increased need for effective and unique alternatives within this class of antivirals. One such alternative is a class of cyclic, cationic, antimicrobial peptides known as θ-defensins, which are produced by many non-human primates and exhibit broad-spectrum antiviral and antibacterial activity. Currently, the θ-defensin analog RC-101 is being developed as a microbicide due to its specific antiviral activity, lack of toxicity to cells and tissues, and safety in animals. Understanding potential RC-101 resistance, and how resistance to other fusion inhibitors affects RC-101 susceptibility, is critical for future development. In previous studies, we identified a mutant, R5-tropic virus that had evolved partial resistance to RC-101 during in vitro selection. Here, we report that a secondary mutation in gp41 was found to restore replicative fitness, membrane fusion, and the rate of viral entry, which were compromised by an initial mutation providing partial RC-101 resistance. Interestingly, we show that RC-101 is effective against two enfuvirtide-resistant mutants, demonstrating the clinical importance of RC-101 as a unique fusion inhibitor. These findings both expand our understanding of HIV drug-resistance to diverse peptide fusion inhibitors and emphasize the significance of compensatory gp41 mutations.


Clinical, cerebrospinal fluid, and neuroimaging findings in COVID-19 encephalopathy: a case series.
To describe the clinical, neurological, neuroimaging, and cerebrospinal fluid (CSF) findings associated with encephalopathy in patients admitted to a COVID-19 tertiary reference center. We retrospectively reviewed records of consecutive patients with COVID-19 evaluated by a consulting neurology team from March 30, 2020 through May 15, 2020. Fifty-five patients with confirmed SARS-CoV-2 were included, 43 of whom showed encephalopathy, and were further divided into mild, moderate, and severe encephalopathy groups. Nineteen patients (44%) had undergone mechanical ventilation and received intravenous sedatives. Eleven (26%) patients were on dialysis. Laboratory markers of COVID-19 severity were very common in encephalopathy patients, but did not correlate with the severity of encephalopathy. Thirty-nine patients underwent neuroimaging studies, which showed mostly non-specific changes. One patient showed lesions possibly related to CNS demyelination. Four had suffered an acute stroke. SARS-CoV-2 was detected by RT-PCR in only one of 21 CSF samples. Two CSF samples showed elevated white blood cell count and all were negative for oligoclonal bands. In our case series, the severity of encephalopathy correlated with higher probability of death during hospitalization (OR = 5.5 for each increment in the degree of encephalopathy, from absent (0) to mild (1), moderate (2), or severe (3), p < 0.001). In our consecutive series with 43 encephalopathy cases, neuroimaging and CSF analysis did not support the role of direct viral CNS invasion or CNS inflammation as the cause of encephalopathy.


Effect of statin therapy on muscle symptoms: an individual participant data meta-analysis of large-scale, randomised, double-blind trials.
Statin therapy is effective for the prevention of atherosclerotic cardiovascular disease and is widely prescribed, but there are persisting concerns that statin therapy might frequently cause muscle pain or weakness. We aimed to address these through an individual participant data meta-analysis of all recorded adverse muscle events in large, long-term, randomised, double-blind trials of statin therapy. Randomised trials of statin therapy were eligible if they aimed to recruit at least 1000 participants with a scheduled treatment duration of at least 2 years, and involved a double-blind comparison of statin versus placebo or of a more intensive versus a less intensive statin regimen. We analysed individual participant data from 19 double-blind trials of statin versus placebo (n=123 940) and four double-blind trials of a more intensive versus a less intensive statin regimen (n=30 724). Standard inverse-variance-weighted meta-analyses of the effects on muscle outcomes were conducted according to a prespecified protocol. Among 19 placebo-controlled trials (mean age 63 years [SD 8], with 34 533 [27·9%] women, 59 610 [48·1%] participants with previous vascular disease, and 22 925 [18·5%] participants with diabetes), during a weighted average median follow-up of 4·3 years, 16 835 (27·1%) allocated statin versus 16 446 (26·6%) allocated placebo reported muscle pain or weakness (rate ratio [RR] 1·03; 95% CI 1·01-1·06). During year 1, statin therapy produced a 7% relative increase in muscle pain or weakness (1·07; 1·04-1·10), corresponding to an absolute excess rate of 11 (6-16) events per 1000 person-years, which indicates that only one in 15 ([1·07-1·00]/1·07) of these muscle-related reports by participants allocated to statin therapy were actually due to the statin. After year 1, there was no significant excess in first reports of muscle pain or weakness (0·99; 0·96-1·02). For all years combined, more intensive statin regimens (ie, 40-80 mg atorvastatin or 20-40 mg rosuvastatin once per day) yielded a higher RR than less intensive or moderate-intensity regimens (1·08 [1·04-1·13] vs 1·03 [1·00-1·05]) compared with placebo, and a small excess was present (1·05 [0·99-1·12]) for more intensive regimens after year 1. There was no clear evidence that the RR differed for different statins, or in different clinical circumstances. Statin therapy yielded a small, clinically insignificant increase in median creatine kinase values of approximately 0·02 times the upper limit of normal. Statin therapy caused a small excess of mostly mild muscle pain. Most (>90%) of all reports of muscle symptoms by participants allocated statin therapy were not due to the statin. The small risks of muscle symptoms are much lower than the known cardiovascular benefits. There is a need to review the clinical management of muscle symptoms in patients taking a statin. British Heart Foundation, Medical Research Council, Australian National Health and Medical Research Council.


EU member state government contribution to alternative methods.
Article 47 of the new EU Directive 2010/63/EU on the protection of animals used for scientific purposes requires national governments to contribute to the development and promotion of alternative methods. A recent survey of EU member states found that reported funding of alternative (3Rs) methods totalled € 18.7 million in 2013, provided by only seven countries (Austria, Belgium, Denmark, Finland, Germany, Sweden, and the UK). There were indications that the contributions of some of these countries have increased since the implementation of the new Directive. However, funding of alternatives is between 0 and 0.036% of national science R&D expenditure and nearly half of the countries that responded reported that they do not specifically contribute. Data (and, by assumption, financial contribution) remains unavailable from half of the member states across the EU, regardless of the method of collection.


Expression of the second lysine decarboxylase gene of Escherichia coli.
Certain amino acids are substrates for two decarboxylase enzymes in Escherichia coli, one inducible by anaerobic growth at low pH and the other constitutive. In the case of lysine, an inducible decarboxylase (CadA) has been extensively characterized, but evidence for the existence of a second lysine decarboxylase is fragmentary and uncertain. This paper confirms that a second lysine decarboxylase is encoded by a locus (ldc) previously suggested to be a lysine decarboxylase gene on the basis of sequence comparisons. Overexpression of the cloned gene provided sufficient quantities of enzyme in cell-free extracts for preliminary examination of the properties of the ldc gene product, Ldc. The enzyme is active over a broad range of pH with an optimum at 7.6, much higher than that of CadA, about 5.5. The temperature optimum for both enzymes is similar, at about 52 degrees C, but Ldc is more readily inactivated by heat than CadA. Expression of ldc from its own promoter was very weak for cells growing in a variety of media, although a low level of lysine decarboxylase was present in cells that carried the ldc region on an oligo-copy plasmid when these were grown in minimal-glucose medium. Northern analysis of RNA extracted from such cells revealed a transcript whose length corresponded to that of the ldc gene, suggesting that ldc is normally transcribed from a promoter immediately upstream. However, most of the ldc mRNA was shorter, indicating degradation or premature termination. The ldc upstream sequence promoted transcription of a lacZ gene to which it was fused. Introduction of the upstream sequence as an insert in a multicopy vector increased transcription of the resident lacZ fusion. The low level of expression in single copy, the emergence of expression when the gene is present at moderate copy number, and the derepression by the upstream sequence in trans imply that this second lysine decarboxylase gene may not be constitutive but subject to specific repression by a factor which remains to be identified.


Bi-ventricular pacing improves pump function only with adequate myocardial perfusion in canine hearts with pseudo-left bundle branch block.
Bi-ventricular (BiV) pacing is an effective therapy for the treatment of cardiac electromechanical (EM) dysfunction. The reason(s), however, for therapy non-response in approximately one-third of the subjects remains unclear, especially as it relates to myocardial perfusion and pacing location. In this study, we examined how acute BiV pacing response may be related to underlying myocardial perfusion coupled with pacing near or distant to the area of perfusion. In 10 open-chest anesthetized canines, coronary blood flow to the left ventricular (LV) anterior wall (AW: n = 5) and lateral wall (LW: n = 5) was controlled during four pacing conditions: right atrial, right ventricular (pseudo-left bundle branch block; [pseudo-LBBB]), BiV-LW and BiV-AW. Local EM function (piezo-electrical crystals and electrodes), along with global hemodynamic parameters, were measured during all pacing conditions at three coronary perfusion rates (≥0.40 mL/min/g, 0.20-0.40 mL/min/g and <0.20 mL/min/g). A positive BiV therapy response was assessed by a significant increase in the maximum cardiac output compared with the pseudo-LBBB condition. Despite no improvement in QRS duration, BiV-LW pacing improved LV function compared with the pseudo-LBBB pacing condition (P value <0.01). This improvement with BiV-LW pacing was seen above a certain myocardial perfusion threshold and was independent of any increases in regional coronary blood flow with BiV pacing. At lower myocardial perfusion rates, LV function was not improved with BiV pacing at any location. This study underscores the significance of even mild ischemia on BiV pacing response.


Biological activity and coordination modes of copper(II) complexes of Schiff base-derived coumarin ligands.
The coordination modes of copper(II) complexes of Schiff base-derived coumarin ligands, which had previously shown good anti-Candida activity, were investigated by pH-potentiometric and UV-Vis spectroscopic methods. These studies confirmed the coordination mode of the ligands to be through the N of the imine and deprotonated phenol of the coumarin-derived ligand in solution. In addition, the more active complexes and their corresponding ligands were investigated in the presence of copper(II) in liquid and frozen solution by ESR spectroscopic methods. A series of secondary amine derivatives of the Schiff base ligands, were isolated with good solubility characteristics but showed little anti-Candida activity. However, cytotoxicity studies of the secondary amines, together with the copper complexes and their corresponding ligands, against human colon cancer and human breast cancer cells identified the chemotherapeutic potential of these new ligands.


Energy intake and meal portions: associations with BMI percentile in U.S. children.
We examined relationships of eating patterns and reported energy intake (rEI) with BMI percentile in U.S. children. Two 24-hour dietary recalls from the Continuing Surveys of Food Intakes by Individuals 1994 to 1996 and 1998 (1005 boys, 990 girls) were averaged, and children were categorized into three age groups: 3 to 5 years (n = 1077), 6 to 11 years (n = 537), and 12 to 19 years (n = 381). Physiologically implausible reports due to reporting bias or abnormal intake (rEI outside +/-18% to 23% of predicted energy requirements; pER) were identified. rEI averaged 109 +/- 34% and 100 +/- 10% of pER in the total and plausible samples, respectively. EI was overreported more in younger children and underreported more in overweight older children. Children with plausible rEI (45.3% of sample) averaged 4.7 eating occasions/d, 589 kcal/meal, 223 kcal/snack, and 2038 kcal/d. rEI was not associated with BMI percentile in the total sample. In the plausible sample, rEI, meal portion size, and meal energy were positively associated with BMI percentile in boys 6 to 11 years and in children 12 to 19 years. No relationships were found in children 3 to 5 years and girls 6 to 11 years. Relationships were more consistent and stronger in the plausible compared with the total sample. Excluding implausible dietary reports may be necessary for discerning dietary associations with BMI percentile. EI and meal, but not snack, patterns may play a quantitatively greater role in weight regulation as children age.


Adolescents and young adults with Down syndrome presenting to a medical clinic with depression: co-morbid obstructive sleep apnea.
Adolescents and young adults with Down syndrome (DS) sometimes experience new-onset mood disorder and decline in adaptive skills. The clinical phenomenon is poorly characterized and its pathogenesis is not understood. The possible contribution of obstructive sleep apnea syndrome (OSAS) to this phenomenon has not been studied. Subjects were ascertained as a convenience sample through our clinic for persons with DS and medical or mental health concerns between 2004 and 2009. When mood symptoms were present an axis I diagnosis was made using DSM-IV-R criteria. Subjects without an axis I diagnosis served as controls. The Reiss scales for children's dual diagnosis and the aberrant behavior checklist (ABC) were completed by caretakers. Twenty-eight cases meeting criteria for major depressive episode (MDE) and nine controls without psychopathology were referred for overnight polysomnography (PSG). Functional decline was reported in 19 (68%) of cases with MDE, but none of the controls. Twenty-four (86%) cases had OSAS compared with only 4 (44%) of controls. Moderate-severe OSAS was present in 15 (54%) of cases compared to only 1 (11%) of controls. Intermittent sleep-associated hypoxia and REM sleep deficits were also more frequent in cases. Across all subjects, prior tonsillectomy was not related to the presence or absence of OSAS. Our findings suggest that OSAS may be a common co-morbidity in adolescents and younger adults with DS and depression. Recognition of this association maybe critical to understanding the pathogenesis and management of mood-related disorders, and functional decline in affected individuals.


Cathepsin B-Activated Fluorescent and Photoacoustic Imaging of Tumor.
Early diagnosis is crucial to the treatment of cancer. Cathepsin B (CTB) plays an important role in numerous cancers, which is a promising biomarker for early diagnosis of cancer. It is necessary to exploit new probes for visualization of CTB in vivo. Fluorescent/photoacoustic (FL/PA) imaging is a powerful tool for in vivo study which possesses both excellent sensitivity and spatial resolution. To our knowledge, there has been no FL/PA probe to image CTB in vitro or in vivo. Therefore, we developed two CTB-activated FL/PA probes HCy-Cit-Val and HCy-Gly-Leu-Phe-Gly, which could successfully monitor CTB activity in vivo. Both two probes had excellent sensitivity and selectivity in vitro. Cell imaging showed that HCy-Cit-Val or HCy-Gly-Leu-Phe-Gly could image endogenous CTB in lysosome with 6.8-fold or 5.1-fold enhancement of the FL signal and 5.8-fold or 3.4-fold enhancement of the PA signal compared to their inhibitor contrast groups. Tumor imaging in vivo further confirmed the good applicability of these two probes to monitor CTB activity with high sensitivity and spatial resolution. Moreover, the property of HCy-Cit-Val is superior to HCy-Gly-Leu-Phe-Gly due to the higher catalytic efficiency of CTB toward HCy-Cit-Val than HCy-Gly-Leu-Phe-Gly. We envision that our FL/PA probe HCy-Cit-Val will be suitable for clinical early diagnosis of CTB-related cancer in the near future.


Asymmetries in attention as revealed by fixations and saccades.
Neurologically normal individuals devote more attention to the left side; an asymmetry known as pseudoneglect, which reflects right hemisphere involvement in visuospatial attention. The role of eye movements in attentional asymmetries has received little consideration, particularly in terms of the greyscales task. Stimulus length, elevation, and presentation duration were manipulated, while monitoring eye movements during the greyscales task. Region of interest analyses compared time spent examining each quadrant of the stimulus. Further, saccades were examined in conjunction with fixations to gain an understanding of overall eye movement patterns. Scatterplots combining x-and y-coordinates illustrate mean eye position. Results demonstrated a comparison strategy was used, where the dark portions of each rectangle were fixated. Mean eye position was within the lower left quadrant. The left visual field was inspected most for the baseline condition. Interestingly, the lower visual field was examined most when duration, length, or elevation was manipulated. Eye movement patterns provide a possible explanation for why correlations are y not observed between visuospatial tasks. Different strategies, based on specific-task demands, are likely to be used, which in turn, engage separate aspects of visuospatial attention.


Constraints on melt movement beneath the East Pacific Rise from 230Th-238U disequilibrium.
We report 230Th-238U disequilibrium data on mid-ocean ridge basalts recovered 5 to 40 kilometers off the ridge axis near 9 degrees 30'N of the East Pacific Rise. These data indicate near-symmetrical eruptions of normal mid-ocean ridge basalts (NMORBs) and incompatible element-enriched mid-ocean ridge basalts (EMORBs) as far as 20 kilometers off axis. Our results suggest large-scale subsurface lateral transport of NMORB melt at 19 to 21 centimeters per year and also provide constraints on the petrogenesis of EMORBs of off-axis origin.


Initiation of osteoporosis treatment after bone mineral density testing.
The aim of the study was to describe initiation of osteoporosis drug therapy after bone mineral density (BMD) testing and to determine any association with BMD test results obtained, physician factors, or both. The setting was the Kaiser Foundation Health Plan (KFHP), a large health maintenance organization (HMO) in Northern California. Data were collected from bone densitometry centers at four KFHP medical centers sites in Sacramento, San Rafael, Fresno, and Oakland. We identified 17,290 women aged > or = 45 years who had BMD testing between January 1, 1997 and June 30, 1999. After excluding those for whom any osteoporosis drugs were prescribed in the year before testing, 8020 women were available for analysis. Using logistic regression, we examined the association between BMD diagnosis (i.e., osteoporosis or osteopenia versus normal) and initiation of drug therapy, for osteoporosis (including hormone replacement therapy (HRT), alendronate, etidronate, raloxifene and calcitonin) within 6 months after the test. Among the 8020 women, 1934 (24%) filled a prescription for an osteoporosis drug within 6 months after BMD testing. Compared with women who had a normal BMD test result, women diagnosed with osteopenia were nearly 4 times more likely (OR = 3.7; CI = 3.0-4.4), and women diagnosed with osteoporosis were 15 times more likely (OR = 15.0; CI = 12.5-18.1), to fill a prescription for an osteoporosis drug within 6 months after BMD testing. Women with high exposure to corticosteroid agents were twice as likely (OR = 2.1; CI = 1.7-2.7) to start osteoporosis drug therapy compared with women who were not similarly exposed; women diagnosed with recent osteoporotic fractures were 50% more likely (OR = 1.5; CI = 1.2-1.9) to begin therapy than women without such fractures. Despite the strong association between BMD and initiating treatment, nearly half the osteoporotic women did not initiate treatment. In addition, we found that age strongly influenced choice of osteoporotic drug. Compared with osteoporotic women aged 45-54 years, women aged 55-64 years who started drug therapy were 40% more likely (OR = 1.4; CI = 1.0-2.2) and women aged > or = 65 years were twice as likely (OR = 2.0; CI = 1.4-2.8) to start non-HRT drugs. BMD test results indicating osteoporosis were thus strongly associated with increased likelihood of beginning drug therapy, and half of such women initiated therapy. Drug initiation was also associated with other factors, including age, use of corticosteroid agents, recent fracture, and physician characteristics. However, these factors showed much weaker associations than those found for BMD. Health care providers must consider whether test results will influence treatment decisions, and our data indicate that results of BMD testing do influence management decisions regarding osteoporosis drug use for women.


Breast cancer among Hispanics, American Indians and non-Hispanic whites in New Mexico.
Variation in breast cancer occurrence among women in New Mexico's three major ethnic groups has not previously been assessed. The address the descriptive epidemiology of breast cancer in New Mexico Hispanics, American Indians, and non-Hispanic whites, we calculated incidence rates from population-based registry data covering 1969-1987 and mortality data collected from 1958 to 1987. Breast cancer incidence and mortality rates for New Mexico's non-Hispanic white women were comparable to those for white women nationwide. In contrast, American Indian women had extremely low incidence and mortality rates for breast cancer; rates for Hispanics were intermediate, but well below those for non-Hispanic white women throughout the study period. Pronounced temporal trends in breast cancer occurrence were evident among Hispanic women, with the incidence rate increasing by 56% over the 19 years of available data and the mortality rate increasing by nearly 100% over 30 years. Age-specific incidence and mortality rates increased at all ages for successive birth cohorts of Hispanic women. For non-Hispanic whites, increasing incidence and mortality rates were also observed, but the increments were much smaller, approximately 15% for incidence and 30% for mortality. Our data show substantial ethnic differences in breast cancer incidence and mortality in New Mexico, suggesting the need for aetiological investigations to assist in controlling this disease.


Emergence of epidemic Neisseria meningitidis serogroup X meningitis in Togo and Burkina Faso.
Serogroup X meningococci (NmX) historically have caused sporadic and clustered meningitis cases in sub-Saharan Africa. To study recent NmX epidemiology, we analyzed data from population-based, sentinel and passive surveillance, and outbreak investigations of bacterial meningitis in Togo and Burkina Faso during 2006-2010. Cerebrospinal fluid specimens were analyzed by PCR. In Togo during 2006-2009, NmX accounted for 16% of the 702 confirmed bacterial meningitis cases. Kozah district experienced an NmX outbreak in March 2007 with an NmX seasonal cumulative incidence of 33/100,000. In Burkina Faso during 2007-2010, NmX accounted for 7% of the 778 confirmed bacterial meningitis cases, with an increase from 2009 to 2010 (4% to 35% of all confirmed cases, respectively). In 2010, NmX epidemics occurred in northern and central regions of Burkina Faso; the highest district cumulative incidence of NmX was estimated as 130/100,000 during March-April. Although limited to a few districts, we have documented NmX meningitis epidemics occurring with a seasonal incidence previously only reported in the meningitis belt for NmW135 and NmA, which argues for development of an NmX vaccine.


Different excitation-contraction coupling mechanisms exist in squid, cuttlefish and octopod mantle muscle
Excitation-contraction (EC) coupling was studied in central zone mantle muscle fibres of a squid (Alloteuthis subulata), a cuttlefish (Sepia officinalis) and an octopod (Eledone cirrhosa). Thin slices of muscle were used for twitch experiments and enzymatic isolation of single fibres for whole-cell patch-clamp studies. The current required for a supramaximal twitch response during direct stimulation of muscle slices was lower for squid than for cuttlefish. In squid, but not in cuttlefish, the current-response relationship was independent of slice thickness (range 0.1-0.5 mm). Twitches of squid and cuttlefish slices were reversibly abolished by removal of extracellular Ca2+. In squid, but not in cuttlefish, the current-response relationship was Na+-dependent, and in the absence of Na+ higher current strengths were required to generate a supramaximal response. In whole-cell voltage-clamp experiments on isolated muscle fibres from squid, cuttlefish and Eledone cirrhosa, a sustained inward current was recorded upon depolarisation. This current was blocked by 5 mmol l-1 Co2+ and suppressed by 10 micromol l-1 nifedipine. In squid, an additional inward fast-activating transient current was seen which was blocked by 2 micromol l-1 tetrodotoxin and depolarised holding potentials. The fast current represents a voltage-activated Na+ channel, and the slow currents represent L-type Ca2+ channels. We conclude that squid possess a specialised rapid EC coupling mechanism in central zone fibres that is absent in cuttlefish and Eledone cirrhosa.


Temporal and quantitative correlations between insulin secretion and stably elevated or oscillatory cytoplasmic Ca2+ in mouse pancreatic beta-cells.
An increase in cytoplasmic Ca2+ in beta-cells is a key step in glucose-induced insulin secretion. However, whether changes in cytoplasmic free Ca2+ ([Ca2+]i) directly regulate secretion remains disputed. This question was addressed by investigating the temporal and quantitative relationships between [Ca2+]i and insulin secretion. Both events were measured simultaneously in single mouse islets loaded with fura-PE3 and perifused with a medium containing diazoxide (to prevent any effect of glucose on the membrane potential) and either 4.8 or 30 mmol/l K+. Continuous depolarization with 30 mmol/l K+ in the presence of 15 mmol/l glucose induced a sustained rise in [Ca2+]i and insulin release. No oscillations of secretion were detected even after mathematical analysis of the data (pulse, spectral and sample distribution analysis). In contrast, alternating between 30 and 4.8 mmol/l K+ (1 min/2 min or 2.5 min/5 min) triggered synchronous [Ca2+]i and insulin oscillations of regular amplitude in each islet. A good correlation was found between [Ca2+]i and insulin secretion, and it was independent of the presence or absence of oscillations. This quantitative correlation between [Ca2+]i and insulin secretion was confirmed by experiments in which extracellular Ca2+ was increased or decreased (0.1-2.5 mmol/l) stepwise in the presence of 30 mmol/l K+. This resulted in parallel stepwise increases or decreases in [Ca2+]i and insulin secretion. However, while the successive [Ca2+]i levels were unaffected by glucose, each plateau of secretion was much higher in 20 than in 3 mmol/l glucose. In conclusion, in our preparation of normal mouse islets, insulin secretion oscillates only when [Ca2+]i oscillates in beta-cells. This close temporal relationship between insulin secretion and [Ca2+]i changes attests of the regulatory role of Ca2+. There also exists a quantitative relationship that is markedly influenced by the concentration of glucose.


Soliton propagation and soliton collision in double-doped fibers with a non-Kerr-like nonlinear refractive-index change.
A physical mechanism for a non-Kerr-like nonlinear refractive-index change in double-doped fibers is proposed, which shows for realistic material parameters a bistable (or two-valued) soliton regime with a large bistability range. The collision of two bistable solitons with a frequency difference in such materials is investigated. Solitons of the lower solution branch retain their shape after the collision. In contrast, two solitons of the upper solution branch with a low-frequency difference fuse to a single high-energy soliton after the collision. The possible applications of this effect for nonlinear photonic switching are discussed.


Late iliac artery aneurysms and occlusive disease after aortic tube grafts for abdominal aortic aneurysm repair. A 35-year experience.
Controversy continues over whether patients treated with straight Dacron aortic tube grafts for an abdominal aortic aneurysm remain at significant risk for subsequent development of iliac aneurysm or occlusive disease. To address this issue, the authors performed a population-based analysis of 432 patients who had an abdominal aortic aneurysm diagnosed between 1951 and 1984. Aneurysm repair was performed eventually in 206 patients (48%). To ascertain differences in late development of graft-related complications, iliac aneurysms, and arterial occlusions, the authors compared all tube-graft patients with similar numbers of bifurcated-graft patients matched for age and year of operation. In the tube-graft group, no subsequent clinically evident or autopsy-proven iliac aneurysms or iliac occlusive disease were noted. Over a mean follow-up of 6 years (range, 4 to 18 years), new aortic aneurysms occurred in the proximal aorta in both tube and bifurcated-graft patients (5.0% and 2.5%, respectively). In contrast the cumulative incidence of graft-related complications was higher with a bifurcated prosthesis (12.8%) compared with a straight graft (5.0%) (p = 0.15). These problems generally occurred 5 to 15 years postoperatively and emphasize the need for long-term graft surveillance. The authors conclude that straight tube-grafts for repair of abdominal aortic aneurysms provide excellent late patency with minimal risk of subsequent iliac aneurysm development.


ZD6474, a vascular endothelial growth factor receptor tyrosine kinase inhibitor with additional activity against epidermal growth factor receptor tyrosine kinase, inhibits orthotopic growth and angiogenesis of gastric cancer.
Vascular endothelial growth factor (VEGF) and epidermal growth factor (EGF) have been strongly implicated in the growth and metastasis of gastric cancer. The purpose of this study was to examine the effects of ZD6474, an inhibitor of inhibitor of VEGF receptor (VEGFR) tyrosine kinase with additional activity against EGF receptor (EGFR), on tumor growth and angiogenesis in an orthotopic model of gastric cancer. In vitro, ZD6474 inhibited human umbilical vascular endothelial cell and TMK-1 human gastric tumor cell proliferation in a dose-dependent fashion. EGF-mediated activation of EGFR and Erk-1/2 was decreased in tumor cells after ZD6474 treatment. In addition, VEGF-mediated activation of VEGFR2 and Erk-1/2 was decreased in human umbilical vascular endothelial cells. TMK-1 human gastric adenocarcinoma cells were injected into the gastric wall of nude mice. ZD6474 therapy was initiated on day 10. Mice (n = 14 per group) were treated p.o. with (a) 1% Tween 80 (control), (b) 50 mg/kg/d ZD6474, or (c) 100 mg/kg/d ZD6474. Mice were sacrificed on day 33. Tumors from each group were stained for markers of blood vessels, pericytes, proliferation, and apoptosis. ZD6474 at both 50 and 100 mg/kg/d led to marked inhibition of tumor growth (P < 0.05). ZD6474 reduced tumor cell proliferation by 48% in the 50 mg/kg/d group and 65% in the 100 mg/kg/d group (P < 0.03) and increased tumor cell apoptosis (P < 0.001) in vivo. ZD6474 led to a 69% decrease in microvessel density in the 50 mg/kg/d group (P < 0.001) and a 62% decrease in the 100 mg/kg/d group (P < 0.001). Although microvessel density was decreased by ZD6474, the remaining vessels showed a relatively higher percentage of pericyte coverage (3-fold increase; P < 0.001), perhaps reflecting selective loss of uncovered vessels in the ZD6474 group. In conclusion, therapies such as ZD6474 that target two distinct aspects of tumor growth, angiogenesis and tumor cell proliferation, warrant further investigation.


Italian Cohort of the Nivolumab EAP in Squamous NSCLC: Efficacy and Safety in Patients With CNS Metastases.
Brain metastases are an additional challenge in patients with non-small-cell lung cancer (NSCLC) because most chemotherapy agents cannot cross the blood-brain barrier. Nivolumab has demonstrated efficacy in patients with advanced squamous NSCLC, but because patients with central nervous system (CNS) metastases are typically excluded from registration trials, 'field-practice' data are needed. Patients in the Italian cohort of the Expanded Access Program (EAP) who had CNS metastases at baseline were analyzed. Thirty-seven patients with CNS metastases received a median of six doses of nivolumab. Three patients (8%) had grade 3-4 adverse events and one patient discontinued due to an adverse event. The objective response rate was 19%. Median overall survival was 5.8 (95% confidence interval=1.9-9.8) months and median progression-free survival was 4.9 (95% confidence interval=2.7-7.1) months. The safety and efficacy of nivolumab in patients with CNS metastases appear to be similar to those seen in the overall EAP cohort in Italy.


An unmodified heptadecapeptide pheromone induces competence for genetic transformation in Streptococcus pneumoniae.
Competence for genetic transformation in Streptococcus pneumoniae has been known for three decades to arise in growing cultures at a critical cell density, in response to a secreted protease-sensitive signal. We show that strain CP1200 produces a 17-residue peptide that induces cells of the species to develop competence. The sequence of the peptide was found to be H-Glu-Met-Arg-Leu-Ser-Lys-Phe-Phe-Arg-Asp-Phe-Ile-Leu-Gln-Arg- Lys-Lys-OH. A synthetic peptide of the same sequence was shown to be biologically active in small quantities and to extend the range of conditions suitable for development of competence. Cognate codons in the pneumococcal chromosome indicate that the peptide is made ribosomally. As the gene encodes a prepeptide containing the Gly-Gly consensus processing site found in peptide bacteriocins, the peptide is likely to be exported by a specialized ATP-binding cassette transport protein as is characteristic of these bacteriocins. The hypothesis is presented that this transport protein is encoded by comA, previously shown to be required for elaboration of the pneumococcal competence activator.


Safety and immunogenicity of the ChAdOx1 nCoV-19 (AZD1222) vaccine in children aged 6-17 years: a preliminary report of COV006, a phase 2 single-blind, randomised, controlled trial.
Vaccination of children and young people against SARS-CoV-2 is recommended in some countries. Scarce data have been published on immune responses induced by COVID-19 vaccines in people younger than 18 years compared with the same data that are available in adults. COV006 is a phase 2, single-blind, randomised, controlled trial of ChAdOx1 nCoV-19 (AZD1222) in children and adolescents at four trial sites in the UK. Healthy participants aged 6-17 years, who did not have a history of chronic respiratory conditions, laboratory-confirmed COVID-19, or previously received capsular group B meningococcal vaccine (the control), were randomly assigned to four groups (4:1:4:1) to receive two intramuscular doses of 5 × 1010 viral particles of ChAdOx1 nCoV-19 or control, 28 days or 84 days apart. Participants, clinical investigators, and the laboratory team were masked to treatment allocation. Study groups were stratified by age, and participants aged 12-17 years were enrolled before those aged 6-11 years. Due to the restrictions in the use of ChAdOx1 nCoV-19 in people younger than 30 years that were introduced during the study, only participants aged 12-17 years who were randomly assigned to the 28-day interval group had received their vaccinations at the intended interval (day 28). The remaining participants received their second dose at day 112. The primary outcome was assessment of safety and tolerability in the safety population, which included all participants who received at least one dose of the study drug. The secondary outcome was immunogenicity, which was assessed in participants who were seronegative to the nucleocapsid protein at baseline and received both prime and boost vaccine. This study is registered with ISRCTN (15638344). Between Feb 15 and April 2, 2021, 262 participants (150 [57%] participants aged 12-17 years and 112 [43%] aged 6-11 years; due to the change in the UK vaccination policy, the study terminated recruitment of the younger age group before the planned number of participants had been enrolled) were randomly assigned to receive vaccination with two doses of either ChAdOx1 nCoV-19 (n=211 [n=105 at day 28 and n=106 at day 84]) or control (n=51 [n=26 at day 28 and n=25 at day 84]). One participant in the ChAdOx1 nCoV-19 day 28 group in the younger age bracket withdrew their consent before receiving a first dose. Of the participants who received ChAdOx1 nCoV-19, 169 (80%) of 210 participants reported at least one solicited local or systemic adverse event up to 7 days following the first dose, and 146 (76%) of 193 participants following the second dose. No serious adverse events related to ChAdOx1 nCoV-19 administration were recorded by the data cutoff date on Oct 28, 2021. Of the participants who received at least one dose of ChAdOx1 nCoV-19, there were 128 unsolicited adverse events up to 28 days after vaccination reported by 83 (40%) of 210 participants. One participant aged 6-11 years receiving ChAdOx1 nCoV-19 reported a grade 4 fever of 40·2°C on day 1 following first vaccination, which resolved within 24 h. Pain and tenderness were the most common local solicited adverse events for all the ChAdOx1 nCoV-19 and capsular group B meningococcal groups following both doses. Of the 242 participants with available serostatus data, 14 (6%) were seropositive at baseline. Serostatus data were not available for 20 (8%) of 262 participants. Among seronegative participants who received ChAdOx1 nCoV-19, anti-SARS-CoV-2 IgG and pseudoneutralising antibody titres at day 28 after the second dose were higher in participants aged 12-17 years with a longer interval between doses (geometric means of 73 371 arbitrary units [AU]/mL [95% CI 58 685-91 733] and 299 half-maximal inhibitory concentration [IC50; 95% CI 230-390]) compared with those aged 12-17 years who received their vaccines 28 days apart (43 280 AU/mL [95% CI 35 852-52 246] and 150 IC50 [95% CI 116-194]). Humoral responses were higher in those aged 6-11 years than in those aged 12-17 years receiving their second dose at the same 112-day interval (geometric mean ratios 1·48 [95% CI 1·07-2·07] for anti-SARS-CoV-2 IgG and 2·96 [1·89-4·62] for pseudoneutralising antibody titres). Cellular responses peaked after a first dose of ChAdOx1 nCoV-19 across all age and interval groups and remained above baseline after a second vaccination. ChAdOx1 nCoV-19 is well tolerated and immunogenic in children aged 6-17 years, inducing concentrations of antibody that are similar to those associated with high efficacy in phase 3 studies in adults. No safety concerns were raised in this trial. AstraZeneca and the UK Department of Health and Social Care through the UK National Institute for Health and Care Research.


Tracking sensory system atrophy and outcome prediction in spinal cord injury.
In patients with subacute spinal cord injury (SCI), the motor system undergoes progressive structural changes rostral to the lesion, which are associated with motor outcome. The extent to which the sensory system is affected and how this relates to sensory outcome are uncertain. Changes in the sensory system were prospectively followed by applying a comprehensive magnetic resonance imaging (MRI) protocol to 14 patients with subacute traumatic SCI at baseline, 2 months, 6 months, and 12 months after injury, combined with a full neurological examination and comprehensive pain assessment. Eighteen controls underwent the same MRI protocol. T1-weighted volumes, myelin-sensitive magnetization transfer saturation (MT), and longitudinal relaxation rate (R1) mapping provided data on spinal cord and brain morphometry and microstructure. Regression analysis assessed the relationship between MRI readouts and sensory outcomes. At 12 months from baseline, sensory scores were unchanged and below-level neuropathic pain became prominent. Compared with controls, patients showed progressive degenerative changes in cervical cord and brain morphometry across the sensory system. At 12 months, MT and R1 were reduced in areas of structural decline. Sensory scores at 12 months correlated with rate of change in cord area and brain volume and decreased MT in the spinal cord at 12 months. This study has demonstrated progressive atrophic and microstructural changes across the sensory system with a close relation to sensory outcome. Structural MRI protocols remote from the site of lesion provide new insights into neuronal degeneration underpinning sensory disturbance and have potential as responsive biomarkers of rehabilitation and treatment interventions.