Periprocedural complication rates are equivalent between symptomatic and asymptomatic patients undergoing carotid angioplasty and stenting.
Patients with neurologic symptoms who undergo carotid endarterectomy (CEA) have a higher incidence of stroke and death in the perioperative period than those with asymptomatic carotid disease. This study examines the outcomes of symptomatic and asymptomatic patients undergoing carotid stenting (CAS). From 2002 to 2006, 201 CAS procedures were performed in 193 patients (117 men, mean age 73 +/- 10 years), of whom 142 were for asymptomatic (AS) and 59 for symptomatic (S) disease. Preoperative neurologic symptoms included recent ipsilateral cerebrovascular accident (CVA, 29%), transient ischemic attack (50%), and amaurosis fugax (22%). There were 201 carotid stents placed (107 Acculink, 43 Wallstent, 23 Precise, 21 NexStent, 3 Exponent, 3 Xact, 1 Herculink) and 198 protection devices used (79 Accunet, 53 EPI Filterwire, 43 PercuSurge, 20 Angiogard, 3 EmboShield). Mean follow-up was 41 weeks. The groups were matched in terms of demographics and comorbidities (carotid artery disease, hypertension, hyperlipidemia, diabetes mellitus, peripheral vascular disease, smoking, and chronic obstructive pulmonary disease; p = nonsignificant [NS]). There was no significant difference in anatomic risk factors (neck irradiation, S 3%, AS 6%; prior CEA, S 14%, AS 14%; bovine arch, S 22%, AS 16%; p = NS), and the types of embolic protection devices and stents used were similar between groups. The mean percentages of preintervention carotid stenosis were equal (S 88%, AS 88%), and the technical success rate was 99%. Incidence rates of CVA (S 3.4%, AS 1.4%), myocardial infarction (S 1.7%, AS 1.4%), and death (S 0, AS 0.7%) were equivalent between groups (p = NS). CAS with cerebral protection can be performed safely in both symptomatic and asymptomatic patients. The presence of preoperative neurologic symptoms does not significantly increase the risk of adverse events in the perioperative period in this study.


Expression of foetal type acetylcholine receptor is restricted to type 1 muscle fibres in human neuromuscular disorders.
In adult muscle, acetylcholine receptors (AChR) are restricted mainly to the motor endplate where the adult isoform (alphabetadeltaepsilon) is expressed. When skeletal muscle is denervated in animal models, there is atrophy of the muscle and a marked increase in expression of the AChR foetal isoform (alphabetagammadelta) containing a gamma-subunit. Similar changes in AChR expression are thought to occur in human muscle. While the role of denervation in regulating AChR gene expression has been widely studied, it has not been determined whether the transcriptional programmes responsible for defining different fibre types have an impact on the expression of AChR genes. We investigated biopsies from patients with a wide spectrum of neuromuscular diseases for expression of the AChR alpha- and gamma-subunits using RNase protection assays, alpha/gamma-duplex reverse transcriptase polymerase chain reaction, immunohistochemistry for foetal AChR and RNA in situ hybridization. Muscle from all patients with neurogenic disorders and, to a lesser extent, myogenic disorders, exhibited markedly increased transcription of the AChR gamma-subunit but, in contrast to previous animal studies, did not show increased AChR alpha-subunit. Moreover, both immunohistochemistry and RNA in situ hybridization revealed that AChR gamma-subunit hyperexpression occurred exclusively in atrophic type 1 and not in atrophic type 2 muscle fibres, irrespective of the underlying neuromuscular disease. We conclude that up-regulation of the AChR gamma-subunit in human muscle disorders is restricted to type 1 muscle fibres and, therefore, that AChR gamma-subunit expression is controlled by a muscle fibre type-restricted transcriptional programme. The factors influencing expression of this and other functional proteins should be relevant to the understanding and treatment of a range of neuromuscular disorders.


A molecule targeting VHL-deficient renal cell carcinoma that induces autophagy.
Renal cell carcinomas (RCCs) are refractory to standard therapies. The von Hippel-Lindau (VHL) tumor suppressor gene is inactivated in 75% of RCCs. By screening for small molecules selectively targeting VHL-deficient RCC cells, we identified STF-62247. STF-62247 induces cytotoxicity and reduces tumor growth of VHL-deficient RCC cells compared to genetically matched cells with wild-type VHL. STF-62247-stimulated toxicity occurs in a HIF-independent manner through autophagy. Reduction of protein levels of essential autophagy pathway components reduces sensitivity of VHL-deficient cells to STF-62247. Using a yeast deletion pool, we show that loss of proteins involved in Golgi trafficking increases killing by STF-62247. Thus, we have found a small molecule that selectively induces cell death in VHL-deficient cells, representing a paradigm shift for targeted therapy.


Comparison of prior authorization and prospective audit with feedback for antimicrobial stewardship.
Although prior authorization and prospective audit with feedback are both effective antimicrobial stewardship program (ASP) strategies, the relative impact of these approaches remains unclear. We compared these core ASP strategies at an academic medical center. Quasi-experimental study. We compared antimicrobial use during the 24 months before and after implementation of an ASP strategy change. The ASP used prior authorization alone during the preintervention period, June 2007 through May 2009. In June 2009, many antimicrobials were unrestricted and prospective audit was implemented for cefepime, piperacillin/tazobactam, and vancomycin, marking the start of the postintervention period, July 2009 through June 2011. All adult inpatients who received more than or equal to 1 dose of an antimicrobial were included. The primary end point was antimicrobial consumption in days of therapy per 1,000 patient-days (DOT/1,000-PD). Secondary end points included length of stay (LOS). In total, 55,336 patients were included (29,660 preintervention and 25,676 postintervention). During the preintervention period, both total systemic antimicrobial use (-9.75 DOT/1,000-PD per month) and broad-spectrum anti-gram-negative antimicrobial use (-4.00 DOT/1,000-PD) declined. After the introduction of prospective audit with feedback, however, both total antimicrobial use (+9.65 DOT/1,000-PD per month; P < .001) and broad-spectrum anti-gram-negative antimicrobial use (+4.80 DOT/1,000-PD per month; P < .001) increased significantly. Use of cefepime and piperacillin/tazobactam both significantly increased after the intervention (P = .03). Hospital LOS and LOS after first antimicrobial dose also significantly increased after the intervention (P = .016 and .004, respectively). Significant increases in antimicrobial consumption and LOS were observed after the change in ASP strategy.


Swelling, reductive stress, and cell death during chemical hypoxia in hepatocytes.
In rat hepatocytes, we examined the relationship between cell volume, bleb formation, and loss of cell viability during chemical hypoxia with KCN plus iodoacetic acid. In hypotonic media (150-200 mosmol/kgH2O), cells swelled to a greater extent during chemical hypoxia than in isotonic media, but rates of cell killing were identical. Sucrose (300 mM) added to isotonic media prevented early cell swelling but actually accelerated cell killing. In contrast, mannitol (300 mM) improved cell survival but did not prevent cell swelling. Bleb formation occurred regardless of buffer tonicity. The antioxidants desferrioxamine and cyanidanol but not superoxide dismutase +/- catalase delayed lethal cell injury. Cell killing was greater during aerobic compared with anaerobic chemical hypoxia. Hydroperoxide formation was measured using a dichlorofluorescin assay and was accelerated during aerobic but not anaerobic chemical hypoxia. The results indicate that cell swelling is not the driving force for bleb formation or lethal cell injury. We conclude that "reductive stress" caused by respiratory inhibition favors formation of toxic oxygen species and may contribute to lethal cell injury during intermittent or incomplete oxygen deprivation.


Membrane release and destabilization of Arabidopsis RIN4 following cleavage by Pseudomonas syringae AvrRpt2.
The Arabidopsis RIN4 protein mediates interaction between the Pseudomonas syringae type III effector proteins AvrB, AvrRpm1, and AvrRpt2 and the Arabidopsis disease-resistance proteins RPM1 and RPS2. Confocal laser-scanning fluorescence microscopy following particle bombardment of tobacco leaf epidermal cells was used to examine the subcellular localization of fusions between GFP and RIN4 or several of its homologs and to examine the effects of cobombardment with AvrRpt2 or AvrRpml. This study showed that RIN4 was attached to the plasma membrane at its carboxyl terminus and that a carboxyl-terminal CCCFxFxxx prenylation or acylation (typically palmitoylation) motif, or both, was essential for this attachment. RIN4 was cleaved by AvrRpt2 at two PxFGxW motifs, one releasing a large portion of RIN4 from the plasma membrane and both exposing amino-terminal residues that destabilized the carboxyl-terminal cleavage products by targeting them for N-end ubiquitylation and proteasomal degradation. Plasma-membrane localization of RIN4 was not affected by AvrRpml. RIN4 was found to be part of a protein family comprising two full-length homologs and at least 11 short carboxyl-terminal homologs. Representatives of this family, comprising a full-length RIN4 homolog and two short carboxyl-terminal RIN4 homologs, were also attached to the plasma membrane and cleaved near their amino termini by AvrRpt2, but in contrast to RIN4, the major portions of these proteins remained on the plasma membrane. N-end degradation may play a minor role in RIN4 degradation but probably plays a major role in the degradation of RIN4 homologs and is, therefore, a major pathogenic consequence of AvrRpt2 cleavage.


Making the case for a fracture liaison service: a qualitative study of the experiences of clinicians and service managers.
To develop services, healthcare professionals must make business cases to managerial bodies within Hospital Trusts and if approved, to commissioning bodies. Patients with hip fracture are at high risk of subsequent fracture. To prevent this, guidance recommends structuring fracture prevention services around coordinator based models. These are known as Fracture Liaison Services (FLS). 33 semi-structured qualitative interviews were conducted with healthcare professionals with experience of making business cases for FLS. Data was analysed thematically. Challenges in the development of business cases included collecting all the relevant data and negotiating compartmentalised budgets that impeded service development. Participants described communication and cooperation between providers and commissioners as variable. They felt financial considerations were the most important factor in funding decisions, while improved quality of care was less influential. Other factors included national guidelines and political priorities. The personalities of clinicians championing services, and the clinical interests of commissioners were seen to influence the decision-making process, suggesting that participants felt that decisions were not always made on the basis of evidence-based care. Effective strategies included ways of providing support, demonstrating potential cost effectiveness and improved quality of care. Using a range of sources including audit data collected on the successful Glasgow FLS, and improving cooperation between stakeholders was advocated. Participants felt that the work of commissioners and providers should be better integrated and suggested strategies for doing this. This study provides information to healthcare professionals about how best to develop business cases for FLS. We conclude with recommendations on how to develop effective cases. These include using guidance such as toolkits, aligning the aims of FLS with national priorities and benchmarking services against comparators. Introducing a 'Local Champion' to work alongside the service manager and establishing a multi-disciplinary working team would facilitate communication between stakeholders. Involving commissioners in service design would help integrate the roles of purchasers and providers.


Ru2 and Ru encode mouse orthologs of the genes mutated in human Hermansky-Pudlak syndrome types 5 and 6.
Hermansky-Pudlak syndrome (HPS) is a genetically heterogeneous disease involving abnormalities of melanosomes, platelet dense granules and lysosomes. Here we have used positional candidate and transgenic rescue approaches to identify the genes mutated in ruby-eye 2 and ruby-eye mice (ru2 and ru, respectively), two 'mimic' mouse models of HPS. We also show that these genes are orthologs of the genes mutated in individuals with HPS types 5 and 6, respectively, and that their protein products directly interact. Both genes are previously unknown and are found only in higher eukaryotes, and together represent a new class of genes that have evolved in higher organisms to govern the synthesis of highly specialized lysosome-related organelles.


When are medical apps medical? Off-label use and the Food and Drug Administration.
People have a love/hate relationship with rapidly changing healthcare technology. While consumer demand for medical apps continues to grow as rapidly as does supply (there are over 100,000 health, wellness and medical applications, or 'apps' on the market), healthcare professionals and safety experts worry about the impact of these apps on the health consumer. In response to the rapidly growing mobile healthcare sector, the Food and Drug Administration has put forth guidelines to regulate 'mobile medical apps' (MMAs), those health-related apps that are (self) designated as medical devices. In this article, I argue that this decision, to only regulate apps that bill themselves as medical devices, will create a market for 'off-label' app use. Further, I will talk about the oft used analogy between off-label apps and off-label pharmaceuticals, showing that off-labeling apps will provide patients none of the benefits that come with a physician prescribing a drug off-label, while exposing the mobile healthcare consumer to significant risks that go significantly beyond those that we know of (and must accept) from prescription drugs. Recognizing that the Food and Drug Administration is not going to be able to significantly change its policies on oversight, I will suggest specific actions to at least mitigate some of the risks associated with off-label app use.


AlF3-Modified carbon nanofibers as a multifunctional 3D interlayer for stable lithium metal anodes.
A versatile interlayer in which AlF3 particles are embedded within carbon nanofibers (AlF3@CNFs) is reported to induce controllable Li+ deposition and achieve a dendrite free Li metal anode. Due to the AlF3 reducing Li nucleation overpotential and CNFs providing high Young's moduli up to 47 GPa, a high Coulombic efficiency (97.2%) and a long lifespan (900 h) are achieved in the carbonate-based electrolyte. A long cycling life is also realized by using interlayer-protected lithium anodes in Li-S batteries.


Zolfino landrace (Phaseolus vulgaris L.) from Pratomagno: general and specific features of a functional food.
The Zolfino bean is a variety of Phaseolus vulgaris, which is cultivated in a limited area of Tuscany, Italy, and is widely appreciated for its flavor and culinary uses. A yellow Zolfino landrace cultivated in the Leccio-Reggello area was characterized and compared with three other varieties of Phaseolus vulgaris (i.e. the Borlotto, Cannellino, and Corona beans) in terms of its general features and potential as an antioxidant/anti-inflammatory agent. The length, width, thickness, equatorial section surface, weight, volume, and seed coat section were measured in all the beans. The seed surface area was also estimated by an original empirical method. The ability of the different beans to interfere with the enzymes of the polyol pathway (that is, aldose reductase (AR) and sorbitol dehydrogenase) was tested using the supernatant after soaking the beans at room temperature and after thermal treatment, which simulated the bean-cooking process in a controlled fashion. Concerning the general features, Zolfino was comparable with other beans, except Corona, in terms of surface-volume ratio, which possesses the lowest tegument thickness. Moreover, Zolfino appears the most effective in inhibiting AR activity. The inhibitory ability is unaffected by thermal treatment and appears to be associated with compound(s) present in the coat of the bean. The ability of Zolfino to inhibit AR, thus reducing the flux of glucose through the polyol pathway, highlights the features of Zolfino as a functional food, potentially useful in treating the dysfunctions linked to the hyperactivity of AR, such as diabetic complications or inflammatory responses.


Build Your Own Pension: Framing Pension Reform and Choice in Newspapers.
Recent pension reforms in Norway mean that old-age pensions to a greater extent are a function of adaptations made and decisions made throughout the lifetime. How much you work, who you work for, when you retire, and how you invest will influence your standard of living as an old-age pensioner. This article investigates one important source of information about retirement and pension, namely service journalism in main national newspapers. How is the issue of pension framed in these articles? And what kind of advice is presented by which kind of actors? Findings indicate that service journalism is dominated by pension industry sources, and old-age pension is framed as a function of individual investment choices rather than as citizens' rights.


Evaluation of the impact of two educational interventions on GP management of familial breast/ovarian cancer cases: a cluster randomised controlled trial.
It has been suggested that primary care should become more involved in providing genetic services, such as basic risk assessment, to enable patients with a moderate/high risk to be referred and those with a low risk to receive reassurance and advice from their general practitioner (GP). However, GPs currently lack knowledge and confidence in this area. To investigate the effect of an in-practice educational session and information pack on GP management of familial breast/ovarian cancer cases. Cluster randomised controlled trial. GP principals in 170 practices in Oxfordshire and Northamptonshire. Of the 688 GPs, 426 (62%) participated. Practices were randomised either to Group A (receiving an inpractice educational session plus information pack), Group B (receiving an information pack alone), or Group C (receiving neither an educational session nor a pack). The main study outcome was the proportion of GPs making the correct referral decision on at least five out of six family history vignettes. A secondary outcome was GPs' reported confidence in managing patients with a family history of breast/ovarian cancer, measured by a score that was generated by combining responses to four questions. There was a 40% (95% CI = 30-50%, P < 0.001) improvement in the proportion of GPs who made the correct referral decision on at least five out of the six vignettes in Group A (111/140 [79%]) compared with Group C (controls) (63/162 [39%]), and a 42% (95% CI = 31-52%, P < 0.001) improvement in Group B (100/124 [81%]) compared with the control group. There was a trend in reported confidence in the management of individuals with a family history of breast/ovarian cancer from a mean confidence score of 2.3 in Group A to 2.0 in Group B and 1.5 in Group C (P < 0.001). Providing GPs with an information pack significantly improved referral decisions regarding patients with a family history of breast/ovarian cancer. Although extremely well received, an in-house educational session produced no additional improvements. There were, however, greater levels of reported confidence in the group who received the educational session in addition to the information pack.


Modeling the role of public health education in Ebola virus disease outbreaks in Sudan.
Public involvement in Ebola Virus Disease (EVD) prevention efforts is key to reducing disease outbreaks. Targeted education through practical health information to particular groups and sub-populations is crucial to controlling the disease. In this paper, we study the dynamics of Ebola virus disease in the presence of public health education with the aim of assessing the role of behavior change induced by health education to the dynamics of an outbreak. The power of behavior change is evident in two outbreaks of EVD that took place in Sudan only 3 years apart. The first occurrence was the first documented outbreak of EVD and produced a significant number of infections. The second outbreak produced far fewer cases, presumably because the population in the region learned from the first outbreak. We derive a system of ordinary differential equations to model these two contrasting behaviors. Since the population in Sudan learned from the first outbreak of EVD and changed their behavior prior to the second outbreak, we use data from these two instances of EVD to estimate parameters relevant to two contrasting behaviors. We then simulate a future outbreak of EVD in Sudan using our model that contains two susceptible populations, one being more informed about EVD. Our finding show how a more educated population results in fewer cases of EVD and highlights the importance of ongoing public health education.


The branchpoint binding protein: in and out of the spliceosome cycle.
The Saccharomyces cerevisiae branchpoint binding protein (BBP) is a 53 kDa pre-mRNA processing factor with characteristic STAR/GSG protein organization. This includes a central RNA binding site composed of an extended Type I KH domain with an adjacent QUA2 motif. Downstream of KH-QUA2 are two CCHC-type zinc knuckles and a proline-rich C-terminal interaction domain (Fig. 1A). The QUA1 homodimerization motif found upstream of the KH-QUA2 sequence in other STAR/GSG family members is absent in BBP and replaced by a site for the phylogenetically conserved binding partner, Mud2/U2AF65. BBP's name reflects the fact that it binds the conserved RNA sequence, UACUAAC, called the branchpoint motif found near the 3' end of yeast introns. This sequence contains the catalytic adenosine (underlined) which directs the first RNA transesterification reaction in splicing chemistry. BBP recruitment to the branchpoint initiates a series of spliceosomal subunit addition and rearrangement events that ultimately configures the active site ofthis enzyme. The mammalian homolog, ZFM1/ZNF162/D11S636/ SF1 (henceforth, SF1), was first identified in a screen for genes associated with Type 1 multiple endocrine neoplasia2 and was subsequently shown to act similarly to BBP in mammalian splicing. BBP/SF1 is essential for viability in organisms spanning the evolutionary spectrum from yeast to Caenorhabditis elegans to mice. In addition, mice heterozygous for a SF1 knockout allele show enhanced susceptibility to azoxymethane-induced colon tumorigenesis adding BBP/SF1 to the growing list of RNA processing factors implicated in genetic disease. Summarized below is our current understanding of BBP structure and its proposed multifaceted contribution to mRNA biogenesis and function. Reference to SF1 will be made to fill gaps in our understanding of BBP and to highlight areas of clear similarity or difference between yeast and mammals.


Engineering exon-skipping vectors expressing U7 snRNA constructs for Duchenne muscular dystrophy gene therapy.
Duchenne muscular dystrophy (DMD) is a fatal muscle wasting disorder caused by mutations in the dystrophin gene. In most cases, the open-reading frame is disrupted which results in the absence of a functional protein. Antisense-mediated exon skipping is one of the most promising approaches for the treatment of DMD and has recently been shown to correct the reading frame and restore dystrophin expression in vitro and in vivo. Specific exon skipping can be achieved using synthetic oligonucleotides or viral -vectors encoding modified snRNAs, by masking important splicing sites. We have recently demonstrated that enhanced exon skipping can be induced by a U7 snRNA carrying binding sites for the heterogeneous ribonucleoprotein A1. In DMD patient cells, bifunctional U7 snRNAs harboring silencer motifs induce complete skipping of exon 51 and thus restore dystrophin expression to near wild-type levels. Furthermore, we have confirmed the efficacy of these constructs in vivo in transgenic mice carrying the entire human DMD locus after intramuscular injection of AAV vectors encoding the bifunctional U7 snRNA. These new constructs are very promising for the optimization of therapeutic exon skipping for DMD, but also offer powerful and versatile tools to modulate pre-mRNA splicing in a wide range of applications. Here, we outline the design of these U7 snRNA constructs to achieve efficient exon skipping of the dystrophin gene. We also describe methods to evaluate the efficiency of such U7 snRNA constructs in vitro in DMD patient cells and in vivo in the transgenic hDMD mouse model, using lentiviral and recombinant adeno-associated viral vectors, respectively.


Alteration of gray matter microstructure in schizophrenia.
Neuroimaging studies demonstrate gray matter (GM) macrostructural abnormalities in patients with schizophrenia (SCZ). While ex-vivo and genetic studies suggest cellular pathology associated with abnormal neurodevelopmental processes in SCZ, few in-vivo measures have been proposed to target microstructural GM organization. Here, we use diffusion heterogeneity- to study GM microstructure in SCZ. Structural and diffusion magnetic resonance imaging (MRI) were acquired on a 3 Tesla scanner in 46 patients with SCZ and 37 matched healthy controls (HC). After correction for free water, diffusion heterogeneity as well as commonly used diffusion measures FA and MD and volume were calculated for the four cortical lobes on each hemisphere, and compared between groups. Patients with early course SCZ exhibited higher diffusion heterogeneity in the GM of the frontal lobes compared to controls. Diffusion heterogeneity of the frontal lobe showed excellent discrimination between patients and HC, while none of the commonly used diffusion measures such as FA or MD did. Higher diffusion heterogeneity in the frontal lobes in early SCZ may be due to abnormal brain maturation (migration, pruning) before and during adolescence and early adulthood. Further studies are needed to investigate the role of heterogeneity as potential biomarker for SCZ risk.


Endocardial to myocardial notch-wnt-bmp axis regulates early heart valve development.
Endocardial to mesenchymal transformation (EMT) is a fundamental cellular process required for heart valve formation. Notch, Wnt and Bmp pathways are known to regulate this process. To further address how these pathways coordinate in the process, we specifically disrupted Notch1 or Jagged1 in the endocardium of mouse embryonic hearts and showed that Jagged1-Notch1 signaling in the endocardium is essential for EMT and early valvular cushion formation. qPCR and RNA in situ hybridization assays reveal that endocardial Jagged1-Notch1 signaling regulates Wnt4 expression in the atrioventricular canal (AVC) endocardium and Bmp2 in the AVC myocardium. Whole embryo cultures treated with Wnt4 or Wnt inhibitory factor 1 (Wif1) show that Bmp2 expression in the AVC myocardium is dependent on Wnt activity; Wnt4 also reinstates Bmp2 expression in the AVC myocardium of endocardial Notch1 null embryos. Furthermore, while both Wnt4 and Bmp2 rescue the defective EMT resulting from Notch inhibition, Wnt4 requires Bmp for its action. These results demonstrate that Jagged1-Notch1 signaling in endocardial cells induces the expression of Wnt4, which subsequently acts as a paracrine factor to upregulate Bmp2 expression in the adjacent AVC myocardium to signal EMT.


Evolution of developmental mechanisms in plants.
As our understanding of developmental mechanisms in flowering plant species has become more advanced, an appreciation of the need to understand how distinct plant morphologies are generated has grown. This has led to an awareness of the key morphological differences in distinct land plant groups and to an assessment of the major innovations that occurred during land plant evolution. Recent advances demonstrate how developmental toolkits have been recruited for related purposes in different land plant groups, but the limited number of examples highlights both the infancy of the field and the difficulty of working with non-flowering plants.


Morphometric analysis of human peripapillary retinal nerve fiber layer thickness.
The peripapillary retinal nerve fiber layer (RNFL) thickness pattern in the normal human eye has been well characterized using data obtained with scanning laser polarimetry and optical coherence tomography. The authors sought to characterize the normative peripapillary RNFL thickness and pattern using histologic sections obtained from healthy postmortem human eyes. Seventeen unpaired normal postmortem eyes were recruited into this study. Each eye was sectioned using the "umbrella technique" to obtain four concentric peripapillary rings, each centered on the optic disc, with diameters of 3.0, 3.5, 4.0, and 4.5 mm. RNFL thickness data along each ring section was measured at 100 equidistant locations. Thickness data, for each ring diameter, across all eyes were averaged to arrive at normative thickness values for the peripapillary RNFL thickness in eyes processed using this technique. Average RNFL thicknesses (+/-SD) for the 3.5-mm diameter ring were as follows: overall, 60.3 +/- 19.5 microm; superior, 75.3 +/- 26.5 mum; inferior, 69.4 +/- 22.4 microm; nasal, 48.1 +/- 15 microm; temporal, 49.2 +/- 26.4 microm. Qualitatively, the RNFL thickness showed a double-hump pattern with relatively similar superior and inferior peaks and with temporal and nasal troughs. Progressively larger peripapillary rings showed progressively thinner RNFL thickness at all quadrants. In contrast, the relative thickness percentage for each quadrant remained unchanged among the four different diameter rings. Histologic data from a group of healthy postmortem eyes demonstrate the pattern of RNFL thickness in normal eyes. These data corroborate imaging findings of peripapillary RNFL thickness patterns obtained using commercially available RNFL imaging devices.


Immunogenetics and genomics.
Immunogenetic analysis of disease susceptibility has been encouraged by the identification of strong HLA associations with several diseases of uncertain cause. Weaker HLA associations exist with a large number of infectious and non-infectious diseases and the mechanisms of these effects are beginning to be uncovered. Extensive analyses of non-HLA immunogenetic variants have also been undertaken and associations with a variety of genes identified. Genetic linkage analysis of multicase families has recently identified new major susceptibility loci for a few immunologically determined common diseases. However, the greatest potential for the future lies in genome-wide searches for susceptibility genes that individually might have quite modest effects but cumulatively have a large impact on individual risk. This new era of immunogenomics promises to provide key insights into disease pathogenesis and identify multiple molecular targets for intervention strategies.


The chicken chorioallantoic membrane model in biology, medicine and bioengineering.
The chicken chorioallantoic membrane (CAM) is a simple, highly vascularized extraembryonic membrane, which performs multiple functions during embryonic development, including but not restricted to gas exchange. Over the last two decades, interest in the CAM as a robust experimental platform to study blood vessels has been shared by specialists working in bioengineering, development, morphology, biochemistry, transplant biology, cancer research and drug development. The tissue composition and accessibility of the CAM for experimental manipulation, makes it an attractive preclinical in vivo model for drug screening and/or for studies of vascular growth. In this article we provide a detailed review of the use of the CAM to study vascular biology and response of blood vessels to a variety of agonists. We also present distinct cultivation protocols discussing their advantages and limitations and provide a summarized update on the use of the CAM in vascular imaging, drug delivery, pharmacokinetics and toxicology.


Deconvolution of High Dimensional Mixtures via Boosting, with Application to Diffusion-Weighted MRI of Human Brain.
Diffusion-weighted magnetic resonance imaging (DWI) and fiber tractography are the only methods to measure the structure of the white matter in the living human brain. The diffusion signal has been modelled as the combined contribution from many individual fascicles of nerve fibers passing through each location in the white matter. Typically, this is done via basis pursuit, but estimation of the exact directions is limited due to discretization [1, 2]. The difficulties inherent in modeling DWI data are shared by many other problems involving fitting non-parametric mixture models. Ekanadaham et al. [3] proposed an approach, continuous basis pursuit, to overcome discretization error in the 1-dimensional case (e.g., spike-sorting). Here, we propose a more general algorithm that fits mixture models of any dimensionality without discretization. Our algorithm uses the principles of L2-boost [4], together with refitting of the weights and pruning of the parameters. The addition of these steps to L2-boost both accelerates the algorithm and assures its accuracy. We refer to the resulting algorithm as elastic basis pursuit, or EBP, since it expands and contracts the active set of kernels as needed. We show that in contrast to existing approaches to fitting mixtures, our boosting framework (1) enables the selection of the optimal bias-variance tradeoff along the solution path, and (2) scales with high-dimensional problems. In simulations of DWI, we find that EBP yields better parameter estimates than a non-negative least squares (NNLS) approach, or the standard model used in DWI, the tensor model, which serves as the basis for diffusion tensor imaging (DTI) [5]. We demonstrate the utility of the method in DWI data acquired in parts of the brain containing crossings of multiple fascicles of nerve fibers.


Onset of small intestinal atrophy is associated with reduced intestinal blood flow in TPN-fed neonatal piglets.
Our aim was to determine the speed of onset of total parenteral nutrition (TPN)-induced mucosal atrophy, and whether this is associated with changes in intestinal blood flow and tissue metabolism in neonatal piglets. Piglets were implanted with jugular venous and duodenal catheters and either a portal venous or superior mesenteric artery (SMA) blood flow probe. At 3 wk of age, piglets were randomly assigned to receive continuous enteral formula feeding (n = 8) or TPN (n = 17) for 24 or 48 h. Blood flow was recorded continuously and piglets were given an i.v. bolus of bromodeoxyuridine and (13)C-phenylalanine to measure crypt cell proliferation and protein synthesis, respectively. After 8 h of TPN, portal and SMA blood flow decreased 30% compared with enteral feeding (P < 0.01), and remained near levels of food-deprived piglets for the remaining 48 h of TPN. After 24 h, TPN reduced jejunal inducible nitric oxide synthase (iNOS) activity and protein abundance (P < 0.05), small intestinal weight, and villous height (P < 0.01) compared with enterally fed piglets. Cell proliferation and DNA mass were decreased (P < 0.05) and apoptosis increased (P < 0.05) after 48 h of TPN. Protein synthesis was lower (P < 0.05) after 24 h of TPN, and protein mass was lower (P < 0.05) after 48 h of TPN, compared with enteral feeding. These data indicate that the transition from enteral to parenteral nutrition induced a rapid (<8 h) decrease in intestinal blood flow, and this likely precedes villous atrophy and the suppression of protein synthesis at 24 h, and of cell proliferation and survival at 48 h.


Weeding Out the Problem: The Impact of Preoperative Cannabinoid Use on Pain in the Perioperative Period.
The recreational and medical use of cannabinoids has been increasing. While most studies and reviews have focused on the role of cannabinoids in the management of acute pain, no study has examined the postoperative outcomes of surgical candidates who are on cannabinoids preoperatively. This retrospective cohort study examined the impact of preoperative cannabinoid use on postoperative pain scores and pain-related outcomes in patients undergoing major orthopedic surgery. Outcomes of patients who had major orthopedic surgery at our hospital between April 1, 2015 and June 30, 2017 were reviewed. Data were obtained from Networked Online Processing of Acute Pain Information, a locally developed database for our Acute Pain Service. Propensity score matching was used to balance baselines variables including age, sex, type of surgery, history of depression or anxiety, and perioperative use of regional anesthesia between patients who reported use of cannabinoids and those not on this substance. Intensity of pain with movement in the early postoperative period (defined as up to 36 hours after surgery) was the primary outcome of this study. The secondary outcomes (all in early postoperative period) were pain at rest, opioid consumption, incidence of pruritus, nausea and vomiting, sedation, delirium, constipation, impairment of sleep and physical activity, patient satisfaction with analgesia, and the length of Acute Pain Service follow-up. A total of 3793 patients were included in the study. Of these, 155 patients were identified as being on cannabinoids for recreational or medical indications in the preoperative period. After propensity score matching, we compared data from 155 patients who were on cannabinoids and 155 patients who were not on cannabinoids. Patients who were on preoperative cannabinoids had higher pain numerical rating score (median [25th, 75th percentiles]) at rest (5.0 [3.0, 6.1] vs 3.0 [2.0, 5.5], P = .010) and with movement (8.0 [6.0, 9.0] vs 7.0 [3.5, 8.5], P = .003), and a higher incidence of moderate-to-severe pain at rest (62.3% vs 45.5%, respectively, P = .004; odds ratio, 1.98; 95% CI, 1.25-3.14) and with movement (85.7% vs 75.2% respectively, P = .021; odds ratio, 1.98; 95% CI, 1.10-3.57) in the early postoperative period compared to patients who were not on cannabinoids. There was also a higher incidence of sleep interruption in the early postoperative period for patients who used cannabinoids. This retrospective study with propensity-matched cohorts showed that cannabinoid use was associated with higher pain scores and a poorer quality of sleep in the early postoperative period in patients undergoing major orthopedic surgery.


Model-based development of gemcabene, a new lipid-altering agent.
The purpose of this study was to evaluate the value of model-based, quantitative decision making during the development of gemcabene, a novel lipid-altering agent. The decisions were driven by a model of the likely clinical profile of gemcabene in comparison with its competitors, such as 3-hydroxymethylglutaryl coenzyme A reductase inhibitors (statins), the cholesterol absorption inhibitor ezetimibe, and their combination. Dose-response models were developed for the lipid effects (low-density lipoprotein cholesterol [LDL-C] and high-density lipoprotein cholesterol); adverse effects, such as persistent alanine aminotransferase elevation and myalgia; tolerability issues, such as headache; and risk reduction for coronary artery disease-related events for 5 statins, ezetimibe, gemcabene, and their combinations. The integrated model was based on the joint analysis of publicly available summary-level data and proprietary patient-level data and included information from almost 10,000 patients. The model was made available and accessible to the development team by using the Pharsight Drug Model Explorer model visualization technology. The modeling greatly enhanced the understanding of the clinical profile of gemcabene when given alone or in combination with a statin. The interaction between statins and gemcabene for the LDL-C lowering effect was found to be significantly different from the interaction between statins and ezetimibe. Ezetimibe was found to have a pharmacological-independent interaction resulting in additional LDL-C lowering over the entire statin dose range. The gemcabene interaction was found to be less than independent, resulting in almost no additional LDL-C lowering at high-statin doses, although the drug has a significant LDL-C effect when administered alone or in combination with a low dose of a statin. The quick availability of the model after completion of the first phase II trial in the target patient population and the ability of the team to explore the potential clinical efficacy and safety of gemcabene in comparison with alternative treatment options facilitated a quick decision to stop development.


Caveolin-1 tyrosine phosphorylation enhances paclitaxel-mediated cytotoxicity.
Caveolin-1 (CAV1), a highly conserved membrane-associated protein, is a putative regulator of cellular transformation. CAV1 is localized in the plasmalemma, secretory vesicles, Golgi, mitochondria, and endoplasmic reticulum membrane and associates with the microtubule cytoskeleton. Taxanes such as paclitaxel (Taxol) are potent anti-tumor agents that repress the dynamic instability of microtubules and arrest cells in the G(2)/M phase. Src phosphorylation of Tyr-14 on CAV1 regulates its cellular localization and function. We report that phosphorylation of CAV1 on Tyr-14 regulates paclitaxel-mediated apoptosis in MCF-7 breast cancer cells. Befitting its role as a multitasking molecule, we show that CAV1 sensitizes cells to apoptosis by regulating cell cycle progression and activation of the apoptotic signaling molecules BCL2, p53, and p21. We demonstrate that phosphorylated CAV1 triggers apoptosis by inactivating BCL2 and increasing mitochondrial permeability more efficiently than non-phosphorylated CAV1. Furthermore, expression of p21, which correlates with taxane sensitivity, is regulated by CAV1 phosphorylation in a p53-dependent manner. Collectively, our findings underscore the importance of CAV1 phosphorylation in apoptosis and suggest that events that negate CAV1 tyrosine phosphorylation may contribute to anti-microtubule drug resistance.


A Severe Outbreak of Charcoal Rot in Cantaloupe Melon Caused by Macrophomina phaseolina in Chile.
A severe outbreak of charcoal rot was observed in cantaloupe melon (Cucumis melo L.) in the summer of 2011 to 2012 in Curacaví Valley, Chile. Prior to harvest, of 72 plants per cultivar, charcoal rot prevalence varied from 32% to 82% in cvs. Colima, Charantias, Navigator, Origami, Otero, and Samoa. Symptoms were wilting and leaf browning associated with water-soaked lesions at the base of the crown with amber to dark brown exudates. Lesions dried out progressively, turned tan, and cracked. Affected plants declined and died before harvest. Reddish fruit decay was observed. Symptomatic stem and root samples (n = 97) were collected, surface disinfected (96% ethanol, 30 s), plated on PDA acidified with 0.5 ml/liter of 92% lactic acid (APDA), and incubated at 20 ± 1°C. A white, fast-growing mycelium was obtained that turned gray to black after 7 days due to the presence of spherical to oblong black microsclerotia 136 ± 52 μm (n = 80) in diameter. On the basis of colony morphology and microsclerotia, 57 isolates (59%), obtained from 97 melon samples, were tentatively identified as Macrophomina phaseolina (Tassi) Goid. (2,3). The morphological identification of four isolates M1HB-B, M2CO-B, M3CH-R, and M4OT-B (GenBank Accession Nos. JX203630, JX203631, JX203632, and JX203633) was confirmed by sequencing of the internal transcribed spacer region (ITS1-5.8S-ITS2) of rDNA, using primers ITS4 and ITS5, with >99% similarity with the sequences of M. phaseolina (GenBank Accession No. HQ660592) (4). Pathogenicity tests were conducted with isolates M1HB-B, M2CO-B, M3CH-R, and M4OT-B on melon fruits cvs. Colima, Origami, Charantias, and Diva. Four mature melon fruits per cultivar per isolate were surface disinfected with 0.5% sodium hypochlorite for 2 min before inserting a mycelium plug (19 mm2) in a 6 mm diameter hole made with a sterile cork borer. An equal number of perforated fruits in which a sterile agar plug was inserted were left as non-inoculated controls. After 8 days of incubation at 20°C, inoculated fruits developed a spherical, reddish, soft necrotic lesion of 15 to 20 mm in diameter in the pulp. Non-inoculated fruits remained symptomless. The pathogenicity of the four isolates was also studied in 3-month-old melon plants (n = 4) cvs. Colima and Navigator. Plants were inoculated by inserting a mycelial plug (9 mm2) underneath the epidermis of the crown, 5 cm above the soil level. The inoculation site was immediately wrapped with Parafilm to avoid dehydration. An equal number of non-inoculated, but injured plants, treated with a sterile agar plug, were left as controls. After 21 days of incubation under greenhouse conditions (17 ± 5.5°C), all inoculated plants developed water-soaked to dry necrotic lesions, 20 to 70 mm long, yellow to tan in color. No symptoms were obtained in non-inoculated controls. M. phaseolina was reisolated in 84% and 100% of the inoculated plants and fruits, respectively. To our knowledge, this study is the first report of charcoal rot in cantaloupe melon in Chile, previously found on watermelon and melon group inodorus (1). Charcoal rot appears as an emerging disease that aggressively affects current cantaloupe melon cultivars in central Chile. References: (1) G. Apablaza. Cien. Inv. Agr. 20:101, 1993. (2) B. D. Bruton and E. V. Wann. Charcoal rot. Page 9 in: Compendium of Cucurbit Diseases. T. A. Zitter, D. L. Hopkins, and C. E. Thomas, eds. APS, St. Paul, MN, 1996. (3) S. Kaur et al. Crit. Rev. Microbiol. 38:136, 2012. (4) J. Q. Zhang et al. Plant Dis. 95:872, 2011.


Accurate and Transferable Reactive Molecular Dynamics Models from Constrained Density Functional Theory.
Chemical reactions constitute the central feature of many liquid, material, and biomolecular processes. Conventional molecular dynamics (MD) is inadequate for simulating chemical reactions given the fixed bonding topology of most force fields, while modeling chemical reactions using ab initio molecular dynamics is limited to shorter time and length scales given its high computational cost. As such, the multiscale reactive molecular dynamics method provides one promising alternative for simulating complex chemical systems at atomistic detail on a reactive potential energy surface. However, the parametrization of such models is a key barrier to their applicability and success. In this work, we present reactive MD models derived from constrained density functional theory that are both accurate and transferable. We illustrate the features of these models for proton dissociation reactions of amino acids in both aqueous and protein environments. Specifically, we present models for ionizable glutamate and lysine that predict accurate absolute pKa values in water as well as their significantly shifted pKa in staphylococcal nuclease (SNase) without any modification of the models. As one outcome of the new methodology, the simulations show that the deprotonation of ionizable residues in SNase can be closely coupled with side chain rotations, which is a concept likely generalizable to many other proteins. Furthermore, the present approach is not limited to only pKa prediction but can enable the fully atomistic simulation of many other reactive systems along with a determination of the key aspects of the reaction mechanisms.


Unintentional firearm deaths in California.
We conducted a two-part study of unintentional firearm deaths in California. First, we analyzed death certificate data for the 688 unintentional firearm deaths of California residents occurring during 1977-1983. Mortality rates were 7.5 for males, 0.9 for females, 4.8 for whites, and 5.3 for blacks, all per million persons per year. Males ages 15-24 had the highest rate (17 per million persons per year). We then investigated the 131 childhood deaths in greater detail, using coroners' or medical examiners' reports. Most of these shootings occurred at a residence. Handguns were involved more frequently than predicted by their reported availability in homes in the region. Almost two thirds of child deaths resulted from head wounds, reinforcing the importance of primary prevention. At least 40% of child deaths in this study appeared in part to be attributable to defects in firearm performance or current firearm design practices, suggesting that improvements should be sought and mandated.


Thermal Destabilization of Collagen Matrix Hierarchical Structure by Freeze/Thaw.
This study aims to characterize and understand the effects of freezing on collagen structures and functionality. Specifically, thermodynamic destabilization of collagen at molecular- and fibril-levels by combination of low temperatures and freezing were experimentally characterized using modulated differential scanning calorimetry. In order to delineate the effects of sub-zero temperature and water-ice phase change, we hypothesized that the extent of destabilization can be determined based on post-thaw heat induced thermal denaturation of collagen. It is found that thermal denaturation temperature of collagen in hydrogel decreases by 1.4-1.6°C after freeze/thaw while no such decrease is observed in the case of molecular solution. The destabilization is predominantly due to ice formation. Exposure to low temperatures in the absence of ice has only minimal effect. Calorimetry measurements combined with morphological examination of collagen matrices by scanning electron microscopy suggest that freezing results in destabilization of collagen fibrils due to expansion of intrafibrillar space by ice formation. This fibril-level damage can be alleviated by use of cryoprotectant DMSO at concentrations as low as 0.5 M. A theoretical model explaining the change in collagen post-thaw thermal stability by freezing-induced fibril expansion is also proposed.


Climate change poised to threaten hydrologic connectivity and endemic fishes in dryland streams.
Protecting hydrologic connectivity of freshwater ecosystems is fundamental to ensuring species persistence, ecosystem integrity, and human well-being. More frequent and severe droughts associated with climate change are poised to significantly alter flow intermittence patterns and hydrologic connectivity in dryland streams of the American Southwest, with deleterious effects on highly endangered fishes. By integrating local-scale hydrologic modeling with emerging approaches in landscape ecology, we quantify fine-resolution, watershed-scale changes in habitat size, spacing, and connectance under forecasted climate change in the Verde River Basin, United States. Model simulations project annual zero-flow day frequency to increase by 27% by midcentury, with differential seasonal consequences on continuity (temporal continuity at discrete locations) and connectivity (spatial continuity within the network). A 17% increase in the frequency of stream drying events is expected throughout the network with associated increases in the duration of these events. Flowing portions of the river network will diminish between 8% and 20% in spring and early summer and become increasingly isolated by more frequent and longer stretches of dry channel fragments, thus limiting the opportunity for native fishes to access spawning habitats and seasonally available refuges. Model predictions suggest that midcentury and late century climate will reduce network-wide hydrologic connectivity for native fishes by 6-9% over the course of a year and up to 12-18% during spring spawning months. Our work quantifies climate-induced shifts in stream drying and connectivity across a large river network and demonstrates their implications for the persistence of a globally endemic fish fauna.


Late-surviving megafauna in Tasmania, Australia, implicate human involvement in their extinction.
Establishing the cause of past extinctions is critical if we are to understand better what might trigger future occurrences and how to prevent them. The mechanisms of continental late Pleistocene megafaunal extinction, however, are still fiercely contested. Potential factors contributing to their demise include climatic change, human impact, or some combination. On the Australian mainland, 90% of the megafauna became extinct by approximately 46 thousand years (ka) ago, soon after the first archaeological evidence for human colonization of the continent. Yet, on the neighboring island of Tasmania (which was connected to the mainland when sea levels were lower), megafaunal extinction appears to have taken place before the initial human arrival between 43 and 40 ka, which would seem to exonerate people as a contributing factor in the extirpation of the island megafauna. Age estimates for the last megafauna, however, are poorly constrained. Here, we show, by direct dating of fossil remains and their associated sediments, that some Tasmanian megafauna survived until at least 41 ka (i.e., after their extinction on the Australian mainland) and thus overlapped with humans. Furthermore, a vegetation record for Tasmania spanning the last 130 ka shows that no significant regional climatic or environmental change occurred between 43 and 37 ka, when a land bridge existed between Tasmania and the mainland. Our results are consistent with a model of human-induced extinction for the Tasmanian megafauna, most probably driven by hunting, and they reaffirm the value of islands adjacent to continental landmasses as tests of competing hypotheses for late Quaternary megafaunal extinctions.


Comprehensive Profiling of the Native and Modified Peptidomes of Raw Bovine Milk and Processed Milk Products.
Bovine milk contains a variety of endogenous peptides, partially formed by milk proteases that may exert diverse bioactive functions. Milk storage allows further protease activities altering the milk peptidome, while processing, e.g., heat treatment can trigger diverse chemical reactions, such as Maillard reactions and oxidations, leading to different posttranslational modifications (PTMs). The influence of processing on the native and modified peptidome was studied by analyzing peptides extracted from raw milk (RM), ultra-high temperature (UHT) milk, and powdered infant formula (IF) by nano reversed-phase liquid chromatography coupled online to electrospray ionization (ESI) tandem mass spectrometry. Only unmodified peptides proposed by two independent software tools were considered as identified. Thus, 801 identified peptides mainly originated from αS- and β-caseins, but also from milk fat globular membrane proteins, such as glycosylation-dependent cell adhesion molecule 1. RM and UHT milk showed comparable unmodified peptide profiles, whereas IF differed mainly due to a higher number of β-casein peptides. When 26 non-enzymatic posttranslational modifications (PTMs) were targeted in the milk peptidomes, 175 modified peptides were identified, i.e., mostly lactosylated and a few hexosylated or oxidized peptides. Most modified peptides originated from αS-caseins. The numbers of lactosylated peptides increased with harsher processing.


Splice-site mutations: a novel genetic mechanism of Crigler-Najjar syndrome type 1.
Crigler-Najjar syndrome type 1 (CN-1) is a recessively inherited, potentially lethal disorder characterized by severe unconjugated hyperbilirubinemia resulting from deficiency of the hepatic enzyme bilirubin-UDP-glucuronosyltransferase. In all CN-1 patients studied, structural mutations in one of the five exons of the gene (UGT1A1) encoding the uridinediphosphoglucuronate glucuronosyltransferase (UGT) isoform bilirubin-UGT1 were implicated in the absence or inactivation of the enzyme. We report two patients in whom CN-1 is caused, instead, by mutations in the noncoding intronic region of the UGT1A1 gene. One patient (A) was homozygous for a G-->C mutation at the splice-donor site in the intron, between exon 1 and exon 2. The other patient (B) was heterozygous for an A-->G shift at the splice-acceptor site in intron 3, and in the second allele a premature translation-termination codon in exon 1 was identified. Bilirubin-UGT1 mRNA is difficult to obtain, since it is expressed in the liver only. To determine the effects of these splice-junction mutations, we amplified genomic DNA of the relevant splice junctions. The amplicons were expressed in COS-7 cells, and the expressed mRNAs were analyzed. In both cases, splice-site mutations led to the use of cryptic splice sites, with consequent deletions in the processed mRNA. This is the first report of intronic mutations causing CN-1 and of the determination of the consequences of these mutations on mRNA structure, by ex vivo expression.


A cost effectiveness analysis of treatment options for methotrexate-naive rheumatoid arthritis.
New treatment options for patients with methotrexate (MTX)-naive rheumatoid arthritis (RA) have become available. Given wide variability in efficacy and cost among different treatment options, we sought to determine their relative cost effectiveness to help guide policy in different cost constrained settings. We performed a cost effectiveness analysis comparing 5 monotherapy options for patients with MTX-naive RA: (1) etanercept, (2) leflunomide, (3) MTX (up to 15 mg weekly), (4) sulfasalazine (SSZ), and (5) no second line agent. A decision analysis model was used with a time horizon of 6 months. We employed 2 measures of effectiveness based on published clinical trial data: American College of Rheumatology (ACR) 20% response proportion (ACR 20) and a weighted average of proportions achieving ACR 70, ACR 50, and ACR 20 (ACR 70 weighted response, ACR 70WR). Incremental cost effectiveness ratios were calculated as additional cost per patient achieving either outcome, compared with the next most expensive option. In both base case analyses employing ACR 20 and ACR 70WR as effectiveness measures, MTX and SSZ both cost less and were more effective (i.e., cost saving) than no second line agent. Leflunomide cost more and was less efficacious than SSZ (dominated) in analyses using either outcome. The most efficacious option, etanercept, cost US $41,900 per ACR 20 and $40,800 per ACR 70 WR compared with SSZ and MTX, respectively. When we included only direct costs in analyses, the least expensive non-dominated option was SSZ with incremental cost effectiveness ratios of US $900 per ACR 20 and $1500 per ACR 70WR compared with no second line agent. Overall, relative cost effectiveness between MTX and SSZ was sensitive to variation in relevant variables in sensitivity analyses. Otherwise, our extensive sensitivity analyses did not substantially affect the base case results. MTX is cost effective (cost saving vs the no second line agent option) for MTX-naive RA in achieving ACR 20 or ACR 70WR over a 6 month period. Based on available data, the relative cost effectiveness between SSZ and MTX cannot be determined with reasonable certainty and SSZ therapy appears to be as cost effective as MTX (cost saving) in achieving ACR outcomes over a 6 month period. The most efficacious option, etanercept, incurs much higher incremental costs per ACR 20 or ACR 70WR than other options analyzed. Whether etanercept compared with MTX is cost effective depends on whether > $40,000 per ACR 20 or ACR 70WR over a 6 month period is considered acceptable.


SU-E-T-444: 4D-CT and Active Breathing Coordinator Play Similar Role in Sparing Normal Liver Tissue in the Radiotherapy of Hepatocellular Carcinoma.
To investigate the difference in sparing normal liver tissue between 4D-CT and active breathing coordinator (ABC) in the radiotherapy of Hepatocellular carcinoma(HCC) applying RapidArc. Ten patients with HCC underwent 3D-CT at end inspiration hold(EIH) associated with active breathing coordinator(ABC),and 4D-CT scan. The 4D-CT images were sorted in to 10 serious CT images according respiratory phase. Gross tumor volumes(GTVs) were manually contoured on different CT images. The individual internal gross target volume(IGTV) was obtained from 10 GTVs of 4D-CT images. Plan target volumes (PTVs) were obtained based on IGTV and GTVEIH. PTV1 was obtained from IGTV plus 5 mm margins isotropically, and PTV2 was obtained from GTVEIH using 8 mm margins isotropically. RapidArc plans were designed on PTV1 and PTV2, for PTV1 RapidArc plans with one single whole arc were applied and for PTV2 three 135°arcs, and name RapidArc1 and RapidArc2. Dosimetric differences were compared. There were no significant difference in conformity index (0.93±0.05 Vs 0.93±0.03), homogeneity index(1.08±0.01 Vs 1.08±0.01), the Dl%(54.44±0.36 Gy Vs54.68±0.47 Gy) and D99%(49.18±0.48Gy Vs 49.00±0.34 Gy) between RapidArc1 and RapidArc2(P>0.05). There were no significant differences in the mean dose(8.23±1.5Gy Vs 7.63±3.00Gy),V5(46.64±19.31Vs43.29±19.94), V10(28.73±11.54 Vs 28.23±12.37), V15(17.87±6.61 Vs 18.20±7.55), V20(11.62±4.39 Vs 11.83±4.58), V25(7.71±2.91Vs 7.58±2.79),V30(5.14±2.01Vs5.05±1.79), 35(3.41±1.35Vs 3.35±1.21) and V40(2.12±0.86Vs2.06±0.75) between RapidArc1 and RapidArc2 (P>0.05). The RapidArc plans with three 135°arcs associated could achieve similar dose delivery as the plans with single whole arc, and 4D-CT and ABC could play similar role in sparing normal liver tissue assuring the accuracy of target volume in radiotherapy of HCC.


Excess heparin dosing among fibrinolytic-treated patients with ST-segment elevation myocardial infarction.
Although the use of heparin with fibrinolytics is associated with more rapid ST-segment resolution and increased infarct-related artery patency among patients with ST-segment elevation myocardial infarction (STEMI), its associated increase in bleeding risk is well documented and might be augmented by excess heparin dosing. We sought to characterize the incidence and associated bleeding risk of excess heparin dosing among patients with STEMI treated with fibrinolysis who were enrolled in the Can Rapid Risk Stratification of Unstable Angina Patients Suppress Adverse Outcomes with Early Implementation of the American College of Cardiology/American Heart Association Guidelines initiative. Excess dosing was defined as a bolus more than 60 U/kg or an infusion more than 12 U/kg/h per American College of Cardiology/American Heart Association guidelines and was further stratified into major and mild excess (major defined as a bolus>70 U/kg or infusion >15 U/kg/h). Among 964 fibrinolytic-treated patients with STEMI, 758 (79%) received adjunctive unfractionated heparin therapy. Of these, 368 patients (49%) received excess dosing of unfractionated heparin and 137 patients (18%) received major excess heparin dosing. Factors significantly associated with excess dosing included low body weight and female sex. Patients who received major excess dosing had higher unadjusted rates of major bleeding (19.2% vs 12.4%, P=.004) and transfusion (13.5% vs 4.7%, P=.0002) than patients without excess dosing. After adjustment, a trend persisted for the association with higher transfusion risk (odds ratio 1.39 [0.61-3.14]). Approximately half of fibrinolytic-treated patients with STEMI in contemporary practice received an excess dose of unfractionated heparin. Careful attention to dosing is needed to limit the compounded bleeding risk when heparin is added to fibrinolytic therapy.


Methylovirgula ligni gen. nov., sp. nov., an obligately acidophilic, facultatively methylotrophic bacterium with a highly divergent mxaF gene.
Two strains of Gram-negative, aerobic, non-pigmented, non-motile, rod-shaped bacteria were isolated from beechwood blocks during decay by the white-rot fungus Hypholoma fasciculare and were designated strains BW863(T) and BW872. They are capable of methylotrophic growth and assimilate carbon via the ribulose-bisphosphate pathway. In addition to methanol, the novel isolates utilized ethanol, pyruvate and malate. Strains BW863(T) and BW872 are obligately acidophilic, mesophilic organisms capable of growth at pH 3.1-6.5 (with an optimum at pH 4.5-5.0) and at 4-30 degrees C. Phospholipid fatty acid profiles of these bacteria contain unusually large amounts (about 90 %) of C(18 : 1)omega7c, thereby resembling the profiles of Methylobacterium strains. The predominant quinone is Q-10. The DNA G+C content of the novel isolates is 61.8-62.8 mol%. On the basis of 16S rRNA gene sequence similarity, strains BW863(T) and BW872 are most closely related to the acidophilic methanotroph Methylocapsa acidiphila B2(T) (96.5-97 %). Comparative sequence analysis of mxaF, the gene encoding the large subunit of methanol dehydrogenase, placed the MxaF sequences of the two novel strains in a cluster that is distinct from all previously described MxaF sequences of cultivated methylotrophs. The identity between the MxaF sequences of the acidophilic isolates and those from known alpha-, beta- and gammaproteobacterial methylotrophs was respectively 69-75, 61-63 and 64-67 %. The data therefore suggest that strains BW863(T) and BW872 represent a novel genus and species of methylotrophic bacteria, for which the name Methylovirgula ligni gen. nov., sp. nov. is proposed. Strain BW863(T) (=DSM 19998(T) =NCIMB 14408(T)) is the type strain of Methylovirgula ligni.


Development and characterization of a Yucatan miniature biomedical pig permanent middle cerebral artery occlusion stroke model.
Efforts to develop stroke treatments have met with limited success despite an intense need to produce novel treatments. The failed translation of many of these therapies in clinical trials has lead to a close examination of the therapeutic development process. One of the major factors believed to be limiting effective screening of these treatments is the absence of an animal model more predictive of human responses to treatments. The pig may potentially fill this gap with a gyrencephalic brain that is larger in size with a more similar gray-white matter composition to humans than traditional stroke animal models. In this study we develop and characterize a novel pig middle cerebral artery occlusion (MCAO) ischemic stroke model. Eleven male pigs underwent MCAO surgery with the first 4 landrace pigs utilized to optimize stroke procedure and 7 additional Yucatan stroked pigs studied over a 90 day period. MRI analysis was done at 24 hrs and 90 days and included T2w, T2w FLAIR, T1w FLAIR and DWI sequences and associated ADC maps. Pigs were sacrificed at 90 days and underwent gross and microscopic histological evaluation. Significance in quantitative changes was determined by two-way analysis of variance and post-hoc Tukey's Pair-Wise comparisons. MRI analysis of animals that underwent MCAO surgery at 24 hrs had hyperintense regions in T2w and DWI images with corresponding ADC maps having hypointense regions indicating cytotoxic edema consistent with an ischemic stroke. At 90 days, region of interest analysis of T1 FLAIR and ADC maps had an average lesion size of 59.17 cc, a loss of 8% brain matter. Histological examination of pig brains showed atrophy and loss of tissue, consistent with MRI, as well as glial scar formation and macrophage infiltration. The MCAO procedure led to significant and consistent strokes with high survivability. These results suggest that the pig model is potentially a robust system for the study of stroke pathophysiology and potential diagnostics and therapeutics.


Origami-like unfolding of hydro-actuated ice plant seed capsules.
Actuated plant materials are a source of inspiration for the design of adaptive materials and structures that are responsive to specific external stimuli. Hydro-responsive, metabolism-independent plant movements are particularly fascinating, because the extracted concepts are more amenable to transfer into engineering than those dependent on cellular activity. Here we investigate the structural and compositional basis of a sophisticated plant movement mechanism--the hydration-dependent unfolding of ice plant seed capsules. This reversible origami-like folding pattern proceeds via a cooperative flexing-and-packing mechanism actuated by a swellable cellulose layer filling specialized plant cells. Swelling is translated into a bidirectional organ movement through simple geometric constraints embedded in the hierarchical architecture of the ice plant valves. Extracted principles from this reliable and reversible actuated movement have relevance to the emerging field of 'programmable matter' with applications as far-reaching as the design of satellites and artificial muscles.


Addressing Trauma in Adaptive Physical Activity: A Call to Reflection and Action.
Trauma is pervasive, embodied, and can be perpetrated or perpetuated by researchers, educators, and practitioners, including those within adaptive physical activity (APA). In this article, we highlight the need to address trauma within APA as a matter of access and justice. We share various conceptualizations of trauma from psychiatric, embodied, anti-pathologizing, and sociopolitical perspectives. Trauma-informed practice is introduced as a framework for creating safer, more inclusive programs and services, so we can recognize the impacts of trauma and affirm those who experience it. As the first step to a multistep trauma-informed process, our aim is to raise awareness of trauma and introduce resources for enacting trauma-informed practice. We also pose difficult questions about how we, as "helping" practitioners, researchers, and educators may be perpetuating or perpetrating harm and trauma, in particular sanism, within our profession. Ultimately, we invite readers to join us in reflection and action toward anti-pathologizing trauma-informed APA.


Mapping spatio-temporal dynamics of single biomolecules in living cells.
We present a Bayesian framework for inferring spatio-temporal maps of diffusivity and potential fields from recorded trajectories of single molecules inside living cells. The framework naturally lets us regularise the high-dimensional inference problem using prior distributions in order to obtain robust results. To overcome the computational complexity of inferring thousands of map parameters from large single particle tracking datasets, we developed a stochastic optimisation method based on local mini-batches and parsimonious gradient calculation. We quantified the gain in convergence speed on numerical simulations, and we demonstrated for the first time temporal regularisation and aligned values of the inferred potential fields across multiple time segments. As a proof-of-concept, we mapped the dynamics of HIV-1 Gag proteins involved in the formation of virus-like particles (VLPs) on the plasma membrane of live T cells at high spatial and temporal resolutions. We focused on transient aggregation events lasting only on tenth of the time required for full VLP formation. The framework and optimisation methods are implemented in the TRamWAy open-source software platform for analysing single biomolecule dynamics.


Structure of Simian Immunodeficiency Virus Envelope Spikes Bound with CD4 and Monoclonal Antibody 36D5.
The human immunodeficiency virus type 1 (HIV-1)/simian immunodeficiency virus (SIV) envelope spike (Env) mediates viral entry into host cells. The V3 loop of the gp120 component of the Env trimer contributes to the coreceptor binding site and is a target for neutralizing antibodies. We used cryo-electron tomography to visualize the binding of CD4 and the V3 loop monoclonal antibody (MAb) 36D5 to gp120 of the SIV Env trimer. Our results show that 36D5 binds gp120 at the base of the V3 loop and suggest that the antibody exerts its neutralization effect by blocking the coreceptor binding site. The antibody does this without altering the dynamics of the spike motion between closed and open states when CD4 is bound. The interaction between 36D5 and SIV gp120 is similar to the interaction between some broadly neutralizing anti-V3 loop antibodies and HIV-1 gp120. Two conformations of gp120 bound with CD4 are revealed, suggesting an intrinsic dynamic nature of the liganded Env trimer. CD4 binding substantially increases the binding of 36D5 to gp120 in the intact Env trimer, consistent with CD4-induced changes in the conformation of gp120 and the antibody binding site. Binding by MAb 36D5 does not substantially alter the proportions of the two CD4-bound conformations. The position of MAb 36D5 at the V3 base changes little between conformations, indicating that the V3 base serves as a pivot point during the transition between these two states.IMPORTANCE Glycoprotein spikes on the surfaces of SIV and HIV are the sole targets available to the immune system for antibody neutralization. Spikes evade the immune system by a combination of a thick layer of polysaccharide on the surface (the glycan shield) and movement between spike domains that masks the epitope conformation. Using SIV virions whose spikes were "decorated" with the primary cellular receptor (CD4) and an antibody (36D5) at part of the coreceptor binding site, we visualized multiple conformations trapped by the rapid freezing step, which were separated using statistical analysis. Our results show that the CD4-induced conformational dynamics of the spike enhances binding of the antibody.


Hypertension with mineralocorticoid administration to the Long-Evans rat.
1. The Carworth Long-Evans rat has been reported to develop adrenal-regeneration hypertension but not deoxycorticosterone acetate (DOCA) hypertension. Deficiency of a hypothalamic receptor for deoxycorticosterone which mediates saline polydipsia has been postulated to underlie this resistance. Since a mineralocorticoid etiology for adrenal-regeneration hypertension has been postulated and all mineralocorticoids are thought to act on common receptors, these previous reports are difficult to reconcile. 2. To determine if an absolute or relative resistance to mineralocorticoids is present, Charles River Long-Evans and Sprague-Dawley rats were given 40 mg (107 micromol) of DOCA pellets/rat or 250 microgrms (0.65 micromol) of 2 alpha-methyl-9-alpha-fluorocortisol/day subcutaneously. 3. Saline polydipsia occurred with both steroids with both rat strains, though significantly less with the Long-Evans rats. Both types of rats became hypertensive and developed cardiac and renal enlargement with both steroids. Hypertension developed more rapidly with 2 alpha-methyl-9 alpha-fluorocortisol. 4. Thus mineralocorticoid hypertension can be produced in the Charles River Long-Evans rat, and the development of adrenal-regeneration hypertension in this rat strain is not incompatible with a mineralocorticoid etiology for adrenal-regeneration hypertension.


Control of Alzheimer's amyloid beta toxicity by the high molecular weight immunophilin FKBP52 and copper homeostasis in Drosophila.
FK506 binding proteins (FKBPs), also called immunophilins, are prolyl-isomerases (PPIases) that participate in a wide variety of cellular functions including hormone signaling and protein folding. Recent studies indicate that proteins that contain PPIase activity can also alter the processing of Alzheimer's Amyloid Precursor Protein (APP). Originally identified in hematopoietic cells, FKBP52 is much more abundantly expressed in neurons, including the hippocampus, frontal cortex, and basal ganglia. Given the fact that the high molecular weight immunophilin FKBP52 is highly expressed in CNS regions susceptible to Alzheimer's, we investigated its role in Abeta toxicity. Towards this goal, we generated Abeta transgenic Drosophila that harbor gain of function or loss of function mutations of FKBP52. FKBP52 overexpression reduced the toxicity of Abeta and increased lifespan in Abeta flies, whereas loss of function of FKBP52 exacerbated these Abeta phenotypes. Interestingly, the Abeta pathology was enhanced by mutations in the copper transporters Atox1, which interacts with FKBP52, and Ctr1A and was suppressed in FKBP52 mutant flies raised on a copper chelator diet. Using mammalian cultures, we show that FKBP52 (-/-) cells have increased intracellular copper and higher levels of Abeta. This effect is reversed by reconstitution of FKBP52. Finally, we also found that FKBP52 formed stable complexes with APP through its FK506 interacting domain. Taken together, these studies identify a novel role for FKBP52 in modulating toxicity of Abeta peptides.


Profile of Self-Reported Physical Tasks and Physical Training in Brazilian Special Operations Units: A Web-Based Cross-Sectional Study.
There is limited research examining the physical tasks that Brazilian special policemen groups can perform in the line of duty. The aims of this study were to (a) identify the occupational tasks of specialist police personnel serving in the Rapid Response Group (GRR) and Tactical Operations Command (COT), and to profile the frequency, difficulty, and importance of these tasks, and (b) to explore the current physical training these special operations police units (SOPUs) officers undertake to maintain their operational fitness. Univariate analysis for numerical variables (mean and standard deviation (SD)), as well as the absolute and relative frequencies for categorical variables were performed. Two Brazilian SOPUs performed a questionnaire with demographic, performance, and physical training sections. A total of 78 respondents (24 of the GRR and 54 of the COT) completed the survey. "Standing and/or sitting with complete equipment for long periods in different climatic conditions", and "lifting/pushing/pulling heavier objects" were the most frequent and difficult occupational tasks of both SOPUs, respectively. "Shooting a long weapon" and "breaking a door" were the most important for GRR and COT, respectively. All officers undertook regular physical training (~9 h/week), in an unstructured manner, without supervision, and planning of physical training is carried out autonomously (COT) or a mixture of autonomously and directed (GRR), with the main objectives of developing aerobic endurance and muscle strength. It is important that SOPUs teams train their members' physical capabilities to perform the tasks identified in this study, as well as follow a structured, supervised, and planned physical training program.


In-line particle holography with an astigmatic beam: setup self-calibration using an "inverse problems" approach.
The use of digital in-line holography for the characterization of confined flows in cylindrical geometry confinements (e.g., cylindrical pipe or cylindrical capillaries) is discussed. Due to cylindrical geometry of the walls, the illuminating laser wave can be strongly astigmatic, which renders the use of classical reconstruction techniques impossible. Contrary to plane wave holography setup, the diffraction pattern of the particles strongly depends on the axial distance of the latter to the entry face of the confinement structure. To address this reconstruction issue, we propose to use an "inverse problems" approach. This approach amounts to finding the best match (least squares solution) between a diffraction pattern model and the captured hologram. For this purpose, a direct imaging model for astigmatic holograms, based on the use of transfer matrices, is presented and validated by comparing experimental and simulated holograms. The accuracy of the "inverse problems" reconstruction is then used to calibrate the experimental setup adjustable parameters. Finally, the approach is tested through experimental astigmatic hologram reconstruction, thus paving the way to its use in pipe flow studies.


Functional domains of the human epididymal protease inhibitor, eppin.
Eppin has two potential protease inhibitory domains: a whey acid protein or four disulfide core domain and a Kunitz domain. The protein is also reported to have antibacterial activity against Gram-negative bacteria. Eppin and its whey acid protein and Kunitz domains were expressed in Escherichia coli and their ability to inhibit proteases and kill bacteria compared. The Kunitz domain inhibits elastase (EC 184.108.40.206) to a similar extent as intact eppin, whereas the whey acid protein domain has no such activity. None of these fragments inhibits trypsin (EC 220.127.116.11) or chymotrypsin (EC 184.108.40.206) at the concentrations tested. In a colony forming unit assay, both domains have some antibacterial activity against E. coli, but this was not to the same degree as intact eppin or the two domains together. When bacterial respiratory electron transport was measured using a 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide assay, eppin and its domains caused an increase in the rate of respiration. This suggests that the mechanism of cell killing may be partly through the permeablization of the bacterial inner membrane, resulting in uncoupling of respiratory electron transport and consequent collapse of the proton motive force. Thus, we conclude that although both of eppin's domains are involved in the protein's antibacterial activity, only the Kunitz domain is required for selective protease inhibition.


The independent relationship between trouble controlling Facebook use, time spent on the site and distress.
There is an emerging literature base on the relationship between maladaptive traits and "addiction" to social networking sites. These studies have operationalized addiction as either spending excessive amounts of time on social networking sites (SNS) or trouble controlling SNS use, but have not assessed the unique contribution of each of these constructs on outcomes in the same models. Moreover, these studies have exclusively been conducted with younger people rather than a heterogeneous sample. This study examined the independent relationship of a brief Facebook addiction scale, time spent on Facebook, and Facebook checking on positive and negative social domains, while controlling for self-esteem and social desirability. Participants were recruited using e-mail, SNS posts and through Amazon's MTurk system. The sample included 489 respondents ages from 18 to approximately 70, who completed a 10-15 minute survey. Results indicate that neither time spent on Facebook nor Facebook checking was significantly associated with either self-esteem, fear of negative social evaluation or social comparison, while SNS addiction symptoms were each independently associated with Facebook usage. Neither time spent on Facebook nor SNS addiction symptoms were associated with positive social relationships. Overall results suggest that time on SNS and trouble controlling use should be considered independent constructs and that interventions should target underlying loss of control as the primary intervention target above ego syntonic time spent on the site.


Therapy of hypoparathyroidism by replacement with parathyroid hormone.
Hypoparathyroidism (HypoPT) is a state of hypocalcemia due to inappropriate low levels of parathyroid hormone (PTH). HypoPT is normally treated by calcium supplements and activated vitamin D analogues. Although plasma calcium is normalized in response to conventional therapy, quality of life (QoL) seems impaired and patients are at increased risk of renal complications. A number of studies have suggested subcutaneous injections with PTH as an alternative therapy. By replacement with the missing hormone, urinary calcium may be lowered and QoL may improve. PTH replacement therapy (PTH-RT) possesses, nevertheless, a number of challenges. If PTH is injected only once a day, fluctuations in calcium levels may occur resulting in hypercalcemia in the hours following an injection. Twice-a-day injections seem to cause less fluctuation in plasma calcium but do stimulate bone turnover to above normal. Most recently, continuous delivery of PTH by pump has appeared as a feasible alternative to injections. Plasma calcium levels do not fluctuate, urinary calcium is lowered, and bone turnover is only stimulated modestly (into the normal range). Further studies are needed to assess the long-term effects. If beneficial, it seems likely that standard treatment of HypoPT in the future will change into replacement therapy with the missing hormone.


Early diagnosis of cutaneous melanoma: an observation in southern Brazil.
The incidence of melanoma cutaneous and the mortalities rates are rising in most countries worldwide. to describe the histopathological characteristics of cutaneous melanoma, according to the criteria established by the Brazilian Group of Melanoma, and to evaluate early diagnosis in a cancer treatment referral center. we performed a cross-sectional descriptive study of cases of primary cutaneous melanoma identified after excisional biopsy and processed at the pathology laboratory of Complexo Hospitalar Santa Casa between Jan 1st 2000 and Jan 15th 2005. The following variables were analyzed: age, gender, topography, histopathologic subtype, Breslow thickness, growth phase, Clark level, mitotic index, peritumoral and intratumoral lymphocytic inflammatory infiltrate, angiolymphatic invasion, ulceration, regression, type of regression, microscopic satellitosis, and surgical margins. 328 cases, 57% female and 43% male, were analyzed. Mean age was 55.6 years. For women, the most common tumor location was in inferior(29.26%) and superior limbs(23.94%), while for men melanoma was mainly found in the back(35%), followed by anterior chest/abdomen(14.29%) (p<0.05). Prevalence of histologic subtypes was the following: superficial spreading melanoma(62.8%), lentigo maligna(14.9%), nodular(14.6%), acral(7.3%), and desmoplastic(0.3%) types. Regarding Breslow, 26.2% were in situ, 36.9% had <1 mm, and only 15.9% were ? 4mm in depth. the distribution of histopathologic subtypes, as well as Breslow thickness, was in accordance with previous studies in outpatient populations. The profile of cases of cutaneous melanoma diagnosed in a tertiary hospital seems to be experiencing some changes over the last two decades, with a current trend for earlier diagnosis.


Control of mouse cardiac morphogenesis and myogenesis by transcription factor MEF2C.
Members of the myocyte enhancer factor-2 (MEF2) family of MADS (MCM1, agamous, deficiens, serum response factor)-box transcription factors bind an A-T-rich DNA sequence associated with muscle-specific genes. The murine MEF2C gene is expressed in heart precursor cells before formation of the linear heart tube. In mice homozygous for a null mutation of MEF2C, the heart tube did not undergo looping morphogenesis, the future right ventricle did not form, and a subset of cardiac muscle genes was not expressed. The absence of the right ventricular region of the mutant heart correlated with down-regulation of the dHAND gene, which encodes a basic helix-loop-helix transcription factor required for cardiac morphogenesis. Thus, MEF2C is an essential regulator of cardiac myogenesis and right ventricular development.


Is cooperation viable in mobile organisms? Simple Walk Away rule favors the evolution of cooperation in groups.
The evolution of cooperation through partner choice mechanisms is often thought to involve relatively complex cognitive abilities. Using agent-based simulations I model a simple partner choice rule, the 'Walk Away' rule, where individuals stay in groups that provide higher returns (by virtue of having more cooperators), and 'Walk Away' from groups providing low returns. Implementing this conditional movement rule in a public goods game leads to a number of interesting findings: 1) cooperators have a selective advantage when thresholds are high, corresponding to low tolerance for defectors, 2) high thresholds lead to high initial rates of movement and low final rates of movement (after selection), and 3) as cooperation is selected, the population undergoes a spatial transition from high migration (and a many small and ephemeral groups) to low migration (and large and stable groups). These results suggest that the very simple 'Walk Away' rule of leaving uncooperative groups can favor the evolution of cooperation, and that cooperation can evolve in populations in which individuals are able to move in response to local social conditions. A diverse array of organisms are able to leave degraded physical or social environments. The ubiquitous nature of conditional movement suggests that 'Walk Away' dynamics may play an important role in the evolution of social behavior in both cognitively complex and cognitively simple organisms.


Slow-onset, long-duration, alkyl analogues of methylphenidate with enhanced selectivity for the dopamine transporter.
Methylphenidate analogues, in which the carbomethoxy has been replaced by an alkyl group and with different phenyl substituents, have been synthesized and tested in monoamine transporter assays. As predicted from a pharmacophore model, most of the RR/SS diastereomers showed high potency as dopamine reuptake inhibitors. Analogues with a 4-chlorophenyl group and an unbranched initial alkyl atom had consistently enhanced selectivity for the dopamine transporter. The most potent compounds were those with a three- or four-carbon chain. The "inactive" RS/SR diastereomers showed substantial activity when the phenyl substituent was 3,4-dichloro. On a locomotor assay, one compound was found to have a slow onset and a long duration of action. The activity of these compounds provides additional evidence for a conformational/superposition model of methylphenidate with cocaine-like structures. A ketone analogue, obtained by hydrogenating a previously described vinylogous amide, had activity similar to that of methylphenidate.


Analysis of postural perturbation responses.
People with cerebellar ataxia lack lower limb coordination and dissipate sway motion slowly and inefficiently after a posture perturbation. We report a practical and low-cost "human resonance frequency test" for both laboratory and clinical use to quantify progress in balance and cerebellar rehabilitation. We assumed that the center-of-pressure (COP) oscillation rate of decay following a standing posture perturbation is directly related to resonance frequency; a more rapidly dissipating COP oscillation about the position of equilibrium indicates, by definition, more efficient postural control. We hypothesized that following successful physical rehabilitation, people with cerebellar degeneration will have a faster rate of decay of the COP response to an external perturbation. Because the COP is modulated by a synergy of trunk and lower limb motion strategies, COP decay rate may be a useful measure of lower limb coordination in people with cerebellar ataxia. The method was applied to three subjects with cerebellar ataxia before and after rehabilitation; there was good agreement between the calculated COP decay rate and conventionally used gait stability parameters providing pilot data for this simple approach.


Spermatogenesis in the testes of diapause and non-diapause pupae of the sweet potato hornworm, Agrius convolvuli (L.) (Lepidoptera: Sphingidae).
Dichotomous spermatogenesis was examined in relation to diapause in the sweet potato hornworm, Agrius convolvuli. In non-diapause individuals, eupyrene metaphase began during the fifth larval instar and eupyrene spermatids appeared in wandering larvae. Bundles of mature sperm were found after pupation. Apyrene spermatocytes also appeared during the fifth larval instar, but meiotic divisions occurred irregularly and their nuclei were discarded from the cells during spermiogenesis. Morphometric analyses of flagellar axonemes showed a variable sperm number in apyrene bundles. The variation ranging from 125 to 256 sperm per bundle indicated abnormal divisions or the elimination of apyrene spermatocytes. In diapause-induced hornworms, spermatogenesis progressed similarly during the larval stages. The cessation of spermatogenesis during diapause is characterized by 1) secondary spermatocytes and sperm bundles degenerating gradually as the diapause period lengthens, and 2) spermatogonia or primary spermatocytes appearing throughout diapause. A TUNEL (TdT-mediated dUTP-biotin nick end-labeling) assay revealed that DNA fragmentation occurred in the nuclei of secondary spermatocytes and early spermatids. Aggregates of heterochromatin along the nuclear membrane indicated the onset of apoptosis, and condensed chromatin was confirmed by electron microscopy to be the apoptotic body. These results show that the degenerative changes in spermatogenic cells during pupal diapause were controlled by apoptosis.


Guillain-Barré syndrome in Colombia: where do we stand now?
Guillain-Barré syndrome (GBS) is a rapid-onset muscle weakness disease caused by the immune-mediated damage of the peripheral nervous system. Since there is an increase incidence of GBS cases in Latin America, particularly in Colombia, and most of them are currently preceded by Zika virus (ZIKV) infection, we aimed to assess the available evidence of the disease in Colombia through a systematic literature review. Out of 51 screened abstracts, only 16 corresponded to articles that met inclusion criteria, of which 15 were case reports or case series. A total of 796 cases of GBS were reported in the included articles. The majority of patients were males (66.8 %) and younger than 50 years old (94 %). An infectious disease before the onset of GBS was registered in 31 % of patients, with gastrointestinal or respiratory symptoms being the most frequently observed. In those cases in which electrodiagnostic tests were performed, the most common subphenotype was acute inflammatory demyelinating polyneuropathy (17 %). Death was reported in 15 % of patients. Data regarding GBS in Colombia is scant and heterogeneous. Taking into account the burden of the disease and the recent rise of GBS cases associated with ZIKV, a careful patient evaluation and a systematic collection of data are warranted. A form to data gathering is proposed.


Human lysyl-tRNA synthetase phosphorylation promotes HIV-1 proviral DNA transcription.
Human lysyl-tRNA synthetase (LysRS) was previously shown to be re-localized from its normal cytoplasmic location in a multi-aminoacyl-tRNA synthetase complex (MSC) to the nucleus of HIV-1 infected cells. Nuclear localization depends on S207 phosphorylation but the nuclear function of pS207-LysRS in the HIV-1 lifecycle is unknown. Here, we show that HIV-1 replication was severely reduced in a S207A-LysRS knock-in cell line generated by CRISPR/Cas9; this effect was rescued by S207D-LysRS. LysRS phosphorylation up-regulated HIV-1 transcription, as did direct transfection of Ap4A, an upstream transcription factor 2 (USF2) activator that is synthesized by pS207-LysRS. Overexpressing an MSC-derived peptide known to stabilize LysRS MSC binding inhibited HIV-1 replication. Transcription of HIV-1 proviral DNA and other USF2 target genes was reduced in peptide-expressing cells. We propose that nuclear pS207-LysRS generates Ap4A, leading to activation of HIV-1 transcription. Our results suggest a new role for nuclear LysRS in facilitating HIV-1 replication and new avenues for antiviral therapy.


Updated method guidelines for cochrane musculoskeletal group systematic reviews and metaanalyses.
The Cochrane Musculoskeletal Group (CMSG), one of 53 groups of the not-for-profit, international Cochrane Collaboration, prepares, maintains, and disseminates systematic reviews of treatments for musculoskeletal diseases. It is important that authors conducting CMSG reviews and the readers of our reviews be aware of and use updated, state-of-the-art systematic review methodology. One hundred sixty reviews have been published. Previous method guidelines for systematic reviews of interventions in the musculoskeletal field published in 2006 have been substantially updated to incorporate methodological advances that are mandatory or highly desirable in Cochrane reviews and knowledge translation advances. The methodological advances include new guidance on searching, new risk-of-bias assessment, grading the quality of the evidence, the new Summary of Findings table, and comparative effectiveness using network metaanalysis. Method guidelines specific to musculoskeletal disorders are provided by CMSG editors for various aspects of undertaking a systematic review. These method guidelines will help improve the quality of reporting and ensure high standards of conduct as well as consistency across CMSG reviews.


In vitro and in vivo modification of Neisseria gonorrhoeae lipooligosaccharide epitope structure by sialylation.
After growth of gonococci in the presence of cytidine monophospho-N-acetyl-neuraminic acid (CMP-NANA), their 4.5-kD lipooligosaccharide (LOS) component was increased by approximately 400 daltons, whereas the LOS of strains lacking the 4.5-kD component were unaffected. Expression of mAb-defined epitopes on the 4.5-kD component was decreased on LOS of strains grown in CMP-NANA, and treatment of the LOS with neuraminidase reversed this affect. Gonococci incubated with human PMNs also had decreased expression of the 4.5-kD+ epitopes. A detergent extract of gonococci incorporated radiolabeled NANA in the LOS, suggesting the presence of a sialyltransferase in gonococci. Exogenous sialyltransferases also could use LOS as an acceptor.