Where the eye looks, the hand follows; limb-dependent magnetic misreaching in optic ataxia.
The posterior parietal cortex (PPC) is thought to play an important role in the sensorimotor transformations associated with reaching movements. In humans, damage to the PPC, particularly bilateral lesions, leads to impairments of visually guided reaching movements (optic ataxia). Recent accounts of optic ataxia based upon electrophysiological recordings in monkeys have proposed that this disorder arises because of a breakdown in the tuning fields of parietal neurons responsible for integrating spatially congruent retinal, eye, and hand position signals to produce coordinated eye and hand movements . We present neurological evidence that forces a reconceptualization of this view. We report a detailed case study of a patient with a limb-dependent form of optic ataxia who can accurately reach with either hand to objects that he can foveate (thereby demonstrating coordinated eye-hand movements) but who cannot effectively decouple reach direction from gaze direction for movements executed using his right arm. The demonstration that our patient's misreaching is confined to movements executed using his right limb, and only for movements that are directed to nonfoveal targets, rules out explanations based upon simple perceptual or motor deficits but indicates an impairment in the ability to dissociate the eye and limb visuomotor systems when appropriate.


Respiratory health effects of the fiberglass-reinforced plastic lamination process in the yacht-building industry.
Objectives Fiberglass-reinforced plastics (FRP) manufacturing has been related to cases of severe airway obstruction and elevated risk of respiratory mortality. But the specific job content risk is not clear. This study evaluated the respiratory health effects of the FRP lamination process. Methods A questionnaire was used to evaluate respiratory symptoms of workers in two yacht-building plants. Pre-shift (07:30-08:30 hours) and post-shift (17:00-18:00 hours) lung function was measured, while post-shift induced sputum was collected on the first day of the week. The participants were grouped into FRP laminators and non-laminators. Linear and logistic regression was used to investigate the effects of the lamination process on lung function. Results Laminators had a higher prevalence of chronic cough, lower pre-shift forced expiratory volume in first second (FEV1) and FEV1/force vital capacity (FVC) (-3.3% and -1.5%), lower post-shift FVC and FEV1 (-3.6% and -4.9%), and larger post-shift reduction of FVC (-2.1%) compared to non-laminators. The laminators also had higher risk of early obstructive and overall (obstructive plus restrictive) lung function impairment, and post-shift reduction of FVC >10% [odds ratio (OR) 5.98, 4.98, and 3.87, respectively). They also had higher percentages of neutrophils and lymphocytes in the induced sputum. Conclusion Laminators should undergo regular check-ups of respiratory symptoms and lung function. Further toxicologic studies are warranted to identify the specific causal agent in the FRP lamination process.


Does this adult patient have early HIV infection?: The Rational Clinical Examination systematic review.
Timely identification of human immunodeficiency virus (HIV) infection in adults can contribute to reduced mortality and likelihood of further HIV transmission. During the first 6 months after infection, known as early HIV infection, patients often report a well-described constellation of symptoms and signs. However, the literature examining utility of the clinical examination in identifying early infection has not been systematically assessed. To assess the accuracy of symptoms and signs in identifying early HIV infection among adults. We searched MEDLINE and EMBASE (1981-May, 2014) for articles investigating symptoms and signs of early HIV infection in adults and searched reference lists of retrieved articles. We retained original studies that compared symptoms and signs among patients with early HIV infection in comparison to HIV-negative individuals. Data were extracted and used to calculate sensitivity, specificity, and likelihood ratios (LRs), and meta-analysis was used to calculate summary LRs. Of 1356 studies, 16 studies included data that were eligible for meta-analysis and included a total of 24,745 patients and 1253 cases of early HIV infection. Symptoms that increased the likelihood of early HIV infection the most included genital ulcers (LR, 5.4; 95% CI, 2.5-12), weight loss (LR, 4.7; 95% CI, 2.1-7.2), vomiting (LR, 4.6; 95% CI, 2.5-8.0), and swollen lymph nodes (LR, 4.6; 95% CI, 1.3-8.0). No symptoms had an LR that was 0.5 or lower, but the absence of recent fever (LR, 0.74; 95% CI, 0.64-0.84) slightly decreased the likelihood of early HIV infection. The presence of lymphadenopathy on physical examination was the most useful sign (LR, 3.1; 95% CI, 1.0-5.2). No sign had an LR of 0.5 or less, but the absence of lymphadenopathy slightly decreased the likelihood of early HIV infection (LR, 0.70, 95% CI, 0.49-0.92). Using data from studies that considered combinations of findings (range of possible findings, 4-17), the summary LR for individuals with 0 findings was 0.47 (95% CI, 0.38-0.58). The limited utility of the clinical examination to detect or rule out early HIV infection highlights the importance of routine testing for HIV infection among adults.


ECVAM's collaborations with academia.
The encouragement of ECVAM's connections with academia through direct collaboration and co-sponsored studentships has resulted in the successful achievement of higher degree qualifications for the young participants, and the development and promotion of alternative methods. So far, 26 students have been registered for higher degrees, of which 13 have been awarded so far, and 16 university departments in nine European countries have been directly involved. When other collaborations are included, the number of ECVAM's interactions with academic institutions rises to 33 departments in eleven countries, including the USA. In addition, through contracts awarded to academic institutions and other forms of collaboration, the prevalidation and validation of alternative methods have been progressed.


Catastrophic flood and forest cover change in the Huong river basin, central Viet Nam: a gap between common perceptions and facts.
Recent catastrophic floods in Viet Nam have been increasingly linked to land use and forest cover change in the uplands. Despite the doubts that many scientists have expressed on such nexus, this common view prompted both positive forest protection/reforestation programs and often-unwarranted blame on upland communities for their forest management practices. This study discusses the disparity between public perceptions and scientific evidences relating the causes of catastrophic floods. The former was drawn on the results of a questionnaire and focus groups discussions with key informants of different mountainous communities, whereas the latter was based on GIS and remote sensing analysis of land cover change, including a statistical analysis of hydro-meteorological data of the Huong river basin in Viet Nam. Results indicate that there is a gap between the common beliefs and the actual relationship between the forest cover change and catastrophic floods. Undeniably, the studied areas showed significant changes in land cover over the period 1989-2008, yet, 71% of the variance of catastrophic flood level in the downstream areas appeared related to variance in rainfall. Evidences from this study showed that the overall increasing trends of catastrophic flooding in the Huong river basin was mainly due to climate variability and to the development of main roads and dyke infrastructures in the lowlands. Forest management policies and programs, shaped on the common assumption that forest degradation in the upland is the main cause of catastrophic flood in the downstream areas, should be reassessed to avoid unnecessary strain on upland people.


Structural investigation of disordered stress proteins. Comparison of full-length dehydrins with isolated peptides of their conserved segments.
Dehydrins constitute a class of intrinsically disordered proteins that are expressed under conditions of water-related stress. Characteristic of the dehydrins are some highly conserved stretches of seven to 17 residues that are repetitively scattered in their sequences, the K-, S-, Y-, and Lys-rich segments. In this study, we investigate the putative role of these segments in promoting structure. The analysis is based on comparative analysis of four full-length dehydrins from Arabidopsis (Arabidopsis thaliana; Cor47, Lti29, Lti30, and Rab18) and isolated peptide mimics of the K-, Y-, and Lys-rich segments. In physiological buffer, the circular dichroism spectra of the full-length dehydrins reveal overall disordered structures with a variable content of poly-Pro helices, a type of elongated secondary structure relying on bridging water molecules. Similar disordered structures are observed for the isolated peptides of the conserved segments. Interestingly, neither the full-length dehydrins nor their conserved segments are able to adopt specific structure in response to altered temperature, one of the factors that regulate their expression in vivo. There is also no structural response to the addition of metal ions, increased protein concentration, or the protein-stabilizing salt Na(2)SO(4). Taken together, these observations indicate that the dehydrins are not in equilibrium with high-energy folded structures. The result suggests that the dehydrins are highly evolved proteins, selected to maintain high configurational flexibility and to resist unspecific collapse and aggregation. The role of the conserved segments is thus not to promote tertiary structure, but to exert their biological function more locally upon interaction with specific biological targets, for example, by acting as beads on a string for specific recognition, interaction with membranes, or intermolecular scaffolding. In this perspective, it is notable that the Lys-rich segment in Cor47 and Lti29 shows sequence similarity with the animal chaperone HSP90.


Kinotypes: stable species- and individual-specific profiles of cellular kinase activity.
Recently, questions have been raised regarding the ability of animal models to recapitulate human disease at the molecular level. It has also been demonstrated that cellular kinases, individually or as a collective unit (the kinome), play critical roles in regulating complex biology. Despite the intimate relationship between kinases and health, little is known about the variability, consistency and stability of kinome profiles across species and individuals. As a preliminary investigation of the existence of species- and individual-specific kinotypes (kinome signatures), peptide arrays were employed for the analysis of peripheral blood mononuclear cells collected weekly from human and porcine subjects (n = 6) over a one month period. The data revealed strong evidence for species-specific signalling profiles. Both humans and pigs also exhibited evidence for individual-specific kinome profiles that were independent of natural changes in blood cell populations. Species-specific kinotypes could have applications in disease research by facilitating the selection of appropriate animal models or by revealing a baseline kinomic signature to which treatment-induced profiles could be compared. Similarly, individual-specific kinotypes could have implications in personalized medicine, where the identification of molecular patterns or signatures within the kinome may depend on both the levels of kinome diversity and temporal stability across individuals.


Amphioxus and ascidian Dmbx homeobox genes give clues to the vertebrate origins of midbrain development.
The ancestral chordate neural tube had a tripartite structure, comprising anterior, midbrain-hindbrain boundary (MHB) and posterior regions. The most anterior region encompasses both forebrain and midbrain in vertebrates. It is not clear when or how the distinction between these two functionally and developmentally distinct regions arose in evolution. Recently, we reported a mouse PRD-class homeobox gene, Dmbx1, expressed in the presumptive midbrain at early developmental stages, and the hindbrain at later stages, with exclusion from the MHB. This gene provides a route to investigate the evolution of midbrain development. We report the cloning, genomic structure, phylogeny and embryonic expression of Dmbx genes from amphioxus and from Ciona, representing the two most closely related lineages to the vertebrates. Our analyses show that Dmbx genes form a distinct, ancient, homeobox gene family, with highly conserved sequence and genomic organisation, albeit more divergent in Ciona. In amphioxus, no Dmbx expression is observed in the neural tube, supporting previous arguments that the MHB equivalent region has been secondarily modified in evolution. In Ciona, the CiDmbx gene is detected in neural cells caudal to Pax2/5/8-positive cells (MHB homologue), in the Hox-positive region, but, interestingly, not in any cells rostral to them. These results suggest that a midbrain homologue is missing in Ciona, and argue that midbrain development is a novelty that evolved specifically on the vertebrate lineage. We discuss the evolution of midbrain development in relation to the ancestry of the tripartite neural ground plan and the origin of the MHB organiser.


Is the Podarcis muralis lizard left-eye lateralised when exploring a new environment?
The typical lateral eye position in ectotherms likely facilitated the spread of visual lateralisation--i.e., the different use of the eyes--in those species. The diffusion of this form of lateralisation seems due to the possibility of carrying out more than one task simultaneously, some controlled by one eye and the visual structures it feeds and some by the other. Similar to other species, exploratory and monitoring behaviours seem to be under left "eye system" control. Wild individuals of the Common wall lizard Podarcis muralis were tested individually in captivity to ascertain whether they showed lateralisation when exploring a new environment, using preferentially the left eye. In Experiment 1, the lizards explored a maze. A left-turning bias was found, both at individual and population level, indicating a possible right hemisphere visual control. In Experiment 2, lizards explored a T-maze, preferring to enter the left rather than the right arm though without any particular preference in the head turns. In Experiment 3, the lizards had to exit an opaque box within a terrarium. We found a left-eye preference again for head turn while leaving the box. Our findings support the hypothesis of right hemisphere mediation of exploratory and monitoring behaviours in Podarcis muralis. In addition to previous studies on the same species, our results support the hypothesis of a simultaneous control of anti-predatory and exploratory behaviours (left-eye mediated) and predatory behaviour (right-eye mediated).


Synthetic circuit designs for earth terraformation.
Mounting evidence indicates that our planet might experience runaway effects associated to rising temperatures and ecosystem overexploitation, leading to catastrophic shifts on short time scales. Remediation scenarios capable of counterbalancing these effects involve geoengineering, sustainable practices and carbon sequestration, among others. None of these scenarios seems powerful enough to achieve the desired restoration of safe boundaries. We hypothesize that synthetic organisms with the appropriate engineering design could be used to safely prevent declines in some stressed ecosystems and help improving carbon sequestration. Such schemes would include engineering mutualistic dependencies preventing undesired evolutionary processes. We hypothesize that some particular design principles introduce unescapable constraints to the engineered organisms that act as effective firewalls. Testing this designed organisms can be achieved by using controlled bioreactor models, with single and heterogeneous populations, and accurate computational models including different scales (from genetic constructs and metabolic pathways to population dynamics). Our hypothesis heads towards a future anthropogenic action that should effectively act as Terraforming processes. It also implies a major challenge in the existing biosafety policies, since we suggest release of modified organisms as potentially necessary strategy for success.


Three-year outcomes of the postapproval study of the AtriCure Bipolar Radiofrequency Ablation of Permanent Atrial Fibrillation Trial.
The Cox Maze IV operation is commonly performed concomitant with other cardiac operations and effectively reduces the burden of atrial fibrillation. Prospective randomized trials have reported outcomes early and at 12 months, but only single-center late durability results are available. As part of the postapproval process for a bipolar radiofrequency ablation system, we sought to determine early and midterm outcomes of patients undergoing the Cox Maze IV operation. A prospective, multicenter, single-arm study of 363 patients (mean age, 70 years, 82% valve surgery) with nonparoxysmal atrial fibrillation (mean duration, 60 months, 94% Congestive heart failure, Hypertension, Age ≥ 75, Diabetes, Stroke, VAScular disease, Age 65-74, Sex category ≥2) undergoing concomitant Maze IV atrial fibrillation ablation at 40 sites with 70 surgeons was performed between June 2010 and October 2014. Compliance with the study lesion set was 94.5%, and 99% had left atrial appendage closure. Freedom from atrial fibrillation was determined by extended monitoring, with a 48-hour Holter monitor minimum. There were no device-related complications. Freedom from atrial fibrillation off antiarrhythmic medications at 1, 2, and 3 years was 66%, 65%, and 64%, respectively, and including those using antiarrhythmics was 80%, 78%, and 76%, respectively. Warfarin was used in 49%, 44%, and 40%, respectively. In patients with nonparoxysmal atrial fibrillation, compliance with the protocol was high, and freedom from atrial fibrillation off antiarrhythmics was high and sustained to 3 years. The safety and effectiveness of the system and Cox Maze IV procedure support the Class I guideline recommendation for concomitant atrial fibrillation ablation in patients undergoing cardiac surgery.


Directional functions for orientation distribution estimation.
Computing the orientation distribution function (ODF) from high angular resolution diffusion imaging (HARDI) signals makes it possible to determine the orientation of fiber bundles of the brain. The HARDI signals are samples measured from a spherical shell and thus require processing on the sphere. Past work on ODF estimation involved using the spherical harmonics or spherical radial basis functions. In this work, we propose three novel directional functions able to represent the measured signals in a very compact manner, i.e., they require very few parameters to completely describe the measured signal. Analytical expressions are derived for computing the corresponding ODF. The directional functions can represent diffusion in a particular direction and mixture models can be used to represent multi-fiber orientations. We show how to estimate the parameters of this mixture model and elaborate on the differences between these functions. We also compare this general framework with estimation of ODF using spherical harmonics on some real and synthetic data. The proposed method could be particularly useful in applications such as tractography and segmentation. Details are also given on different ways in which interpolation can be performed using directional functions. In particular, we discuss a complete Euclidean as well as a "hybrid" framework, comprising of the Riemannian as well as Euclidean spaces, to perform interpolation and compute geodesic distances between two ODF's.


Membrane potential oscillations in molluscan "burster" neurones.
Membrane potential oscillations can be induced in molluscan neurones under a variety of artificial conditions. In the so-called 'burster' neurones oscillations are generated even in isolated cells. A likely mechanism for 'bursting' involves the following ionic currents: 1. A transient inward current carried by Na+ and Ca2+. This current is responsible for the upstroke of the action potentials. 2. A delayed outward current carried by K+. This current is voltage-sensitive and is responsible for the downstroke of the action potential during the early part of the burst. It becomes progressively inactivated during the burst. Its amplitude depends on the intracellular pH. 3. A rapidly developing outward current carried by K+ which is inactivated at potentials close to action potential threshold. This current tends to hold the membrane in the hyperpolarized state and is involved in spacing the action potentials. 4. A prolonged inward current which may not inactivate. It is probably carried by both Na+ and Ca2+. This current is responsible for the depolarizing phase of the burst but also contributes to the action potential. 5. A slowly developing outward current, carried by K+. This current appears as a result of a slow increase in intracellular ionized calcium and is responsible for the hyperpolarizing phase of the burst. Note that a transient increase in this current may also contribute to the falling phase of the action potential during the later stages of the burst. It is also sensitive to intracellular pH. One of the more significant features of this system of producing membrane potential oscillations is that the frequency of the bursts depends on the rate at which the intracellular ionized calcium returns to its resting level. This process depends on the metabolic state of the animal which can thereby exert a considerable influence on the electrical activity of burster neurones.


First metatarsophalangeal joint- MRI findings in asymptomatic volunteers.
To evaluate the spectrum and frequency of MR findings of the first metatarsophalangeal joint (MTPJ) in asymptomatic volunteers. MR imaging of 30 asymptomatic forefeet was performed with a dedicated extremity 1.5-Tesla system. Participants were between 20 and 49 years of age (mean ± SD: 35.5 ± 8.4 years). Two radiologists assessed cartilage, bone, capsuloligamentous structures, and tendons of first MTPJs on MR images. Cartilage defects were observed in 27 % (n = 8) of first MTPJs, most frequently located at the base of the proximal phalanx (23 %, n = 7), whereas cartilage defects of the metatarsal head (13 %, n = 4) and the metatarsosesamoid compartment were rare (0 %-3 %, n = 0-1). Bone marrow oedema-like signal changes were present in 37 % (n = 11) and subchondral cysts in 20 % (n = 6) of first MTPJs. Hyperintense areas on intermediate-weighted sequences (range: 30-43 %, n = 9-13) and on fluid-sensitive sequences with fat suppression (range: 33-60 %, n = 10-18) within the medial and lateral collateral ligament complex were common. Plantar recesses (77 %, n = 23) and distal dorsal recesses (87 %, n = 26) were frequently observed. Cartilage defects, bone marrow oedema-like signal changes, subchondral cysts, plantar recesses, and distal dorsal recesses were common findings on MRI of first MTPJs in asymptomatic volunteers. The collateral ligaments were often heterogeneous in structure and showed increased signal intensity. • Cartilage defects of asymptomatic first metatarsophalangeal joints were common on MRI. • The collateral ligaments were often heterogeneous in structure and showed increased signal intensity. • Areas of increased signal intensity within the flexor and extensor tendons were rare. • These observations need to be considered in MR examinations of symptomatic cases.


Clusterin is an ATP-independent chaperone with very broad substrate specificity that stabilizes stressed proteins in a folding-competent state.
We recently reported that the ubiquitous, secreted protein clusterin has chaperone activity in vitro [Humphreys et al. (1999) J. Biol. Chem. 274, 6875-6881]. In this study, we demonstrate that clusterin (i) inhibits stress-induced precipitation of a very broad range of structurally divergent protein substrates, (ii) binds irreversibly via an ATP-independent mechanism to stressed proteins to form solubilized high molecular weight complexes, (iii) lacks detectable ATPase activity, (iv) when acting alone, does not effect refolding of stressed proteins in vitro, and (v) stabilizes stressed proteins in a state competent for refolding by heat shock protein 70 (HSP70). Furthermore, we show that, at physiological levels, clusterin inhibits stress-induced precipitation of proteins in undiluted human serum. Clusterin represents the first identified secreted mammalian chaperone. However, reports from others suggest that, at least under stress conditions, clusterin may be retained within cells to exert a protective effect. Regardless of the topological site(s) of action, the demonstration that clusterin can stabilize stressed proteins in a refolding-competent state suggests that, during stresses, the action of clusterin may inhibit rapid and irreversible protein precipitation and produce a reservoir of inactive but stabilized molecules from which other refolding chaperones can subsequently salvage functional proteins.


Allelic reprogramming of 3D chromatin architecture during early mammalian development.
In mammals, chromatin organization undergoes drastic reprogramming after fertilization. However, the three-dimensional structure of chromatin and its reprogramming in preimplantation development remain poorly understood. Here, by developing a low-input Hi-C (genome-wide chromosome conformation capture) approach, we examined the reprogramming of chromatin organization during early development in mice. We found that oocytes in metaphase II show homogeneous chromatin folding that lacks detectable topologically associating domains (TADs) and chromatin compartments. Strikingly, chromatin shows greatly diminished higher-order structure after fertilization. Unexpectedly, the subsequent establishment of chromatin organization is a prolonged process that extends through preimplantation development, as characterized by slow consolidation of TADs and segregation of chromatin compartments. The two sets of parental chromosomes are spatially separated from each other and display distinct compartmentalization in zygotes. Such allele separation and allelic compartmentalization can be found as late as the 8-cell stage. Finally, we show that chromatin compaction in preimplantation embryos can partially proceed in the absence of zygotic transcription and is a multi-level hierarchical process. Taken together, our data suggest that chromatin may exist in a markedly relaxed state after fertilization, followed by progressive maturation of higher-order chromatin architecture during early development.


Irreversible regional cerebral ischemia: serial MR imaging and proton MR spectroscopy in a nonhuman primate model.
To delineate the changes in proton MR spectroscopy and imaging that occur with acute, irreversible ischemia of the basal ganglia of a baboon. The M1 segments of the middle cerebral arteries of six adult male baboons were occluded by endovascular means with microcatheters and N-butyl cyanoacrylate adhesive. Cerebral blood flow measurements were taken with positron emission tomography or radioactive microsphere techniques. Serial spatially localized proton MR spectroscopy of the basal ganglia and MR imaging of the brain were performed. The distribution of ischemic and infarcted tissue was demonstrated by histopathologic techniques or triphenyltetrazolium chloride staining. Radioactive microsphere or positron emission tomography measurements demonstrated no significant cerebral blood flow within the basal ganglia after occlusion of the middle cerebral artery. Proton MR spectroscopy of the basal ganglia demonstrated increasing cerebral lactate and decreasing N-acetyl aspartate within 30 minutes of middle cerebral artery occlusion. Changes in the MR imaging signal intensity of the basal ganglia were observed as early as 3.1 hours on T2-weighted, 3.3 hours on T1-weighted, and 6.1 hours on spin density-weighted images. The distribution of these changes correlated well with the histopathologic features of ischemia and infarction that were seen throughout the basal ganglia. Changes in MR imaging signal intensity corresponded to ischemia and infarction in our baboon model of acute irreversible ischemia of the basal ganglia. Increasing cerebral lactate and decreasing N-acetyl aspartate preceded changes in MR imaging signal intensity.


Antibiotics-First Versus Surgery for Appendicitis: A US Pilot Randomized Controlled Trial Allowing Outpatient Antibiotic Management.
Randomized trials suggest that nonoperative treatment of uncomplicated appendicitis with antibiotics-first is safe. No trial has evaluated outpatient treatment and no US randomized trial has been conducted, to our knowledge. This pilot study assessed feasibility of a multicenter US study comparing antibiotics-first, including outpatient management, with appendectomy. Patients aged 5 years or older with uncomplicated appendicitis at 1 US hospital were randomized to appendectomy or intravenous ertapenem greater than or equal to 48 hours and oral cefdinir and metronidazole. Stable antibiotics-first-treated participants older than 13 years could be discharged after greater than or equal to 6-hour emergency department (ED) observation with next-day follow-up. Outcomes included 1-month major complication rate (primary) and hospital duration, pain, disability, quality of life, and hospital charges, and antibiotics-first appendectomy rate. Of 48 eligible patients, 30 (62.5%) consented, of whom 16 (53.3%) were randomized to antibiotics-first and 14 (46.7%) to appendectomy. Median age was 33 years (range 9 to 73 years), median WBC count was 15,000/μL (range 6,200 to 23,100/μL), and median computed tomography appendiceal diameter was 10 mm (range 7 to 18 mm). Of 15 antibiotic-treated adults, 14 (93.3%) were discharged from the ED and all had symptom resolution. At 1 month, major complications occurred in 2 appendectomy participants (14.3%; 95% confidence interval [CI] 1.8% to 42.8%) and 1 antibiotics-first participant (6.3%; 95% CI 0.2% to 30.2%). Antibiotics-first participants had less total hospital time than appendectomy participants, 16.2 versus 42.1 hours, respectively. Antibiotics-first-treated participants had less pain and disability. During median 12-month follow-up, 2 of 15 antibiotics-first-treated participants (13.3%; 95% CI 3.7% to 37.9%) developed appendicitis and 1 was treated successfully with antibiotics; 1 had appendectomy. No more major complications occurred in either group. A multicenter US trial comparing antibiotics-first to appendectomy, including outpatient management, is feasible to evaluate efficacy and safety.


Mycorrhizal association as a primary control of the CO₂ fertilization effect.
Plants buffer increasing atmospheric carbon dioxide (CO2) concentrations through enhanced growth, but the question whether nitrogen availability constrains the magnitude of this ecosystem service remains unresolved. Synthesizing experiments from around the world, we show that CO2 fertilization is best explained by a simple interaction between nitrogen availability and mycorrhizal association. Plant species that associate with ectomycorrhizal fungi show a strong biomass increase (30 ± 3%, P < 0.001) in response to elevated CO2 regardless of nitrogen availability, whereas low nitrogen availability limits CO2 fertilization (0 ± 5%, P = 0.946) in plants that associate with arbuscular mycorrhizal fungi. The incorporation of mycorrhizae in global carbon cycle models is feasible, and crucial if we are to accurately project ecosystem responses and feedbacks to climate change.


Genetic Consideration of Schizotypal Traits: A Review.
Schizotypal traits are of interest and importance in their own right and also have theoretical and clinical associations with schizophrenia. These traits comprise attenuated psychotic symptoms, social withdrawal, reduced cognitive capacity, and affective dysregulation. The link between schizotypal traits and psychotic disorders has long since been debated. The status of knowledge at this point is such schizotypal traits are a risk for psychotic disorders, but in and of themselves only confer liability, with other risk factors needing to be present before a transition to psychosis occurs. Investigation of schizotypal traits also has the possibility to inform clinical and research pursuits concerning those who do not make a transition to psychotic disorders. A growing body of literature has investigated the genetic underpinnings of schizotypal traits. Here, we review association, family studies and describe genetic disorders where the expression of schizotypal traits has been investigated. We conducted a thorough review of the existing literature, with multiple search engines, references, and linked articles being searched for relevance to the current review. All articles and book chapters in English were sourced and reviewed for inclusion. Family studies demonstrate that schizotypal traits are elevated with increasing genetic proximity to schizophrenia and some chromosomal regions have been associated with schizotypy. Genes associated with schizophrenia have provided the initial start point for the investigation of candidate genes for schizotypal traits; neurobiological pathways of significance have guided selection of genes of interest. Given the chromosomal regions associated with schizophrenia, some genetic disorders have also considered the expression of schizotypal traits. Genetic disorders considered all comprise a profile of cognitive deficits and over representation of psychotic disorders compared to the general population. We conclude that genetic variations associated with schizotypal traits require further investigation, perhaps with targeted phenotypes narrowed to assist in refining the clinical end point of significance.


Quality of care after acute coronary syndromes in a prospective cohort with reasons for non-prescription of recommended medications.
Adherence to guidelines is associated with improved outcomes of patients with acute coronary syndrome (ACS). Clinical registries developed to assess quality of care at discharge often do not collect the reasons for non-prescription for proven efficacious preventive medication in Continental Europe. In a prospective cohort of patients hospitalized for an ACS, we aimed at measuring the rate of recommended treatment at discharge, using pre-specified quality indicators recommended in cardiologic guidelines and including systematic collection of reasons for non-prescription for preventive medications. In a prospective cohort with 1260 patients hospitalized for ACS, we measured the rate of recommended treatment at discharge in 4 academic centers in Switzerland. Performance measures for medication at discharge were pre-specified according to guidelines, systematically collected for all patients and included in a centralized database. Six hundred and eighty eight patients(54.6%) were discharged with a main diagnosis of STEMI, 491(39%) of NSTEMI and 81(6.4%) of unstable angina. Mean age was 64 years and 21.3% were women. 94.6% were prescribed angiotensin converting enzyme inhibitors/angiotensin II receptor blockers at discharge when only considering raw prescription rates, but increased to 99.5% when including reasons non-prescription. For statins, rates increased from 98% to 98.6% when including reasons for non-prescription and for beta-blockers, from 82% to 93%. For aspirin, rates further increased from 99.4% to 100% and from to 99.8% to 100% for P2Y12 inhibitors. We found a very high adherence to ACS guidelines for drug prescriptions at discharge when including reasons for non-prescription to drug therapy. For beta-blockers, prescription rates were suboptimal, even after taking into account reason for non-prescription. In an era of improving quality of care to achieve 100% prescription rates at discharge unless contra-indicated, pre-specification of reasons for non-prescription for cardiovascular preventive medication permits to identify remaining gaps in quality of care at discharge. ClinicalTrials.gov NCT01000701.


Epithelial origin of cutaneous anchoring fibrils.
Anchoring fibrils are essential structural elements of the dermoepidermal junction and are crucial to its functional integrity. They are composed largely of type VII collagen, but their cellular origin has not yet been confirmed. In this study, we demonstrate that the anchoring fibrils are primarily a product of epidermal keratinocytes. Human keratinocyte sheets were transplanted to a nondermal connective tissue graft bed in athymic mice. De novo anchoring fibril formation was studied ultrastructurally by immunogold techniques using an antiserum specific for human type VII procollagen. At 2 d after grafting, type VII procollagen/collagen was localized both intracellularly within basal keratinocytes and extracellularly beneath the discontinuous basal lamina. Within 6 d, a subconfluent basal lamina had developed, and newly formed anchoring fibrils and anchoring plaques subjacent to the xenografts were labeled. Throughout the observation period of the experiment, the maturity, population density, and architectural complexity of anchoring fibrils beneath the human epidermal graft continuously increased. Identical findings were obtained using xenografts cultivated from cloned human keratinocytes, eliminating the possibility of contributions to anchoring fibril regeneration from residual human fibroblasts. Immunolabeling was not observed at the mouse dermoepidermal junction at any time. These results demonstrate that the type VII collagen of human cutaneous anchoring fibrils and plaques is secreted by keratinocytes and can traverse the epidermal basal lamina and that the fibril formation can occur in the absence of cells of human dermal origin.


Social Psychological Face Perception: Why Appearance Matters.
We form first impressions from faces despite warnings not to do so. Moreover, there is considerable agreement in our impressions, which carry significant social outcomes. Appearance matters because some facial qualities are so useful in guiding adaptive behavior that even a trace of those qualities can create an impression. Specifically, the qualities revealed by facial cues that characterize low fitness, babies, emotion, and identity are overgeneralized to people whose facial appearance resembles the unfit (anomalous face overgeneralization), babies (babyface overgeneralization), a particular emotion (emotion face overgeneralization), or a particular identity (familiar face overgeneralization). We review studies that support the overgeneralization hypotheses and recommend research that incorporates additional tenets of the ecological theory from which these hypotheses are derived: the contribution of dynamic and multi-modal stimulus information to face perception; bidirectional relationships between behavior and face perception; perceptual learning mechanisms and social goals that sensitize perceivers to particular information in faces.


The NECAP PHear domain increases clathrin accessory protein binding potential.
AP-2 is a key regulator of the endocytic protein machinery driving clathrin-coated vesicle (CCV) formation. One critical function, mediated primarily by the AP-2 alpha-ear, is the recruitment of accessory proteins. NECAPs are alpha-ear-binding proteins that enrich on CCVs. Here, we have solved the structure of the conserved N-terminal region of NECAP 1, revealing a unique module in the pleckstrin homology (PH) domain superfamily, which we named the PHear domain. The PHear domain binds accessory proteins bearing FxDxF motifs, which were previously thought to bind exclusively to the AP-2 alpha-ear. Structural analysis of the PHear domain reveals the molecular surface for FxDxF motif binding, which was confirmed by site-directed mutagenesis. The reciprocal analysis of the FxDxF motif in amphiphysin I identified distinct binding requirements for binding to the alpha-ear and PHear domain. We show that NECAP knockdown compromises transferrin uptake and establish a functional role for NECAPs in clathrin-mediated endocytosis. Our data uncover a striking convergence of two evolutionarily and structurally distinct modules to recognize a common peptide motif and promote efficient endocytosis.


Estrogen Receptor β as a Therapeutic Target in Breast Cancer Stem Cells.
Breast cancer cells with tumor-initiating capabilities (BSCs) are considered to maintain tumor growth and govern metastasis. Hence, targeting BSCs will be crucial to achieve successful treatment of breast cancer. We characterized mammospheres derived from more than 40 cancer patients and two breast cancer cell lines for the expression of estrogen receptors (ERs) and stem cell markers. Mammosphere formation and proliferation assays were performed on cells from 19 cancer patients and five healthy individuals after incubation with ER-subtype selective ligands. Transcriptional analysis was performed to identify pathways activated in ERβ-stimulated mammospheres and verified using in vitro experiments. Xenograft models (n = 4 or 5 per group) were used to study the role of ERs during tumorigenesis. We identified an absence of ERα but upregulation of ERβ in BSCs associated with phenotypic stem cell markers and responsible for the proliferative role of estrogens. Knockdown of ERβ caused a reduction of mammosphere formation in cell lines and in patient-derived cancer cells (40.7%, 26.8%, and 39.1%, respectively). Gene set enrichment analysis identified glycolysis-related pathways (false discovery rate < 0.001) upregulated in ERβ-activated mammospheres. We observed that tamoxifen or fulvestrant alone was insufficient to block proliferation of patient-derived BSCs while this could be accomplished by a selective inhibitor of ERβ (PHTPP; 53.7% in luminal and 45.5% in triple-negative breast cancers). Furthermore, PHTPP reduced tumor initiation in two patient-derived xenografts (75.9% and 59.1% reduction in tumor volume, respectively) and potentiated tamoxifen-mediated inhibition of tumor growth in MCF7 xenografts. We identify ERβ as a mediator of estrogen action in BSCs and a novel target for endocrine therapy.


Transcriptional Activation of Arabidopsis Zygotes Is Required for Initial Cell Divisions.
Commonly referred to as the maternal-to-zygotic transition, the shift of developmental control from maternal-to-zygotic genomes is a key event during animal and plant embryogenesis. Together with the degradation of parental gene products, the increased transcriptional activities of the zygotic genome remodels the early embryonic transcriptome during this transition. Although evidence from multiple flowering plants suggests that zygotes become transcriptionally active soon after fertilization, the timing and developmental requirements of zygotic genome activation in Arabidopsis thaliana (Arabidopsis) remained a matter of debate until recently. In this report, we optimized an expansion microscopy technique for robust immunostaining of Arabidopsis ovules and seeds. This enabled the detection of marks indicative of active transcription in zygotes before the first cell division. Moreover, we employed a live-imaging culture system together with transcriptional inhibitors to demonstrate that such active transcription is physiologically required in zygotes and early embryos. Our results indicate that zygotic genome activation occurs soon after fertilization and is required for the initial zygotic divisions in Arabidopsis.


Regulator of G protein signaling 2 mediates cardiac compensation to pressure overload and antihypertrophic effects of PDE5 inhibition in mice.
The heart initially compensates for hypertension-mediated pressure overload by enhancing its contractile force and developing hypertrophy without dilation. Gq protein-coupled receptor pathways become activated and can depress function, leading to cardiac failure. Initial adaptation mechanisms to reduce cardiac damage during such stimulation remain largely unknown. Here we have shown that this initial adaptation requires regulator of G protein signaling 2 (RGS2). Mice lacking RGS2 had a normal basal cardiac phenotype, yet responded rapidly to pressure overload, with increased myocardial Gq signaling, marked cardiac hypertrophy and failure, and early mortality. Swimming exercise, which is not accompanied by Gq activation, induced a normal cardiac response, while Rgs2 deletion in Galphaq-overexpressing hearts exacerbated hypertrophy and dilation. In vascular smooth muscle, RGS2 is activated by cGMP-dependent protein kinase (PKG), suppressing Gq-stimulated vascular contraction. In normal mice, but not Rgs2-/- mice, PKG activation by the chronic inhibition of cGMP-selective phosphodiesterase 5 (PDE5) suppressed maladaptive cardiac hypertrophy, inhibiting Gq-coupled stimuli. Importantly, PKG was similarly activated by PDE5 inhibition in myocardium from both genotypes, but PKG plasma membrane translocation was more transient in Rgs2-/- myocytes than in controls and was unaffected by PDE5 inhibition. Thus, RGS2 is required for early myocardial compensation to pressure overload and mediates the initial antihypertrophic and cardioprotective effects of PDE5 inhibitors.


Understanding the Effects of Individual and State-Level Factors on American Public Response to COVID-19.
To examine multilevel predictors on American public response to COVID-19. Multilevel study. A national survey was conducted by Qualtrics from August 24 to September 11, 2020. The state-level variables were constructed on data from multiple sources. 2,440 respondents 18 years and older from all 50 states and D.C. The outcome variable is the public response to COVID-19 measured by threat perception, behavioral adjustment, and policy support. The predictors include individual-level sociodemographic factors and state-level indicators about public health conditions, political context, and economic recovery. Multilevel structural equation modeling is used for statistical estimation. People from states with more COVID-19 cases (β = 0.020, p < 0.1), mandatory face mask policies (β = 0.069, p < 0.05), and liberal governments (β = 0.002, p < 0.05) are more likely to respond while people from states whose economies have recovered closer to the pre-pandemic level are less likely to do so (β = -0.005, p < 0.05). Regarding individual-level predictors, older people (β = 0.005, p < 0.001) and people with better education (β = 0.029, p < 0.01), leaning toward the Democrat Party (β = 0.066, p < 0.001) and liberal political ideology (β = 0.094, p < 0.001), and have stronger generalized trust (β = 0.033, p < 0.001) are more likely to respond than their counterparts. Differences in the public response to the pandemic stem from variations in individual characteristics and contextual factors of states where people live. These findings contribute to the rapidly growing literature and have implications for public health policies.


Somatic mosaicism for copy number variation in differentiated human tissues.
Two major types of genetic variation are known: single nucleotide polymorphisms (SNPs), and a more recently discovered structural variation, involving changes in copy number (CNVs) of kilobase- to megabase-sized chromosomal segments. It is unknown whether CNVs arise in somatic cells, but it is, however, generally assumed that normal cells are genetically identical. We tested 34 tissue samples from three subjects and, having analyzed for each tissue < or =10(-6) of all cells expected in an adult human, we observed at least six CNVs, affecting a single organ or one or more tissues of the same subject. The CNVs ranged from 82 to 176 kb, often encompassing known genes, potentially affecting gene function. Our results indicate that humans are commonly affected by somatic mosaicism for stochastic CNVs, which occur in a substantial fraction of cells. The majority of described CNVs were previously shown to be polymorphic between unrelated subjects, suggesting that some CNVs previously reported as germline might represent somatic events, since in most studies of this kind, only one tissue is typically examined and analysis of parents for the studied subjects is not routinely performed. A considerable number of human phenotypes are a consequence of a somatic process. Thus, our conclusions will be important for the delineation of genetic factors behind these phenotypes. Consequently, biobanks should consider sampling multiple tissues to better address mosaicism in the studies of somatic disorders.


In vivo Evidence for Therapeutic Properties of Cannabidiol (CBD) for Alzheimer's Disease.
Alzheimer's disease (AD) is a debilitating neurodegenerative disease that is affecting an increasing number of people. It is characterized by the accumulation of amyloid-β and tau hyperphosphorylation as well as neuroinflammation and oxidative stress. Current AD treatments do not stop or reverse the disease progression, highlighting the need for new, more effective therapeutics. Cannabidiol (CBD) is a non-psychoactive phytocannabinoid that has demonstrated neuroprotective, anti-inflammatory and antioxidant properties in vitro. Thus, it is investigated as a potential multifunctional treatment option for AD. Here, we summarize the current status quo of in vivo effects of CBD in established pharmacological and transgenic animal models for AD. The studies demonstrate the ability of CBD to reduce reactive gliosis and the neuroinflammatory response as well as to promote neurogenesis. Importantly, CBD also reverses and prevents the development of cognitive deficits in AD rodent models. Interestingly, combination therapies of CBD and Δ9-tetrahydrocannabinol (THC), the main active ingredient of cannabis sativa, show that CBD can antagonize the psychoactive effects associated with THC and possibly mediate greater therapeutic benefits than either phytocannabinoid alone. The studies provide "proof of principle" that CBD and possibly CBD-THC combinations are valid candidates for novel AD therapies. Further investigations should address the long-term potential of CBD and evaluate mechanisms involved in the therapeutic effects described.


Direct estimation of death attributable to smoking in Switzerland based on record linkage of routine and observational data.
In Switzerland, estimations of smoking-attributable deaths were based on age- and sex-adjusted hazard ratios (HRs) from foreign cohorts, precluding consideration of country-specific properties and adjustment for confounding. In order to overcome this, we analyzed recently available individual data from Switzerland. We included 17,861 individuals aged ≥16 years who participated between 1977-1993 in health studies and were anonymously linked with the Swiss National Cohort. Adjusted Cox regression was used to calculate mortality HRs. Smoking status at baseline was categorized into never-smokers, former smokers, and current light or heavy smokers (<20 or ≥20 cigarettes/day). As covariates, we selected education, marital status, lifestyle, alcohol consumption, and body mass index. We differentiated between cardiovascular disease (CVD), cancer, and noncancer-non-CVD deaths. Smoking-attributable deaths were estimated with a HR-based approach and with age-specific prevalence rates and mortality estimates from 2007. Smoking men and women not only had an increased risk for all-cause (HR and 95% confidence interval vs. never-smokers: 1.71 [1.53-1.90]; 1.54 [1.36-1.75]), CVD (1.72 [1.43-2.06]; 1.50 [1.19-1.90]) and cancer (1.87 [1.56-2.25]; 1.58 [1.30-1.93]), but also for noncancer-non-CVD death (1.57 [1.29-1.89]; 1.58 [1.30-1.93]). Former smoking men had an increased risk for all-cause (1.16 [1.03-1.31]) and cancer death (1.35 [1.10-1.65]). Multivariate adjustment only slightly modified the association between smoking and mortality. Overall, 7,153 deaths per year could be attributed to smoking. Smoking is an important avoidable health burden in Switzerland, and its consequences may persist for decades after quitting. This stresses the need for putting more efforts in strategies aimed at preventing the onset of smoking.


Mechanisms of Histone Deacetylase Inhibitor-Regulated Gene Expression in Cancer Cells.
Class I and II histone deacetylase inhibitors (HDACis) are approved for the treatment of cutaneous T-cell lymphoma and are undergoing clinical trials as single agents, and in combination, for other hematological and solid tumors. Understanding their mechanisms of action is essential for their more effective clinical use, and broadening their clinical potential. HDACi induce extensive transcriptional changes in tumor cells by activating and repressing similar numbers of genes. These transcriptional changes mediate, at least in part, HDACi-mediated growth inhibition, apoptosis, and differentiation. Here, we highlight two fundamental mechanisms by which HDACi regulate gene expression—histone and transcription factor acetylation. We also review the transcriptional responses invoked by HDACi, and compare these effects within and across tumor types. The mechanistic basis for how HDACi activate, and in particular repress gene expression, is not well understood. In addition, whether subsets of genes are reproducibly regulated by these agents both within and across tumor types has not been systematically addressed. A detailed understanding of the transcriptional changes elicited by HDACi in various tumor types, and the mechanistic basis for these effects, may provide insights into the specificity of these drugs for transformed cells and specific tumor types. Understanding the mechanisms by which HDACi regulate gene expression and an appreciation of their transcriptional targets could facilitate the ongoing clinical development of these emerging therapeutics. In particular, this knowledge could inform the design of rational drug combinations involving HDACi, and facilitate the identification of mechanism-based biomarkers of response.


Cytogenetic heterogeneity of translocations associated with Duchenne muscular dystrophy.
Lymphoblastoid cell lines have been established from nine female patients with Duchenne muscular dystrophy who had previously been reported to have chromosome translocations with breakpoints in the Xp21 region. A detailed cytogenetic comparison of prometaphase chromosomes in these cell lines revealed that six of the translocations had X chromosome breakpoints in the sub-band Xp212 and that one further breakpoint could be assigned to either Xp212 or Xp213. These findings confirm and extend previous observations and provide strong evidence for Xp212 as the site of the Duchenne and Becker loci. For the remaining two translocations the simplest explanation for the observed banding pattern is that the X chromosome breakpoint lies a few thousand kilobases away, in the sub-band Xp211. Other explanations which assume breaks in Xp212 combined with complex local chromosome rearrangements are also presented. It is also possible that the altered banding pattern in these two cases is due to the influence of local sequences on the staining or uncoiling properties of the chromatin.


Circulating free mitochondrial DNA concentration and its association with erlotinib treatment in patients with adenocarcinoma of the lung.
Changes in circulating free DNA concentrations have been correlated with chemotherapeutic effects in solid tumors. The present study was designed to determine and compare the changes in circulating free mitochondrial DNA (mtDNA) concentrations prior to and following erlotinib treatment, as well as the potential prognostic value of plasma mtDNA. Patients with adenocarcinoma of the lung who were to receive erlotinib treatment were enrolled in the present study once informed consent had been obtained. Patient plasma samples were collected immediately prior to starting erlotinib treatment, on days 15 and 29 following the initiation of erlotinib treatment and also when the patient's disease had progressed. The most common erlotinib treatment response was a partial response (PR), achieved in 26 (49.1%) of the 53 enrolled patients, followed by stable disease (SD) in 13 patients (24.5%) and progressive disease (PD) in 14 patients (26.4%). Plasma mtDNA concentrations were significantly decreased on day 15 compared with day 0 in the patients with PD (P=0.028) or in those patients without a response to erlotinib treatment (SD and PD; P=0.007). Plasma mtDNA concentrations were similar or elevated on day 15 compared with day 0 in the patients with a PR (P=0.808). The concentration of plasma mtDNA did not correlate with progression-free survival (PFS). Tumor epidermal growth factor receptor (EGFR) mutation status (activating mutations in 16 patients and wild-type in 14 patients) did not correlate with the concentration of plasma mtDNA (P=0.951). Plasma mtDNA levels did not correlate with the PFS of the patients when they received erlotinib treatment. The plasma mtDNA levels were decreased on day 15 in those patients who had disease progression following erlotinib treatment. These results demonstrate that plasma mtDNA is of weak clinical utility as a screening, diagnostic or prognostic tool in lung cancer patients.


Taking advantage of luminescent lanthanide ions.
Lanthanide ions possess fascinating optical properties and their discovery, first industrial uses and present high technological applications are largely governed by their interaction with light. Lighting devices (economical luminescent lamps, light emitting diodes), television and computer displays, optical fibres, optical amplifiers, lasers, as well as responsive luminescent stains for biomedical analysis, medical diagnosis, and cell imaging rely heavily on lanthanide ions. This critical review has been tailored for a broad audience of chemists, biochemists and materials scientists; the basics of lanthanide photophysics are highlighted together with the synthetic strategies used to insert these ions into mono- and polymetallic molecular edifices. Recent advances in NIR-emitting materials, including liquid crystals, and in the control of luminescent properties in polymetallic assemblies are also presented. (210 references.).


Highly potent and selective phenylmorphan-based inverse agonists of the opioid delta receptor.
We recently reported the discovery of (+)-5-(3-hydroxyphenyl)-4-methyl-2-(3-phenylpropyl)-2-azabicyclo[3.3.1]non-7-yl-(1-phenyl-1-cyclopentane)carboxamide [(+)-KF4, (+)-5] as a novel chemotype possessing potent antagonist activity at the delta opioid receptor. Additional SAR studies involving changes to both the 2-amino and 7-amido N-substituents using this same (+)-morphan scaffold have revealed compounds with improved potency and selectivity for the delta opioid receptor. The highly potent and selective 2,2-dimethylphenylacetamide analogue (+)-N-[(1S,4R,5R,7S)-5-(3-hydroxyphenyl)-4-methyl-2-(3-phenylpropyl)-2-azabicyclo[3.3.1]non-7-yl]-2-methyl-2-phenylpropanamide (13d, delmorphan-A) showed picomolar inhibitory potency (Ke = 0.1 nM) in the [35S]GTPgammaS functional assay with delta opioid receptor selectivity ratios of 103- and 132-fold versus the mu and kappa opioid receptors, respectively. The compounds showed no agonist activity at any of the three opioid receptors; however, measurements of delta inverse agonist activity within this series illustrated a broad range of negative efficacy and IC50 values 650-fold more potent than the prototypical delta opioid receptor inverse agonist ICI 174,864 (22).


Illness, body and relationship: Giovanni Jervis and the field of clinical psychology.
Unlike most of his contemporaries, Jervis felt it was necessary to give new answers to last century's clinical psychology crisis. He managed to show the relevance that psychological knowledge coming from other sectors (such as philosophy, evolutionary biology, cybernetics, ethology and, nowadays, neuroscience) could have for clinical psychology. He realized that the clinical world was at risk of loosing its cultural models of reference and threatened to break up with the scientific tradition with which it had not been able to establish a profitable dialogue. Jervis' views on mental functioning allowed him to delimit the field of action of clinical psychology. The focus on pathology and existential distress suggested that correct clinical evaluation and diagnosis are at the core of the activity of clinical psychologists. The tension between relational expectations, self-deceptive aspects of identity and the bodily sources of mental life show the dynamic field on which clinical psychology can deploy its transformative action.


Oxidative modification of hepatic mitochondria protein thiols: effect of chronic alcohol consumption.
Redox modification of mitochondrial proteins is thought to play a key role in regulating cellular function, although direct evidence to support this hypothesis is limited. Using an in vivo model of mitochondrial redox stress, ethanol hepatotoxicity, the modification of mitochondrial protein thiols was examined using a proteomics approach. Specific labeling of reduced thiols in the mitochondrion from the livers of control and ethanol-fed rats was achieved by using the thiol reactive compound (4-iodobutyl)triphenylphosphonium (IBTP). This molecule selectively accumulates in the organelle and can be used to identify thiol-containing proteins. Mitochondrial proteins that have been modified are identified by decreased labeling with IBTP using two-dimensional SDS-PAGE followed by immunoblotting with an antibody directed against the triphenylphosphonium moiety of the IBTP molecule. Analyses of these data showed a significant decrease in IBTP labeling of thiols present in specific mitochondria matrix proteins from ethanol-fed rats compared with their corresponding controls. These proteins were identified as the low-K(m) aldehyde dehydrogenase and glucose-regulated protein 78. The decrease in IBTP labeling in aldehyde dehydrogenase was accompanied by a decrease in specific activity of the enzyme. These data demonstrate that mitochondrial protein thiol modification is associated with chronic alcohol intake and might contribute to the pathophysiology associated with hepatic injury. Taken together, we have developed a protocol to chemically tag and select thiol-modified proteins that will greatly enhance efforts to establish posttranslational redox modification of mitochondrial protein in in vivo models of oxidative or nitrosative stress.


Insulin-like growth factor type 1 upregulates uncoupling protein 3.
In this study the expression of uncoupling protein 3 (UCP3) and its regulation by insulin-like growth factor 1 (IGF-I) and insulin in human neuroblastoma SH-SY5Y cells were characterized. Reverse transcriptase-PCR, Western blot, and immunofluorescence analysis showed that SH-SY5Y cells express UCP3 natively. IGF-I induced a time- and concentration-dependent induction of UCP3 protein reaching a twofold expression after 72 h with 10 nM IGF-I. Extremely high insulin concentrations (860 nM) and 10 nM trIGF-I, a truncated form of IGF-I with the same affinity for the IGF-I receptor as the full-length IGF-I, but with lower activity on the insulin receptor, also upregulated UCP3. We conclude that SH-SY5Y cells express UCP3 natively and that the expression is regulated by IGF-I via the IGF-I receptor.


Health surveillance of preschool children.
Discussions with every general practice, health visitor, and clinical medical officer in Northumberland Health Authority led to agreement about the content of preschool health surveillance, the ages at which it should be done, and referral pathways after a failed screening test. Each primary health care team now undertakes to do a basic minimum set of screening tests, and each team decides who in the team will do each test. The screening system agreed on should enable time to become available for the equally important aspects of surveillance--namely, developmental guidance, health education, and assessment and follow up of problems. The discussions also led to agreement about how the health authority should evaluate the effect of the surveillance programme on the health of children.


The circadian clock functions as a potent regulator of allergic reaction.
Symptoms and laboratory parameters of allergic diseases exhibit prominent ~24-h variations. For instance, in most allergic rhinitis patients, symptoms worsen overnight or early in the morning. Accordingly, there are benefits to nighttime dosing of anti-allergy medications in such patients. Although the circadian pathophysiology of allergic diseases is well documented, the biological basis of this phenomenon remains poorly understood. Recent studies have begun to reveal that the internal timekeeping system termed the circadian clock plays a key role in temporal regulation of allergic reaction, and may therefore underlie the circadian pathophysiology of allergic diseases. Here, we review new knowledge that highlights the emerging role of the circadian clock as a potent regulator of allergic reactions. Given the strong influence of circadian rhythms on allergic diseases, we believe that research on how the time of day impacts allergic reaction which we may call 'chronoallergology' will provide new insight into previously unknown aspects of the biology of allergies. Such knowledge should facilitate novel strategies for prevention and treatment of these diseases.


Key formulation variables in tableting of coated pellets.
Multiple unit controlled release dosage forms offer various advantages over their single unit counterparts. Most of these advantages are associated with the uniform distribution of multiparticulates throughout the gastrointestinal tract. Though coated pellets can be filled into hard gelatin capsules, tablet formulation is the preferred one because of various advantages associated with it. However, compression of coated pellets is a challenging task necessitating the optimization of various formulation and process variables. The key formulation variables include composition, porosity, size, shape and density of the pellets; type and amount of polymer coating; nature, size and amount of tableting excipients. The pellet core should be strong with some degree of plasticity. It should be highly porous, small, with an irregular shape. The critical density to achieve prolonged release was reported to lie between 2.4 and 2.8 g/cm(3). Acrylic polymer films are more flexible and more suitable for the coating of pellets to be compressed into tablets. Thicker coatings offer better resistance to frictional forces. Solvent based coatings are more flexible and have a higher degree of mechanical stability than the aqueous based ones. The tableting excipients should have cushioning property. They should not be significantly different in size and density from those of the pellet cores in order to avoid segregation. Addition of 30%-60% of tableting excipients is necessary to avoid any damage to the polymer coat and to retain its functional property.


Inactivation of the ptsI gene encoding enzyme I of the sugar phosphotransferase system of Streptococcus salivarius: effects on growth and urease expression.
The urease genes of Streptococcus salivarius 57.1 are tightly repressed in cells growing at neutral pH. When cells are cultivated at acidic pH values, the urease genes become derepressed and transcription is enhanced when cells are growing under carbohydrate-excess conditions. Previously, the authors proposed that the bacterial sugar:phosphotransferase system (PTS) modulated the DNA-binding activity by phosphorylation of the urease repressor when carbohydrate was limiting. The purpose of this study was to assess whether enzyme I (EI) of the PTS could be involved in modulating urease expression in response to carbohydrate availability. An EI-deficient strain (ptsI18-3) of S. salivarius 57.1 was constructed by insertional inactivation of the ptsI gene. The mutant had no measurable PTS activity and lacked EI, as assessed by Western analysis. The mutant grew as well as the wild-type strain on the non-PTS sugar lactose, and grew better than the parent when another non-PTS sugar, galactose, was the sole carbohydrate. The mutant was able to grow with glucose as the sole carbohydrate, but displayed a 24 h lag time and had a generation time some threefold longer than strain 57.1. The mean OD600 attained after 48 h by ptsI18-3 supplied with fructose was 0.16, with no additional growth observed even after 3 d. Urease expression in the wild-type and mutant strains was assessed in continuous chemostat culture. Repression of urease at neutral pH was seen in both strains under all conditions tested. Growth of wild-type cells on limiting concentrations of lactose resulted in very low levels of urease expression compared with growth on PTS sugars. In contrast, under similar conditions, urease expression in ptsI18-3 was restored to levels seen in the parent growing on PTS sugars. Growth under conditions of lactose excess resulted in further derepression of urease, but ptsI18-3 expressed about threefold higher urease activity than 57.1. The results support a role for EI in urease regulation, but also indicate that additional factors may be important in regulating urease gene expression.


Diabetes and Its Effect on Abdominal Aortic Aneurysm Growth Rate in Hispanic Patients.
The growth rate of abdominal aortic aneurysms (AAA) can vary depending on age, baseline diameter, blood pressure, race, and history of smoking. Paradoxically, previous studies show evidence of a protective effect of diabetes on the rate of AAA expansion despite its well-established role in the morbidity and mortality of cardiovascular disease. This study aims to investigate the impact diabetes plays on AAA growth within a Hispanic population. Data were collected from patients who were predominantly Mexican-American at a single hospital site. Baseline and follow-up measures for AAA diameter were obtained from serial imaging studies. Demographics, medical history, the presence of type 2 diabetes, and medication use were extracted from hospital records. Linear mixed-effects growth models were used to calculate the overall AAA growth rate and to assess the difference in AAA growth rate between demographics, comorbidities, and medication use. The study comprised 201 patients (70.4% male) with a mean baseline age of 79.1 years, of whom 43.2% were diabetic. The average monthly AAA growth rate across all study participants was 0.15 mm (SE = 0.02 mm). Independently, the average AAA expansion rate for the diabetic and nondiabetic groups was 0.07 mm (SE = 0.04 mm) and 0.21 mm (SE = 0.03 mm) per month, respectively. This demonstrates a 65% lower linear AAA expansion rate per month in patients with diabetes. This study confirms a difference of AAA physiology between diabetics and nondiabetics in the Hispanic community. The observed significant difference in AAA growth rate may be a combination of factors associated with race/ethnicity, prevalence of diabetes mellitus, and low compliance with diabetic control exhibited in the Mexican-American population.


Effectiveness of using a behavioural intervention to improve dietary fibre intakes in children with constipation.
Functional constipation is common in childhood; diets in affluent industrialised countries may be a significant contributory factor as a result of inadequate fibre content. Only a few intervention studies have been reported where childhood constipation has been treated by increasing dietary fibre. The present study aimed to demonstrate that dietary fibre intakes of children with constipation can be increased using a specifically-designed behaviour modification technique with a self-monitoring and reward system. Forty-three children, aged 2-14years, with functional constipation (defined as less than three bowel movements per week with hard stools and difficulty or delay in defecation) were randomised to one of two treatment groups: Control (n=20), receiving general advice on increasing dietary fibre intake, or Intervention (n=23), using the intervention tool. Fibre intake, laxative use and stool frequency were assessed at baseline, and at 3, 6 and 12months, using parent-filled diaries. A behavioural intervention method significantly increases the fibre intakes of children with constipation at 3 months compared to standard dietary treatment (P=0.005), remaining so after adjusting for baseline fibre intake (P=0.007). Follow-up at 6 and 12months showed no further increase. No significant benefit in terms of a reduction in laxative use or increased stool frequency associated with additional fibre intake was demonstrated. The findings confirm the difficulties encountered in beginning and maintaining high-fibre diets in children.


Automatic analysis of speckle photographs with extended range and improved accuracy.
A new algorithm for automatic analysis of Young's fringes in speckle photography, applicable to fringe numbers >0.5, is developed. In the algorithm we first divide the diffraction pattern by the corresponding diffraction halo, then integrate the result over bands in various directions for detecting the fringe orientation, and use curve fitting for determining the fringe spacing. The algorithm has enabled fully automatic analysis of double-exposure specklegrams for a wide range of speckle displacement with improved accuracy. Contours of displacement and strain fields have also been plotted using the result of the analysis.


The Relationship between 25 (OH) D Levels (Vitamin D) and Bone Mineral Density (BMD) in a Saudi Population in a Community-Based Setting.
Vitamin D deficiency has been linked to an increased risk of osteoporosis. Vitamin D deficiency has reached high levels in the Saudi population, but there is conflicting evidence both in the Saudi population, and worldwide, regarding the existence of a correlation between these low vitamin D levels and reduced BMD (bone mineral density), or osteoporosis. The objective of this study was primarily to determine whether there was a correlation between vitamin D deficiency and osteoporosis in the Saudi population. We aimed to investigate whether the high levels of vitamin D deficiency and insufficiency would translate to higher prevalence of osteoporosis, and whether there is a correlation between vitamin D levels and bone mineral density. This was a community based cross sectional study conducted in the Family Medicine Clinics at King Faisal Specialist Hospital and Research Centre in Riyadh, Saudi Arabia. Electronic records of 1723 patients were reviewed. Laboratory and radiology results were collected, including vitamin D levels, calcium levels, and bone mineral density scan results. Among the whole population, 61.5% had moderate to severe vitamin D deficiency with levels less than 50nmol/L. 9.1% of the population had osteoporosis, and 38.6% had osteopenia. Among the whole population, there was no significant correlation between spine or total femoral BMD and serum 25(OH) D. Vitamin D deficiency is prevalent in the Saudi population. However, no correlation has been found between vitamin D deficiency and reduced bone mineral density in any age group, in males or females, Saudis or Non-Saudis, in our population in Riyadh, Saudi Arabia.


Social skills groups for people aged 6 to 21 with autism spectrum disorders (ASD).
Since autism was first described, major difficulties in social interaction have been a defining feature of individuals with autism spectrum disorders (ASD). Social skills groups are a common intervention for individuals with ASD. Although a frequently recommended practice, the few studies that have addressed the efficacy of social skills groups have shown mixed results. To determine the effectiveness of social skills groups for improving social competence, social communication, and quality of life for people with ASD who are six to 21 years of age. We searched the following databases in December 2011: CENTRAL (2011 Issue 4), MEDLINE (1948 to November Week 3, 2011), EMBASE (1980 to Week 50, 2011), PsycINFO (1887 to December Week 2, 2011), CINAHL (1937 to current), ERIC (1966 to current), Sociological Abstracts (1952 to current), OCLC WorldCat (12 December 2011), Social Science Citation Index (1970 to 16 December 2011), and the metaRegister of Controlled Trials (20 December 2011). We also searched the reference lists of published papers. Randomized control trials (RCTs) comparing treatment (social skills groups) with a control group who were not receiving the treatment for participants aged six to 21 years with ASD. The control group could be no intervention, wait list, or treatment as usual. Outcomes sought were standardized measures of social competence, social communication, quality of life, emotion recognition, and any other specific behaviors. Two review authors independently selected and appraised studies for inclusion and assessed the risk of bias in each included study. All outcome data were continuous and standardized mean difference effect sizes (ES) with small sample correction were calculated. We conducted random-effects meta-analysis where possible. We included five RCTs evaluating the effects of social skills groups in 196 participants with ASD aged 6 to 21 years old. The results show there is some evidence that social skills groups improve overall social competence (ES = 0.47, 95% confidence interval (CI) 0.16 to 0.78, P = 0.003) and friendship quality (ES = 0.41, 95% CI 0.02 to 0.81, P = 0.04) for this population. No differences were found between treatment and control groups in relation to emotional recognition (ES = 0.34, 95% CI -0.20 to 0.88, P = 0.21) assessed in two studies or social communication as related to the understanding of idioms (ES = 0.05, 95% CI -0.63 to 0.72, P = 0.89), which was assessed in only one study. Two additional quality of life outcomes were evaluated, with results of single studies suggesting decreases in loneliness (ES = -0.66, 95% CI -1.15 to -0.17) but no effect on child or parental depression. No adverse events were reported.Given the nature of the intervention and the selected outcome measures, the risk of performance and detection bias are high. There is limited generalizability from the studies as they were all conducted in the US; they focused mainly on children aged 7 to 12, and the participants were all of average or above average intelligence. There is some evidence that social skills groups can improve social competence for some children and adolescents with ASD. More research is needed to draw more robust conclusions, especially with respect to improvements in quality of life.


A Logic Model of Neuronal-Glial Interaction Suggests Altered Homeostatic Regulation in the Perpetuation of Neuroinflammation.
Aberrant inflammatory signaling between neuronal and glial cells can develop into a persistent sickness behavior-related disorders, negatively impacting learning, memory, and neurogenesis. While there is an abundance of literature describing these interactions, there still lacks a comprehensive mathematical model describing the complex feed-forward and feedback mechanisms of neural-glial interaction. Here we compile molecular and cellular signaling information from various studies and reviews in the literature to create a logically-consistent, theoretical model of neural-glial interaction in the brain to explore the role of neuron-glia homeostatic regulation in the perpetuation of neuroinflammation. Logic rules are applied to this connectivity diagram to predict the system's homeostatic behavior. We validate our model predicted homeostatic profiles against RNAseq gene expression profiles in a mouse model of stress primed neuroinflammation. A meta-analysis was used to calculate the significance of similarity between the inflammatory profiles of mice exposed to diisopropyl fluorophostphate (DFP) [with and without prior priming by the glucocorticoid stress hormone corticosterone (CORT)], with the equilibrium states predicted by the model, and to provide estimates of the degree of the neuroinflammatory response. Beyond normal homeostatic regulation, our model predicts an alternate self-perpetuating condition consistent with chronic neuroinflammation. RNAseq gene expression profiles from the cortex of mice exposed to DFP and CORT+DFP align with this predicted state of neuroinflammation, whereas the alignment to CORT alone was negligible. Simulations of putative treatment strategies post-exposure were shown to be theoretically capable of returning the system to a state of typically healthy regulation with broad-acting anti-inflammatory agents showing the highest probability of success. The results support a role for the brain's own homeostatic drive in perpetuating the chronic neuroinflammation associated with exposure to the organophosphate DFP, with and without CORT priming. The deviation of illness profiles from exact model predictions suggests the presence of additional factors or of lasting changes to the brain's regulatory circuitry specific to each exposure.


COMBATdb: a database for the COVID-19 Multi-Omics Blood ATlas.
Advances in our understanding of the nature of the immune response to SARS-CoV-2 infection, and how this varies within and between individuals, is important in efforts to develop targeted therapies and precision medicine approaches. Here we present a database for the COvid-19 Multi-omics Blood ATlas (COMBAT) project, COMBATdb (https://db.combat.ox.ac.uk). This enables exploration of multi-modal datasets arising from profiling of patients with different severities of illness admitted to hospital in the first phase of the pandemic in the UK prior to vaccination, compared with community cases, healthy controls, and patients with all-cause sepsis and influenza. These data include whole blood transcriptomics, plasma proteomics, epigenomics, single-cell multi-omics, immune repertoire sequencing, flow and mass cytometry, and cohort metadata. COMBATdb provides access to the processed data in a well-defined framework of samples, cell types and genes/proteins that allows exploration across the assayed modalities, with functionality including browse, search, download, calculation and visualisation via shiny apps. This advances the ability of users to leverage COMBAT datasets to understand the pathogenesis of COVID-19, and the nature of specific and shared features with other infectious diseases.


Spillover to triadic and dyadic systems in families with young children.
Research has evidenced support for the spillover model, which asserts that parents' marital functioning influences their parenting and coparenting behavior in dyadic (mother-child and father-child) and triadic (mother-father-child) family contexts. However, few studies have simultaneously investigated the spillover model in both parenting and coparenting systems, preventing examination of whether spillover impacts both systems equally or differentially. Further, little research has examined whether quality of the marital system influences children's behavior toward their parents, as well as their parents' behavior, in dyadic interactions. We examined the spillover model using observational measures of parent and child behavior in parent-child dyadic interactions as well as coparenting in triadic interactions. We also explored parent and child gender differences in spillover effects. Participants were families with children aged 3 to 6 years (n = 149). Findings indicated that spillover occurs to multiple family systems, but the effects varied according to whose behavior (mother, father, child) was explored. In families of boys and girls, the marital system influenced warmth in triadic interactions, as well as fathers' responsiveness and children's responsiveness to mothers in dyadic interactions. Spillover effects were largely equivalent for girls and boys, but spillover to coparenting hostility in triadic interactions was limited to families raising girls. Parent gender also moderated associations between marital functioning and parent-child interactions: Spillover was significantly stronger for fathers' responsiveness (vs. mothers' responsiveness) and child responsiveness to mothers (vs. child responsiveness to fathers).


Improving clinical trial outcomes in amyotrophic lateral sclerosis.
Individuals who are diagnosed with amyotrophic lateral sclerosis (ALS) today face the same historically intransigent problem that has existed since the initial description of the disease in the 1860s - a lack of effective therapies. In part, the development of new treatments has been hampered by an imperfect understanding of the biological processes that trigger ALS and promote disease progression. Advances in our understanding of these biological processes, including the causative genetic mutations, and of the influence of environmental factors have deepened our appreciation of disease pathophysiology. The consequent identification of pathogenic targets means that the introduction of effective therapies is becoming a realistic prospect. Progress in precision medicine, including genetically targeted therapies, will undoubtedly change the natural history of ALS. The evolution of clinical trial designs combined with improved methods for patient stratification will facilitate the translation of novel therapies into the clinic. In addition, the refinement of emerging biomarkers of therapeutic benefits is critical to the streamlining of care for individuals. In this Review, we synthesize these developments in ALS and discuss the further developments and refinements needed to accelerate the introduction of effective therapeutic approaches.


DET1 and COP1 Modulate the Coordination of Growth and Immunity in Response to Key Seasonal Signals in Arabidopsis.
Plant growth and development and outcomes of plant-microbe interactions are defined by coordinated responses to seasonal signals. The mechanisms that control the coordinated regulation of growth and immunity are not well understood. Here, we show that a common signaling module integrates environmental signals, such as photoperiod and temperature, to regulate the growth-defense balance. Key light-signaling components De-Etiolated 1 (DET1) and Constitutive Photomorphogenic 1 (COP1) negatively regulate immunity and are essential for immune modulation by photoperiod and temperature. Our results show that this is regulated by the transcription factor Phytochrome Interacting Factor 4 (PIF4), suggesting that the DET1/COP1-PIF4 module acts as a central hub for the control of growth and immunity in response to seasonal signals. These findings provide a regulatory framework for environmental signal integration.


Homogeneity of fascicle architecture following repeated contractions in the human gastrocnemius medialis.
This investigation sought to determine the effects of fatigue on fascicle architecture across the length of the human gastrocnemius medialis (GM). With institutional ethical approval, fifteen healthy males performed repeated isometric plantar flexion maximal voluntary contractions (MVC) until peak force fell 30% below baseline. Brightness-mode ultrasound was used to determine fascicle length and pennation angle at rest and during MVC prior to and following the fatiguing contractions. The results show a significant increase in fascicle length during MVC in the distal (2.8 mm, 8.1%) middle, (4.9 mm, 14.1%), and proximal (5.2 mm, 14.7%) regions post-fatigue compared to pre-fatigue (p < 0.05). There was also a significant decrease of pennation angle during MVC in the distal (3.3°, 8.8%), middle (3.9°, 9.4%), and proximal (2.9°, 6.9%) regions post-fatigue compared to pre-fatigue (p < 0.05). These changes, however, were not region specific. These are the first results to show that fascicle shortening within the GM remains homogeneous following fatigue, suggesting that the fascicles were fatigued in a similar pattern throughout the muscle. The significant reduction of fascicle shortening may reflect an additional strategy to maintain an optimal force output in fatigued conditions, although future work is needed to confirm this notion.


Highly sensitive determination of cyromazine, melamine, and their metabolites in milk by molecularly imprinted solid-phase extraction combined with ultra-performance liquid chromatography.
A novel molecularly imprinted solid-phase extraction-ultra-performance liquid chromatography (MISPE-UPLC) method for effective separation and simultaneous determination of cyromazine, melamine, and their metabolites (ammeline and ammelide) in milk samples was developed. Molecularly imprinted polymers (MIP) were synthesized in an ethanol-water system, with melamine as the template and methacrylic acid as the organic functional monomer. The MIP were applied as a specific sorbent for the selective solid phase extraction of cyromazine, ammelide, melamine and ammeline. The molecular recognition mechanism was investigated by molecular simulation and the experiment was validate by using Fourier transform infrared spectroscopy and (1)H nuclear magnetic resonance spectroscopy. A new mechanism based on the formation of both an amido group and hydrogen bonds was developed. A binding study demonstrated that the MIP showed excellent affinity to and high selectivity for melamine and related compounds. Under optimized conditions, we achieved good linearity of the calibration curves with correlation coefficients >0.999. Low limits of quantification (LOQ) for the method were determined to be 1.25, 1.25, 2.59, and 6.42 µg/kg for cyromazine, ammelide, melamine, and ammeline, respectively, which were 3 orders of magnitude smaller than the maximum residue limit (MRL). The high sensitivity of this method allows detection at the microgram per kilogram level. The proposed MISPE-UPLC method is a highly selective and sensitive method for determination of cyromazine, melamine, and their metabolites (ammeline and ammelide) for use in the control and quality assurance of milk.


Protein phosphorylation in yeast mitochondria: cAMP-dependence, submitochondrial localization and substrates of mitochondrial protein kinases.
We describe the identification and submitochondrial localization of four protein kinases and of their target proteins in derepressed yeast mitochondria. The activity of one of the kinases depends on the presence of cyclic AMP (cAMP). It is soluble and localized in the mitochondrial intermembrane space. Its natural target is a polypeptide of 40 kDa molecular mass, which is bound to the inner membrane. Besides this natural target this kinase phosphorylates acidic heterologous proteins, like casein, with high efficiency. The other protein kinases identified so far are cAMP-independent. At least one is localized in the matrix having its natural substrates (49 and 24 kDa) in the same compartment. Two others are firmly bound to the inner membrane phosphorylating target proteins in the inner membrane (52.5 kDa) and in the intermembrane space (17.5 kDa), respectively.


Cell adhesion molecules P-cadherin and CD24 are markers for carcinoma and dysplasia in the biliary tract.
P-cadherin (CDH3) and CD24 are cell adhesion molecules that control morphogenic processes, cell motility, and invasive growth of tumor cells. The aim of our study was to investigate P-cadherin and CD24 expression in carcinomas and dysplastic lesions of the biliary tract and to evaluate the potential diagnostic usefulness of these cell adhesion molecules. Using immunohistochemistry on tissue microarrays, we analyzed P-cadherin, CD24, and p53 expression in 117 carcinomas of the biliary tract (19 intrahepatic cholangiocarcinomas, 59 extrahepatic cholangiocarcinomas, and 39 gallbladder carcinomas) and correlated our findings with clinicopathologic parameters. We found P-cadherin positivity in 37% of intrahepatic cholangiocarcinomas, 73% of extrahepatic cholangiocarcinomas, and 64% of gallbladder carcinomas, respectively. CD24 reactivity was observed in 21% of intrahepatic cholangiocarcinomas, 58% of extrahepatic cholangiocarcinomas, and 42% of gallbladder carcinomas. Nuclear p53 expression was found in 37% of intrahepatic cholangiocarcinomas, 46% of extrahepatic cholangiocarcinomas, and 45% of gallbladder carcinomas. We also studied P-cadherin, CD24, and p53 expression in normal (n = 30), inflamed (n = 22), and dysplastic (n = 21) biliary epithelium of extrahepatic bile ducts. Dysplastic biliary epithelium was positive for P-cadherin in 91%, for CD24 in 71%, and for p53 in 24% of lesions, respectively. In contrast, normal and inflamed epithelia were negative for all 3 proteins. We conclude that P-cadherin and CD24 are expressed in carcinomas of the biliary tract with high frequency and at an early stage of carcinogenesis. Therefore, they may be useful markers for early detection and as targets for therapy of cholangiocarcinoma.


Tumor cell response to bevacizumab single agent therapy in vitro.
Angiogenesis represents a highly multi-factorial and multi-cellular complex (patho-) physiologic event involving endothelial cells, tumor cells in malignant conditions, as well as bone marrow derived cells and stromal cells. One main driver is vascular endothelial growth factor (VEGFA), which is known to interact with endothelial cells as a survival and mitogenic signal. The role of VEGFA on tumor cells and /or tumor stromal cell interaction is less clear. Condition specific (e.g. hypoxia) or tumor specific expression of VEGFA, VEGF receptors and co-receptors on tumor cells has been reported, in addition to the expression on the endothelium. This suggests a potential paracrine/autocrine loop that could affect changes specific to tumor cells. We used the monoclonal antibody against VEGFA, bevacizumab, in various in vitro experiments using cell lines derived from different tumor entities (non small cell lung cancer (NSCLC), colorectal cancer (CRC), breast cancer (BC) and renal cell carcinoma (RCC)) in order to determine if potential VEGFA signaling could be blocked in tumor cells. The experiments were done under hypoxia, a major inducer of VEGFA and angiogenesis, in an attempt to mimic the physiological tumor condition. Known VEGFA induced endothelial biological responses such as proliferation, migration, survival and gene expression changes were evaluated. Our study was able to demonstrate expression of VEGF receptors on tumor cells as well as hypoxia regulated angiogenic gene expression. In addition, there was a cell line specific effect in tumor cells by VEGFA blockade with bevacizumab in terms of proliferation; however overall, there was a limited measurable consequence of bevacizumab therapy detected by migration and survival. The present study showed in a variety of in vitro experiments with several tumor cell lines from different tumor origins, that by blocking VEGFA with bevacizumab, there was a limited autocrine or cell-autonomous function of VEGFA signaling in tumor cells, when evaluating VEGFA induced downstream outputs known in endothelial cells.


In vitro and transgenic analysis of a human HOXD4 retinoid-responsive enhancer.
Expression of vertebrate Hox genes is regulated by retinoids in cell culture and in early embryonic development. We have identified a 185-bp retinoid-responsive transcriptional enhancer 5' of the human HOXD4 gene, which regulates inducibility of the gene in embryonal carcinoma cells through a pattern of DNA-protein interaction on at least two distinct elements. One of these elements contains a direct repeat mediating ligand-dependent interaction with retinoic acid receptors, and is necessary though not sufficient for the enhancer function. The HOXD4 enhancer directs expression of a lacZ reporter gene in the neural tube of transgenic mouse embryos in a time-regulated and regionally restricted fashion, reproducing part of the anterior neuroectodermal expression pattern of the endogenous Hoxd-4 gene. Administration of retinoic acid to developing embryos causes alterations in the spatial restriction of the transgene expression domain, indicating that the HOXD4 enhancer is also a retinoid-responsive element in vivo. The timing of the retinoic acid response differs from that seen with more 3' Hox genes, in that it occurs much later. This shows that the temporal window of competence in the ability to respond to retinoic acid differs between Hox genes and can be linked to specific enhancers. Mutations in the direct repeat or in a second element in the enhancer affect both retinoid response in culture and developmental regulation in embryos, suggesting that co-operative interactions between different factors mediate the enhancer activity. These data provide further support for a role of endogenous retinoids in regulation and spatial restriction of Hox gene expression in the central nervous system.


Light-dependent hypersensitive response and resistance signaling against Turnip Crinkle Virus in Arabidopsis.
Resistance to Turnip Crinkle Virus (TCV) in Arabidopsis ecotype Dijon (Di)-17 is conferred by the resistance gene HRT and a recessive locus rrt. In Di-17, TCV elicits a hypersensitive response (HR), which is accompanied by increased expression of pathogenesis-related (PR) genes and high levels of salicylic acid (SA). We have previously shown that HRT-mediated resistance to TCV is dependent on SA-mediated signal transduction and that increased levels of SA confer enhanced resistance to TCV via upregulation of the HRT gene. Here we show that HRT-mediated HR and resistance are dependent on light. A dark treatment immediately following TCV inoculation suppressed HR, resistance and activation of the majority of the TCV-induced genes. However, the absence of light did not affect either TCV-induced elevated levels of free SA or the expression of HRT. Interestingly, in the dark, transgenic plants overexpressing HRT showed susceptibility, but overexpression of HRT coupled with high levels of endogenous SA resulted in pronounced resistance. Consistent with these results is the finding that exogenous application of SA prior to TCV inoculation partially overcame the requirement for light. Light was also required for N gene-mediated HR and resistance to Tobacco Mosaic Virus, suggesting that it is an important factor which may be generally required during defense signaling.