Use of Monte Carlo simulation to investigate uncertainty in exposure modeling.
This study used Monte Carlo (MC) simulation to examine the influence of uncertainty on an exposure model and to determine whether a difference exists between two worker groups in a ceramic fiber manufacturing plant. Data on work practices and conditions were gathered in interviews with long-serving employees. With the use of previously developed deterministic modeling techniques and likely distributions for model parameters, MC simulations generated exposure profiles for the two job titles. The exposure profiles overlapped considerably, although the average estimated exposure for one job was approximately double that of the other. However, when the correlation between the model parameters in the two jobs was considered, it was concluded that there was a significant difference in the two estimates. Models are increasingly being used to estimate exposure. Different work situations inevitably result in different exposure estimates. However, it is difficult to determine whether such differences in estimated exposure between worker groups are simply the result of uncertainty with respect to the model parameters or whether they reflect real differences between occupational groups. This study demonstrates the value of MC simulation in helping define the uncertainty in deterministic model estimates.


Brown Adipose Tissue Energy Metabolism in Humans.
The demonstration of metabolically active brown adipose tissue (BAT) in humans primarily using positron emission tomography coupled to computed tomography (PET/CT) with the glucose tracer 18-fluorodeoxyglucose (18FDG) has renewed the interest of the scientific and medical community in the possible role of BAT as a target for the prevention and treatment of obesity and type 2 diabetes (T2D). Here, we offer a comprehensive review of BAT energy metabolism in humans. Considerable advances in methods to measure BAT energy metabolism, including nonesterified fatty acids (NEFA), chylomicron-triglycerides (TG), oxygen, Krebs cycle rate, and intracellular TG have led to very good quantification of energy substrate metabolism per volume of active BAT in vivo. These studies have also shown that intracellular TG are likely the primary energy source of BAT upon activation by cold. Current estimates of BAT's contribution to energy expenditure range at the lower end of what would be potentially clinically relevant if chronically sustained. Yet, 18FDG PET/CT remains the gold-standard defining method to quantify total BAT volume of activity, used to calculate BAT's total energy expenditure. Unfortunately, BAT glucose metabolism better reflects BAT's insulin sensitivity and blood flow. It is now clear that most glucose taken up by BAT does not fuel mitochondrial oxidative metabolism and that BAT glucose uptake can therefore be disconnected from thermogenesis. Furthermore, BAT thermogenesis is efficiently recruited upon repeated cold exposure, doubling to tripling its total oxidative capacity, with reciprocal reduction of muscle thermogenesis. Recent data suggest that total BAT volume may be much larger than the typically observed 50-150 ml with 18FDG PET/CT. Therefore, the current estimates of total BAT thermogenesis, largely relying on total BAT volume using 18FDG PET/CT, may underestimate the true contribution of BAT to total energy expenditure. Quantification of the contribution of BAT to energy expenditure begs for the development of more integrated whole body in vivo methods.


A new phosphate-starvation response in fission yeast requires the endocytic function of myosin I.
Endocytosis is essential for uptake of many substances into the cell, but how it links to nutritional signalling is poorly understood. Here, we show a new role for endocytosis in regulating the response to low phosphate in Schizosaccharomyces pombe. Loss of function of myosin I (Myo1), Sla2/End4 or Arp2, proteins involved in the early steps of endocytosis, led to increased proliferation in low-phosphate medium compared to controls. We show that once cells are deprived of phosphate they undergo a quiescence response that is dependent on the endocytic function of Myo1. Transcriptomic analysis revealed a wide perturbation of gene expression with induction of stress-regulated genes upon phosphate starvation in wild-type but not Δmyo1 cells. Thus, endocytosis plays a pivotal role in mediating the cellular response to nutrients, bridging the external environment and internal molecular functions of the cell.


Global change and marine communities: alien species and climate change.
Anthropogenic influences on the biosphere since the advent of the industrial age are increasingly causing global changes. Climatic change and the rising concentration of greenhouse gases in the atmosphere are ranking high in scientific and public agendas, and other components of global change are also frequently addressed, among which are the introductions of non indigenous species (NIS) in biogeographic regions well separated from the donor region, often followed by spectacular invasions. In the marine environment, both climatic change and spread of alien species have been studied extensively; this review is aimed at examining the main responses of ecosystems to climatic change, taking into account the increasing importance of biological invasions. Some general principles on NIS introductions in the marine environment are recalled, such as the importance of propagule pressure and of development stages during the time course of an invasion. Climatic change is known to affect many ecological properties; it interacts also with NIS in many possible ways. Direct (proximate) effects on individuals and populations of altered physical-chemical conditions are distinguished from indirect effects on emergent properties (species distribution, diversity, and production). Climatically driven changes may affect both local dispersal mechanisms, due to the alteration of current patterns, and competitive interactions between NIS and native species, due to the onset of new thermal optima and/or different carbonate chemistry. As well as latitudinal range expansions of species correlated with changing temperature conditions, and effects on species richness and the correlated extinction of native species, some invasions may provoke multiple effects which involve overall ecosystem functioning (material flow between trophic groups, primary production, relative extent of organic material decomposition, extent of benthic-pelagic coupling). Some examples are given, including a special mention of the situation of the Mediterranean Sea, where so many species have been introduced recently, and where some have spread in very large quantities. An increasing effort by marine scientists is required, not only to monitor the state of the environment, but also to help predicting future changes and finding ways to mitigate or manage them.


The clinical significance of the retrieval of a low number of oocytes following mild ovarian stimulation for IVF: a meta-analysis.
Milder ovarian stimulation protocols for in vitro fertilization (IVF) are being developed to minimize adverse effects. Mild stimulation regimens result in a decreased number of oocytes at retrieval. After conventional ovarian stimulation for IVF, a low number of oocytes are believed to represent poor ovarian reserve resulting in reduced success rates. Recent studies suggest that a similar response following mild stimulation is associated with better outcomes. This review investigates whether the retrieval of a low number of oocytes following mild ovarian stimulation is associated with impaired implantation rates. Three randomized controlled trials comparing the efficacy of the mild ovarian stimulation regimen (involving midfollicular phase initiation of FSH and GnRH co-treatment) for IVF with a conventional long GnRH agonist co-treatment stimulation protocol could be identified by means of a systematic literature search. These studies comprised a total of 592 first treatment cycles. Individual patient data analysis showed that the mild stimulation protocol results in a significant reduction of retrieved oocytes compared with conventional ovarian stimulation (median 6 versus 9, respectively, P < 0.001). Optimal embryo implantation rates were observed with 5 oocytes retrieved following mild stimulation (31%) versus 10 oocytes following conventional stimulation (29%) (P = 0.045). The optimal number of retrieved oocytes depends on the ovarian stimulation regimen. After mild ovarian stimulation, a modest number of oocytes is associated with optimal implantation rates and does not reflect a poor ovarian response. Therefore, the fear of reducing the number of oocytes retrieved following mild ovarian stimulation appears to be unjustified.


A systematic review of chronic disease management interventions in primary care.
Primary and community care are key settings for the effective management of long term conditions. We aimed to evaluate the pattern of health outcomes in chronic disease management interventions for adults with physical health problems implemented in primary or community care settings. The methods were based on our previous review published in 2006. We performed database searches for articles published from 2006 to 2014 and conducted a systematic review with narrative synthesis using the Cochrane Effective Practice and Organisation of Care taxonomy to classify interventions and outcomes. The interventions were mapped to Chronic Care Model elements. The pattern of outcomes related to interventions was summarized by frequency of statistically significant improvements in health care provision and patient outcomes. A total of 9589 journal articles were retrieved from database searches and snowballing. After screening and verification, 165 articles that detailed 157 studies were included. There were few studies with Health Care Organization (1.9% of studies) or Community Resources (0.6% of studies) as the primary intervention element. Self-Management Support interventions (45.8% of studies) most frequently resulted in improvements in patient-level outcomes. Delivery System Design interventions (22.6% of studies) showed benefits in both professional and patient-level outcomes for a narrow range of conditions. Decision Support interventions (21.3% of studies) had impact limited to professional-level outcomes, in particular use of medications. The small number of studies of Clinical Information System interventions (8.9%) showed benefits for both professional- and patient-level outcomes. The published literature has expanded substantially since 2006. This review confirms that Self-Management Support is the most frequent Chronic Care Model intervention that is associated with statistically significant improvements, predominately for diabetes and hypertension.


Immobilized redox mediators for electrochemical NAD(P)+ regeneration.
The applicability of dissolved redox mediators for NAD(P)(+) regeneration has been demonstrated several times. Nevertheless, the use of mediators in solutions for sensor applications is not a very convenient strategy since the analysis is not reagentless and long stabilization times occur. The most important drawbacks of dissolved mediators in biocatalytic applications are interferences during product purification, limited reusability of the mediators, and their cost-intensive elimination from wastewater. Therefore, the use of immobilized mediators has both economic and ecological advantages. This work critically reviews the current state-of-art of immobilized redox mediators for electrochemical NAD(P)(+) regeneration. Various surface modification techniques, such as adsorption polymerization and covalent linkage, as well as the corresponding NAD(P)(+) regeneration rates and the operational stability of the immobilized mediator films, will be discussed. By comparison with other existing regeneration systems, the technical potential and future perspectives of biocatalytic redox reactions based on electrochemically fed immobilized mediators will be assessed.


Open sacrocolpopexy and vaginal apical repair: retrospective comparison of success and serious complications.
We compared treatment success and adverse events between women undergoing open abdominal sacrocolpopexy (ASC) vs vaginal repair (VAR) using data from women enrolled in one of three multicenter trials. We hypothesized that ASC would result in better outcomes than VAR. Participants underwent apical repair of stage 2-4 prolapse. Vaginal repair included uterosacral, sacrospinous, and iliococcygeal suspensions; sacrocolpopexies were via laparotomy. Success was defined as no bothersome bulge symptoms, no prolapse beyond the hymen, and no retreatment up to 24 months. Adverse events were collected at multiple time points. Outcomes were analyzed using longitudinal mixed-effects models to obtain valid outcome estimates at specific visit times, accounting for data missing at random. Comparisons were controlled for center, age, body mass index (BMI), initial Pelvic Organ Prolapse Quantification (POP-Q) stage, baseline scores, prior prolapse repair, and concurrent repairs. Of women who met inclusion criteria (1022 of 1159 eligibile), 701 underwent vaginal repair. The ASC group (n = 321) was older, more likely white, had prior prolapse repairs, and stage 4 prolapse (all p < 0.05). While POP-Q measurements and symptoms improved in both groups, treatment success was higher in the ASC group [odds ratio (OR) 6.00, 95% confidence interval (CI) 3.45-10.44). The groups did not differ significantly in most questionnaire responses at 12 months and overall improvement in bowel and bladder function. By 24 months, fewer patients had undergone retreatment (2% ASC vs 5% VAR); serious adverse events did not differ significantly through 6 weeks (13% vs 5%, OR 2.0, 95% CI 0.9-4.7), and 12 months (26% vs 13%, OR 1.6, 95% CI 0.9-2.9), respectively. Open sacrocolpopexy resulted in more successful prolapse treatment at 2 years.


Cultured smooth muscle targets lack survival activity for ciliary ganglion neurons.
Cultured neurons require specific trophic agents in order to survive. This dependence is thought to resemble the neuron-target interdependence that develops in vivo during synaptogenesis and neuronal cell death. The notion that neurons in general derive trophic support from their synaptic targets is based primarily on studies of peripheral neurons and motor neurons. To assess the general applicability of this nerve-target relationship, we tested the ability of vascular smooth muscle (VSM) to support dissociated neurons from the chick ciliary ganglion. The ciliary ganglion contains 2 distinct neuronal populations, one of which innervates striated muscle, the other VSM. Striated muscle cocultures are known to support all of the neurons in the ganglion for extended periods. Dissociated neurons were therefore cocultured in microwells containing VSM derived from the rat or chick aorta and from the choroid coat of the chick eye. Surviving neurons were counted after 1, 2, 5, and 7 d. Striated muscle is able to support full neuronal survival in the same assay. However, in no case was VSM capable of contributing to neuronal survival in vitro. The neurons in the VSM cocultures were able to form neurites and make contacts with their putative targets, as confirmed by scanning electron and light microscopy. The presence of viable and differentiated smooth muscle cells was demonstrated in the cultures by transmission electron microscopy and analysis of smooth muscle alpha-actin. The failure of VSM and even the choroid target tissue to support the survival of their innervating neurons suggests that novel mechanisms may operate to provide trophic support for neurons innervating VSM targets.


Spreading a medical home redesign: effects on emergency department use and hospital admissions.
The patient-centered medical home (PCMH) is being rapidly deployed in many settings to strengthen US primary care, improve quality, and control costs; however, evidence supporting this transformation is still lacking. We describe the Group Health experience in attempting to replicate the effects on health care use seen in a PCMH prototype clinic via a systemwide spread using Lean as the change strategy. We used an interrupted time series analysis with a patient-month unit of analysis over a 4-year period that included baseline, implementation, and stabilization periods for 412,943 patients. To account for secular trends across these periods, we compared changes in use of face-to-face primary care visits, emergency department visits, and inpatient admissions with those of a nonequivalent comparison group of patients served by community network practices. After accounting for secular trends among network patients, patients empaneled to the PCMH clinics had 5.1% and 6.7% declines in primary care office visits in early and later stabilization years, respectively, after the implementation year. This trend was accompanied by a 123% increase in the use of secure electronic message threads and a 20% increase in telephone encounters. Declines were also seen in emergency department visits at 1 and 2 years (13.7% and 18.5%) compared with what would be expected based on secular trends in network practices. No statistically significant changes were found for hospital admissions. The Group Health experience shows it is possible to reduce emergency department use with PCMH transformation across a diverse set of clinics using a clear change strategy (Lean) and sufficient resources and supports.


Postpartum women in the Honduran health system: folic acid knowledge, attitudes, and practices.
This study had two purposes: first, to determine the knowledge, attitudes, and practices related to folic acid and birth defects among a convenience sample of postpartum Honduran women; and second, to identify food consumption patterns in this population and determine high-consumption staples for potential folic acid fortification. Convenience sampling methodology was used to recruit potential study participants. Participants for this study were 2 619 postpartum Honduran women who had had a normal, in-hospital delivery in one of 16 public hospitals located throughout the country or the two social security hospitals that provide services to the Honduran working class population. Over a 10-month period, in-depth, face-to-face oral interviews, supervised by the research coordinator and staff, were conducted in-hospital prior to discharge. The majority of the women were between 16 and 29 years of age. Approximately half of the respondents (46.4%) had heard of folic acid and over one-third (37.6%) knew that it was a vitamin related to preventing birth defects. Birth defects were most often attributed to drug and alcohol use (20.6%) and lack of vitamin intake (18.1%), but 23.0% related defects to mystical, mythical, or religious causes. Aside from red beans, oranges, and natural fruit juices, folate-rich foods are not widely consumed by this population. The highest consumption frequency of staple foods with the potential to be fortified with folic acid were rice, white flour, corn flour, and pasta. Results from this study provide potential avenues for food fortification, as well as underscore the need for further education about the role of folic acid in the prevention of neural tube defects. Results highlight that standardized health education for Honduran women of reproductive age is needed if folic acid consumption through fortification and supplementation is to be successful and sustainable.


Software comparison for evaluating genomic copy number variation for Affymetrix 6.0 SNP array platform.
Copy number data are routinely being extracted from genome-wide association study chips using a variety of software. We empirically evaluated and compared four freely-available software packages designed for Affymetrix SNP chips to estimate copy number: Affymetrix Power Tools (APT), Aroma.Affymetrix, PennCNV and CRLMM. Our evaluation used 1,418 GENOA samples that were genotyped on the Affymetrix Genome-Wide Human SNP Array 6.0. We compared bias and variance in the locus-level copy number data, the concordance amongst regions of copy number gains/deletions and the false-positive rate amongst deleted segments. APT had median locus-level copy numbers closest to a value of two, whereas PennCNV and Aroma.Affymetrix had the smallest variability associated with the median copy number. Of those evaluated, only PennCNV provides copy number specific quality-control metrics and identified 136 poor CNV samples. Regions of copy number variation (CNV) were detected using the hidden Markov models provided within PennCNV and CRLMM/VanillaIce. PennCNV detected more CNVs than CRLMM/VanillaIce; the median number of CNVs detected per sample was 39 and 30, respectively. PennCNV detected most of the regions that CRLMM/VanillaIce did as well as additional CNV regions. The median concordance between PennCNV and CRLMM/VanillaIce was 47.9% for duplications and 51.5% for deletions. The estimated false-positive rate associated with deletions was similar for PennCNV and CRLMM/VanillaIce. If the objective is to perform statistical tests on the locus-level copy number data, our empirical results suggest that PennCNV or Aroma.Affymetrix is optimal. If the objective is to perform statistical tests on the summarized segmented data then PennCNV would be preferred over CRLMM/VanillaIce. Specifically, PennCNV allows the analyst to estimate locus-level copy number, perform segmentation and evaluate CNV-specific quality-control metrics within a single software package. PennCNV has relatively small bias, small variability and detects more regions while maintaining a similar estimated false-positive rate as CRLMM/VanillaIce. More generally, we advocate that software developers need to provide guidance with respect to evaluating and choosing optimal settings in order to obtain optimal results for an individual dataset. Until such guidance exists, we recommend trying multiple algorithms, evaluating concordance/discordance and subsequently consider the union of regions for downstream association tests.


Synergistic Targeting of the Regulatory and Catalytic Subunits of PI3Kδ in Mature B-cell Malignancies.
Purpose: Aberrant activation of the B-cell receptor (BCR) is implicated in the pathogenesis of mature B-cell tumors, a concept validated in part by the clinical success of inhibitors of the BCR-related kinases BTK (Bruton's tyrosine kinase) and PI3Kδ. These inhibitors have limitations, including the paucity of complete responses, acquired resistance, and toxicity. Here, we examined the mechanism by which the cyclic-AMP/PDE4 signaling axis suppresses PI3K, toward identifying a novel mechanism-based combinatorial strategy to attack BCR-dependency in mature B-cell malignancies.Experimental Design: We used in vitro and in vivo diffuse large B-cell lymphoma (DLBCL) cell lines and primary chronic lymphocytic leukemia (CLL) samples to preclinically evaluate the effects of the combination of the FDA-approved phosphodiesterase 4 (PDE4) inhibitor roflumilast and idelalisib on cell survival and tumor growth. Genetic models of gain- and loss-of-function were used to map multiple signaling intermediaries downstream of the BCR.Results: Roflumilast elevates the intracellular levels of cyclic-AMP and synergizes with idelalisib in suppressing tumor growth and PI3K activity. Mechanistically, we show that roflumilast suppresses PI3K by inhibiting BCR-mediated activation of the P85 regulatory subunit, distinguishing itself from idelalisib, an ATP-competitive inhibitor of the catalytic P110 subunit. Using genetic models, we linked the PDE4-regulated modulation of P85 activation to the oncogenic kinase SYK.Conclusions: These data demonstrate that roflumilast and idelalisib suppress PI3K by distinct mechanisms, explaining the basis for their synergism, and suggest that the repurposing of PDE4 inhibitors to treat BCR-dependent malignancies is warranted. Clin Cancer Res; 24(5); 1103-13. ©2017 AACR.


Detection of 2,4-dinitrotoluene (DNT) as a model system for nitroaromatic compounds via molecularly imprinted short-alkyl-chain SAMs.
A 2-D molecularly imprinted monolayer (2-D MIM) approach was used to prepare a simple and robust sensor for nitroaromatic compounds with 2,4-dinitrotoluene (DNT) as the model compound, which is a precursor and analog for explosive 2,4,6-trinitrotoluene (TNT). In contrast to studies utilizing long-chain hexadecylmercaptan self-assembled monolayers (SAM)s for sensing, a shorter-chain alkylthiol (i.e., butanethiol SAM) was utilized for DNT detection. The role of the chain length of the coadsorbed alkylthiol was emphasized with a matched template during solution adsorption. Semiempirical PM3 quantum calculations were used to determine the molecular conformation and complexation of the adsorbates. A switching mechanism was invoked on the basis of the ability of the template analyte to alter the packing arrangement of the alkylthiol SAMs near defect sites as influenced by the DNT-ethanol solvent complex. A 2-D MIM was formed on the Au surface electrode of a quartz crystal microbalance (QCM), which was then used to sense various concentrations of the analyte. Interestingly, the 2-D MIM QCM also enabled the selective detection of DNT even in a mixed solution of competing molecules, demonstrating the selectivity figure of merit. Likewise, electrochemical impedance spectroscopy (EIS) data at different concentrations of DNT confirmed the 2-D MIM effectiveness for sensing based on the interfacial conformation and electron-transport properties of the imprinted butanethiol SAM.


Cancer as a Social Dysfunction-Why Cancer Research Needs New Thinking.
The incidence and mortality for many cancers continue to rise. As such, critical action is needed on many fronts to reshape how a society thinks, discusses, and fights cancer especially as the population grows and ages. Cancer can be described as a broken social contract that requires different conceptual frameworks such as game theory. To this end, it is our hope that this perspective will catalyze a discussion to rethink the way we approach, communicate, and fund cancer research; thinking of cancer as a broken social contract is only one example. Importantly, this endeavor will require infusion of ideas from other fields such as physics, computational medicine, complexity science, agent-based modeling, sociology, and ecology, all of which have the capacity to drive new insights into cancer biology and clinical medicine. Mol Cancer Res; 16(9); 1346-7. ©2018 AACR.


Neurovascular risks of sacral screws with bicortical purchase: an anatomical study.
The aim of this cadaver study is to define the anatomic structures on anterior sacrum, which are under the risk of injury during bicortical screw application to the S1 and S2 pedicles. Thirty formaldehyde-preserved human male cadavers were studied. Posterior midline incision was performed, and soft tissues and muscles were dissected from the posterior part of the lumbosacral region. A 6 mm pedicle screw was inserted between the superior facet of S1 and the S1 foramen. The entry point of the S2 pedicle screw was located between S1 and S2 foramina. S1 and S2 screws were placed on both right and the left sides of all cadavers. Then, all cadavers were turned into supine position. All abdominal and pelvic organs were moved away and carefully observed for any injury. The tips of the sacral screws were marked and the relations with the anatomic structures were defined. The position of the sacral screws relative to the middle and lateral sacral arteries and veins, and the sacral sympathetic trunk were measured. There was no injury to the visceral organs. In four cases, S1 screw tip was in direct contact with middle sacral artery. In two cases, S1 screw tip was in direct contact with middle sacral vein. It was observed that the S1 screw tips were in close proximity to sacral sympathetic trunk on both right and the left sides. The tip of the S2 screw was in contact with middle sacral artery on the left side only in one case. It is found that the tip of the S2 screw was closely located with the middle sacral vein in two cases. The tip of the S2 pedicle screw was in contact with the sacral sympathetic trunk in eight cases on the right side and seven cases on the left side. Lateral sacral vein was also observed to be disturbed by the S1 and S2 screws. As a conclusion, anterior cortical penetration during sacral screw insertion carries a risk of neurovascular injury. The risk of sacral sympathetic trunk and minor vascular structures together with the major neurovascular structures and viscera should be kept in mind.


Raman spectroscopy of self-catalyzed GaAs(1-x)Sb(x) nanowires grown on silicon.
Thanks to their wide band structure tunability, GaAs(1-x)Sb(x) nanowires provide exciting perspectives in optoelectronic and energy harvesting applications. The control of composition and strain of these ternary alloys is crucial in the determination of their optical and electronic properties. Raman scattering provides information on the vibrational properties of materials, which can be related to the composition and strain. We present a systematic study of the vibrational properties of GaAs(1-x)Sb(x) nanowires for Sb contents from 0 to 44%, as determined by energy-dispersive x-ray analyses. We find that optical phonons red-shift with increasing Sb content. We explain the shift by alloying effects, including mass disorder, dielectric changes and ionic plasmon coupling. The influence of Sb on the surface optical modes is addressed. Finally, we compare the luminescence yield between GaAs and GaAs(1-x)Sb(x), which can be related to a lower surface recombination rate. This work provides a reference for the study of ternary alloys in the form of nanowires, and demonstrates the tunability and high material quality of gold-free ternary antimonide nanowires directly grown on silicon.


Posttranslational modifications, localization, and protein interactions of optineurin, the product of a glaucoma gene.
Glaucoma is a major blinding disease. The most common form of this disease, primary open angle glaucoma (POAG), is genetically heterogeneous. One of the candidate genes, optineurin, is linked principally to normal tension glaucoma, a subtype of POAG. The present study was undertaken to illustrate the basic characteristics of optineurin. Lysates from rat retinal ganglion RGC5 cells were subjected to N- or O-deglycosylation or membrane protein extraction. The phosphorylation status was evaluated after immunoprecipitation. It was found that while phosphorylated, optineurin was neither N- nor O-glycosylated, and was by itself not a membrane protein. RGC5 and human retinal pigment epithelial cells were double stained with anti-optineurin and anti-GM130. The endogenous optineurin exhibited a diffuse, cytoplasmic distribution, but a population of the protein was associated with the Golgi apparatus. Turnover experiments showed that the endogenous optineurin was relatively short-lived, with a half-life of approximately 8 hours. Native blue gel electrophoresis revealed that the endogenous optineurin formed homohexamers. Optineurin also interacted with molecules including Rab8, myosin VI, and transferrin receptor to assemble into supermolecular complexes. When overexpressed, optineurin-green fluorescence protein (GFP) fusion protein formed punctate structures termed "foci" in the perinuclear region. Treatment of nocadazole resulted in dispersion of the optineurin foci. In addition, tetracycline-regulated optineurin-GFPs expressing RGC5 stable cell lines were established for the first time. The present study provides new information regarding basic characteristics of optineurin that are important for future efforts in defining precisely how optineurin functions normally and how mutations may result in pathology. The inducible optineurin-GFP-expressing cell lines are also anticipated to facilitate in-depth studies of optineurin. Furthermore, the demonstrations that optineurin is an aggregation-prone protein and that the foci formation is microtubule-dependent bear similarities to features documented in neurodegenerative diseases, supporting a neurodegenerative paradigm for glaucoma.


Intraspecific sequence comparisons reveal similar rates of non-collinear gene insertion in the B and D genomes of bread wheat.
Polyploidization is considered one of the main mechanisms of plant genome evolution. The presence of multiple copies of the same gene reduces selection pressure and permits sub-functionalization and neo-functionalization leading to plant diversification, adaptation and speciation. In bread wheat, polyploidization and the prevalence of transposable elements resulted in massive gene duplication and movement. As a result, the number of genes which are non-collinear to genomes of related species seems markedly increased in wheat. We used new-generation sequencing (NGS) to generate sequence of a Mb-sized region from wheat chromosome arm 3DS. Sequence assembly of 24 BAC clones resulted in two scaffolds of 1,264,820 and 333,768 bases. The sequence was annotated and compared to the homoeologous region on wheat chromosome 3B and orthologous loci of Brachypodium distachyon and rice. Among 39 coding sequences in the 3DS scaffolds, 32 have a homoeolog on chromosome 3B. In contrast, only fifteen and fourteen orthologs were identified in the corresponding regions in rice and Brachypodium, respectively. Interestingly, five pseudogenes were identified among the non-collinear coding sequences at the 3B locus, while none was found at the 3DS locus. Direct comparison of two Mb-sized regions of the B and D genomes of bread wheat revealed similar rates of non-collinear gene insertion in both genomes with a majority of gene duplications occurring before their divergence. Relatively low proportion of pseudogenes was identified among non-collinear coding sequences. Our data suggest that the pseudogenes did not originate from insertion of non-functional copies, but were formed later during the evolution of hexaploid wheat. Some evidence was found for gene erosion along the B genome locus.


Work and health among immigrants and native Swedes 1990-2008: a register-based study on hospitalization for common potentially work-related disorders, disability pension and mortality.
There are many immigrants in the Swedish workforce, but knowledge of their general and work-related health is limited. The aim of this register-based study was to explore whether documented migrant residents in Sweden have a different health status regarding receipt of a disability pension, mortality and hospitalization for lung, heart, psychiatric, and musculoskeletal disorders compared with the native population, and if there were variations in relation to sex, geographical origin, position on the labor market, and time since first immigration. This study included migrants to Sweden since 1960 who were 28-47 years old in 1990, and included 243 860 individuals. The comparison group comprised a random sample of 859 653 native Swedes. These cohorts were followed from 1991 to 2008 in national registers. The immigrants were divided into four groups based on geographic origin. Hazard ratios for men and women from different geographic origins and with different employment status were analyzed separately for the six outcomes, with adjustment for age, education level, and income. The influence of length of residence in Sweden was analyzed separately. Nordic immigrants had increased risks for all investigated outcomes while most other groups had equal or lower risks for those outcomes than the Swedes. The lowest HRs were found in the EU 15+ group (from western Europe, North America, Australia and New Zealand). All groups, except Nordic immigrants, had lower risk of mortality, but all had higher risk of disability pension receipt compared with native Swedes. Unemployed non-Nordic men displayed equal or lower HRs for most outcomes, except disability pension receipt, compared with unemployed Swedish men. A longer time since first immigration improved the health status of men, while women showed opposite results. Employment status and length of residence are important factors for health. The contradictory results of low mortality and high disability pension risks need more attention. There is great potential to increase the knowledge in this field in Sweden, because of the high quality registers.


Identification of genetic risk for pancreatic adenocarcinoma.
Recent consortium guidelines support research-based screening for those at high risk of pancreatic cancer (pancreatic ductal adenocarcinoma (PDAC)). Genetic testing plays an important role in the establishment of high-risk PDAC research clinics by delineating those individuals who would benefit from screening protocols. We retrospectively examined patients referred for PDAC-related genetic testing from January 2009 to June 2014. Patients were referred for a personal and/or family history of PDAC or a questioned diagnosis of hereditary pancreatitis (HP). Of the 75 referred patients, 36 underwent testing, of which 11 (31%) were mutation-positive. In total, 36% of patients with chronic pancreatitis carried a mutation, 11% of patients with a family history of PDAC carried a mutation, and 20% of patients with a personal history of PDAC carried a mutation. The most common barrier to testing was lack of insurance coverage. Genetic testing yields a suitable number of mutation-positive individuals who may benefit from increased screening. Subjects with possible HP yielded the highest positive rate. Individuals with idiopathic pancreatitis, onset of pancreatitis before the age of 30 years, and those with a family history of PDAC should be considered for testing. Sub-optimal insurance coverage remains a major deterrent to obtaining testing.


Highly pathogenic avian influenza virus subtype H5N1 in Africa: a comprehensive phylogenetic analysis and molecular characterization of isolates.
Highly pathogenic avian influenza virus A/H5N1 was first officially reported in Africa in early 2006. Since the first outbreak in Nigeria, this virus spread rapidly to other African countries. From its emergence to early 2008, 11 African countries experienced A/H5N1 outbreaks in poultry and human cases were also reported in three of these countries. At present, little is known of the epidemiology and molecular evolution of A/H5N1 viruses in Africa. We have generated 494 full gene sequences from 67 African isolates and applied molecular analysis tools to a total of 1,152 A/H5N1 sequences obtained from viruses isolated in Africa, Europe and the Middle East between 2006 and early 2008. Detailed phylogenetic analyses of the 8 gene viral segments confirmed that 3 distinct sublineages were introduced, which have persisted and spread across the continent over this 2-year period. Additionally, our molecular epidemiological studies highlighted the association between genetic clustering and area of origin in a majority of cases. Molecular signatures unique to strains isolated in selected areas also gave us a clearer picture of the spread of A/H5N1 viruses across the continent. Mutations described as typical of human influenza viruses in the genes coding for internal proteins or associated with host adaptation and increased resistance to antiviral drugs have also been detected in the genes coding for transmembrane proteins. These findings raise concern for the possible human health risk presented by viruses with these genetic properties and highlight the need for increased efforts to monitor the evolution of A/H5N1 viruses across the African continent. They further stress how imperative it is to implement sustainable control strategies to improve animal and public health at a global level.


Impact of Neurosurgery Medical Student Research Grants on Neurosurgery Residency Choice.
Recent decades have seen a rapid expansion of involvement of medical students in biomedical research during medical school training. Research within medical school has been shown to influence medical students with regard to medical knowledge, career development, and residency specialty choice. The objective of this study was to evaluate the impact of neurosurgery medical student research grants on neurosurgery residency choice and provide an insight on the demographics of grant awardees. In this retrospective study, a search of award recipients was performed using data available on the American Association of Neurological Surgeons, Congress of Neurological Surgeons, and Neurosurgery Research and Education Foundation websites. Searched years included the first cycle of American Association of Neurological Surgeons/Neurosurgery Research and Education Foundation (2007) and Council of State Neurosurgical Societies/Congress of Neurological Surgeons (2008-2009) grant awards until the 2015-2016 cycle, which is the latest award cycle to date. The initial search yielded 163 research grants that were awarded to 158 students between the years of 2007 and 2016. Among the 163 grant recipients, 126 (77.3%) were men. Among the 88 recipients who entered postgraduate residency programs, 51% (45 of 88) matched into neurosurgery residency. When considering both neurosurgery and neurology residency programs, the percentage increased to 59.1% (52 of 88). Neurosurgery grants for medical students are highly successful in producing future neurosurgeons with >50% of grant recipients matched into neurosurgery. Women are underrepresented in neurosurgery grants and neurosurgery residency programs. This situation can be improved by providing insight about the field early in medical school, perhaps through increased use of neurosurgery medical student grants.


Walsby's square bacterium: fine structure of an orthogonal procaryote.
The "square" bacterium, first described by Walsby from brine collected at the Red Sea shore [A. E. Walsby, Nature (London) 283:69-71, 1980] was examined by electron microscopy. The cells appeared as flat rectangular boxes in scanning electron micrographs. In sections and freeze-fracture preparation, the edges looked more rounded. The thickness apparently remains constant as the cells grow and divide. Their sides were a few micrometers long, but the cells were only 0.25 micrometers thick. They showed typical procaryote structure, with a regular cell wall and a gas vacuole fine structure similar to that of other halophilic procaryotes. The inner fracture faces of the cell membrane showed a much denser population of intramembrane particles than the outer fracture faces, but no patches of purple membrane, despite the presence of bacteriorhodospin-like pigment in the cell suspension. Morphologically identical cells have been found in brine from Baja California, Mexico.


Innovative legal approaches to address obesity.
The law is a powerful public health tool with considerable potential to address the obesity issue. Scientific advances, gaps in the current regulatory environment, and new ways of conceptualizing rights and responsibilities offer a foundation for legal innovation. This article connects developments in public health and nutrition with legal advances to define promising avenues for preventing obesity through the application of the law. Two sets of approaches are defined: (1) direct application of the law to factors known to contribute to obesity and (2) original and innovative legal solutions that address the weak regulatory stance of government and the ineffectiveness of existing policies used to control obesity. Specific legal strategies are discussed for limiting children's food marketing, confronting the potential addictive properties of food, compelling industry speech, increasing government speech, regulating conduct, using tort litigation, applying nuisance law as a litigation strategy, and considering performance-based regulation as an alternative to typical regulatory actions. Finally, preemption is an overriding issue and can play both a facilitative and a hindering role in obesity policy. Legal solutions are immediately available to the government to address obesity and should be considered at the federal, state, and local levels. New and innovative legal solutions represent opportunities to take the law in creative directions and to link legal, nutrition, and public health communities in constructive ways.


Morphological priming in Spanish verb forms: an ERP repetition priming study.
The ERP repetition priming paradigm has been shown to be sensitive to the processing differences between regular and irregular verb forms in English and German. The purpose of the present study is to extend this research to a language with a different inflectional system, Spanish. The design (delayed visual repetition priming) was adopted from our previous study on English, and the specific linguistic phenomena we examined are priming relations between different kinds of stem (or root) forms. There were two experimental conditions: In the first condition, the prime and the target shared the same stem form, e.g., "ando-andar" [I walk-to walk], whereas in the second condition, the prime contained a marked (alternated) stem, e.g., "duermo-dormir" [I sleep-to sleep]. A reduced N400 was found for unmarked (nonalternated) stems in the primed condition, whereas marked stems showed no such effect. Moreover, control conditions demonstrated that the surface form properties (i.e., the different degree of phonetic and orthographic overlap between primes and targets) do not explain the observed priming difference. The ERP priming effect for verb forms with unmarked stems in Spanish is parallel to that found for regularly inflected verb forms in English and German. We argue that effective priming is possible because prime target pairs such as "ando-andar" access the same lexical entry for their stems. By contrast, verb forms with alternated stems (e.g., "duermo") constitute separate lexical entries, and are therefore less powerful primes for their corresponding base forms.


Theileria infection in domestic ruminants in northern Ethiopia.
Piroplasmosis caused by different tick-borne hemoprotozoan parasites of the genera Theileria and Babesia is among the most economically important infections of domestic ruminants in sub-Saharan Africa. A survey for piroplasm infection was conducted in three locations in Northern Ethiopia. Of 525 domestic ruminants surveyed, 80% of the cattle, 94% of the sheep and 2% of the goats were positive for different Theileria spp. based on PCR of blood followed by DNA sequencing. Sheep had a significantly higher rate of infection compared with cattle (P<0.0003) and both sheep and cattle had higher rates of infection compared to goats (P<0.0001). Four species of Theileria were detected in cattle: T. velifera, T. mutans, T. orientalis complex and T. annulata with infection rates of 66, 8, 4, and 2%, respectively. This is the first report of T. annulata, the cause of Tropical Theileriosis in Ethiopia. Of the two Theileria spp. detected in small ruminants, T. ovis was highly prevalent (92%) in sheep and rare in goats (1.5%) whereas T. seperata was infrequent in sheep (2%) and rare in goats (0.4%). None of the animals were positive for Babesia spp.; however, Sarcocystis capracanis and S. tenella were detected in one goat and a sheep, respectively. The widespread distribution of Theileria spp. among cattle in northern Ethiopia including the virulent T. annulata and more mildly pathogenic T. mutans and T. orientalis, and the high infection rate in sheep with the usually sub-clinical T. ovis indicate extensive exposure to ticks and transmission of piroplasms with an important economic impact.


Characterization of the Ornithine Hydroxylation Step in Albachelin Biosynthesis.
N-Hydroxylating monooxygenases (NMOs) are involved in siderophore biosynthesis. Siderophores are high affinity iron chelators composed of catechol and hydroxamate functional groups that are synthesized and secreted by microorganisms and plants. Recently, a new siderophore named albachelin was isolated from a culture of Amycolatopsis alba growing under iron-limiting conditions. This work focuses on the expression, purification, and characterization of the NMO, abachelin monooxygenase (AMO) from A. alba. This enzyme was purified and characterized in its holo (FAD-bound) and apo (FAD-free) forms. The apo-AMO could be reconstituted by addition of free FAD. The two forms of AMO hydroxylate ornithine, while lysine increases oxidase activity but is not hydroxylated and display low affinity for NADPH.


The nonpsychoactive cannabis constituent cannabidiol is an oral anti-arthritic therapeutic in murine collagen-induced arthritis.
The therapeutic potential of cannabidiol (CBD), the major nonpsychoactive component of cannabis, was explored in murine collagen-induced arthritis (CIA). CIA was elicited by immunizing DBA/1 mice with type II collagen (CII) in complete Freund's adjuvant. The CII used was either bovine or murine, resulting in classical acute CIA or in chronic relapsing CIA, respectively. CBD was administered after onset of clinical symptoms, and in both models of arthritis the treatment effectively blocked progression of arthritis. CBD was equally effective when administered i.p. or orally. The dose dependency showed a bell-shaped curve, with an optimal effect at 5 mg/kg per day i.p. or 25 mg/kg per day orally. Clinical improvement was associated with protection of the joints against severe damage. Ex vivo, draining lymph node cells from CBD-treated mice showed a diminished CII-specific proliferation and IFN-gamma production, as well as a decreased release of tumor necrosis factor by knee synovial cells. In vitro effects of CBD included a dose-dependent suppression of lymphocyte proliferation, both mitogen-stimulated and antigen-specific, and the blockade of the Zymosan-triggered reactive oxygen burst by peritoneal granulocytes. It also was found that CBD administration was capable of blocking the lipopolysaccharide-induced rise in serum tumor necrosis factor in C57/BL mice. Taken together, these data show that CBD, through its combined immunosuppressive and anti-inflammatory actions, has a potent anti-arthritic effect in CIA.


Diagnosis of Upper and Lower Respiratory Tract Bacterial Infections with the Use of Multiplex PCR Assays.
The investigation of respiratory infections by molecular techniques provides important information about the epidemiology of respiratory disease, especially during the post-vaccination era. The objective of the present study was the detection of bacterial pathogens directly in clinical samples from patients with upper and lower respiratory tract infections using multiplex polymerase chain reaction (PCR) assays developed in our laboratory. Clinical samples taken over a three-year period (2007-2009) and obtained from 349 patients (adults (n = 66); children (n = 283)) with signs and symptoms of certain upper or lower respiratory tract infections, consisted of: bronchoalveolar lavages (BAL, n = 83), pleural fluids (n = 29), and middle-ear aspirates (n = 237). Overall, 212 samples (61%) were confirmed by culture and/or PCR. Among the positive samples, Streptococcus pneumoniae (mainly serotype 3) was predominant (104/212; 49.0%), followed by non-typable Haemophilus influenzae (NTHi) 59/212; 27.8%) and Streptococcus pyogenes (47/212; 22%). Haemophilus influenzae type b was detected in only three samples. The underlying microbiology of respiratory infections is gradually changing in response to various selective pressures, such as vaccine use and antibiotic consumption. The application of multiplex PCR (mPCR) assays is particularly useful since it successfully identified the microorganisms implicated in acute otitis media or lower respiratory tract infections in nearly 75% of patients with a positive result compared to conventional cultures. Non-culture identification of the implicated pneumococcal serotypes is also an important issue for monitoring pneumococcal infections in the era of conjugate pneumococcal vaccines.


Identification of proliferative and mature β-cells in the islets of Langerhans.
Insulin-dependent diabetes is a complex multifactorial disorder characterized by loss or dysfunction of β-cells. Pancreatic β-cells differ in size, glucose responsiveness, insulin secretion and precursor cell potential; understanding the mechanisms that underlie this functional heterogeneity might make it possible to develop new regenerative approaches. Here we show that Fltp (also known as Flattop and Cfap126), a Wnt/planar cell polarity (PCP) effector and reporter gene acts as a marker gene that subdivides endocrine cells into two subpopulations and distinguishes proliferation-competent from mature β-cells with distinct molecular, physiological and ultrastructural features. Genetic lineage tracing revealed that endocrine subpopulations from Fltp-negative and -positive lineages react differently to physiological and pathological changes. The expression of Fltp increases when endocrine cells cluster together to form polarized and mature 3D islet mini-organs. We show that 3D architecture and Wnt/PCP ligands are sufficient to trigger β-cell maturation. By contrast, the Wnt/PCP effector Fltp is not necessary for β-cell development, proliferation or maturation. We conclude that 3D architecture and Wnt/PCP signalling underlie functional β-cell heterogeneity and induce β-cell maturation. The identification of Fltp as a marker for endocrine subpopulations sheds light on the molecular underpinnings of islet cell heterogeneity and plasticity and might enable targeting of endocrine subpopulations for the regeneration of functional β-cell mass in diabetic patients.


Transfer of Minibeam Radiation Therapy into a cost-effective equipment for radiobiological studies: a proof of concept.
Minibeam radiation therapy (MBRT) is an innovative synchrotron radiotherapy technique able to shift the normal tissue complication probability curves to significantly higher doses. However, its exploration was hindered due to the limited and expensive beamtime at synchrotrons. The aim of this work was to develop a cost-effective equipment to perform systematic radiobiological studies in view of MBRT. Tumor control for various tumor entities will be addressable as well as studies to unravel the distinct biological mechanisms involved in normal and tumor tissues responses when applying MBRT. With that aim, a series of modifications of a small animal irradiator were performed to make it suitable for MBRT experiments. In addition, the brains of two groups of rats were irradiated. Half of the animals received a standard irradiation, the other half, MBRT. The animals were followed-up for 6.5 months. Substantial brain damage was observed in the group receiving standard RT, in contrast to the MBRT group, where no significant lesions were observed. This work proves the feasibility of the transfer of MBRT outside synchrotron sources towards a small animal irradiator.


Splitting a DNAzyme enables a Na+-dependent FRET signal from the embedded aptamer.
Recently, a few Na+-specific RNA-cleaving DNAzymes have been reported, and a Na+ aptamer was identified from the NaA43 and Ce13d DNAzymes. These DNAzymes and the embedded aptamer have been used for Na+ detection. In this work, we studied the Na+-dependent folding of the Ce13d DNAzyme using fluorescence resonance energy transfer (FRET). When a FRET donor and an acceptor were respectively labeled at the ends of the DNAzyme, Na+ failed to induce an obvious end-to-end distance change, suggesting a rigid global structure. To relax this rigidity, the Ce13d DNAzyme was systematically split at various sites on both the enzyme and the substrate strands. The Na+ binding activity of the split structures was characterized by 2-aminopurine fluorescence, enzymatic activity, Tb3+-sensitized luminescence, and DMS footprinting. Among the various constructs, the only one that retained Na+ binding was the split at the cleavage site, and this construct was further labeled with two dyes near the split site. This FRET result showed Na+-dependent folding with a Kd of 26 mM Na+. This study provides important structural information related to Na+ binding to this new aptamer, which might also be useful for future work in biosensor design.


Effective Detection of Porcine Cytomegalovirus Using Non-Invasively Taken Samples from Piglets.
Shortage of human organs forced the development of xenotransplantation using cells, tissues, and organs from pigs. Xenotransplantation may be associated with the transmission of porcine zoonotic microorganisms, among them the porcine cytomegalovirus (PCMV). To prevent virus transmission, pigs have to be screened using sensitive methods. In order to perform regular follow-ups and further breeding of the animals, samples for testing should be collected by low-invasive or non-invasive methods. Sera, ear biopsies, as well as oral and anal swabs were collected from ten 10-day-old Aachen minipigs (AaMP) and tested for PCMV using sensitive nested polymerase chain reaction (PCR) as well as uniplex and duplex real-time PCR. Porcine cytomegalovirus DNA was detected most frequently in oral and anal swabs. Comparison of duplex and uniplex real-time PCR systems for PCMV detection demonstrated a lower sensitivity of duplex real-time PCR when the copy numbers of the target genes were low (less 200). Therefore, to increase the efficacy of PCMV detection in piglets, early testing of oral and anal swabs by uniplex real-time PCR is recommended.


Genome-Wide Analysis of Ocular Adnexal Lymphoproliferative Disorders Using High-Resolution Single Nucleotide Polymorphism Array.
We identified the genomic signature of ocular adnexal lymphoproliferative disorders (LPDs), especially ocular adnexal mucosa-associated lymphoid tissue (MALT) lymphoma, IgG4-related ophthalmic disease (IgG4-ROD), reactive lymphoid hyperplasia (RLH), and diffuse large B-cell lymphoma (DLBCL). We included 52 subjects with ocular adnexal LPDs (13 orbital MALT lymphomas, 16 conjunctival MALT lymphomas, 13 IgG4-RODs, 4 RLHs, and 6 DLBCLs) who had been treated at the Tokyo Medical University Hospital from 2008 to 2012. Genomic DNA was extracted from the tumor tissues and subjected to high-resolution single nucleotide polymorphism array (SNP-A) karyotyping using GeneChip Human Mapping 250K SNP arrays. The array data were investigated using Copy Number Analysis for GeneChips (CNAG) software. In ocular adnexal MALT lymphomas, the most frequent copy number (CN) gain region was trisomy 3 detected in 31% (9/29), followed by trisomy 18 in 17% (5/29), and 6p and 21q in 14% (4/29). The most frequent CN loss regions were 6q and 9p, detected in 7% (2/29). Uniparental disomy was detected on 6q in 14% (4/29), followed by 3q in 10% (3/29). Copy number variations (CNVs) were not detected in IgG4-RODs and RLHs. Conversely, CNVs were more frequent in DLBCLs than in ocular adnexal MALT lymphomas. Copy number variations were detected in 77% (10/13) of orbital MALT lymphomas and in 67% (11/16) of conjunctival MALT lymphomas. High-resolution single nucleotide polymorphism array is a useful method for discriminating ocular adnexal lymphomas from benign LPDs. The differences in the chromosomal abnormality patterns may reflect the activity of ocular adnexal LPDs.


Lactobacillus delbrueckii ssp. bulgaricus B-30892 can inhibit cytotoxic effects and adhesion of pathogenic Clostridium difficile to Caco-2 cells.
Probiotic microorganisms are receiving increasing interest for use in the prevention, treatment, or dietary management of certain diseases, including antibiotic-associated diarrhea (AAD). Clostridium difficile is the most common cause of AAD and the resulting C. difficile - mediated infection (CDI), is potentially deadly. C. difficile associated diarrhea (CDAD) is manifested by severe inflammation and colitis, mostly due to the release of two exotoxins by C. difficile causing destruction of epithelial cells in the intestine. The aim of this study was to determine the effect of probiotic bacteria Lactobacillus delbrueckii ssp. bulgaricus B-30892 (LDB B-30892) on C. difficile-mediated cytotoxicity using Caco-2 cells as a model. Experiments were carried out to test if the cytotoxicity induced by C. difficile-conditioned-medium on Caco-2 cells can be altered by cell-free supernatant (CFS) from LDB B-30892 in different dilutions (1:2 to 1:2048). In a similar experimental setup, comparative evaluations of other probiotic strains were made by contrasting the results from these strains with the results from LDB B-30892, specifically the ability to affect C. difficile induced cytotoxicity on Caco-2 monolayers. Adhesion assays followed by quantitative analysis by Giemsa staining were conducted to test if the CFSs from LDB B-30892 and other probiotic test strains have the capability to alter the adhesion of C. difficile to the Caco-2 monolayer. Experiments were also performed to evaluate if LDB B-30892 or its released components have any bactericidal effect on C. difficile. Co-culturing of LDB B-30892 with C. difficile inhibited the C. difficile-mediated cytotoxicity on Caco-2 cells. When CFS from LDB B-30892-C. difficile co-culture was administered (up to a dilution of 1:16) on Caco-2 monolayer, there were no signs of cytotoxicity. When CFS from separately grown LDB B-30892 was mixed with the cell-free toxin preparation (CFT) of separately cultured C. difficile, the LDB B-30892 CFS was inhibitory to C. difficile CFT-mediated cytotoxicity at a ratio of 1:8 (LDB B-30892 CFS:C. difficile CFT). We failed to find any similar inhibition of C. difficile-mediated cytotoxicity when other probiotic organisms were tested in parallel to LDB B-30892. Our data of cytotoxicity experiments suggest that LDB B-30892 releases one or more bioactive component(s) into the CFS, which neutralizes the cytotoxicity induced by C. difficile, probably by inactivating its toxin(s). Our data also indicate that CFS from LDB B-30892 reduced the adhesion of C. difficile by 81%, which is significantly (P <0.01) higher than all other probiotic organisms tested in this study. This study reveals the very first findings that Lactobacillus delbrueckii ssp. bulgaricus B-30892 (LDB B-30892) can eliminate C. difficile-mediated cytotoxicity, using Caco-2 cells as a model. The study also demonstrates that LDB B-30892 can reduce the colonization of C. difficile cells in colorectal cells. More study is warranted to elucidate the specific mechanism of action of such reduction of cytotoxicity and colonization.


Intake of dietary soy isoflavones in relation to perimenstrual symptoms of Korean women living in the USA.
This study was conducted to identify the potential relationship between the dietary intake level of soy isoflavones and perimenstrual symptomatology. The research design was a cross-sectional study. The sample was made up of 84 Korean women living in the USA, aged 28-40 years. The Moos Menstrual Distress Questionnaire (MDQ) and the Food Frequency Questionnaire were used as measurement tools. The soy isoflavone intake was significantly correlated with MDQ scores in the menstrual phase. In conclusion, the beneficial effect of dietary soy isoflavones on certain menstrual symptoms was established, suggesting that soy isoflavones could be one of the dietary factors related to the complexity of premenstrual syndrome (PMS). The positive effect of soy isoflavones on PMS warrants further study.


Reference values for muscle strength: a systematic review with a descriptive meta-analysis.
Muscle strength is an important component of health. To describe and evaluate the studies which have established the reference values for muscle strength on healthy individuals and to synthesize these values with a descriptive meta-analysis approach. A systematic review was performed in MEDLINE, LILACS, and SciELO databases. Studies that investigated the reference values for muscle strength of two or more appendicular/axial muscle groups of health individuals were included. Methodological quality, including risk of bias was assessed by the QUADAS-2. Data extracted included: country of the study, sample size, population characteristics, equipment/method used, and muscle groups evaluated. Of the 414 studies identified, 46 were included. Most of the studies had adequate methodological quality. Included studies evaluated: appendicular (80.4%) and axial (36.9%) muscles; adults (78.3%), elderly (58.7%), adolescents (43.5%), children (23.9%); isometric (91.3%) and isokinetic (17.4%) strength. Six studies (13%) with similar procedures were synthesized with meta-analysis. Generally, the coefficient of variation values that resulted from the meta-analysis ranged from 20.1% to 30% and were similar to those reported by the original studies. The meta-analysis synthesized the reference values of isometric strength of 14 muscle groups of the dominant/non-dominant sides of the upper/lower limbs of adults/elderly from developed countries, using dynamometers/myometer. Most of the included studies had adequate methodological quality. The meta-analysis provided reference values for the isometric strength of 14 appendicular muscle groups of the dominant/non-dominant sides, measured with dynamometers/myometers, of men/women, of adults/elderly. These data may be used to interpret the results of the evaluations and establish appropriate treatment goals.


Fibrosis as an end point for clinical trials in liver disease: a report of the international fibrosis group.
Deaths due to the consequences of advanced liver fibrosis and cirrhosis remain a significant cause of mortality worldwide. Biologically plausible pathways involved in hepatic fibrogenesis have also led to the identification of numerous preclinical and clinical compounds that have received great interest as potential future therapeutic agents for patients with liver fibrosis. With this in mind, stake holders from academia, regulatory agencies, clinicians, and the pharmaceutical industry met to understand and discuss the many complex issues involved in developing potential therapeutic agents which act primarily through modifying fibrosis and to discuss appropriate end points for clinical trials in these patient populations. In this article, we summarize those discussions and attempt to highlight many of the hurdles and unanswered questions as we attempt to move forward and develop therapies to combat liver fibrosis.


Turing and Wolpert work together during limb development.
Limb development has been widely studied in developmental biology. Two types of theories, one based on positional information and another based on spontaneous pattern formation, are utilized to understand this process. However, for historical reasons, these two concepts are regarded as mutually exclusive. By considering the interaction between these two mechanisms with quantitative morphological data, we can experimentally explore how the vertebrate limb skeletal pattern is established from a new direction.


A monoclonal antibody recognizing very late activation antigen-4 inhibits eosinophil accumulation in vivo.
Using an in vivo test system, the role of the beta 1 integrin very late activation antigen-4 (VLA-4) in eosinophil accumulation in allergic and nonallergic inflammatory reactions was investigated. Eosinophil infiltration and edema formation were measured as the local accumulation of intravenously injected 111In-labeled eosinophils and 125I-human serum albumin. The inflammatory reactions investigated were a passive cutaneous anaphylaxis (PCA) reaction and responses elicited by intradermal soluble inflammatory mediators (platelet-activating factor, leukotriene B4, C5a des Arg), arachidonic acid, and zymosan particles. The in vitro pretreatment of 111In-eosinophils with the anti-VLA-4 monoclonal antibody (mAb) HP1/2, which crossreacts with guinea pig eosinophils, suppressed eosinophil accumulation in all the inflammatory reactions investigated. Eosinophil accumulation was inhibited to the same extent when mAb HP1/2 was administered intravenously. It is interesting that HP1/2 had no effect on stimulated edema formation. These results suggest a role for VLA-4 in eosinophil accumulation in vivo and indicate a dissociation between the inflammatory events of eosinophil accumulation and edema formation.


Neural tube and neural crest: a new view with time-lapse high-definition photomicroscopy.
The dynamic process of neural tube formation and neural crest migration in live, unstained cultured avian embryos at Hamburger-Hamilton (H.H.) stages 8-11 was investigated by time-lapse cinematography using a high-definition microscope. These studies have demonstrated that neural tube closure in the trunk region differs from that observed in the head. The cephalic neural folds elevate slowly, then make contact rapidly. Following this initial apposition, they gradually "zip-up" in the rostrad and caudad direction. In the trunk region where the neuroepithelium bulges adjacent to the somites, the edges of the folds pulsate and forcefully touch-retract-touch in these bulging regions; the intersomitic epithelia retract, remain open even after more posterior somitic regions have apposed, and then close slowly. Epithelial blebs and N-CAM antibody were observed at the leading edges of the neuroepithelia. Between the open folds only a few bridging cells were seen; they probably represent the sites of initial cell adhesion following epithelial retraction. Focusing into the developing embryo shows that neuroepithelial fusion occurs prior to surface epithelial fusion. A meshwork of synchronously pulsating neural crest cells was identified below the surface epithelium and a preliminary investigation of their initial migration was conducted.


On language.
Abstract Steven Pinker is a professor in the Department of Brain and Cognitive Sciences at MIT, and in 1994 will become director of its McDonnell-Pew Center for Cognitive Neuroscience. He received his B.K from McGill University in 1976 and his Ph.D. from Harvard University in 1979, both in experimental psychology, and taught at Harvard and Stanford before joining the faculty of MIT in 1982. He has done research in visual cognition and the psychology of language, and is the author of Language Learnability and Language Development (1984) and Learnability and Cognition (1989) and the editor of Visual Cognition (1985), Connections and Symbol (1988, with Jacques Mehler), and Lexical and Conceptual Semantics (1992, with Beth Levin). He was the recipient of the Early Career Award in 1984 and the Boyd McCandless Award in 1986 from the American Psychological Association, a Graduate Teaching Award from MIT in 1986, and the Troland Research Award from the National Academy of Sciences in 1993. His newest book, The Language Instinct, will be published by William Morrow & Company in January 1994.


Transcription of the human U2 snRNA genes continues beyond the 3' box in vivo.
The 3' box of the human class II snRNA genes is required for proper 3' processing of transcripts, but how it functions is unclear. Several lines of evidence suggest that termination of transcription occurs at the 3' box and the terminated transcript is then a substrate for processing. However, using nuclear run-on analysis of endogenous genes, we demonstrate that transcription continues for at least 250 nucleotides beyond the 3' box of the U2 genes. Although in vivo footprinting analysis of both the U1 and U2 genes detects no protein-DNA contacts directly over the 3' box, a series of G residues immediately downstream from the 3' box of the U1 gene are clearly protected from methylation by dimethylsulfate. In conjunction with the 3' box of the U1 gene, this in vivo footprinted region causes termination of transcription of transiently transfected U2 constructs, whereas a 3' box alone does not. Taken together, these results indicate that the 3' box is not an efficient transcriptional terminator but may act as a processing element that is functional in the nascent RNA.


The combination of the preoperative albumin-bilirubin grade and the fibrosis-4 index predicts the prognosis of patients with hepatocellular carcinoma after liver resection.
There is little information regarding the use of a combination of the albumin-bilirubin (ALBI) grade and the fibrosis-4 index (FIB-4) in predicting hepatocellular carcinoma (HCC) patient outcomes after liver resection. In this study, we aimed to analyze the predictive ability of a combination of the ALBI grade and the FIB-4 score (ALBI-FIB-4) for HCC patients within the Milan criteria after liver resection. The data of HCC patients within the Milan criteria who underwent liver resection between 2011 and 2019 at our center were reviewed (n = 544). Patients with an FIB-4 index > 3.25 were considered to have a high FIB-4 index and were given a score of 1, whereas patients with an FIB-4 index ≤ 3.25 were considered to have a low FIB-4 index and were given a score of 0. The ALBI-FIB-4 score was a summary score that combined the ALBI grade and the score based on the FIB-4 index. During the follow-up period, 279 patients experienced recurrence, and 175 patients died. Multivariate analysis showed that tumor size, the presence of multiple tumors, the presence of microvascular invasion and the ALBI-FIB-4 score were four independent risk factors for both postoperative recurrence-free survival (RFS) and overall survival (OS). The 5-year RFS of patients with high ALBI-FIB-4 scores of 1, 2, and 3 were 55.0%, 44.2% and 35.3%, respectively (p = 0.004). The 5-year OS rates of patients with high ALBI-FIB-4 scores of 1, 2, and 3 were 72.9%, 66.4% and 54.8%, respectively (p = 0.011). The ALBI-FIB-4 score may be a surrogate marker for predicting the prognosis of patients with HCC after liver resection. A high ALBI-FIB-4 score was associated with a high incidence of postoperative recurrence and mortality.


Multiple osteochondromas of the antlers and cranium in a free-ranging white-tailed deer (Odocoileus virginianus).
This paper reports a case of multiple osteochondromas affecting the antlers and the left zygomatic bone of a free-ranging adult white-tailed buck (Odocoileus virginianus) from Georgia, USA. Along with a few postcranial bones, the antlered cranium of the individual was found in a severely weathered condition and devoid of any soft tissue. The antlers exhibited five pedunculated exostoses that were composed of cancellous bone and, in their peripheral portions, also mineralized cartilage. The largest of the exostoses, located on the right antler, had a maximum circumference of 55 cm. The exostosis arising from the zygomatic bone was broad-based and much smaller than the exophytic outgrowths on the antlers. Diagnosis of the exostoses as osteochondromas was based on their overall morphology, the normal bone structure in their stalk regions, and the continuity of their spongiosa and cortex with the respective components of the parent bones. Antleromas, i.e., pathological outgrowths developing on antlers as a result of insufficient androgen production, were excluded in the differential diagnosis, based on (1) the apparent maturity and, except for the tumors, normal shape of the antlers and (2) the fact that exostosis formation had also affected the zygomatic bone. Previously only a single case of solitary osteochondroma of an antler has been described in the scientific literature. The case presented here is the first report of multiple osteochondromas in a deer. As antlers are regularly collected as trophies, and huge numbers of them are critically inspected each year, the fact that thus far only two cases of antler osteochondromas have been reported suggests that these tumors are very rare.


Solar ultraviolet radiation induces biological alterations in human skin in vitro: relevance of a well-balanced UVA/UVB protection.
Cutaneous damages such as sunburn, pigmentation, and photoaging are known to be induced by acute as well as repetitive sun exposure. Not only for basic research, but also for the design of the most efficient photoprotection, it is crucial to understand and identify the early biological events occurring after ultraviolet (UV) exposure. Reconstructed human skin models provide excellent and reliable in vitro tools to study the UV-induced alterations of the different skin cell types, keratinocytes, fibroblasts, and melanocytes in a dose- and time-dependent manner. Using different in vitro human skin models, the effects of UV light (UVB and UVA) were investigated. UVB-induced damages are essentially epidermal, with the typical sunburn cells and DNA lesions, whereas UVA radiation-induced damages are mostly located within the dermal compartment. Pigmentation can also be obtained after solar simulated radiation exposure of pigmented reconstructed skin model. Those models are also highly adequate to assess the potential of sunscreens to protect the skin from UV-associated damage, sunburn reaction, photoaging, and pigmentation. The results showed that an effective photoprotection is provided by broad-spectrum sunscreens with a potent absorption in both UVB and UVA ranges.


[Are we prepared to diagnose and managed malignant hyperthermia?].
Malignant Hyperthermia (MH) is an uncommon but potentially lethal disease associated to halogenate agents and/or succinylcholine exposure. Resulting mortality and morbidity may be decreased by early diagnosis and specific treatment, including sodium dantrolene. Brazil has approximately 160 million inhabitants assisted by more than 6000 anesthesiologists. In the last decade, special attention was given to this disease, resulting in better informed anesthesiologists and more prepared hospitals to treat malignant hyperthermia (MH). This study aimed at measuring the level of information of Brazilian anesthesiologists regarding MH, in order to develop new approaches to control this disease. A questionnaire with 20 questions on malignant hyperthermia diagnosis, prevention and treatment was mailed to the 6,164 members of the Brazilian Society of Anesthesiology. Questionnaires were returned by 646 anesthesiologists (10.4%). More than 90% of correct answers about clinical diagnosis and treatment were obtained. On the other hand, nearly 50% of anesthesiologists gave incorrect answers about muscle biopsy indication and clinical pharmacology of dantrolene. Results have indicated adequate knowledge about this topic, but has shown that some relevant issues need additional attention. The number of answers was significant to evaluate MH understanding of Brazilian anesthesiologists, but has also shown poor motivation. From these results it is possible to conclude that it is essential to intensify continuing education programs, contemplating all issues of this major anesthetic subject.


Clinical end points and response criteria in mycosis fungoides and Sézary syndrome: a consensus statement of the International Society for Cutaneous Lymphomas, the United States Cutaneous Lymphoma Consortium, and the Cutaneous Lymphoma Task Force of the European Organisation for Research and Treatment of Cancer.
Mycosis fungoides (MF) and Sézary syndrome (SS), the major forms of cutaneous T-cell lymphoma, have unique characteristics that distinguish them from other types of non-Hodgkin's lymphomas. Clinical trials in MF/SS have suffered from a lack of standardization in evaluation, staging, assessment, end points, and response criteria. Recently defined criteria for the diagnosis of early MF, guidelines for initial evaluation, and revised staging and classification criteria for MF and SS now offer the potential for uniform staging of patients enrolled in clinical trials for MF/SS. This article presents consensus recommendations for the general conduct of clinical trials of patients with MF/SS as well as methods for standardized assessment of potential disease manifestations in skin, lymph nodes, blood, and visceral organs, and definition of end points and response criteria. These guidelines should facilitate collaboration among investigators and collation of data from sponsor-generated or investigator-initiated clinical trials involving patients with MF or SS.


Common data elements for clinical research in mitochondrial disease: a National Institute for Neurological Disorders and Stroke project.
The common data elements (CDE) project was developed by the National Institute of Neurological Disorders and Stroke (NINDS) to provide clinical researchers with tools to improve data quality and allow for harmonization of data collected in different research studies. CDEs have been created for several neurological diseases; the aim of this project was to develop CDEs specifically curated for mitochondrial disease (Mito) to enhance clinical research. Nine working groups (WGs), composed of international mitochondrial disease experts, provided recommendations for Mito clinical research. They initially reviewed existing NINDS CDEs and instruments, and developed new data elements or instruments when needed. Recommendations were organized, internally reviewed by the Mito WGs, and posted online for external public comment for a period of eight weeks. The final version was again reviewed by all WGs and the NINDS CDE team prior to posting for public use. The NINDS Mito CDEs and supporting documents are publicly available on the NINDS CDE website ( https://commondataelements.ninds.nih.gov/ ), organized into domain categories such as Participant/Subject Characteristics, Assessments, and Examinations. We developed a comprehensive set of CDE recommendations, data definitions, case report forms (CRFs), and guidelines for use in Mito clinical research. The widespread use of CDEs is intended to enhance Mito clinical research endeavors, including natural history studies, clinical trial design, and data sharing. Ongoing international collaboration will facilitate regular review, updates and online publication of Mito CDEs, and support improved consistency of data collection and reporting.


The Anolis lizard genome: an amniote genome without isochores.
Isochores are large regions of relatively homogeneous nucleotide composition and are present in the genomes of all mammals and birds that have been sequenced to date. The newly sequenced genome of Anolis carolinensis provides the first opportunity to quantify isochore structure in a nonavian reptile. We find Anolis to have the most compositionally homogeneous genome of all amniotes sequenced thus far, a homogeneity exceeding that for the frog Xenopus. Based on a Bayesian algorithm, Anolis has smaller and less GC-rich isochores compared with human and chicken. Correlates generally associated with GC-rich isochores, including shorter introns and higher gene density, have all but disappeared from the Anolis genome. Using genic GC as a proxy for isochore structure so as to compare with other vertebrates, we found that GC content has substantially decreased in the lineage leading to Anolis since diverging from the common ancestor of Reptilia ∼275 Ma, perhaps reflecting weakened or reversed GC-biased gene conversion, a nonadaptive substitution process that is thought to be important in the maintenance and trajectory of isochore evolution. Our results demonstrate that GC composition in Anolis is not associated with important features of genome structure, including gene density and intron size, in contrast to patterns seen in mammal and bird genomes.


[Primary care in the USA and the Peruvian experience in perspective].
Due to a complex payment system, the health system of the United States is fragmented, expensive, and achieves the goal of improving the health of the entire population. Excessive health costs increase the budget deficit in the US. This situation makes to the government to have greater willingness to try new approaches in the delivery of health services. Is well know that the states that have a solid system of primary health care (PC), spent less money in diseases and have lower utilization of health services (reflected in fewer days of hospitalization). This is why the AP is spreading in the US making doctors, nurses and other providers to collaborate on new proposals for team-based care, coordinated and patient-oriented generating real benefits while controlling costs. The US experience with AP support evidence-based approach to propose, from a foreign perspective, strengthening the AP in Peru.


Isolation of Hendra virus from pteropid bats: a natural reservoir of Hendra virus.
Since it was first described in Australia in 1994, Hendra virus (HeV) has caused two outbreaks of fatal disease in horses and humans, and an isolated fatal horse case. Our preliminary studies revealed a high prevalence of neutralizing antibodies to HeV in bats of the genus PTEROPUS:, but it was unclear whether this was due to infection with HeV or a related virus. We developed the hypothesis that HeV excretion from bats might be related to the birthing process and we targeted the reproductive tract for virus isolation. Three virus isolates were obtained from the uterine fluid and a pool of foetal lung and liver from one grey-headed flying-fox (Pteropus poliocephalus), and from the foetal lung of one black flying-fox (P. alecto). Antigenically, these isolates appeared to be closely related to HeV, returning positive results on immunofluorescent antibody staining and constant-serum varying-virus neutralization tests. Using an HeV-specific oligonucleotide primer pair, genomic sequences of the isolates were amplified. Sequencing of 200 nucleotides in the matrix gene identified that these three isolates were identical to HeV. Isolations were confirmed after RNA extracted from original material was positive for HeV RNA when screened on an HeV Taqman assay. The isolation of HeV from pteropid bats corroborates our earlier serological and epidemiological evidence that they are a natural reservoir host of the virus.


Sonic hedgehog is expressed in epithelial cells during development of whisker, hair, and tooth.
Sonic hedgehog (Shh) is a vertebrate gene homologous to a Drosophila segment polarity gene, hedgehog, and functions as a secreted signaling molecule in limb pattern formation, differentiation of motor neurons, and sclerotome induction. We found that Shh is also expressed in epithelia of the developing whisker, hair, tooth, rugae, and thyroid primordium of mouse embryos. In whisker and hair development, Shh is expressed in epithelial cells before condensation of the underlying mesenchymal cells and then in the placode. The expression of Shh continues in the hair bulb surrounding the dermal papilla. Shh is also expressed in epithelial cells of the tooth bud, then localized to the enamel knot. The Shh expression continues in developing ameloblasts. These results suggest that SHH is an essential epithelial signaling molecule in epithelio-mesenchymal interactions for the terminal differentiation.


Comparison between periodic and stochastic parabolic light trapping structures for thin-film microcrystalline Silicon solar cells.
Light trapping is of very high importance for silicon photovoltaics (PV) and especially for thin-film silicon solar cells. In this paper we investigate and compare theoretically the light trapping properties of periodic and stochastic structures having similar geometrical features. The theoretical investigations are based on the actual surface geometry of a scattering structure, characterized by an atomic force microscope. This structure is used for light trapping in thin-film microcrystalline silicon solar cells. Very good agreement is found in a first comparison between simulation and experimental results. The geometrical parameters of the stochastic structure are varied and it is found that the light trapping mainly depends on the aspect ratio (length/height). Furthermore, the maximum possible light trapping with this kind of stochastic structure geometry is investigated. In a second step, the stochastic structure is analysed and typical geometrical features are extracted, which are then arranged in a periodic structure. Investigating the light trapping properties of the periodic structure, we find that it performs very similar to the stochastic structure, in agreement with reports in literature. From the obtained results we conclude that a potential advantage of periodic structures for PV applications will very likely not be found in the absorption enhancement in the solar cell material. However, uniformity and higher definition in production of these structures can lead to potential improvements concerning electrical characteristics and parasitic absorption, e.g. in a back reflector.


Treatment of child anxiety disorders via guided parent-delivered cognitive-behavioural therapy: randomised controlled trial.
Promising evidence has emerged of clinical gains using guided self-help cognitive-behavioural therapy (CBT) for child anxiety and by involving parents in treatment; however, the efficacy of guided parent-delivered CBT has not been systematically evaluated in UK primary and secondary settings. To evaluate the efficacy of low-intensity guided parent-delivered CBT treatments for children with anxiety disorders. A total of 194 children presenting with a current anxiety disorder, whose primary carer did not meet criteria for a current anxiety disorder, were randomly allocated to full guided parent-delivered CBT (four face-to-face and four telephone sessions) or brief guided parent-delivered CBT (two face-to-face and two telephone sessions), or a wait-list control group (trial registration: ISRCTN92977593). Presence and severity of child primary anxiety disorder (Anxiety Disorders Interview Schedule for DSM-IV, child/parent versions), improvement in child presentation of anxiety (Clinical Global Impression - Improvement scale), and change in child anxiety symptoms (Spence Children's Anxiety Scale, child/parent version and Child Anxiety Impact scale, parent version) were assessed at post-treatment and for those in the two active treatment groups, 6 months post-treatment. Full guided parent-delivered CBT produced superior diagnostic outcomes compared with wait-list at post-treatment, whereas brief guided parent-delivered CBT did not: at post-treatment, 25 (50%) of those in the full guided CBT group had recovered from their primary diagnosis, compared with 16 (25%) of those on the wait-list (relative risk (RR) 1.85, 95% CI 1.14-2.99); and in the brief guided CBT group, 18 participants (39%) had recovered from their primary diagnosis post-treatment (RR = 1.56, 95% CI 0.89-2.74). Level of therapist training and experience was unrelated to child outcome. Full guided parent-delivered CBT is an effective and inexpensive first-line treatment for child anxiety.


Whole body exposure with GSM 900MHz affects spatial memory in mice.
Extended work has been performed worldwide on the effects of mobile phone radiation upon rats' cognitive functions, however there is great controversy to the existence or not of deficits. The present work has been designed in order to test the effects of mobile phone radiation on spatial learning and memory in mice Mus musculus Balb/c using the Morris water maze (a hippocampal-dependent spatial memory task), since there is just one other study on mice with very low SAR level (0.05W/kg) showing no effects. We have applied a 2h daily dose of pulsed GSM 900MHz radiation from commercially available mobile phone for 4 days at SAR values ranging from 0.41 to 0.98W/kg. Statistical analysis revealed that during learning, exposed animals showed a deficit in transferring the acquired spatial information across training days (increased escape latency and distance swam, compared to the sham-exposed animals, on the first trial of training days 2-4). Moreover, during the memory probe-trial sham-exposed animals showed the expected preference for the target quadrant, while the exposed animals showed no preference, indicating that the exposed mice had deficits in consolidation and/or retrieval of the learned spatial information. Our results provide a basis for more thorough investigations considering reports on non-thermal effects of electromagnetic fields (EMFs).


The impact of exercise training on adipose tissue remodelling in cancer cachexia.
Cachexia affects the majority of patients with advanced cancer and no effective treatment is currently available to address this paraneoplastic syndrome. It is characterized by a reduction in body weight due to the loss of white adipose tissue (WAT) and skeletal muscle. The loss of WAT seems to occur at an earlier time point than skeletal muscle proteolysis, with recent evidence suggesting that the browning of WAT may be a major contributor to this process. Several factors seem to modulate WAT browning including pro-inflammatory cytokines; however, the underlying molecular pathways are poorly characterized. Exercise training is currently recommended for the clinical management of low-grade inflammatory conditions as cancer cachexia. While it seems to counterbalance the impairment of skeletal muscle function and attenuate the loss of muscle mass, little is known regarding its effects in adipose tissue. The browning of WAT is one of the mechanisms through which exercise improves body composition in overweight/obese individuals. While this effect is obviously advantageous in this clinical setting, it remains to be clarified if exercise training could protect or exacerbate the cachexia-related catabolic phenotype occurring in adipose tissue of cancer patients. Herein, we overview the molecular players involved in adipose tissue remodelling in cancer cachexia and in exercise training and hypothesize on the mechanisms modulated by the synergetic effect of these conditions. A better understanding of how physical activity regulates body composition will certainly help in the development of successful multimodal therapeutic strategies for the clinical management of cancer cachexia.


Use of the circular Dammann grating in angle measurement.
We describe a novel method of angle measurement by borrowing the concept of the circular Dammann grating (CDG). A three-order CDG is employed in this experiment. The displacement of the tilted angle can be determined accurately by measuring the projection from the distorted CDG image. This method is controlled only by the initial radius of the image and the converging ratio of the lens. Compared with conventional techniques, this technique has the advantages of a simple design with superior resolution to within 1 degree, low cost, and compactness. A theoretical analysis together with experimental results is presented.


Coalescent inference for infectious disease: meta-analysis of hepatitis C.
Genetic analysis of pathogen genomes is a powerful approach to investigating the population dynamics and epidemic history of infectious diseases. However, the theoretical underpinnings of the most widely used, coalescent methods have been questioned, casting doubt on their interpretation. The aim of this study is to develop robust population genetic inference for compartmental models in epidemiology. Using a general approach based on the theory of metapopulations, we derive coalescent models under susceptible-infectious (SI), susceptible-infectious-susceptible (SIS) and susceptible-infectious-recovered (SIR) dynamics. We show that exponential and logistic growth models are equivalent to SI and SIS models, respectively, when co-infection is negligible. Implementing SI, SIS and SIR models in BEAST, we conduct a meta-analysis of hepatitis C epidemics, and show that we can directly estimate the basic reproductive number (R(0)) and prevalence under SIR dynamics. We find that differences in genetic diversity between epidemics can be explained by differences in underlying epidemiology (age of the epidemic and local population density) and viral subtype. Model comparison reveals SIR dynamics in three globally restricted epidemics, but most are better fit by the simpler SI dynamics. In summary, metapopulation models provide a general and practical framework for integrating epidemiology and population genetics for the purposes of joint inference.


Toenail selenium as biomarker: reproducibility over a one-year period and factors influencing reproducibility.
We assessed the reproducibility of selenium levels in toenails, comparing concentrations in two sets of specimens collected about a year apart, from 80 women (40 pre-menopausal and 40 post-menopausal) in the period October 1990 to February 1992. The women were participants in a prospective study on hormones and diet in relation to prediagnostic breast cancer (the ORDET study) conducted in northern Italy. Toenail selenium was determined by electrothermal atomic absorption spectrometry. The data were log-transformed as they were not normally distributed. To assess reproducibility Pearson correlation coefficients (r) for the two selenium determinations were calculated in pre- and post-menopausal women, according to smoking status and acetone treatment. A weighted kappa statistic (k) assessed inter-quintile agreement between the two sets of measures. Toenail selenium levels were highly reproducible (r = 0.57, p < 0.001), especially in pre-menopausal women (r = 0.66, p <0.001). Smoking lowered selenium levels (mean difference of 0.24 microg/g, p < 0.05 between smokers and non smokers) but did not significantly influence reproducibility. Acetone treatment to remove nail polish did not modify selenium levels but affected inter-quintile agreement, with moderate agreement (k = 0.58, p < 0.001) when acetone was used at both or neither samplings; and fair non significant agreement (k = 0.39, p = 0.06) when acetone was used at one sampling but not the other. As selenium levels in toenails are highly reproducible, notwithstanding variation in selenium levels in food and long-term changes in individuals' food choices, toenail selenium may be a useful biomarker of selenium exposure, particularly since toenail samples provide a measure of long-term exposure. However age, smoking status and acetone treatment are possible causes of misclassification.