Paxillin and ponsin interact in nascent costameres of muscle cells.
Muscle differentiation requires the transition from motile myoblasts to sessile myotubes and the assembly of a highly regular contractile apparatus. This striking cytoskeletal remodelling is coordinated with a transformation of focal adhesion-like cell-matrix contacts into costameres. To assess mechanisms underlying this differentiation process, we searched for muscle specific-binding partners of paxillin. We identified an interaction of paxillin with the vinexin adaptor protein family member ponsin in nascent costameres during muscle differentiation, which is mediated by an interaction of the second src homology domain 3 (SH3) domain of ponsin with the proline-rich region of paxillin. To understand the molecular basis of this interaction, we determined the structure of this SH3 domain at 0.83 A resolution, as well as its complex with the paxillin binding peptide at 1.63 A resolution. Upon binding, the paxillin peptide adopts a polyproline-II helix conformation in the complex. Contrary to the charged SH3 binding interface, the peptide contains only non-polar residues and for the first time such an interaction was observed structurally in SH3 domains. Fluorescence titration confirmed the ponsin/paxillin interaction, characterising it further by a weak binding affinity. Transfection experiments revealed further characteristics of ponsin functions in muscle cells: All three SH3 domains in the C terminus of ponsin appeared to synergise in targeting the protein to force-transducing structures. The overexpression of ponsin resulted in altered muscle cell-matrix contact morphology, suggesting its involvement in the establishment of mature costameres. Further evidence for the role of ponsin in the maintenance of mature mechanotransduction sites in cardiomyocytes comes from the observation that ponsin expression was down-regulated in end-stage failing hearts, and that this effect was reverted upon mechanical unloading. These results provide new insights in how low affinity protein-protein interactions may contribute to a fine tuning of cytoskeletal remodelling processes during muscle differentiation and in adult cardiomyocytes.


Transrectal electrical impedance tomography of the prostate: spatially coregistered pathological findings for prostate cancer detection.
Prostate cancer ranks as one of the most common malignancies and currently represents the second leading cancer-specific cause of death in men. The current use of single modality transrectal ultrasound (TRUS) for biopsy guidance has a limited sensitivity and specificity for accurately identifying cancerous lesions within the prostate. This study introduces a novel prostate cancer imaging method that combines TRUS with electrical impedance tomography (EIT) and reports on initial clinical findings based on in vivo measurements. The ultrasound system provides anatomic information, which guides EIT image reconstruction. EIT reconstructions are correlated with semiquantitative pathological findings. Thin plate spline warping transformations are employed to overlay electrical impedance images and pathological maps describing the spatial distribution of prostate cancer, with the latter used as reference for data analysis. Clinical data were recorded from a total of 50 men prior to them undergoing radical prostatectomy for prostate cancer treatment. Student's t-tests were employed to statistically examine the electrical property difference between cancerous tissue and benign tissue as defined through histological assessment of the excised gland. Example EIT reconstructions are presented along with a statistical analysis comparing EIT and pathology. An average transformation error of 1.67% is found when 381 spatially coregistered pathological images are compared with their target EIT reconstructed counterparts. At EIT signal frequencies of 0.4, 3.2, and 25.6 kHz, paired-testing demonstrated that the conductivity of cancerous regions is significantly greater than that of benign regions ( p < 0.0304). These preliminary clinical findings suggest the potential benefits electrical impedance measurements might have for prostate cancer detection.


Developmental changes of FOXP3-expressing CD4+CD25+ regulatory T cells and their impairment in patients with FOXP3 gene mutations.
FOXP3 is required for the generation and function of CD4(+)CD25(+) regulatory T (Treg) cells. To elucidate the biological role of Treg cells, we used a monoclonal anti-FOXP3 antibody to examine the frequencies of Treg cells during child development. The percentages of CD4(+)CD25(+)FOXP3(+) T cells were constant shortly from after birth through adulthood. CD4(+)CD25(+)FOXP3(+) T cells in cord blood showed the naive CD45RA(+)CD45RO(-) phenotype, whereas adult CD4(+)CD25(+)FOXP3(+) T cells expressed mostly the memory CD45RA(-)CD45RO(+) phenotype. The age-dependent dominance of memory CD4(+)CD25(+)FOXP3(+) T cells implies functional differences between naive and memory Treg cells. Notably, four patients with FOXP3 gene mutations revealed a paucity of CD4(+)CD25(+)FOXP3(+) T cells. Importantly, one patient with a frame shift mutation, who showed typical symptoms of IPEX (immune dysregulation, polyendocrinopathy, enteropathy, X-linked), exhibited marked T cell activation, whereas others with missense mutations, who were clinically milder, did not. This observation suggests a possible genotype-phenotype correlation in IPEX.


Prospective patient stratification into robust cancer-cell intrinsic subtypes from colorectal cancer biopsies.
Colorectal cancer (CRC) biopsies underpin accurate diagnosis, but are also relevant for patient stratification in molecularly-guided clinical trials. The consensus molecular subtypes (CMSs) and colorectal cancer intrinsic subtypes (CRISs) transcriptional signatures have potential clinical utility for improving prognostic/predictive patient assignment. However, their ability to provide robust classification, particularly in pretreatment biopsies from multiple regions or at different time points, remains untested. In this study, we undertook a comprehensive assessment of the robustness of CRC transcriptional signatures, including CRIS and CMS, using a range of tumour sampling methodologies currently employed in clinical and translational research. These include analyses using (i) laser-capture microdissected CRC tissue, (ii) eight publically available rectal cancer biopsy data sets (n = 543), (iii) serial biopsies (from AXEBeam trial, NCT00828672; n = 10), (iv) multi-regional biopsies from colon tumours (n = 29 biopsies, n = 7 tumours), and (v) pretreatment biopsies from the phase II rectal cancer trial COPERNCIUS (NCT01263171; n = 44). Compared to previous results obtained using CRC resection material, we demonstrate that CMS classification in biopsy tissue is significantly less capable of reliably classifying patient subtype (43% unknown in biopsy versus 13% unknown in resections, p = 0.0001). In contrast, there was no significant difference in classification rate between biopsies and resections when using the CRIS classifier. Additionally, we demonstrated that CRIS provides significantly better spatially- and temporally- robust classification of molecular subtypes in CRC primary tumour tissue compared to CMS (p = 0.003 and p = 0.02, respectively). These findings have potential to inform ongoing biopsy-based patient stratification in CRC, enabling robust and stable assignment of patients into clinically-informative arms of prospective multi-arm, multi-stage clinical trials. © 2018 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland.


Novel dry cryotherapy system for cooling the equine digit.
Digital cryotherapy is commonly used for laminitis prophylaxis and treatment. Currently validated methods for distal limb cryotherapy involve wet application or compression technology. There is a need for a practical, affordable, dry cryotherapy method that effectively cools the digit. The objective of this study was to evaluate the hoof wall surface temperatures (HWSTs) achieved with a novel dry cryotherapy technology. Repeated-measures in vivo experimental study. Experimental intervention at a single site. 6 systemically healthy horses (3 mares, 3 geldings). Cryotherapy was applied to six horses for eight hours with a commercially available rubber and rubber and welded fabricice boot, which extended proximally to include the foot and pastern. Reusable malleable cold therapy packs were secured against the foot and pastern with the three built-in hook-and-loop fastener panels. HWST and pastern surface temperature of the cryotherapy-treated limb, HWST of the control limb and ambient temperature were recorded every five minutes throughout the study period. Results were analysed with mixed-effects multivariable regression analysis. The HWST (median 11.1°C, interquartile range 8.6°C-14.7°C) in the cryotherapy-treated limb was significantly decreased compared with the control limb (median 29.7°C, interquartile range 28.9°C-30.4°C) (P≤0.001). Cryotherapy limb HWST reached a minimum of 6.75°C (median) with an interquartile range of 4.1°C-9.3°C. Minimum HWST was achieved 68 minutes after cryotherapy pack application. Dry application of cryotherapy significantly reduced HWST and reached minimums below the therapeutic target of 10°C. This cryotherapy method might offer an effective alternative for digital cooling.


Effectiveness of icing as a postharvest treatment for control of Vibrio vulnificus and Vibrio parahaemolyticus in the eastern oyster (Crassostrea virginica).
The focus of this research was to investigate the efficacy of icing as a postharvest treatment for reduction of the levels of Vibrio vulnificus and Vibrio parahaemolyticus in commercial quantities of shellstock oysters. The experiments were conducted in June and August of 2006 and consisted of the following treatments: (i) on-board icing immediately after harvest; (ii) dockside icing approximately 1 to 2 h prior to shipment; and (iii) no icing (control). Changes in the levels of pathogenic Vibrio spp. during wholesale and retail handling for 2 weeks postharvest were also monitored. On-board icing achieved temperature reductions in all sacks in accordance with the National Shellfish Sanitation Program standard, but dockside icing did not meet this standard. Based on one-way analysis of variance, the only statistically significant relationship between Vibrio levels and treatment occurred for samples harvested in August; in this case, the levels of V. vulnificus in the noniced oysters were significantly higher (P < 0.05) than were the levels in the samples iced on-board. When analyzing counts over the 14-day storage period, using factorial analysis, there were statistically significant differences in V. vulnificus and V. parahaemolyticus levels by sample date and/or treatment (P < 0.05), but these relationships were not consistent. Treated (iced) oysters had significantly higher gaping (approximately 20%) after 1 week in cold storage than did noniced oysters (approximately 10%) and gaping increased significantly by day 14 of commercial storage. On-board and dockside icing did not predictably reduce the levels of V. vulnificus or V. parahaemolyticus in oysters, and icing negatively impacted oyster survival during subsequent cold storage.


Comparison of the clinical and laboratory presentations of primary and secondary glomerular diseases.
To investigate the clinicopathologic correlation of the glomerular diseases and to compare the presentations of primary and secondary glomerular diseases. Two hundred and sixty adult patients (men = 56.2%, mean age = 39.9 ± 16.0 years) who had undergone renal biopsy and been diagnosed with glomerular disease were retrospectively investigated. The information about the demographic and clinical characteristics of the patients, the indications for biopsy, the laboratory parameters, and the pathological results of the biopsies were obtained from the medical records. Primary and secondary glomerulonephritis (GN) groups were compared in terms of their demographic characteristics, indications for biopsy, and laboratory parameters. Percent of patients with primary and secondary GN were 63.5% and 36.5%, respectively. The most common indication for biopsy was nephrotic syndrome (53.8%) in all patients. It was found that presentation with impaired renal function was more prevalent in the secondary GN group when compared with the primary GN group (54.7% vs. 35.8%, p < 0.05). Among the inflammation parameters, the levels of erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) were found to be significantly higher in the secondary GN group when compared with the primary GN group (ESR = 85.2 ± 30.4 mm/h vs. 56.3 ± 30.7 mm/h, p < 0.001; CRP = 42.1 ± 39.6 mg/L vs. 21.7 ± 40.2 mg/L, p < 0.001). The clinicopathologic correlation of GN was found similar with the larger studies in the literature. This study also indicated that the rate of patients with secondary GN was not low, and the inflammation parameters were higher in the secondary GN groups.


A cost analysis of single-stage bilateral versus two-stage direct anterior total hip arthroplasty.
The potential cost savings of single-stage bilateral total hip arthroplasty (THA) are unclear, and the risks associated with it are not well defined. We sought to compare the costs and perioperative complications of single-stage bilateral THA via the direct anterior approach (DAA) to a two-stage bilateral protocol. We retrospectively reviewed patients who underwent a single- stage bilateral DAA THA and compared them to a two-stage THA group. We conducted a cost analysis from both the hospital perspective and the Ministry of Health (MOH) perspective. 24 patients were included in this study. The 2 groups were similar in age (58.9 vs 63.9 yrs), height (169.2 vs 170.9 cm), weight (80.2 vs 78.6 kg), BMI (27.9 vs 26.3 kg/m2), ASA score (2.2 vs 2.2), and CCI score (2.3 vs 2.9). The mean cost per patient from the hospital perspective for the single-stage group was $10,728.13 (SD = 621.46) compared to $12,670.63 (SD = 519.72) for the two-stage group (Mean Difference = $1,942.50, 95% CI = $1,457.49 to $2,427.51, p<0.001). Similarly, from the MOH perspective, the cost for the single-stage group was $12,552.34 (SD = 644.93) compared to $14,740.58 (SD = 598.07) for the two-stage group (Mean Difference = $2,188.24, 95% CI = $1,661.67 to $2,714.81, p<0.001). There were no significant differences in complication rate between groups. The largest percent of total cost savings from a hospital perspective was attributed to cost of operating room staff and OR set-up (55%). Our results suggest that single-stage bilateral DAA THA results in significant cost savings compared to two-stage DAA THA.


Ethambutol dosage for the treatment of children: literature review and recommendations.
The currently recommended daily dose of ethambutol (EMB) for the treatment of tuberculosis (TB) in children varies from a maximum daily dose of 15 mg/kg body weight daily (without a range) to 15-20 mg/kg and 20 mg/ kg (range 15-25 mg/kg). Published evidence relating to the dosage, toxicity and pharmacokinetics of EMB in children and adults is reviewed and a dose of EMB for use in childhood is recommended. Using key words 'ethambutol', 'childhood', 'TB', 'pharmacokinetics', 'bioavailability' and 'toxicity', Medline searches were conducted; cross-references were sought from original papers, books and conference proceedings dating from 1961. When English summaries were available, data were extracted from papers in languages other than English. EMB has a dose-related efficacy best seen when given to adults alone or with a single other drug. Together with isoniazid (INH), a dose of 15 mg/kg EMB gave better results than 6 mg/kg, and 25 mg/kg better than 15 mg/kg. The occurrence of ocular toxicity was also dose-related; >40% of adults developed toxicity at doses of >50 mg/ kg, and 0-3% at a dose of 15 mg/kg/daily. Peak serum EMB concentrations increase in relation to dose, but are significantly lower in children receiving the same dosage. In only 2 of 3811 children (0.05%) receiving EMB doses of 15-30 mg/kg was EMB stopped due to possible ocular toxicity; children of all ages can be given EMB in daily doses of 20 mg/kg (range 15-25 mg/kg) and three times weekly intermittent doses of 30 mg/kg body weight without undue concern.


Transport of the antiviral nucleoside analogs 3'-azido-3'-deoxythymidine and 2',3'-dideoxycytidine by a recombinant nucleoside transporter (rCNT) expressed in Xenopus laevis oocytes.
Expression screening in Xenopus oocytes has been used to isolate a cDNA from rat jejunal epithelium encoding an intestinal/kidney Na(+)-dependent nucleoside transporter protein named rCNT1 [J. Biol. Chem. 269:17757-17760 (1994)]. rCNT1 is predicted to have 648 amino acid residues (relative molecular mass, 71,000) with 14 potential transmembrane domains and belongs to a new family of transporter proteins. Recombinant rCNT1 transports physiological pyrimidine nucleosides and adenosine. In the current investigation, functional expression in Xenopus oocytes was used to determine whether recombinant rCNT1 also transports antiviral pyrimidine nucleoside analogs. The recombinant protein mediated Na(+)-dependent transport of both 3'-azido-3'-deoxythymidine (AZT) and 2',3'-dideoxycytidine (ddC). Apparent K(m) values of '0.5 mM were obtained for both [3H]AZT and [3H]ddC influx compared with 37 microM for [3H]uridine influx, with Vmax/Km ratios of 0.048, 0.039, and 0.57 for AZT, ddC, and uridine, respectively. Extracellular AZT and ddC stimulated rCNT1-mediated efflux of [3H]uridine from preloaded oocytes. These experiments provide direct evidence for Na(+)-dependent transport of AZT and ddC and suggest that members of the cNT family may be involved in the intestinal absorption and renal handling of pyrimidine nucleoside analogs used to treat acquired immune deficiency syndrome.


GeneiASE: Detection of condition-dependent and static allele-specific expression from RNA-seq data without haplotype information.
Allele-specific expression (ASE) is the imbalance in transcription between maternal and paternal alleles at a locus and can be probed in single individuals using massively parallel DNA sequencing technology. Assessing ASE within a single sample provides a static picture of the ASE, but the magnitude of ASE for a given transcript may vary between different biological conditions in an individual. Such condition-dependent ASE could indicate a genetic variation with a functional role in the phenotypic difference. We investigated ASE through RNA-sequencing of primary white blood cells from eight human individuals before and after the controlled induction of an inflammatory response, and detected condition-dependent and static ASE at 211 and 13021 variants, respectively. We developed a method, GeneiASE, to detect genes exhibiting static or condition-dependent ASE in single individuals. GeneiASE performed consistently over a range of read depths and ASE effect sizes, and did not require phasing of variants to estimate haplotypes. We observed condition-dependent ASE related to the inflammatory response in 19 genes, and static ASE in 1389 genes. Allele-specific expression was confirmed by validation of variants through real-time quantitative RT-PCR, with RNA-seq and RT-PCR ASE effect-size correlations r = 0.67 and r = 0.94 for static and condition-dependent ASE, respectively.


Not Just Mortality and Morbidity but Also Function: Opportunities and Challenges for Occupational Therapy in the World Health Organization's Rehabilitation 2030 Initiative.
The World Health Organization's (WHO's) new focus on rehabilitation offers great opportunities for occupational therapy. Recognizing that the problems in daily function created by the aging of the world's population and the increasing prevalence of chronic health conditions require rehabilitation to be a vital part of health systems, the World Health Organization brought a variety of stakeholders together in 2017 and 2019 to help them develop the Rehabilitation 2030 initiative. Occupational therapy was represented by the World Federation of Occupational Therapists, the American Journal of Occupational Therapy, the Canadian Journal of Occupational Therapy, editorial board members of several occupational therapy journals, and other occupational therapy researchers. A major challenge to meeting the goal of sufficient rehabilitation services to meet global need is the worldwide shortage of occupational therapists. This shortage can be attributed to a paucity of knowledge about occupational therapy as a health care profession, lack of understanding of the occupational therapy scope of practice, the cost of education required to become an occupational therapist, and the need for occupational therapy salaries to offset educational costs. Nonetheless, ongoing success in research and clinical outcomes will increase knowledge of the value of occupational therapy and increase the size of the profession. WHO's focus on function is to be lauded. Yet, occupational therapists must not be content with an emphasis on function. They also need to advocate for contextual changes that eliminate barriers to participation and engagement experienced by people with disability.


Safety and efficacy of subretinal readministration of a viral vector in large animals to treat congenital blindness.
Leber's congenital amaurosis (LCA) is a group of severe inherited retinal degenerations that are symptomatic in infancy and lead to total blindness in adulthood. Recent clinical trials using recombinant adeno-associated virus serotype 2 (rAAV2) successfully reversed blindness in patients with LCA caused by RPE65 mutations after one subretinal injection. However, it was unclear whether treatment of the second eye in the same manner would be safe and efficacious, given the potential for a complicating immune response after the first injection. Here, we evaluated the immunological and functional consequences of readministration of rAAV2-hRPE65v2 to the contralateral eye using large animal models. Neither RPE65-mutant (affected; RPE65(-/-)) nor unaffected animals developed antibodies against the transgene product, but all developed neutralizing antibodies against the AAV2 capsid in sera and intraocular fluid after subretinal injection. Cell-mediated immune responses were benign, with only 1 of 10 animals in the study developing a persistent T cell immune response to AAV2, a response that was mediated by CD4(+) T cells. Sequential bilateral injection caused minimal inflammation and improved visual function in affected animals. Thus, subretinal readministration of rAAV2 in animals is safe and effective, even in the setting of preexisting immunity to the vector, a parameter that has been used to exclude patients from gene therapy trials.


[Schistosomiasis mansoni in Bahia, Brazil: historical trends and control measures].
In order to aid the development of new approaches to schistosomiasis control, changes were analyzed in prevalence at the county level in the State of Bahia from the 1950s to the 1990s, as were determinants and the effect of community-based chemotherapy. In general, no substantial changes were observed in the basic pattern of spatial distribution of the prevalence of infection. However, during the period studied, there was an overall reduction in prevalence from 15.6% to 9.5% and an increase in prevalence rates in some counties from the western, southeastern, and northern areas of Bahia, indicative of new transmission areas. The effect of mass chemotherapy was analyzed. In the Paraguaçu Basin, where this control measure was used on a large scale, there was a reduction in prevalence similar to areas where this measure was not used. Correlation and regression analysis also failed to show links between mass chemotherapy and long term reduction in prevalence. The most powerful variables to explain these changes were those related with population dynamics. These findings strongly suggest that the reduction in prevalence observed in some areas of the State must be attributed to factors related with the spatial organization of this territory, causing a general decrease in the transmission rate, secondary to mass chemotherapy. At the same time, the incomplete form and the spatial inequalities that characterized the urbanization process created favorable conditions for the spread of schistosomiasis mansoni and the establishment of new foci.


The influence of the permeant ions thallous and potassium on inward rectification in frog skeletal muscle.
A three-electrode voltage-clamp method was used to investigate the inactivation of Tl currents through the inward rectifier of frog sartorius muscle fibres, and the interaction between the permeant ions Tl+ and K+. In 80 mM-Tl Ringer inward currents inactivated on hyperpolarization along an exponential time course, with time constants that initially increased and then fell with increasing hyperpolarization. Because of the inactivation process steady-state conductances were smaller than instantaneous conductances at all potentials in Tl Ringer. The steady-state conductance increased to a maximum value at around - 100 mV in 80 mM-Tl Ringer, and then fell with increasing hyperpolarization. In K Ringer the steady-state conductance was greater at all potentials than the instantaneous conductance because K currents activate (rather than inactivate) on hyperpolarization. Time constants of Tl inactivation were the same when measured from the decay of current during a single pulse, or from the rate of recovery from inactivation using either a two- or a three-pulse method, indicating that inactivation obeys first-order kinetics. In 80 mM-Tl Ringer steady-state inactivation increased with increasing hyperpolarization, e-fold every 48 mV. This would be consistent with the site at which inactivation occurs experiencing 0.5 of the membrane voltage field. Tl+ was more permeant than K+ through the inward rectifier, the permeability ratio PTl+/PK+ being 1.66. In solutions containing both Tl+ and K+ the membrane showed an anomalous mole-fraction dependence of conductance, the resting potential being more negative, and both instantaneous and steady-state conductances smaller than those recorded in solutions containing only Tl+ or only K+. The reduction in the amplitude of the instantaneous conductance in Tl-K mixtures was voltage-dependent, the block being initially increased and then falling with increasing hyperpolarization. Inward currents also inactivated on hyperpolarization in Tl-K mixtures. The time constants of inactivation, and the extent of inactivation which occurred, became less dependent on membrane potential in these solutions. When K+ is the major permeant ion in solution, Tl+ has a blocking effect on the currents carried by K+, and the degree of block is voltage-dependent. Increasing [Tl]o increased the block at all potentials. The results of our experiments in solutions containing both Tl+ and K+ are discussed in terms of an interaction between these ions within the channel.


Symbionts of the gut flagellate Staurojoenina sp. from Neotermes cubanus represent a novel, termite-associated lineage of Bacteroidales: description of 'Candidatus Vestibaculum illigatum'.
The symbioses between cellulose-degrading flagellates and bacteria are one of the most fascinating phenomena in the complex micro-ecosystem found in the hindgut of lower termites. However, little is known about the identity of the symbionts. One example is the epibiotic bacteria colonizing the surface of hypermastigote protists of the genus Staurojoenina. By using scanning electron microscopy, it was shown that the whole surface of Staurojoenina sp. from the termite Neotermes cubanus is densely covered with long rod-shaped bacteria of uniform size and morphology. PCR amplification of 16S rRNA genes from isolated protozoa and subsequent cloning yielded a uniform collection of clones with virtually identical sequences. Phylogenetic analysis placed them as a new lineage among the Bacteroidales, only distantly related to other uncultivated bacteria in the hindgut of other termites, including an epibiont of the flagellate Mixotricha paradoxa. The closest cultivated relative was Tannerella forsythensis (<85 % sequence identity). Fluorescence in situ hybridization with a newly designed clone-specific oligonucleotide probe confirmed that these sequences belong to the rod-shaped epibionts of Staurojoenina sp. Transmission electron microscopy confirmed the presence of a Gram-negative cell wall and revealed special attachment sites for the symbionts on the cell envelope of the flagellate host. Based on the isolated phylogenetic position and the specific association with the surface of Staurojoenina sp., we propose to classify this new taxon of Bacteroidales under the provisional name 'Candidatus Vestibaculum illigatum'.


[Is amalgam a health hazard?].
Since more than 100 years amalgam is successfully used for the functional restoration of decayed teeth. During the early 1990s the use of amalgam has been discredited by a not very objective discussion about small amounts of quicksilver that can evaporate from the material. Recent studies and reviews, however, found little to no correlation between systemic or local diseases and amalgam restorations in man. Allergic reactions are extremely rare. Most quicksilver evaporates during placement and removal of amalgam restorations. Hence it is not recommended to make extensive rehabilitations with amalgam in pregnant or nursing women. To date, there is no dental material, which can fully substitute amalgam as a restorative material. According to present scientific evidence the use of amalgam is not a health hazard.


Behavior of phosphorous and contaminants from molecular doping combined with a conventional spike annealing method.
The fabrication of future nanoscale semiconductor devices calls for precise placement of dopant atoms into their crystal lattice. Monolayer doping combined with a conventional spike annealing method provides a bottom-up approach potentially viable for large scale production. While the diffusion of the dopant was demonstrated at the start of the method, more sophisticated techniques are required in order to understand the diffusion, at the near surface, of P and contaminants such as C and O carried by the precursor, not readily accessible to direct time-of-flight secondary ion mass spectrometry measurements. By employing atom probe tomography, we report on the behavior of dopant and contaminants introduced by the molecular monolayer doping method into the first nanometers. The unwanted diffusion of C and O-related molecules is revealed and it is shown that for C and O it is limited to the first monolayers, where Si-C bonding formation is also observed, irrespective of the spike annealing temperature. From the perspective of large scale employment, our results suggest the benefits of adding a further process to the monolayer doping combined with spike annealing method, which consists of removing a sacrificial Si layer to eliminate contaminants.


Atrial fibrillation and cycling: six year follow-up of the Taupo bicycle study.
Atrial Fibrillation (AF) is the most common sustained cardiac arrhythmia, and the incidence of AF is increased markedly among elite athletes. It is not clear how lesser levels of physical activity in the general population influence AF. We asked whether participation in the Taupo Cycle Challenge was associated with increased hospital admissions due to AF, and within the cohort, whether admissions for AF were related to frequency and intensity of cycling. Participants in the 2006 Lake Taupo Cycle Challenge, New Zealand's largest mass cycling event, were invited to complete an on-line questionnaire. Those who agreed (n = 2590, response rate = 43.1%) were followed up by record linkage via the National Minimum Health Database from December 1 2006 until June 30 2013, to identify admissions to hospital due to AF. The age and gender standardized admission rate for AF was similar in the Taupo cohort (19.60 per 10,000 per year) and the national population over the same period (2006-2011) (19.45 per 10,000 per year). Within the study cohort (men only), for every additional hour spent cycling per week the risk changed by 0.90 (95% confidence interval 0.79 - 1.01). This result did not change appreciably after adjustment for age and height. Hospital admission due to AF was not increased above the national rate in this group of non-elite cyclists, and within the group the rate of AF did not increase with amount of cycling. The level of activity undertaken by this cohort of cyclists was, on average, not sufficient to increase the risk of hospitalization for AF.


Role of urokinase (uPA) and its receptor (uPAR) in invasion and metastasis of hormone-dependent malignancies.
Despite our recent advances in characterizing the molecular basis of breast and prostate cancer and their early detection with the aid of new imaging and diagnostic techniques, these cancers continue to be the leading causes of cancer-related deaths. This limited success in achieving our ultimate goal of cancer control is due to our inability to block the production of various factors produced in the later stages of these cancers that cause this high rate of mortality. A key requirement in the complex process of tumor invasion is the ability of tumor cells to produce and recruit growth factors and proteolytic enzymes within the tumor cell environment to promote neovascularization, tumor growth and promote extracellular matrix (ECM) degradation to facilitate tumor metastasis. One such protease, urokinase (uPA), has been strongly implicated in the progression of several malignancies including breast and prostate cancer. Along with uPA, its cell surface receptor (uPAR) is also believed to be involved due to its ability to recruit uPA within the tumor cell environment. In recent years, novel in vivo models of breast and prostate cancer have been developed which have clearly demonstrated the significance of uPA and uPAR in the invasion and metastases of these hormone-dependent cancers. The availability of these in vivo models has now permitted us to evaluate the molecular, chemical and immunotherapeutic strategies targeted against the uPA/uPAR system. This review describes the mechanism of uPA actions in tumor progression and analyses the usefulness of these in vivo models to authenticate uPA/uPAR as a therapeutic target and evaluates the benefits of blocking uPA/uPAR interactions alone or in combination with currently available treatment modalities against this cancer. Based on these results, there is an urgent need to develop and optimize strategies which will ultimately allow us to control the progression of these malignancies and enhance our ability to effectively manage these patients.


Bioartificial reconstruction of peripheral nerves using the rat median nerve model.
Different bioartificial tubes were recommended for peripheral nerve reconstruction in the past. In order to replace autologous nerve grafts this materials are still under review in different animal studies. Most of them are dealing with the rodent peripheral nerves. One very popular animal model to study different materials is the rat median nerve model. With its easy excess, simple behavioral tests and reliable long term results it is attractive to many scientists in this field. This review gives an overview about the past, current and future options in this model for bioartificial nerve tubes. It summarizes the evolution of successful implantation of different materials across short nerve gaps and demonstrates the obstacles arising from long nerve gaps and the problems associated to them.


Tunable infrared laser detection of pyrolysis products of explosives in soils.
A research program involving two applications of tunable infrared laser differential absorption spectroscopy (TILDAS) with multipass, long-path absorption cells to the detection of explosives contamination in soils is reported. In the first application, sensitive, specific real-time species concentration measurements by TILDAS have led to new understanding of the processes involved in explosives detection by the heating of contaminated soils and the quantification of the resulting pyrolysis gases. In the second, we present results of our calculations of the properties of astigmatic off-axis resonator absorption cells, which show that useful TILDAS path lengths can be achieved inside a cone penetrometer probe.


Exposure of Developing Brain to General Anesthesia: What Is the Animal Evidence?
Recently, the U.S. Food and Drug Administration issued an official warning to all practicing physicians regarding potentially detrimental behavioral and cognitive sequelae of an early exposure to general anesthesia during in utero and in early postnatal life. The U.S. Food and Drug Administration concern is focused on children younger than three years of age who are exposed to clinically used general anesthetics and sedatives for three hours or longer. Although human evidence is limited and controversial, a large body of scientific evidence gathered from several mammalian species demonstrates that there is a potential foundation for concern. Considering this new development in public awareness, this review focuses on nonhuman primates because their brain development is the closest to humans in terms of not only timing and duration, but in terms of complexity as well. The review compares those primate findings to previously published work done with rodents.


Coordinated Action of Aquaporins Regulates Sperm Motility in a Marine Teleost.
In marine teleosts, such as the gilthead seabream, several aquaporin paralogs are known to be expressed during the hyperosmotic induction of spermatozoon motility in seawater. Here, we used immunological inhibition of channel function to investigate the physiological roles of Aqp1aa, Aqp1ab, and Aqp7 during seabream sperm activation. Double immunofluorescence microscopy of SW-activated sperm showed that Aqp1aa and Aqp7 were respectively distributed along the flagellum and the head, whereas Aqp1ab accumulated in the head and in discrete areas toward the anterior tail. Inhibition of Aqp1aa reduced the rise of intracellular Ca(2+), which is independent of external Ca(2+) and normally occurs upon activation, and strongly inhibited sperm motility. Impaired Aqp1aa function also prevented the intracellular trafficking of Aqp8b to the mitochondrion, where it acts as a peroxiporin allowing H2O2 efflux and ATP production during activation. However, restoring the Ca(2+) levels with a Ca(2+) ionophore in spermatozoa with immunosuppressed Aqp1aa function fully rescued mitochondrial Aqp8b accumulation and sperm motility. In contrast, exposure of sperm to Aqp1ab and Aqp7 antibodies did not affect motility during the initial phase of activation, but latently compromised the trajectory and the pattern of movement. These data reveal the coordinated action of spatially segregated aquaporins during sperm motility activation in a marine teleost, where flagellar-localized Aqp1aa plays a dual Ca(2+)-dependent role controlling the initiation of sperm motility and the activation of mitochondrial detoxification mechanisms, while Aqp1ab and Aqp7 in the head and anterior tail direct the motion pattern.


First Report of Neofusicoccum australe and N. luteum Associated with Canker and Dieback of Quercus robur in Portugal.
Quercus robur L., commonly known as "pedunculate oak," is a long-lived deciduous tree native to most of Europe. It is of great ecological and forestry importance. It is also commonly cultivated as an ornamental tree in parks and gardens. Since 2009 and most likely related to increased drought periods, diseased ornamental trees have been observed in the campus of the University of Aveiro, Portugal. More than 50% of the trees are already damaged by the disease. The symptoms included twig and branch dieback and sunken necrotic bark lesions that could progress to the trunk, resulting in the death of large sections of the tree. Ascomata and conidiomata typical of Botryosphaeriaceae were observed on branches of symptomatic trees. Ascospores were hyaline, aseptate, ovoid to fusoid, and conidia were hyaline, aseptate, smooth, thin-walled, and fusiform with base truncate. Single spore isolates were obtained from samples. In culture, single ascospore isolates produced conidia similar to the ones found on the host. Diseased branch tissues were surface sterilized with 5% NaOCl, plated on potato dextrose agar (PDA), and incubated at 25°C. Fungal isolates recovered produced white aerial mycelium that darkened with age becoming grey to dark grey and conidia that were similar in all aspects to the ones produced by single spore isolates. All isolates produced on PDA a yellow pigment that diffused into the agar and disappeared after 6 to 7 days. Morphological and cultural aspects of the isolates were similar to the species Neofusicoccum luteum (Pennycook & Samuels) Crous, Slippers & A.J.L. Phillips and N. australe (Slippers, Crous & M.J. Wingf.) Crous, Slippers & A.J.L. Phillips (2). Enterobacterial Repetitive Intergenic Consensus (ERIC) PCR fingerprinting divided the isolates into two groups (1). Two isolates (CAA352 and CAA392), one from each group, were selected for further identification by sequencing of the internal transcribed spacer (ITS) region of the rDNA and part of the elongation factor 1-alpha gene (EF1-α) (Accession No. ITS: JX975212 JX975213; EF1-α: JX975210 JX975211). BLAST analysis showed that ITS and EF1-α sequences from group 1 and 2 had 99 to 100% similarity to reference cultures (including ex-type) of N. luteum and N. australe, respectively. To confirm pathogenicity and fulfill Koch's postulates, six 2-year-old seedlings of Q. robur were artificially infected with isolates CAA352 and CAA392 and kept at approximately 20 to 25°C. A shallow wound was done with a scalpel on the basal part of the stem of each seedling, a bark portion was removed aseptically and a PDA disc (0.5 cm) of an actively growing culture was placed on the wound. Control seedlings received sterile PDA discs. The inoculation site was wrapped in Parafilm to prevent desiccation. Within 8 weeks, infected seedlings developed canker lesions associated with vascular necrosis around the inoculation point. A third of the seedlings died and developed abundant pycnidia on the stem. Control seedlings remained symptomless. Both pathogens were successfully reisolated from the infected tissue. N. luteum and N. australe are increasingly reported as causing diseases to a wide range of woody hosts of economic and forestry importance (3). To our knowledge, this is the first report of both species causing dieback and canker disease on Q. robur. References: (1) A. Alves et al. Res. Microbiol. 158:112, 2007. (2) P. W. Crous et al. Stud. Mycol. 55:235, 2006. (3) B. Slippers et al. Fungal Biol. Rev. 21:90, 2007.


Metformin-like effect of Salvia officinalis (common sage): is it useful in diabetes prevention?
Common sage (Salvia officinalis L.) is among the plants that are claimed to be beneficial to diabetic patients, and previous studies have suggested that some of its extracts have hypoglycaemic effects in normal and diabetic animals. In the present study, we aimed to verify the antidiabetic effects of an infusion (tea) of common sage, which is the most common form of this plant consumed. Replacing water with sage tea for 14 d lowered the fasting plasma glucose level in normal mice but had no effect on glucose clearance in response to an intraperitoneal glucose tolerance test. This indicated effects on gluconeogenesis at the level of the liver. Primary cultures of hepatocytes from healthy, sage-tea-drinking rats showed, after stimulation, a high glucose uptake capacity and decreased gluconeogenesis in response to glucagon. Essential oil from sage further increased hepatocyte sensitivity to insulin and inhibited gluconeogenesis. Overall, these effects resemble those of the pharmaceutical drug metformin, a known inhibitor of gluconeogenesis used in the treatment and prevention of type 2 diabetes mellitus. In primary cultures of rat hepatocytes isolated from streptozotocin (STZ)-induced diabetic rats, none of these activities was observed. The present results seem to indicate that sage tea does not possess antidiabetic effects at this level. However, its effects on fasting glucose levels in normal animals and its metformin-like effects on rat hepatocytes suggest that sage may be useful as a food supplement in the prevention of type 2 diabetes mellitus by lowering the plasma glucose of individuals at risk.


Individual patient data meta-analysis of self-monitoring of blood pressure (BP-SMART): a protocol.
Self-monitoring of blood pressure is effective in reducing blood pressure in hypertension. However previous meta-analyses have shown a considerable amount of heterogeneity between studies, only part of which can be accounted for by meta-regression. This may be due to differences in design, recruited populations, intervention components or results among patient subgroups. To further investigate these differences, an individual patient data (IPD) meta-analysis of self-monitoring of blood pressure will be performed. We will identify randomised trials that have compared patients with hypertension who are self-monitoring blood pressure with those who are not and invite trialists to provide IPD including clinic and/or ambulatory systolic and diastolic blood pressure at baseline and all follow-up points where both intervention and control groups were measured. Other data requested will include measurement methodology, length of follow-up, cointerventions, baseline demographic (age, gender) and psychosocial factors (deprivation, quality of life), setting, intensity of self-monitoring, self-monitored blood pressure, comorbidities, lifestyle factors (weight, smoking) and presence or not of antihypertensive treatment. Data on all available patients will be included in order to take an intention-to-treat approach. A two-stage procedure for IPD meta-analysis, stratified by trial and taking into account age, sex, diabetes and baseline systolic BP will be used. Exploratory subgroup analyses will further investigate non-linear relationships between the prespecified variables. Sensitivity analyses will assess the impact of trials which have and have not provided IPD. This study does not include identifiable data. Results will be disseminated in a peer-reviewed publication and by international conference presentations. IPD analysis should help the understanding of which self-monitoring interventions for which patient groups are most effective in the control of blood pressure.


High levels of physical inactivity and sedentary behaviors among US immigrant children and adolescents.
To examine the prevalence and correlates of physical inactivity and sedentary behavior among immigrant and US-born children. Cross-sectional analysis using data from the 2003 National Survey of Children's Health, a telephone survey conducted between January 29, 2003, and July 1, 2004. United States. Multivariate logistic and least squares regression models were used to analyze immigrant differentials among 68 288 children aged 6 through 17 years. Main Exposure Ethnic-immigrant status. Prevalence and odds of regular physical activity, inactivity, television watching, and lack of sports participation. Physical inactivity and sedentary behaviors varied widely among children in various ethnic-immigrant groups. For example, 22.5% of immigrant Hispanic children were physically inactive compared with 9.5% of US-born white children with US-born parents. Approximately 67% of immigrant Hispanic children did not participate in sports compared with 30.2% of native Asian children. Overall, immigrant children were significantly more likely to be physically inactive and less likely to participate in sports than native children; they were, however, less likely to watch television 3 or more hours per day than native children, although the nativity gap narrowed with increasing acculturation levels. Compared with native white children, the adjusted odds of physical inactivity and lack of sports participation were both 2 times higher for immigrant Hispanic children with foreign-born parents, and the odds of television watching were 1.5 and 2.3 times higher for native Hispanic and black children, respectively. Immigrant children in each ethnic minority group generally had higher physical inactivity and lower sports participation levels than native children. To reduce disparities, health education programs need to promote physical activity among children in immigrant families.


Efficient method for establishing F1 progeny from wild populations of Anopheles mosquitoes.
The changing malaria situation in Madagascar requires additional knowledge on the physiology and behaviour of local mosquito vectors. However, the absence of established colonies for several anopheline species present in Madagascar constitutes a limiting factor. To avoid labour intensive work and uncertainty for success of establishing Anopheles colonies from Malagasy species, field collections of blood-fed females and in-tube forced oviposition were combined to reliably produce large numbers of F1 progeny. Blood-fed females were captured in zebu stables or open zebu parks. Oviposition was induced by enclosing gravid females in eppendorf tubes as initially described for Anopheles funestus. The effect of cold anaesthesia on inducing in-tube forced oviposition and on egg yield was assessed for five Anopheles species, namely Anopheles coustani, An. funestus, Anopheles mascarensis, Anopheles arabiensis and Anopheles squamosus. The production of eggs from in-tube forced oviposition and standard egg laying in cages was compared. For the five anopheline species studied, the in-tube forced oviposition method had different efficacy ranging from 35.6 to 71.1% females willing to lay eggs in tubes. Interestingly, prior anaesthesia increased significantly the proportion of ovipositing females for An. mascarensis. Prior anaesthesia has a marginal effect on the number of eggs produced. However, the overall yield in eggs collected using the in-tube forced oviposition method largely exceeds the number of eggs that can be produced by females free to oviposit in cages. The efficiency of the method allowed the production of F1 progeny in numbers sufficiently large for developing detailed analyses of the five species tested, including behavioural studies, insecticide resistance assessment and molecular characterization, as well as vector competence studies. It should be applicable to other anopheline species difficult to colonize.


Taxonomic examination of longgu (Fossilia Ossis Mastodi, "dragon bone") and a related crude drug, longchi (Dens Draconis, "dragon tooth"), from Japanese and Chinese crude drug markets.
Longgu ("dragon bone," Ryu-kotsu, Fossilia Ossis Mastodi, or Os Draconis) is the only fossil crude drug listed in the Japanese Pharmacopoeia. All longgu in the current Japanese market is imported from China, where its resources are being depleted. Therefore, effective countermeasures are urgently needed to prevent resource depletion. One possible solution is the development of a substitute made from bones of contemporary animals that are closely related to the original animal source of the current longgu. However, no research has been conducted on the original animal source of longgu, except for a report on the longgu specimens present in the Shosoin Repository. Taxonomic examination was performed on the fossil specimens related to longgu which are owned by the Museum of Osaka University, Japan. In total, 20,939 fossil fragments were examined, of which 20,886 were mammalian fossils, and 246 of these fossils were classified into nine families. The longgu specimens from the Japanese market belonged to a relatively smaller variety of taxa than those from the Chinese market. Despite the variety of taxa in longgu, medical doctors using Kampo preparations with longgu have not reported any problems due to the presence of impurities in the original animal source. These results suggest that the effect of longgu is independent of its origin as long as it is closely related to the origin of the current longgu. Thus, despite the considerable effects of fossilization, our results could help in developing an optimal substitute for longgu.


Platinum nanoparticles in nanobiomedicine.
Oxidative stress-dependent inflammatory diseases represent a major concern for the population's health worldwide. Biocompatible nanomaterials with enzymatic properties could play a crucial role in the treatment of such pathologies. In this respect, platinum nanoparticles (PtNPs) are promising candidates, showing remarkable catalytic activity, able to reduce the intracellular reactive oxygen species (ROS) levels and impair the downstream pathways leading to inflammation. This review reports a critical overview of the growing evidence revealing the anti-inflammatory ability of PtNPs and their potential applications in nanomedicine. It provides a detailed description of the wide variety of synthetic methods recently developed, with particular attention to the aspects influencing biocompatibility. Special attention has been paid to the studies describing the toxicological profile of PtNPs with an attempt to draw critical conclusions. The emerging picture suggests that the material per se is not causing cytotoxicity, while other physicochemical features related to the synthesis and surface functionalization may play a crucial role in determining the observed impairment of cellular functions. The enzymatic activity of PtNPs is also summarized, analyzing their action against ROS produced by pathological conditions within the cells. In particular, we extensively discuss the potential of these properties in nanomedicine to down-regulate inflammatory pathways or to be employed as diagnostic tools with colorimetric readout. A brief overview of other biomedical applications of nanoplatinum is also presented.


Epidemiology of stuttering in the community across the entire life span.
A randomized and stratified investigation was conducted into the epidemiology of stuttering in the community across the entire life span. Persons from households in the state of New South Wales, Australia, were asked to participate in a telephone interview. Consenting persons were given a brief introduction to the research, and details were requested concerning the number and age of the persons living in the household at the time of the interview. Interviewees were then given a description of stuttering. Based on this description, they were asked if any person living in their household stuttered (prevalence). If they answered "yes," a number of corroborative questions were asked, and permission was requested to tape over the telephone the speech of the person who stutters. Confirmation of stuttering was based on (a) a positive detection of stuttering from the tape and (b) an affirmative answer to at least one of the corroborative questions supporting the diagnosis. Results showed that the prevalence of stuttering over the whole population was 0.72%, with higher prevalence rates in younger children (1.4-1.44) and lowest rates in adolescence (0.53). Male-to-female ratios ranged from 2.3:1 in younger children to 4:1 in adolescence, with a ratio of 2.3:1 across all ages. The household member being interviewed was also asked whether anyone in the household had ever stuttered. If the answer was "yes," the same corroborative questions were asked. These data, along with the prevalence data, provided an estimate of the incidence or risk of stuttering, which was found to range from 2.1% in adults (21-50 years) to 2.8% in younger children (2-5 years) and 3.4% in older children (6-10 years). Implications of these results are discussed.


Characterization and application of surface-molecular-imprinted-polymer modified TiO2 nanotubes for removal of perfluorinated chemicals.
The removal of perfluorinated chemicals (PFCs) during wastewater reclamation is a great concern. However, the existing advanced treatment processes are inefficient for the removal of PFCs from secondary effluents of municipal wastewater treatment plants (WWTPs) because other coexistent pollutants with less environmental significance are removed simultaneously. Therefore, research on high-selectivity, low-cost removal methods is needed. The S-MIP-TiO2 nanotube (NT) photocatalysts were fabricated, characterized and tested for removal of PFCs from wastewater for the first time. Scanning electron microscopy and Fourier transform infrared spectroscopy show that the TiO2 NTs (average diameter 60 nm) were successfully imprinted with functional groups (i.e. carboxyl). The adsorption selectivity and photocatalytic activity of the S-MIP-TiO2 NTs over perfluorooctanoic acid (PFOA) were improved compared with neat TiO2 NTs and interestingly, were higher at low PFOA concentrations (10 to 100 ng/L, as normal PFC concentrations in secondary effluents) than at high concentrations (10 to 1,000 mg/L). With S-MIP-TiO2 NTs used as photocatalysts, some representative PFCs were selectively and rapidly removed from secondary effluents of a municipal WWTP. S-MIP-TiO2 NTs exhibited excellent regeneration performance. Thus, photocatalytic treatment using is promising for effective removal of PFCs from secondary effluents of municipal WWTPs.


Regulatory models and the environment: practice, pitfalls, and prospects.
Computational models support environmental regulatory activities by providing the regulator an ability to evaluate available knowledge, assess alternative regulations, and provide a framework to assess compliance. But all models face inherent uncertainties because human and natural systems are always more complex and heterogeneous than can be captured in a model. Here, we provide a summary discussion of the activities, findings, and recommendations of the National Research Council's Committee on Regulatory Environmental Models, a committee funded by the U.S. Environmental Protection Agency to provide guidance on the use of computational models in the regulatory process. Modeling is a difficult enterprise even outside the potentially adversarial regulatory environment. The demands grow when the regulatory requirements for accountability, transparency, public accessibility, and technical rigor are added to the challenges. Moreover, models cannot be validated (declared true) but instead should be evaluated with regard to their suitability as tools to address a specific question. The committee concluded that these characteristics make evaluation of a regulatory model more complex than simply comparing measurement data with model results. The evaluation also must balance the need for a model to be accurate with the need for a model to be reproducible, transparent, and useful for the regulatory decision at hand. Meeting these needs requires model evaluation to be applied over the "life cycle" of a regulatory model with an approach that includes different forms of peer review, uncertainty analysis, and extrapolation methods than those for nonregulatory models.


A deepening understanding of animal culture suggests lessons for conservation.
A key goal of conservation is to protect biodiversity by supporting the long-term persistence of viable, natural populations of wild species. Conservation practice has long been guided by genetic, ecological and demographic indicators of risk. Emerging evidence of animal culture across diverse taxa and its role as a driver of evolutionary diversification, population structure and demographic processes may be essential for augmenting these conventional conservation approaches and decision-making. Animal culture was the focus of a ground-breaking resolution under the Convention on the Conservation of Migratory Species of Wild Animals (CMS), an international treaty operating under the UN Environment Programme. Here, we synthesize existing evidence to demonstrate how social learning and animal culture interact with processes important to conservation management. Specifically, we explore how social learning might influence population viability and be an important resource in response to anthropogenic change, and provide examples of how it can result in phenotypically distinct units with different, socially learnt behavioural strategies. While identifying culture and social learning can be challenging, indirect identification and parsimonious inferences may be informative. Finally, we identify relevant methodologies and provide a framework for viewing behavioural data through a cultural lens which might provide new insights for conservation management.


Discovery of Salmonella virulence factors translocated via outer membrane vesicles to murine macrophages.
Salmonella enterica serovar Typhimurium, an intracellular pathogen and leading cause of food-borne illness, encodes a plethora of virulence effectors. Salmonella virulence factors are translocated into host cells and manipulate host cellular activities, providing a more hospitable environment for bacterial proliferation. In this study, we report a new set of virulence factors that is translocated into the host cytoplasm via bacterial outer membrane vesicles (OMV). PagK (or PagK1), PagJ, and STM2585A (or PagK2) are small proteins composed of ∼70 amino acids and have high sequence homology to each other (>85% identity). Salmonella lacking all three homologues was attenuated for virulence in a mouse infection model, suggesting at least partial functional redundancy among the homologues. While each homologue was translocated into the macrophage cytoplasm, their translocation was independent of all three Salmonella gene-encoded type III secretion systems (T3SSs)-Salmonella pathogenicity island 1 (SPI-1) T3SS, SPI-2 T3SS, and the flagellar system. Selected methods, including direct microscopy, demonstrated that the PagK-homologous proteins were secreted through OMV, which were enriched with lipopolysaccharide (LPS) and outer membrane proteins. Vesicles produced by intracellular bacteria also contained lysosome-associated membrane protein 1 (LAMP1), suggesting the possibility of OMV convergence with host cellular components during intracellular trafficking. This study identified novel Salmonella virulence factors secreted via OMV and demonstrated that OMV can function as a vehicle to transfer virulence determinants to the cytoplasm of the infected host cell.


Comparing Sublingual and Inhaled Cannabis Therapies for Low Back Pain: An Observational Open-Label Study.
Medical cannabis is becoming an acceptable treatment modality in medicine, especially for pain relief. Concurrently, cannabis use is becoming more prevalent worldwide, a public demand-driven trend despite the lack of established scientific basis. This observational open-label study sought to investigate the effectiveness of cannabis therapy for alleviating low back pain symptoms. Two types of cannabis treatment modalities were sequentially administered to chronic low back pain patients. After an initial 1-month washout period (WO1), the first modality was cannabidiol (CBD)-rich sublingual extract treatment administered for 10 months. Following another washout period, the second modality, Δ9-tetrahydrocannabinol (THC)-rich smoked inflorescence (whole dried cannabis flowers) was administered for 12 months. Enrolled in the study were 24 patients whose advanced imaging studies (i.e. computerized tomography or magnetic resonance imaging of the lumbar spine) revealed disc herniation or spinal stenosis. Three patients dropped out of extract therapy treatment but resumed study participation to receive THC-rich smoking therapy. After a minimum of 2 years, cannabis therapy had reduced lower back pain symptoms, as assessed by Oswestry Disability Index, the SF-12 patient-reported outcome questionnaire, and the visual analogue scale. Pain reduction was not significant during the extract treatment part of the study; however, pain reduction was significant during the inhaled therapy part of the study. Our findings indicate that inhaled THC-rich therapy is more effective than CBD-rich sublingual extract therapy for treating low back pain and that cannabis therapy is safe and effective for chronic low back pain.


Identification of protein tyrosine phosphatase-like IA2 (islet cell antigen 512) as the insulin-dependent diabetes-related 37/40K autoantigen and a target of islet-cell antibodies.
The majority of patients with insulin-dependent diabetes (IDDM) have Abs to 40- and/or 37-kDa tryptic fragments (37/40K-Abs) deriving from an unidentified islet cell membrane protein distinct from glutamate decarboxylase (GAD). Recently, autoantibodies against ICA512, which has identity with the protein tyrosine phosphatase-like protein IA2, were reported. In this study we have examined whether IA2/ICA512 is the Ag specificity of 37/40K-Abs, and one of the determinants of islet cell Abs (ICA) detected by immunofluorescence. Serum from 51 of 100 new onset IDDM patients immunoprecipitated 40- and/or 37-kDa insulinoma polypeptides, and 53 immunoprecipitated in vitro translated rIA2; 49 had both 37/40K-Abs and rIA2 Abs. There were strong correlations between the levels of Abs to rIA2 and both 40 kDa (r = 0.85, p < 0.0001) and 37 kDa (r = 0.70, p < 0.0001) insulinoma polypeptides. Trypsin treatment of immunoprecipitated rIA2 yielded 40- and 37-kDa fragments, and preincubation of sera with rIA2 completely inhibited binding to the insulinoma 40- and 37-kDa polypeptides. IA2 Ab levels also correlated with ICA titer in GAD-Ab negative sera, and preincubation with rIA2 reduced ICA staining intensity in sera with ICA and IA2 Abs, but not in sera with ICA in the absence of IA2 Abs. These results provide clear evidence for the identification of IA2/ICA512 as the precursor of the islet 40- and 37-kDa polypeptide autoantigens and as one of the ICA specificities. Combined detection of Abs to IA2 and GAD65 in a single radio-binding assay identified Abs in 88 of 100 IDDM patients, and potentially facilitates population screening for IDDM risk assessment.


Community care in Europe. The Aged in Home Care project (AdHOC).
Community care for older people is increasing dramatically in most European countries as the preferred option to hospital and long-term care. While there has been a rapid expansion in Evidence-Based Medicine, apart from studies of specific interventions such as home visiting and hospital at home (specialist visits or hospital services provided to people in their own homes in the community), there is little evidence of characteristics of the recipients of community care services or the organisation of services that produce the best outcomes for them and their informal carers. The AdHOC Study was designed to compare outcomes of different models of community care using a structured comparison of services and a comprehensive standardised assessment instrument across 11 European countries. This paper describes the study and baseline data. 4,500 people 65 years and older already receiving home care services within the urban areas selected in each country were randomly sampled. They were assessed with the MDS-HC (Minimum Data Set-Home Care) instrument, containing over 300 items, including socio-demographic, physical and cognitive characteristics of patients as well as medical diagnoses and medications received. These data were linked to information on the setting, services structures and services utilization, including use of hospital and long-term care. After baseline assessment, patients were re-evaluated at 6 months with an abbreviated version of the instrument, and then at the end of one year. Data collection was performed by specially-trained personnel. In this paper, socio-demographics, physical and cognitive function and provision of hours of formal care are compared between countries at baseline. The final study sample comprised 3,785 patients; mean age was 82+/-7.2 years, 74.2% were females. Marital and living status reflected close family relationships in southern Europe relative to Nordic countries, where 5 times as many patients live alone. Recipients of community care in France and Italy are characterised by very high physical and cognitive impairment compared with those in northern Europe, who have comparatively little impairment in Activities of Daily Living and cognitive function. The provision of formal care to people with similar dependency varies extremely widely with very little formal care in Italy and more than double the average across all levels of dependency in the UK. The AdHOC study, by virtue of the use of a common comprehensive standardised assessment instrument, is a unique tool in examining older recipients of community care services in European countries and their widely varied organisation. The extreme differences seen in dependency and hours of care illustrate the probable contribution the study will make to developing an evidence based on the structure, quantity and targeting of community care, which will have major policy implications.


Influence of organic and colloidal fouling on the removal of sulphamethoxazole by nanofiltration membranes.
This study investigated the effects of organic and colloidal fouling on the removal of a representative micropollutant sulphamethoxazole by two commercially available NF membranes. Alginate, bovine serum albumin and colloidal silica were selected as model foulants to simulate hydrophilic and hydrophobic organic fractions, and colloidal matter that are often found in treated effluent and surface water. Membrane fouling was related to the membrane and foulant characteristics and subsequently the separation behaviour of the micropollutant sulphamethoxazole under different solution pH. On the basis of these results, it was confirmed that membrane fouling is strongly dependent on both the foulant and membrane characteristics. The complex relationship among retention mechanisms, fouling mechanisms and the effects of fouling on retention was systematically delineated. Of the three model foulants selected for this study, colloidal fouling resulted in the most significant reduction in retention of sulphamethoxazole as well as inorganic salts, while flux decline as a result of colloidal fouling was quite moderate. Reduction in retention caused by fouling was attributed to a phenomenon known as cake-enhance concentration polarisation, which was a predominant mechanism of colloidal fouling. In addition, the reported results suggested that the effect of fouling on retention is also membrane pore size dependent.


Calcineurin and Sorting-Related Receptor with A-Type Repeats Interact to Regulate the Renal Na⁺-K⁺-2Cl⁻ Cotransporter.
The furosemide-sensitive Na(+)-K(+)-2Cl(-)-cotransporter (NKCC2) is crucial for NaCl reabsorption in kidney thick ascending limb (TAL) and drives the urine concentrating mechanism. NKCC2 activity is modulated by N-terminal phosphorylation and dephosphorylation. Serine-threonine kinases that activate NKCC2 have been identified, but less is known about phosphatases that deactivate NKCC2. Inhibition of calcineurin phosphatase has been shown to stimulate transport in the TAL and the distal convoluted tubule. Here, we identified NKCC2 as a target of the calcineurin Aβ isoform. Short-term cyclosporine administration in mice augmented the abundance of phospho-NKCC2, and treatment of isolated TAL with cyclosporine increased the chloride affinity and transport activity of NKCC2. Because sorting-related receptor with A-type repeats (SORLA) may affect NKCC2 phosphoregulation, we used SORLA-knockout mice to test whether SORLA is involved in calcineurin-dependent modulation of NKCC2. SORLA-deficient mice showed more calcineurin Aβ in the apical region of TAL cells and less NKCC2 phosphorylation and activity compared with littermate controls. In contrast, overexpression of SORLA in cultured cells reduced the abundance of endogenous calcineurin Aβ. Cyclosporine administration rapidly normalized the abundance of phospho-NKCC2 in SORLA-deficient mice, and a functional interaction between calcineurin Aβ and SORLA was further corroborated by binding assays in rat kidney extracts. In summary, we have shown that calcineurin Aβ and SORLA are key components in the phosphoregulation of NKCC2. These results may have clinical implications for immunosuppressive therapy using calcineurin inhibitors.


Establishment of Coral-Bacteria Symbioses Reveal Changes in the Core Bacterial Community With Host Ontogeny.
Bacterial communities are fundamental symbionts of corals. However, the process by which bacterial communities are acquired across the life history of corals, particularly in larval and early juvenile stages, is still poorly characterized. Here, transfer of bacteria of the Scleractinian coral Acropora digitifera from adults to spawned egg-sperm bundles was analyzed, as well as acquisition across early developmental stages (larvae and newly settled spat), and 6-month-old juveniles. Larvae were reared under manipulated environmental conditions to determine the source (maternal, seawater, or sediment) of bacteria likely to establish symbiotic relationships with the host using amplicon sequencing of the 16S rRNA gene. Maternal colonies directly transferred bacteria from the families Rhodobacteraceae, Cryomorphaceae, and Endozoicimonaceae to egg-sperm bundles. Furthermore, significant differences in the microbial community structure were identified across generations, yet the structure of the coral bacterial community across early life history stages was not impacted by different environmental rearing conditions. These data indicate that the uptake and structure of bacterial communities is developmentally, rather than environmentally, regulated. Both maternal coral colonies and ubiquitous bacteria found across environmental substrates represent a potential source of symbionts important in establishing the coral microbiome. Uniquely, we report the presence of variation with ontogeny of both the core and resident bacterial communities, supporting the hypothesis that microbial communities are likely to play specific roles within the distinct life history stages of the coral host.


Implementation of 3D Printing Technology in the Field of Prosthetics: Past, Present, and Future.
There is an interesting and long history of prostheses designed for those with upper-limb difference, and yet issues still persist that have not yet been solved. Prosthesis needs for children are particularly complex, due in part to their growth rates. Access to a device can have a significant impact on a child's psychosocial development. Often, devices supporting both cosmetic form and user function are not accessible to children due to high costs, insurance policies, medical availability, and their perceived durability and complexity of control. These challenges have encouraged a grassroots effort globally to offer a viable solution for the millions of people living with limb difference around the world. The innovative application of 3D printing for customizable and user-specific hardware has led to open-source Do It Yourself "DIY" production of assistive devices, having an incredible impact globally for families with little recourse. This paper examines new research and development of prostheses by the maker community and nonprofit organizations, as well as a novel case study exploring the development of technology and the training methods available. These design efforts are discussed further in the context of the medical regulatory framework in the United States and highlight new associated clinical studies designed to measure the quality of life impact of such devices.


Integrating Human Factors Engineering and Information Processing Approaches to Facilitate Evaluations in Criminal Justice Technology Research.
Evaluations are routinely conducted by government agencies and research organizations to assess the effectiveness of technology in criminal justice. Interdisciplinary research methods are salient to this effort. Technology evaluations are faced with a number of challenges including (1) the need to facilitate effective communication between social science researchers, technology specialists, and practitioners, (2) the need to better understand procedural and contextual aspects of a given technology, and (3) the need to generate findings that can be readily used for decision making and policy recommendations. Process and outcome evaluations of technology can be enhanced by integrating concepts from human factors engineering and information processing. This systemic approach, which focuses on the interaction between humans, technology, and information, enables researchers to better assess how a given technology is used in practice. Examples are drawn from complex technologies currently deployed within the criminal justice system where traditional evaluations have primarily focused on outcome metrics. Although this evidence-based approach has significant value, it is vulnerable to fully account for human and structural complexities that compose technology operations. Guiding principles for technology evaluations are described for identifying and defining key study metrics, facilitating communication within an interdisciplinary research team, and for understanding the interaction between users, technology, and information. The approach posited here can also enable researchers to better assess factors that may facilitate or degrade the operational impact of the technology and answer fundamental questions concerning whether the technology works as intended, at what level, and cost.


Mercury-associated DNA hypomethylation in polar bear brains via the LUminometric Methylation Assay: a sensitive method to study epigenetics in wildlife.
In this paper we describe a novel approach that may shed light on the genomic DNA methylation of organisms with non-resolved genomes. The LUminometric Methylation Assay (LUMA) is permissive for genomic DNA methylation studies of any genome as it relies on the use of methyl-sensitive and -insensitive restriction enzymes followed by polymerase extension via Pyrosequencing technology. Here, LUMA was used to characterize genomic DNA methylation in the lower brain stem region from 47 polar bears subsistence hunted in central East Greenland between 1999 and 2001. In these samples, average genomic DNA methylation was 57.9% +/- 6.69 (SD; range was 42.0 to 72.4%). When genomic DNA methylation was related to brain mercury (Hg) exposure levels, an inverse association was seen between these two variables for the entire study population (P for trend = 0.17). After dichotomizing animals by gender and controlling for age, a negative trend was seen amongst male animals (P for trend = 0.07) but no associations were found in female bears. Such sexually dimorphic responses have been found in other toxicological studies. Our results show that genomic DNA methylation can be quantitatively studied in a highly reproducible manner in tissue samples from a wild organism with a non-resolved genome. As such, LUMA holds great promise as a novel method to explore consequential questions across the ecological sciences that may require an epigenetic understanding.


Can patients predict which consultations can be dealt with by telephone?
The use of telephone consultations to reduce the workload of general practitioners is well established both in this country and abroad. The principal aim of this study was to discover the proportion of consultations currently carried out in the surgery that would be suitable, for both doctor and patient, to be managed over the telephone. The second aim was to establish what proportion of such consultations could be predicted.


Family meals. Associations with weight and eating behaviors among mothers and fathers.
Few studies have looked at the relationship between family meals and adult weight and health behaviors. The current study investigates the association between frequency of family meals and mothers' and fathers' body mass index (BMI), dietary intake, dieting behaviors and binge eating. Data from Project F-EAT (Families and Eating and Activity in Teens) were used for the current analysis. Socio-economically and racially/ethnically diverse mothers and fathers (n=3488) of adolescents participating in a multi-level population-based study (EAT 2010) completed surveys mailed to their homes. Predicted means or probabilities were calculated for each outcome variable at each level of family meal frequency. Interactions between race/ethnicity and marital status with family meals were evaluated in all models. Overall, results indicated that having more frequent family meals was associated with increased consumption of fruits and vegetables for mothers and fathers, after adjusting for age, educational attainment, marital status and race/ethnicity. Other findings including less fast food intake for fathers and fewer dieting and binge eating behaviors for mothers were significantly associated with family meal frequency, but not consistently across all family meal categories or with BMI. Interactions by race/ethnicity and marital status were non-significant, indicating that family meals may be important for more healthful dietary intake across race and marital status. Future research should confirm findings in longitudinal analyses to identify temporality and strength of associations.


Balancing Benefits and Risks of Immortal Data: Participants' Views of Open Consent in the Personal Genome Project.
An individual's health, genetic, or environmental-exposure data, placed in an online repository, creates a valuable shared resource that can accelerate biomedical research and even open opportunities for crowd-sourcing discoveries by members of the public. But these data become "immortalized" in ways that may create lasting risk as well as benefit. Once shared on the Internet, the data are difficult or impossible to redact, and identities may be revealed by a process called data linkage, in which online data sets are matched to each other. Reidentification (re-ID), the process of associating an individual's name with data that were considered deidentified, poses risks such as insurance or employment discrimination, social stigma, and breach of the promises often made in informed-consent documents. At the same time, re-ID poses risks to researchers and indeed to the future of science, should re-ID end up undermining the trust and participation of potential research participants. The ethical challenges of online data sharing are heightened as so-called big data becomes an increasingly important research tool and driver of new research structures. Big data is shifting research to include large numbers of researchers and institutions as well as large numbers of participants providing diverse types of data, so the participants' consent relationship is no longer with a person or even a research institution. In addition, consent is further transformed because big data analysis often begins with descriptive inquiry and generation of a hypothesis, and the research questions cannot be clearly defined at the outset and may be unforeseeable over the long term. In this article, we consider how expanded data sharing poses new challenges, illustrated by genomics and the transition to new models of consent. We draw on the experiences of participants in an open data platform-the Personal Genome Project-to allow study participants to contribute their voices to inform ethical consent practices and protocol reviews for big-data research.


Flexion mobilizations with movement techniques: the immediate effects on range of movement and pain in subjects with low back pain.
This study investigates the immediate effects of flexion mobilizations with movement techniques (MWMs) on spinal range of movement in individuals with low back pain and also their impact on pain. A preliminary attempt has been made to describe the clinical profiles of subjects who were thought to benefit from MWMs. A small-scale explanatory study was conducted using a crossover design, placebo-controlled, with subjects and assessors blinded. After assessment by physiotherapists, 26 subjects with low back pain with pain on lumbar flexion, thought to be appropriate for treatment with MWMs, participated. Subjects received an MWM intervention and a placebo intervention in a randomized order. Lumbar spinal flexion and extension and pain during flexion were recorded immediately before and after each intervention, using double inclinometry and visual analogue scales. Mean spinal range of movement increased significantly with the MWM intervention, as compared with the placebo (true flexion: MWMs 49.2 degrees [SD 16.4], placebo 45.3 degrees [SD 14.1], P = .005; total flexion: MWMs 76.7 degrees [SD 22.4], placebo 69.7 degrees [SD 21.5], P = .005). Mean pain scores did not change. The MWMs produced statistically significant, but small, immediate spinal mobility increases but no pain reduction when compared with placebo. By introducing clinical judgment into the subject selection process for the trial, 19 (73%) of 26 subjects benefited from MWMs techniques in terms of range of movement and/or pain intensity, whereas 9 (35%) subjects showed such changes with the placebo intervention.


Nonadjunctive Use of Continuous Glucose Monitoring for Diabetes Treatment Decisions.
While self-monitoring of blood glucose (SMBG) is the current standard used by people with diabetes to manage glucose levels, recent improvements in accuracy of continuous glucose monitoring (CGM) technology are making it very likely that diabetes-related treatment decisions will soon be made based on CGM values alone. Nonadjunctive use of CGM will lead to a paradigm shift in how patients manage their glucose levels and will require substantial changes in how care providers educate their patients, monitor their progress, and provide feedback to help them manage their diabetes. The approval to use CGM nonadjunctively is also a critical step in the pathway toward FDA approval of an artificial pancreas system, which is further expected to transform diabetes care for people with type 1 diabetes. In this article, we discuss how nonadjunctive CGM is expected to soon replace routine SMBG and how this new usage scenario is expected to transform health outcomes and patient care.


Polarization attraction using counter-propagating waves in optical fiber at telecommunication wavelengths.
In this work, we report the experimental observation of a polarization attraction process which can occur in optical fibers at telecommunication wavelengths. More precisely, we have numerically and experimentally shown that a polarization attractor, based on the injection of two counter-propagating waves around 1.55microm into a 2-m long high nonlinear fiber, can transform any input polarization state into a unique well-defined output polarization state.


Conservation and management of the endangered Fiji sago palm, Metroxylon vitiense, in Fiji.
Recovery planning is a key component of many threatened species conservation initiatives and can be a powerful awareness raising tool. One of the largest impediments to conservation efforts in the Pacific region however, is the lack of ecological data and its subsequent effects on the development of feasible and useful recovery plans for threatened species. Without these plans, the understaffed, underfunded and often technically ill-equipped conservation agencies face huge difficulties in planning, prioritizing and conducting conservation activities to adequately protect biodiversity. The Fiji sago palm, Metroxylon vitiense, is an endemic endangered palm species whose survival is heavily dependent on a feasible species recovery plan. It is geographically restricted and threatened by habitat destruction and overexploitation for thatch for the tourism industry and palm heart consumption by local consumers. Despite its threatened status, M. vitiense is not currently protected by national or international legislation. Recent field surveys and extensive stakeholder consultation have resulted in the production of a species recovery plan highlighting the importance of the species and advocating sustainable harvesting rather than complete bans to promote conservation. This article summarizes the recovery plan and its current effects on the status of M. vitiense in Fiji. We also discuss the role of different stakeholders in the conservation of M. vitiense, including the absence of significant behavioral changes by the largest consumer - the tourism industry, and the importance of recovery plans for biodiversity conservation in the Pacific.


Attosecond imaging.
The natural timescale for electron dynamics reaches down to the attosecond domain. Following the discovery of attosecond laser pulses, about a decade ago, attosecond science has developed into a vibrant, new research field, where the motion of single or multiple electrons and, in molecules, the coupling of electronic and nuclear motion, can be investigated, on attosecond to few-femtosecond timescales. Attosecond experiments require suitable observables. This review describes how "attosecond imaging", basing itself on kinetic energy and angle-resolved detection of photoelectrons and fragment ions using a velocity map imaging (VMI) spectrometer, has been exploited in a number of pump-probe experiments. The use of a VMI spectrometer in attosecond experiments has allowed the characterization of attosecond pulse trains and isolated attosecond pulses, the elucidation of continuum electron dynamics and wave packet interferometry in atomic photoionization and the observation of electron localization in dissociative molecular photoionization.


Factors associated with referral and completion of genetic counseling in women with epithelial ovarian cancer.
The National Comprehensive Cancer Network recommends that all women diagnosed with epithelial ovarian cancer undergo genetic testing, as the diagnosis of pathogenic variants may inform cancer survival and impact treatment options. The objective of this study was to assess factors associated with referral to genetic counseling in women with epithelial ovarian cancer. A retrospective cohort study identified women with epithelial ovarian cancer from 2012 to 2017 at Massachusetts General Hospital and North Shore Medical Center, a community hospital affiliated with Massachusetts General Hospital. Multivariate logistic regression evaluated how race, age, stage, year of diagnosis, insurance status, family history of breast or ovarian cancer, and language relates to the receipt of genetic counseling. Of the total 276 women included, 73.9% were referred for genetic screening, of which 90.7% attended a genetic counseling visit. Older women were less likely to undergo genetic counseling (age ≥70 years: OR 0.26, 95% CI 0.07-0.94, p=0.04). Women who died within 365 days of initial oncology consult rarely reached a genetic counselor (OR 0.05, 95% CI 0.01-0.24, p<0.001). Women with a family history of breast or ovarian cancer were more likely to undergo counseling (OR 3.27, 95% CI 1.74-6.15, p<0.001). There was no difference in receipt of genetic counseling by race, stage, year of diagnosis, insurance status, or language. Older women with epithelial ovarian cancer and those who died within 1 year of initiation of care were less likely to undergo recommended genetic counseling. Race, insurance status, and language were not identified as predictive factors, although we were limited in this assessment by small sample size.


Fast principal component analysis of large-scale genome-wide data.
Principal component analysis (PCA) is routinely used to analyze genome-wide single-nucleotide polymorphism (SNP) data, for detecting population structure and potential outliers. However, the size of SNP datasets has increased immensely in recent years and PCA of large datasets has become a time consuming task. We have developed flashpca, a highly efficient PCA implementation based on randomized algorithms, which delivers identical accuracy in extracting the top principal components compared with existing tools, in substantially less time. We demonstrate the utility of flashpca on both HapMap3 and on a large Immunochip dataset. For the latter, flashpca performed PCA of 15,000 individuals up to 125 times faster than existing tools, with identical results, and PCA of 150,000 individuals using flashpca completed in 4 hours. The increasing size of SNP datasets will make tools such as flashpca essential as traditional approaches will not adequately scale. This approach will also help to scale other applications that leverage PCA or eigen-decomposition to substantially larger datasets.


Expansion of microsatellites on evolutionary young Y chromosome.
Sex chromosomes are an ideal system to study processes connected with suppressed recombination. We found evidence of microsatellite expansion, on the relatively young Y chromosome of the dioecious plant sorrel (Rumex acetosa, XY1Y2 system), but no such expansion on the more ancient Y chromosomes of liverwort (Marchantia polymorpha) and human. The most expanding motifs were AC and AAC, which also showed periodicity of array length, indicating the importance of beginnings and ends of arrays. Our data indicate that abundance of microsatellites in genomes depends on the inherent expansion potential of specific motifs, which could be related to their stability and ability to adopt unusual DNA conformations. We also found that the abundance of microsatellites is higher in the neighborhood of transposable elements (TEs) suggesting that microsatellites are probably targets for TE insertions. This evidence suggests that microsatellite expansion is an early event shaping the Y chromosome where this process is not opposed by recombination, while accumulation of TEs and chromosome shrinkage predominate later.


Fluid biopsy in patients with metastatic prostate, pancreatic and breast cancers.
Hematologic spread of carcinoma results in incurable metastasis; yet, the basic characteristics and travel mechanisms of cancer cells in the bloodstream are unknown. We have established a fluid phase biopsy approach that identifies circulating tumor cells (CTCs) without using surface protein-based enrichment and presents them in sufficiently high definition (HD) to satisfy diagnostic pathology image quality requirements. This 'HD-CTC' assay finds >5 HD-CTCs mL(-1) of blood in 80% of patients with metastatic prostate cancer (n = 20), in 70% of patients with metastatic breast cancer (n = 30), in 50% of patients with metastatic pancreatic cancer (n = 18), and in 0% of normal controls (n = 15). Additionally, it finds HD-CTC clusters ranging from 2 HD-CTCs to greater than 30 HD-CTCs in the majority of these cancer patients. This initial validation of an enrichment-free assay demonstrates our ability to identify significant numbers of HD-CTCs in a majority of patients with prostate, breast and pancreatic cancers.


Prevalence and clinical pattern of paediatric HIV infection at the University College Hospital, Ibadan, Nigeria: a prospective cross-sectional study.
The prevalence of Paediatric HIV infection is largely unknown in many countries in sub-Saharan Africa. This study was aimed at determining the prevalence, clinical pattern of HIV infection and outcome among new patients aged <15 years using age-specific diagnostic methods. A prospective cross sectional study was carried out using the provider initiated HIV testing and counselling (PITC) model. HIV rapid test in parallel was used for screening and confirmation was with HIV DNA PCR in children <18 months and Western Blot in children ≥ 18 months. A total of 600 children were enrolled with ages ranging between one day and 179 months. Male: female ratio was 1.2:1. HIV seroprevalence was 12.3% and after confirmatory tests, the prevalence was 10%. Fourteen (37.8%) of the children aged less 18 months were exposed but not infected. Mother-to-child transmission accounted for 93.3% of cases. Features predictive of HIV infection were diarrhoea, cough, weight loss, ear discharge generalized lymphadenopathy, presence of skin lesions, parotid swelling and oral thrush. About 75% presented in advanced or severe clinical stages of the disease, 56.8% had severe immunodeficiency while 50% had viral loads more than 100,000 copies/ml. Mortality rate was 14.3% among HIV positive compared with 11.3% in HIV negative children but was not significant. Among the HIV positive children, 26.7% were orphans. The prevalence rate of HIV infection among new patients screened using the PITC model was high, majority resulting from mother-to-child transmission. Most children presented in advanced stages of the disease and mortality rate among them was high. Though, the study site being a referral centre might have contributed to the high prevalence observed in this study, there is a need to expand access to PMTCT services, ensure implementation of PITC in paediatric settings and expand support services for HIV infected children.


Nonconventional Doses of Somatostatin Analogs in Patients With Progressing Well-Differentiated Neuroendocrine Tumor.
To evaluate the antiproliferative activity and safety of nonconventional high doses of somatostatin analogs (HD-SSA) in patients with well-differentiated gastroenteropancreatic (GEP) neuroendocrine tumors (NET) with radiological disease progression according to Response Evaluation Criteria in Solid Tumors (RECIST) criteria on a previous treatment. A retrospective analysis of prospectively maintained databases from 13 Italian NET-dedicated centers was performed. Main inclusion criteria were: well-differentiated G1 or G2 GEP-NET, progressive disease on a previous treatment, and subsequent treatment with HD-SSA (either by increased administered dose [dose intensity] or shortened interval between administrations [dose density]). Main endpoints were progression-free survival (PFS) and safety. Of 198 patients, 140 matched inclusion criteria and were included in the analysis. Overall, median PFS was 31 months. Use of HD-SSA as second-line treatment was associated with reduced risk for progression or death compared with third- or further-line treatment (HR: 2.12; P = 0.004). There was no difference in PFS between HD-SSA by increased dose density (N = 133; 95%) or intensity (N = 7; 5%). Partial response according to RECIST criteria was observed in 12 patients (8.6%), and stable disease was achieved in 106 (75.7%) patients. Adverse events occurred in 21 patients (15.0%), 2 of whom had grade 3 biliary stone disease. No patients discontinued HD-SSA treatment due to adverse events. HD-SSA is an active and safe treatment option in patients with progressive well-differentiated GEP-NET. The high rate of objective responses observed deserves prospective validation in ad hoc clinical trials.