Effects of xylene and alcohol on vestibular and visual functions in man.
Ten healthy male volunteers were exposed for 4 h to two concentrations [6 and 11.5 mumol/l (636 and 1,218 mg/m3)] of m-xylene or/and given single doses (0.4 and 0.8 g/kg) of ethyl alcohol. Exposure to two xylene concentrations combined with the higher dose (0.8 g/kg) of alcohol was also conducted. Vestibular functions (positional nystagmus with electronystagmography, body balance) and visual function (flicker fusion) were measured. Both alcohol doses increased body sway and the intensity of nystagmus more than either concentration of xylene did, but they had little effect on visual functions. The effects of alcohol on vestibular functions were dose-dependent. The effects of xylene alone on the vestibular system were rather small, and those on the visual functions negligible. The combined effect of alcohol and the lower concentration of xylene (6 mumol/l) on body sway was additive, but the higher xylene concentration (11.5 mumol/l) antagonized the effect of alcohol on body sway and positional nystagmus. Two subjects experienced nausea and vomited exposure to alcohol and the higher xylene concentration. Mild impairment in visual functions was noted in the combined exposure. Alcohol significantly increased the blood m-xylene concentrations, a finding that suggests that their antagonism was pharmacodynamic rather than pharmacokinetic.


Yield Loss in Cereals, Caused by Fusarium culmorum and F. pseudograminearum, Is Related to Fungal DNA in Soil Prior to Planting, Rainfall, and Cereal Type.
In southeastern Australia, Fusarium crown rot, caused by Fusarium culmorum or F. pseudograminearum, is an increasingly important disease of cereals. Because in-crop control options are limited, it is important for growers to know prior to planting which fields are at risk of yield loss from crown rot. Understanding the relationships between crown rot inoculum and yield loss would assist in assessing the risk of yield loss from crown rot in fields prior to planting. Thirty-five data sets from crown rot management experiments conducted in the states of South Australia and Victoria during the years 2005 to 2010 were examined. Relationships between Fusarium spp. DNA concentrations (inoculum) in soil samples taken prior to planting and disease development and grain yield were evaluated in seasons with contrasting seasonal rainfall. F. culmorum and F. pseudograminearum DNA concentrations in soil prior to planting were found to be positively related to crown rot expression (stem browning and whiteheads) and negatively related to grain yield of durum wheat, bread wheat, and barley. Losses from crown rot were greatest when rainfall during September and October (crop maturation) was below the long-term average. Losses from crown rot were greater in durum wheat than bread wheat and least in barley. Yield losses from F. pseudograminearum were similar to yield losses from F. culmorum. Yield loss patterns were consistent across experiments and between states; therefore, it is reasonable to expect that similar relationships will occur over broad geographic areas. This suggests that quantitative polymerase chain reaction technology and soil sampling could be powerful tools for assessing crown rot inoculum concentrations prior to planting and predicting the risk of yield loss from crown rot wherever this disease is an issue.


Molecular Detection of Filamentous Fungi in Formalin-Fixed Paraffin-Embedded Specimens in Invasive Fungal Wound Infections Is Feasible with High Specificity.
Trauma-related invasive fungal wound infections (IFIs) are associated with significant morbidity and mortality. Early identification and treatment are critical. Traditional identification methods (e.g., fungal cultures and histopathology) can be delayed and insensitive. We assessed a PCR-based sequencing assay for rapid identification of filamentous fungi in formalin-fixed paraffin-embedded (FFPE) specimens obtained from combat casualties injured in Afghanistan. Blinded FFPE specimens from cases (specimens positive on histopathology) and controls (specimens negative on histopathology) were submitted for evaluation with a panfungal PCR. The internal transcribed spacer 2 (ITS2) region of the fungal ribosomal repeat was amplified and sequenced. The PCR results were compared with findings from histopathology and/or culture. If injury sites contributed multiple specimens, findings for the site were collapsed to the site level. We included 64 case subjects (contributing 95 sites) and 102 controls (contributing 118 sites). Compared to histopathology, panfungal PCR was specific (99%), but not as sensitive (63%); however, sensitivity improved to 83% in specimens from sites with angioinvasion. Panfungal PCR identified fungi of the order Mucorales in 33 of 44 sites with angioinvasion (75%), whereas fungal culture was positive in 20 of 44 sites (45%). Saksenaea spp. were the dominant fungi identified by PCR in specimens from angioinvasion sites (57%). Panfungal PCR is specific, albeit with lower sensitivity, and performs better at identifying fungi of the order Mucorales than culture. DNA sequencing offers significant promise for the rapid identification of fungal infection in trauma-related injuries, leading to more timely and accurate diagnoses.


Survivorship Care Plans: Health Actions Taken and Satisfaction After Use.
To examine the use of and assess patient satisfaction with survivorship care plans (SCPs). 189 cancer survivors recruited from five cancer treatment center locations (Avera Cancer Institute in Aberdeen, Mitchell, Sioux Falls, and Yankton; Sanford Cancer Center in Sioux Falls) and one auxiliary specialty center (Urology Specialists in Sioux Falls), all in South Dakota. A written survey was completed by participants before and three months after receiving an SCP. Associations between demographics and cancer-related characteristics and use of the SCP were evaluated using chi-square tests. Logistic regression was used to determine factors associated with any use of the SCP, health actions attributable to the SCP, and satisfaction with the SCP. The most frequently reported uses of the SCP were to share with spouse or partner, inform about symptoms, and ask physician or nurse about concerns. SCP use, health actions taken, and satisfaction with the SCP were associated with gender, marital status, and main cancer type. Nurses should promote SCPs because they are valued and used by survivors for follow-up care.


Todralazine protects zebrafish from lethal effects of ionizing radiation: role of hematopoietic cell expansion.
The Johns Hopkins Clinical Compound Library (JHCCL), a collection of Food and Drug Administration (FDA)-approved small molecules (1400), was screened in silico for identification of novel β2AR blockers and tested for hematopoietic stem cell (HSC) expansion and radioprotection in zebrafish embryos. Docking studies, followed by the capacity to hasten erythropoiesis, identified todralazine (Binding energy, -8.4 kcal/mol) as a potential HSC-modulating agent. Todralazine (5 μM) significantly increased erythropoiesis in caudal hematopoietic tissue (CHT) in wild-type and anemic zebrafish embryos (2.33- and 1.44-folds, respectively) when compared with untreated and anemic control groups. Todralazine (5 μM) treatment also led to an increased number of erythroid progenitors, as revealed from the increased expression of erythroid progenitor-specific genes in the CHT region. Consistent with these effects, zebrafish embryos, Tg(cmyb:gfp), treated with 5 μM todralazine from 24 to 36 hours post fertilization (hpf) showed increased (approximately two-folds) number of HSCs at the aorta-gonad-mesonephros region (AGM). Similarly, expression of HSC marker genes, runx1 (3.3-folds), and cMyb (1.41-folds) also increased in case of todralazine-treated embryos, further supporting its HSC expansion potential. Metoprolol, a known beta blocker, also induced HSC expansion (1.36- and 1.48-fold increase in runx1 and cMyb, respectively). Todralazine (5 μM) when added 30 min before 20 Gy gamma radiation, protected zebrafish from radiation-induced organ toxicity, apoptosis, and improved survival (80% survival advantage over 6 days). The 2-deoxyribose degradation test further suggested hydroxyl (OH) radical scavenging potential of todralazine, and the same is recapitulated in vivo. These results suggest that todralazine is a potential HSC expanding agent, which might be acting along with important functions, such as antioxidant and free radical scavenging, in manifesting radioprotection.


Albendazole therapy and enteric parasites in United States-bound refugees.
Beginning on May 1, 1999, the Centers for Disease Control and Prevention (CDC) recommended presumptive treatment of refugees for intestinal parasites with a single dose of albendazole (600 mg), administered overseas before departure for the United States. We conducted a retrospective cohort study involving 26,956 African and Southeast Asian refugees who were screened by means of microscopical examination of stool specimens for intestinal parasites on resettlement in Minnesota between 1993 and 2007. Adjusted prevalence ratios for intestinal nematodes, schistosoma species, giardia, and entamoeba were calculated among refugees who migrated before versus those who migrated after the CDC recommendation of presumptive predeparture albendazole treatment. Among 4370 untreated refugees, 20.8% had at least one stool nematode, most commonly hookworm (in 9.2%). Among 22,586 albendazole-treated refugees, only 4.7% had one or more nematodes, most commonly trichuris (in 3.9%). After adjustment for sex, age, and region, albendazole-treated refugees were less likely than untreated refugees to have any nematodes (prevalence ratio, 0.19), ascaris (prevalence ratio, 0.06), hookworm (prevalence ratio, 0.07), or trichuris (prevalence ratio, 0.27) but were not less likely to have giardia or entamoeba. Schistosoma ova were identified exclusively among African refugees and were less prevalent among those treated with albendazole (prevalence ratio, 0.60). After implementation of the albendazole protocol, the most common pathogens among 17,011 African refugees were giardia (in 5.7%), trichuris (in 5.0%), and schistosoma (in 1.8%); among 5575 Southeast Asian refugees, only giardia remained highly prevalent (present in 17.2%). No serious adverse events associated with albendazole use were reported. Presumptive albendazole therapy administered overseas before departure for the United States was associated with a decrease in the prevalence of intestinal nematodes among newly arrived African and Southeast Asian refugees.


Paracrine mediation of calcium signaling in human SK-N-MCIXC neuroepithelioma cells.
Paracrine-mediated Ca2+ signaling in SK-N-MCIXC neuroepithelioma cells was evaluated by means of two experimental paradigms. In the first, single SK-N-MCIXC cells were microinjected with inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] and cytoplasmic Ca2+ was monitored by fura 2 digital-imaging microfluorometry. In response to Ins(1,4,5)P3 or CaCl2, but not inositol 1,3,4-trisphosphate, an increase in cytoplasmic Ca2+ concentration ([Ca2+]i) was observed in injected cells and also in neighboring cells. The direction of intercellular propagation of Ca2+ signals was influenced by the presence of a flow in the extracellular medium and occurred in the absence of any detectable gap-junctional communication. The P2 purinoceptor antagonist suramin, but not antagonists of other phosphoinositide-linked receptors, blocked cell-to-cell Ca2+ signaling initiated by microinjections of Ins(1,4,5)P3. In the second paradigm, conditioned medium (CM) obtained from monolayers of SK-N-MCIXC cells elicited increases in [Ca2+]i when reapplied to cells on coverslips. The Ca(2+)-mobilizing activity of CM was reversibly antagonized by suramin and abolished by pretreatment with apyrase. The presence of nucleotide di- and triphosphates in CM was confirmed by high-performance liquid chromatography. We conclude that SK-N-MCIXC cells release nucleotides that then activate specific receptors on neighboring cells. A rise in [Ca2+]i in these cells, and subsequent additional release of nucleotides, serves to further the propagation of intercellular Ca2+ signals.


Sea ice loss increases genetic isolation in a high Arctic ungulate metapopulation.
Sea ice loss may have dramatic consequences for population connectivity, extinction-colonization dynamics, and even the persistence of Arctic species subject to climate change. This is of particular concern in face of additional anthropogenic stressors, such as overexploitation. In this study, we assess the population-genetic implications of diminishing sea ice cover in the endemic, high Arctic Svalbard reindeer (Rangifer tarandus platyrhynchus) by analyzing the interactive effects of landscape barriers and reintroductions (following harvest-induced extirpations) on their metapopulation genetic structure. We genotyped 411 wild reindeer from 25 sampling sites throughout the entire subspecies' range at 19 microsatellite loci. Bayesian clustering analysis showed a genetic structure composed of eight populations, of which two were admixed. Overall population genetic differentiation was high (mean FST = 0.21). Genetic diversity was low (allelic richness [AR] = 2.07-2.58; observed heterozygosity = 0.23-0.43) and declined toward the outer distribution range, where populations showed significant levels of inbreeding. Coalescent estimates of effective population sizes and migration rates revealed strong evolutionary source-sink dynamics with the central population as the main source. The population genetic structure was best explained by a landscape genetics model combining strong isolation by glaciers and open water, and high connectivity by dispersal across winter sea ice. However, the observed patterns of natural isolation were strongly modified by the signature of past harvest-induced extirpations, subsequent reintroductions, and recent lack of sea ice. These results suggest that past and current anthropogenic drivers of metapopulation dynamics may have interactive effects on large-scale ecological and evolutionary processes. Continued loss of sea ice as a dispersal corridor within and between island systems is expected to increase the genetic isolation of populations, and thus threaten the evolutionary potential and persistence of Arctic wildlife.


Ethenoguanines undergo glycosylation by nucleoside 2'-deoxyribosyltransferases at non-natural sites.
Deoxyribosyl transferases and functionally related purine nucleoside phosphorylases are used extensively for synthesis of non-natural deoxynucleosides as pharmaceuticals or standards for characterizing and quantitating DNA adducts. Hence exploring the conformational tolerance of the active sites of these enzymes is of considerable practical interest. We have determined the crystal structure at 2.1 Å resolution of Lactobacillus helveticus purine deoxyribosyl transferase (PDT) with the tricyclic purine 8,9-dihydro-9-oxoimidazo[2,1-b]purine (N2,3-ethenoguanine) at the active site. The active site electron density map was compatible with four orientations, two consistent with sites for deoxyribosylation and two appearing to be unproductive. In accord with the crystal structure, Lactobacillus helveticus PDT glycosylates the 8,9-dihydro-9-oxoimidazo[2,1-b]purine at N7 and N1, with a marked preference for N7. The activity of Lactobacillus helveticus PDT was compared with that of the nucleoside 2'-deoxyribosyltransferase enzymes (DRT Type II) from Lactobacillus leichmannii and Lactobacillus fermentum, which were somewhat more effective in the deoxyribosylation than Lactobacillus helveticus PDT, glycosylating the substrate with product profiles dependent on the pH of the incubation. The purine nucleoside phosphorylase of Escherichia coli, also commonly used in ribosylation of non-natural bases, was an order of magnitude less efficient than the transferase enzymes. Modeling based on published active-site structures as templates suggests that in all cases, an active site Phe is critical in orienting the molecular plane of the purine derivative. Adventitious hydrogen bonding with additional active site residues appears to result in presentation of multiple nucleophilic sites on the periphery of the acceptor base for ribosylation to give a distribution of nucleosides. Chemical glycosylation of O9-benzylated 8,9-dihydro-9-oxoimidazo[2,1-b]purine also resulted in N7 and N1 ribosylation. Absent from the enzymatic and chemical glycosylations is the natural pattern of N3 ribosylation, verified by comparison of spectroscopic and chromatographic properties with an authentic standard synthesized by an unambiguous route.


Restricted replication of hepatitis A virus in cell culture: encapsidation of viral RNA depletes the pool of RNA available for replication.
The replication of hepatitis A virus (HAV) in BS-C-1 cells was examined under single-cycle growth conditions by using strand-specific probes for detection of viral RNA species. No measurable lag phase was demonstrated between accumulation of positive-strand HAV RNA and production of infectious virions, indicating that replication of virion RNA is rate limiting for the production of infectious virus. Intracellular viral RNA was further analyzed by using 2 M LiCl to fractionate the insoluble nonvirion 35S RNA and replicative intermediates (RI) from the soluble virions and double-stranded replicative forms, in conjunction with sucrose density gradient ultracentrifugation to separate the different forms of viral RNA. Throughout the productive phase of HAV infection, 95 to 97% of positive-strand HAV RNA was soluble in 2 M LiCl and was shown to be contained in mature virions. Of the LiCl-insoluble HAV RNA, more than 99% was positive-stranded 35S RNA, whereas 0.4% was negative stranded and had the sedimentation and partial RNase resistance characteristics of RI. The pattern of RNA accumulation in HAV-infected cells is thus very different from that seen in poliovirus-infected cells, where large pools of RI and mRNA are produced before RNA is sequestered into mature virions. The results of this study suggest that encapsidation of positive-strand HAV RNA inhibits transcription at all times during the growth cycle, thereby reducing the pool of replicating RNA and the final yield of infectious HAV.


En face imaging of single cell layers by differential phase-contrast optical coherence microscopy.
Optical coherence microscopy (OCM) is capable of imaging the backscattering potential of a sample with high transversal and axial resolution. We report on a combination of OCM with a differential phase-contrast technique that permits imaging of the subwavelength optical path differences that occur between a narrow beam probing a sample and its surrounding. This technique allows small transversal refractive-index variations close to a selected interface to be seen. We report on the method and present first images of a test sample and a single cell layer. The cells act as phase objects; imaging the phase properties improves the contrast compared with that of intensity images.


Distinguishing Loss of Structure from Subunit Dissociation for Protein Complexes with Variable Temperature Ion Mobility Mass Spectrometry.
The thermal stability and strength of interactions in proteins are commonly measured using isothermal calorimetry and differential scanning calorimetry providing a measurement that averages over structural transitions that occur as the proteins melt and dissociate. Here, we apply variable temperature ion mobility mass spectrometry (VT-IM-MS) to study the effect of temperature on the stability and structure of four multimeric protein complexes. VT-IM-MS is used here to investigate the change in the conformation of model proteins, namely, transthyretin (TTR), avidin, concanavalin A (conA), and human serum amyloid P component (SAP) at elevated temperatures prior, during, and after dissociation up to 550 K. As the temperature of the buffer gas is increased from 300 to 350 K, a small decrease in the collision cross sections ((DT)CCS(He)) of protein complexes from the values at room temperature is observed, and is associated with complex compaction occurring close to the reported solution T(m). At significantly higher temperatures, each protein complex undergoes an increase in (DT)CCS(He) and in the width of arrival time distributions (ATD), which is attributed to extensive protein unfolding, prior to ejection of a highly charged monomer species. This approach allows us to decouple the distinct gas phase melting temperature (T(m)) from the temperature at which we see subunit dissociation. The thermally induced dissociation (TID) mechanism is observed to initially proceed via the so-called "typical" (CID) dissociation route. Interestingly, data collected at higher analysis temperature suggests that the TID process might be adapting more "atypical" dissociation route.


Pharmacogenomics and pharmacoproteomics in the evaluation and management of short stature.
It has long been recognized that growth failure encompasses a diverse spectrum of underlying pathophysiological processes, a characteristic that has significantly impacted both the diagnosis and management of growth disorders. This problem is exacerbated by inherent difficulty in distinguishing the borders between the 'normal range' for stature and defined abnormal growth. Evaluation of GH secretion has proven problematic, both diagnostically and prognostically, except in cases of unequivocal GH deficiency. Measurement of serum concentrations of IGF-I, IGFBP-3, and ALS have proven useful in the assessment of GH responsiveness and have contributed to the concept of primary and secondary 'IGF deficiency'. Nevertheless, there is great need for biochemical and/or molecular biomarkers that could: i) predict short- and long-term responsiveness to various therapeutic modalities, such as GH and IGF-I, and ii) predict potential risk for adverse effects of therapy. Candidate proteins and genes identified to date, and worthy of further evaluation, include IGF-I, IGF-I receptor, GH receptor and its variants (such as exon 3-deleted GHR), STAT5b and short stature homeobox. Proteomic analysis of serum samples pre- and post-treatment and correlation with clinical responsiveness should provide additional candidate biomarkers. Molecular studies to consider include: i) sequencing and mutation analysis of known genetic components of the GH-IGF axis; ii) evaluation of single nucleotide polymorphisms of candidate genes; and iii) identification of new candidate genes. It is proposed that the major target population to study is that of children currently labeled as idiopathic short stature (ISS). These children can be divided into those with: i) primary IGFD, where the focus should be on genes related to GHR, GHR signaling, and IGF-I gene expression, or ii) no IGFD (i.e. 'true ISS'), where the focus should be on genes related to IGFR, IGF signaling and epiphyseal growth.


Neurophysiological, neuropsychological, and cognitive effects of 30 days of isolation.
The increasing demand of space flights requires a profound knowledge of the chronologic reactions of the human body to extreme conditions. Prior studies already have shown the adverse effects of long-term isolation on psycho-physiological well-being. The chronology of the effects and whether short-term isolation periods already lead to similar effects has not been investigated. Therefore, the aim of the current study was to investigate the effects of short-term isolation (30 days) on mood, cognition, cortisol, neurotrophic factors, and brain activity. 16 participants were isolated in the Human Exploration Research Analog at NASA for 30 days. 17 non-isolated control participants were tested simultaneously. On mission days - 5, 7, 14, 28, and + 5, multiple tests including the Positive and Negative Affect Schedule-X and cognitive tests were conducted, and a 5-min resting electroencephalography was recorded. A fasted morning blood drawing was also done. Increased stress was observed via augmented cortisol levels during the isolation period. Activity within the parietal cortex was reduced over time, probably representing a neural adaptation to less external stimuli. Cognitive performance was not affected, but rather enhanced in both groups. No further significant changes in neurotrophic factors BDNF/IGF-1 and mood could be detected. These results suggest that 30 days of isolation do not have a significant impact on brain activity, neurotrophic factors, cognition, or mood, even though stress levels were significantly increased during isolation. Further studies need to address the question as to what extent increased levels of stress do not affect mental functions during isolation periods.


The timing of anterior temporal lobe involvement in semantic processing.
Despite indications that regions within the anterior temporal lobe (ATL) might make a crucial contribution to pan-modal semantic representation, to date there have been no investigations of when during semantic processing the ATL plays a critical role. To test the timing of the ATL involvement in semantic processing, we studied the effect of double-pulse TMS on behavioral responses in semantic and difficulty-matched control tasks. Chronometric TMS was delivered over the left ATL (10 mm from the tip of the temporal pole along the middle temporal gyrus). During each trial, two pulses of TMS (40 msec apart) were delivered either at baseline (before stimulus presentation) or at one of the experimental time points 100, 250, 400, and 800 msec poststimulus onset. A significant disruption to performance was identified from 400 msec on the semantic task but not on the control assessment. Our results not only reinforce the key role of the left ATL in semantic representation but also indicate that its contribution is especially important around 400 msec poststimulus onset. Together, these facts suggest that the ATL may be one of the neural sources of the N400 ERP component.


Daytime pattern of post-exercise protein intake affects whole-body protein turnover in resistance-trained males.
The pattern of protein intake following exercise may impact whole-body protein turnover and net protein retention. We determined the effects of different protein feeding strategies on protein metabolism in resistance-trained young men. Participants were randomly assigned to ingest either 80g of whey protein as 8x10g every 1.5h (PULSE; n=8), 4x20g every 3h (intermediate, INT; n=7), or 2x40g every 6h (BOLUS; n=8) after an acute bout of bilateral knee extension exercise (4x10 repetitions at 80% maximal strength). Whole-body protein turnover (Q), synthesis (S), breakdown (B), and net balance (NB) were measured throughout 12h of recovery by a bolus ingestion of [15N]glycine with urinary [15N]ammonia enrichment as the collected end-product. PULSE Q rates were greater than BOLUS (~19%, P<0.05) with a trend towards being greater than INT (~9%, P=0.08). Rates of S were 32% and 19% greater and rates of B were 51% and 57% greater for PULSE as compared to INT and BOLUS, respectively (P<0.05), with no difference between INT and BOLUS. There were no statistical differences in NB between groups (P=0.23); however, magnitude-based inferential statistics revealed likely small (mean effect±90%CI; 0.59±0.87) and moderate (0.80±0.91) increases in NB for PULSE and INT compared to BOLUS and possible small increase (0.42±1.00) for INT vs. PULSE. We conclude that the pattern of ingested protein, and not only the total daily amount, can impact whole-body protein metabolism. Individuals aiming to maximize NB would likely benefit from repeated ingestion of moderate amounts of protein (~20g) at regular intervals (~3h) throughout the day.


An expanded model of replicating human immunodeficiency virus reverse transcriptase.
Replication complexes containing wild-type and RNase H-deficient p66/p51 human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT) were analyzed by DNase I and S1 footprinting. While crystallography and chemical footprinting data demonstrate that 15-18 bases of primer and template occupy the DNA polymerase and RNase H active centers, enzymatic footprinting suggests that a larger portion of substrate is encompassed by the replicating enzyme. Independent of the position of DNA synthesis arrest, template nucleotides +7 to -23 and primer nucleotides -1 to -25 are nuclease resistant. On both DNA strands, position -20 remains accessible to DNase I cleavage, suggestive of an alteration in nucleic acid structure between exiting the RNase H catalytic center and leaving the C-terminal p66 domain. A model of HIV-1 RT containing an extended single-stranded template and duplex region was constructed on the basis of the structure of an RT/DNA complex. Mapping of footprint data onto this model shows consistency between biochemical and structural data, implicating a contribution from domains proximal to the catalytic centers.


α-Synuclein in central nervous system and from erythrocytes, mammalian cells, and Escherichia coli exists predominantly as disordered monomer.
Since the discovery and isolation of α-synuclein (α-syn) from human brains, it has been widely accepted that it exists as an intrinsically disordered monomeric protein. Two recent studies suggested that α-syn produced in Escherichia coli or isolated from mammalian cells and red blood cells exists predominantly as a tetramer that is rich in α-helical structure (Bartels, T., Choi, J. G., and Selkoe, D. J. (2011) Nature 477, 107-110; Wang, W., Perovic, I., Chittuluru, J., Kaganovich, A., Nguyen, L. T. T., Liao, J., Auclair, J. R., Johnson, D., Landeru, A., Simorellis, A. K., Ju, S., Cookson, M. R., Asturias, F. J., Agar, J. N., Webb, B. N., Kang, C., Ringe, D., Petsko, G. A., Pochapsky, T. C., and Hoang, Q. Q. (2011) Proc. Natl. Acad. Sci. 108, 17797-17802). However, it remains unknown whether or not this putative tetramer is the main physiological form of α-syn in the brain. In this study, we investigated the oligomeric state of α-syn in mouse, rat, and human brains. To assess the conformational and oligomeric state of native α-syn in complex mixtures, we generated α-syn standards of known quaternary structure and conformational properties and compared the behavior of endogenously expressed α-syn to these standards using native and denaturing gel electrophoresis techniques, size-exclusion chromatography, and an oligomer-specific ELISA. Our findings demonstrate that both human and rodent α-syn expressed in the central nervous system exist predominantly as an unfolded monomer. Similar results were observed when human α-syn was expressed in mouse and rat brains as well as mammalian cell lines (HEK293, HeLa, and SH-SY5Y). Furthermore, we show that α-syn expressed in E. coli and purified under denaturing or nondenaturing conditions, whether as a free protein or as a fusion construct with GST, is monomeric and adopts a disordered conformation after GST removal. These results do not rule out the possibility that α-syn becomes structured upon interaction with other proteins and/or biological membranes.


Appropriate use of cancer comprehensive genome profiling assay using circulating tumor DNA.
Comprehensive genomic profiling (CGP) is being increasingly used for the routine clinical management of solid cancers. In July 2018, the use of tumor tissue-based CGP assays became available for all solid cancers under the universal health insurance system in Japan. Several restrictions presently exist, such as patient eligibility and limitations on the opportunities to perform such assays. The clinical implementation of CGP based on plasma circulating tumor DNA (ctDNA) is also expected to raise issues regarding the selection and use of tissue DNA and ctDNA CGP. A Joint Task Force for the Promotion of Cancer Genome Medicine comprised of three Japanese cancer-related societies has formulated a policy proposal for the appropriate use of plasma CGP (in Japanese), available at https://www.jca.gr.jp/researcher/topics/2021/files/20210120.pdf, http://www.jsco.or.jp/jpn/user_data/upload/File/20210120.pdf, and https://www.jsmo.or.jp/file/dl/newsj/2765.pdf. Based on these recommendations, the working group has summarized the respective advantages and cautions regarding the use of tissue DNA CGP and ctDNA CGP with reference to the advice of a multidisciplinary expert panel, the preferred use of plasma specimens over tissue, and multiple ctDNA testing. These recommendations have been prepared to maximize the benefits of performing CGP assays and might be applicable in other countries and regions.


The bidirectional association of 24-h activity rhythms and sleep with depressive symptoms in middle-aged and elderly persons.
In older populations disturbed 24-h activity rhythms, poor sleep, and depressive symptoms are often lingering and co-morbid, making treatment difficult. To improve insights into these commonly co-occurring problems, we assessed the bidirectional association of sleep and 24-h activity rhythms with depressive symptoms in middle-aged and elderly persons. In 1734 participants (mean age: 62.3 ± 9.3 years, 55% women) from the prospective Rotterdam Study, 24-h activity rhythms and sleep were estimated with actigraphy (mean duration: 146 ± 19.6 h), sleep quality with the Pittsburgh Sleep Quality Index, and depressive symptoms with the Center for Epidemiological Studies Depression scale. Repeated measures were available for 947 participants (54%) over a median follow-up of 6 years (interquartile range = 5.6-6.3). Linear-mixed models were used to assess temporal associations of 24-h activity rhythms and sleep with depressive symptoms in both directions. High 24-h activity rhythm fragmentation (IV) (B = 1.002, 95% confidence interval (CI) = 0.641-1.363), long time in bed (TIB) (B = 0.111, 95% CI = 0.053-0.169), low sleep efficiency (SE) (B = -0.015, 95% CI = -0.020 to -0.009), long sleep onset latency (SOL) (B = 0.009, 95% CI = 0.006-0.012), and low self-rated sleep quality (B = 0.112, 95% CI = 0.0992-0.124) at baseline were associated with increasing depressive symptoms over time. Conversely, more depressive symptoms at baseline were associated with an increasing 24-h activity rhythm fragmentation (B = 0.002, 95% CI = 0.001-0.003) and TIB (B = 0.009, 95% CI = 0.004-0.015), and a decreasing SE (B = -0.140, 95% CI = -0.196 to -0.084), SOL (B = 0.013, 95% CI = 0.008-0.018), and self-rated sleep quality (B = 0.193, 95% CI = 0.171-0.215) over time. This study demonstrates a bidirectional association of 24-h activity rhythms, actigraphy-estimated sleep, and self-rated sleep quality with depressive symptoms over a time frame of multiple years in middle-aged and elderly persons.


High-speed electrically actuated elastomers with strain greater than 100%
Electrical actuators were made from films of dielectric elastomers (such as silicones) coated on both sides with compliant electrode material. When voltage was applied, the resulting electrostatic forces compressed the film in thickness and expanded it in area, producing strains up to 30 to 40%. It is now shown that prestraining the film further improves the performance of these devices. Actuated strains up to 117% were demonstrated with silicone elastomers, and up to 215% with acrylic elastomers using biaxially and uniaxially prestrained films. The strain, pressure, and response time of silicone exceeded those of natural muscle; specific energy densities greatly exceeded those of other field-actuated materials. Because the actuation mechanism is faster than in other high-strain electroactive polymers, this technology may be suitable for diverse applications.


The frequent and underrecognised co-occurrence of acute exacerbated COPD and depression warrants screening: a systematic review.
Patients with acute exacerbated chronic obstructive pulmonary disease (AECOPD) and concurrent depression suffer significant psychological stress and decreased quality of life. The aim of this study was to collate data, guidelines and recommendations from publications on the screening and management of depressive mood disorders in patients hospitalised with AECOPD.We systematically searched four databases for publications reporting screening or management of depression in patients hospitalised for AECOPD. The identification of articles was based on the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement.Out of 1494 original articles screened, 35 met all inclusion criteria. These report a prevalence of depression in AECOPD ranging between 9.5% and 85.6%. Some studies report high postadmission mortality rates for depressive AECOPD patients, and higher readmission rates in depressive versus nondepressive AECOPD patients. Importantly, none of the 35 publications included suggestions on the screening and management of depression in AECOPD.Depression and AECOPD frequently co-occur, and this worsens outcomes. Yet we did not find recommendations on management, and few interventional studies. Patients hospitalised with AECOPD should be systematically screened for depression and treatment recommendations should be developed for these patients. Randomised studies on how to screen and treat depression in hospitalised AECOPD are necessary.


MYH7-related myopathies: clinical, histopathological and imaging findings in a cohort of Italian patients.
Myosin heavy chain 7 (MYH7)-related myopathies are emerging as an important group of muscle diseases of childhood and adulthood, with variable clinical and histopathological expression depending on the type and location of the mutation. Mutations in the head and neck domains are a well-established cause of hypertrophic cardiomyopathy whereas mutation in the distal regions have been associated with a range of skeletal myopathies with or without cardiac involvement, including Laing distal myopathy and Myosin storage myopathy. Recently the spectrum of clinical phenotypes associated with mutations in MYH7 has increased, blurring this scheme and adding further phenotypes to the list. A broader disease spectrum could lead to misdiagnosis of different congenital myopathies, neurogenic atrophy and other neuromuscular conditions. As a result of a multicenter Italian study we collected clinical, histopathological and imaging data from a population of 21 cases from 15 families, carrying reported or novel mutations in MYH7. Patients displayed a variable phenotype including atypical pictures, as dropped head and bent spine, which cannot be classified in previously described groups. Half of the patients showed congenital or early infantile weakness with predominant distal weakness. Conversely, patients with later onset present prevalent proximal weakness. Seven patients were also affected by cardiomyopathy mostly in the form of non-compacted left ventricle. Muscle biopsy was consistent with minicores myopathy in numerous cases. Muscle MRI was meaningful in delineating a shared pattern of selective involvement of tibialis anterior muscles, with relative sparing of quadriceps. This work adds to the genotype-phenotype correlation of MYH7-relatedmyopathies confirming the complexity of the disorder.


Perception of health risks among adolescents due to consumption of cigarettes, alcohol and psychoactive substances in the Federation of Bosnia and Herzegovina.
This article describes the perception of health risks in adolescents due to the consumption of cigarettes, alcohol and psychoactive substances in the Federation of Bosnia and Herzegovina (FBIH), as well as their observation of the behavior of their peers related to addictive behaviors. For the analysis was used a database from the European School Survey Project on Alcohol and Other Drugs (ESPAD) survey which was conducted in FBIH in 2011. The target population were students in the second grade of secondary schools in FBIH born in 1995 according to the ESPAD protocol. The total number of respondents from the cohort born in 1995 was 3813 students. The research results showed that the prevalence of risk perception due to the consumption of cigarettes, alcohol and psychoactive substances among adolescents in the FBIH is lower than the mean prevalence in countries which have implemented the ESPAD survey of 2011. PPreventive activities should be aimed at adolescent risk behaviours and empower them to make the right decisions that can have far reaching significance. Attention has to be paid to selective prevention that is directed towards individuals or subgroup of population where the risk of developing disorder is much higher than average.


Supporting tool suite for production proteomics.
The large amount of data produced by proteomics experiments requires effective bioinformatics tools for the integration of data management and data analysis. Here we introduce a suite of tools developed at Vanderbilt University to support production proteomics. We present the Backup Utility Service tool for automated instrument file backup and the ScanSifter tool for data conversion. We also describe a queuing system to coordinate identification pipelines and the File Collector tool for batch copying analytical results. These tools are individually useful but collectively reinforce each other. They are particularly valuable for proteomics core facilities or research institutions that need to manage multiple mass spectrometers. With minor changes, they could support other types of biomolecular resource facilities.


Resveratrol supplementation affects bone acquisition and osteoporosis: Pre-clinical evidence toward translational diet therapy.
Osteoporosis is a major public health issue that is expected to rise as the global population ages. Resveratrol (RES) is a plant polyphenol with various anti-aging properties. RES treatment of bone cells results in protective effects, but dose translation from in vitro studies to clinically relevant doses is limited since bioavailability is not taken into account. The aims of this review is to evaluate in vivo evidence for a role of RES supplementation in promoting bone health to reduced osteoporosis risk and potential mechanisms of action. Due to multiple actions on both osteoblasts and osteoclasts, RES has potential to attenuate bone loss resulting from different etiologies and pathologies. Several animal models have investigated the bone protective effects of RES supplementation. Ovariectomized rodent models of rapid bone loss due to estrogen-deficiency reported that RES supplementation improved bone mass and trabecular bone without stimulating other estrogen-sensitive tissues. RES supplementation prior to age-related bone loss was beneficial. The hindlimb unloaded rat model used to investigate bone loss due to mechanical unloading showed RES supplementation attenuated bone loss in old rats, but had inconsistent bone effects in mature rats. In growing rodents, RES increased longitudinal bone growth, but had no other effects on bone. In the absence of human clinical trials, evidence for a role of RES on bone heath relies on evidence generated by animal studies. A better understanding of efficacy, safety, and molecular mechanisms of RES on bone will contribute to the determination of dietary recommendations and therapies to reduce osteoporosis. This article is part of a Special Issue entitled: Resveratol: Challenges in translating pre-clinical findings to improved patient outcomes.


Transcriptome surveillance by selective termination of noncoding RNA synthesis.
Pervasive transcription of eukaryotic genomes stems to a large extent from bidirectional promoters that synthesize mRNA and divergent noncoding RNA (ncRNA). Here, we show that ncRNA transcription in the yeast S. cerevisiae is globally restricted by early termination that relies on the essential RNA-binding factor Nrd1. Depletion of Nrd1 from the nucleus results in 1,526 Nrd1-unterminated transcripts (NUTs) that originate from nucleosome-depleted regions (NDRs) and can deregulate mRNA synthesis by antisense repression and transcription interference. Transcriptome-wide Nrd1-binding maps reveal divergent NUTs at most promoters and antisense NUTs in most 3' regions of genes. Nrd1 and its partner Nab3 preferentially bind RNA motifs that are depleted in mRNAs and enriched in ncRNAs and some mRNAs whose synthesis is controlled by transcription attenuation. These results define a global mechanism for transcriptome surveillance that selectively terminates ncRNA synthesis to provide promoter directionality and to suppress antisense transcription.


Pretreatment platelet and hemoglobin levels are neither predictive nor prognostic variables for patients with locally advanced cervical cancer treated with neoadjuvant chemotherapy and radical hysterectomy: a retrospective Italian study.
To assess the predictive and prognostic values of pretreatment platelet and hemoglobin levels in patients with locally advanced cervical cancer who underwent neoadjuvant chemotherapy followed by radical hysterectomy. The authors retrospectively assessed 140 patients with FIGO (International Federation of Gynecology and Obstetrics) stage IB2-IIB cervical cancer who underwent chemosurgical treatment. Median pretreatment platelet and hemoglobin levels were 272,000/μL and 12.5 g/dL, respectively. Fourteen patients achieved a pathologically complete response, and 26 had an optimal partial response, with an optimal pathological response rate of 28.6%. By univariate analysis, optimal pathological response rate was associated with well/moderately differentiated grade (P = 0.02) and platinum-/paclitaxel-based chemotherapy regimen (P = 0.04), but not with platelet and hemoglobin levels. Multiple logistic regression confirmed that tumor grade (odds ratio, 2.827; 95% confidence interval [95% CI], 1.250-6.397; P = 0.01) and chemotherapy regimen (odds ratio, 5.416; 95% CI, 1.459-20.110; P = 0.01) were independent predictors of optimal pathological response. Five-year recurrence-free survival and 5-year overall survival were 66 % and 73%, respectively. By log-rank test, recurrence-free survival and overall survival were associated with pathological response (P = 0.0002 and P = 0.001), lymph node status, (P = 0.008 and P = 0.002), lymphovascular space status (P = 0.005 and P = 0.003), and parametrial and/or surgical margin status (P = 0.004 and P = 0.001), but not with platelet and hemoglobin levels. On multivariate analysis, pathological response (hazard ratio [HR], 7.999; 95% CI, 1.916-33.394; and HR, 6.007; 95% CI, 1.426-25.307) and parametrial and/or surgical margin status (HR, 2.061; 95%CI, 1.047-4.058; and HR, 2.561; 95% CI, 1.244-5.271) were independent prognostic variables of recurrence-free survival and overall survival. The achievement of an optimal pathological response is the strongest independent prognostic variable for patients with cervical cancer treated with neoadjuvant chemotherapy and radical hysterectomy, whereas pretreatment platelet and hemoglobin levels seem to be neither predictive of response to chemotherapy nor prognostic of long-term outcome.


Transflective display using a polymer-stabilized blue-phase liquid crystal.
A wide view, submillisecond response, and single-cell-gap transflective display using a blue-phase liquid crystal (BPLC) is proposed. To balance the optical phase retardation between transmissive (T) and reflective (R) regions, in-plane protrusion electrodes are formed with different gaps in the two regions. This display exhibits reasonably high optical efficiency and well matched voltage dependent transmittance and reflectance curves. Using biaxial films and broadband wide-view circular polarizers, the viewing angle with 100:1 contrast ratio is obtained over the entire viewing cone in the T region, and 10:1 over 50 degrees in the R region. The potential application is emphasized.


Radiostereometric analysis using clinical radiographic views: Validation measuring total hip replacement wear.
Radiostereometric analysis (RSA) is a stereo X-ray technique used in clinical research studies to evaluate micro-motion and wear of orthopaedic implants within bone. While highly accurate and precise, its adoption has been limited due to technical requirements such as the need for implanted marker beads and radiograph view angles determined by a calibration cage. We propose a new technique that separates the calibration procedure from the patient examination, enabling clinical radiograph views to be used for RSA measurements. The concept of a reference plate was adapted to establish the link between calibration procedure and patient examination procedure for cassette radiography, which may not be necessary for digital radiography. A hip wear phantom was used to validate this technique by comparing the error and repeatability of the novel procedure with that of conventional RSA. Femoral head penetration was measured versus the acetabular cup (head/cup) and marker beads in the acetabular liner (head/liner). Conventional RSA had lower inferior-superior average error (p = 0.03 for head/cup) while the modified RSA had lower anterior-posterior average error (p = 0.01). Average error was greater but not significantly so for the medial-lateral (p = 0.06) and 3D (p = 0.97) measurements. The head/liner method had lower average errors (p < 0.0001) for both procedures, but did not affect repeatability, which was similar between techniques. The novel procedure's average error and repeatability was therefore, similar to conventional RSA. This new technique could be applied to any joint with two clinical radiograph view angles pending further validation in subjects. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 34:1521-1528, 2016.


Transcranial magnetic stimulation of the posterior parietal cortex degrades accuracy of memory-guided saccades in humans.
Previous neurophysiological studies in the monkey have shown that the posterior parietal cortex (PPC) is involved in the control of memory-guided saccades. In this study, the authors investigated the effect of focal transcranial magnetic stimulation (TMS) over the posterior parietal cortex on accuracy of memory-guided saccades in humans to clarify the cortical region related to controlling memory-guided saccades. Single TMS pulses were applied systematically at various locations over the parietal cortex during memory-guided saccades in human subjects. Each subject was instructed to fixate on a central target until the target was extinguished. A lateral target was flashed for 100 msec with unpredictable direction and amplitude. The subject executed a saccade to the remembered position of the previously flashed target in total darkness when the central target was extinguished. Transcranial magnetic stimulation was applied after the extinction of the central target with various latencies. Stimulation sites were identified by three-dimensional magnetic resonance imaging (MRI). Transcranial magnetic stimulation of the right PPC degraded accuracy of rightward and leftward memory-guided saccades in all subjects, when TMS was applied 100 msec after the offset of the central target. On the other hand, TMS of the left PPC showed no effects on accuracy of memory-guided saccades. Systematic error of saccade amplitude appeared as a hypometria for rightward saccades. Three-dimensional MRI revealed that the effective stimulation site was located over the posterior portion of the intraparietal sulcus and the superior part of the angular gyrus of the right hemisphere. These findings strongly suggest that the right PPC is involved in maintaining spatial accuracy of remembered target locations of memory-guided saccades in humans.


C-reactive protein at discharge and 1-year mortality in hospitalised patients with acute decompensated heart failure: an observational study.
To examine the association of a high C-reactive protein (CRP) level at discharge from an acute decompensated heart failure (ADHF) hospitalisation with the 1-year clinical outcomes. A post-hoc subanalysis of a prospective cohort study of patients hospitalised for ADHF (using the Kyoto Congestive Heart Failure (KCHF) registry) between October 2014 and March 2016 with a 1-year follow-up. A physician-initiated multicentre registry enrolled consecutive hospitalised patients with ADHF for the first time at 19 secondary and tertiary hospitals in Japan. Among the 4056 patients enrolled in the KCHF registry, the present study population consisted of 2618 patients with an available CRP value both on admission and at discharge and post-discharge clinical follow-up data. We divided the patients into two groups, those with a high CRP level (>10 mg/L) and those with a low CRP level (≤10 mg/L) at discharge from the index hospitalisation. The primary outcome measure was all-cause death after discharge from the index hospitalisation. The secondary outcome measures were heart failure hospitalisations, cardiovascular death and non-cardiovascular death. The high CRP group and low CRP group included 622 patients (24%) and 1996 patients (76%), respectively. During a median follow-up period of 468 days, the cumulative 1-year incidence of the primary outcome was significantly higher in the high CRP group than low CRP group (24.1% vs 13.9%, log-rank p<0.001). Even after a multivariable analysis, the excess mortality risk in the high CRP group relative to the low CRP group remained significant (HR, 1.43; 95% CI, 1.19 to 1.71; p<0.001). The excess mortality risk was consistent regardless of the clinically relevant subgroup factors. A high CRP level (>10 mg/L) at discharge from an ADHF hospitalisation was associated with an excess mortality risk at 1 year. TRIAL REGISTRATION DETAILS: https://clinicaltrials.gov/ct2/show/NCT02334891 (NCT02334891) https://upload.umin.ac.jp/cgi-open-bin/ctr_e/ctr_view.cgi?recptno=R000017241 (UMIN000015238).


Seed size influence on germination responses to light and temperature of seven pioneer tree species from the Central Amazon.
In Amazon secondary forests are dominated by pioneer species that typically produce large amounts of small and dormant seeds that are able to form a persistent soil seed bank. Seed dormancy in this group of species is overcome by environmental conditions found in open areas, such as high irradiation or alternating temperatures. Nevertheless, a variety of germination responses to environmental factors is known among pioneers; some of them may germinate in diffuse light or in darkness condition at constant temperature. Seed mass can be considered as one of the factors that promotes this variety. Regarding species with very small seeds, it seems that the trigger for germination is light and for larger seeds temperature alternation may be a more important stimulus. In this study we established a relationship between seed mass and germination response to light and alternating temperature for a group of seven woody pioneer species from the Amazon forest. We found that an increase in seed mass was followed by a decrease in the need for light and an increase in the tolerance to alternating temperatures. Understanding germination strategies may contribute with the knowledge of species coexistence in high diverse environments and also may assist those involved in forest management and restoration.


A stable niche assumption-free test of ecological divergence.
Understanding the impact of geological events on diversification processes is central to evolutionary ecology. The recent amalgamation between ecological niche models (ENMs) and phylogenetic analyses has been used to estimate historical ranges of modern lineages by projecting current ecological niches of organisms onto paleoclimatic reconstructions. A critical assumption underlying this approach is that niches are stable over time. Using Notophthalmus viridescens (eastern newt), in which four ecologically diverged subspecies are recognized, we introduce an analytical framework free from the niche stability assumption to examine how refugial retreat and subsequent postglacial expansion have affected intraspecific ecological divergence. We found that the current subspecies designation was not congruent with the phylogenetic lineages. Thus, we examined ecological niche overlap between the refugial and modern populations, in both subspecies and lineage, by creating ENMs independently for modern and estimated last glacial maximum (LGM) newt populations, extracting bioclimate variables by randomly generated points, and conducting principal component analyses. Our analyses consistently showed that when tested as a hypothesis, rather than used as an assumption, the niches of N. viridescens lineages have been unstable since the LGM (both subspecies and lineages). There was greater ecological niche differentiation among the subspecies than the modern phylogenetic lineages, suggesting that the subspecies, rather than the phylogenetic lineages, is the unit of the current ecological divergence. The present study found little evidence that the LGM refugial retreat caused the currently observed ecological divergence and suggests that ecological divergence has occurred during postglacial expansion to the current distribution ranges.


PEGylated Liposomes as Carriers of Hydrophobic Porphyrins.
Sterically stabilized liposomes (SSLs) (PEGylated liposomes) are applied as effective drug delivery vehicles. Understanding the interactions between hydrophobic compounds and PEGylated membranes is therefore important to determine the effectiveness of PEGylated liposomes for delivery of drugs or other bioactive substances. In this study, we have combined fluorescence quenching analysis (FQA) experiments and all-atom molecular dynamics (MD) simulations to study the effect of membrane PEGylation on the location and orientation of 5,10,15,20-tetrakis(4-hydroxyphenyl)porphyrin (p-THPP) that has been used in our study as a model hydrophobic compound. First, we consider the properties of p-THPP in the presence of different fluid phosphatidylcholine bilayers that we use as model systems for protein-free cell membranes. Next, we studied the interaction between PEGylated membranes and p-THPP. Our MD simulation results indicated that the arrangement of p-THPP within zwitterionic membranes is dependent on their free volume, and p-THPP solubilized in PEGylated liposomes is localized in two preferred positions: deep within the membrane (close to the center of the bilayer) and in the outer PEG corona (p-THPP molecules being wrapped with the polymer chains). Fluorescence quenching methods confirmed the results of atomistic MD simulations and showed two populations of p-THPP molecules as in MD simulations. Our results provide both an explanation for the experimental observation that PEGylation improves the drug-loading efficiency of membranes and also a more detailed molecular-level description of the interactions between porphyrins and lipid membranes.


Possible false-negative results on therapeutic drug monitoring of phenytoin using a particle enhanced turbidimetric inhibition immunoassay in a patient with a high level of IgM.
: In this report, the authors described the unusual case of a patient in whom the plasma phenytoin concentration was unexpectedly not detected on a particle-enhanced turbidimetric inhibition immunoassay (PETINIA) technique, a typical immunoassay for phenytoin. The plasma concentration was measured using PETINIA and high-performance liquid chromatography in a 69-year-old male patient treated with fosphenytoin intravenously at the standard dose for 7 days. Although the plasma concentration of phenytoin was below the limit of detection (<0.5 mcg/mL) on PETINIA after the administration of fosphenytoin, the trough plasma concentration was estimated to be between 5 and 10 mg/L on high-performance liquid chromatography. When the plasma concentrations of IgM and IgG were measured using an enzyme-linked immunosorbent assay, the plasma IgG level was within the reference range, whereas the plasma IgM level was 2-3 times higher than the upper limit of the reference range. We concluded that the PETINIA method yielded a possible false-negative result regarding the phenytoin level in this patient, perhaps because of some hindrance to the measurement process by IgM. This case suggests that false-negative results should be considered when therapeutic drug monitoring reveals abnormally low values using PETINIA and that it is necessary to evaluate the plasma IgM level.


Focus on an unusual rise in pancreatic cancer incidence in France.
Pancreatic cancer is one of the most lethal. Most countries have exhibited a stable or decreasing incidence over time. The aim of this study was to provide updated French temporal trends in pancreatic cancer incidence and mortality over the past three decades. Incidence was estimated using the French National Network of Cancer Registries (FRANCIM) and mortality using the French Mortality Statistics Office. World age-standardized incidence and mortality were modelled by age-period-cohort models. The net cumulative risk of developing pancreatic cancer by birth cohort was calculated, as were annual percentage changes (APCs) in incidence and mortality. Between 1982 and 2012, age-standardized incidence increased from 4.8 in 1980 to 9.6 per 100 000 in men and from 2.3 to 6.8 in women. The mean APC was 2.3% (2.1-2.6) and 3.6% (3.3-3.9), respectively. The cumulative risk of developing pancreatic cancer before age 75 rose from 0.62% for males born around 1920 to 1.17% for those born around 1950. It was respectively 0.31% and 0.86% for women. Mortality did not vary in men (8.1 per 100 000). It slightly increased in women from 4.0 in 1982 to 5.4 in 2012. Pancreatic cancer incidence and mortality exhibited diverging trends. Incidence increased over the last 30 years in France whereas mortality did not vary in men and moderately increased in women. Incidence remained lower than mortality up to 2002. One cannot exclude the possibility that a similar trend may appear in other countries. Etiological studies are required to further explain this increase.


Surgical Management of Hidradenitis Suppurativa: Outcomes of 590 Consecutive Patients.
Hidradenitis suppurativa is a progressive, recurrent inflammatory disease. Surgical management is potentially curative with limited efficacy data. To evaluate hidradenitis surgical patients. Retrospective review of outcomes of 590 consecutive surgically treated patients. Most patients were white (91.0% [435/478]), men (337 [57.1%]), smokers (57.7% [297/515]) with Hurley Stage III disease (476 [80.7%]). Procedure types were excision (405 [68.6%]), unroofing (168 [28.5%]), and drainage (17 [2.9%]) treating disease of perianal/perineum (294 [49.8%]), axilla (124 [21.0%]), gluteal cleft (76 [12.9%]), inframammary (12 [2.0%]), and multiple surgical sites (84 [14.2%]). Postoperative complications occurred in 15 patients (2.5%) and one-fourth (144 [24.4%]) suffered postoperative recurrence, which necessitated reoperation in one-tenth (69 [11.7%]) of patients. Recurrence risk was increased by younger age (hazard ratio [HR], 0.8; 95% confidence interval [CI], 0.7-0.9), multiple surgical sites (HR, 1.6; 95% CI, 1.1-2.5), and drainage-type procedures (HR, 3.5; 95% CI, 1.2-10.7). Operative location, disease severity, gender, and operative extent did not influence recurrence rate. Excision and unroofing procedures were effective treatments with infrequent complications and low recurrence rates. Well-planned surgical treatment aiming to remove or unroof the area of intractable hidradenitis suppurativa was highly effective in the management of this challenging disease.


Mammalian target of rapamycin inhibitors permit regulatory T cell reconstitution and inhibit experimental chronic graft-versus-host disease.
Chronic graft-versus-host disease (GVHD) remains a major late complication of allogeneic bone marrow transplantation (BMT). In a previous study, impaired thymic negative selection of the recipients permitted the emergence of pathogenic T cells that cause chronic GVHD using MHC class II-deficient (H2-Ab1 KO) B6 into C3H model and CD4(+) T cells isolated from chronic GVHD mice caused chronic GVHD when administered into the secondary recipients. In this study, we evaluated the kinetics of regulatory T cell (Treg) reconstitution in wild type B6 into C3H model. After myeloablative conditioning, host Tregs disappeared rapidly, followed by expansion of Tregs derived from the donor splenic T cell inoculum. However, the donor splenic T cell-derived Treg pool contracted gradually and was almost completely replaced by newly generated donor bone marrow (BM)-derived Tregs in the late post-transplantation period. Next, we compared the effects of cyclosporine (CSA) and mammalian target of rapamycin (mTOR) inhibitors on Treg reconstitution. Administration of CSA significantly impaired Treg reconstitution in the spleen and thymus. In contrast, BM-derived Treg reconstitution was not impaired in mTOR inhibitor-treated mice. Histopathological examination indicated that mice treated with CSA, but not mTOR inhibitors, showed pathogenic features of chronic GVHD on day 120. Mice treated with CSA until day 60, but not mTOR inhibitors, developed severe chronic GVHD followed by adoptive transfer of the pathogenic CD4(+) T cells isolated from H2-Ab1 KO into C3H model. These findings indicated that long-term use of CSA impairs reconstitution of BM-derived Tregs and increases the liability to chronic GVHD. The choice of immunosuppression, such as calcineurin inhibitor-free GVHD prophylaxis with mTOR inhibitor, may have important implications for the control of chronic GVHD after BMT.


Dark age of sourcing cadavers in developing countries: a Nigerian survey.
Throughout the history of medical science, the sourcing, storage, and management of cadavers for the study of gross anatomy have been problematic issues. As presented in this report, a study of these issues was conducted in the departments of anatomy of medical schools in Nigeria. To establish the extent and depth of the prevailing difficulties in the sourcing of cadavers, 18 (62.1%) of the Departments of Anatomy that qualified for this study were used. In sum, 94.4% of these schools reported an inadequate number of cadavers for study. The estimated ages of the cadavers were between 20 and 40 years and 85% of the schools reported more than 75% of the cadavers to be from the lower socioeconomic class. Altogether, 94.4% of the schools reported a male dominance of more than 95%. More than 72% of the schools reported that more than 90% of the cadavers were criminals that had been killed by shooting, less than 10% were unclaimed and unidentified corpses, and 0% originated from body bequest. No form of screening exercises for diseases and infective microorganisms was noted for all the schools, and there were no set standards for the acceptance or rejection of cadavers. Causative factors for the profile of available cadavers such as culture and religion were noted. This study was designed to look at various issues associated with the sourcing of cadavers for Anatomy education in typical developing African countries like Nigeria. As outlined in this report, the creation of legislation and the promotion and funding of programs highlighting the importance of body donation are crucial for improving the field of medical education.


Metabolic and hypoxic adaptation to anti-angiogenic therapy: a target for induced essentiality.
Anti-angiogenic therapy has increased the progression-free survival of many cancer patients but has had little effect on overall survival, even in colon cancer (average 6-8 weeks) due to resistance. The current licensed targeted therapies all inhibit VEGF signalling (Table 1). Many mechanisms of resistance to anti-VEGF therapy have been identified that enable cancers to bypass the angiogenic blockade. In addition, over the last decade, there has been increasing evidence for the role that the hypoxic and metabolic responses play in tumour adaptation to anti-angiogenic therapy. The hypoxic tumour response, through the transcription factor hypoxia-inducible factors (HIFs), induces major gene expression, metabolic and phenotypic changes, including increased invasion and metastasis. Pre-clinical studies combining anti-angiogenics with inhibitors of tumour hypoxic and metabolic adaptation have shown great promise, and combination clinical trials have been instigated. Understanding individual patient response and the response timing, given the opposing effects of vascular normalisation versus reduced perfusion seen with anti-angiogenics, provides a further hurdle in the paradigm of personalised therapeutic intervention. Additional approaches for targeting the hypoxic tumour microenvironment are being investigated in pre-clinical and clinical studies that have potential for producing synthetic lethality in combination with anti-angiogenic therapy as a future therapeutic strategy.


Resistance to TRAIL in non-transformed cells is due to multiple redundant pathways.
Tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) is a cytokine and a selective inducer of apoptosis in a range of tumour cells, but not in normal, untransformed cells. A large number of chemotherapeutics as well as biological agents are being tested for their potential to sensitise resistant tumour cells to TRAIL as a means to broaden the range of tumours treatable with TRAIL. However, because of the incomplete understanding of the mechanism(s) underlying TRAIL resistance in non-malignant cells, it is unpredictable whether the effect of these sensitisers will be restricted to tumour cells or they would also sensitise non-transformed cells causing unwanted toxicity. In this study, we carried out a systematic analysis of the mechanisms driving TRAIL resistance in non-transformed cells. We found that cellular FLICE-like inhibitory protein, anti-apoptotic B-cell lymphoma 2 proteins, and X-linked inhibitor of apoptosis protein were independently able to provide resistance to TRAIL. Deficiency of only one of these proteins was not sufficient to elicit TRAIL sensitivity, demonstrating that in non-transformed cells multiple pathways control TRAIL resistance and they act in a redundant manner. This is contrary to the resistance mechanisms found in tumour cell types, many of them tend to rely on a single mechanism of resistance. Supporting this notion we found that 76% of TRAIL-resistant cell lines (13 out of 17) expressed only one of the above-identified anti-apoptotic proteins at a high level (≥1.2-fold higher than the mean expression across all cell lines). Furthermore, inhibition or knockdown of the single overexpressed protein in these tumour cells was sufficient to trigger TRAIL sensitivity. Therefore, the redundancy in resistance pathways in non-transformed cells may offer a safe therapeutic window for TRAIL-based combination therapies where selective sensitisation of the tumour to TRAIL can be achieved by targeting the single non-redundant resistance pathway.


Incidental and clinically actionable genetic variants in 1005 whole exomes and genomes from Qatar.
Incidental findings in genomic data have been studied in great detail in the recent years, especially from population-scale data sets. However, little is known about the frequency of such findings in ethnic groups, specifically the Middle East, which were not previously covered in global sequencing studies. The availability of whole exome and genome data sets for a highly consanguineous Arab population from Qatar motivated us to explore the incidental findings in this population-scale data. The sequence data of 1005 Qatari individuals were systematically analyzed for incidental genetic variants in the 59 genes suggested by the American College of Medical Genetics and Genomics. We identified four genetic variants which were pathogenic or likely pathogenic. These variants occurred in six individuals, suggesting a frequency of 0.59% in the population, much lesser than that previously reported from European and African populations. Our analysis identified a variant in RYR1 gene associated with Malignant Hyperthermia that has significantly higher frequency in the population compared to global frequencies. Evaluation of the allele frequencies of these variants suggested enrichment in sub-populations, especially in individuals of Sub-Saharan African ancestry. The present study thereby provides the information on pathogenicity and frequency, which could aid in genomic medicine. To the best of our knowledge, this is the first comprehensive analysis of incidental genetic findings in any Arab population and suggests ethnic differences in incidental findings.


Signal-to-noise-ratio analysis for nonlinear N-ary phase filters.
The problem of recognizing targets in nonoverlapping clutter using nonlinear N-ary phase filters is addressed. Using mathematical analysis, expressions were derived for an N-ary phase filter and the intensity variance of an optical correlator output. The N-ary phase filter was shown to consist of an infinite sum of harmonic terms whose periodicity was determined by N. For the intensity variance, it was found that under certain conditions the variance was minimized due to a previously undiscovered phase quadrature effect. Comparison showed that optimal real filters produced greater signal-to-noise-ratio values than the continuous phase versions as a consequence of this effect.


Reasons for choosing occupational therapy as a profession: implications for recruitment.
This study examines the ways in which 272 recently certified occupational therapists learned of the profession of occupational therapy and the factors that influenced their decisions to choose occupational therapy as a career. The data were analyzed according to age, sex, and geographic region to identify potential ways to improve recruitment efforts. The results indicated that therapists most often learned of the profession while working in a health care setting. First exposure to occupational therapy was usually through an acquaintance in the profession, a family member, or a friend. The primary reasons for choosing occupational therapy included a desire to help people with disabilities, the promise of challenge and variety, the opportunity to work in a health care setting, and the belief that jobs are plentiful. The factors most likely to dissuade people from the profession were the expense of an education and lack of a clear understanding of occupational therapy.


Dysregulation of mitochondrial bioenergetics and quality control by HIV-1 Tat in cardiomyocytes.
Cardiovascular disease remains a leading cause of morbidity and mortality in HIV-positive patients, even in those whose viral loads are well controlled with antiretroviral therapy. However, the underlying molecular events responsible for the development of cardiac disease in the setting of HIV remain unknown. The HIV-encoded Tat protein plays a critical role in the activation of HIV gene expression and profoundly impacts homeostasis in both HIV-infected cells and uninfected cells that have taken up released Tat via a bystander effect. Since cardiomyocyte function, including excitation-contraction coupling, greatly depends on energy provided by the mitochondria, in this study, we performed a series of experiments to assess the impact of Tat on mitochondrial function and bioenergetics pathways in a primary cell culture model derived from neonatal rat ventricular cardiomyocytes (NRVCs). Our results show that the presence of Tat in cardiomyocytes is accompanied by a decrease in oxidative phosphorylation, a decline in the levels of ATP, and an accumulation of reactive oxygen species (ROS). Tat impairs the uptake of mitochondrial Ca2+ ([Ca2+ ]m ) and the electrophysiological activity of cardiomyocytes. Tat also affects the protein clearance pathway and autophagy in cardiomyocytes under stress due to hypoxia-reoxygenation conditions. A reduction in the level of ubiquitin along with dysregulated degradation of autophagy proteins including SQSTM1/p62 and a reduction of LC3 II were detected in cardiomyocytes harboring Tat. These results suggest that, by targeting mitochondria and protein quality control, Tat significantly impacts bioenergetics and autophagy resulting in dysregulation of cardiomyocyte health and homeostasis.


Comparing 16S rDNA amplicon sequencing and hybridization capture for pea aphid microbiota diversity analysis.
Targeted sequencing of 16S rDNA amplicons is routinely used for microbial community profiling but this method suffers several limitations such as bias affinity of universal primers and short read size. Gene capture by hybridization represents a promising alternative. Here we used a metagenomic extract from the pea aphid Acyrthosiphon pisum to compare the performances of two widely used PCR primer pairs with DNA capture, based on solution hybrid selection. All methods produced an exhaustive description of the 8 bacterial taxa known to be present in this sample. In addition, the methods yielded similar quantitative results, with the number of reads strongly correlating with quantitative PCR controls. Both methods can thus be considered as qualitatively and quantitatively robust on such a sample with low microbial complexity.


First Report of Pythium recalcitrans Causing Cavity Spot of Carrot in Michigan.
During a survey of carrot (Daucus carota L.) cavity spot in Michigan in September 2010, carrot roots with typical cavity spot symptoms were collected from production fields in Fremont Co. The lesions were excised from infected roots, surface-disinfested with 0.62% NaClO for 3 min, rinsed in sterilized, distilled water three times, cut into 0.5 cm long pieces, and then plated on water agar (WA) amended with carbendazim (10 μg/ml), ampicillin (50 μg/ml), rifampicin (50 μg/ml), and pentachloronitrobenzene (10 μg/ml) (cumulatively referred to as CARP). Plates were incubated at 22 ± 1°C in the dark for 3 days. Pure cultures of the isolates were obtained by transferring a single hyphal tip of each colony to potato dextrose agar (PDA) amended with CARP. Among the 33 isolates obtained, M2-05 was identified as a Pythium sp. that differed from the known cavity spot pathogens of carrot. The isolate has spherical hyphal swellings but no other distinguishing morphological characteristics. M2-05 was further classified by analyzing the partial sequences of four genes: the internal transcribed spacer (ITS) region of ribosomal DNA, beta-tubulin (β-tub), cytochrome c oxidase subunit 2 (cox 2), and NADH dehydrogenase subunit 1 (nadh 1) (1,3). A BLAST search of these sequences for M2-05 was conducted using the nucleotide database of GenBank, resulting in 100% similarity to all four sequenced genes of P. recalcitrans (2). The DNA sequences of M2-05 were deposited in GenBank (JQ734349, JQ734229, JQ734289, and JQ734409 for ITS, β-tub, cox 2, and nadh 1, respectively). Koch's postulates were conducted by inoculating mature carrot roots (cv. Nantindo) with mycelial plugs (4 mm in diameter) cut from the margin of actively growing colonies of M2-05 on PDA plates. Two mycelial plugs were placed on each carrot root at 3-cm intervals, with the mycelial side facing the root; and two non-colonized agar plugs were placed similarly for the non-inoculated control treatment. In comparison, carrot roots also were inoculated with an isolate of each of P. sulcatum and P. violae using the same method. There were four replicate carrot roots inoculated for each isolate and each of the control treatments. The inoculated roots were placed on a plastic grid (7 mm in height) in a closed plastic container, with moist paper towels underneath the grids. The container was incubated in the dark at 22 ± 1°C, and the roots were sprayed gently daily with sterilized, distilled water to maintain high humidity. Brown lesions were observed on all inoculated carrot roots 5 days after inoculation. The lesions measured 0.68 ± 0.48, 1.20 ± 0.71, and 0.56 ± 0.31 mm2 averaged over all eight lesions for the isolates of P. recalcitrans, P. sulcatum, and P. violae, respectively. Symptomatic tissues from the inoculated roots were excised and incubated on WA-CARP plates, and the culture from each lesion confirmed as the isolates inoculated using the same molecular methods described above. The carrot tissue under the control agar plugs remained symptomless, and no Pythium was recovered from the control roots. P. recalcitrans was described in 2008 as infecting roots of Beta vulgaris and Vitis vinifera (2). To our knowledge, this is the first published report of P. recalcitrans naturally infecting carrot roots, not only in Michigan, but anywhere in the world. References: (1) L. Kroon et al. Fungal. Genet. Biol. 41:766, 2004. (2) E. Moralejo et al. Mycologia 100:310, 2008. (3) N. O. Villa et al. Mycologia 98:410, 2006.


Relationship between tobacco control policies and the delivery of smoking cessation services in nonprofit HMOs.
This project examined tobacco policies and delivery of cessation services in nonprofit HMOs that collectively provide comprehensive medical care to more than 8 million members. Three annual surveys with health plan managers showed that all of these health plans had written tobacco control guidelines that became more comprehensive over the span of this study. We also surveyed a random sample of 4207 current smokers who had attended a primary care visit in the past year (399-528 at each of nine health plans). Of these smokers, 71% reported advice to quit, 56% were asked about their willingness to quit, 49% were provided some assistance in quitting (mostly self-help material or information about classes or counseling), and 9% were offered some kind of follow-up. Smokers receiving assistance in quitting reported higher satisfaction with their care. In general, health plans with the most comprehensive policies also showed higher rates of implementing tobacco treatment programs in primary care. Compared with tobacco control efforts of a decade or more ago, considerable progress has been made. However, there is still room for improvement in the proportion of smokers who receive the most effective forms of assistance in quitting.


Direct observation approach for detecting medication errors and adverse drug events in a pediatric intensive care unit.
To determine the incidence, type, and stage of occurrence of medication errors and potential and actual adverse drug events (ADEs) in a pediatric intensive care unit (ICU) using trained observers. The preventability and severity of ADEs and the system failures leading to medication error occurrence were also investigated. Prospective, direct observation study. A 16-bed pediatric medical/surgical ICU at a tertiary care academic medical center. One enrolled nurse caring for at least one pediatric ICU patient age <18 yrs was randomly chosen during each observation period. Observers would intervene only in the event that the medication error would cause substantial patient harm or discomfort. Medication errors and potential and actual ADEs were identified throughout the entire medication use process. The 26 12-hr observation periods included 357 reviewed written orders and 263 observed doses. The study observers identified 58 incidents, which were subsequently classified by the evaluators according to clinical importance, severity, and preventability. Fifty-two of these incidents were considered medication errors; six incidents were determined to be nonpreventable ADEs. Of the 52 medication errors, 42 (81%) were considered clinically important. Potential ADEs comprised 35 (83%) of these important medication errors; the other seven (17%) were classified as actual, preventable ADEs. Overall, the actual and potential ADE rate occurred at 3.6 events and 9.8 events per 100 orders, respectively. Our medication error rate was similar to that of previous pediatric ICU studies that used the direct observation method for reporting but higher than the rates in previous studies using other detection techniques such as voluntary incident reporting. Periodic direct observation and other ongoing data collection methods such as voluntary incident reporting have the potential to be complementary approaches to medication error and ADE detection.


Strategies for monitoring and updating clinical practice guidelines: a systematic review.
Scientific knowledge is in constant change. The flow of new information requires a frequent re-evaluation of the available research results. Clinical practice guidelines (CPGs) are not exempted from this phenomenon and need to be kept updated to maintain the validity of their recommendations. The objective of our review is to systematically identify, describe and assess strategies for monitoring and updating CPGs. We conducted a systematic review of studies evaluating one or more methods of updating (with or without monitoring) CPGs or recommendations. We searched MEDLINE (PubMed) and The Cochrane Methodology Register (The Cochrane Library) from 1966 to June 2012. Additionally, we hand-searched reference lists of the included studies and the Guidelines International Network book of abstracts. If necessary, we contacted study authors to obtain additional information. We included a total of eight studies. Four evaluated if CPGs were out of date, three updated CPGs, and one continuously monitored and updated CPGs. The most detailed reported phase of the process was the identification of new evidence. As opposed to studies updating guidelines, studies evaluating if CPGs were out of date applied restricted searches. Only one study compared a restricted versus an exhaustive search suggesting that a restricted search is sufficient to assess recommendations' Validity. One study analyzed the survival time of CPGs and suggested that these should be reassessed every three years. There is limited evidence about the optimal strategies for monitoring and updating clinical practice guidelines. A restricted search is likely to be sufficient to monitor new evidence and assess the need to update, however, more information is needed about the timing and type of search. Only the exhaustive search strategy has been assessed for the update of CPGs. The development and evaluation of more efficient strategies is needed to improve the timeliness and reduce the burden of maintaining the validity of CPGs.


Authoritative subspecies diagnosis tool for European honey bees based on ancestry informative SNPs.
With numerous endemic subspecies representing four of its five evolutionary lineages, Europe holds a large fraction of Apis mellifera genetic diversity. This diversity and the natural distribution range have been altered by anthropogenic factors. The conservation of this natural heritage relies on the availability of accurate tools for subspecies diagnosis. Based on pool-sequence data from 2145 worker bees representing 22 populations sampled across Europe, we employed two highly discriminative approaches (PCA and FST) to select the most informative SNPs for ancestry inference. Using a supervised machine learning (ML) approach and a set of 3896 genotyped individuals, we could show that the 4094 selected single nucleotide polymorphisms (SNPs) provide an accurate prediction of ancestry inference in European honey bees. The best ML model was Linear Support Vector Classifier (Linear SVC) which correctly assigned most individuals to one of the 14 subspecies or different genetic origins with a mean accuracy of 96.2% ± 0.8 SD. A total of 3.8% of test individuals were misclassified, most probably due to limited differentiation between the subspecies caused by close geographical proximity, or human interference of genetic integrity of reference subspecies, or a combination thereof. The diagnostic tool presented here will contribute to a sustainable conservation and support breeding activities in order to preserve the genetic heritage of European honey bees.


The Effects of 8-Week Plyometric Training on Physical Performance in Young Tennis Players.
The aim of this study was to analyze the effects of an 8-week (conducted biweekly for a total of 16 sessions) plyometric training program (PT) (e.g., upper- and lower-body exercises) combined with regular tennis training on physical qualities in young tennis players. Sixty tennis players between the ages of 12 and 13 years (age 12.5 ± 0.3 years, weight 44.2 ± 7.0 kg, height 156.6 ± 7.1 cm) were allocated to either the control group (standard in-season regimen) (CG; n = 30) or the experimental group, which received an additional PT (TG; n = 30) for 30-60 min as a substitute for some tennis training within the usual 90-min practice. Pre- and posttests included: anthropometric measures; vertical countermovement jump (CMJ); standing long jump (SLJ); 20 m sprint time (with 5 and 10 m splits); a modified 505 agility test; overhead medicine ball throw; and serve velocity test. After the training intervention, the TG showed significant (p < .01) improvements in all the parameters analyzed, with percentages of change and effect sizes ranging from 3.1% to 10.1% and 0.4 (small) to 1.3 (moderate), respectively. No significant changes were observed in the CG after the training intervention. PT was shown as an important stimulus for enhancing explosive actions in young tennis players.


Evaluation of late outgrowth endothelial progenitor cell and umbilical vein endothelial cell responses to thromboresistant collagen-mimetic hydrogels.
Bioactive coatings which support the adhesion of late-outgrowth peripheral blood endothelial progenitor cells (EOCs) are actively being investigated as a means to promote rapid endothelialization of "off-the-shelf," small-caliber arterial graft prostheses following implantation. In the present work, we evaluated the behavior of EOCs on thromboresistant graft coatings based on the collagen-mimetic protein Scl2-2 and poly(ethylene glycol) (PEG) diacrylate. Specifically, the attachment, proliferation, migration, and phenotype of EOCs on PEG-Scl2-2 hydrogels were evaluated as a function of Scl2-2 concentration (4, 8, and 12 mg/mL) relative to human umbilical vein endothelial cells (HUVECs). Results demonstrate the ability of each PEG-Scl2-2 hydrogel formulation to support EOC and HUVEC adhesion, proliferation, and spreading. However, only the 8 and 12 mg/mL PEG-Scl2-2 hydrogels were able to support stable EOC and HUVEC confluence. These PEG-Scl2-2 formulations were, therefore, selected for evaluation of their impact on EOC and HUVEC phenotype relative to PEG-collagen hydrogels. Cumulatively, both gene and protein level data indicated that 8 mg/mL PEG-Scl2-2 hydrogels supported similar or improved levels of EOC maturation relative to PEG-collagen controls based on evaluation of CD34, VEGFR2, PECAM-1, and VE-Cadherin. The 8 mg/mL PEG-Scl2-2 hydrogels also appeared to support similar or improved levels of EOC homeostatic marker expression relative to PEG-collagen hydrogels based on von Willebrand factor, collagen IV, NOS3, thrombomodulin, and E-selectin assessment. Combined, the present results indicate that PEG-Scl2-2 hydrogels warrant further investigation as "off-the-shelf" graft coatings. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1712-1724, 2017.


[Statistical analysis of maxillary sinus squamous cell carcinoma].
Cases of squamous cell carcinoma of the maxillary sinus initially treated at Keio University Hospital between January 1981 and December 1998 studied retrospectively involved 60 untreated cases--46 men and 14 women aged 36 to 86 years (mean: 59.8 years). Of these, 7 had T2, 41 T3 and 12 T4 tumors based on 1997 AJCC TNM classification. Seven (11.7%) had nodal involvement but none had distant metastasis at diagnosis. Of the 60, 53 (88.3%) were stage III or IV. Prognostic factors and survival were statistically analyzed for 59 cases, excluding the 60th who died of another cause. Follow-up was 4 to 227 months, with a median of 59 months and a mean of 38 months. Of the 59, 48 (81.4%) underwent neoadjuvant chemotherapy (NAC). Survival was estimated using the Kaplan-Meier method as univariate analysis. Cause-specific 5-year survival was 56.8% for all stages, 68.2% for T2, 60.0% for T3, and 41.7% for T4. T stage classification was thus a significant independent prognostic factor in multivariate analysis using Cox's proportional hazards model (p = 0.0240). It also identified T stage classification (p = 0.0486) and NAC (p = 0.0419) as significant independent factors affecting survival with organ preservation. We also statistically analyzed 48 cases treated with NAC, which showed complete response (CR) for 11, partial response (PR) for 25, no change (NC) for 11, and progressive disease (PD) for 1. Responders (CR + PR) showed significantly better survival and organ preservation than nonresponders (NC + PD). The NAC response enables us to predict prognosis. T4 cases without NAC response should be treated intensively.


Association of Prehospital Time to In-Hospital Trauma Mortality in a Physician-Staffed Emergency Medicine System.
The association between total prehospital time and mortality in physician-staffed trauma systems remains uncertain. To describe the association of total prehospital time and in-hospital mortality in prehospital, physician-staffed trauma systems in France, with the hypothesis that total prehospital time is associated with increased mortality. This cohort study was conducted from January 2009 to December 2016. Data for this study were derived from 2 distinct regional trauma registries in France (1 urban and 1 rural) that both have a physician-staffed emergency medical service. Consecutive adult trauma patients admitted to either of the regional trauma referral centers during the study period were included. Data analysis took place from March 2018 to September 2018. The association between death and prehospital time was assessed with a multivariable model adjusted with confounders. Total prehospital time was the primary exposure variable, recorded as the time from the arrival of the physician-led prehospital care team on scene to the arrival at the hospital. The main outcome of interest was all-cause in-hospital mortality. A total of 10 216 patients were included (mean [SD] age, 41 [18] years; 7937 men [78.3%]) affected by predominantly nonpenetrating injuries (9265 [91.5%]), with a mean (SD) Injury Severity Score of 17 (14) points. Of the patients, 6737 (66.5%) had at least 1 body region with an Abbreviated Injury Scale score of 3 or more. A total of 1259 patients (12.4%) presented in shock (with systolic pressure <90 mm Hg) and 2724 (26.9%) with severe head injury (Abbreviated Injury Scale score ≥3 points). On unadjusted analysis, increasing prehospital times (in 30-minute categories) were associated with a markedly and constant increase in the risk of in-hospital death. The odds of death increased by 9% for each 10-minute increase in prehospital time (odds ratio, 1.09 [95% CI, 1.07-1.11]) and after adjustment by 4% (odds ratio, 1.04 [95% CI, 1.01-1.07]). In this study, an increase in total prehospital time was associated with increasing in-hospital all-cause mortality in trauma patients at a physician-staffed emergency medical system, after adjustment for case complexity. Prehospital time is a management objective in analogy to physiological targets. These findings plead for a further streamlining of prehospital trauma care and the need to define the optimal intervention-to-time ratio.


Physician knowledge and practices in the evaluation of coagulopathies in stroke patients.
Coagulopathies are a rare cause of ischemic stroke. Prior studies demonstrate that current physician test-ordering practices for the evaluation of these conditions in patients with ischemic stroke is not optimal. We sought to determine neurologists' views regarding their use of specialized coagulation testing to better understand the possible reasons for these practices. A survey with multiple-choice and open-ended questions regarding knowledge of and approaches to the evaluation of coagulopathies was sent to a convenience sample of 79 neurologists (26 academic neurology faculty, 24 residents/fellows, and 29 community-based practitioners). Fifty-nine (75%) surveys were completed (response rates: faculty 73%, residents/fellows 88%, and community-based practice 66%). Specialized coagulation tests were reported to infrequently influence stroke patient management (<25% of the time or never for 95% of respondents). Factors reported to increase test-ordering included young patient age (76%), history of thrombosis (46%), history of miscarriages (36%), and having few traditional stroke risk factors (35%). Most (88%) indicated they would order specialized coagulation tests for a hypothetical young patient with no known stroke risk factors. In contrast, only 14% would obtain the tests for a patient having traditional stroke risk factors, and none would order the tests for a stroke patient with atrial fibrillation. Physician-reported practices for obtaining specialized coagulation tests differ from those found in observational studies in which more indiscriminate test ordering was observed. Closing knowledge gaps and improving application of physician's current knowledge to their test-ordering practices could help to optimize diagnostic testing for coagulopathies in patients with ischemic stroke.


[Ethical aspects of biomedical research. frequent concepts in written norms].
Most codes and declarations pertaining to the ethics of research involving human subjects have been formulated in response to specific events mostly of a painful or embarrassing nature. Several concepts appearing repeatedly in their texts are highlighted in this paper: principles of respect for persons, beneficence and justice, vulnerability and research propriety. Along with some comments on current revisions of many international guidelines and the role of bioethics committees, a plea is made to have a proactive rather than a reactive stance in ethical regulation of biomedical research.


Stressful life events and schizophrenia. II: Conceptual and methodological issues.
Research on the relationship between stress and schizophrenia is fraught with conceptual and methodological problems. These problems include issues related to the nature and measurement of stress, the likelihood of reciprocal influences between stress and symptoms, and the adequate assessment of symptoms. Several recommendations are made regarding future research in this area. These include using multiple and broadly based measures of different types of stressors and symptoms, greater use of truly prospective research designs, and the evaluation of the effects of interventions specifically designed to reduce stress in patients who suffer from schizophrenia.