Signal-to-noise contribution of principal component loads in reconstructed near-infrared Raman tissue spectra.
The overall quality of Raman spectra in the near-infrared region, where biological samples are often studied, has benefited from various improvements to optical instrumentation over the past decade. However, obtaining ample spectral quality for analysis is still challenging due to device requirements and short integration times required for (in vivo) clinical applications of Raman spectroscopy. Multivariate analytical methods, such as principal component analysis (PCA) and linear discriminant analysis (LDA), are routinely applied to Raman spectral datasets to develop classification models. Data compression is necessary prior to discriminant analysis to prevent or decrease the degree of over-fitting. The logical threshold for the selection of principal components (PCs) to be used in discriminant analysis is likely to be at a point before the PCs begin to introduce equivalent signal and noise and, hence, include no additional value. Assessment of the signal-to-noise ratio (SNR) at a certain peak or over a specific spectral region will depend on the sample measured. Therefore, the mean SNR over the whole spectral region (SNR(msr)) is determined in the original spectrum as well as for spectra reconstructed from an increasing number of principal components. This paper introduces a method of assessing the influence of signal and noise from individual PC loads and indicates a method of selection of PCs for LDA. To evaluate this method, two data sets with different SNRs were used. The sets were obtained with the same Raman system and the same measurement parameters on bladder tissue collected during white light cystoscopy (set A) and fluorescence-guided cystoscopy (set B). This method shows that the mean SNR over the spectral range in the original Raman spectra of these two data sets is related to the signal and noise contribution of principal component loads. The difference in mean SNR over the spectral range can also be appreciated since fewer principal components can reliably be used in the low SNR data set (set B) compared to the high SNR data set (set A). Despite the fact that no definitive threshold could be found, this method may help to determine the cutoff for the number of principal components used in discriminant analysis. Future analysis of a selection of spectral databases using this technique will allow optimum thresholds to be selected for different applications and spectral data quality levels.


Atopic dermatitis epidemiology and unmet need in the United Kingdom.
Atopic dermatitis (AD), also known as atopic eczema, is a chronic inflammatory skin condition associated with a significant health-related and socioeconomic burden, and is characterized by intense itch, disruption of the skin barrier, and upregulation of type 2-mediated immune responses. The United Kingdom (UK) has a high prevalence of AD, affecting 11-20% of children and 5-10% of adults. Approximately 2% of all cases of childhood AD in the UK are severe. Despite this, most AD treatments are performed at home, with little contact with healthcare providers or services. Here, we discuss the course of AD, treatment practices, and unmet need in the UK. Although the underlying etiology of the disease is still emerging, AD is currently attributed to skin barrier dysfunction and altered inflammatory responses. Management of AD focuses on avoiding triggers, improving skin hydration, managing exacerbating factors, and reducing inflammation through topical and systemic immunosuppressants. However, there is a significant unmet need to improve the overall management of AD and help patients gain control of their disease through safe and effective treatments. Approaches that target individual inflammatory pathways (e.g. dupilumab, anti-interleukin (IL)-4 receptor α) are emerging and likely to provide further therapeutic opportunities for patient benefit.


Changes in regional brain monoaminergic activity and temporary down-regulation in stress response from dietary supplementation with l-tryptophan in Atlantic cod (Gadus morhua).
The brain monoamines serotonin (5-hydroxytryptamine; 5-HT) and dopamine (DA) both play an integrative role in behavioural and neuroendocrine responses to challenges, and comparative models suggest common mechanisms for dietary modulation of transmission by these signal substances in vertebrates. Previous studies in teleosts demonstrate that 7 d of dietary administration with L-tryptophan (Trp), the direct precursor of 5-HT, suppresses the endocrine stress response. The present study investigated how long the suppressive effects of a Trp-enriched feed regimen, at doses corresponding to two, three or four times the Trp levels in commercial feed, last in juvenile Atlantic cod (Gadus morhua) when the fish are reintroduced to a diet with standard amino acid composition. We also wanted to determine whether Trp supplementation induced changes in brain monoaminergic neurochemistry in those forebrain structures innervated by DA and 5-HTergic neurons, by measuring regional activity of DA and 5-HT in the lateral pallial regions (Dl) of the telencephalon and nucleus lateralis tuberis (NLT) of the hypothalamus. Dietary Trp resulted in a dose-dependent suppression in plasma cortisol among fish exposed to confinement stress on the first day following experimental diet; however, such an effect was not observed at 2 or 6 d after Trp treatment. Feeding the fish with moderate Trp doses also evoked a general increase in DA and 5-HT-ergic activity, suggesting that these neural circuits within the NLT and Dl may be indirectly involved in regulating the acute stress response.


Seasonal responses of xylem sap velocity to VPD and solar radiation during drought in a stand of native trees in temperate Australia.
Xylem sap velocity of two dominant tree species, Eucalyptus crebra F. Muell. and Callitris glaucophylla J. Thompson & L.A.S. Johnson, in a native remnant forest of eastern Australia was measured in winter and summer during a prolonged (> 12 months) and extensive drought. The influence of vapour pressure deficit (VPD) and solar radiation levels on the velocity of sap was determined. Pronounced hysteresis in sap velocity was observed in both species as a function of VPD and solar radiation. However, the rotation of the hysteresis curve was clockwise for the response of sap velocity to VPD but anti-clockwise in the response of sap velocity to radiation levels. A possible reason for this difference is discussed.The degree of hysteresis (area bounded by the curve) was larger for the VPD response than the response to solar radiation and also varied with season. A simple linear model was able to predict sap velocity from knowledge of VPD and solar radiation in winter and summer. The consistent presence of hysteresis in the response to sap velocity to VPD and solar radiation suggests that large temporal and spatial models of vegetation water use may require some provision for the different responses of sap velocity, and hence water use, to VPD and solar radiation, between morning and afternoon and between seasons.


Non-Surgical Retreatment After Failed Intentional Replantation: A Case Report.
A 27-year-old male patient complained of dental pain on previously root-treated tooth #17. Intentional replantation (IR) was performed. However, the symptoms returned at 10 months of follow-up, and a sinus tract was detected on the buccal gingiva of tooth #17. The patient refused extraction of the tooth and tooth was scheduled for non-surgical retreatment. Root canal retreatment was initiated with the removal of the previous root filling material while leaving the retrograde filling in situ. After 1 month of intracanal medication, the tooth became asymptomatic with complete healing of the sinus tract and the root canal filling was completed. Clinical and radiographic examination at 1 year follow-up revealed endodontic success with complete resolution of periapical radiolucency. This case report shows that non-surgical retreatment could be a viable alternative for failed IR.


Mongersen, an oral SMAD7 antisense oligonucleotide, and Crohn's disease.
Crohn's disease-related inflammation is characterized by reduced activity of the immunosuppressive cytokine transforming growth factor β1 (TGF-β1) due to high levels of SMAD7, an inhibitor of TGF-β1 signaling. Preclinical studies and a phase 1 study have shown that an oral SMAD7 antisense oligonucleotide, mongersen, targets ileal and colonic SMAD7. In a double-blind, placebo-controlled, phase 2 trial, we evaluated the efficacy of mongersen for the treatment of persons with active Crohn's disease. Patients were randomly assigned to receive 10, 40, or 160 mg of mongersen or placebo per day for 2 weeks. The primary outcomes were clinical remission at day 15, defined as a Crohn's Disease Activity Index (CDAI) score of less than 150, with maintenance of remission for at least 2 weeks, and the safety of mongersen treatment. A secondary outcome was clinical response (defined as a reduction of 100 points or more in the CDAI score) at day 28. The proportions of patients who reached the primary end point were 55% and 65% for the 40-mg and 160-mg mongersen groups, respectively, as compared with 10% for the placebo group (P<0.001). There was no significant difference in the percentage of participants reaching clinical remission between the 10-mg group (12%) and the placebo group. The rate of clinical response was significantly greater among patients receiving 10 mg (37%), 40 mg (58%), or 160 mg (72%) of mongersen than among those receiving placebo (17%) (P=0.04, P<0.001, and P<0.001, respectively). Most adverse events were related to complications and symptoms of Crohn's disease. We found that study participants with Crohn's disease who received mongersen had significantly higher rates of remission and clinical response than those who received placebo. (Funded by Giuliani; EudraCT number, 2011-002640-27.).


High-conductive organometallic molecular wires with delocalized electron systems strongly coupled to metal electrodes.
Besides active, functional molecular building blocks such as diodes or switches, passive components, for example, molecular wires, are required to realize molecular-scale electronics. Incorporating metal centers in the molecular backbone enables the molecular energy levels to be tuned in respect to the Fermi energy of the electrodes. Furthermore, by using more than one metal center and sp-bridging ligands, a strongly delocalized electron system is formed between these metallic "dopants", facilitating transport along the molecular backbone. Here, we study the influence of molecule-metal coupling on charge transport of dinuclear X(PP)2FeC4Fe(PP)2X molecular wires (PP = Et2PCH2CH2PEt2); X = CN (1), NCS (2), NCSe (3), C4SnMe3 (4), and C2SnMe3 (5) under ultrahigh vacuum and variable temperature conditions. In contrast to 1, which showed unstable junctions at very low conductance (8.1 × 10(-7) G0), 4 formed a Au-C4FeC4FeC4-Au junction 4' after SnMe3 extrusion, which revealed a conductance of 8.9 × 10(-3) G0, 3 orders of magnitude higher than for 2 (7.9 × 10(-6) G0) and 2 orders of magnitude higher than for 3 (3.8 × 10(-4) G0). Density functional theory (DFT) confirmed the experimental trend in the conductance for the various anchoring motifs. The strong hybridization of molecular and metal states found in the C-Au coupling case enables the delocalized electronic system of the organometallic Fe2 backbone to be extended over the molecule-metal interfaces to the metal electrodes to establish high-conductive molecular wires.


Identifying loci for the overlap between attention-deficit/hyperactivity disorder and autism spectrum disorder using a genome-wide QTL linkage approach.
The genetic basis for autism spectrum disorder (ASD) symptoms in children with attention-deficit/hyperactivity disorder (ADHD) was addressed using a genome-wide linkage approach. Participants of the International Multi-Center ADHD Genetics study comprising 1,143 probands with ADHD and 1,453 siblings were analyzed. The total and subscale scores of the Social Communication Questionnaire (SCQ) were used as quantitative traits for multipoint regression-based linkage analyses on 5,407 autosomal single-nucleotide polymorphisms applying MERLIN-regress software, both without and with inclusion of ADHD symptom scores as covariates. The analyses without ADHD symptom scores as covariates resulted in three suggestive linkage signals, i.e., on chromosomes 15q24, 16p13, and 18p11. Inclusion of ADHD symptom scores as covariates resulted in additional suggestive loci on chromosomes 7q36 and 12q24, whereas the LOD score of the locus on chromosome 15q decreased below the threshold for suggestive linkage. The loci on 7q, 16p, and 18p were found for the SCQ restricted and repetitive subscale, that on 15q was found for the SCQ communication subscale, and that on 12q for the SCQ total score. Our findings suggest that QTLs identified in this study are ASD specific, although the 15q QTL potentially has pleiotropic effects for ADHD and ASD. This study confirms that genetic factors influence ASD traits along a continuum of severity, as loci potentially underlying ASD symptoms in children with ADHD were identified even though subjects with autism had been excluded from the IMAGE sample, and supports the hypothesis that differential genetic factors underlie the three ASD dimensions.


Fluoride-induced lesions in the teeth of the short-tailed field vole (Microtus agrestis): a description of the dental pathology.
The effect of fluoride on the appearance of the teeth of the short-tailed field vole, Microtus agrestis, was investigated in both wild animals collected from field sites affected by different levels of industrial fluoride contamination and laboratory-reared animals consuming experimental grass diets of known fluoride concentration or with known fluoride concentrations in drinking water. The extent and severity of lesions on the surface and structure of both incisors and molars are described as six lesion types and related to the amount of biologically available orally ingested fluoride. In the incisors of voles consuming relatively low fluoride diets, lesions are mainly confined to those resulting from disruption of enamel pigmentation expressing itself as concentric bands of pigmentation-free areas on incisor surfaces. The visible effects on molars at low fluoride levels are confined to minor alterations in surface appearance. At higher levels of available dietary fluoride, effects on enamel pigmentation are superseded by alterations in the formation, composition, and strength of both enamel and dentine. The incisors exhibit a marked to severe increase in the cutting tip erosion rates with comparable increases in the extent of abnormal surface changes (enamel hypoplasia) and the loss of enamel pigmentation. The grinding surfaces of molars from animals exposed to high levels of dietary fluoride exhibit increasingly severe erosion of outer enamel regions, combined with cavitation and staining of the exposed central dentine. The mechanisms through which fluoride elicits increasingly visible and pathological alterations to the surface and subsurfaces of rodent teeth are discussed.


Thermally active TRPV1 tonically drives central spontaneous glutamate release.
Central synapses spontaneously release neurotransmitter at low rates. In the brainstem, cranial visceral afferent terminals in caudal solitary tract nucleus (NTS) display pronounced, activity-dependent, asynchronous release of glutamate and this extra release depends on TRPV1 receptors (TRPV1+). Asynchronous release is absent for afferents lacking TRPV1 (TRPV1-) and resting EPSC frequency was greater in TRPV1+. Here, we studied this basal activity difference by assessing thermal sensitivity of spontaneous and miniature synaptic events in TRPV1+ and TRPV1- second-order NTS neurons. The spontaneous EPSC rate decreased when temperature was decreased, increased steeply between 30 and 42°C only in TRPV1+ neurons, and was calcium-dependent. TRPV1-specific antagonist SB366791, but not TTX, strongly attenuated thermal responses. Temperature changes failed to alter EPSC frequency in TRPV1- neurons. EPSC amplitudes and decay kinetics changed little with temperature. IPSCs in these second-order NTS neurons were unaltered by temperature. Such results suggest that activated, presynaptic TRPV1+ receptors trigger continuous resting release of glutamate vesicles at physiological temperatures only in capsaicin-responsive terminals. In mechanically isolated individual neurons harvested from medial NTS, increases in temperature increased the rate of glutamate release only in TRPV1+ neurons, whereas IPSC rates were unaffected. Cadmium failed to block thermal increases in glutamate release, suggesting that calcium entry through TRPV1 channels may trigger glutamate release independently of voltage-activated calcium channels. Together, our findings indicate a new form of afferent signaling in which TRPV1 channels within central terminals of peripheral afferents tonically generate glutamate release in NTS at 37°C in the absence of afferent action potentials.


Seed dormancy-breaking in a cold desert shrub in relation to sand temperature and moisture.
Seasonal periodicity of seed germination and its relationship to seasonal changes in temperature and soil moisture have been well studied in seeds of species with physiological dormancy. However, relatively little information is available on the role of these environmental factors in controlling germination of seeds with physical dormancy (PY). Our primary aim was to determine if seeds of the cold desert sand dune semi-shrub Eremosparton songoricum exhibits seasonal periodicity of seed germination and the relationship between seed dormancy break and soil temperature and moisture. In the laboratory, seeds incubated on dry, wet, wet-dry and dry-wet sand were exposed to a 1-year sequence of temperature regimes simulating those in the field. In the field, seeds were buried at different depths on a sand dune, and germination of periodically exhumed seeds was tested at five temperature regimes during a 2-year period. In the one-year sequence of simulated natural temperature regimes, breaking of PY was more effective under constantly wet than under constantly dry conditions, and germination percentage was significantly higher under dry-wet than under wet-dry conditions. Seeds buried in the field exhibited a distinct peak of germination in spring and little or no germination in other seasons. The final (two-year) monthly cumulative germination percentage differed among burial depths and temperature, and it was highest (47%) in seeds buried at 3 cm and tested at 25/10 °C. A seed cohort of E. songoricum likely exhibits a long-term annual periodicity of spring germination in the field, and dormancy break appears to be driven by low (winter) temperatures and relatively high sand moisture content. To our knowledge this is the first study to document seasonal periodicity in seed germination in a cold desert species with PY and to identify the mechanism (at the whole-seed level) of its occurrence.


Granulocyte-colony stimulating factor improves outcome in a mouse model of amyotrophic lateral sclerosis.
Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disease that results in progressive loss of motoneurons, motor weakness and death within 1-5 years after disease onset. Therapeutic options remain limited despite a substantial number of approaches that have been tested clinically. In particular, various neurotrophic factors have been investigated. Failure in these trials has been largely ascribed to problems of insufficient dosing or inability to cross the blood-brain barrier (BBB). We have recently uncovered the neurotrophic properties of the haematopoietic protein granulocyte-colony stimulating factor (G-CSF). The protein is clinically well tolerated and crosses the intact BBB. This study examined the potential role of G-CSF in motoneuron diseases. We investigated the expression of the G-CSF receptor in motoneurons and studied effects of G-CSF in a motoneuron cell line and in the SOD1(G93A) transgenic mouse model. The neurotrophic growth factor was applied both by continuous subcutaneous delivery and CNS-targeted transgenic overexpression. This study shows that given at the stage of the disease where muscle denervation is already evident, G-CSF leads to significant improvement in motor performance, delays the onset of severe motor impairment and prolongs overall survival of SOD1(G93A)tg mice. The G-CSF receptor is expressed by motoneurons and G-CSF protects cultured motoneuronal cells from apoptosis. In ALS mice, G-CSF increased survival of motoneurons and decreased muscular denervation atrophy. We conclude that G-CSF is a novel neurotrophic factor for motoneurons that is an attractive and feasible drug candidate for the treatment of ALS.


Molecular characterization of non-subtype C and recombinant HIV-1 viruses from Cape Town, South Africa.
HIV was first diagnosed within South Africa in 1982. Homosexual transmission of HIV-1 dominated the epidemic within the country in the early stages of the 1980s. Currently heterosexual transmission of HIV-1 is responsible for the majority of HIV cases in South Africa with subtype C HIV-1 being responsible for an estimated 95% of infections. Only a few papers have been published on non-subtype C HIV-1 detection within South Africa. This study characterized subgenomic and near full-length sequences of non-subtype C HIV-1 viruses from the Cape Town area. Amplification and direct sequencing characterized partial gene fragments of 11 samples. Phylogenetic analysis of the sequenced data, with online subtyping tools (REGA and jpHMM) and the drawing of NJ-trees revealed the presence of subtypes A1, B, and F1 as well as recombinant viral forms such as AD, AG and AC. Near full-length genome characterization of 4 of the 11 samples was performed. Analysis of sequenced data with the use of subtyping, recombination identification, and tree drawing tools revealed one subtype B and one A1 isolate. The other two isolates were identified as AC and AD recombinants. The data that was gathered will greatly improve our knowledge of non-subtype C isolates circulating within South Africa.


Market-Based Health Care in Specialty Surgery: Finding Patient-Centered Shared Value.
: The US health care system is struggling with rising costs, poor outcomes, waste, and inefficiency. The Patient Protection and Affordable Care Act represents a substantial effort to improve access and emphasizes value-based care. Value in health care has been defined as health outcomes for the patient per dollar spent. However, given the opacity of health outcomes and cost, the identification and quantification of patient-centered value is problematic. These problems are magnified by highly technical, specialized care (eg, neurosurgery). This is further complicated by potentially competing interests of the 5 major stakeholders in health care: patients, doctors, payers, hospitals, and manufacturers. These stakeholders are watching with great interest as health care in the United States moves toward a value-based system. Market principles can be harnessed to drive costs down, improve outcomes, and improve overall value to patients. However, there are many caveats to a market-based, value-driven system that must be identified and addressed. Many excellent neurosurgical efforts are already underway to nudge health care toward increased efficiency, decreased costs, and improved quality. Patient-centered shared value can provide a philosophical mooring for the development of health care policies that utilize market principles without losing sight of the ultimate goals of health care, to care for patients.


Causes and consequences of sea urchin abundance and diversity in Kenyan coral reef lagoons.
Large differences in community structure of sea urchins and finfish have been observed in Kenyan reef lagoons. Differences have been attributed to removal of finfish predators through human fishing activities. This study attempted to determine (i) the major sea urchin finfish predators, (ii) the effect of predation on sea-urchin community structure, and (iii) the possible effect of sea urchin increases and finfish decreases on the lagoonal substrate. Six reefs, two protected and four unprotected, were compared for differences in finfish abundance, sea urchin abundance and diversity and substrate cover, diversity and complexity. Comparisons between protected and unprotected reefs indicated that finfish populations were ca. 4 x denser in protected than unprotected reefs. Sea urchin populations were >100 x denser and predation rates on a sea urchin, Echinometra mathaei, were 4 x lower in unprotected than in protected reefs. The balistidae (triggerfish) was the single sea-urchin finfish predator family which had a higher population density in protected than in unprotected reefs. Balistid density was positively correlated with predation rates on tethered E. mathaei (r=0.88; p<0.025) and negatively correlated with total sea-urchin density (r=-0.89; p<0.025) on the six reefs. We conclude from observations that the balistids Balistaphus undulatus and Rhinecanthus aculeatus are the dominant sea-urchin predators. The sea-urchin assemblage had its greatest diversity and species richness at intermediate predation rates and low to intermediate sea-urchin densities. At low predation rates and high sea-urchin density E. mathaei dominated the assemblage's species composition. Preferential predation on the competitive dominant maintains the assemblage's diversity, supporting the compensatory mortality hypothesis (Connell 1978) of coral reef diversity. Protected reefs had greater cover of hard coral, calcareous and coralline algae, and greater substrate diversity and topographic complexity than unprotected reefs which had greater algal turf and sponge cover. Coral cover and topographic complexity were negatively correlated with total sea urchin density. Although experimentation is lacking, these substrate changes may be due to the switch from finfish to sea-urchins as consumers which results from overfishing of finfish. Removal of top invertebrate-eating carnivores appears to have cascading effects on the entire coral reef ecosystem.


Serotonin transporter [corrected] methylation and response to cognitive behaviour therapy in children with anxiety disorders.
Anxiety disorders that are the most commonly occurring psychiatric disorders in childhood, are associated with a range of social and educational impairments and often continue into adulthood. Cognitive behaviour therapy (CBT) is an effective treatment option for the majority of cases, although up to 35-45% of children do not achieve remission. Recent research suggests that some genetic variants may be associated with a more beneficial response to psychological therapy. Epigenetic mechanisms such as DNA methylation work at the interface between genetic and environmental influences. Furthermore, epigenetic alterations at the serotonin transporter (SERT) promoter region have been associated with environmental influences such as stressful life experiences. In this study, we measured DNA methylation upstream of SERT in 116 children with an anxiety disorder, before and after receiving CBT. Change during treatment in percentage DNA methylation was significantly different in treatment responders vs nonresponders. This effect was driven by one CpG site in particular, at which responders increased in methylation, whereas nonresponders showed a decrease in DNA methylation. This is the first study to demonstrate differences in SERT methylation change in association with response to a purely psychological therapy. These findings confirm that biological changes occur alongside changes in symptomatology following a psychological therapy such as CBT.


Mobile Rayleigh Doppler lidar for wind and temperature measurements in the stratosphere and lower mesosphere.
A mobile Rayleigh Doppler lidar based on the molecular double-edge technique is developed for measuring wind velocity in the middle atmosphere up to 60 km. The lidar uses three lasers with a mean power of 17.5 W at 355 nm each and three 1 m diameter telescopes to receive the backscattered echo: one points to zenith for vertical wind component and temperature measurement; the two others pointing toward east and north are titled at 30° from the zenith for zonal and meridional wind component, respectively. The Doppler shift of the backscattered echo is measured by inter-comparing the signal detected through each of the double-edge channels of a triple Fabry-Perot interferometer (FPI) tuned to either side of the emitted laser line. The third channel of FPI is used for frequency locking and a locking accuracy of 1.8 MHz RMS (root-mean-square) at 355 nm over 2 hours is realized, corresponding to a systematic error of 0.32 m/s. In this paper, we present detailed technical evolutions on system calibration. To validate the performance of the lidar, comparison experiments was carried out in December 2013, which showed good agreement with radiosondes but notable biases with ECMWF (European Centre for Medium range Weather Forecasts) in the height range of overlapping data. Wind observation over one month performed in Delhi (37.371° N, 97.374° E), northwest of China, demonstrated the stability and robustness of the system.


DNA sequence-dependent activity and base flipping mechanisms of DNMT1 regulate genome-wide DNA methylation.
DNA methylation maintenance by DNMT1 is an essential process in mammals but molecular mechanisms connecting DNA methylation patterns and enzyme activity remain elusive. Here, we systematically analyzed the specificity of DNMT1, revealing a pronounced influence of the DNA sequences flanking the target CpG site on DNMT1 activity. We determined DNMT1 structures in complex with preferred DNA substrates revealing that DNMT1 employs flanking sequence-dependent base flipping mechanisms, with large structural rearrangements of the DNA correlating with low catalytic activity. Moreover, flanking sequences influence the conformational dynamics of the active site and cofactor binding pocket. Importantly, we show that the flanking sequence preferences of DNMT1 highly correlate with genomic methylation in human and mouse cells, and 5-azacytidine triggered DNA demethylation is more pronounced at CpG sites with flanks disfavored by DNMT1. Overall, our findings uncover the intricate interplay between CpG-flanking sequence, DNMT1-mediated base flipping and the dynamic landscape of DNA methylation.


Detection of a milk factor that facilitates folate uptake by intestinal cells.
Folate binding proteins in milk were tested for their effect on folate absorption. The uptake of bound folate by isolated mucosal cells from the rat small intestine was twice that of free folate and differed from it in being more effective with progression down the small intestine, in not being affected by glucose or Dilantin, in having a higher pH optimum, and in being affected by calcium concentration. This milk factor may enhance folate absorption in infants, whose risk of folate deficiency is high.


Implementation Adherence and Perspectives of the Childcare PhysicaL ActivitY (PLAY) Policy: A Process Evaluation.
The Childcare PhysicaL ActivitY (PLAY) policy was an evidence-informed, eight-item institutional-level policy document targeting children's physical activity, outdoor play, and sedentary time. Nine childcare centers in London, Ontario, participated in this cluster, randomized controlled trial. Early Childhood Educators allocated to the experimental group, from five childcare centers in London, Ontario, implemented the policy for young children (18 months to 4 years) for 8 weeks and documented adherence to each policy item (i.e., dose) in daily logs. Program evaluation surveys (n = 21) and interviews (n = 10) were completed postintervention to assess Early Childhood Educators' perspectives of feasibility, context, enjoyment, communication between researchers and childcare staff, and likelihood of future implementation. Descriptive statistics were calculated, and thematic analysis was conducted. Adherence to policy items ranged from 16.5% (for delivery of shorter, more frequent outdoor periods) to 85.9% (for delivery of unstructured/child-directed play). Participants reported effective communication between the research team and childcare centers (0 = not at all effective to 5 = very effective; M = 4.20; SD = 0.83) but noted that they were unlikely to continue the implementation of more frequent outdoor periods (0 = not at all likely to 5 = extremely likely; M = 2.19; SD = 1.21). Interview themes included weather as a prominent barrier and the use of verbal prompts as a solution for implementing the policy. As this was a small and short-term intervention, this pilot study offers important insight on larger scale policy interventions aimed at increasing physical activity and minimizing sedentary time among children enrolled in childcare.


Establishing a Scientific Consensus on the Cognitive Benefits of Physical Activity.
Research suggests that physical activity can be used as an intervention to increase cognitive function. Yet, there are competing views on the cognitive effects of physical activity and it is not clear what level of consensus exists among researchers in the field. The purpose of this study was two-fold: Firstly, to quantify the scientific consensus by focusing on the relationship between physical activity and cognitive function. Secondly, to investigate if there is a gap between the public's and scientists' interpretations of scientific texts on this topic. A two-phase study was performed by including 75 scientists in the first phase and 15 non-scientists in the second phase. Participants were asked to categorize article abstracts in terms of endorsement of the effect of physical activity on cognitive function. Results indicated that there was a 76.1% consensus that physical activity has positive cognitive effects. There was a consistent association between scientists' and non-scientists' categorizations, suggesting that both groups perceived abstracts in a similar fashion. Taken together, this study provides the first analysis of its kind to evaluate the level of consensus in almost two decades of research. The present data can be used to inform further research and practice.


Epithelial localization of a reptilian Na+/H+ exchanger homologous to NHE-1.
Basolateral membranes of turtle (Pseudemys scripta) colon epithelial cells exhibit robust Na+/H+ exchange activity that can be activated by cell shrinkage and is blocked by amiloride [M. A. Post and D. C. Dawson. Am. J. Physiol. 262 Cell Physiol. 31):C1089-C1094, 1992]. The colonic epithelium actively absorbs Na+ and secretes K+ and HCO3-, but the role of basolateral Na+/H+ exchange, if any, in transepithelial transport is unknown. The current studies were undertaken to identify the gene product(s) responsible for the observed basolateral Na+/H+ exchange activity and to determine the cellular localization of the reptilian Na+/H+ exchange protein. We cloned and sequenced partial-length cDNAs that are likely to encode a reptilian homologue of the mammalian NHE-1 Na+/H+ exchanger isoform. The partial-length cDNAs were > 80% identical to mammalian NHE-1 homologues at the nucleotide level and recognized a transcript (approximately 5.8-6.0 kb) in RNA isolated from turtle colon, small intestine, stomach, kidney, urinary bladder, heart, and liver. In situ hybridization showed that mRNA encoding the reptile homologue of NHE-1 was expressed predominantly in the epithelial cells of these tissues. Immunofluorescent localization of the reptilian Na+/H+ exchanger protein using an antibody raised against a human NHE-1 fusion protein confirmed that protein expression paralleled abundant mRNA expression in epithelial cells of turtle stomach and colon, as well as in some nephron segments, and showed that the reptile NHE-1 homologue was localized exclusively to the basolateral membranes of these cells. The relatively high level of NHE-1 expression in epithelial cells, particularly those of the colon and stomach, suggests that NHE-1 function is important for the maintenance or regulation of ion transport processes that occur in these cell types.


Syntrophy emerges spontaneously in complex metabolic systems.
Syntrophy allows a microbial community as a whole to survive in an environment, even though individual microbes cannot. The metabolic interdependence typical of syntrophy is thought to arise from the accumulation of degenerative mutations during the sustained co-evolution of initially self-sufficient organisms. An alternative and underexplored possibility is that syntrophy can emerge spontaneously in communities of organisms that did not co-evolve. Here, we study this de novo origin of syntrophy using experimentally validated computational techniques to predict an organism's viability from its metabolic reactions. We show that pairs of metabolisms that are randomly sampled from a large space of possible metabolism and viable on specific primary carbon sources often become viable on new carbon sources by exchanging metabolites. The same biochemical reactions that are required for viability on primary carbon sources also confer viability on novel carbon sources. Our observations highlight a new and important avenue for the emergence of metabolic adaptations and novel ecological interactions.


Self-esteem, metacognition, and coping strategies in cancer patients: A case-control study.
Self-esteem refers to a person's positive and negative attitudes towards the self, and metacognition is an upper system providing awareness and direction of events and mental functions. Coping refers to the specific and psychological efforts used to deal with stressful events or the negative effects of the agents of these. The aim of this study was to evaluate self-esteem, metacognition status and coping attitudes in patients with cancer, which is known to have severely destructive psychological effects. Fifty adult cancer patients who were followed up in the medical oncology clinic between July 2018 and June 2019 and 50 age- and gender-matched healthy controls as control group were included in this study. All the participants were applied with a sociodemographic data form, the Rosenberg self-esteem scale, the Metacognition Assessment Scale, and the Copying Orientation to Problems Experienced (COPE) inventory, and their results were compared between the groups. The groups comprised 50% females with a median age of 58 (33-82) years. The values related to the degree of participation in discussions, problem-focused coping, active coping, planning, and state of emotional vulnerability were low in the cancer patient group compared to the control group (P < 0.005 for all). The sustaining of their self-image, feeling threatened in interpersonal relationships, and degree of daydreaming were higher, and in the metacognition tests, the positive beliefs related to anxiety, uncontrolled or dangerous negative thoughts, nonfunctional coping, religious coping, joking, reckless behavior, substance use, denial, and mental disengagement scores were higher (P < 0.05 for all). Self-esteem was lower in cancer patients and upper level cognitive functions and problem-focused coping were determined to be worse compared to healthy controls. In the light of these results, psychosocial support given to cancer patients in this respect could contribute to quality of life and social conformity.


Alignment of cryo-electron tomography datasets.
Data acquisition of cryo-electron tomography (CET) samples described in previous chapters involves relatively imprecise mechanical motions: the tilt series has shifts, rotations, and several other distortions between projections. Alignment is the procedure of correcting for these effects in each image and requires the estimation of a projection model that describes how points from the sample in three-dimensions are projected to generate two-dimensional images. This estimation is enabled by finding corresponding common features between images. This chapter reviews several software packages that perform alignment and reconstruction tasks completely automatically (or with minimal user intervention) in two main scenarios: using gold fiducial markers as high contrast features or using relevant biological structures present in the image (marker-free). In particular, we emphasize the key decision points in the process that users should focus on in order to obtain high-resolution reconstructions.


Orientation priming of novel shapes in the context of viewpoint-dependent recognition.
Can visual similarity between shapes facilitate orientation priming? Five experiments are reported in which this possibility was explored by using novel two-dimensional shapes that formed homogeneous stimulus classes. After training on individual shapes in a canonical view, the recognition of these shapes was tested in several picture-plane orientations. In experiments 1 and 2 an identification task was used to replicate the classic finding obtained with the mirror-judgment task-that prior orientation cueing does not reduce the magnitude of orientation dependence in processing rotated shapes. The results of experiment 3, however, indicate that blocking trials by orientation is one condition in which orientation priming may be obtained. Experiment 4 builds on this result, and it is suggested that awareness of the blocking manipulation is not required to obtain orientation priming. In experiment 5 the mechanisms underlying this finding are explored, and evidence is offered that orientation priming is a consequence of representations that encode both shape and orientation. Such results may be considered as an extension to the 'image-based' approach to object recognition, demonstrating that generalization across exemplars may occur within recognition mechanisms that are viewpoint dependent.


Plasma from human mothers of fetuses with severe arthrogryposis multiplex congenita causes deformities in mice.
Arthrogryposis multiplex congenita (AMC) is characterized by fixed joint contractures and other deformities, sometimes resulting in fetal death. The cause is unknown in most cases, but some women with fetuses affected by severe AMC have serum antibodies that inhibit fetal acetylcholine receptor (AChR) function, and antibodies to fetal antigens might play a pathogenic role in other congenital disorders. To investigate this possibility, we have established a model by injecting pregnant mice with plasma from four anti-AChR antibody-positive women whose fetuses had severe AMC. We found that human antibodies can be transferred efficiently to the mouse fetus during the last few days of fetal life. Many of the fetuses of dams injected with AMC maternal plasmas or Ig were stillborn and showed fixed joints and other deformities. Moreover, similar changes were found in mice after injection of a serum from one anti-AChR antibody-negative mother who had had four AMC fetuses. Thus, we have confirmed the role of maternal antibodies in cases of AMC associated with maternal anti-AChR, and we have demonstrated the existence of pathogenic maternal factors in one other case. Importantly, this approach can be used to look at the effects of other maternal human antibodies on development of the fetus.


Cellular Stress and p53-Associated Apoptosis by Juniperus communis L. Berry Extract Treatment in the Human SH-SY5Y Neuroblastoma Cells.
Plant phenolics have shown to activate apoptotic cell death in different tumourigenic cell lines. In this study, we evaluated the effects of juniper berry extract (Juniperus communis L.) on p53 protein, gene expression and DNA fragmentation in human neuroblastoma SH-SY5Y cells. In addition, we analyzed the phenolic composition of the extract. We found that juniper berry extract activated cellular relocalization of p53 and DNA fragmentation-dependent cell death. Differentially expressed genes between treated and non-treated cells were evaluated with the cDNA-RDA (representational difference analysis) method at the early time point of apoptotic process when p53 started to be activated and no caspase activity was detected. Twenty one overexpressed genes related to cellular stress, protein synthesis, cell survival and death were detected. Interestingly, they included endoplasmic reticulum (ER) stress inducer and sensor HSPA5 and other ER stress-related genes CALM2 and YKT6 indicating that ER stress response was involved in juniper berry extract mediated cell death. In composition analysis, we identified and quantified low concentrations of fifteen phenolic compounds. The main groups of them were flavones, flavonols, phenolic acids, flavanol and biflavonoid including glycosides of quercetin, apigenin, isoscutellarein and hypolaetin. It is suggested that juniper berry extract induced the p53-associated apoptosis through the potentiation and synergism by several phenolic compounds.


Phase transformations in binary colloidal monolayers.
Phase transformations can be difficult to characterize at the microscopic level due to the inability to directly observe individual atomic motions. Model colloidal systems, by contrast, permit the direct observation of individual particle dynamics and of collective rearrangements, which allows for real-space characterization of phase transitions. Here, we study a quasi-two-dimensional, binary colloidal alloy that exhibits liquid-solid and solid-solid phase transitions, focusing on the kinetics of a diffusionless transformation between two crystal phases. Experiments are conducted on a monolayer of magnetic and nonmagnetic spheres suspended in a thin layer of ferrofluid and exposed to a tunable magnetic field. A theoretical model of hard spheres with point dipoles at their centers is used to guide the choice of experimental parameters and characterize the underlying materials physics. When the applied field is normal to the fluid layer, a checkerboard crystal forms; when the angle between the field and the normal is sufficiently large, a striped crystal assembles. As the field is slowly tilted away from the normal, we find that the transformation pathway between the two phases depends strongly on crystal orientation, field strength, and degree of confinement of the monolayer. In some cases, the pathway occurs by smooth magnetostrictive shear, while in others it involves the sudden formation of martensitic plates.


Loss of apical vertebral derotation in adolescent idiopathic scoliosis: 2-year follow-up using multi-planar reconstruction computed tomography.
The objective of this study was to evaluate 2 years post-surgical loss of three-dimensional correction in adolescent idiopathic scoliosis (AIS) patients using multi-planar reconstruction computed tomography (CT). Twenty-seven AIS patients treated by segmental pedicle screw (PS) constructs were included in this study. Correction in the axial plane was evaluated using the "relative apical vertebral rotation angle" (rAVR), defined as the difference between the axial rotation angles of the upper instrumented vertebra and the apical vertebra on reconstructed axial CT images. The Cobb angle of the main curve and apical vertebral translation was measured to evaluate the coronal correction. Thoracic kyphosis was also measured for the evaluation of sagittal profile. Measurements were performed before surgery, and 1 week and 2 years after surgery. The relationships between the correction losses and skeletal maturity, and variety of spinal constructs were also evaluated. The mean preoperative Cobb angle of the major curve was 59.1° ± 11.2° before and 13.0° ± 7.2° immediately after surgery. Two years later, the mean Cobb angle had increased significantly, to 15.5° ± 7.8°, with a mean correction loss of 2.5° ± 1.5° (p < 0.001). The mean preoperative rAVR of 28.5° ± 8.4° was corrected to 15.8° ± 7.8° after surgery. It had increased significantly to 18.5 ± 8.4 by 2 years after surgery, with a mean correction loss of 2.7° ± 1.0° (p < 0.001). The mean correction losses for both the Cobb angle and rAVR were significantly greater in the skeletally immature patients. The significant correlations were recognized between the correction losses and the proportion of multi-axial screws, and the materials of constructs. Statistically significant loss of correction in the Cobb angle and apical vertebral axial rotation angle (AVR) were recognized 2 years after surgery using PS constructs. The correction losses, especially AVR, were more evident in the skeletally immature patients, and in patients treated with more multi-axial screws and with titanium constructs rather than with stainless constructs.


Alveolar Ridge Conservation by Early Bone Formation After Tooth Extraction in Rabbits. A Histomorphological Study.
Alveolar ridge volume loss is an irreversible process. To prevent this physiological event, which typically result in significant local anatomical changes in both the horizontal and the vertical dimension, some strategies are indicated to minimize the loss of ridge volume that typically follows tooth extraction. The purpose of this study was to evaluate if three different bone grafts could promote new bone formation in the alveolar socket following tooth extraction for the alveolar ridge conservation. First mandibular molars of male adults rabbits were extracted and the extraction sockets were randomly treated with three different bone grafts, one xenograft and two alloplastic grafts, and a group that received no treatment (blood clot). The extraction sockets of selected rabbits from each group were evaluated at 4, 6, or 8-week post-extraction. The results indicated that the extraction sockets treated with alloplastic graft (biphasic calcium phosphate) exhibited lamellar bone formation (6.5%) as early as four weeks after the extraction was performed. Moreover, the degree of new bone formation was significantly higher (P<0.05) in the extraction sockets treated with biphasic calcium phosphate at 8-week post-extraction than that in the other study groups. In this study, we demonstrated that the proposed animal model is useful to evaluate the bone formation after tooth extraction and the alveolar ridge conservation is feasible. The new bone formation and alveolar ridge preservation with bone graft after extraction of molar teeth, could result in the maintenance of sufficient bone volume to place an implant in an ideal restorative position without the need for ancillary implant site development procedures.


Compact Bragg grating with embedded metallic nano-structures.
A compact Bragg grating with embedded gapped metallic nano-structures is proposed and investigated theoretically. The Bragg grating consists of periodic planar metallic strips embedded in a dielectric waveguide. The grating exhibits distinct polarization characteristics due to its underlying working mechanisms of the metallic nano-strips. The grating can be considered as insulator-metal-insulator surface plasmonic polariton waveguide grating with improved light confinement for TM polarized waves. For the TE waves, significant field mismatch between metal and non-metal sections of the grating results in strong reflection. Comparison with the conventional deeply-etched grating on the same waveguide structures reveals interesting characteristics. It is concluded that the two types of grating structures share similar guidance, reflection and loss mechanisms for the TE modes. The spectral characteristics and their dependences on grating duty cycle are drastically different for the TM modes, mainly due to the SPP effect for the metal. Although the proposed grating performs slightly worse comparing to the deeply-etched grating for TE waves, its fabrication process should be easier since there will be no narrow trench (in sub-microns) deep-etching process (up to a few microns in depth) involved.


An information theoretic approach to insulin sensing by human kidney podocytes.
Podocytes are key components of the glomerular filtration barrier (GFB). They are insulin-responsive but can become insulin-resistant, causing features of the leading global cause of kidney failure, diabetic nephropathy. Insulin acts via insulin receptors to control activities fundamental to GFB integrity, but the amount of information transferred is unknown. Here we measure this in human podocytes, using information theory-derived statistics that take into account cell-cell variability. High content imaging was used to measure insulin effects on Akt, FOXO and ERK. Mutual Information (MI) and Channel Capacity (CC) were calculated as measures of information transfer. We find that insulin acts via noisy communication channels with more information flow to Akt than to ERK. Information flow estimates were increased by consideration of joint sensing (ERK and Akt) and response trajectory (live cell imaging of FOXO1-clover translocation). Nevertheless, MI values were always <1Bit as most information was lost through signaling. Constitutive PI3K activity is a predominant feature of the system that restricts the proportion of CC engaged by insulin. Negative feedback from Akt supressed this activity and thereby improved insulin sensing, whereas sensing was robust to manipulation of feedforward signaling by inhibiting PI3K, PTEN or PTP1B. The decisions made by individual podocytes dictate GFB integrity, so we suggest that understanding the information on which the decisions are based will improve understanding of diabetic kidney disease and its treatment.


Dosimetric evaluation and comparison of different RF exposure apparatuses used in human volunteer studies.
The aim of this study was to provide the information necessary to enable the comparison of exposure conditions in different human volunteer studies published by the research groups at the Universities of Turku, Swinburne, and Zurich. The latter applied a setup optimized for human volunteer studies in the context of risk assessment while the first two applied a modified commercial mobile phone for which detailed dosimetric data were lacking. While the Zurich Setup exposed the entire cortex of the target hemisphere, the other two setups resulted in only very localized exposure of the upper cheek, and concentrated on a limited area of the middle temporal gyrus just above the ear. The resulting peak spatial SAR averaged over 1 g of the cortex was 0.19 W/kg of the Swinburne Setup, and 0.31 W/kg for the Turku Setup, compared to 1 W/kg for the Zurich Setup. The average exposure of the thalamus was 5% and 9% of the Zurich Setup results for the Swinburne and Turku Setups, respectively. In general, the phone-based setup results in only reasonably defined exposures in a very limited area around the maximum exposure; the exposure of the rest of the cortex was low, and may vary greatly as a function of the setup, position, and local anatomy. The analysis confirms the need for a carefully designed exposure setup that exposes the relevant brain areas to a well-defined level in human volunteer studies, and shows that studies can only be properly compared and replicated if sufficiently detailed dosimetric information is available.


Mutagenesis and analysis of genetic mutations in the GC-rich KISS1 receptor sequence identified in humans with reproductive disorders.
The kisspeptin receptor (KISS1R) is a G protein-coupled receptor recognized as the trigger of puberty and a regulator of reproductive competence in adulthood (1,2,3). Inactivating mutations in KISS1R identified in patients have been associated with iodiopathic hypogonadotropic hypogonadism (IHH) (1,2) and precocious puberty (4). Functional studies of these mutants are crucial for our understanding of the mechanisms underlying the regulation of reproduction by this receptor as well as those shaping the disease outcomes, which result from abnormal KISS1R signaling and function. However, the highly GC-rich sequence of the KISS1R gene makes it rather difficult to introduce mutations or amplify the gene encoding this receptor by PCR. Here we describe a method to introduce mutations of interest into this highly GC-rich sequence that has been used successfully to generate over a dozen KISS1R mutants in our laboratory. We have optimized the PCR conditions to facilitate the amplification of a range of KISS1R mutants that include substitutions, deletions or insertions in the KISS1R sequence. The addition of a PCR enhancer solution, as well as of a small percentage of DMSO were especially helpful to improve amplification. This optimized procedure may be useful for other GC-rich templates as well. The expression vector encoding the KISS1R is been used to characterize signaling and function of this receptor in order to understand how mutations may change KISS1R function and lead to the associated reproductive phenotypes. Accordingly, potential applications of KISS1R mutants generated by site-directed mutagenesis can be illustrated by many studies (1,4,5,6,7,8). As an example, the gain-of-function mutation in the KISS1R (Arg386Pro), which is associated with precocious puberty, has been shown to prolong responsiveness of the receptor to ligand stimulation (4) as well as to alter the rate of degradation of KISS1R (9). Interestingly, our studies indicate that KISS1R is degraded by the proteasome, as opposed to the classic lysosomal degradation described for most G protein-coupled receptors (9). In the example presented here, degradation of the KISS1R is investigated in Human Embryonic Kidney Cells (HEK-293) transiently expressing Myc-tagged KISS1R (MycKISS1R) and treated with proteasome or lysosome inhibitors. Cell lysates are immunoprecipitated using an agarose-conjugated anti-myc antibody followed by western blot analysis. Detection and quantification of MycKISS1R on blots is performed using the LI-COR Odyssey Infrared System. This approach may be useful in the study of the degradation of other proteins of interest as well.


Stretching of BDT-gold molecular junctions: thiol or thiolate termination?
It is often assumed that the hydrogen atoms in the thiol groups of a benzene-1,4-dithiol dissociate when Au-benzene-1,4-dithiol-Au junctions are formed. We demonstrate, by stability and transport property calculations, that this assumption cannot be made. We show that the dissociative adsorption of methanethiol and benzene-1,4-dithiol molecules on a flat Au(111) surface is energetically unfavorable and that the activation barrier for this reaction is as high as 1 eV. For the molecule in the junction, our results show, for all electrode geometries studied, that the thiol junctions are energetically more stable than their thiolate counterparts. Due to the fact that density functional theory (DFT) within the local density approximation (LDA) underestimates the energy difference between the lowest unoccupied molecular orbital and the highest occupied molecular orbital by several electron-volts, and that it does not capture the renormalization of the energy levels due to the image charge effect, the conductance of the Au-benzene-1,4-dithiol-Au junctions is overestimated. After taking into account corrections due to image charge effects by means of constrained-DFT calculations and electrostatic classical models, we apply a scissor operator to correct the DFT energy level positions, and calculate the transport properties of the thiol and thiolate molecular junctions as a function of the electrode separation. For the thiol junctions, we show that the conductance decreases as the electrode separation increases, whereas the opposite trend is found for the thiolate junctions. Both behaviors have been observed in experiments, therefore pointing to the possible coexistence of both thiol and thiolate junctions. Moreover, the corrected conductance values, for both thiol and thiolate, are up to two orders of magnitude smaller than those calculated with DFT-LDA. This brings the theoretical results in quantitatively good agreement with experimental data.


Maternal depression and primary healthcare use for children: a population-based cohort study in Denmark.
Depression is a common mental illness worldwide. The offspring of a mother with depression has higher risk of developing mental and physical illness. This study aimed to investigate the association between the timing of maternal depression and the use of primary health care for the offspring. A population-based birth cohort study in Danish primary care using Danish national registers. All Danish children born between 1 January 2000 and 31 December 2013 (n = 869 140 children) were included in the study. The primary outcome was number and type of annual contacts with the GP. The secondary outcome was specific services used by the GP to assess inflammatory and infectious disease in the children. Exposure was maternal depression of four categories: non-depressed, recent, previous, and past depression. The association was expressed as adjusted incidence rate ratios (IRR) with 95% confidence intervals (CI). Maternal depression was associated with a higher use of primary health care for all three categories of depression. The strongest association was found for children of a mother with recent depression; they had 16% more contacts than children of a non-depressed mother (adjusted IRR = 1.16, 95% CI = 1.15 to 1.17), and 19-24% more positive infectious-related tests were found in this group. Exposure to maternal depression was associated with a significantly higher use of primary health care for the offspring for all exposure categories. These findings reveal that healthcare use is higher for the offspring exposed to maternal depression, even several years after expected remission. The higher ratio of positive tests indicates that exposed children are ill with infectious disease more often.


Revolution of Alzheimer Precision Neurology. Passageway of Systems Biology and Neurophysiology.
The Precision Neurology development process implements systems theory with system biology and neurophysiology in a parallel, bidirectional research path: a combined hypothesis-driven investigation of systems dysfunction within distinct molecular, cellular, and large-scale neural network systems in both animal models as well as through tests for the usefulness of these candidate dynamic systems biomarkers in different diseases and subgroups at different stages of pathophysiological progression. This translational research path is paralleled by an "omics"-based, hypothesis-free, exploratory research pathway, which will collect multimodal data from progressing asymptomatic, preclinical, and clinical neurodegenerative disease (ND) populations, within the wide continuous biological and clinical spectrum of ND, applying high-throughput and high-content technologies combined with powerful computational and statistical modeling tools, aimed at identifying novel dysfunctional systems and predictive marker signatures associated with ND. The goals are to identify common biological denominators or differentiating classifiers across the continuum of ND during detectable stages of pathophysiological progression, characterize systems-based intermediate endophenotypes, validate multi-modal novel diagnostic systems biomarkers, and advance clinical intervention trial designs by utilizing systems-based intermediate endophenotypes and candidate surrogate markers. Achieving these goals is key to the ultimate development of early and effective individualized treatment of ND, such as Alzheimer's disease. The Alzheimer Precision Medicine Initiative (APMI) and cohort program (APMI-CP), as well as the Paris based core of the Sorbonne University Clinical Research Group "Alzheimer Precision Medicine" (GRC-APM) were recently launched to facilitate the passageway from conventional clinical diagnostic and drug development toward breakthrough innovation based on the investigation of the comprehensive biological nature of aging individuals. The APMI movement is gaining momentum to systematically apply both systems neurophysiology and systems biology in exploratory translational neuroscience research on ND.


Cerec goes inLab--the metamorphosis of the system.
In spite of its underlying conception as being a chairside procedure, the Cerec procedure is now establishing itself in the dental laboratory as well. Based on hard core infiltration ceramic technology, it allows the production of crown caps and bridge frames held with a single attachment. The scan machine of the existing Cerec product family forms the technical basis of the new Cerec inLab instrument. Numerous modifications have been carried out. In addition, not only the magnification of the implementable work piece size, but also the changes in the scan process have been modified, which allows a higher resolution and the introduction of a new, very delicate milling bur. The software has been designed to be very user-friendly; the design of caps and frames has been greatly simplified. Without basic changes to the clinical process used for metal-ceramic, the smooth transition to full-ceramic restorations is possible for the dentist. The distinct separation of Cerec units for the dentist and Cerec inLab as a laboratory machine makes sense simply because of the high proportion of dental-technical steps. The new instrument can be seen as a further step toward the integration of CAD/CAM technology in the field of dentistry, of equal benefit to dentists, dental technicians, and patients.


Integrated Photonic Nanofences: Combining Subwavelength Waveguides with an Enhanced Evanescent Field for Sensing Applications.
Photonic nanofences consisting of high aspect ratio polymeric optical subwavelength waveguides have been developed for their application into photonic sensing devices. They are up to millimeter long arrays of 250 nm wide and 6 μm high ridges produced by an advanced lithography process on a silicon substrate enabling their straightforward integration into complex photonic circuits. Both simulations and experimental results show that the overlap of the evanescent fields propagating from each photonic nanofence allows for the formation of an effective waveguide that confines the overall evanescent field within its limits. This permits a high interaction with the surrounding medium which can be larger than 90% of the total guided light intensity (approximately 20000 times larger than the evanescent field of a standard waveguide with equivalent dimensions). In this work, we not only investigate the photonic properties of these structures but also demonstrate their successful integration into a photonic sensor. An absorbance-based sensor for the determination of lead in water samples is therefore achieved by the combination of the photonic nanofences with an ion-sensitive optical membrane. The experimental results for lead detection in water show a sensitivity of 0.102 AU/decade, and a linear range between 10(-6) M and 10(-2) M Pb(II). A detection limit as low as 7.3 nM has been calculated according to IUPAC for a signal-to-noise ratio of 3.


Extreme Tetralogy of Fallot With Polycythemia in a Ferret (Mustela putorius furo).
A 5-month-old, intact male ferret (Mustela putorius furo) was presented with apathy, tachypnea and exercise intolerance. On initial physical examination, tachypnea and cyanosis were the two primary clinical signs detected. The complete blood count also revealed a severe polycythemia, with lymphopenia, neutrophilia and eosinophilia. Further diagnostic imaging tests were performed. The thoracic radiographs revealed cardiomegaly and a diffuse alveolar pulmonary pattern. Consequently, an echocardiography was performed and showed an interventricular septal defect, pulmonic artery occlusion, overriding of the aorta and right ventricle hypertrophy. All abnormalities were compatible with a tetralogy of Fallot (TOF). As a treatment plan, a phlebotomy was performed and the ferret was posteriorly maintained with oxygen and fluid therapy. The following day, the hematocrit decreased 11% and the respiratory distress improved. Four days later, the ferret got clinically worse and was euthanized. A necropsy was performed and confirmed the presence of an extreme TOF with a diffused interstitial pneumonia secondary to Pneumocystis carinii. To the author's knowledge, this is the first report of an extreme tetralogy of Fallot with polycythemia in a ferret.


The symptom structure of panic attacks in depressed and anxious patients.
This study examined the panic symptom profiles of three diagnostic groups: those with panic disorder and no history of major depression; those with panic disorder with a history of major depressive episode but no current depression; and those current major depression with panic disorder. Patients were compared on the frequency of specific panic attack symptoms based on structured interview responses. The symptom profiles of all three groups were significantly correlated. The patients with past and current depressive episodes had the most similar symptom structure.


Alpha 2-adrenergic and opiate receptor blockade. Synergistic effects on anxiety in healthy subjects.
To evaluate interactions between the opiate and adrenergic systems in healthy humans, concomitant administration of the opiate antagonist, naloxone hydrochloride, and the alpha 2-adrenergic receptor antagonist, yohimbine hydrochloride, was compared with the administration of placebo and of each drug separately. A synergistic effect of the combination (larger than the sum of the effects of the two drugs separately) was observed on subject ratings of nervousness, anxiety, tremors, palpitations, nausea, hot and cold flashes, and increased plasma cortisol concentrations. In addition, following the combination, each of the male subjects studied reported a full penile erection lasting at least 60 minutes, an effect not reported when each drug was given separately. These results demonstrate that interactions between the opiate the adrenergic systems have important implications for our understanding of the cause and treatment of anxiety disorders and male impotence.


Focal adhesion kinase (pp125FAK) cleavage and regulation by calpain.
Focal adhesion kinase (125 kDa form; pp125FAK) is a widely expressed non-receptor tyrosine kinase that is implicated in integrin-mediated signal transduction. We have identified a novel means of pp 125FAK regulation in human platelets, in which this kinase undergoes sequential proteolytic modification from the native 125 kDa form to 90, 45 and 40 kDa fragments in thrombin-, collagen- and ionophore A23187-stimulated platelets. The proteolysis of pp125FAK was prevented by pretreating platelets with the calpain inhibitors calpeptin or calpain inhibitor-1, and was reproduced in vitro by incubating immunoprecipitated pp125FAK with purified calpain. Proteolysis of pp125FAK resulted in a dramatic reduction in its autokinase activity and led to its dissociation from the cytoskeletal fraction of platelets. These studies define a novel signal-terminating role for calpain, wherein proteolytic modification of pp125FAK attenuates its autokinase activity and induces its subcellular relocation within the cell.


Improving Physician Recommendations for Human Papillomavirus Vaccination: The Role of Professional Organizations.
To address low human papillomavirus (HPV) vaccination coverage, the American Academy of Family Physicians (AAFP) and the American Academy of Pediatrics (AAP) have launched national campaigns encouraging physicians to deliver strong HPV vaccine recommendations. We surveyed family physicians and pediatricians to examine the impact of these efforts on physicians' recommendation practices. A national sample of family physicians and pediatricians (n = 776) completed our online survey in 2014. The survey assessed reach, content, and influence of AAFP and AAP communications about HPV vaccination. The survey also assessed quality of physicians' communication practices for recommending HPV vaccination. Forty-seven percent of family physicians reported receiving information on HPV vaccination from AAFP, whereas 62% of pediatricians reported receiving information from AAP. Among physicians reached by AAFP or AAP, most reported receiving the message to give strong recommendations to adolescent boys (71%) and girls (78%). Although receiving information was not associated with HPV vaccine recommendation quality, receiving the message to give strong recommendations correlated with delivering higher-quality recommendations for boys (odds ratio, 4.19, 95% confidence interval, 2.64-6.64) and girls (odds ratio, 3.15, 95% confidence interval, 1.91-5.18). Over half of physicians reported improving their HPV vaccine communication after receiving information from AAFP (69%) or AAP (53%). Our findings suggest that it is important for AAFP and AAP to communicate the need for strong HPV vaccine recommendations. Given that many physicians reported improving their recommendation practices, professional organizations stand to contribute to increasing HPV vaccination coverage, but they will likely need to increase the intensity of quality improvement efforts to do so.


Imaging patients with acute abdominal pain.
Acute abdominal pain may be caused by a myriad of diagnoses, including acute appendicitis, diverticulitis, and cholecystitis. Imaging plays an important role in the treatment management of patients because clinical evaluation results can be inaccurate. Performing computed tomography (CT) is most important because it facilitates an accurate and reproducible diagnosis in urgent conditions. Also, CT findings have been demonstrated to have a marked effect on the management of acute abdominal pain. The cost-effectiveness of CT in the setting of acute appendicitis was studied, and CT proved to be cost-effective. CT can therefore be considered the primary technique for the diagnosis of acute abdominal pain, except in patients clinically suspected of having acute cholecystitis. In these patients, ultrasonography (US) is the primary imaging technique of choice. When costs and ionizing radiation exposure are primary concerns, a possible strategy is to perform US as the initial technique in all patients with acute abdominal pain, with CT performed in all cases of nondiagnostic US. The use of conventional radiography has been surpassed; this examination has only a possible role in the setting of bowel obstruction. However, CT is more accurate and more informative in this setting as well. In cases of bowel perforation, CT is the most sensitive technique for depicting free intraperitoneal air and is valuable for determining the cause of the perforation. Imaging is less useful in cases of bowel ischemia, although some CT signs are highly specific. Magnetic resonance (MR) imaging is a promising alternative to CT in the evaluation of acute abdominal pain and does not involve the use of ionizing radiation exposure. However, data on the use of MR imaging for this indication are still sparse. http://radiology.rsna.org/content/253/1/31/suppl/DC1.


A Review of Antimicrobial Therapy for Infectious Uveitis of the Posterior Segment.
Treatment of infectious posterior uveitis represents a therapeutic challenge for ophthalmologists. The eye is a privileged site, maintained by blood ocular barriers, which limits penetration of systemic antimicrobials into the posterior segment. In addition, topical and subconjunctival therapies are incapable of producing sufficient drug concentrations, intraocularly. Posterior infectious uveitis can be caused by bacteria, virus, fungi, or protozoa. Mode of treatment varies greatly based on the infectious etiology. Certain drugs have advantages over others in the treatment of infectious uveitis. Topical and systemic therapies are often employed in the treatment of ocular infection, yet the route of treatment can have limitations based on penetration, concentration, and duration. The introduction of intravitreal antimicrobial therapy has advanced the management of intraocular infections. Being able to bypass blood-ocular barriers allows high drug concentrations to be delivered directly to the posterior segment with minimal systemic absorption. However, because the difference between the therapeutic and the toxic doses of some antimicrobial drugs falls within a narrow concentration range, intravitreal therapy could be associated with ocular toxicity risks. In many cases of infectious uveitis, combination of intravitreal and systemic therapies are necessary. In this comprehensive review, the authors aimed at reviewing clinically relevant data regarding intraocular and systemic antimicrobial therapy for posterior segment infectious uveitis. The review also discussed the evolving trends in intraocular treatment, and elaborated on antibiotic pharmacokinetics and pharmacodynamics, efficacy, and adverse effects.


Secondary lymphedema in the mouse tail: Lymphatic hyperplasia, VEGF-C upregulation, and the protective role of MMP-9.
Disturbances in the microcirculation can lead to secondary lymphedema, a common pathological condition that, despite its frequency, still lacks a cure. Lymphedema is clinically well described, but while the genetic underpinnings that cause lymphatic malformations and primary lymphedema are being discovered, the pathophysiology and pathobiology of secondary lymphedema remain poorly understood, partly due to the lack of well-described experimental models. Here, we provide a detailed characterization of secondary lymphedema in the mouse tail and correlate the evolution of tissue swelling to changes in tissue architecture, infiltration of immune cells, deposition of lipids, and proliferation and morphology of the lymphatic vessels. We show that sustained swelling leads to lymphatic hyperplasia and upregulation of vascular endothelial growth factor (VEGF)-C, which may exacerbate the edema because the hyperplastic vessels are poorly functional. The onset of lymphatic hyperplasia occurred prior to the onset of lipid accumulation and peak VEGF-C expression. Langerhans dendritic cells were seen in the dermis migrating from the epidermis to the lymphatic capillaries in edematous tissue. Furthermore, these results were consistent between two different normal mouse strains, but swelling was significantly greater in a matrix metalloproteinase (MMP)-9 null strain. Thus, by characterizing this highly reproducible model of secondary lymphedema, we conclude that VEGF-C upregulation and lymphatic hyperplasia resulting from dermal lymphatic ligation and lymphedema leads to decreased drainage function and that MMP-9 may be important in counteracting tissue swelling.


Oral health status and oral hygiene habits of an adult Turkish population on dialysis.
The present study was undertaken to evaluate the oral health status and oral hygiene habits of dialysis patients. A total of 145 patients on dialysis were evaluated in two centres. Evaluation included oral examination of soft and hard tissues and completion of a questionnaire that was used to gather information about demographic, socioeconomic, medical and dental status and oral hygiene attitudes of patients. Awareness of the importance of oral health was also determined by four yes/no questions within this questionnaire. Of the 145 patients, 89 (61.4%) were presently candidates for renal transplantation. Nineteen patients (13.1%) were edentulous and nearly half of the patients (n=69, 47.6%) had 15 or fewer remaining teeth. Dentate patients who regularly brushed twice a day and flossed were few (n=18, 14.3%; n=3, 2.4%, respectively). In addition, plaque score was more than 50% in most of the dentate patients (93.7%). Gingival bleeding was reported by about two thirds of dentate patients (67.5%). None of the patients were referred to a dentist or were attending regular dental care. The percentage of patients who were aware of oral cavity-related infections and importance of oral hygiene following renal transplantation was very low (20.7% and 9.7%, respectively). Poor oral health, unsatisfactory daily oral hygiene habits and insufficient awareness of the importance of oral health is frequent among Turkish dialysis patients. Dentists and nephrologists should work in close cooperation in order to raise awareness and encourage regular dental controls.


Robotic bilateral inguinal lymphadenectomy in penile cancer, development of a technique without robot repositioning: a case report.
Inguinal lymphadenectomy is the treatment of choice for patients with penile cancer and inguinal lymph node metastases. We describe the performance of the robotic bilateral inguinal lymphadenectomy technique without repositioning the robot in a patient with penile carcinoma and high risk for nodal metastases and no palpable lymph nodes. A 64-year-old male patient was diagnosed with penile cancer (TNM: T3 N 0 M 0) and underwent a total penectomy with perineal urethrostomy. We performed a robotic bilateral inguinal lymphadenectomy four weeks after the penectomy. The entire procedure was performed with the robot-assisted technique. The operative time, median estimated blood loss, and hospital stay was 360 min, 100 ml (50 ml in the right side and 150 ml in the left side), and three days, respectively. Metastatic nodes were present in both inguinal regions, with a yield of 19 lymph nodes on the right and 14 on the left. The patient presented with a left-side lymphocele that was drained at follow-up. No other complications were reported. Robotic bilateral inguinal lymphadenectomy secondary to penile cancer is feasible, safe, and provides a good performance. Prospective studies are required to include a larger number of patients and long-term monitoring to assess the results of this procedure in comparison with open and laparoscopic techniques.


A bioresponsive hydrogel tuned to chondrogenesis of human mesenchymal stem cells.
Cartilage tissue engineering aims to replace damaged or diseased tissue with a functional regenerate that restores joint function. Scaffolds are used to deliver cells and facilitate tissue development, but they can also interfere with the structural assembly of the cartilage matrix. Biodegradable scaffolds have been proposed as a means to improve matrix deposition and the biomechanical properties of neocartilage. The challenge is designing scaffolds with appropriate degradation rates, ideally such that scaffold degradation is proportional to matrix deposition. In this study, we developed a bioresponsive hydrogel with cell-mediated degradation aligned to the chondrogenic differentiation of human mesenchymal stem cells (hMSCs). We identified matrix metalloproteinase 7 (MMP7) as an enzyme with a temporal expression pattern that corresponded with cartilage development. By embedding MMP7 peptide substrates within a poly(ethylene glycol) diacrylate backbone, we built MMP7-sensitive hydrogels with distinct degradation rates. When MMP7-sensitive scaffolds were compared with nondegradable scaffolds in vitro, photoencapsulated hMSCs produced neocartilage constructs with more extensive collagenous matrices, as demonstrated through immunohistochemistry and biochemical quantification of matrix molecules. Furthermore, these changes translated into an increased dynamic compressive modulus. This work presents a practical strategy for designing biomaterials uniquely tuned to individual biological processes.


The effects of improving the mesothelioma surveillance network on sensitivity, timeliness in reporting and asbestos exposure assessment.
In Italy, Mesothelioma Registries (MRs) have been established by law for the epidemiological surveillance of occupational cancers. MRs collect information about asbestos exposure of incident cases, through interviews. In the Emilia-Romagna region, MR was implemented in 1996 and extended its network of health professionals who report suspected mesothelioma in 2001 and 2007. This study evaluated the impact of the extension of the network on MR sensitivity and timeliness. Mesothelioma cases were analysed in three subsequent periods: 1996-2001 (before any network extension), 2002-2007 (after first extension) and 2008-2014 (after second extension). Sensitivity was evaluated by the proportion of cases directly reported by the network out of the total number of incident cases; reporting and interview timeliness were assessed by median times between diagnosis and, respectively, reporting and interview. Pleural mesothelioma reporting timeliness was also evaluated by use of quantile regression models, stratified by diagnostic certainty and adjusted by sex and age. Sensitivity increased from 79.4% (1996-2001), to 89.0% (2002-2007) and to 91.4% (2008-2013). For mesothelioma with diagnostic certainty, we recorded considerably reduced reporting times from the 50th percentile on, whereas for uncertain mesothelioma relevant reductions were observed also in the lower percentiles. A reduced time to interview was observed too, which was more significant for uncertain cases. The proportion of patients directly interviewed increased from 33.5% (1996-2001), to 39.1% (2002-2007), to 49.5% (2008-2014). The extended network improved the MR sensitivity and allowed shorter reporting and interview times and more frequent patient interviews, thus improving accuracy of exposure definition.


The increasing dominance of teams in production of knowledge.
We have used 19.9 million papers over 5 decades and 2.1 million patents to demonstrate that teams increasingly dominate solo authors in the production of knowledge. Research is increasingly done in teams across nearly all fields. Teams typically produce more frequently cited research than individuals do, and this advantage has been increasing over time. Teams now also produce the exceptionally high-impact research, even where that distinction was once the domain of solo authors. These results are detailed for sciences and engineering, social sciences, arts and humanities, and patents, suggesting that the process of knowledge creation has fundamentally changed.


Relationships between physical qualities and key performance indicators during match-play in senior international rugby union players.
The use of physical tests to profile physical capabilities, and provide training direction to athletes is common practice. Likewise, in professional team sports, notational analysis codes the key contributions of each player during competition. Limited studies have however investigated relationships between physical capabilities and key performance indicators (KPIs) of rugby union match-play. Elite professional players, categorised as forwards (n = 15) or backs (n = 14), from an international rugby union squad (n = 29) undertook assessments of isometric mid-thigh pull (IMTP), bilateral and unilateral countermovement jumps (CMJ) and drop jumps (DJ; from 40 and 20 cm, respectively), and assessment of acceleration (10 m), a 5 m weighted sled drive, and a Yo-Yo intermittent recovery test level 1 (Yo-Yo IRTL1). Game statistics of the same players from 92 matches (~23 matches per player) during the 2014-15 season were analysed for effort and performance-based metrics. For forwards, Yo-Yo IRTL1 correlated significantly with; number of tackles made (r = 0.717), first three players at a ruck in both attack (r = 0.568) and defence (r = 0.581), number of effective rucks (r = 0.630), total possessions (r = 0.522), passes made (r = 0.651), percentage of carries over the gainline (r = 0.610), effective ruck success (r = 0.600), tackle success (r = 0.540), and the number of turnovers made (r = 0.518). Drop jump performance in forwards was associated with; the number of clean breaks (r = 0.558), dominant collisions (r = 0.589), and offloads (r = 0.594). For backs, the sled-drive test correlated with; number of carries (r = -0.751), first three players at an attacking ruck (r = -0.613), effective attacking rucks (r = -0.584), number of dominant collisions (r = -0.792) and offloads (r = -0.814). Likewise, for backs, IMTP peak force was related to; the number of possessions (r = 0.793), passes made (r = 0.792), effective attacking ruck percentage (r = 0.628), and the number of offloads (r = 0.621) whilst relative peak force correlated with; the percentage of carries over the gainline (r = 0.533), percent tackle success (r = 0.603) and effective attacking ruck percentage (r = 0.584). Regression analyses highlighted that only a small number of variables (i.e., carries, tackles, attacking and defensive first three at ruck) returned practically achievable changes (<20%) in physical qualities. In spite of this, and while leaving scope identification of further physical and/or performance predictors, greater strength, power and intermittent running performance were positively related to match-derived KPIs during competition. This may provide a basis for better integrating the strategies used by physical and technical performance-focused coaching staff to improve key performance indicators, and thus match performance, of rugby union players.


Identification of process-localized mRNAs from cultured rodent hippocampal neurons.
The regulated translation of localized mRNAs in neurons provides a mechanism for spatially restricting gene expression in a synapse-specific manner. To identify the population of mRNAs present in distal neuronal processes of rodent hippocampal neurons, we grew neurons on polycarbonate filters etched with 3 microm pores. Although the neuronal cell bodies remained on the top surface of the filters, dendrites, axons, and glial processes penetrated through the pores to grow along the bottom surface of the membrane where they could be mechanically separated from cell bodies. Quantitative PCR and immunochemical analyses of the process preparation revealed that it was remarkably free of somatic contamination. Microarray analysis of RNA isolated from the processes identified over 100 potentially localized mRNAs. In situ hybridization studies of 19 of these transcripts confirmed that all 19 were present in dendrites, validating the utility of this approach for identifying dendritically localized transcripts. Many of the identified mRNAs encoded components of the translational machinery and several were associated with the RNA-binding protein Staufen. These findings indicate that there is a rich repertoire of mRNAs whose translation can be locally regulated and support the emerging idea that local protein synthesis serves to boost the translational capacity of synapses.


Alpha Particle Detection Using Alpha-Induced Air Radioluminescence: A Review and Future Prospects for Preliminary Radiological Characterisation for Nuclear Facilities Decommissioning.
The United Kingdom (UK) has a significant legacy of nuclear installations to be decommissioned over the next 100 years and a thorough characterisation is required prior to the development of a detailed decommissioning plan. Alpha radiation detection is notoriously time consuming and difficult to carry out due to the short range of alpha particles in air. Long-range detection of alpha particles is therefore highly desirable and this has been attempted through the detection of secondary effects from alpha radiation, most notably the air-radioluminescence caused by ionisation. This paper evaluates alpha induced air radioluminescence detectors developed to date and looks at their potential to develop a stand-off, alpha radiation detector which can be used in the nuclear decommissioning field in daylight conditions to detect alpha contaminated materials.


Streptococcus ovis sp. nov., isolated from sheep.
Seven strains of an unknown Gram-positive catalase-negative chain-forming coccus-shaped organism isolated from clinical specimens from sheep were characterized by phenotypic and molecular taxonomic methods. Comparative 16S rRNA gene sequencing studies demonstrated that the bacterium represents a new sub-line within the genus Streptococcus. The unknown bacterium was readily distinguished from recognized streptococcal species by biochemical tests and electrophoretic analysis of whole-cell proteins. Based on phylogenetic and phenotypic evidence, it is proposed that the unknown bacterium be classified as Streptococcus ovis sp. nov. The type strain of Streptococcus ovis is CCUG 39485T (= LMG 19174T).


Effect of fatigue on maximal inspiratory pressure-flow capacity.
The inspiratory muscles can be fatigued by repetitive contractions characterized by high force (inspiratory resistive loads) or high velocities of shortening (hyperpnea). The effects of fatigue induced by inspiratory resistive loaded breathing (pressure tasks) or by eucapnic hyperpnea (flow tasks) on maximal inspiratory pressure-flow capacity and rib cage and diaphragm strength were examined in five healthy adult subjects. Tasks consisted of sustaining an assigned breathing frequency, duty cycle, and either a "pressure-time product" of esophageal pressure (for the pressure tasks) or peak inspiratory flow rate (for the flow tasks). Esophageal pressure was measured during maximal inspiratory efforts against a closed glottis (Pesmax), maximal transdiaphragmatic pressure was measured during open-glottis expulsive maneuvers (Pdimax), and maximal inspiratory flow (VImax) was measured during maximal inspiratory efforts with no added external resistance before and after fatiguing pressure and flow tasks. The reduction in Pesmax) with pressure fatigue (-25 +/- 7%) was significantly greater than the change in Pesmax with flow fatigue (-8 +/- 8%, P less than 0.01). In contrast, the reductions in Pdimax (-11 +/- 8%) and VImax (-16 +/- 3%) with flow fatigue were greater than the changes in Pdimax (-0.6 +/- 4%, P less than 0.05) or VImax (-3 +/- 4%, P less than 0.05) with pressure fatigue. We conclude that respiratory muscle performance is dependent not only on the presence of fatigue but whether fatigue was induced by pressure tasks or flow tasks. The specific impairment of Pesmax and not of Pdimax or flow with pressure fatigue may reflect selective fatigue of the rib cage muscles.(ABSTRACT TRUNCATED AT 250 WORDS)


Why are the 2-oxoacid dehydrogenase complexes so large? Generation of an active trimeric complex.
The four-component polypeptides of the 2-oxoacid dehydrogenase complex from the thermophilic archaeon Thermoplasma acidophilum assemble to give an active multienzyme complex possessing activity with the branched-chain 2-oxoacids derived from leucine, isoleucine and valine, and with pyruvate. The dihydrolipoyl acyl-transferase (E2) core of the complex is composed of identical trimer-forming units that assemble into a novel 42-mer structure comprising octahedral and icosahedral geometric aspects. From our previously determined structure of this catalytic core, the inter-trimer interactions involve a tyrosine residue near the C-terminus secured in a hydrophobic pocket of an adjacent trimer like a ball-and-socket joint. In the present study, we have deleted the five C-terminal amino acids of the E2 polypeptide (IIYEI) and shown by equilibrium centrifugation that it now only assembles into a trimeric enzyme. This was confirmed by SAXS analysis, although this technique showed the presence of approximately 20% hexamers. The crystal structure of the trimeric truncated E2 core has been determined and shown to be virtually identical with the ones observed in the 42-mer, demonstrating that removal of the C-terminal anchor does not significantly affect the individual monomer or trimer structures. The truncated E2 is still able to bind both 2-oxoacid decarboxylase (E1) and dihydrolipoamide dehydrogenase (E3) components to give an active complex with catalytic activity similar to the native multienzyme complex. This is the first report of an active mini-complex for this enzyme, and raises the question of why all 2-oxoacid dehydrogenase complexes assemble into such large structures.


Skeleton of the Oldest Known Pinniped, Enaliarctos mealsi.
A nearly complete skeleton of the archaic pinniped Enaliarctos, found in late Oligocene or early Miocene rocks (approximately 23 million years old) of California, provides new evidence on the origin of pinnipeds. Enaliarctos retains many primitive features expected in the hypothesized common ancestor of pinnipeds. Skeletal modifications seen in Enaliarctos document swimming adaptations and indicate that pinnipeds primitively used the axial skeleton and both fore and hindflippers as sources of propulsion. Elongate hindlimbs with prominent bony processes (reflecting powerful musculature) suggest that Enaliarctos was more active on land than modern pinnipeds.


The diagnostic utility of desmin. A study of 584 cases and review of the literature.
The diagnostic utility of several antibodies against desmin and their optimal staining conditions have not been systematically evaluated. Sections of paraffin-embedded tissues from 584 cases were stained with a monoclonal antibody against desmin (Clone DER 11 from DAKO), using an avidin-biotin-peroxidase technique. The results were tabulated and compared with those from previous reports. The following observations were made: (1) When pronase digestion was performed before staining, desmin was equally demonstrable in tissues fixed in formalin, Zenker's, Bouin's, or B5 fixative; however, desmin staining was lost or significantly diminished in tissues fixed in absolute ethyl alcohol. In contrast, when pronase was not used, a positive staining was demonstrated only in tissue fixed in absolute ethyl alcohol. (2) Positive staining was found in normal muscle (92 of 92 cases), leiomyoma (12 of 13), rhabdomyoma (6 of 6), rhabdomyosarcoma (31 of 31), leiomyosarcoma (18 of 26). (3) Desmin was never found in epithelia, normal mesenchymal tissue other than muscle, tumors stimulating rhabdomyosarcoma, and epithelial tumors. (4) A positive staining was documented in 1 tumor (a fibrous histiocytoma) of 42 benign predominantly spindle cell tumors and in 8 of 89 predominantly spindle cell sarcomas. (5) Desmin was never documented in myoepithelial cells but stained myofibroblasts in 2 of 12 examples of granulation tissue and in 29 of 67 samples containing tumor-associated desmoplasia. The authors' data on the diagnostic sensitivity and specificity of the evaluated antibody should improve the use of desmin in diagnostic pathology.