Soft tissue sleeve approach to open reduction and internal fixation of proximal phalangeal fractures.
Proximal phalangeal fractures often lead to motion loss at the proximal interphalangeal joint primarily because of adhesions between the zone 4 extensor tendon and the fracture site. This most commonly manifests as an extensor lag at the proximal interphalangeal joint, but can also include incomplete recovery of flexion range. The occurrence of the fracture alone likely generates some degree of insurmountable motion loss. Additional motion loss may be iatrogenic based on the specific surgical technique and rehabilitation strategy. It has been promoted that percutaneous pinning methods, as opposed to open fixation methods, produce less of a motion deficit. Although percutaneous pinning solves many fractures of the proximal phalanx, some fracture patterns will not be adequately treated by percutaneous pinning and will benefit from direct open reduction and internal fixation. Spiral fractures with a significant rotational deformity are well treated by lag screws. Comminuted fractures are well treated by plate fixation. When performing such fixations, the soft tissue approach to the fracture site plays a substantial role in determining the ultimate functional recovery. This article covers the specific technical details of optimizing soft tissue management when performing open fixation for proximal phalanx fractures.


Preparing plants for improved cold tolerance by priming.
Cold is a major stressor, which limits plant growth and development in many parts of the world, especially in the temperate climate zones. A large number of experimental studies has demonstrated that not only acclimation and entrainment but also the experience of single short stress events of various abiotic or biotic kinds (priming stress) can improve the tolerance of plants to chilling temperatures. This process, called priming, depends on a stress "memory". It does not change cold sensitivity per se but beneficially modifies the response to cold and can last for days, months, or even longer. Elicitor factors and antagonists accumulate due to increased biosynthesis or decreased degradation either during or after the priming stimulus. Comparison of priming studies investigating improved tolerance to chilling temperatures highlighted key regulatory functions of ROS/RNS and antioxidant enzymes, plant hormones, especially jasmonates, salicylates, and abscisic acid, and signalling metabolites, such as β- and γ-aminobutyric acid (BABA and GABA) and melatonin. We conclude that these elicitors and antagonists modify local and systemic cold tolerance by integration into cold-induced signalling cascades.


Repair rate and durability of video assisted minimally invasive mitral valve surgery.
Minimally invasive mitral valve surgery (MIMVS) through a right lateral thoracotomy has emerged as another option in cardiac surgery. We present our single center experience with regards to repair rate and durability of the repair in patients undergoing MIMVS. Between January 2009 and December 2012 a total of 312 consecutive patients underwent MIMVS. Baseline characteristics, operative variables, postoperative outcomes, and follow-up information about survival, major adverse cardiac and cerebrovascular event (MACCE)-free survival, valve competence, and freedom from reoperation were collected. Mean age was 61 ± 13.1 years, median Euroscore 4 (0-13). The predominant valve pathology was degenerative disease (96.2%) with regurgitation (95.2%) and atrial fibrillation (23.1%). Bypass time was 147 ± 46, cross-clamp time 94 ± 32 minutes. All degenerative valves planned for repair (n = 281) were successfully repaired except for two (0.7%) resulting in a 99.3% repair rate. Eight patients (2.8%) had to be converted to sternotomy. Median ventilation time was 6.6 hours (0 to 460), ICU stay one day (0 to 96), and hospital stay eight days (3 to 61). Most of the patients did not need red blood cell transfusion (78.2%). One patient died in hospital due to acute respiratory distress syndrome (0.3%) and five suffered a permanent stroke (1.6%). In 294 patients (94.5%) follow-up was completed for a median of 22 months (0 to 53) with 95.5 ± 1.2% overall survival, 96.8 ± 1.2%, MACCE-free survival, and 94.8 ± 0.1% freedom from reoperation. MIMVS is a safe approach with low morbidity and mortality that allows a high and durable repair rate with low reoperation rates and favorable short-term event-free survival.


Frequency dependence of respiratory resistance in healthy children.
We measured in 130 (61 girls) children aged 3--14 yr respiratory resistance (Rrs), with the oscillation technique, between 4 and 9 Hz. Rrs, at both 4 and 9 Hz, decreased as a function of height (r = 0.74, P less than 0.001). No statistical difference was found between boys and girls. Frequency dependence of resistance (Rrs 4 Hz-Rrs 9 Hz) was found in children at all ages, and decreased with increasing height (r = 0.50, P less than 0.001). We suggest that frequency dependence of resistance in children can be explained on the basis of an increased peripheral resistance, which produces an asynchronous distribution of tidal volume between dead space and lung parenchyma. During growth peripheral resistance decreases and Rrs bcome less frequency dependent to reach at about 15--16 yr independency of frequence.


Dihomo-γ-linolenic acid inhibits several key cellular processes associated with atherosclerosis.
Atherosclerosis and its complications are responsible for one in three global deaths. Nutraceuticals show promise in the prevention and treatment of atherosclerosis but require an indepth understanding of the mechanisms underlying their actions. A previous study showed that the omega-6 fatty acid, dihomo-γ-linolenic acid (DGLA), attenuated atherosclerosis in the apolipoprotein E deficient mouse model system. However, the mechanisms underlying such protective effects of DGLA are poorly understood and were therefore investigated. We show that DGLA attenuates chemokine-driven monocytic migration together with foam cell formation and the expression of key pro-atherogenic genes induced by three pro-inflammatory cytokines in human macrophages. The effect of DGLA on interferon-γ signaling was mediated via inhibition of signal transducer and activator of transcription-1 phosphorylation on serine 727. In relation to anti-foam cell action, DGLA inhibits modified LDL uptake by both macropinocytosis and receptor-mediated endocytosis, the latter by reduction in expression of two key scavenger receptors (SR-A and CD36), and stimulates cholesterol efflux from foam cells. DGLA also improves macrophage mitochondrial bioenergetic profile by decreasing proton leak. Gamma-linolenic acid and prostaglandin E1, upstream precursor and key metabolite respectively of DGLA, also acted in an anti-atherogenic manner. The actions of DGLA extended to other key atherosclerosis-associated cell types with attenuation of endothelial cell proliferation and migration of smooth muscle cells in response to platelet-derived growth factor. This study provides novel insights into the molecular mechanisms underlying the anti-atherogenic actions of DGLA and supports further assessments on its protective effects on plaque regression in vivo and in human trials.


Involvement of Three Esterase Genes from Panonychus citri (McGregor) in Fenpropathrin Resistance.
The citrus red mite, Panonychus citri (McGregor), is a major citrus pest with a worldwide distribution and an extensive record of pesticide resistance. However, the underlying molecular mechanism associated with fenpropathrin resistance in this species have not yet been reported. In this study, synergist triphenyl phosphate (TPP) dramatically increased the toxicity of fenpropathrin, suggesting involvement of carboxylesterases (CarEs) in the metabolic detoxification of this insecticide. The subsequent spatiotemporal expression pattern analysis of PcE1, PcE7 and PcE9 showed that three CarEs genes were all over-expressed after insecticide exposure and higher transcripts levels were observed in different field resistant strains of P. citri. Heterologous expression combined with 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetra-zolium bromide (MTT) cytotoxicity assay in Spodoptera frugiperda (Sf9) cells revealed that PcE1-, PcE7- or PcE9-expressing cells showed significantly higher cytoprotective capability than parental Sf9 cells against fenpropathrin, demonstrating that PcEs probably detoxify fenpropathrin. Moreover, gene silencing through the method of leaf-mediated dsRNA feeding followed by insecticide bioassay increased the mortalities of fenpropathrin-treated mites by 31% (PcE1), 27% (PcE7) and 22% (PcE9), respectively, after individual PcE gene dsRNA treatment. In conclusion, this study provides evidence that PcE1, PcE7 and PcE9 are functional genes mediated in fenpropathrin resistance in P. citri and enrich molecular understanding of CarEs during the resistance development of the mite.


Going from "paper and pen" to ICT systems: Perspectives on managing the change process.
Lack of participation from staff when developing information and communication technologies (ICT) has been shown to lead to negative consequences and might be one explanation for failure. Management during development processes has rarely been empirically studied, especially when introducing ICT systems in a municipality context. To describe and interpret experiences of the management during change processes where ICT was introduced among staff and managers in elderly care. A qualitative interpretive method was chosen for this study and content analysis for analyzing the interviews. "Clear focus-unclear process" demonstrated that focus on ICT solutions was clear but the process of introducing the ICT was not. "First-line managers receiving a system of support" gave a picture of the first-line manager as not playing an active part in the projects. First-line managers and staff described "Low power to influence" when realizing that for some reasons, they had not contributed in the change projects. "Low confirmation" represented the previous and present feelings of staff not being listened to. Lastly, "Reciprocal understanding" pictures how first-line managers and staff, although having some expectations on each other, understood each other's positions. Empowerment could be useful in creating an organization where critical awareness and reflection over daily practice becomes a routine.


Once-daily OROS hydromorphone ER compared with placebo in opioid-tolerant patients with chronic low back pain.
This multicenter, double-blind, placebo-controlled study using a randomized withdrawal design evaluated the efficacy and safety of once-daily OROS hydromorphone ER in the treatment of opioid-tolerant patients with chronic moderate-to-severe low back pain (LBP). The primary efficacy assessment was mean change in pain intensity based on patient diary Numeric Rating Scale (NRS) scores from baseline to final visit of the 12-week double-blind phase. Secondary endpoints included mean change from baseline to each visit in patient diary NRS scores; and office NRS scores; time to treatment failure; Patient Global Assessment; rescue medication use; and Roland Morris Disability Questionnaire total scores. ClinicalTrials.gov NCT00549042. For the primary outcome measure, hydromorphone ER significantly reduced pain intensity compared to placebo (p < 0.001). Median diary NRS score change from baseline to endpoint was significantly lower for OROS [corrected] hydromorphone ER (0.2 units) compared to placebo (1.6 units). [corrected] A significantly higher proportion of hydromorphone ER (60.6%) vs. placebo (42.9%) patients had at least a 30% reduction in diary NRS pain score from screening to endpoint (p < 0.01). Hydromorphone ER was well-tolerated, although 60 (13%) discontinued during the enrichment phase for adverse events and more active (9, 6.7%) than placebo (4, 3.0%) patients discontinued treatment for adverse events during the randomized phase. These results provide evidence for the efficacy and safety of hydromorphone ER in opioid-tolerant patients with chronic moderate-to-severe LBP. Potential limitations include the shortened dose-conversion/titration phase, limiting the daily allowable dose of hydromorphone ER to 64 mg, and the allowance of limited rescue medication throughout the entire double-blind phase. Other trial design elements such as the use of an enrichment phase and the inclusion of only opioid tolerant patients may limit the generalizability of these results.


Association between circulating vitamin K1 and coronary calcium progression in community-dwelling adults: the Multi-Ethnic Study of Atherosclerosis.
Animal studies have shown that vitamin K treatment reduced vascular calcification, but human data are limited. We determined the association between vitamin K status and coronary artery calcium (CAC) progression in the Multi-Ethnic Study of Atherosclerosis by using a case-cohort design. Serum phylloquinone (vitamin K1) was measured in 296 participants with extreme CAC progression and 561 randomly selected participants without extreme CAC progression; all subjects had baseline and follow-up CAC measures (mean follow-up: 2.5 y). A serum vitamin K1 concentration was considered low at <1.0 nmol/L (the distribution median). Outcomes were replicated by using post hoc per-protocol analyses of a vitamin K1 supplementation trial. The OR (95% CI) for extreme CAC progression for subjects with low serum vitamin K1 compared with subjects without extreme CAC progression was 1.34 (0.94, 1.90; NS) when adjusted for demographics and confounders. A significant interaction between low vitamin K1 and antihypertension medication use was detected (P = 0.016). Hypertension medication users with low serum vitamin K1 were more likely to have extreme CAC progression than were medication users without extreme CAC progression [OR (95% CI): 2.37 (1.38, 4.09)]. In replication, baseline antihypertensive medication users in the supplementation group had less CAC progression than did those in the control group [adjusted mean ± SEM of the 3-y CAC change was +5 ± 20 Agatston units (AU) in the vitamin K1 group (n = 40) and +44 ± 13 AU in the placebo group (n = 49); P < 0.01]. Although the point estimate of our primary analysis suggests low serum vitamin K1 is associated with greater CAC progression, the difference was NS. Low serum vitamin K1 was significantly associated with CAC progression in antihypertension medication users, which, to our knowledge, is a novel finding conditionally replicated by using an independent sample. Intervention trials are needed to determine whether improving serum vitamin K1 reduces CAC progression, especially in hypertensive individuals. This trial was registered at clinicaltrials.gov as NCT00183001.


Wasting, ischemia, and lymphoid abnormalities in mice expressing T cell-targeted human tumor necrosis factor transgenes.
To evaluate the biologic potential of T cell-specific TNF production in vivo, we have generated transgenic mice constitutively expressing TNF in their T cell compartment. This was achieved by placing a wild-type or a 3'-UTR modified fragment of the human TNF gene under the influence of the T cell-specific, locus control region of the human CD2 gene. Transgenic mice that express human TNF mRNA in T cells develop marked histologic and cellular changes locally in their lymphoid organs and a lethal wasting syndrome associated with widespread vascular thrombosis and tissue necrosis. The extent of pathologic changes and their time of onset appear to reflect levels of transgene expression. Thus, transgenic lines that express the transgene at high levels show both lymphoid organ and systemic abnormalities with wasting. In one transgenic line, mice express lower levels of the transgene and develop normally despite pronounced local lymphoid organ defects, confirming in vivo, the differential potential of localized and systemic TNF action. All pathologic changes could be neutralized by the administration of mAb specific for human TNF. These results demonstrate the important role of T cell-specific TNF production in the development of specific pathology and provide a means by which to evaluate the role of TNF in thymocyte development. Transgenic mice that express TNF constitutively in the T cell compartment offer a unique in vivo system by which to analyze the molecular character of systemic vs contact-dependent and paracrine modes of TNF action. Furthermore, given the species-specific nature of the mouse p75 TNF receptor, it is assumed that the pathology induced by human TNF in these transgenic mice is associated exclusively with p55 TNF receptor signaling. Conceivably, the differential contribution of each of the two TNF receptors in thymus development and TNF-mediated disease can be assessed by comparison of the biologic potential of human vs mouse TNF in the transgenic system developed.


On the consistency of personality types across adulthood: latent profile analyses in two large-scale panel studies.
Consistency and change in personality were analyzed by examining personality types across adulthood and old age using data from 2 nationally representative panel studies from Germany (N = 14,718; 16-82 years) and Australia (N = 8,315; 15-79 years). In both samples, the Big Five personality traits were measured twice across a period of 4 years. Latent profile analyses and latent profile transition analyses revealed 4 main findings: First, solutions with 3 (in the German sample) or 4 (in the Australian sample) personality types were found to be most interpretable. Second, measurement invariance tests revealed that these personality types were consistent across all age groups but differed slightly between men and women. Third, age was related to the number of individuals classified within each personality type. Namely, there were more resilients and fewer undercontrollers in older compared with younger age groups. Fourth, there was strong consistency of personality type membership across a period of 4 years in both genders and most age cohorts. Comparatively less consistency across time was found for undercontrollers and individuals in old age. Taken together, these findings show that in the 2 nations studied here, personality types were highly consistent across gender, age, and time.


The Src homology 2 domain-containing adapter protein B (SHB) regulates mouse oocyte maturation.
SHB (Src homology 2 domain-containing adapter protein B) is involved in receptor tyrosine kinase signaling. Mice deficient in the Shb gene have been found to exhibit a transmission ratio distortion with respect to inheritance of the Shb null allele among offspring and this phenomenon was linked to female gamete production. Consequently, we postulated that Shb plays a role for oocyte biology and thus decided to investigate oocyte formation, meiotic maturation, and early embryo development in relation to absence of the Shb gene. Oogenesis was apparently accelerated judging from the stages of oocyte development on fetal day 18.5 and one week postnatally in Shb -/- mice; but in adulthood ovarian follicle maturation was impaired in these mice. Completion of meiosis I (first polar body extrusion) was less synchronized, with a fraction of oocytes showing premature polar body extrusion in the absence of Shb. In vitro fertilization of mature oocytes isolated from Shb +/+, +/- and -/- mice revealed impaired early embryo development in the -/- embryos. Moreover, the absence of Shb enhanced ERK (extracellular-signal regulated kinase) and RSK (ribosomal S6 kinase) signaling in oocytes and these effects were paralleled by an increased ribosomal protein S6 phosphorylation and activation. It is concluded that SHB regulates normal oocyte and follicle development and that perturbation of SHB signaling causes defective meiosis I and early embryo development.


Abnormal anther development and high sporopollenin synthesis in benzotriazole treated male sterile Helianthus annuus L.
Foliar application of 1.5% benzotriazole induced 100% pollen sterility in H. annuus. Pollen abortion in treated plants was mainly associated with abnormal behaviour of tapetum. A limited number of anther locule showed early degeneration of tapetum followed by disintegration of sporogenous tissues. On the other hand, some locules showed normal development of tapetum at initial stages. However, this tapetum exhibited degenerated and non-functional cell organelles. In both these situations tapetum failed to provide proper nourishment to developing microspores. The ultrastructure of both tapetum and microspores is different from that of control material with irregularities of exine deposition, endopolyploidy of tapetal nuclei and an alteration of organelle composition being correlated with sterility. Pollen grains thus developed were devoid of nucleus and cell organelles and were complete sterile.


The INHANCE consortium: toward a better understanding of the causes and mechanisms of head and neck cancer.
The International Head and Neck Cancer Epidemiology (INHANCE) consortium is a collaboration of research groups leading large epidemiology studies to improve the understanding of the causes and mechanisms of head and neck cancer. The consortium includes investigators of 35 studies who have pooled their data on 25 500 patients with head and neck cancer (i.e., cancers of the oral cavity, oropharynx, hypopharynx, and larynx) and 37 100 controls. The INHANCE analyses have confirmed that tobacco use and alcohol intake are key risk factors of these diseases and have provided precise estimates of risk and dose response, the benefit of quitting, and the hazard of smoking even a few cigarettes per day. Other risk factors include short height, lean body mass, low education and income, and a family history of head and neck cancer. Risk factors are generally similar for oral cavity, pharynx, and larynx, although the magnitude of risk may vary. Some major strengths of pooling data across studies include more precise estimates of risk and the ability to control for potentially confounding factors and to examine factors that may interact with each other. The INHANCE consortium provides evidence of the scientific productivity and discoveries that can be obtained from data pooling projects.


Effects of homeopathic arsenicum album, nosode, and gibberellic acid preparations on the growth rate of arsenic-impaired duckweed (Lemna gibba L.).
This study evaluated the effects of homeopathically potentized Arsenicum album, nosode, and gibberellic acid in a bioassay with arsenic-stressed duckweed (Lemna gibba L.). The test substances were applied in nine potency levels (17x, 18x, 21x-24x, 28x, 30x, 33x) and compared with controls (unsuccussed and succussed water) regarding their influence on the plant's growth rate. Duckweed was stressed with arsenic(V) for 48 h. Afterwards, plants grew in either potentized substances or water controls for 6 days. Growth rates of frond (leaf) area and frond number were determined with a computerized image analysis system for different time intervals (days 0-2, 2-6, 0-6). Five independent experiments were evaluated for each test substance. Additionally, five water control experiments were analyzed to investigate the stability of the experimental setup (systematic negative control experiments). All experiments were randomized and blinded. The test system exhibited a low coefficient of variation (approximately equal to 1%). Unsuccussed and succussed water did not result in any significant differences in duckweed growth rate. Data from the control and treatment groups were pooled to increase statistical power. Growth rates for days 0-2 were not influenced by any homeopathic preparation. Growth rates for days 2-6 increased after application of potentized Arsenicum album regarding both frond area (p < 0.001) and frond number (p < 0.001), and by application of potentized nosode (frond area growth rate only, p < 0.01). Potencies of gibberellic acid did not influence duckweed growth rate. The systematic negative control experiments did not yield any significant effects. Thus, false-positive results can be excluded with high certainty. To conclude, the test system with L. gibba impaired by arsenic(V) was stable and reliable. It yielded evidence for specific effects of homeopathic Arsenicum album preparations and it will provide a valuable tool for future experiments that aim at revealing the mode of action of homeopathic preparations. It may also be useful to investigate the influence of external factors (e.g., heat, electromagnetic radiation) on the effects of homeopathic preparations.


Synchronous core-needle biopsy and microwave ablation for highly suspicious malignant pulmonary nodule via a coaxial cannula.
This study aimed to evaluate the safety and feasibility of computed tomography (CT)-guided synchronous percutaneous core-needle biopsy and microwave ablation (MWA) for highly suspicious malignant pulmonary nodules. This retrospective study evaluated medical records of 54 consecutive patients (mean age, 65.5 ± 11.2 years) with 62 highly suspicious malignant pulmonary nodules who synchronously underwent percutaneous core-needle biopsy and MWA via a coaxial cannula (Group A) or sequentially underwent these procedures (Group B) from September 2016 to November 2017. All patients were followed up for at least 6 months after MWA. The safety and feasibility of synchronous core-needle biopsy and MWA were analyzed by comparing clinical data, technical success rate, complication, and curative effect per nodule with those of sequential procedures. Technical success rates were 100% in both groups. The pneumothorax rate was 29.6% (8/27) in Group A and 57.1% (20/35) in Group B, which was statistically different (P = 0.031). In Group A, hemoptysis and pleural effusion rates were 22.2% (6/27), and in Group B, the corresponding rates were 28.6% (10/35) and 20.0% (7/35), respectively. No postprocedural pulmonary artery pseudoaneurysm, bronchopleural fistula, or needle-tract tumor seeding developed in both groups. After 6 months' follow-up, the effective rates (complete + partial response) in both groups were 100%. Synchronous core-needle biopsy and MWA via a coaxial cannula is technically safe and feasible in the management of highly suspicious malignant pulmonary nodules, and this procedure has lesser complications and similar effects (both 100% effective treatment) compared with sequential procedures.


Genepool variation in genus Glycine subgenus Soja revealed by polymorphic nuclear and chloroplast microsatellites.
A combination of nuclear and chloroplast simple sequence repeats (SSRs) have been used to investigate the levels and pattern of variability detected in Glycine max and G. soja genotypes. Based on the analysis of 700 soybean genotypes with 115 restriction fragment length polymorphism (RFLP) probes, 12 accessions were identified that represent 92% of the allelic variability detected in this genepool. These 12 core genotypes together with a sample of G. max and G. soja accessions were evaluated with 11 nuclear SSRs that detected 129 alleles. Compared with the other G. max and G. soja genotypes sampled, the core genotypes represent 40% of the allelic variability detected with SSRs. Despite the multi-allelic nature of soybean SSRs, dendrograms representing phenetic relationships between accessions clustered according to their subspecies origin. In addition to biparentally inherited nuclear SSRs, two uniparentally (maternally) transmitted chloroplast SSRs were also studied. A total of seven haplotypes were identified, and diversity indices of 0.405 +/- 0.088 and 0.159 +/- 0.071 were obtained for the two chloroplast SSRs. The availability of polymorphic SSR loci in the chloroplast genome provides new opportunities to investigate cytonuclear interactions in plants.


Functional thalamocortical synapse reorganization from subplate to layer IV during postnatal development in the reeler-like mutant rat (shaking rat Kawasaki).
Transient synapse formation between thalamic axons and subplate neurons is thought to be important in thalamocortical targeting. Shaking rat Kawasaki (SRK), having reversed cortical layering similarly observed in reeler mouse, provides an interesting model system to test this idea. The spatial and temporal pattern of excitation was investigated using optical recording with voltage-sensitive dyes in thalamocortical slice preparations from SRK. At postnatal day 0 (P0), a strong optical response was elicited within the superplate of the SRK in the cell layer corresponding to subplate in wild-type (WT) rats. By P3, this response rapidly descended into deep cortical layers comprised of layer IV cells, as identified with 5-bromo-2'-deoxyuridine birthdating at embryonic day 17. During the first 3 postnatal days, both the subplate and cortical plate responses were present, but by P7, the subplate response was abolished. Tracing individual axons in SRK revealed that at P0-P3, a large number of thalamocortical axons reach the superplate, and by P7-P10, the ascending axons develop side branches into the lower or middle cortical layers. Synaptic currents were also demonstrated in WT subplate cells and in SRK superficial cortical cells using whole-cell recording. These currents were elicited monosynaptically, because partial AMPA current blockade did not modify the latencies. These results suggest that the general developmental pattern of synapse formation between thalamic axons and subplate (superplate) neurons in WT and SRK is very similar, and individual thalamic arbors in cortex are considerably remodeled during early postnatal development to find layer IV equivalent neurons.


Advantages of using tetrahydrofuran-water as mobile phases in the quantitation of cyclosporin A in monkey and rat plasma by liquid chromatography-tandem mass spectrometry.
A new analytical method is described here for the quantitation of anti-inflammatory drug cyclosporin A (CyA) in monkey and rat plasma. The method used tetrahydrofuran (THF)-water mobile phases to elute the analyte and internal standard, cyclosporin C (CyC). The gradient mobile phase program successfully eluted CyA into a sharp peak and therefore improved resolution between the analyte and possible interfering materials compared with previously reported analytical approaches, where CyA was eluted as a broad peak due to the rapid conversion between different conformers. The sharp peak resulted from this method facilitated the quantitative calculation as multiple smoothing and large number of bunching factors were not necessary. The chromatography in the new method was performed at 30 degrees C instead of 65-70 degrees C as reported previously. Other advantages of the method included simple and fast sample extraction-protein precipitation, direct injection of the extraction supernatant to column for analysis, and elimination of evaporation and reconstitution steps, which were needed in solid phase extraction or liquid-liquid extraction reported before. This method is amenable to high-throughput analysis with a total chromatographic run time of 3 min. This approach has been verified as sensitive, linear (0.977-4000 ng/mL), accurate and precise for the quantitation of CyA in monkey and rat plasma. However, compared with the usage of conventional mobile phases, the only drawback of this approach was the reduced detection response from the mass spectrometer that was possibly caused by poor desolvation in the ionization source. This is the first report to demonstrate the advantages of using THF-water mobile phases to elute CyA in liquid chromatography.


Curcumin and tetrahydrocurcumin both prevent osteoarthritis symptoms and decrease the expressions of pro-inflammatory cytokines in estrogen-deficient rats.
Menopausal symptoms are associated with inflammation. Curcumin is a well-known anti-inflammatory bioactive compound from turmeric whereas tetrahydrocurcumin (THC) is a major metabolite of curcumin that may have different efficacies. However, they have not been studied for anti-menopausal symptoms and anti-osteoarthritis effects. We compared the efficacies of curcumin and THC for preventing postmenopausal and osteoarthritis symptoms in ovariectomized (OVX) obese rats with monoiodoacetate (MIA) injections into the right knee to generate a similar pathology as osteoarthritis. OVX rats were provided a 45 % fat diet containing either (1) 0.4 % curcumin (curcumin), (2) 0.4 % THC, (3) 30 μg/kg body weight 17β-estradiol + 0.4 % dextrin (positive control), (4) 0.4 % dextrin (placebo; control), or (5) 0.4 % dextrin with no MIA injection (normal control) for 4 weeks. At the beginning of the fifth week, OVX rats were given articular injections of MIA or normal-control saline into the right knee and the assigned diets were provided for an additional 3 weeks. Curcumin and THC had similar efficacies for skin tail temperature in OVX rats whereas THC, but not curcumin, prevented glucose intolerance, which might be involved in exacerbating osteoarthritis. Both protected against osteoarthritis symptoms and pain-related behaviors better than 17β-estradiol treatment in estrogen-deficient rats. Curcumin and THC prevented the deterioration of articular cartilage compared to control. They also maintained lean body mass and lowered fat mass as much as 17β-estradiol treatment. The improvement in osteoarthritis symptoms was associated with decreased gene expressions of matrix metalloproteinase (MMP)3 and MMP13 and tumor necrosis factor-α, interleukin (IL)1β, and IL6 in the articular cartilage. THC and curcumin are effective for treating postmenopausal and osteoarthritis symptoms in OVX rats with MIA-induced osteoarthritis-like symptoms and may have potential as interventions for menopausal and osteoarthritic symptoms in humans.


B Cell IgD Deletion Prevents Alveolar Bone Loss Following Murine Oral Infection.
Periodontal disease is one of the most common infectious diseases of humans. Immune responses to infection trigger loss of alveolar bone from the jaw and eventual tooth loss. We investigated the contribution of B cell IgD to alveolar bone loss by comparing the response of B cell normal BALB/cJ mice and IgD deficient BALB/c-Igh-5(-/-J) mice to oral infection with Porphyromonas gingivalis, a gram-negative periodontopathic bacterium from humans. P. gingivalis-infected normal mice lost bone. Specific antibody to P. gingivalis was lower and oral colonization was higher in IgD deficient mice; yet bone loss was completely absent. Infection increased the proportion of CD69(+) activated B cells and CD4(+) T cells in immune normal mice compared to IgD deficient mice. These data suggest that IgD is an important mediator of alveolar bone resorption, possibly through antigen-specific coactivation of B cells and CD4(+) T cells.


The Proteome of Prostate Cancer Bone Metastasis Reveals Heterogeneity with Prognostic Implications.
Purpose: Bone is the most predominant site of distant metastasis in prostate cancer, and patients have limited therapeutic options at this stage.Experimental Design: We performed a system-wide quantitative proteomic analysis of bone metastatic prostate tumors from 22 patients operated to relieve spinal cord compression. At the time of surgery, most patients had relapsed after androgen-deprivation therapy, while 5 were previously untreated. An extended cohort of prostate cancer bone metastases (n = 65) was used for immunohistochemical validation.Results: On average, 5,067 proteins were identified and quantified per tumor. Compared with primary tumors (n = 26), bone metastases were more heterogeneous and showed increased levels of proteins involved in cell-cycle progression, DNA damage response, RNA processing, and fatty acid β-oxidation; and reduced levels of proteins were related to cell adhesion and carbohydrate metabolism. Within bone metastases, we identified two phenotypic subgroups: BM1, expressing higher levels of AR canonical targets, and mitochondrial and Golgi apparatus resident proteins; and BM2, with increased expression of proliferation and DNA repair-related proteins. The two subgroups, validated by the inverse correlation between MCM3 and prostate specific antigen immunoreactivity, were related to disease prognosis, suggesting that this molecular heterogeneity should be considered when developing personalized therapies.Conclusions: This work is the first system-wide quantitative characterization of the proteome of prostate cancer bone metastases and a valuable resource for understanding the etiology of prostate cancer progression. Clin Cancer Res; 24(21); 5433-44. ©2018 AACR.


Second-phase hepatitis C virus RNA decline during telaprevir-based therapy increases with drug effectiveness: implications for treatment duration.
Hepatitis C virus (HCV) RNA decay during antiviral therapy is characterized by a rapid first phase, followed by a slower second phase. The current understanding of viral kinetics attributes the magnitude of the first phase of decay to treatment effectiveness, whereas the second phase of decay is attributed to the progressive loss of infected cells. Here, we analyzed data from 44 patients treated with telaprevir, a potent HCV protease inhibitor. Using a viral kinetic model that accounts for the pharmacokinetics of telaprevir, we found the second-phase slope of viral decline to be strongly correlated with treatment effectiveness and to be roughly four-fold more rapid than has been reported with interferon-based therapies. Because telaprevir is not known to increase the death rate of infected cells, our results suggest that the second-phase slope of viral decline is driven not only by the death of infected cells, but may also involve other mechanisms, such as a treatment-effectiveness-dependent degradation of intracellular viral RNA. As a result of the enhanced viral decay caused by the high antiviral effectiveness of telaprevir, we predict that if drug resistance could be avoided by using an appropriate combination of antiviral agents, treatment duration needed to clear HCV might be dramatically shortened. Indeed, we predict that in 95% of fully compliant patients, the last virus particle should be eliminated by week 7 of therapy. If the remaining infected hepatocytes act as a potential reservoir for the renewal of infection, no more than 10 weeks of treatment should be sufficient to clear the infection in 95% of fully compliant patients. However, if patients miss doses, treatment duration would need to be extended.


Pulsed pumping of semiconductor disk lasers.
Efficient operation of semiconductor disk lasers is demonstrated using uncooled and inexpensive 905nm high-power pulsed semiconductor pump lasers. Laser emission, with a peak power of 1.7W, is obtained from a 2.3mum semiconductor disk laser. This is seven times the power achieved under continuous pumping. Analysis of the time-dependent spectral characteristics of the laser demonstrate that significant device heating occurs over the 100-200ns duration of the pumping pulse - finite element modelling of the thermal processes is undertaken in support of these data. Spectral narrowing to below 0.8nm is obtained by using an intra-cavity birefringent filter.


Biphasic targeting and cleavage furrow ingression directed by the tail of a myosin II.
Cytokinesis in animal and fungal cells utilizes a contractile actomyosin ring (AMR). However, how myosin II is targeted to the division site and promotes AMR assembly, and how the AMR coordinates with membrane trafficking during cytokinesis, remains poorly understood. Here we show that Myo1 is a two-headed myosin II in Saccharomyces cerevisiae, and that Myo1 localizes to the division site via two distinct targeting signals in its tail that act sequentially during the cell cycle. Before cytokinesis, Myo1 localization depends on the septin-binding protein Bni5. During cytokinesis, Myo1 localization depends on the IQGAP Iqg1. We also show that the Myo1 tail is sufficient for promoting the assembly of a "headless" AMR, which guides membrane deposition and extracellular matrix remodeling at the division site. Our study establishes a biphasic targeting mechanism for myosin II and highlights an underappreciated role of the AMR in cytokinesis beyond force generation.


The efficacy of psychodynamic psychotherapy.
Empirical evidence supports the efficacy of psychodynamic therapy. Effect sizes for psychodynamic therapy are as large as those reported for other therapies that have been actively promoted as "empirically supported" and "evidence based." In addition, patients who receive psychodynamic therapy maintain therapeutic gains and appear to continue to improve after treatment ends. Finally, nonpsychodynamic therapies may be effective in part because the more skilled practitioners utilize techniques that have long been central to psychodynamic theory and practice. The perception that psychodynamic approaches lack empirical support does not accord with available scientific evidence and may reflect selective dissemination of research findings.


On the dispersion management of fluorite whispering-gallery mode resonators for Kerr optical frequency comb generation in the telecom and mid-infrared range.
Optical whispering gallery mode (WGM) resonators have been very attracting platforms for versatile Kerr frequency comb generations. We report a systematic study on the material dispersion of various optical materials that are capable of supporting quality factors above 109. Using an analytical approximation of WGM resonant frequencies in disk resonators, we investigate the effect of the geometry and transverse mode order on the total group-velocity dispersion (GVD). We demonstrate that the major radii and the radial mode indices play an important role in tailoring the GVD of WGM resonators. In particular, our study shows that in WGM disk-resonators, the polar families of modes have very similar GVD, while the radial families of modes feature dispersion values that can differ by up to several orders of magnitude. The effect of these giant dispersion shifts are experimentally evidenced in Kerr comb generation with magnesium fluoride. From a more general perspective, this critical feature enables to push the zero-dispersion wavelength of fluorite crystals towards the mid-infrared (mid-IR) range, thereby allowing for efficient Kerr comb generation in that spectral range. We show that barium fluoride is the most interesting crystal in this regard, due to its zero dispersion wavelength (ZDW) at 1.93 μm and an optimal dispersion profile in the mid-IR regime. We expect our results to facilitate the design of different platforms for Kerr frequency comb generations in both telecommunication and mid-IR spectral ranges.


The prevalence of lip vermilion morphological traits in a 15-year-old population.
The aim of this study is to describe and categorize the various lip vermilion morphological traits, prevalence, and associations present in a 15-year-old population The instrument we developed was checked for the reliability of assessment. The study is a cross-sectional study of lip vermilion morphology in 2,246 fifteen-year-old children. Three-dimensional surface laser scans were taken of the children, and a classification scale was derived and assessed for intra- and inter-reliability. The prevalence of lip vermilion morphological traits and associations was identified. Certain features are found to be likely to be associated with each other: thin philtrum and smooth/deep indentation of the philtrum; wide philtrum and smooth/normal gradient, flat/absent Cupid's bow and smooth/normal/indentation near nose, V-shaped Cupid's bow and indentation in middle/deep groove; thin upper lip vermilion and deep groove; thick upper lip vermilion and smooth philtrum; acute nasolabial angle and upper lip double border and thick upper lip vermilion; obtuse nasolabial angle and thin upper lip vermilion. In conclusion, we present a new method to identify the various morphological features of the vermilion of the lips. The prevalence of morphological traits has been identified in this 15-year-old population. In addition, the association of lip traits has been reported. This classification scale may be utilized to ascertain prevalence of features amongst other ethnic groups, to gain knowledge of what is considered to be attractive features, and to determine genotype/phenotype associations.


FGF-8 stimulates neuronal differentiation through FGFR-4a and interferes with mesoderm induction in Xenopus embryos.
The role of fibroblast growth factors (FGFs) in neural induction is controversial [1,2]. Although FGF signalling has been implicated in early neural induction [3-5], a late role for FGFs in neural development is not well established. Indeed, it is thought that FGFs induce a precursor cell fate but are not able to induce neuronal differentiation or late neural markers [6-8]. It is also not known whether the same or distinct FGFs and FGF receptors (FGFRs) mediate the effects on mesoderm and neural development. We report that Xenopus embryos expressing ectopic FGF-8 develop an abundance of ectopic neurons that extend to the ventral, non-neural, ectoderm, but show no ectopic or enhanced notochord or somitic markers. FGF-8 inhibited the expression of an early mesoderm marker, Xbra, in contrast to eFGF, which induced ectopic Xbra robustly and neuronal differentiation weakly. The effect of FGF-8 on neurogenesis was blocked by dominant-negative FGFR-4a (DeltaXFGFR-4a). Endogenous neurogenesis was also blocked by DeltaXFGFR-4a and less efficiently by dominant-negative FGFR-1 (XFD), suggesting that it depends preferentially on signalling through FGFR-4a. The results suggest that FGF-8 and FGFR-4a signalling promotes neurogenesis and, unlike other FGFs, FGF-8 interferes with mesoderm induction. Thus, different FGFs show specificity for mesoderm induction versus neurogenesis and this may be mediated, at least in part, by the use of distinct receptors.


Attenuation of proteasome-induced proteolysis in skeletal muscle by {beta}-hydroxy-{beta}-methylbutyrate in cancer-induced muscle loss.
Loss of skeletal muscle is an important determinant of survival in patients with cancer-induced weight loss. The effect of the leucine metabolite beta-hydroxy-beta-methylbutyrate (HMB) on the reduction of body weight loss and protein degradation in the MAC16 model of cancer-induced weight loss has been compared with that of eicosapentaenoic acid (EPA), a recognized inhibitor of protein degradation. HMB was found to attenuate the development of weight loss at a dose greater than 0.125 g/kg accompanied by a small reduction in tumor growth rate. When EPA was used at a suboptimal dose level (0.6 g/kg) the combination with HMB seemed to enhance the anticachectic effect. Both treatments caused an increase in the wet weight of soleus muscle and a reduction in protein degradation, although there did not seem to be a synergistic effect of the combination. Proteasome activity, determined by the "chymotrypsin-like" enzyme activity, was attenuated by both HMB and EPA. Protein expression of the 20S alpha or beta subunits was reduced by at least 50%, as were the ATPase subunits MSS1 and p42 of the 19S proteasome regulatory subunit. This was accompanied by a reduction in the expression of E2(14k) ubiquitin-conjugating enzyme. The combination of EPA and HMB was at least as effective or more effective than either treatment alone. Attenuation of proteasome expression was reflected as a reduction in protein degradation in gastrocnemius muscle of cachectic mice treated with HMB. In addition, HMB produced a significant stimulation of protein synthesis in skeletal muscle. These results suggest that HMB preserves lean body mass and attenuates protein degradation through down-regulation of the increased expression of key regulatory components of the ubiquitin-proteasome proteolytic pathway, together with stimulation of protein synthesis.


Effects of different discoloration challenges and whitening treatments on dental hard tissues and composite resin restorations.
To compare the relative effects of cigarette smoke (CS), electronic cigarette (EC), red wine, coffee, and soy sauce on the color of enamel, dentin, and composite resin restorations, as well as the effects of whitening treatments. Seventy premolars with composite restorations were exposed to CS, EC aerosol (a novel EC device with MESH™ technology [P4M3 version 1.0, Philip Morris International]), red wine, coffee, and soy sauce for 56 min/day for 15 days. Two whitening sessions with 6% and 35% hydrogen peroxide (H2O2) were performed on the exposed samples. Teeth exposed to CS and EC aerosol were also brushed with whitening toothpaste for 3 weeks. Color match of resin restorations was assessed, and color changes were compared after exposure and after whitening treatments. Discolorations in enamel, dentin, and composite resin were observed in the order of red wine > CS > soy sauce > coffee > EC. Color mismatch between enamel and resin restorations occurred only in red wine and CS groups. Brushing with whitening toothpaste removed discoloration caused by EC aerosol; H2O2 treatments were necessary to eliminate discolorations caused by coffee and soy sauce. Discolorations of dentin and resin restorations could not be completely removed by whitening treatments, and color mismatch remained in teeth exposed to red wine and CS. Red wine and CS cause significant tooth discoloration and color mismatch in enamel and resin restorations that are not reversible by whitening treatments. Tooth discoloration associated with EC aerosol was minimal and could be removed by brushing with whitening toothpaste. Red wine drinkers and cigarette smokers have increased risks for tooth discoloration and color mismatch between enamel and composite resin restorations. Whitening treatments may not be effective in correcting the color mismatch. Tooth discoloration associated with EC aerosol is minimal.


The role of attributions in self-poisoning.
Sixty patients who had taken an overdose of drugs within the previous 24 hours completed the Attributional Style Questionnaire and the Profile of Mood States scale. Their scores were compared with two age-matched samples: 31 medical patients and 71 non-patient controls. Overdose patients had significantly higher stable and global attributions for negative events than controls, but were no different in attribution for positive events. They were also more disturbed in their mood, but their level of mood disturbance did not correlate with levels of attribution. Thirty-two of the overdose group were followed up one week later, by which time there had been a reliable downward shift in mood. However, there was no corresponding reliable shift in attributions, which remained significantly higher than normal. These results suggest that attributions may be important in determining how self-poisoners cope with life-events, but the relationship with mood is complex; attributions are not simply mood-dependent, nor is mood level or recovery simply attribution-dependent.


Rapid, scalable and highly automated HLA genotyping using next-generation sequencing: a transition from research to diagnostics.
Human leukocyte antigen matching at allelic resolution is proven clinically significant in hematopoietic stem cell transplantation, lowering the risk of graft-versus-host disease and mortality. However, due to the ever growing HLA allele database, tissue typing laboratories face substantial challenges. In light of the complexity and the high degree of allelic diversity, it has become increasingly difficult to define the classical transplantation antigens at high-resolution by using well-tried methods. Thus, next-generation sequencing is entering into diagnostic laboratories at the perfect time and serving as a promising tool to overcome intrinsic HLA typing problems. Therefore, we have developed and validated a scalable automated HLA class I and class II typing approach suitable for diagnostic use. A validation panel of 173 clinical and proficiency testing samples was analysed, demonstrating 100% concordance to the reference method. From a total of 1,273 loci we were able to generate 1,241 (97.3%) initial successful typings. The mean ambiguity reduction for the analysed loci was 93.5%. Allele assignment including intronic sequences showed an improved resolution (99.2%) of non-expressed HLA alleles. We provide a powerful HLA typing protocol offering a short turnaround time of only two days, a fully integrated workflow and most importantly a high degree of typing reliability. The presented automated assay is flexible and can be scaled by specific primer compilations and the use of different 454 sequencing systems. The workflow was successfully validated according to the policies of the European Federation for Immunogenetics. Next-generation sequencing seems to become one of the new methods in the field of Histocompatibility.


Transposon tagging of tobacco mosaic virus resistance gene N: its possible role in the TMV-N-mediated signal transduction pathway.
Plants can recognize and resist invading pathogens by signaling the induction of rapid defense responses. Often these responses are mediated by single dominant resistance genes (R genes). The products of R genes have been postulated to recognize the pathogen and trigger rapid host defense responses. Here we describe isolation of the classical resistance gene N of tobacco that mediates resistance to the well-characterized pathogen tobacco mosaic virus (TMV). The N gene was isolated by transposon tagging using the maize Activator (Ac) transposon. We confirmed isolation of the N gene by complementation of the TMV-sensitive phenotype with a genomic DNA fragment. Sequence analysis of the N gene shows that it encodes a protein with an amino-terminal domain similar to that of the cytoplasmic domains of the Drosophila Toll protein and the interleukin 1 receptor in mammals, a putative nucleotide-binding site and 14 imperfect leucine-rich repeats. The presence of these functional domains in the predicted N gene product is consistent with the hypothesis that the N resistance gene functions in a signal transduction pathway. Similarities of N to Toll and the interleukin 1 receptor suggest a similar signaling mechanism leading to rapid gene induction and TMV resistance.


Fresh and cultured mouse islets differ in their response to nutrient stimulation.
Observing different kinetics of nutrient-induced insulin secretion in fresh and cultured islets under the same condition we compared parameters of stimulus secretion coupling in freshly isolated and 22-h-cultured NMRI mouse islets. Stimulation of fresh islets with 30 mM glucose after perifusion without nutrient gave a continuously ascending secretion rate. In 22-h-cultured islets the same protocol produced a brisk first phase followed by a moderately elevated plateau, a pattern regarded to be typical for mouse islets. This was also the response of cultured islets to the nutrient secretagogue alpha-ketoisocaproic acid, whereas the secretion of fresh islets increased similarly fast but remained strongly elevated. The responses of fresh and cultured islets to purely depolarizing stimuli (tolbutamide or KCl), however, were closely similar. Signs of apoptosis and necrosis were rare in both preparations. In cultured islets, the glucose-induced rise of the cytosolic Ca2+ concentration started from a lower value and was larger as was the increase of the ATP/ADP ratio. The prestimulatory level of mitochondrial reducing equivalents, expressed as the NAD(P)H/FAD fluorescence ratio, was lower in cultured islets, but increased more strongly than in fresh islets. When culture conditions were modified by replacing RPMI with Krebs-Ringer medium and FCS with BSA, the amount of released insulin varied widely, but the kinetics always showed a predominant first phase. In conclusion, the secretion kinetics of fresh mouse islets is more responsive to variations of nutrient stimulation than cultured islets. The more uniform kinetics of the latter may be caused by a different use of endogenous metabolites.


Early cortical facilitation for emotionally arousing targets during the attentional blink.
The present study aimed to investigate the time course of electrocortical facilitation for affectively arousing written words during the so-called 'attentional blink' (AB) period in a rapid serial visual presentation (RSVP) task. The AB refers to a period of reduced awareness for second-target stimuli following a first target by an interval of about 200-500 ms. Pleasant, neutral, and unpleasant written verbs were used as second targets in an 8.6-Hz RSVP paradigm that contained affectively neutral words as distractors. Replicating and extending behavioral studies, we expected that emotional second targets would be associated with better identification accuracy and greater electrocortical activity, compared with neutral targets. The steady-state visual evoked potential was recorded using 129 scalp electrodes. The time-varying energy at the presentation frequency of 8.6 Hz was extracted as a continuous measure of electrocortical activity related to the RSVP stream. Behavioral data showed that at an inter-target interval of 232 ms, the report for emotionally arousing (pleasant and unpleasant) words was more accurate than for neutral control words. This result was mirrored by the electrocortical response at posterior sensors, which showed rapid amplitude enhancement (120-270 ms after T2 onset) for pleasant and unpleasant targets specifically. The present data suggest that identification facilitation for emotionally arousing target words in the AB is related to rapid enhancement of sensory processing. Affectively arousing information is preferentially selected at the level of early perceptual analysis, leading to facilitation at later stages of processing, including consolidation in working memory and visual awareness.


Nucleophilicity Parameters of Arylsulfonyl-Substituted Halomethyl Anions.
The rates of the reactions of the arylsulfonyl-substituted carbanions carrying α-chloro and α-bromo substituents (1a-e) with quinone methides 2a-g and benzylidenemalonates 2h and 2i in DMSO were determined photometrically at 20 °C. The reactions were performed under pseudo-first-order conditions, and the second-order rate constants were obtained as the slopes of the plots of the pseudo-first-order rate constants versus the concentrations of the reactants used in excess. The second-order rate constants correlate linearly with the parameters E of the reference electrophiles according to the linear free energy relationship log k2(20 °C) = sN(N + E), which allowed us to derive the nucleophile-specific parameters N and sN of carbanions 1a-e. The resulting nucleophilicity parameters N (23 < N < 29) reveal the title compounds to be among the most reactive nucleophiles so far integrated on our comprehensive nucleophilicity scale.


Patterning the differentiation of C2C12 skeletal myoblasts.
Mammalian cells are sensitive to the physical properties of their micro-environment such as the stiffness and geometry of the substrate. It is known that the stiffness of the substrate plays a key role in the process of mammalian myogenesis. However, the effect of geometrical constraints on the process of myogenic differentiation needs to be explored further. Here, we show that the geometrical cues of substrates can significantly influence the differentiation process of C2C12 skeletal myoblasts. Three different geometries including lines of different widths, tori of different inner diameters, and hybrid structures (linear and circular features with different arc degrees) were created by micro-contact printing of fibronectin on the surface of Petri dishes. The differentiation of C2C12 cells was studied over a period of seven days and was quantified; we report the differentiation parameters of (1) fusion index, (2) degree of maturation, (3) alignment, and (4) response to electrical pulse stimulation (EPS). Hybrid structures with the smallest arc degree (hybrid 30°) showed the best results for all four differentiation parameters. The hybrid 30° pattern exhibits an ~2-fold increase in the fusion index when compared to the line patterns and an ~3-fold increase when compared to the toroid patterns. The hybrid 30° also showed a higher maturation index compared to the line or the toroid patterns. In response to electrical stimulation (20 V, 50 ms pulse, 1 Hz), mature myotubes on hybrid 30° patterns showed an ~2-fold increase in cellular displacement when compared to myotubes on the line and torus patterns. We tested the influence of C2C12 cell density on fusion and maturation indices, and the results suggest that density does not exert significant influence on cellular differentiation under these conditions. Our results can have important implications in engineering skeletal muscle tissues and designing muscle cell bio-actuators.


Cost-Effectiveness Analysis of Four Simulated Colorectal Cancer Screening Interventions, North Carolina.
Colorectal cancer (CRC) screening rates are suboptimal, particularly among the uninsured and the under-insured and among rural and African American populations. Little guidance is available for state-level decision makers to use to prioritize investment in evidence-based interventions to improve their population's health. The objective of this study was to demonstrate use of a simulation model that incorporates synthetic census data and claims-based statistical models to project screening behavior in North Carolina. We used individual-based modeling to simulate and compare intervention costs and results under 4 evidence-based and stakeholder-informed intervention scenarios for a 10-year intervention window, from January 1, 2014, through December 31, 2023. We compared the proportion of people living in North Carolina who were aged 50 to 75 years at some point during the window (that is, age-eligible for screening) who were up to date with CRC screening recommendations across intervention scenarios, both overall and among groups with documented disparities in receipt of screening. We estimated that the costs of the 4 intervention scenarios considered would range from $1.6 million to $3.75 million. Our model showed that mailed reminders for Medicaid enrollees, mass media campaigns targeting African Americans, and colonoscopy vouchers for the uninsured reduced disparities in receipt of screening by 2023, but produced only small increases in overall screening rates (0.2-0.5 percentage-point increases in the percentage of age-eligible adults who were up to date with CRC screening recommendations). Increased screenings ranged from 41,709 additional life-years up to date with screening for the voucher intervention to 145,821 for the mass media intervention. Reminders mailed to Medicaid enrollees and the mass media campaign for African Americans were the most cost-effective interventions, with costs per additional life-year up to date with screening of $25 or less. The intervention expanding the number of endoscopy facilities cost more than the other 3 interventions and was less effective in increasing CRC screening. Cost-effective CRC screening interventions targeting observed disparities are available, but substantial investment (more than $3.75 million) and additional approaches beyond those considered here are required to realize greater increases population-wide.


Educational Interventions for Teaching Evidence-Based Practice to Undergraduate Nursing Students: A Scoping Review.
Background: Evidence-based practice (EBP) is the appropriate approach to guide healthcare personnel in their clinical practice. Despite the importance of EBP, undergraduate nursing students are not very much engaged and have a lack of knowledge and skills. Aim: The aim of this study was to gather, assess and synthesize evidence on educational interventions promoting evidence-based practice competencies in traditional undergraduate nursing students. Methods: This is a scoping review on sixteen English and non-English databases. A data extraction form was established including authors, year of publication, country, types of participant, specific objectives, study design, educational intervention, comparison if existed, and outcomes of significance. Results: The search strategy retrieved 8901 records in total. After screening for duplicates and eligibility, 20 articles were included in the qualitative synthesis. Improvement in EBP domains such as knowledge, skills, attitudes/behaviours, EBP beliefs, use, practice, level of evidence, critical thinking and future use of EBP were mentioned and assessed in different studies. Conclusions: EBP training can improve nursing students' capacity in healthcare provision. Teaching EBP competencies along undergraduate nursing curricula should be a high priority at nursing programmes. The use of innovative approaches seems to be more effective than traditional ways. Education of EBP increases its future use and critical thinking and EBP programs improve self-efficacy and the level of evidence utilization.


A randomised controlled trial to compare clinical and cost-effectiveness of an online parent-led treatment for child anxiety problems with usual care in the context of COVID-19 delivered in Child and Adolescent Mental Health Services in the UK (Co-CAT): a study protocol for a randomised controlled trial.
In the context of COVID-19, NHS Child and Adolescent Mental Health Services (CAMHS) and other children's mental health services have faced major challenges in providing psychological treatments that (i) work when delivered remotely and (ii) can be delivered efficiently to manage increases in referrals as social distancing measures have been relaxed. Anxiety problems are a common reason for referral to CAMHS, children with pre-existing anxiety problems are particularly vulnerable in the context of COVID-19, and there were concerns about increases in childhood anxiety as schools reopened. The proposed research will evaluate the clinical and cost-effectiveness of a brief online parent-led cognitive behavioural treatment (CBT) delivered by the OSI (Online Support and Intervention for child anxiety) platform with remote support from a CAMHS therapist compared to 'COVID-19 treatment as usual' (C-TAU) in CAMHS and other children's mental health services throughout the COVID-19 pandemic. We will conduct a two-arm, multi-site, randomised controlled non-inferiority trial to evaluate the clinical and cost-effectiveness of OSI with therapist support compared to CAMHS and other child mental health services 'COVID-19 treatment as usual' (C-TAU) during the COVID-19 outbreak and to explore parent and therapists' experiences. If non-inferiority is shown, the research will provide (1) a solution for efficient psychological treatment for child anxiety disorders while social distancing (for the COVID-19 context and future pandemics); (2) an efficient means of treatment delivery as 'normal service' resumes to enable CAMHS to cope with the anticipated increase in referrals; and (3) a demonstration of rapid, high-quality evaluation and application of online interventions within NHS CAMHS to drive forward much-needed further digital innovation and evaluation in CAMHS settings. The primary beneficiaries will be children with anxiety disorders and their families, NHS CAMHS teams, and commissioners who will access a potentially effective, cost-effective, and efficient treatment for child anxiety problems. ISRCTN ISRCTN12890382 . Registered prospectively on 23 October 2020.


The medicinal chemist's toolbox for late stage functionalization of drug-like molecules.
The advent of modern C-H functionalization chemistries has enabled medicinal chemists to consider a synthetic strategy, late stage functionalization (LSF), which utilizes the C-H bonds of drug leads as points of diversification for generating new analogs. LSF approaches offer the promise of rapid exploration of structure activity relationships (SAR), the generation of oxidized metabolites, the blocking of metabolic hot spots and the preparation of biological probes. This review details a toolbox of intermolecular C-H functionalization chemistries with proven applicability to drug-like molecules, classified by regioselectivity patterns, and gives guidance on how to systematically develop LSF strategies using these patterns and other considerations. In addition, a number of examples illustrate how LSF approaches have been used to impact actual drug discovery and chemical biology efforts.


Charge carrier generation and transport in different stoichiometry APFO3:PC61BM solar cells.
In this paper we studied carrier drift dynamics in APFO3:PC61BM solar cells of varied stoichiometry (2:1, 1:1, and 1:4 APFO3:PC61BM) over a wide time range, from subpicoseconds to microseconds with a combination of ultrafast optical electric field probing and conventional transient integrated photocurrent techniques. Carrier drift and extraction dynamics are strongly stoichiometry dependent: the speed of electron or hole drift increases with higher concentration of PC61BM or polymer, respectively. The electron extraction from a sample with 80% PC61BM takes place during hundreds of picoseconds, but slows down to sub-microseconds in a sample with 33% PC61BM. The hole extraction is less stoichiometry dependent: it varies form sub-nanoseconds to tens of nanoseconds when the PC61BM concentration changes from 33% to 80%. The electron extraction rate correlates with the conversion efficiency of solar cells, leading to the conclusion that fast electron motion is essential for efficient charge carrier separation preventing their geminate recombination.


Microsatellite typing and avidity analysis suggest a common source of infection in herds with epidemic Neospora caninum-associated bovine abortion.
Neosporosis is an important cause of reproductive failure in cattle worldwide. Two different abortion patterns associated with Neospora caninum infection have been observed in cattle herds: endemic and epidemic abortion outbreaks. The endemic pattern is characterized by an abortion problem in a herd persisting for several months or years, and is assumed to be caused by reactivation of a chronic infection. In epidemic outbreaks, abortions concentrate within a short period of time, most likely due to a recent point source exposure of naïve animals to N. caninum. The aim of the study was to characterize five N. caninum-associated epidemic abortion outbreaks in Germany by serological and molecular techniques, including a p38-avidity-ELISA and typing of N. caninum in clinical samples by multilocus-microsatellite analysis. DNA extracts from the brain of 18 N. caninum infected fetuses from epidemic abortion outbreaks were characterized using 10 N. caninum-microsatellite markers. Nested-PCR protocols were developed to amplify the marker regions MS1B, MS3, MS5, MS6A, MS6B, MS7, MS12 and MS21 from clinical samples for subsequent analysis by capillary electrophoresis. Microsatellites MS2 and MS10 were analyzed by previously reported sequencing techniques. Most dams which had aborted showed a low-avidity IgG response to the N. caninum p38-antigen, and in three of the five studied herds, the majority of the dams at risk, which had not aborted, had also low-avidity responses suggesting that infection with N. caninum had recently occurred in most animals. A common microsatellite pattern prevailed in all fetuses from each individual epidemic outbreak. This pattern was unique for each herd. Although the number of epidemic abortion outbreaks analyzed was limited, the observation of a common microsatellite pattern, accompanied by a low-avidity IgG response against N. caninum in the dams, supports the hypothesis of a recent infection from a common point source. The genetic diversity of N. caninum observed among these outbreaks may indicate that not a particular N. caninum genotype but the horizontal infection route determines the occurrence of epidemic abortions.


Congregational participation and supports for children and adults with disabilities: parent perceptions.
Although religion and spirituality have received relatively limited attention in the literature addressing people with disabilities, each is strongly associated with a host of positive outcomes, including enhanced quality of life. One way to participate in religious activities and enhance spirituality is to participate in a faith community. In this article, we report findings from a survey of 416 parents exploring the ways in which they and their children with disabilities participated in their congregations and examining factors associated with participation and inclusion in those communities. Consistent with findings from general social surveys, the majority of parents indicated their faith was important to them and many-along with their sons or daughters with disabilities-participated in congregational activities. Although parents reported their sons and daughters with disabilities participated in somewhat fewer types of activities than they did, this involvement occurred most often in activities involving peers without disabilities. However, parents generally were not satisfied with the level of supports provided by faith communities, and they highly valued a welcoming and supportive attitude by the community. We discuss implications for extending inclusive efforts into congregational contexts and suggest future research directions.


A decorated raven bone from the Zaskalnaya VI (Kolosovskaya) Neanderthal site, Crimea.
We analyze a radius bone fragment of a raven (Corvus corax) from Zaskalnaya VI rock shelter, Crimea. The object bears seven notches and comes from an archaeological level attributed to a Micoquian industry dated to between 38 and 43 cal kyr BP. Our study aims to examine the degree of regularity and intentionality of this set of notches through their technological and morphometric analysis, complemented by comparative experimental work. Microscopic analysis of the notches indicate that they were produced by the to-and-fro movement of a lithic cutting edge and that two notches were added to fill in the gap left between previously cut notches, probably to increase the visual consistency of the pattern. Multivariate analysis of morphometric data recorded on the archaeological notches and sets of notches cut by nine modern experimenters on radii of domestic turkeys shows that the variations recorded on the Zaskalnaya set are comparable to experimental sets made with the aim of producing similar, parallel, equidistant notches. Identification of the Weber Fraction, the constant that accounts for error in human perception, for equidistant notches cut on bone rods and its application to the Zaskalnaya set of notches and thirty-six sets of notches incised on seventeen Upper Palaeolithic bone objects from seven sites indicate that the Zaskalnaya set falls within the range of variation of regularly spaced experimental and Upper Palaeolithic sets of notches. This suggests that even if the production of the notches may have had a utilitarian reason the notches were made with the goal of producing a visually consistent pattern. This object represents the first instance of a bird bone from a Neanderthal site bearing modifications that cannot be explained as the result of butchery activities and for which a symbolic argument can be built on direct rather than circumstantial evidence.


Elastic, dielectric and piezoelectric characterization of single domain PIN-PMN-PT: Mn crystals.
Mn modified 0.26Pb(In(1/2)Nb(1/2))O(3)-0.42Pb(Mg(1/3)Nb(2/3))O(3)-0.32PbTiO(3) (PIN-PMN-PT:Mn) single crystals with orthorhombic perovskite crystal structure were polarized along [011] direction, resulting in the single domain state "1O." The complete set of material constants was determined using the combined resonance and ultrasonic methods. The thickness shear piezoelectric coefficient d(15) and electromechanical coupling factor k(15) were found to be on the order of 3100 pC/N and 94%, respectively, much higher than longitudinal d(33) ∼ 270 pC/N and k(33) ∼ 70%. Using the single domain data, the rotated value of d(33)* along [001] direction was found to be 1230 pC/N, in agreement with the experimentally determined d(33) value of 1370 pC/N, conferring extrinsic contributions being about 10%, which was also confirmed using the Rayleigh analysis. In addition, the mechanical quality factors Q(m) were evaluated for different "1O" vibration modes, where the longitudinal Q(m) was found to be ∼1200, much higher than the value for "4O" crystals, ∼300.


Atrial fibrillation after transhiatal esophagectomy with transcervical endoscopic esophageal mobilization: one institution's experience.
There have been numerous studies regarding atrial fibrillation (AF) associated with cardiac and pulmonary surgery; however, studies looking at esophagectomy and atrial fibrillation are sparse. The goal of this study was to review our institution's atrial fibrillation rate following esophagectomy in order to better define the incidence and predisposing factors in this patient population. A retrospective chart review of all patients undergoing esophagectomy with transcervical endoscopic mobilization of the esophagus (TEEM) at the Medical College of Wisconsin and Affiliated Hospitals from July 2009 through December 2012. Seventy-one patients underwent TEEM esophagectomy during the study period. Of those, 23 (32.4%) patients developed new atrial fibrillation postoperatively. ICU (Intensive Care Unit) length of stay was 7.1 days for those that did not receive amiodarone, compared to 5.3 days for those that did receive amiodarone (p < 0.025). Those that went into AF spent on average 9.3 days in the ICU compared to 4.7 days for their counterparts that did not go into AF (p < 0.006). Total length of stay was not statistically different between populations [15.1 +/- 11.3 days compared to 13.5 +/- 9.4 days for those who did not go into AF (p < 0.281)]. Receiving preoperative amiodarone was found to reduce the overall incidence of AF. There was a trend towards decreased risk of going into AF in those who received preoperative amiodarone with an adjusted hazard ratio of 0.555 (p = 0.057). Similar to data reported in previous literature, postoperative atrial fibrillation was found to increase ICU length of stay as well as overall length of hospital stay. Preoperative amiodarone administration displayed a trend toward decreasing the rates of atrial fibrillation in patients undergoing TEEM.


Helping smokers quit: understanding the barriers to utilization of smoking cessation services.
Counseling smokers to quit smoking and providing them with pharmaceutical cessation aides are among the most beneficial and cost-effective interventions that clinicians can offer patients. Yet assistance with quitting is not universally covered by health plans or offered by all clinicians. Analysis of stakeholders' perspectives and interests can identify the barriers to more widespread provision of cessation services and suggest strategies for the public policy agenda to advance smoking cessation. Review of literature and discussions with representatives of stakeholders. All stakeholders-health plans, employers, clinicians, smokers, and the government-face barriers to broader smoking cessation activities. These range from health plans' perceiving that covering counseling and pharmacotherapy will increase costs without producing commensurate health care savings, to clinicians' feeling unprepared and uncompensated for counseling. Like other preventive measures aimed at behavior, efforts directed at smoking cessation have marginal status among health care interventions. State governments can help correct this status by increasing Medicaid coverage of treatment and expanding coverage for state employees. The federal government can promote the adoption of six initiatives recommended by a government subcommittee on cessation: set up a national quit line, develop a media campaign to encourage cessation, include cessation benefits in all federally funded insurance plans, create a research infrastructure to improve cessation rates, develop a clinician training agenda, and create a fund to increase cessation activities through a new $2 per pack cigarette excise tax. Both the federal and state governments can increase cessation by adopting policies such as the higher cigarette tax and laws prohibiting smoking in workplaces and public places. Public policy efforts should assume greater social responsibility for smoking cessation, including more aggressive leadership at the state and federal levels, as well as through advocacy, public health, and clinician organizations.


First Report of Phaeoacremonium scolyti Associated with Esca and Petri Diseases of Grapevine in Turkey.
Grapevine (Vitis vinifera L.) is a widely planted and economically important crop for production of raisin grapes, table grapes, and wine grapes in Turkey. Esca and petri diseases are two of the most important and destructive diseases of young and old vines worldwide (1). During the summers of 2009 and 2010, a survey was carried out in 63 vineyards in six locations of Ankara Province. Root and trunk samples were collected from 4- to 15-year-old grapevines showing esca and petri disease symptoms, including reduced trunk diameters, shortened internodes, stunted growth, chlorotic or necrotic leaves, and brown-to-black spots or streaks in the xylem vessels in cross or longitudinal sections of the rootstock trunk (1,3). Small pieces of internal tissues were surface disinfested in 1% sodium hypochlorite for 2 min, washed twice with sterile distilled water, and plated onto potato dextrose agar (PDA) amended with tetracycline hydrochloride (0.1 g liter-1). Plates were incubated at 25°C in the dark for 14 to 21 days. Phaeoacremonium spp. were consistently isolated from necrotic tissues. Single-conidial isolates of these Phaeoacremonium spp. were grown on PDA, malt extract agar (MEA), and oatmeal agar (OA) in the dark at 25°C for 2 to 3 weeks until colonies produced spores (3). Of these, Phaeoacremonium aleophilum was the most prevalent species, however, one isolate identified as P. scolyti was described by L. Mostert et al. (3). Conidiophores were mostly short and usually unbranched, subcylindrical to navicular, and often consisting of an elongate-ampuliform phialide. Phialides were terminal or lateral and pale brown to hyaline. Type II phialide were predominant. Type I phialide were 4 to 6 μm (average), type II phialide were 7 to 14 μm (average), and type III phialide were 14 to 20 μm (average). Conidia were hyaline, oblong-ellipsoidal, occasionally reniform or allantoid, 2.5 to 5 μm long (average), and 1 to 1.5 μm wide (average). Colony colors were reddish gray on PDA, pinkish white on MEA, and grayish pink on OA. Identity was confirmed by β-tubulin sequence analysis using primers T1 and Bt2b (2). Additionally, the β-tubulin gene fragment (primers T1 and Bt2b) of this isolate was sequenced (GenBank Accession No. JF909894). The sequence showed high similarity (99%) with the sequence of P. scolyti (GenBank Accession No. EU260415). Pathogenicity tests were completed using five, 3-month-old rooted cuttings of cv. Sultana. A hole approximately 3 mm in diameter was drilled on the crown 2 cm aboveground level from the bark to the pith and filled with a 30-μl spore suspension (106 spores/ml) harvested from 21-day-old cultures grown on PDA at 25°C in the dark. Five control plants were used. Controls were inoculated with sterile distilled water. Filled holes were sealed with a sheet of Parafilm. The plants were incubated for 3 months in a controlled environment facility at 25°C. After 3 months, the fungus was reisolated from black discoloration of vascular tissue and pith tissue of the crown area of all inoculated cuttings, completing Koch's postulates. The black discoloration was more compact near the point of inoculation. Control plants were asymptomatic and P. scolyti was not recovered from control plants. To our knowledge, this is the first report of the presence of P. scolyti causing esca and petri disease of grapevine in Turkey. References: (1) A. Eskalen et al. Plant Dis. 91:1100, 2007. (2) S. Essakhi et al. Persoonia 21:119, 2008. (3) L. Mostert et al. Stud. Mycol. 54:1, 2006.


Potential determining salts in microemulsions: interfacial distribution and effect on the phase behavior.
In this work we consider potential determining salts, also referred to as phase transfer agents for a future objective of electrochemistry at the oil-water interface in microemulsions. We have studied these salts, composed of a hydrophilic and a hydrophobic ion, in microemulsion stabilized by nonionic surfactants with an oligo ethylene oxide headgroup. NMR measurements show that the salts preferentially dissociate across the surfactant interface between the oil and water domains, and hence create a potential drop across the surfactant film, and back to back diffuse double layers in the oil and water phases. These observations are also supported by Poisson-Boltzmann calculations. This adsorption like event stabilizes the microemulsion. Repulsive long-range interactions between thin lamellae of surfactant and water lamellae in oil were observed using SAXS, thus confirming the presence of electrostatic forces mediated through the oil domain. We also observed that reversing the charges on the potential determining salts had opposite effects on the phase inversion temperature.


The influence of alcohol consumption on worldwide trends in mortality from upper aerodigestive tract cancers in men.
To assess current trends in male mortality from cancers of the oral cavity/pharynx, oesophagus, and larynx (upper aerodigestive tract cancers), and relate these to past national consumption of alcohol and smoking of cigarettes. To assess the impact of current trends in alcohol consumption and tobacco smoking on likely future rates of these cancers. Mortality data for cancers of the oral cavity/pharynx, oesophagus, and larynx were obtained for the years 1955-89 in 25 countries located in North America, Australasia, Europe, and Japan. Information on past and current alcohol consumption was also obtained for these countries, while current national lung cancer rates were used as a proxy measure of past smoking levels. The World Health Organization mortality database. National death rates from cancers of the oral cavity/pharynx, oesophagus, and larynx (considered together) are currently increasing among men and are most strongly associated with the level of per capita consumption of alcohol 20 years previously. They were less strongly associated with the level of alcohol consumption 10 years ago, and only very weakly associated with the current level of lung cancer mortality (a marker of past smoking habits). Regression analysis showed that the national rate of upper aerodigestive tract cancer could be estimated using information on past alcohol consumption and an interaction term between alcohol consumption and current lung cancer rates. Assuming stability in rates of lung cancer, the sizeable increase in alcohol consumption from 5 to 10 litres per capita each year that occurred in some countries during the 1960s and 70s means that increases of around 5 per 100,000 in the death rate from these cancers can be expected in these countries in the next decades. Previous alcohol consumption in a country is a strong predictor of deaths from cancers of the upper aerodigestive tract in men, and current increases in death rates can probably be related to increases in consumption which took place during the 1960s and 70s. Combined with a reduction in tobacco smoking, which is already taking place in some countries, reversing the trend of increases in consumption of alcohol has the potential for a sizeable impact on the burden of these cancers.


Evaluation of Prostate Cancer Bone Metastases with 18F-NaF and 18F-Fluorocholine PET/CT.
18F-fluorocholine is a specific promising agent for imaging tumor cell proliferation, particularly in prostate cancer, using PET/CT. It is a beneficial tool in the early detection of marrow-based metastases because it excludes distant metastases and evaluates the response to hormone therapy. In addition, 18F-fluorocholine has the potential to differentiate between degenerative and malignant osseous abnormalities because degenerative changes are not choline-avid; however, the agent may accumulate in recent traumatic bony lesions. On the other hand, 18F-NaF PET/CT can indicate increased bone turnover and is generally used in the assessment of primary and secondary osseous malignancies, the evaluation of response to treatment, and the clarification of abnormalities on other imaging modalities or clinical data. 18F-NaF PET/CT is a highly sensitive method in the evaluation of bone metastases from prostate cancer, but it has problematic specificity, mainly because of tracer accumulation in degenerative and inflammatory bone diseases. In summary, 18F-NaF PET/CT is a highly sensitive method, but 18F-fluorocholine PET/CT can detect early bone marrow metastases and provide greater specificity in the detection of bone metastases in patients with prostate cancer. However, the difference seems not to be significant.


Genetic structured antedependence and random regression models applied to the longitudinal feed conversion ratio in growing Large White pigs.
The objective of the present study was to compare a random regression model, usually used in genetic analyses of longitudinal data, with the structured antedependence (SAD) model to study the longitudinal feed conversion ratio (FCR) in growing Large White pigs and to propose criteria for animal selection when used for genetic evaluation. The study was based on data from 11,790 weekly FCR measures collected on 1,186 Large White male growing pigs. Random regression (RR) using orthogonal polynomial Legendre and SAD models was used to estimate genetic parameters and predict FCR-based EBV for each of the 10 wk of the test. The results demonstrated that the best SAD model (1 order of antedependence of degree 2 and a polynomial of degree 2 for the innovation variance for the genetic and permanent environmental effects, i.e., 12 parameters) provided a better fit for the data than RR with a quadratic function for the genetic and permanent environmental effects (13 parameters), with Bayesian information criteria values of -10,060 and -9,838, respectively. Heritabilities with the SAD model were higher than those of RR over the first 7 wk of the test. Genetic correlations between weeks were higher than 0.68 for short intervals between weeks and decreased to 0.08 for the SAD model and -0.39 for RR for the longest intervals. These differences in genetic parameters showed that, contrary to the RR approach, the SAD model does not suffer from border effect problems and can handle genetic correlations that tend to 0. Summarized breeding values were proposed for each approach as linear combinations of the individual weekly EBV weighted by the coefficients of the first or second eigenvector computed from the genetic covariance matrix of the additive genetic effects. These summarized breeding values isolated EBV trajectories over time, capturing either the average general value or the slope of the trajectory. Finally, applying the SAD model over a reduced period of time suggested that similar selection choices would result from the use of the records from the first 8 wk of the test. To conclude, the SAD model performed well for the genetic evaluation of longitudinal phenotypes.


Association of NRAMP1 polymorphism with leprosy type but not susceptibility to leprosy per se in west Africans.
Twin and family studies indicate that host genetic factors influence susceptibility to leprosy and, possibly, leprosy type. Murine studies have suggested a role for the natural resistance-associated macrophage protein 1 (Nramp1) gene, which can influence cellular immune responses to intracellular pathogens. We evaluated a variation in the human homolog, NRAMP1, recently associated with tuberculosis susceptibility in West Africa. A total of 273 patients with leprosy and 201 controls from Mali were genotyped for NRAMP1 polymorphisms previously associated with tuberculosis. No association was found with leprosy per se (P = 0.83), but the NRAMP1 3'-untranslated region 4-bp insertion/deletion polymorphism was associated with leprosy type (P = 0.007). Heterozygotes were more frequent among multibacillary than paucibacillary leprosy cases. Thus, variation in or near the NRAMP1 gene may exert an influence on the clinical presentation of leprosy, possibly by influencing cellular immune response type.


Cutaneous Lymphoma International Consortium Study of Outcome in Advanced Stages of Mycosis Fungoides and Sézary Syndrome: Effect of Specific Prognostic Markers on Survival and Development of a Prognostic Model.
Advanced-stage mycosis fungoides (MF; stage IIB to IV) and Sézary syndrome (SS) are aggressive lymphomas with a median survival of 1 to 5 years. Clinical management is stage based; however, there is wide range of outcome within stages. Published prognostic studies in MF/SS have been single-center trials. Because of the rarity of MF/SS, only a large collaboration would power a study to identify independent prognostic markers. Literature review identified the following 10 candidate markers: stage, age, sex, cutaneous histologic features of folliculotropism, CD30 positivity, proliferation index, large-cell transformation, WBC/lymphocyte count, serum lactate dehydrogenase, and identical T-cell clone in blood and skin. Data were collected at specialist centers on patients diagnosed with advanced-stage MF/SS from 2007. Each parameter recorded at diagnosis was tested against overall survival (OS). Staging data on 1,275 patients with advanced MF/SS from 29 international sites were included for survival analysis. The median OS was 63 months, with 2- and 5-year survival rates of 77% and 52%, respectively. The median OS for patients with stage IIB disease was 68 months, but patients diagnosed with stage III disease had slightly improved survival compared with patients with stage IIB, although patients diagnosed with stage IV disease had significantly worse survival (48 months for stage IVA and 33 months for stage IVB). Of the 10 variables tested, four (stage IV, age > 60 years, large-cell transformation, and increased lactate dehydrogenase) were independent prognostic markers for a worse survival. Combining these four factors in a prognostic index model identified the following three risk groups across stages with significantly different 5-year survival rates: low risk (68%), intermediate risk (44%), and high risk (28%). To our knowledge, this study includes the largest cohort of patients with advanced-stage MF/SS and identifies markers with independent prognostic value, which, used together in a prognostic index, may be useful to stratify advanced-stage patients.


Plasma fetuin-A concentration, genetic variation in the AHSG gene and risk of colorectal cancer.
Fetuin-A, also referred to as α2-Heremans-Schmid glycoprotein (AHSG), is a liver protein known to inhibit insulin actions. Hyperinsulinemia is a possible risk factor for colorectal cancer; however, the role of fetuin-A in the development of colorectal cancer is unclear. We investigated the association between circulating fetuin-A and colorectal cancer risk in a nested case-control study within the European Prospective Investigation into Cancer and Nutrition. Fetuin-A concentrations were measured in prediagnostic plasma samples from 1,367 colorectal cancer cases and 1,367 matched controls. In conditional logistic regression models adjusted for potential confounders, the estimated relative risk (95% confidence interval) of colorectal cancer per 40 µg/mL higher fetuin-A concentrations (approximately one standard deviation) was 1.13 (1.02-1.24) overall, 1.21 (1.05-1.39) in men, 1.06 (0.93-1.22) in women, 1.13 (1.00-1.27) for colon cancer and 1.12 (0.94-1.32) for rectal cancer. To improve causal inference in a Mendelian Randomization approach, five tagging single nucleotide polymorphisms of the AHSG gene were genotyped in a subset of 456 case-control pairs. The AHSG allele-score explained 21% of the interindividual variation in plasma fetuin-A concentrations. In instrumental variable analysis, genetically raised fetuin-A was not associated with colorectal cancer risk (relative risk per 40 µg/mL genetically determined higher fetuin-A was 0.98, 95% confidence interval: 0.73-1.33). The findings of our study indicate a modest linear association between fetuin-A concentrations and risk of colorectal cancer but suggest that fetuin-A may not be causally related to colorectal cancer development.


Not seeing the forest for the trees: size of the minimum spanning trees (MSTs) forest and branch significance in MST-based phylogenetic analysis.
Trees, including minimum spanning trees (MSTs), are commonly used in phylogenetic studies. But, for the research community, it may be unclear that the presented tree is just a hypothesis, chosen from among many possible alternatives. In this scenario, it is important to quantify our confidence in both the trees and the branches/edges included in such trees. In this paper, we address this problem for MSTs by introducing a new edge betweenness metric for undirected and weighted graphs. This spanning edge betweenness metric is defined as the fraction of equivalent MSTs where a given edge is present. The metric provides a per edge statistic that is similar to that of the bootstrap approach frequently used in phylogenetics to support the grouping of taxa. We provide methods for the exact computation of this metric based on the well known Kirchhoff's matrix tree theorem. Moreover, we implement and make available a module for the PHYLOViZ software and evaluate the proposed metric concerning both effectiveness and computational performance. Analysis of trees generated using multilocus sequence typing data (MLST) and the goeBURST algorithm revealed that the space of possible MSTs in real data sets is extremely large. Selection of the edge to be represented using bootstrap could lead to unreliable results since alternative edges are present in the same fraction of equivalent MSTs. The choice of the MST to be presented, results from criteria implemented in the algorithm that must be based in biologically plausible models.