Conservation of copper-transporting P(IB)-type ATPase function.
Copper-transporting P(IB)-type ATPases are highly conserved, and while unicellular eukaryotes and invertebrates have only one, a gene duplication has occurred during vertebrate evolution. Copper-induced trafficking of mammalian ATP7A and ATP7B from the trans-Golgi Network towards the plasma membrane is critical for their role in copper homeostasis. In polarized epithelial cells ATP7A and ATP7B traffic towards the basolateral and apical membranes respectively. We examined the localization and function of DmATP7, the single Drosophila melanogaster orthologue, in cultured D. melanogaster and mammalian cells to explore the conservation of P(IB)-type ATPase function. Comparative genomic analysis demonstrated motifs involved in basolateral targeting and retention of ATP7A were conserved in DmATP7, whereas ATP7B targeting motifs were not. DmATP7 expression was able to correct the copper hyper-accumulation phenotype of cultured fibroblasts from a Menkes disease patient expressing a null ATP7A allele. DmATP7 was able to transport copper to the cupro-enzyme tyrosinase and under elevated copper conditions DmATP7 was able to traffic towards the plasma membrane and efflux copper, essentially phenocopying ATP7A. When expressed in polarized Madin-Darby Canine Kidney cells, DmATP7 translocated towards the basolateral membrane when exposed to elevated copper, similar to ATP7A. These results demonstrate DmATP7 is able to functionally compensate for the absence of ATP7A, with important trafficking motifs conserved in these distantly related orthologues.


Reduced response to chronic mild stress in PACAP mutant mice is associated with blunted FosB expression in limbic forebrain and brainstem centers.
Pituitary adenylate cyclase-activating polypeptide (PACAP) has been implicated in stress adaptation with potential relevance in mood disorder management. PACAP deficient (KO) mice on CD1 background were shown to have depression-like phenotype. Here we aimed at investigating effects of chronic variable mild stress (CVMS) in non-injected, vehicle and imipramine-treated KO mice vs. wildtype (WT) counterparts. We hypothesized reduced FosB neuronal activity in stress-related centers, altered activity and peptide/neurotransmitter content of corticotropin-releasing factor (CRF) cells of the oval (ovBST) bed nucleus of stria terminalis (BST), urocortin 1 (Ucn1) neurons of centrally projecting Edinger-Westphal nucleus (cpEW) and serotonin (5HT) cells of dorsal raphe (DR) in PACAP deficiency. CVMS caused decreased body weight and increased adrenal size, corticosterone (CORT) titers and depression-like behavior in WT mice, in contrast to KO animals. CVMS increased FosB in the central (CeA) and medial amygdala, dorsomedial (dmBST), ventral (vBST), ovBST, CA1 area, dentate gyrus (DG), ventral lateral septum, parvo- (pPVN) and magnocellular paraventricular nucleus, lateral periaqueductal gray, cpEW and DR. Lack of PACAP blunted the CVMS-induced FosB rise in the CeA, ovBST, dmBST, vBST, CA1 area, pPVN and DR. The CVMS-induced FosB expression in ovBST-CRF and cpEW-Ucn1 neurons was abolished in KO mice. Although CVMS did not induce FosB in 5HT-DR neurons, PACAP KO mice had increased 5HT cell counts and 5HT content. We conclude that PACAP deficiency affects neuronal reactivity in a brain area-specific manner in stress centers, as well as in ovBST-CRF, cpEW-Ucn1 and 5HT-DR neurons leading to reduced CVMS response and altered depression level.


Investigating the over-representation of older persons in do-it-yourself home maintenance injury and barriers to prevention.
To examine why older persons undertake high-risk do-it-yourself (DIY) home maintenance and under what circumstances, what constitutes acceptable low-risk alternatives to DIY, and to assess if alternatives are feasible in the current context. Exploratory qualitative study using focus-group methodology. Fifteen focus groups were conducted, involving 118 persons aged 60 years and older, in two Melbourne communities. Participants resided locally, participated in local seniors groups, or received treatment for a DIY injury at one of two public hospitals serving these communities. Older persons' involvement in DIY ranged from necessity to choice. A number chose DIY for general fitness enhancement, satisfaction and pride in a job well done, and giving meaning and enjoyment to daily tasks. However, some older, frailer seniors were forced into DIY because of difficulties in choosing appropriate alternatives; lack of knowledge of some available resources and services; the challenge of accessing cost-effective and reliable private service providers; and fear of vulnerability to overcharging, overservicing or their personal security. Preferred DIY alternatives were local government providers, local paper advertised services, recommendations to private service providers and family, friends or neighbors. Lack of knowledge of other existing alternatives was an impediment to preventing DIY injury, or accessing DIY alternatives. A number of potentially feasible alternatives to DIY were identified from our review. This research is an important first step in understanding issues facing community-dwelling seniors remaining at home, and provides a basis on which government agencies and other providers can develop services to meet increasing needs.


Disintegrating force as a new formulation parameter.
Some coated aspirin tablet formulations were evaluated by relating their properties to disintegrating force development patterns. The treatment of disintegrating force-time curves was effected using the Weibull distribution as proposed for dissolution. Such parameters as the maximum disintegrating force developed, the time needed to reach 63.2% maximum disintegrating force (tau d) the shape parameter, the lag time, and the input value were used for evaluating the formulas examined. It was concluded that the input values, the integrating force development rate at tau d, can be employed as a new formulation parameter since, when correlated with the crushing strength, it allows an overall evaluation of the formula examined.


A kinetic and kinematic analysis of the effect of stochastic resonance electrical stimulation and knee sleeve during gait in osteoarthritis of the knee.
Extended use of knee sleeves in populations at risk for knee osteoarthritis progression has shown functional and quality of life benefits; however, additional comprehensive kinematic and kinetic analyses are needed to determine possible physical mechanisms of these benefits which may be due to the sleeve's ability to enhance knee proprioception. A novel means of extending these enhancements may be through stochastic resonance stimulation. Our goal was to determine whether the use of a knee sleeve alone or combined with stochastic resonance electrical stimulation improves knee mechanics in knee osteoarthritis. Gait kinetics and kinematics were assessed in subjects with medial knee osteoarthritis when presented with four conditions: control1, no electrical stimulation/sleeve, 75% threshold stimulation/sleeve, and control2. An increase in knee flexion angle throughout stance and a decrease in flexion moment occurring immediately after initial contact were seen in the stimulation/sleeve and sleeve alone conditions; however, these treatment conditions did not affect the knee adduction angle and internal knee abduction moment during weight acceptance. No differences were found between the sleeve alone and the stochastic resonance with sleeve conditions. A knee sleeve can improve sagittal-plane knee kinematics and kinetics, although adding the current configuration of stochastic resonance did not enhance these effects.


Does L-asparaginase influence efficacy or toxicity when added to a standard CHOP protocol for dogs with lymphoma?
The purpose of this study was to evaluate response rates, 1st remission duration (FRD), and toxicity in dogs with previously untreated lymphoma receiving an identical CHOP-based combination chemotherapy protocol with or without L-asparaginase (LASP). One hundred fifteen dogs with lymphoma were scheduled to receive an identical CHOP-based chemotherapy protocol that included L-ASP. However, because of manufacturer-imposed random rationing, 31 dogs did not receive L-ASP as scheduled. The 2 treatment groups were statistically similar with respect to signalment and presence of historical negative prognostic factors. No difference was observed in the median FRD whether dogs did or did not receive L-ASP (206 versus 217 days, respectively; P = .67). No difference was observed in the median overall survival times between dogs receiving or not receiving L-ASP (310 versus 308 days, respectively; P = .84). No statistical difference was observed with respect to overall response rate between dogs that did or did not receive L-ASP (89.3% versus 87.1%, respectively; P = .75). Complete response rates between the groups also were no different (83.3% and 77.4% for L-ASP and non-L-ASP groups, respectively; P = .59). Prevalence of toxicity (neutropenia, diarrhea, or vomiting) and treatment delays (P = .80) also were similar between groups. The results of this study suggest that exclusion of L-ASP in this multidrug protocol does not significantly impact outcome. Therefore, it may be more appropriate to reserve the use of L-ASP for treating relapse in dogs with lymphoma that have failed induction therapy.


Empagliflozin Enhances Autophagy, Mitochondrial Biogenesis, and Antioxidant Defense and Ameliorates Renal Ischemia/Reperfusion in Nondiabetic Rats.
Recent meta-analyses have shown that sodium-glucose cotransporter 2 (SGLT-2) inhibitors alleviate chronic kidney disease and acute kidney injury in diabetic patients. In this study, we aimed to investigate the effect of empagliflozin on renal ischemia/reperfusion (I/R) in nondiabetic rats and find the possible mechanisms. Experimental Approach. Eighteen male Wistar rats were randomly divided into three groups, including healthy control, ischemic control, and empagliflozin-treated group. Thirty minutes of bilateral renal ischemia was induced by clamping the renal hilum. Forty-eight hours after reopening the clamps, rats' blood samples and tissue specimens were collected. Empagliflozin 10 mg/kg was administered by gavage, 2 hours before ischemia and 24 hours after the first dose. I/R injury led to a significant rise in serum creatinine and blood urea nitrogen which was significantly decreased after treatment with empagliflozin. Empagliflozin also alleviated tubulointerstitial and glomerular damage and significantly decreased tissue histology scores. Empagliflozin decreased the increased levels of malondialdehyde, interleukin 1β, and tumor necrosis factor α. SGLT2 inhibition increased the decreased expression of nuclear factor erythroid 2-related factor 2 and PPARG coactivator 1 alpha that conduct antioxidant defense and mitochondrial biogenesis, respectively. Furthermore, empagliflozin markedly increased LC3-II/LC3-I and bcl2/bax ratios, showing its beneficial effect on activation of autophagy and inhibition of apoptosis. Despite its effects on diabetic nephropathy, empagliflozin did not activate the Sestrin2/AMP-activated protein kinase pathway in this study. Empagliflozin improved renal I/R injury in nondiabetic rats in this study by promoting autophagy and mitochondrial biogenesis and attenuation of oxidative stress, inflammation, and apoptosis.


Designed ankyrin repeat proteins as scaffolds for multivalent recognition.
Ankyrin repeat (AR) proteins are composed of tandem repeats of a basic structural motif of ca. 33 amino acid residues that form a β-turn followed by two antiparallel α-helices. Multiple repeats stack together in a modular fashion to form a scaffold that is ideally suited for the presentation of multiple functional groups and/or recognition elements. Here we describe a biosynthetic strategy that takes advantage of the modular nature of these proteins to generate multivalent ligands that are both chemically homogeneous and structurally well-defined. Glycosylated AR proteins cluster the tetrameric lectin concanavalin A (Con A) at a rate that is comparable to the rate of Con A aggregation mediated by globular protein conjugates and variable density linear polymers. Thus, AR proteins define a new class of multivalent ligand scaffolds that have significant potential application in the study and control of a variety of multivalent interactions.


Invasion and persistence of a selfish gene in the Cnidaria.
Homing endonuclease genes (HEGs) are superfluous, but are capable of invading populations that mix alleles by biasing their inheritance patterns through gene conversion. One model suggests that their long-term persistence is achieved through recurrent invasion. This circumvents evolutionary degeneration, but requires reasonable rates of transfer between species to maintain purifying selection. Although HEGs are found in a variety of microbes, we found the previous discovery of this type of selfish genetic element in the mitochondria of a sea anemone surprising. We surveyed 29 species of Cnidaria for the presence of the COXI HEG. Statistical analyses provided evidence for HEG invasion. We also found that 96 individuals of Metridium senile, from five different locations in the UK, had identical HEG sequences. This lack of sequence divergence illustrates the stable nature of Anthozoan mitochondria. Our data suggests this HEG conforms to the recurrent invasion model of evolution. Ordinarily such low rates of HEG transfer would likely be insufficient to enable major invasion. However, the slow rate of Anthozoan mitochondrial change lengthens greatly the time to HEG degeneration: this significantly extends the periodicity of the HEG life-cycle. We suggest that a combination of very low substitution rates and rare transfers facilitated metazoan HEG invasion.


Spiramycin: safety in man.
Spiramycin, a 16-membered lactone ring macrolide, has been in clinical use for the past 15 years with little serious associated toxicity. Gastrointestinal disturbance has usually been mild and no changes in gastrointestinal motility have been noted either experimentally or in humans, in contrast to other macrolides, such as erythromycin. Allergic reactions have been uncommon and mainly restricted to transient skin eruptions. Although liver injury is a possible complication of most macrolide treatments, no conclusive evidence for spiramycin-induced hepatitis is currently available, and, again in contrast to most other macrolides, the lack of drug interactions with spiramycin has been clearly established in biochemical, pharmacokinetic and clinical studies.


Tracheal stenosis.
Tracheal stenosis following assisted ventilation is becoming recognized as a major complication of tracheostomy, and the use of cuffed tracheostomy tubes. The possible causes and surgical management of this condition are discussed together with recommendations to minimize the possibility of post-tracheostomy tracheal stenosis.


Characterizing the Use of Exercise Testing in Repaired Tetralogy of Fallot Patients: A Multi-Institutional Survey.
Long-term survival for repaired Tetralogy of Fallot (rTOF) is excellent. We achieve this by close clinical monitoring to stratify prognosis and guide clinical decision-making. Cardiopulmonary exercise stress testing (CPET) is used to help guide clinical decision making; however, there are no clear guidelines for its use in this population. We sought to describe practice variability with regards to exercise testing for rTOF patients and how exercise data is used to guide management. We distributed a survey to pediatric cardiologists via email. Analyses were performed using qualitative statistics, two-sample T-tests, and chi-squared analysis. One-hundred and three clinicians completed the survey with 83% reporting that they routinely send symptomatic rTOF patients for exercise testing and 59% for asymptomatic patients. Respondents who routinely test asymptomatic patients reported higher levels of perceived helpfulness of exercise testing (p = 0.04) and comfort with CPET interpretation (p = 0.03). Nearly all respondents (92%) reported changing management primarily based on exercise testing results, with 62% reporting "occasionally changing management" and 10% reporting "frequently changing management". Results indicated that exercise test results influenced clinical decisions, such as the timing of interventions, need for additional imaging, or the initiation of exercise interventions. There was a statistically significant relationship between the perceived helpfulness of exercise testing and the likelihood of management changes (p < 0.01). The variability in attitudes and practices highlights the need for evidence-based guidelines addressing exercise testing in rTOF, particularly for asymptomatic patients.


Wear-pattern analysis in retrieved tibial inserts of mobile-bearing and fixed-bearing total knee prostheses.
Components from 73 failed knee replacements (TKRs) consisting of rotating-platform, mobile-bearing and fixed-bearing implants were examined to assess the patterns of wear. The patterns were divided into low-grade (burnishing, abrasion and cold flow) and high-grade (scratching, pitting/metal embedding and delamination) to assess the severity of the wear of polyethylene. The rotating-platform group had a higher incidence of low-grade wear on the upper surface compared with the fixed-bearing group. By contrast, high-grade wear comprising scratching, pitting and third-body embedding was seen on the lower surface. Linear regression analysis showed a significant correlation of the wear scores between the upper and lower surfaces of the tibial insert (R(2) = 0.29, p = 0.04) for the rotating-platform group, but no significant correlation was found for the fixed-bearing counterpart. This suggests that high-grade wear patterns on the upper surface are reduced with the rotating-platform design. However, the incidence of burnishing, pitting/third-body embedding and scratching wear patterns on the lower surface was higher compared with that in the fixed-bearing knee.


The Effect of Pulmonary Valve Replacement (PVR) Surgery on Hemodynamics of Patients Who Underwent Repair of Tetralogy of Fallot (TOF).
Pulmonary insufficiency (PI) frequently develops in patients who underwent repair of tetralogy of fallot (TOF). The aim of present study was to assess the effect of pulmonary valve replacement (PVR) on hemodynamics of patients who underwent repair of TOF. This retrospective cohort carried out between July 2010 and October 2012 among consecutive PVRs of 19 patients who underwent TOF surgery. The PVRs was performed using bioprosthetic (n=17) and mechanical (n=2) valves. Our data was collected during follow up visits within 6 to 12 month after PVR. Our results show that PVR significantly decreased right ventricular end-diastolic volume (180.89±13.78 vs. 107.21±12.02 ml/m(2), P < .01), right ventricular end-systolic volume (105.42±15.98 vs. 58.15±11.67 ml/m(2), P < .01), RV mass (47.78±6.20 vs. 30.68±8.95 g/m(2), P < .01), and PI (48.21±1.43% vs. 12.68±5.60%, P < .01). Moreover, left ventricular end-diastolic volume significantly increased (78.05±17.21 vs. 90.78±14.82 ml/m(2), P < .01) after PVR. The other hemodynamics indexes did not change, significantly. Despite the controversies about efficacy of PVR after repair of TOF, the remarkable improvement of hemodynamic is a supportive rationale for performing PVR surgery in TOF patients.


Modes of invasion of cancer of the larynx. A statistical, histological, and radioclinical analysis of 120 cases.
A statistical comparison between histologic and radioclinical data was performed on a series of 120 cases of total laryngectomies in order to study the modes of invasion of laryngeal cancer and to quantify the differences of results as evaluated by pathological and by clinical examinations. The first part of this study deals with the spread of cancer depending on its site of origin; the second part examines spread regardless of the primary location of the laryngeal tumor. The comparative results show that it was difficult to determine the deep invasion of structures such as the preepiglottic space and the cartilages from the radioclinical examination, especially in cases of cancer developed on a laryngocele. This study emphasizes the importance of the histological examination of laryngeal specimens as described here in order to codify the modes of invasion and, as a result, to help the otorhinolarynglologist in the choice of the best treatment.


Strategies to develop endogenous stem cell-recruiting bioactive materials for tissue repair and regeneration.
A leading strategy in tissue engineering is the design of biomimetic scaffolds that stimulate the body's repair mechanisms through the recruitment of endogenous stem cells to sites of injury. Approaches that employ the use of chemoattractant gradients to guide tissue regeneration without external cell sources are favored over traditional cell-based therapies that have limited potential for clinical translation. Following this concept, bioactive scaffolds can be engineered to provide a temporally and spatially controlled release of biological cues, with the possibility to mimic the complex signaling patterns of endogenous tissue regeneration. Another effective way to regulate stem cell activity is to leverage the inherent chemotactic properties of extracellular matrix (ECM)-based materials to build versatile cell-instructive platforms. This review introduces the concept of endogenous stem cell recruitment, and provides a comprehensive overview of the strategies available to achieve effective cardiovascular and bone tissue regeneration.


A review of cognitive training in schizophrenia.
Empirically supported treatments for schizophrenia now include a variety of psychosocial interventions, such as social skills training, vocational rehabilitation, and psychotherapy. As awareness of the functional importance of neurocognitive impairments in schizophrenia has increased, interest in treatments to improve cognition has grown. We review the literature on cognitive training (CT), which has been studied in 17 published randomized, controlled trials to date. The differential effectiveness of noncomputerized and computer-assisted interventions, with and without strategy coaching, and an environmental adaptation intervention, is examined. We conclude that the different types of approaches, whether computer assisted or not, all have effective components that hold promise for improving cognitive performance, symptoms, and everyday functioning. Our recommendations for further research, including the use of functional outcome measures and long-term followup, highlight the importance of improving ecological validity in this area of treatment research.


An Integrative Review of Postpartum Depression in Rural U.S. Communities.
This study's aim is to synthesize and summarize the literature on postpartum depression (PPD) in U.S. rural populations. Internationally, PPD has a high prevalence in rural communities. Although women in rural U.S. communities have higher rates of depression outside of the postpartum time period, little study has been conducted on PPD in U.S. rural populations. It is unknown whether rural women in the United States have high rates of PPD as is common in rural populations internationally. We used integrative literature review using Whittemore and Knafl's (2005) methodology. We searched the databases MEDLINE, CINAHL, PsycINFO, and Academic Search Premier with the words "postpartum depression" or "postnatal depression" and the word "rural." We found 11 articles with empirical data that met the criteria and thus were included in the review. Seven articles were quantitative, two were qualitative, one was mixed methods, and one was a nonexperimental design. Five foci emerged in the literature including (a) screening and prevalence, (b) demographic factors, (c) program creation and implementation, (d) mental health care seeking, and (e) social support. The results suggest that prevalence of PPD may be higher in rural U.S. areas, that implementing PPD prevention and screening programs for rural women is feasible, and that women in rural areas rely on informal networks and may face a stigma for seeking mental health care. Further research is needed on PPD in rural U.S. Specifically, this research should focus on the mix of variables found throughout this review such as race and income level. Nurses should lead for changes in clinical practice and policy that increase screening and interventions for PPD in rural communities.


Spontaneous Separation of Immiscible Organic Droplets on Asymmetric Wedge Channels with Hierarchical Microchannels.
Spontaneous separation of immiscible organic droplets has substantial research implications for environmental protection and resource regeneration. Compared to the widely explored separation of oil-water mixtures, there are fewer reports on separating mixed organic droplets on open surfaces due to the low surface tension differences. Efficient separation of mixed organic liquids by exploiting the rapid spontaneous transport of droplets on open surfaces remains a challenge. Here, through the fusion of inspiration from the fast droplet transport capability of Sarracenia trichome and the asymmetric wedge channel structure of shorebird beaks, this work proposes a spine with hierarchical microchannels and wedge channels (SHMW). Due to the synergistic effect of capillary force and asymmetric Laplace force, the SHMW can rapidly separate mixed organic droplets into two pure phases without requiring additional energy. In particular, the self-spreading of the oil solution on the open channel surface is utilized to amplify the surface energy difference between two droplets, and SHMW achieves the pickup of oil droplets floating on the surface of the organic solution. The maximum separation efficiency on 3-SHMW can reach 99.63%, and it can also realize the antigravity separation of mixed organic droplets with a surface tension difference as low as 0.87 mN·m-1. Furthermore, SHMW performs controllable separation, oil droplet pickup, and continuous separation and collection of mixed organic droplets. It is expected that this cooperative structure composed of hierarchical microchannels and wedge channels will be realized in resource recovery or chemical reactions in industrial production processes.


Altered expression pattern of integrin alphavbeta3 correlates with actin cytoskeleton in primary cultures of human breast cancer.
Integrins are transmembrane adhesion receptors that provide the physical link between the actin cytoskeleton and the extracellular matrix. It has been well established that integrins play a major role in various cancer stages, such as tumor growth, progression, invasion and metastasis. In breast cancer, integrin alphavbeta3 has been associated with high malignant potential in cancer cells, signaling the onset of widespread metastasis. Many preclinical breast cancer studies are based on established cell lines, which may not represent the cell behavior and phenotype of the primary tumor of origin, due to undergone genotypic and phenotypic changes. In the present study, short-term primary breast cancer cell cultures were developed. Integrin alphavbeta3 localization was studied in correlation with F-actin cytoskeleton by means of immunofluorescence and immunogold ultrastructural localization. Integrin fluorescence intensities were semi-quantitatively assessed by means of computerized image analysis, while integrin and actin expression was evaluated by Western immunoblotting. In the primary breast cancer epithelial cells integrin alphavbeta3 immunofluorescence was observed in the marginal cytoplasmic area, whereas in the primary normal breast epithelial cells it was observed in the main cell body, i.e. in the ventrally located perinuclear area. In the former, F-actin cytoskeleton appeared well-formed, consisting of numerous and thicker stress fibers, compared to normal epithelial cells. Furthermore, electron microscopy showed increased integrin alphavbeta3 immunogold localization in epithelial breast cancer cells over the area of stress fibers at the basal cell surface. These findings were verified with Western immunoblotting by the higher expression of integrin beta3 subunit and actin in primary breast cancer cells, revealing their reciprocal relation, in response to the higher motility requirements, determined by the malignant potential of the breast cancer cells. A model system of primary breast cancer cell cultures was developed, in an effort to maintain the closest resembling environment to the tumor of origin. Using the above system model as an experimental tool the study of breast tumor cell behavior is possible concerning the adhesion capacity and the migrating potential of these cells, as defined by the integrin alphavbeta3 distribution in correlation with F-actin cytoskeleton.


[Who dies of morphine and dextropropoxyphene intoxication? Danish experiences from the period 1979-1992].
We studied deaths following intoxication with dextropropoxyphene (D) and opioids (M) in Denmark 1979-1992 with special reference to the sex, age group, contributory cause of death (secondary diagnosis) and manner of death. Deaths following D increased until 1985 for both sexes, where a total of 46 women and 64 men died. In 1985 the National Board of Health drew public attention to this problem which led to a decrease in these deaths among men, while in women a paradoxical increase in suicides outnumbered a reduction in deaths from intoxication accidents. In 1988 D was assigned to the more restrictive prescription rules of opioids, which further reduced the number of deaths. The reduction of D deaths was followed by a corresponding increase in deaths due to M. However, the demographic characteristics of D and M deaths were not entirely identical: The typical D victim had a history of psychiatric disease or drug/alcohol abuse and committed suicide; the age was 40-59 for women and 20-39 for men. The typical M victim also had a history of psychiatric disease and substance abuse but suicides were less common and the majority occurred in the age group 20-39 in both sexes. Both D and M deaths were rare in persons with a somatic secondary diagnosis. We conclude that these poisonings warrant continued attention, and that a more restrictive prescription practice of D and M to patients at risk is justified.


One-stage Posterior Approach in the Treatment of Consecutive Multi-segment Thoracic Tuberculosis with Kyphosis.
To evaluate the clinical efficacy of the one-stage posterior approach in treating consecutive multiple-segment thoracic tuberculosis with kyphosis. A retrospective study was performed. The data of 22 patients with consecutive multi-level thoracic tuberculosis and kyphosis who underwent the one-stage posterior approach of debridement, bone grafting, drainage and pedicle screws fixation were collected. The kyphotic angle was 61.6°±3.1° pre-operatively. The neurologic status based on American Spinal Injury Association (ASIA) scoring was A in 5 cases, B in 4 cases, C in 9 cases, D in 3 cases and E in 1 case. The average follow-up period was 43.4±5.0 months. The kyphotic angle was 19.8°±0.7° after operation (p < 0.01), and 21.9°±1.2° at the last visit (p < 0.01). No neurological deterioration was observed in any patients after surgery. A significant difference in ASIA score was detected when the state before the operation and at the last visit was compared (p < 0.01). There was no recurrent paravertebral abscess of thoracic tuberculosis in any of these cases. The one-stage posterior approach of debridement and drainage is a feasible surgical option for consecutive multisegment thoracic tuberculosis with kyphosis. It can achieve debridement, decompression, and stabilization simultaneously.


Power Profiles of Competitive and Noncompetitive Mountain Bikers.
Novak, AR, Bennett, KJM, Pluss, MA, Fransen, J, Watsford, ML, and Dascombe, BJ. Power profiles of competitive and noncompetitive mountain bikers. J Strength Cond Res 33(2): 538-543, 2019-The performance of Olympic distance cross-country mountain bikers (XCO-MTB) is affected by constraints such as erosion of track surfaces and mass start congestion which can affect race results. Standardized laboratory assessments quantify interseasonal and intraseasonal cycling potential through the assessment of multiple physiological capacities. Therefore, this study examined whether the power profile assessment (PPA) could discriminate between competitive XCO-MTB and noncompetitive mountain bikers (NC-MTB). Second, it aimed to report normative power profile data for competitive XCO-MTB cyclists. Twenty-nine male participants were recruited across groups of XCO-MTB (n = 14) and NC-MTB (n = 15) mountain bikers. Each cyclist completed a PPA that consisted of increasing duration maximal efforts (6, 15, 30, 60, 240, and 600 seconds) that were interspersed by longer rest periods (174, 225, 330, 480, and 600 seconds) between efforts. Normative power outputs were established for XCO-MTB cyclists ranging between 13.8 ± 1.5 W·kg (5-second effort) and 4.1 ± 0.6 W·kg (600-second effort). No differences in absolute peak power or cadence were identified between groups across any effort length (p > 0.05). However, the XCO-MTB cyclists produced greater mean power outputs relative to body mass than the NC-MTB during the 60-second (6.9 ± 0.8 vs 6.4 ± 0.6 W·kg; p = 0.002), 240-second (4.7 ± 0.7 vs. 3.8 ± 0.4 W·kg; p < 0.001), and 600-second (4.1 ± 0.6 vs. 3.4 ± 0.3 W·kg; p < 0.001) efforts. The PPA is a useful discriminative assessment tool for XCO-MTB and highlights the importance of aerobic power for XCO-MTB performance.


[Periodic movements of the limbs during sleep in childhood].
Periodic movements of the limbs (PML) whilst sleeping is a disorder included in the group of intrinsic dysomnias which cause problems of insomnia and somnolence during the day. It is usually associated with the restless legs syndrome (it occurs in 80% of the patients with this syndrome) although PML may occasionally be an isolated finding. The prevalence of the disease is of 11%. It usually occurs in patients aged over 50 years, and the frequency increase with age. It is uncommon in persons aged under 30 years. In children it is almost unknown. When it does occur in this age group it is usually associated with the restless legs syndrome. To obtain data regarding this disorder in childhood, in view of the scarcity of information available in the literature and the importance of the neuropsychological problems and difficulty with learning that affected children have. We report two cases of PML occurring in girls of under 7 years of age. They had severe associated neuropsychological disorders (disorders of behavior and feeding, psychotic episodes, etc.). The effect on performance at school led to complete failure in one case. We describe the clinical course of the two girls, emphasizing the variability, and both diagnostic and therapeutic difficulties. Although this condition is uncommon in childhood, it should always be borne in mind in cases of children with sleep disorders, since early diagnosis will prevent major behavior and learning problems.


Cerebral hypervascularity and early venous opacification in status epilepticus. A case report.
Cerebral hyperperfusion and early draining veins directly resulting from status epilepticus can be demonstrated angiographically. Postmortem examination of the brain excluded other possible causes of hyperemia and early venous drainage.


The protein kinase MAP3K19 phosphorylates MAP2Ks and thereby activates ERK and JNK kinases and increases viability of KRAS-mutant lung cancer cells.
Identifying additional mitogen-activated protein kinase (MAPK) pathway regulators is invaluable in aiding our understanding of the complex signaling networks that regulate cellular processes, including cell proliferation and survival. Here, using in vitro kinase assays and by expressing WT or kinase-dead MAPK kinase kinase 19 (MAP3K19) in the HEK293T cell line and assessing activation of the extracellular signal-regulated kinase (ERK) and JUN N-terminal kinase (JNK) signaling pathways, we defined MAP3K19 as a novel regulator of MAPK signaling. We also observed that overexpression of WT MAP3K19 activates both the ERK and JNK pathways in a panel of cancer cell lines. Furthermore, MAP3K19 sustained ERK pathway activation in the presence of inhibitors targeting the RAF proto-oncogene Ser/Thr protein kinase (RAF) and MAPK/ERK kinase, indicating that MAP3K19 activates ERK via a RAF-independent mechanism. Findings from in vitro and in-cell kinase assays demonstrate that MAP3K19 is a kinase that directly phosphorylates both MAPK/ERK kinase (MEK) and MAPK kinase 7 (MKK7). Results from an short-hairpin RNA screen indicated that MAP3K19 is essential for maintaining survival in KRAS-mutant cancers; therefore, we depleted or inhibited MAP3K19 in KRAS-mutant cancer cell lines and observed that this reduces viability and decreases ERK and JNK pathway activation. In summary, our results reveal that MAP3K19 directly activates the ERK and JNK cascades and highlight a role for this kinase in maintaining survival of KRAS-mutant lung cancer cells.


Failure of TGF beta1 and IL-12 to regulate human FasL and mTNF alloreactive cytotoxic T-cell pathways.
The effect of TGFbeta1 and IL-12 on calcium-independent cytotoxic pathways was investigated. We have previously demonstrated that the regulatory effect of TGFbeta1 and IL-12 on human alloreative CTL activity was associated with regulation of perforin and granzyme B gene expression. To determine the effect of both cytokines on the alternative cytotoxic pathway involving FasL and mTNF, we first investigated the expression of both molecules on human primary alloactivated T cells. Our results show that human allostimulated T lymphocytes express FasL. Cell lysis experiments demonstrate that the FasL cytotoxic pathway is involved in the killing of specific target cells mediated by human alloreactive CTL. In addition, allogeneic stimulation induced significant mTNF expression on both CD4+ and CD8+ responder T cells. Using TNF-sensitive target cells, we also demonstrate that the mTNF-mediated cytotoxic pathway is involved in the cytotoxic activity of human primary allostimulated T lymphocytes. Neither TGFbeta1 nor IL-12 had an effect on FasL or mTNF expression. Furthermore, addition of TGFbeta1 or IL-12 at the initiation of the MLR had no significant effect on Fas- and mTNF-mediated cytotoxicity. Taken together, our results provide a novel insight into the differences between regulation by cytokines of perforin-dependent and -independent cytotoxic mechanisms. Unlike their role in the perforin/granzyme B pathway, TGFbeta1 and IL-12 do not appear to mediate any regulatory effect on FasL and mTNF cytotoxic pathways used by human alloreactive primary CTL.


Proton FLASH Radiotherapy for the Treatment of Symptomatic Bone Metastases: The FAST-01 Nonrandomized Trial.
To our knowledge, there have been no clinical trials of ultra-high-dose-rate radiotherapy delivered at more than 40 Gy/sec, known as FLASH therapy, nor first-in-human use of proton FLASH. To assess the clinical workflow feasibility and treatment-related toxic effects of FLASH and pain relief at the treatment sites. In the FAST-01 nonrandomized trial, participants treated at Cincinnati Children's/UC Health Proton Therapy Center underwent palliative FLASH radiotherapy to extremity bone metastases. Patients 18 years and older with 1 to 3 painful extremity bone metastases and life expectancies of 2 months or more were eligible. Patients were excluded if they had foot, hand, and wrist metastases; metastases locally treated in the 2 weeks prior; metal implants in the treatment field; known enhanced tissue radiosensitivity; and implanted devices at risk of malfunction with radiotherapy. One of 11 patients who consented was excluded based on eligibility. The end points were evaluated at 3 months posttreatment, and patients were followed up through death or loss to follow-up for toxic effects and pain assessments. Of the 10 included patients, 2 died after the 2-month follow-up but before the 3-month follow-up; 8 participants completed the 3-month evaluation. Data were collected from November 3, 2020, to January 28, 2022, and analyzed from January 28, 2022, to September 1, 2022. Bone metastases were treated on a FLASH-enabled (≥40 Gy/sec) proton radiotherapy system using a single-transmission proton beam. This is consistent with standard of care using the same prescription (8 Gy in a single fraction) but on a conventional-dose-rate (approximately 0.03 Gy/sec) photon radiotherapy system. Main outcomes included patient time on the treatment couch, device-related treatment delays, adverse events related to FLASH, patient-reported pain scores, and analgesic use. A total of 10 patients (age range, 27-81 years [median age, 63 years]; 5 [50%] male) underwent FLASH radiotherapy at 12 metastatic sites. There were no FLASH-related technical issues or delays. The average (range) time on the treatment couch was 18.9 (11-33) minutes per patient and 15.8 (11-22) minutes per treatment site. Median (range) follow-up was 4.8 (2.3-13.0) months. Adverse events were mild and consistent with conventional radiotherapy. Transient pain flares occurred in 4 of the 12 treated sites (33%). In 8 of the 12 sites (67%) patients reported pain relief, and in 6 of the 12 sites (50%) patients reported a complete response (no pain). In this nonrandomized trial, clinical workflow metrics, treatment efficacy, and safety data demonstrated that ultra-high-dose-rate proton FLASH radiotherapy was clinically feasible. The treatment efficacy and the profile of adverse events were comparable with those of standard-of-care radiotherapy. These findings support the further exploration of FLASH radiotherapy in patients with cancer. ClinicalTrials.gov Identifier: NCT04592887.


Detection of unsuspected central pulmonary embolism with conventional contrast-enhanced CT.
When fast, nondynamic scanning techniques are used in combination with adequate intravenous injection of contrast material, good-quality images of the pulmonary artery (PA) can be obtained with routine computed tomography (CT). The purpose of this study was to evaluate PA filling defects detected on these CT scans. Twenty-two CT scans from 22 patients with PA filling defects were reviewed. A routine scanning technique was used. Thrombus was unilateral in 12 cases and bilateral in 10. All patients but one had a predisposition for pulmonary embolism (PE). However, only five patients underwent CT because there was a strong suspicion for PE. In 17 cases, PE was not the first choice of clinical diagnosis; in 13 cases, thrombus of the PA was not even considered. Diagnosis was confirmed in 11 patients with ventilation-perfusion scanning (n = 7), angiography (n = 3), and surgery (n = 1). In 11 patients, anticoagulation therapy was started without further diagnostic procedures. Diagnosis of thrombus of the PA is possible with routine nondynamic, contrast material-enhanced CT. Because PE is often unsuspected at clinical examination, every contrast-enhanced CT scan of the chest should be evaluated for the presence of filling defects in the PA.


Effect of A23187 ionophore treatment on human blastocyst development-a sibling oocyte study.
To investigate whether treatment with commercially available ready-to-use A23187 ionophore (GM508-CultActive) improves embryo development outcome in patients with a history of embryo developmental problems. This is a uni-center prospective study in which sibling oocytes of patients with embryos of poor quality on day 5 in the previous cycle were treated or not with CultActive. Two hundred forty-seven metaphase II (MII) oocytes from 19 cycles performed between 2016 and 2019 were included in the study. After ICSI, the sibling oocytes were assigned to the treatment group or to the control group, following an electronically generated randomization list. A number of 122 MII were treated with CultActive and 125 MII had no treatment and were assigned to the control group. No difference in fertilization rate (p = 0.255) or in the capacity of embryos to reach good quality on day 5 (p = 0.197) was observed between the two groups. The utilization rates defined as the number of embryos transferred or cryopreserved per mature oocyte (p = 0.438) or per fertilized oocytes (p = 0.299) were not significantly different between the treated group and the control group. The results of the current study do not support the use of CultActive in cases with embryo developmental problems.


A pilot study examining patient attitudes and intentions to adopt assistive technologies into type 2 diabetes self-management.
Approximately half of individuals living with type 2 diabetes mellitus (T2DM) have suboptimal self-management, which could be improved by using assistive technologies in self-management regimes. This study examines patient attitudes and intentions to adopt assistive technologies into T2DM self-management. Forty-four participants (M = 58.7 years) with T2DM were recruited from diabetes education classes in the southwestern Ontario, Canada, between February and April 2014. Participants completed a self-reported in-person survey assessing demographic characteristics, current diabetes management, and attitudes toward using assistive technologies in their diabetes self-management. Demographics, disease characteristics, and current technology use and preferences of the cohort were examined, followed by a correlational analysis of descriptive characteristics and attitudes and intentions to use technology in self-management. The majority of (but not all) participants felt that using Internet applications (65%) and smartphone (53.5%) applications for self-management was a good idea. The majority of participants did not currently use an Internet (92.5%) or mobile (96%) application for self-management. Of participants, 77% intended to use an Internet application to manage their diabetes in the future and 58% intended to use mobile applications. Younger age was associated with more positive attitudes (r = -.432, P = .003) and intentions (r = -.425, P = .005) to use assistive technologies in diabetes self-management. Findings suggest that patients, especially those younger in age, are favorable toward adopting assistive technologies into management practice. However, attitudes among older adults are less positive, and few currently make use of such technologies in any age group.


Doctoring by go-between: aspects of health care for Malay children.
The paper is a preliminary account of the way Malay adults handle sickness episodes when they occur among their children. We first show how children are conceived of as different from adults, then give some of the reasons for their vulnerability: their 'vital principle' is weak and unstable, they react more violently to humoral variations, they are related to and attracted by the supernatural world. The second section gives a sample of common indigenous illness categories as applied to children, then summarizes a typical aetiological model including factors that are children-specific. Such factors often point to either parent as initiating the episode by disrupting the animal, symbolic or social world, or by placing the child out of harmony with the outside world. The third section provides an account of therapeutic alternatives, some of which are coherent with the aetiology, others--aiming at restoring the whole person's balance--are not specific of a particular sickness. Both types can be provided by native curers, whereas only the former is expected from urban based general practitioners; we show how Malay parents in fact partly relate to medical doctors as if they were traditional healers. To sum up, we emphasize the mediating role of adults towards sick children: not only do they provide identification and treatment but they often hold themselves responsible for the disorder. We also show the socializing, hence integrating, function of sickness for children: they are gradually taught about the world order, and introduced to the value system of the culture in which they are growing up. We also note that both aetiologies and therapies can be children-specific.(ABSTRACT TRUNCATED AT 250 WORDS)


Dual Strategy with Adipose-derived Stem Cells and L-arginine Recovered Cavernosal Functions in a Rat Model of Radical Prostatectomy.
As standard therapy for prostate cancer, radical prostatectomy (RP) causes cavernous nerve injury and increases fibrosis and hypoxia-induced penile structural alterations. This study aimed to determine the potential beneficial effects of adipose-derived stem cells (ADSCs) and L-arginine alone or in combination on the penile erection of a rat model of erectile dysfunction (ED) caused by bilateral cavernous nerve (CN) transection. Male rats (n=35) were randomized into five groups: Sham-operated; CN transection (4-wk); CN transection plus ADSCs (1x106 cells by intracavernosal injection); CN transection plus L-arginine (4wk, 10 mg/kg/day, oral); and ADSCs combined with L-arginine in CN transection. In vivo erectile responses and in vitro relaxant responses were measured. Western blot and immunohistochemistry analyses were used to determine the expression and localization of endothelial nitric oxide synthases (eNOS), neuronal NOS (nNOS), transforming growth factor-beta 1 (TGF-1), hypoxia-inducible factor-1 (HIF-1), and apoptosis markers (Bax and Bcl-2). The ratio of smooth muscle to collagen and nerve regeneration was calculated using Masson's trichrome and nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase staining. The combined treatment restored diminished erectile responses, endothelium-dependent acetylcholine, and electrical field stimulation-induced relaxation of the corpus cavernosum in rats with CN transection, whereas either monotherapy produced only partial improvements. All treatment regimens restored increases in the protein expression of HIF-1 and Bax in rodents with CN transection. The decrease in smooth muscle mass and NADPH-diaphorase-positive nerve fibers was partially ameliorated by monotherapy, whereas combined therapy led to recovery. These findings indicate that combined treatment with ADSCs and L-arginine may restore erectile function in rodents with CN transection by inhibiting hypoxia-induced neurotoxicity and preserving endothelium function and smooth muscle content.


EXPLORING PERSONALITY DIAGNOSIS STABILITY FOLLOWING ACUTE PSYCHOTHERAPY FOR CHRONIC POSTTRAUMATIC STRESS DISORDER.
Axis I comorbidity complicates diagnosing axis II personality disorders (PDs). PDs might influence Axis I outcome. No research has examined psychotherapy effects on PDs of treating Axis I comorbidity. Secondary analysis of a randomized controlled trial examined PD diagnostic stability after brief psychotherapy of chronic posttraumatic stress disorder (PTSD). Patients with chronic PTSD were randomly assigned to 14 weeks of prolonged exposure, interpersonal psychotherapy, or relaxation therapy. Assessments included the Structured Clinical Interview for DSM-IV, Patient Version (SCID-P) and Structured Clinical Interview for DSM-IV Axis II Disorders (SCID-II) at baseline, week 14, and for treatment responders (≥30% clinician-administered PTSD scale improvement, defined a priori) at week 26 follow-up. We hypothesized patients whose PTSD improved would retain fewer baseline PD diagnoses posttreatment, particularly with personality traits PTSD mimics, e.g. paranoid and avoidant. Forty-seven (47%) of 99 SCID-II patients evaluated at baseline received a SCID-II diagnosis: paranoid (28%), obsessive-compulsive (27%), and avoidant (23%) PDs were most prevalent. Among 78 patients who repeated SCID-II evaluations posttreatment, 45% (N = 35) had baseline PD diagnoses, of which 43% (N = 15/35) lost at week 14. Three (7%) patients without baseline PDs acquired diagnoses at week 14; 10 others shifted diagnoses. Treatment modality and PTSD response were unrelated to PD improvement. Of treatment responders reevaluated at follow-up (N = 44), 56% with any baseline Axis II diagnosis had none at week 26. This first evaluation of Axis I psychotherapy effects on personality disorder stability found that acutely treating a chronic state decreased apparent trait-across most PDs observed. These exploratory findings suggest personality diagnoses may have limited prognostic meaning in treating chronic PTSD.


Genetic variations and miRNA-target interactions contribute to natural phenotypic variations in Populus.
Variation in regulatory factors, including microRNAs (miRNAs), contributes to variation in quantitative and complex traits. However, in plants, variants in miRNAs and their target genes that contribute to natural phenotypic variation, and the underlying regulatory networks, remain poorly characterized. We investigated the associations and interactions of single-nucleotide polymorphisms (SNPs) in miRNAs and their target genes with phenotypes in 435 individuals from a natural population of Populus. We used RNA-seq to identify 217 miRNAs differentially expressed in a tension wood system, and identified 1196 candidate target genes; degradome sequencing confirmed 60 of the target sites. In addition, 72 miRNA-target pairs showed significant co-expression. Gene ontology (GO) term analysis showed that most of the genes in the co-regulated pairs participate in biological regulation. Genome resequencing found 5383 common SNPs (frequency ≥ 0.05) in 139 miRNAs and 31 037 SNPs in 819 target genes. Single-SNP association analyses identified 232 significant associations between wood traits (P ≤ 0.05) and SNPs in 102 miRNAs and 1387 associations with 478 target genes. Among these, 102 miRNA-target pairs associated with the same traits. Multi-SNP associations found 102 epistatic pairs associated with traits. Furthermore, a reconstructed regulatory network contained 12 significantly co-expressed pairs, including eight miRNAs and nine targets associated with traits. Lastly, both expression and genetic association showed that miR156i, miR156j, miR396a and miR6445b were involved in the formation of tension wood. This study shows that variants in miRNAs and target genes contribute to natural phenotypic variation and annotated roles and interactions of miRNAs and their target genes by genetic association analysis.


Multiple breath washout in pediatric patients after lung transplantation.
Forced expiratory volume in 1 second (FEV1 ) from spirometry is the most commonly used parameter to detect early allograft dysfunction after lung transplantation (LTx). There are concerns regarding its sensitivity. Nitrogen-multiple breath washout (N2 -MBW) is sensitive at detecting early global (lung clearance index [LCI]) and acinar (Sacin ) airway inhomogeneity. We investigated whether N2 -MBW indices indicate small airways pathology after LTx in children with stable spirometry. Thirty-seven children without bronchiolitis obliterans syndrome [BOS] at a median of 1.6 (0.6-3.0) years after LTx underwent N2 -MBW and spirometry, 28 of those on 2 occasions (≤6 months apart) during clinically stable periods. Additional longitudinal data (11 and 8 measurements, respectively) are provided from 2 patients with BOS. In patients without BOS, LCI and Sacin were significantly elevated compared with healthy controls. LCI was abnormal at the 2 test occasions in 81% and 71% of patients, respectively, compared with 30% and 39% of patients with abnormal FEV1 /forced vital capacity (FVC). Correlations of LCI with FEV1 /FVC (r = 0.1, P = .4) and FEV1 (r = -0.1, P = .6) were poor. N2 -MBW represents a sensitive and reproducible tool for the early detection of airways pathology in stable transplant recipients. Moreover, indices were highly elevated in both patients with BOS. Spirometry and LCI showed poor correlation, indicating distinct and complementary physiologic measures.


Podocyte as the target for aldosterone: roles of oxidative stress and Sgk1.
Accumulating evidence suggests that mineralocorticoid receptor blockade effectively reduces proteinuria in hypertensive patients. However, the mechanism of the antiproteinuric effect remains elusive. In this study, we investigated the effects of aldosterone on podocyte, a key player of the glomerular filtration barrier. Uninephrectomized rats were continuously infused with aldosterone and fed a high-salt diet. Aldosterone induced proteinuria progressively, associated with blood pressure elevation. Notably, gene expressions of podocyte-associated molecules nephrin and podocin were markedly decreased in aldosterone-infused rats at 2 weeks, with a gradual decrease thereafter. Immunohistochemical studies and electron microscopy confirmed the podocyte damage. Podocyte injury was accompanied by renal reduced nicotinamide-adenine dinucleotide phosphate oxidase activation, increased oxidative stress, and enhanced expression of aldosterone effector kinase Sgk1. Treatment with eplerenone, a selective aldosterone receptor blocker, almost completely prevented podocyte damage and proteinuria, with normalization of elevated reduced nicotinamide-adenine dinucleotide phosphate oxidase activity. In addition, proteinuria, podocyte damage, and Sgk1 upregulation were significantly alleviated by tempol, a membrane-permeable superoxide dismutase, suggesting the pathogenic role of oxidative stress. Although hydralazine treatment almost normalized blood pressure, it failed to improve proteinuria and podocyte damage. In cultured podocytes with consistent expression of mineralocorticoid receptor, aldosterone stimulated membrane translocation of reduced nicotinamide-adenine dinucleotide phosphate oxidase cytosolic components and oxidative stress generation in podocytes. Furthermore, aldosterone enhanced the expression of Sgk1, which was inhibited by mineralocorticoid receptor antagonist and tempol. In conclusion, podocytes are injured at the early stage in aldosterone-infused rats, resulting in the occurrence of proteinuria. Aldosterone can directly modulate podocyte function, possibly through the induction of oxidative stress and Sgk1.


Design, synthesis and biological evaluation of novel DCLK1 inhibitor containing purine skeleton for the treatment of pancreatic cancer.
Pancreatic cancer is a highly lethal form of malignancy that continues to pose a significant and unresolved health challenge. Doublecortin-like kinase 1 (DCLK1), a serine/threonine kinase, is found to be overexpressed in pancreatic cancer and holds promise as a potential therapeutic target for this disease. However, few potent inhibitors have been reported currently. Herein, a series of novel purine, pyrrolo [2,3-d]pyrimidine, and pyrazolo [3,4-d] pyrimidine derivatives were designed, synthesized, and evaluated their biological activities in vitro. Among them, compound I-5 stood out as the most potent compound with strong inhibitory activity against DCLK1 (IC50 = 171.3 nM) and remarkable antiproliferative effects on SW1990 cell lines (IC50 = 0.6 μM). Notably, I-5 exhibited higher in vivo antitumor potency (Tumor growth inhibition value (TGI): 68.6 %) than DCLK1-IN-1 (TGI: 24.82 %) in the SW1990 xenograft model. The preliminary mechanism study demonstrated that I-5 not only inhibited SW1990 cell invasion and migration, but also decreased the expression of prominin-1 (CD133) and cluster of differentiation 44 (CD44), which are considered as differentiation markers for SW1990 stem cells. All the results indicated that I-5, a novel DCLK1 inhibitor, shows promise for further investigation in the treatment of pancreatic cancer.


Parental Selection of Preschool Programming in Saudi Arabia: A Qualitative Study.
A global initiative has emerged in recent years to highlight early childhood education and care as a foundation for later learning and development. The goal of this study was to investigate the choices that Saudi Arabian parents made for their children's preschool education. In a qualitative, exploratory study, we investigated parental selection of preschool programming in Saudi Arabia. Guided by a phenomenological approach, we conducted semi-structured interviews with 12 Saudi Arabian women that had preschool-aged children. Although some research has examined the constraints and factors affecting parental selection of preschool programming, it remains unclear when these constraints and factors are considered by parents during the decision-making process, and how the decisions are made. The interviews revealed that parents consider not only educational reputation in the selection of preschool programming, but also instructional use of Arabic and English, religious foundation, teaching quality, classroom preparation, program scheduling, tuition costs, and physical security of the preschool environment. The results suggest that improving information collection is essential for empowering parents to make wise decisions, with the child's happiness and academic growth having a significant impact on those decisions. The discussion addresses the importance of identifying the factors that affect parental selection of preschool programming, so that educational professionals may better meet the needs and expectations of parents in Saudi Arabia.


Psychological Interventions for Post-traumatic Stress Symptoms in Psychosis: A Systematic Review of Outcomes.
Individuals with severe mental health problems, such as psychosis, are consistently shown to have experienced high levels of past traumatic events. They are also at an increased risk of further traumatisation through victimization events such as crime and assault. The experience of psychosis itself and psychiatric hospitalization have also been recognized to be sufficiently traumatic to lead to the development of post-traumatic stress (PTS) symptoms. Rates of post-traumatic stress disorder (PTSD) are elevated in people with psychosis compared to the general population. The current guidance for the treatment of PTSD is informed by an evidence base predominately limited to populations without co-morbid psychiatric disorders. The systematic review therefore sought to present the current available literature on the use of psychological treatments targeting PTS symptoms in a population with a primary diagnosis of a psychotic disorder. The review aimed to investigate the effect of these interventions on PTS symptoms and also the effect on secondary domains such as psychotic symptoms, affect and functioning. Fifteen studies were identified reporting on cognitive behavior therapy, prolonged exposure, eye movement desensitisation and reprocessing and written emotional disclosure. The review provides preliminary support for the safe use of trauma-focused psychological interventions in groups of people with severe mental health problems. Overall, the interventions were found to be effective in reducing PTS symptoms. Results were mixed with regard to secondary effects on additional domains. Further research including studies employing sufficiently powered methodologically rigorous designs is indicated.


[How do we explain the antihypertensive effect of bromocriptine? Clinical and experimental contribution].
The mechanism of the antihypertensive properties of bromocriptine was investigated in dog and man. In anaesthetized dogs, bromocriptine induced a decrease in the pressor responses to adrenaline or phenylephrine and reduced the hypotensive properties of clonidine. In 6 hypertensive patients with Parkinson's disease, chronic treatment with bromocriptine (46.4 +/- 12.0 mg/day) elicited both a significant decrease in blood pressure and an improvement of extrapyramidal symptoms. During bromocriptine therapy, domperidone (60 mg/day during 1 month), a specific peripheral dopaminergic antagonist was introduced. The drug did not change the levels of blood pressure (before domperidone: 150.8 +/- 22.5/79.9 +/- 8.9 mmHg after domperidone: 150 +/- 17.3/87.5 +/- 12.9 mmHg). These results confirm the antihypertensive properties of bromocriptine and demonstrate that, beside their dopaminergic properties, the drug also possesses mixed alpha 1 and alpha 2 adrenolytic actions. These two properties may contribute to the mechanism of the antihypertensive effects of bromocriptine. These data also show that domperidone can be used in bromocriptine-treated hypertensive parkinsonians without side effects on blood pressure.


[A study on measurement of hardness of oral mucosal lesions].
This study was designed to estimate objectively the hardness of the oral mucosa which is not directly supported by the bone. A new, easy-to-use hardness-measurement apparatus for measuring the oral mucosa free of the underlining bone was manufactured for trial use. The hardness values of the oral mucosa were evaluated quantitatively, resulting from pressing 2 mm into the mucosa with a 2-mm-in-diameter hand-held probe coupled to the hardness-measurement apparatus. Experimental and clinical confirmation was made that practical errors in hardness values are quite permissible when using this apparatus. This fool-proof method was further examined in extensive clinical application. The normal mucous membrane of the tongue, the floor of the mouth, and the cheek respectively measure 4.45, 4.33, and 2.96 gf on average with this apparatus. Examining 26 cases of squamous cell carcinoma on the tongue, the floor of the mouth or the buccal mucosa (16.87-80.32 gf) revealed that the value of measurement of squamous cell carcinoma was as much as 2.31-19.06 times in comparison with the normal mucous membrane. Squamous cell carcinomas were ascertained to be statistically harder than the normal mucosa (p < 0.001) and the tumor margins may be far more precisely clarified with this apparatus than with the human finger alone.


Risk adjustment for health status.
A managed care plan's capitation payments should include an adjustment for its members' health status. Demographic and diagnostic information can be used to make the adjustment, which would help to minimize a health plan's potential financial liabilities. Without the adjustment, health plans have an incentive not to enroll persons with the heaviest care needs.


Effect of cultural system and essential oil treatment on antioxidant capacity in raspberries.
The effects of cultural system and essential oil treatment on antioxidant capacities in raspberries were evaluated. Raspberries were hand-harvested from organic and conventional farms in Maryland, USA, and were treated with essential oil including carvacrol, anethole, cinnamic acid, perillaldehyde, cinnamaldehyde, and linalool. Results from this study showed that raspberries grown from organic culture exhibited higher value of antioxidant capacities and individual flavonoids contents. Moreover, the organic culture also enhanced the activities of antioxidant enzymes. In addition, essential oil treatments promoted the antioxidant enzymes activities and antioxidant capacities of raspberries, and the most effective compound was perillaldehyde. In conclusion, raspberries produced from organic culture contained significantly higher antioxidant capacities than those produce from conventional culture. Postharvest essential oil treatments have positive effect on enhancing antioxidant capacities in raspberries from both organic and conventional cultures.


Embodied Information in Cognitive Tasks: Haptic Weight Sensations Affect Task Performance and Processing Style.
Research in the field of embodied cognition showed that incidental weight sensations influence peoples' judgments about a variety of issues and objects. Most studies found that heaviness compared to lightness increases the perception of importance, seriousness, and potency. In two experiments, we broadened this scope by investigating the impact of weight sensations on cognitive performance. In Experiment 1, we found that the performance in an anagram task was reduced when participants held a heavy versus a light clipboard in their hands. Reduced performance was accompanied by an increase in the perceived effort. In Experiment 2, a heavy clipboard elicited a specific response heuristic in a two-alternative forced-choice task. Participants showed a significant right side bias when holding a heavy clipboard in their hands. After the task, participants in the heavy clipboard condition reported to be more frustrated than participants in the light clipboard condition. In both experiments, we did not find evidence for mediated effects that had been proposed by previous literature. Overall, the results indicate that weight effects go beyond judgment formation and highlight new avenues for future research.


Reliability and accuracy in three-dimensional gait analysis: a comparison of two lower body protocols.
The standard Plug-in-Gait (PiG) protocol used in three-dimensional gait analysis is prone to errors arising from inconsistent anatomical landmark identification and knee axis malalignment. The purpose of this study was to estimate the reliability and accuracy of a custom made lower body protocol (MA) compared with the PiG protocol. Twenty-five subjects volunteered to evaluate the intertrial reliability. In addition, intersession reliability was examined in 10 participants. An indirect indicator of accuracy according to the knee varus/valgus and flexion/extension range of motion (ROM) was used. Regarding frontal plane knee angles and moments as well as transverse plane motions in the knee and hip joint, the intersession errors were lower for the MA compared with the standard approach. In reference to the knee joint angle cross-talk, the MA produced 4.7° more knee flexion/extension ROM and resulted in 6.5° less knee varus/valgus ROM in the frontal plane. Therefore, the MA tested in this study produced a more accurate and reliable knee joint axis compared with the PiG protocol. These results are especially important for measuring frontal and transverse plane gait parameters.


Prediction of 15 N chemical shifts by machine learning.
We demonstrate the potential for machine learning systems to predict three-dimensional (3D)-relevant NMR properties beyond traditional 1 H- and 13 C-based data, with comparable accuracy to density functional theory (DFT) (but orders of magnitude faster). Predictions of DFT-calculated 15 N chemical shifts for 3D molecular structures can be achieved using a machine learning system-IMPRESSION (Intelligent Machine PREdiction of Shift and Scalar information Of Nuclei), with an accuracy of 6.12-ppm mean absolute error (∼1% of the δ15 N chemical shift range) and an error of less than 20 ppm for 95% of the chemical shifts. It provides less accurate raw predictions of experimental chemical shifts, due to the limited size and chemical space diversity of the training dataset used in its creation, coupled with the limitations of the underlying DFT methodology in reproducing experiment.


Lesion kinetics in a non-human primate model of endometriosis.
Endometriosis is a common cause of pelvic pain and infertility in women of reproductive age. It is characterized by the presence of endometrial tissue outside the normal location, predominantly in the pelvic peritoneum causing severe abdominal pain. However, the severity of the symptoms of endometriosis does not always correlate with the anatomic severity of the disease. This lack of correlation may be due to morphological lesion variation during disease progression. This study examined lesion kinetics in a non-human primate model of endometriosis to better understand lesion dynamics. Endometriosis was experimentally induced in nine normal cycling female adult olive baboons (Papio anubis) by i.p. inoculation of autologous menstrual endometrium on Day 2 of menses for two consecutive menstrual cycles. Diagnostic laparoscopies were performed between Day 8-12 post-ovulation at 1, 3, 6, 9 and 12 months, followed by a necropsy at 15 months, after the second inoculation. In two animals, lesions were excised/ablated at 6 months and they were monitored for lesion recurrence and morphological changes by serial laparoscopy. Furthermore, five control animals underwent surgeries conducted at the same time points but without inoculation. A total of 542 endometriotic lesions were observed. The location, macroscopic (different colours) and microscopic appearance confirmed distinct endometriosis pathology in line with human disease. The majority of the lesions found 1 month after tissue inoculation were red lesions, which frequently changed colour during the disease progression. In contrast, blue lesions remained consistently blue while white lesions were evident at the later stages of the disease process and often regressed. There were significantly lower numbers of powder burn, blister and multicoloured lesions observed per animal in comparison to black and blue lesions (P-value≤0.05). New lesions were continually arising and persisted up to 15 months post-inoculation. Lesions reoccurred as early as 3 months after removal and 69% of lesions excised/ablated had reoccurred 9 months later. Interestingly, endometriotic lesions were also found in the non-inoculated animals, starting at the 6-month time point following multiple surgeries. Documentation of lesion turnover in baboons indicated that lesions changed their colour from red to white over time. Different lesion types underwent metamorphosis at different rates. A classification of lesions based on morphological appearance may help disease prognosis and examination of the effect of the lesion on disease symptoms, and provide new opportunities for targeted therapies in order to prevent or treat endometriosis. Surgical removal of endometriotic lesions resulted in a high incidence of recurrence. Spontaneous endometriosis developed in control baboons in the absence of inoculation suggesting that repetitive surgical procedures alone can induce the spontaneous evolution of the chronic disease. Although lesion excision/ablation may have short-term benefits (e.g. prior to an IVF cycle in subfertile women), for long-term relief of symptoms perhaps medical therapy is more effective than surgical therapy.


Trigeminal neuropathy in dogs: a retrospective study of 29 cases (1991-2000).
The medical records of 29 dogs unable to close their mouths due to flaccid paralysis or paresis of the muscles innervated by the mandibular branch of the trigeminal nerve, were reviewed. Idiopathic trigeminal neuropathy was diagnosed in 26 dogs based on complete resolution of clinical signs and lack of any long-term neurological disease. Of these dogs, golden retrievers were overrepresented. No age, sex, or seasonal predispositions were identified. Trigeminal sensory innervation deficits were observed in 35% (9/26), facial nerve deficits were observed in 8% (2/26), and Horner's syndrome was observed in 8% (2/26) of dogs. Electromyographic examination of the muscles of mastication revealed abnormalities in seven of nine dogs. Results of cerebrospinal fluid analysis were abnormal in seven of eight dogs. Corticosteroid therapy did not affect the clinical course of the disease. Mean time to recovery was 22 days. Lymphosarcoma, Neospora caninum infection, and severe polyneuritis of unknown origin were diagnosed in three of 29 dogs at necropsy.


Current and potential utility of transcranial magnetic stimulation in the diagnostics before brain tumor surgery.
This article describes the evolution and state-of-the-art of navigated transcranial magnetic stimulation for evaluation of patients with brain tumors in presumed eloquent location. Alternative noninvasive technologies for functional brain mapping are described and assessed in the context of their usability and clinical needs. In addition to the description of the current validation level and clinical application of navigated transcranial magnetic stimulation for motor and language mapping, the manuscript highlights ongoing research efforts and provides an outlook on upcoming developments in the field of noninvasive brain mapping. Finally, the clinical rationale for presurgical noninvasive brain mapping is discussed in the light of current developments in neurosurgery.


Inactive Hepatitis B Carrier and Pregnancy Outcomes: A Systematic Review and Meta-analysis.
We aimed to explore whether maternal asymptomatic hepatitis B (HB) infection effects on pre-term rupture of membranous (PROM), stillbirth, preeclampsia, eclampsia, gestational hypertension, or antepartum hemorrhage. We searched the PubMed, Scopus, and ISI web of science from 1990 to Feb 2015. In addition, electronic literature searches supplemented by searching the gray literature (e.g., conference abstracts thesis and the result of technical reports) and scanning the reference lists of included studies and relevant systematic reviews. We explored statistical heterogeneity using the, I2 and tau-squared (Tau2) statistical tests. Eighteen studies were included. Preterm rupture of membranous (PROM), stillbirth, preeclampsia, eclampsia, gestational hypertension and antepartum hemorrhage were considerable outcomes in this survey. The results showed no significant association between inactive HB and these complications in pregnancy. The small amounts of P-value and chi-square and large amount of I2 suggested the probable heterogeneity in this part, which we tried to modify with statistical methods such as subgroup analysis. Inactive HB infection did not increase the risk of adversely mentioned outcomes in this study. Further, well-designed studies should be performed to confirm the results.


The association between patient-reported incidents in hospitals and estimated rates of patient harm.
The aim of this study was to test the association between the rates of patient-reported incidents and patient harm documented in the patient record. The study was a secondary analysis of two national hospital assessments conducted in 2011. Hospital services in Norway. The patient survey was a standard national patient-experience survey conducted at the hospital level for all 63 hospitals in Norway. The medical record review was performed by 47 Global Trigger Tools (GTTs) in all 19 hospital trusts and 4 private hospitals. The two data sets were matched at the unit level, yielding comparable patient experiences and GTT data for 7 departments, 16 hospitals and 11 hospital trusts. No intervention. The correlation at the unit level between the patient-reported incident in hospital instrument (PRIH-I) and estimated rates of patient harm from the GTT. The PRIH-I index was significantly correlated with all patient-reported experience indicators at the individual level, with estimates for all patient harm events (Categories E-I) at the unit level (r = 0.62, P < 0.01), and with estimates of more serious harm events in Categories F-I (r = 0.42, P < 0.05). Patient-reported incidents in hospitals, as measured by the PRIH-I, are strongly correlated with patient harm rates based on the GTT. This indicates that patient-reported incidents are related to patient safety, but more research is needed to confirm the usefulness of patient reporting in the evaluation of patient safety.


In vivo study of NMDA-sensitive glutamate receptor by fluorothienylcyclohexylpiperidine [correction of fluorothienylcycloexylpiperidine], a possible ligand for positron emission tomography.
As a preliminary to positron emission tomography (PET) studies of excitatory amino acid neurotransmission, N-methyl-D-aspartate (NMDA)-sensitive glutamate receptors of mice and rats were labelled in vivo with [3H]fluorothienylcyclohexylpiperidine [corrected] (FTCP), which binds to the phencyclidine site of the NMDA receptor. After intravenous injection, the half-life of clearance of authentic FTCP from blood was 4.2 min in mice, 12 min in rats and 45 min in a rhesus monkey. In rodent brain, the specific binding of [3H]FTCP, 10 min after intravenous injection, was 10-20% of the total binding and no regional differences were observed. However, if animals were treated with NMDA intraperitoneally (0.68 mmol/kg), 10 min before injection of [3H]FTCP, a three- to five-fold increase in specific binding was observed in hippocampus, cerebral cortex and striatum but not in cerebellum. Thus, specific binding of [3H]FTCP in vivo revealed the physiological status of the NMDA receptor; in fact, preliminary PET studies with [18F]FTCP in monkeys indicated increased binding after activation of NMDA receptors. These data suggest that PET with [18F]FTCP can be a tool to evaluate physiological or pathological modifications of the function of NMDA receptors.


The relationship between dispositional empathy and prefrontal cortical functioning in patients with frontal lobe epilepsy.
Rehabilitation focuses brain-behavior relationship which highlights interaction between psychological and neurobiological factors for better patient care. There is a missing link in the literature about socio-cognitive aspects of frontal lobe epilepsy. Our objective was to examine prefrontal cortical functioning (PCF) and empathic abilities in patients with frontal lobe epilepsy (FLE). Further, we analyzed whether any relationship between components of dispositional empathy and PCF exists in patients with FLE. The study was designed in an experimental paradigm. Sixty patients with FLE were recruited from Sheikh Zayed and Jinnah hospital, Pakistan. Sixty healthy individuals in response to an advertisement took part in the study as control subjects. Participants completed interpersonal reactivity index. Following they performed clock drawing test and word-color identification task switching experiment. Patients with FLE demonstrated weaker PCF (i.e., cognitive flexibility and executive function) as compared to healthy control subjects. Patients with FLE scored lesser on cognitive empathy as compared to healthy control subjects. On contrary, there was no significant difference between patient and control group on affective empathy. Cognitive not affective empathy was potential predictor of PCF. Cognitive empathy is a significant marker of prefrontal cortical functioning (PCF) in FLE. Higher cognitive empathy would lead to efficient PCF.


Involvement of a common 10-amino-acid segment in the cytoplasmic region of CD40 but different MAP kinases in different CD40-mediated responses.
CD40-mediated signals can induce cell aggregation, proliferation and rescue from apoptosis in WEHI231. To define which segment of cytoplasmic domain of CD40 and how signals are involved in those events, we generated mutant CD40 transfectants. We demonstrated the same 10 amino acid segment that could bind to tumor necrosis factor receptor associated factor-2 and -3 mediated all those responses. However, activation pattern of mitogen activated protein kinases was different. Immunoglobulin M-mediated apoptosis was inhibited by CD40-mediated signal that activated c-Jun aminoterminal kinase synergistically. While, CD40 stimulus through the 10 amino acid segment alone that induced cell aggregation and proliferation resulted in activation of extracellular signal-regulated protein kinase 2.


Biorhizome: A Biosynthetic Platform for Colchicine Biomanufacturing.
Colchicine is one of the oldest plant-based medicines used to treat gout and one of the most important alkaloid-based antimitotic drugs with anticancer potential, which is commercially extracted from Gloriosa superba. Clinical trials suggest that colchicine medication could prevent atrial fibrillation recurrence after cardiac surgery. In addition, therapeutic colchicine is undergoing clinical trials to treat non-diabetic metabolic syndrome and diabetic nephropathy. However, the industrial-scale biomanufacturing of colchicine have not yet been established. Clearly, further studies on detailed biorhizome-specific transcriptome analysis, gene expression, and candidate gene validation are required before uncover the mechanism of colchicine biosynthesis and biorhizome-based colchicine biomanufacturing. Annotation of 32312 assembled multiple-tissues transcripts of G. superba represented 15088 unigenes in known plant specific gene ontology. This could help understanding colchicine biosynthesis in G. superba. This review highlights the biorhizomes, rhizome specific genes or gene what expressed with high level in rhizomes, and deep fluid dynamics in a bioreactor specifically for the biomanufacture of colchicine.


Bulimic vomiting alters pain tolerance and mood.
Bulimia, a disorder of episodic binging and purging, remains without a known etiology. A case report is presented of a patient who attributed bulimic episodes to efforts at inducing euphoria. Experimental pain tolerance was increased by bulimic vomiting, blocked by naloxone, but not by saline. Vomiting was also associated with falls in depression and anxiety. Plasma ACTH and cortisol, putative markers for beta-endorphin, also rose following vomiting. It is hypothesized that in some bulimics, the disorder arises by virtue of an addiction to one's own internally released endogenous opioid peptides.


Office-Based Subglottic Evaluation in Children With Risk of Subglottic Hemangioma.
Children with V3 cutaneous infantile hemangiomas (IH) and PHACE syndrome have a high incidence for airway hemangioma, 29% and 52%, respectively. Therefore, a clinical evaluation for these high-risk children is essential. We report our experience with in-office lower airway evaluation (OLAE) in these high-risk children. Since 2003, 5 children with IH of the V3 cutaneous distribution and 3 children with PHACE syndrome underwent OLAE. Average age of presentation was 2.75 months. Two children had stridor at initial evaluation, and 1 child had subglottic hemangioma. This child was evaluated serially with OLAE to monitor disease progression and treatment response. A total of 10 upper tracheoscopies were performed on the 8 patients without respiratory complications. An airway evaluation is essential to evaluate and manage this high-risk population. Typically, operative endoscopy requires general anesthesia. However, in these high-risk children, we have performed OLAE without sedation to evaluate the trachea. High-speed recording and playback is essential in this method. Our series demonstrates that awake OLAE is possible and may be a safe technique to evaluate and monitor disease progression in these high-risk patients. These patients avoided general anesthesia and delay in diagnosis and did not incur any complications during or after OLAE.


Lower limb ischemia.
Diabetes mellitus is the major cause of nontraumatic amputations in the industrialized countries. In 1980 the scientific societies of diabetology and the patients' associations subscribed at St. Vincent a declaration together with the WHO, where a 50% decrease in amputations was among the primary objectives of clinical diabetology. The main cause of amputations is foot ischemia related to obliterating peripheral arterial disease, more diffuse and severe in diabetics, typically affecting the infrapopliteal vessels. Clinical patterns of foot ischemia include superficial necrotic areas and gangrene involving the entire foot with necrosis of all anatomic planes. Infection is a common complication with a negative impact on prognosis. Definitive treatment of ischemia is achieved with revascularization which at present due to the advances in diagnostic procedures can be performed in diabetics with the same effectiveness observed in non diabetics.


Error detection across the adult lifespan: Electrophysiological evidence for age-related deficits.
With increasing age, cognitive control processes steadily decline. Prior research suggests that healthy older adults have a generally intact performance monitoring system, but show specific deficits in error awareness, i.e., the ability to detect committed errors. We examined the neural processing of errors across the adult lifespan (69 participants; age range 20-72 years) by analysing the error (-related) negativity (Ne/ERN) and the error positivity (Pe) using an adapted version of the Go/Nogo task. At a stable overall error rate, higher age was associated with a greater proportion of undetected errors. While the Ne/ERN was associated with the processing of errors in general, the Pe amplitude was modulated by detected errors only. Furthermore, the Pe amplitude for detected errors was significantly smaller in older adults, in contrast to the Ne/ERN amplitude which did not show age-related changes. Structural path models suggested that through those age-related changes in Pe amplitude, an indirect effect on the performance was observed. Our results confirm and extend previous extreme-group based findings about specific deficits in error detection associated with higher age using age as a continuous predictor. Age-related reductions in Pe amplitude, associated with more undetected errors, are independent of early error processing, as evidenced by the preserved Ne/ERN.


PDV1 and PDV2 mediate recruitment of the dynamin-related protein ARC5 to the plastid division site.
During plastid division, the dynamin-related protein ACCUMULATION AND REPLICATION OF CHLOROPLASTS5 (ARC5) is recruited from the cytosol to the surface of the outer chloroplast envelope membrane. In Arabidopsis thaliana arc5 mutants, chloroplasts arrest during division site constriction. Analysis of mutants similar to arc5 along with map-based cloning identified PLASTID DIVISION1 (PDV1), an integral outer envelope membrane protein, and its homolog PDV2 as components of the plastid division machinery. Similar to ARC5, PDV1 localized to a discontinuous ring at the division site in wild-type plants. The midplastid PDV1 ring formed in arc5 mutants and the ARC5 ring formed in pdv1 and pdv2 mutants, but not in pdv1 pdv2. Stromal FtsZ ring assembly occurred in pdv1, pdv2, and pdv1 pdv2, as it does in arc5. Topological analysis showed that the large N-terminal region of PDV1 upstream of the transmembrane helix bearing a putative coiled-coil domain is exposed to the cytosol. Mutation of the conserved PDV1 C-terminal Gly residue did not block PDV1 insertion into the outer envelope membrane but did abolish its localization to the division site. Our results indicate that plastid division involves the stepwise localization of FtsZ, PDV1, and ARC5 at the division site and that PDV1 and PDV2 together mediate the recruitment of ARC5 to the midplastid constriction at a late stage of division.


The activation of human blood coagulation factor X on the surface of endothelial cells: a comparison with various vascular cells, platelets and monocytes.
Rates of factor X activation on endothelial cells were compared with activation rates on other vascular cells, platelets, monocytes and negatively charged phospholipid vesicles. Factor VIIa-mediated factor X activation was observed on smooth muscle cells and fibroblasts in the absence of cell-perturbing agents, whereas endothelial cells required activation in order to allow extrinsic activation of factor X. On the other hand, unperturbed endothelial cells did promote intrinsic, factor VIII/IXa-dependent activation of factor X. The rate of factor X activation on these cells was about one-sixth of that on ionophore A23187-stimulated platelets. Also, smooth muscle cells and fibroblasts were able to activate factor X through the intrinsic pathway, although to a lesser extent than endothelial cells. Monocytes were ineffective in this respect. Prothrombin fragment 1, the prothrombin fragment containing the gamma-carboxyglutamic acid domain known to mediate binding of vitamin K-dependent coagulation factors to phospholipid surfaces, inhibited factor VIII/IXa-dependent factor X activation on endothelial cells (IC50 3.2 microM) to a lesser extent than on phospholipid vesicles (IC50 0.2 microM). Therefore, besides negatively charged phospholipids, other membrane constituents seem to be involved in endothelial cell mediated, intrinsic activation of factor X. Perturbation of endothelial cells with phorbol myristate acetate (PMA) or lipopolysaccharide (LPS) was without effect on intrinsic activation of factor X. This observation indicates that membrane constituents of endothelial cells involved in factor VIII/IXa-dependent activation of factor X are constitutively expressed.


Stomatal clustering in Begonia associates with the kinetics of leaf gaseous exchange and influences water use efficiency.
Stomata are microscopic pores formed by specialized cells in the leaf epidermis and permit gaseous exchange between the interior of the leaf and the atmosphere. Stomata in most plants are separated by at least one epidermal pavement cell and, individually, overlay a single substomatal cavity within the leaf. This spacing is thought to enhance stomatal function. Yet, there are several genera naturally exhibiting stomata in clusters and therefore deviating from the one-cell spacing rule with multiple stomata overlaying a single substomatal cavity. We made use of two Begonia species to investigate whether clustering of stomata alters guard cell dynamics and gas exchange under different light and dark treatments. Begonia plebeja, which forms stomatal clusters, exhibited enhanced kinetics of stomatal conductance and CO2 assimilation upon light stimuli that in turn were translated into greater water use efficiency. Our findings emphasize the importance of spacing in stomatal clusters for gaseous exchange and plant performance under environmentally limited conditions.


Nickel-Catalyzed Direct C (sp(3))-H Arylation of Aliphatic Amides with Thiophenes.
Nickel-catalyzed heteroarylation of the inactive methyl C(sp(3))-H bond of aliphatic amide with heteroarenes is described. The method takes advantage of chelation assistance by an 8-aminoquinolinyl moiety. The synthetic reaction has good tolerance toward functional groups, and it can be used in the construction of various kinds of alkyl-substituted heteroarenes.


Cutometrical measurement confirms the efficacy of the composite skin grafting using allogeneic acellular dermis in burns.
Measurement of viscoelasticity of the skin using the device called cutometer seems to be a very precise and promising diagnostic method in different skin pathologies. However, only a very limited number of reports dealing with the use of cutometer in burns is available. The composite skin grafting using an allogeneic acellular dermis covered with a thin epidermal autologous graft is supposed to improve viscoelastic properties of grafted skin in full-thickness burns. Cutometer MPA 580 has been used for examination of healthy skin, healed and maturated autologous split-thickness skin grafts, and in two patiens 6 years after composite skin grafting using the allogeneic acellular dermis + thin autologous epidermal graft in one stage. Typical curves defined by values of R0-R9 parameters are presented. These data document significantly better viscoelastic properties of composite skin grafts than those of conventional split-thickness skin grafts after 6 years. Measurements of viscoelasticity of the skin, using a cutometer, will probably bring useful and precise data for evaluation of the reconstructed skin in burns. Significantly better properties of the composite skin graft were assessed clinically and measured by Cutometer MPA 580 6 years after surgery.


Rapid magnetic cell delivery for large tubular bioengineered constructs.
Delivery of cells into tubular tissue constructs with large diameters poses significant spatial and temporal challenges. This study describes preliminary findings for a novel process for rapid and uniform seeding of cells onto the luminal surface of large tubular constructs. Fibroblasts, tagged with superparamagnetic iron oxide nanoparticles (SPION), were directed onto the luminal surface of tubular constructs by a magnetic field generated by a k4-type Halbach cylinder device. The spatial distribution of attached cells, as measured by the mean number of cells, was compared with a conventional, dynamic, rotational cell-delivery technique. Cell loading onto the constructs was measured by microscopy and magnetic resonance imaging. The different seeding techniques employed had a significant effect on the spatial distribution of the cells (p < 0.0001). The number of attached cells at defined positions within the same construct was significantly different for the dynamic rotation technique (p < 0.05). In contrast, no significant differences in the number of cells attached to the luminal surface were found between the defined positions on the construct loaded with the Halbach cylinder. The technique described overcomes limitations associated with existing cell-delivery techniques and is amenable to a variety of tubular organs where rapid loading and uniform distribution of cells for therapeutic applications are required.


Prostaglandin E2 inhibits BMP signaling and delays chondrocyte maturation.
While cyclooxygenases are important in endochondral bone formation during fracture healing, mechanisms involved in prostaglandin E2 (PGE2) regulation of chondrocyte maturation are incompletely understood. The present study was undertaken to determine if PGE2 effects on chondrocyte differentiation are related to modulation of the bone morphogenetic protein (BMP) signaling pathway. In primary murine sternal chondrocytes, PGE2 differentially regulated genes involved in differentiation. PGE2 induced type II collagen and MMP-13, had minimal effects on alkaline phosphatase, and inhibited the expression of the maturational marker, type X collagen. In BMP-2-treated cultures, PGE2 blocked the induction of type X collagen. All four EP receptors were expressed in chondrocytes and tended to be inhibited by BMP-2 treatment. RCJ3.1C5.18 chondrocytes transfected with the protein kinase A (PKA) responsive reporter, CRE-luciferase, showed luciferase induction following exposure to PGE2, consistent with activation of PKA signaling and the presence of the EP2 and EP4 receptors. Both PGE2 and the PKA agonist, dibutyryl cAMP, blocked the induction of the BMP-responsive reporter, 12XSBE, by BMP-2 in RCJ3.1C5.18 chondrocytes. In contrast, PGE2 increased the ability of TGF-beta to activate the TGF-beta-responsive reporter, 4XSBE. Finally, PGE2 down-regulated BMP-mediated phosphorylation of Smads 1, 5, and 8 in RCJ3.1C5.18 cells and in primary murine sternal chondrocytes. Altogether, the findings show that PGE2 regulates chondrocyte maturation in part by targeting BMP/Smad signaling and suggest an important role for PGE2 in endochondral bone formation.