Time-resolved three-dimensional imaging of the left atrium and pulmonary veins in the interventional suite--a comparison between multisweep gated rotational three-dimensional reconstructed fluoroscopy and multislice computed tomography.
Cardiac computed tomography (CT) is commonly used to visualize left atrial (LA) anatomy for ablation of atrial fibrillation. We have developed a new imaging technique that allows acquisition and visualization of three-dimensional (3D) cardiac images in the catheter lab. We sought to compare LA and pulmonary vein (PV) dimensions acquired using gated multisweep rotational fluoroscopy (C-arm CT) system and multislice computed tomography (MSCT) in an in vivo porcine model. A Siemens AXIOM Artis dTA C-arm system (Siemens AG, Medical Solutions) was modified to allow acquisition of four bidirectional sweeps during synchronized acquisition of the electrocardiogram signal to allow retrospective gating. C-arm CT image volumes were then reconstructed. Gated MSCT (SOMATOM Sensation 16 and 64, Siemens AG, Medical Solutions) and C-arm CT images were acquired in six animals. The two main PV diameters were measured in orthogonal axes. LA volumes were calculated. C-arm CT measurements were compared with the MSCT measurements. The average PV diameters using the C-arm CT were 2.24 x 1.35 cm, versus 2.27 x 1.38 cm for CT. The average difference was 0.034 cm (1.9%) between the C-arm CT and standard CT. The average LA volume using MSCT was 49.1 +/- 12.7 cm(3), as compared with 51.0 +/- 8.7 cm(3) obtained by the C-arm CT. The average difference between the C-arm CT and the MSCT was 1.9 cm(3) (3.7%). There were no significant differences in either the PV or LA measurements. Visualization of 3D cardiac anatomy during ablation procedures is possible and highly accurate. The 3D cardiac reconstructions acquired during ablation procedures will be valuable for procedural planning and guidance.


Descriptive Analysis of the Interactive Patterning of the Vocalization Subsystems in Healthy Participants: A Dynamic Systems Perspective.
Purpose Normative data for many objective voice measures are routinely used in clinical voice assessment; however, normative data reflect vocal output, but not vocalization process. The underlying physiologic processes of healthy phonation have been shown to be nonlinear and thus are likely different across individuals. Dynamic systems theory postulates that performance behaviors emerge from the nonlinear interplay of multiple physiologic components and that certain patterns are preferred and loosely governed by the interactions of physiology, task, and environment. The purpose of this study was to descriptively characterize the interactive nature of the vocalization subsystem triad in subjects with healthy voices and to determine if differing subgroups could be delineated to better understand how healthy voicing is physiologically generated. Method Respiratory kinematic, aerodynamic, and acoustic formant data were obtained from 29 individuals with healthy voices (21 female and eight male). Multivariate analyses were used to descriptively characterize the interactions among the subsystems that contributed to healthy voicing. Results Group data revealed representative measures of the 3 subsystems to be generally within the boundaries of established normative data. Despite this, 3 distinct clusters were delineated that represented 3 subgroups of individuals with differing subsystem patterning. Seven of the 9 measured variables in this study were found to be significantly different across at least 1 of the 3 subgroups indicating differing physiologic processes across individuals. Conclusion Vocal output in healthy individuals appears to be generated by distinct and preferred physiologic processes that were represented by 3 subgroups indicating that the process of vocalization is different among individuals, but not entirely idiosyncratic. Possibilities for these differences are explored using the framework of dynamic systems theory and the dynamics of emergent behaviors. A revised physiologic model of phonation that accounts for differences within and among the vocalization subsystems is described. Supplemental Material https://doi.org/10.23641/asha.7616462.


Effect of Increased Lumbar Lordosis on Lumbar Multifidus and Longissimus Thoracis Activation During Quadruped Exercise in Patients With Chronic Low Back Pain: An EMG Study.
The objective was to investigate the electromyographic activity of the lumbar multifidus (MF) muscle and longissimus thoracis muscle, along with their activity ratio (MF longissimus thoracis ratio), during quadruped stabilization exercise performed with neutral posture and with increased lumbar lordosis in patients with chronic low back pain (CLBP). A total of 23 patients with CLBP (12 females and 11 males) were recruited based on inclusion and exclusion criterion. Each patient performed 4 exercises in random order, with surface electromyography electrodes and an electrogoniometer attached. A cross-sectional study design was used to measure the amplitude of muscle activation (as a percentage of maximum voluntary contraction) in each patient across the 2 muscles (MF and longissimus thoracis) during quadruped stabilization exercise with neutral posture and with increased lumbar lordosis. A 2-way analysis of variance was conducted, which demonstrated a statistically significant increase in the recruitment of MF with increased lumbar lordosis in patients with CLBP during quadruped exercise. An increase of 9.7% and 16.9% maximum voluntary contraction in MF electromyographic activity was observed in lumbar lordosis posture during the quadruped leg raise and quadruped leg-arm raise exercise, respectively (P < .01), when compared to the neutral posture. The increased recruitment of MF with lumbar lordosis in the quadruped position has strong implications in the assessment and management of patients with CLBP.


Gender, creativity, depression, and attributional style in adolescents with high academic ability.
The present study examined the relationship among gender, creativity, depression, and attributional style among high-achieving adolescents. One hundred twenty-eight eighth- and ninth-grade high-achieving students completed the Torrance Test of Creative Thinking (TTCT), the Children's Depression Inventory (CDI), and the Children's Attribution Style Questionnaire--Revised (KASTAN-R CASQ). The results indicate that there were gender differences only on the verbal component of the TTCT, with females scoring significantly higher. For both sexes, there was a significant relationship between figural creativity and a depressogenic attributional style. However, for females, high verbal creativity was associated with low levels of depression and a positive attributional style.


Influence of Acetabular Labral Tear Length on Outcomes After Hip Arthroscopy for Femoroacetabular Impingement Syndrome With Capsular Plication.
The literature on the effects of labral tear on patient-reported outcomes, midterm pain, and overall patient satisfaction is limited. To determine the effect of labral tear length on postoperative outcomes after hip arthroscopy for femoroacetabular impingement syndrome (FAIS). Cohort study; Level of evidence, 3. Consecutive patients undergoing primary hip arthroscopy for FAIS from January 2012 to January 2016 were identified in a prospectively collected database. All patients completed the Hip Outcome Score-Activities of Daily Living (HOS-ADL), Hip Outcome Score-Sports Subscale (HOS-SS), modified Harris Hip Score (mHHS), and visual analog scale for pain and satisfaction. Patients were stratified by labral tear length into small (<2.5 cm) or large (≥2.5 cm) based on the receiver operating characteristic curve analysis. Patient characteristics and outcomes were analyzed with multivariate linear regression analyses to identify predictors of labral tear length. Binary logistic regression analysis was performed to determine whether labral tear length predicted the likelihood of achieving the minimal clinically important difference. Of the 747 eligible patients, 600 (80.3%) had 2-year reported outcomes and were included in the final analysis. Mean age, body mass index, and tear length were 33.5 ± 12.3 years, 25.4 ± 9.2 kg/m2, and 2.7 ± 0.7 cm, respectively. Men had higher frequency of large tears when compared with women (77% vs 43.7%, P < .001). Independent t test demonstrated significant differences in 2-year outcomes between patients with tears <2.5 and ≥2.5 cm, respectively: HOS-ADL (87.3 ± 16.3 vs 84.3 ± 18.1, P = .033), HOS-SS (76.6 ± 23.5 vs 70.5 ± 27.7, P = .005), mHHS (82.5 ± 18.0 vs 78.5 ± 18.2, P = .009), and satisfaction (83.5 ± 23.4 vs 77.8 ± 34.9, P = .026). Binary logistic regression analysis demonstrated that labral tear length is an independent predictor of visual analog scale for satisfaction, HOS-ADL, HOS-SS, and mHHS. Binary logistic regression analysis demonstrated that patients with small labral tears had a higher likelihood of achieving the minimal clinically important difference for the HOS-SS (odds ratio, 1.61; 95% CI, 1.39-1.92; P < .02) and the patient acceptable symptomatic state for the mHHS (odds ratio, 1.56; 95% CI, 1.11-2.2; P = .038) than those with larger tears. Labral tear length is independently predictive of patient-reported outcomes after hip arthroscopy for FAIS. Furthermore, patients with smaller tears (<2.5 cm) had better outcomes and a higher likelihood of achieving a minimal clinically important difference at 2-year follow-up. However, the mean differences between changes in pre- and postoperative outcomes were relatively small and may not be clinically meaningful.


Strategic Use of Affiliative Vocalizations by Wild Female Baboons.
Although vocal production in non-human primates is highly constrained, individuals appear to have some control over whether to call or remain silent. We investigated how contextual factors affect the production of grunts given by wild female chacma baboons, Papio ursinus, during social interactions. Females grunted as they approached other adult females 28% of the time. Supporting previous research, females were much more likely to grunt to mothers with young infants than to females without infants. Grunts also significantly increased the likelihood of affiliative interactions among all partners. Notably, however, grunts did not simply mirror existing social bonds. Instead, they appeared to perform a very different function: namely, to serve as signals of benign intent between partners whose relationship is not necessarily close or predictable. Females were less likely to grunt to their mothers or adult daughters-the individuals with whom they shared the closest and least aggressive bonds-than to other females. In contrast, patterns of grunting between sisters were similar to those between nonkin, perhaps reflecting sisters' more ambivalent relationships. Females grunted at higher rates to lower-ranking, than to higher-ranking, females, supporting the hypothesis that grunts do not simply signal the signaler's level of arousal or anxiety about receiving aggression, but instead function as signals of benign intent. Taken together, results suggest that the grunts given by female baboons serve to reduce uncertainty about the likely outcome of an interaction between partners whose relationship is not predictably affiliative. Despite their limited vocal repertoire, baboons appear to be skilled at modifying call production in different social contexts and for different audiences.


Genetic draft and quasi-neutrality in large facultatively sexual populations.
Large populations may contain numerous simultaneously segregating polymorphisms subject to natural selection. Since selection acts on individuals whose fitness depends on many loci, different loci affect each other's dynamics. This leads to stochastic fluctuations of allele frequencies above and beyond genetic drift-an effect known as genetic draft. Since recombination disrupts associations between alleles, draft is strong when recombination is rare. Here, we study a facultatively outcrossing population in a regime where the frequency of outcrossing and recombination, r, is small compared to the characteristic scale of fitness differences σ. In this regime, fit genotypes expand clonally, leading to large fluctuations in the number of recombinant offspring genotypes. The power law tail in the distribution of the latter makes it impossible to capture the dynamics of draft by an effective neutral model. Instead, we find that the fixation time of a neutral allele increases only slowly with the population size but depends sensitively on the ratio r/σ. The efficacy of selection is reduced dramatically and alleles behave "quasi-neutrally" even for Ns≫1, provided that |s| < s(c), where s(c) depends strongly on r/σ, but only weakly on population size N. In addition, the anomalous fluctuations due to draft change the spectrum of (quasi)-neutral alleles from f(ν) ∼ ν(-1), corresponding to drift, to ∼ ν(-2). Finally, draft accelerates the rate of two-step adaptations through deleterious intermediates.


Dopamine-deficient mice are severely hypoactive, adipsic, and aphagic.
Mice unable to synthesize dopamine (DA) specifically in dopaminergic neurons were created by inactivating the tyrosine hydroxylase (TH) gene then by restoring TH function in noradrenergic cells. These DA-deficient (DA-/-) mice were born at expected frequency but became hypoactive and stopped feeding a few weeks after birth. Midbrain dopaminergic neurons, their projections, and most characteristics of their target neurons in the striatum appeared normal. Within a few minutes of being injected with L-dihdroxyphenylalanine (L-DOPA), the product of TH, the DA-/- mice became more active and consumed more food than control mice. With continued administration of L-DOPA, nearly normal growth was achieved. These studies indicate that DA is essential for movement and feeding, but is not required for the development of neural circuits that control these behaviors.


A case of surgery for congenital esophagobronchial fistula accompanied by a destroyed lung.
Congenital esophagobronchial fistula (EBF) is rarely seen in adults. We report a case of EBF detected in adulthood with a destroyed lung. A 67-year-old man experienced repeated pneumonia during his childhood. Since the age of 38, he had often suffered from bloody phlegm and always had a cough and sputum during oral intake. Before cardiac surgery for atrial fibrillation and valvular disease, computed tomography (CT) detected bronchiectasis, which could cause pulmonary bleeding during heart surgery, and the patient was introduced to our hospital for lung resection. A fistula between the esophagus and the right lower lung lobe was found using CT, esophagoscopy, and esophagography. Contrast CT and angiography revealed an abnormal artery branching from the inferior phrenic artery into the lobe. As indicated by intraoperative findings, the middle and lower lobes had strongly adhered to chest wall and diaphragm, but we located the fistula easily without adhesion to the surroundings, severed it using an automatic stapler, and resected the middle and lower lobes. The symptoms disappeared immediately, and the patient was uneventfully discharged.The diagnosis of congenital EBF was established with intraoperative findings and pathological exam. The existence of pulmonary sequestration was suggested because of the long-term absence of any symptoms during his adulthood, the tract of the EBF running into the lung, not directly into the bronchus, and a septum pathologically detected in the right lower lobe. A congenital EBF should be considered for differential diagnosis in cases of limited bronchiectasis in elderly people.


Endocannabinoid system in systemic lupus erythematosus: First evidence for a deranged 2-arachidonoylglycerol metabolism.
The endocannabinoid (eCB) system plays a key role in many physiological and pathological conditions and its dysregulation has been described in several rheumatological and autoimmune diseases. Yet, its possible alteration in systemic lupus erythematosus (SLE) has never been investigated. Here, we aimed filling this gap in plasma and peripheral blood mononuclear cells (PBMCs) of patients with SLE and age- and sex- matched healthy subjects (HS). Liquid chromatography-mass spectrometry quantitation of eCB levels highlighted that plasma levels of 2-arachidonoylglycerol (2-AG) were significantly increased in SLE patients compared to HS (p = 0.0059), and among SLE patients, highest 2-AG levels were associated with a lower disease activity. No differences were found in N-arachidonoylethanolamine (AEA) and its congeners N-palmitoylethanolamine (PEA) and N-oleoylethanolamine (OEA) concentrations between the two groups. Moreover, gene expression analysis of metabolic enzymes and receptor targets of eCBs and investigation of functional activity and protein expression of selected components of eCB system disclosed a deranged 2-AG metabolism in patients with SLE. Indeed, expression and functional activity of 2-AG biosynthetic enzyme DAGL were selectively enhanced in PBMCs of SLE patients compared to HS. In conclusion, our results demonstrate, for the first time, an alteration of eCB system in SLE patients. They represents the first step toward the understanding of the role of eCB system in SLE that likely suggest DAGL and 2-AG as potential biomarkers of SLE in easily accessible blood samples. Our data provides proof-of-concept to the development of cannabis-based medicine as immune-modulating agents.


Early detection and biomarkers in pancreatic cancer.
Major advances in cancer control will be greatly aided by early detection for diagnosing and treating cancer in its preinvasive state before metastasis. Unfortunately, for pancreatic ductal adenocarcinoma (PDAC), which is the fourth leading cause of cancer-related death in the United States, effective early detection and screening are currently not available and tumors are typically diagnosed at a late stage, frequently after metastasis. Partly because of low sensitivity/specificity, existing biomarkers such as CA19-9 are not adequate as early detection markers of pancreatic cancer. Thus, a great need exists for new biomarkers for pancreatic cancer. This article focuses on recent developments in the identification of new serum protein biomarkers that are useful in the early detection of PDAC.


BACE knockout mice are healthy despite lacking the primary beta-secretase activity in brain: implications for Alzheimer's disease therapeutics.
Alzheimer's disease (AD) is a neurodegenerative disorder characterized by accumulation of amyloid plaques and neurofibrillary tangles in the brain. The major components of plaque, beta-amyloid peptides (Abetas), are produced from amyloid precursor protein (APP) by the activity of beta- and gamma-secretases. beta-secretase activity cleaves APP to define the N-terminus of the Abeta1-x peptides and, therefore, has been a long- sought therapeutic target for treatment of AD. The gene encoding a beta-secretase for beta-site APP cleaving enzyme (BACE) was identified recently. However, it was not known whether BACE was the primary beta-secretase in mammalian brain nor whether inhibition of beta-secretase might have effects in mammals that would preclude its utility as a therapeutic target. In the work described herein, we generated two lines of BACE knockout mice and characterized them for pathology, beta-secretase activity and Abeta production. These mice appeared to develop normally and showed no consistent phenotypic differences from their wild-type littermates, including overall normal tissue morphology and brain histochemistry, normal blood and urine chemistries, normal blood-cell composition, and no overt behavioral and neuromuscular effects. Brain and primary cortical cultures from BACE knockout mice showed no detectable beta-secretase activity, and primary cortical cultures from BACE knockout mice produced much less Abeta from APP. The findings that BACE is the primary beta-secretase activity in brain and that loss of beta-secretase activity produces no profound phenotypic defects with a concomitant reduction in beta-amyloid peptide clearly indicate that BACE is an excellent therapeutic target for treatment of AD.


Comparative antioxidant and anti-inflammatory effects of [6]-gingerol, [8]-gingerol, [10]-gingerol and [6]-shogaol.
Zingiber officinale Rosc. (Zingiberaceae) has been traditionally used in Ayurvedic, Chinese and Tibb-Unani herbal medicines for the treatment of various illnesses that involve inflammation and which are caused by oxidative stress. Although gingerols and shogaols are the major bioactive compounds present in Zingiber officinale, their molecular mechanisms of actions and the relationship between their structural features and the activity have not been well studied. The aim of the present study was to examine and compare the antioxidant and anti-inflammatory activities of gingerols and their natural analogues to determine their structure-activity relationship and molecular mechanisms. The in vitro activities of the compounds [6]-gingerol, [8]-gingerol, [10]-gingerol and [6]-shogaol were evaluated for scavenging of 1,1-diphenyl-2-picyrlhydrazyl (DPPH), superoxide and hydroxyl radicals, inhibition of N-formyl-methionyl-leucyl-phenylalanine (f-MLP) induced reactive oxygen species (ROS) production in human polymorphonuclear neutrophils (PMN), inhibition of lipopolysaccharide induced nitrite and prostaglandin E(2) production in RAW 264.7 cells. In the antioxidant activity assay, [6]-gingerol, [8]-gingerol, [10]-gingerol and [6]-shogaol exhibited substantial scavenging activities with IC(50) values of 26.3, 19.47, 10.47 and 8.05 microM against DPPH radical, IC(50) values of 4.05, 2.5, 1.68 and 0.85 microM against superoxide radical and IC(50) values of 4.62, 1.97, 1.35 and 0.72 microM against hydroxyl radical, respectively. The free radical scavenging activity of these compounds also enhanced with increasing concentration (P<0.05). On the other hand, all the compounds at a concentration of 6 microM have significantly inhibited (P<0.05) f-MLP-stimulated oxidative burst in PMN. In addition, production of inflammatory mediators (NO and PGE(2)) has been inhibited significantly (P<0.05) and dose-dependently. 6-Shogaol has exhibited the most potent antioxidant and anti-inflammatory properties which can be attributed to the presence of alpha,beta-unsaturated ketone moiety. The carbon chain length has also played a significant role in making 10-gingerol as the most potent among all the gingerols. This study justifies the use of dry ginger in traditional systems of medicine.


Identification and characterization of three members of the human metallocarboxypeptidase gene family.
Amino acid homology searches of the human genome revealed three members of the metallocarboxypeptidase (metallo-CP) family that had not been described in the literature in addition to the 14 known genes. One of these three, named CPA5, is present in a gene cluster with CPA1, CPA2, and CPA4 on chromosome 7. The cDNA encoding a mouse homolog of human CPA5 was isolated from a testis library and sequenced. The deduced amino acid sequence of human CPA5 has highest amino acid sequence identity (60%) to CPA1. Modeling analysis shows the overall structure to be very similar to that of other members of the A/B subfamily of metallocarboxypeptidases. The active site of CPA5 is predicted to cleave substrates with C-terminal hydrophobic residues, as do CPA1, -2, and -3. Using Northern blot analysis, CPA5 mRNA is detected in testis but not in kidney, liver, brain, or lung. In situ hybridization analysis shows that CPA5 is localized to testis germ cells. Mouse pro-CPA5 protein expressed in Sf9 cells using the baculovirus system was retained in the particulate fraction of the cells and was not secreted into the media. Pro-CPA5 was not enzymatically active toward standard CPA substrates, but after incubation with prohormone convertase 4 the resulting protein was able to cleave furylacryloyl-Gly-Leu, with 3-4-fold greater activity at pH 7.4 than at 5.6. Two additional members of the human CP gene family were also studied. Modeling analysis indicates that both contain the necessary amino acids required for enzymatic activity. The CP on chromosome 8 is predicted to have a CPA-like specificity for C-terminal hydrophobic residues and was named CPA6. The CP on chromosome 2 is predicted to cleave substrates with C-terminal acidic residues and was named CPO.


[DNA extraction and identification of Trichophyton rubrum by real-time polymerase chain reaction from direct nail scraping specimens of patients with onycomycosis].
Trichophyton rubrum is the most frequently encountered dermatophyte species causing onichomycosis. The routine diagnosis of dermatophytes depends on the direct microscopic examination (DME) and culture methods, however due to the phenotypic identification problems related to those agents, the molecular methods come into question. The aim of this study was to evaluate the diagnostic performance of real-time polymerase chain reaction (RT-PCR) for the identification of T.rubrum by comparing to DME and culture methods, from nail samples of patients with the complaints of onychomycosis. A total of 90 patients of whom 58 were male who were admitted to the dermatology outpatients clinics of our hospital with the complaints of color/shape changes in the nails and thickening of the nail, were included in the study, together with the 20 healthy volunteer subjects as controls. The nail scraping samples obtained from the patients and controls were examined with direct microscopy using 15% potassium hydroxide, dimethyl sulphoxide and chlorazole black mixture and cultivated onto Sabouraud dextrose agar with and without cycloheximide. For DNA isolation, after the disruption of nail samples with a steel tool, phenol-chloroform-isoamyl alcohol purification method were used. The amplification and demonstration of the T.rubrum DNA have been performed by using specific primers and probes following TaqMan protocol of RT-PCR (LightCycler-Roche, USA) method. Seventy-two of the patients yielded positive and 18 yielded negative results with DME. Growth of molds was detected in the cultures of 20 (27.8%) of the 72 DME positive patients and all of the isolates were identified as T.rubrum. No fungal growth was seen in the samples of 18 patients who were DME negative. In DME positive group, 67 (93%) patients were found to be positive in RT-PCR, while 8 (44.4%) patients were RT-PCR positive in DME negative group. All of the culture positive samples (n= 20) were also found positive in RT-PCR. All of the samples from the control group with healthy nails yielded negative results in DME, culture and RT-PCR methods. The performance of PCR method were compared to direct microscopy that had higher sensitivity than culture and the sensitivity, specificity, positive and negative predictive values of RTPCR assay were estimated as 93%, 56%, 89% and 67%, respectively. In conclusion RT-PCR was thought to be an efficient and rapid assay in the diagnosis of onichomycosis. Although RT-PCR seems more expensive than culture, for the centres which already have support for the molecular methods, the difference in total cost doesn't count much. In conclusion, by the use of molecular methods DNA isolation was successfully done from a relatively difficult clinical specimen, namely nail scraping, a protocole that could easily be applied in routine laboratory was established and species-level identification in a short time was accomplished in this study.


DNA adenine methylation regulates virulence gene expression in Salmonella enterica serovar Typhimurium.
Transcriptomic analyses during growth in Luria-Bertani medium were performed in strain SL1344 of Salmonella enterica serovar Typhimurium and in two isogenic derivatives lacking Dam methylase. More genes were repressed than were activated by Dam methylation (139 versus 37). Key genes that were differentially regulated by Dam methylation were verified independently. The largest classes of Dam-repressed genes included genes belonging to the SOS regulon, as previously described in Escherichia coli, and genes of the SOS-inducible Salmonella prophages ST64B, Gifsy-1, and Fels-2. Dam-dependent virulence-related genes were also identified. Invasion genes in pathogenicity island SPI-1 were activated by Dam methylation, while the fimbrial operon std was repressed by Dam methylation. Certain flagellar genes were repressed by Dam methylation, and Dam(-) mutants of S. enterica showed reduced motility. Altered expression patterns in the absence of Dam methylation were also found for the chemotaxis genes cheR (repressed by Dam) and STM3216 (activated by Dam) and for the Braun lipoprotein gene, lppB (activated by Dam). The requirement for DNA adenine methylation in the regulation of specific virulence genes suggests that certain defects of Salmonella Dam(-) mutants in the mouse model may be caused by altered patterns of gene expression.


Evidence Implicating Immunological Host Effects in the Efficacy of Metronomic Low-Dose Chemotherapy.
Conventional chemotherapy drugs administered at a maximum tolerated dose (MTD) remains the backbone for treating most cancers. Low-dose metronomic (LDM) chemotherapy, which utilizes lower, less toxic, doses given on a close regular basis over prolonged periods, is an alternative and better tolerated potential strategy to improve chemotherapy. LDM chemotherapy has been evaluated preclinically and clinically and has shown therapeutic benefit, in both early and advanced stage metastatic disease, especially when used as a maintenance therapy. However, knowledge about the antitumor mechanisms by which LDM chemotherapy acts remain limited. Here we characterized the effects of LDM and MTD capecitabine therapy on tumor and host cells using high-throughput systems approaches involving mass spectrometry flow cytometry and automated cell imaging followed by in vivo analyses of such therapies. An increase in myeloid and T regulatory cells and a decrease in NK and T cytotoxic cells were found in MTD-capecitabine-treated tumors compared with LDM-capecitbine-treated tumors. Plasma from MTD capecitabine-treated mice induced a more tumorigenic and metastatic profile in both breast and colon carcinoma cells than plasma from mice treated with LDM capecitabine. These results correlated, in part, with in vivo studies using models of human or mouse advanced metastatic disease, where the therapeutic advantage of MTD capecitabine was limited despite a substantial initial antitumor activity found in the primary tumor setting. Overall these results implicate a possible contribution of immunologic host effects in accounting for the therapeutic limitations of MTD compared with LDM capecitabine. Cancer Res; 76(20); 5983-93. ©2016 AACR.


The use of delta-tocotrienol and lovastatin for anti-osteoporotic therapy.
Statins are competitive inhibitors of HMGCoA reductase and are commonly used as antihypercholesterolemic agents. Experimental studies clearly demonstrate the beneficial effects of statins on bone. Tocotrienols have also been shown to have anti-osteoporotic effects on the skeletal system. This study was conducted to observe the effect of a combination of delta-tocotrienol and lovastatin on structural bone histomorphometry and bone biomechanical strength in a postmenopausal rat model at clinically tolerable doses, and to compare it with the effect of delta-tocotrienol or lovastatin. Forty-eight female Sprague Dawley rats were randomly divided into six groups: baseline control; sham-operated control; ovariectomised control; ovariectomised+11mg/kg lovastatin; ovariectomised+60mg/kg delta-tocotrienol and ovariectomised+60mg/kg delta-tocotrienol+11mg/kg lovastatin. These treatments were given via oral gavage daily for eight weeks. After sacrificing the rats, the left and right femurs were dissected and processed for bone histomorphometric analysis and a bone biomechanical test, respectively. Delta-tocotrienol in combination with lovastatin significantly improved the trabecular volume, trabecular number, trabecular thickness and trabecular separation; and it significantly increased bone strength in oestrogen-deficient rats. Delta-tocotrienol alone enhanced bone formation and maintained bone strength in ovariectomised rats. Delta-tocotrienol plus lovastatin treatment promoted better trabecular volume and trabecular number and received higher load than delta-tocotrienol alone. Lovastatin alone was not effective. Thus, the combination of delta-tocotrienol and lovastatin has the potential to be used for anti-osteoporotic therapy in postmenopausal women.


Improved dietary variety and adequacy but lower dietary moderation with acculturation in Chinese women in the United States.
Acculturation is associated with increased chronic disease risk among Asian Americans, but its association with different aspects of diet quality remains unclear. Associations of acculturation with diet quality were examined in a convenience sample of 243 Chinese participants in a study of diet and mammographic density in the Philadelphia region between January 2002 to May 2003. An acculturation index was created based on self-reported English proficiency and within- and cross-ethnicity social interactions. Diet Quality Index-International (DQI-I) scores were based on responses to an 88-item food frequency questionnaire. Odds ratios (ORs) for falling into a higher vs lower quartile for DQI-I and its components (ie, variety, adequacy, moderation, balance) were estimated with logistic regression analysis for polytomous outcomes. In the sample, mean age was 53.2 (standard deviation=10.5) years, body mass index (calculated as kg/m2) was 24.1 (standard deviation=3.5), and acculturation was significantly associated with improved dietary variety (OR: 2.4; 95% confidence interval [CI]: 1.5 to 3.8) and adequacy (OR: 1.6; 95% CI: 1.0 to 2.6) and lower dietary moderation (OR: 0.6; 95% CI 0.4 to 0.9), but these associations were evident only among women with less than a high school education. Acculturation and education were not associated with overall diet quality or balance. Although an association of less dietary moderation with acculturation suggests the likely importance of acculturation-related dietary change to chronic disease risk, these findings highlight the need for flexible dietary interventions among immigrant populations to discourage the adoption of some new dietary habits, while encouraging the retention of other, traditional ones.


In situ lipid dip-pen nanolithography under water.
Lipids form the structural and functional basis of biological membranes, and methods for studying their self-organization in well-defined nano- and micro-scale model systems can provide insights into biology. Using lipids as an ink for dip-pen nanolithography (lipid DPN) permits the rapid nanostructuring of multicomponent model lipid membrane systems, but this technique has so far been limited to air. Here we demonstrate that lipid DPN can be carried out under water with single tips or parallel arrays. Using the same tip for deposition and imaging in aqueous solution permits imaging of self-spreading lipid bilayer spots in situ and quantification of the nanoscale spreading kinetics in real time by means of lateral-force microscopy. Furthermore, using fluorophore-labeled phospholipids, we directly observed, by confocal laser scanning microscopy, a two-phase (oil in water) meniscus formed around the contact point between the DPN tip and surface, gaining insights into the mechanisms of the ink transport. The methods described here provide a new tool and environment for high-resolution studies of lipid nanodynamics and molecular printing processes in general.


Supplements and sports.
Use of performance-enhancing supplements occurs at all levels of sports, from professional athletes to junior high school students. Although some supplements do enhance athletic performance, many have no proven benefits and have serious adverse effects. Anabolic steroids and ephedrine have life-threatening adverse effects and are prohibited by the International Olympic Committee and the National Collegiate Athletic Association for use in competition. Blood transfusions, androstenedione, and dehydroepiandrosterone are also prohibited in competition. Caffeine, creatine, and sodium bicarbonate have been shown to enhance performance in certain contexts and have few adverse effects. No performance benefit has been shown with amino acids, beta-hydroxy-beta-methylbutyrate, chromium, human growth hormone, and iron. Carbohydrate-electrolyte beverages have no serious adverse effects and can aid performance when used for fluid replacement. Given the widespread use of performance-enhancing supplements, physicians should be prepared to counsel athletes of all ages about their effectiveness, safety, and legality.


Yes-associated protein impacts adherens junction assembly through regulating actin cytoskeleton organization.
The Hippo pathway effector Yes-associated protein (YAP) regulates liver size by promoting cell proliferation and inhibiting apoptosis. However, recent in vivo studies suggest that YAP has important cellular functions other than controlling proliferation and apoptosis. Transgenic YAP expression in mouse hepatocytes results in severe jaundice. A possible explanation for the jaundice could be defects in adherens junctions that prevent bile from leaking into the blood stream. Indeed, immunostaining of E-cadherin and electron microscopic examination of bile canaliculi of Yap transgenic livers revealed abnormal adherens junction structures. Using primary hepatocytes from Yap transgenic livers and Yap knockout livers, we found that YAP antagonizes E-cadherin-mediated cell-cell junction assembly by regulating the cellular actin architecture, including its mechanical properties (elasticity and cortical tension). Mechanistically, we found that YAP promoted contractile actin structure formation by upregulating nonmuscle myosin light chain expression and cellular ATP generation. Thus, by modulating actomyosin organization, YAP may influence many actomyosin-dependent cellular characteristics, including adhesion, membrane protrusion, spreading, morphology, and cortical tension and elasticity, which in turn determine cell differentiation and tissue morphogenesis.


The Role of Dysfunctional Cognitions in Patients With Chronic Tinnitus.
The present study investigates the role of dysfunctional cognitions in patients with chronic tinnitus. To explore different dimensions of tinnitus-related thoughts, a 22-item self-report measure, the "Tinnitus Cognitions Scale" (T-Cog), is presented. Furthermore, dysfunctional cognitions are examined as a possible mediator of the relation between tinnitus distress and depression. The present study analyzes the cross-sectional data of 373 patients with chronic tinnitus. Parallel analysis and principal axis factoring are used to identify the factor structure of the T-Cog. Assumed mediating effects are tested using the asymptotic and resampling procedure. Factor analysis reveals two factors interpreted as "tinnitus-related catastrophic thinking" and "tinnitus-related avoidance cognitions." Internal consistency is sufficient with a Cronbach's α of 0.88 for the total scale and 0.74 and 0.87 for the subscales. The authors find high associations between the T-Cog and other measures of tinnitus distress, depression, anxiety, and tinnitus acceptance, indicating convergent validity. With the exception of neuroticism, low correlations with personality factors are found, indicating discriminant validity. Patients with moderate or severe tinnitus distress report significantly higher scores of dysfunctional cognitions than patients with mild tinnitus distress. Tinnitus-related catastrophic thinking and tinnitus-related avoidance cognitions partially mediate the relation between tinnitus distress and depression. Dysfunctional cognitions can play an important role in the degree of tinnitus distress. Catastrophic and avoidant thoughts contribute to the explanation of depression among tinnitus patients. The T-Cog is a reliable and valid questionnaire for the assessment of different dimensions of cognitions. Its use could provide information for identifying tinnitus patients who are particularly suitable for cognitive-behavioral therapy.


Enhanced-affinity murine T-cell receptors for tumor/self-antigens can be safe in gene therapy despite surpassing the threshold for thymic selection.
Many of the most promising tumor antigens for T-cell-based cancer immunotherapies are unmodified self-antigens. Unfortunately, the avidity of T cells specific for these antigens is limited by central tolerance during T-cell development in the thymus, resulting in decreased anti-tumor efficacy of these T cells. One approach to overcoming this obstacle is to mutate T-cell receptor (TCR) genes from naturally occurring T cells to enhance the affinity for the target antigen. These enhanced-affinity TCRs can then be developed for use in TCR gene therapy. Although TCRs with significantly enhanced affinity have been generated using this approach, it is not clear whether these TCRs, which bypass the affinity limits imposed by negative selection, remain unresponsive to the low levels of self-antigen generally expressed by some normal tissues. Here we show that 2 variants of a high-affinity WT1-specific TCR with enhanced affinity for WT1 are safe and do not mediate autoimmune tissue infiltration or damage when transduced into peripheral CD8 T cells and transferred in vivo. However, if expressed in developing T cells and subjected to thymic selection, the same enhanced-affinity TCRs signal tolerance mechanisms in the thymus, resulting in T cells with attenuated antigen sensitivity in the periphery.


Breastfeeding may improve nocturnal sleep and reduce infantile colic: potential role of breast milk melatonin.
Melatonin is secreted during the night in adults but not in infants. It has a hypnotic effect as well as a relaxing effect on the smooth muscle of the gastrointestinal tract. It is plausible that breast milk, which consists of melatonin, may have an effect on improving infants' sleep and reducing infantile colic. Our first goal was to assess the differences in the prevalence and severity of infantile colic and nocturnal sleep between breast-fed infants and supplement-fed infants. The second was to characterize the profile of melatonin secretion in human breast milk compared to artificial formulas. Ninety-four mothers of healthy 2 to 4-month-old infants filled a questionnaire regarding irritability/potential infantile colic and sleep characteristics. For the second part, we measured melatonin levels in breast milk of five women every 2 h during 24 h and in three samples of commonly used artificial formulas. Exclusively breast-fed infants had a significantly lower incidence of colic attacks (p = 0.04), lower severity of irritability attacks (p = 0.03), and a trend for longer nocturnal sleep duration (p = 0.06). Melatonin in human milk showed a clear circadian curve and was unmeasurable in all artificial milks. Exclusive breastfeeding is associated with reduced irritability/colic and a tendency toward longer nocturnal sleep. Breast milk (nocturnal) consists of substantial melatonin levels, whereas artificial formulas do not. We speculate that melatonin which is supplied to the infant via breast milk plays a role in improving sleep and reducing colic in breast-fed infants compared to formula-fed ones.


Orbitofrontal cortex and social behavior: integrating self-monitoring and emotion-cognition interactions.
The role of the orbitofrontal cortex in social behavior remains a puzzle. Various theories of the social functions of the orbitofrontal cortex focus on the role of this area in either emotional processing or its involvement in online monitoring of behavior (i.e., self-monitoring). The present research attempts to integrate these two theories by examining whether improving the self-monitoring of patients with orbitofrontal damage is associated with the generation of emotions needed to guide interpersonal behavior. Patients with orbitofrontal damage, patients with lateral prefrontal damage, and healthy controls took part in an interpersonal task. After completing the task, participants' self-monitoring was increased by showing them a videotape of their task performance. In comparison to healthy controls and patients with lateral prefrontal damage, orbitofrontal damage was associated with objectively inappropriate social behavior. Although patients with orbitofrontal damage were aware of social norms of intimacy, they were unaware that their task performance violated these norms. The embarrassment typically associated with inappropriate social behavior was elicited in these patients only after their self-monitoring increased from viewing their videotaped performance. These findings suggest that damage to the orbitofrontal cortex impairs self-insight that may preclude the generation of helpful emotional information. The results highlight the role of the orbitofrontal cortex in the interplay of self-monitoring and emotional processing and suggest avenues for neurorehabilitation of patients with social deficits subsequent to orbitofrontal damage.


Novel GMP-compatible protocol employing an allogeneic B cell bank for clonal expansion of allospecific natural regulatory T cells.
The adoptive transfer of natural regulatory T cells (nTreg) is a new option to reshape undesired immune reactivity in autoimmunity and transplantation toward "tolerance." The first clinical trials using adoptive transfer of polyclonal nTreg demonstrated safety and hints of efficacy. However, the low frequencies of antigen-specific cells among the pool of polyclonal nTreg and their broad antigen nonspecific suppression are limitations of this approach regarding efficacy and safety. Recently, the isolation and expansion of (allo)antigen-specific nTreg have successfully been achieved by using Treg-specific activation markers but the yield is relatively low. Here, we describe a novel good manufacturing practice (GMP)-compatible expansion protocol of alloantigen-specific nTreg based on the stimulation of nTreg by allogeneic activated B cells. Their functionality and specificity are superior compared to polyclonal nTreg both in vitro and in vivo. Employing an allogeneic B cell bank, designed to cover the majority of HLA types, allows fast GMP-compliant manufacturing for donor-specific nTreg for clinical application in organ and stem cell transplantation. TCR repertoire analyses by next generation sequencing revealed impressive expansion by several log-steps of even very low-abundance alloantigen-specific nTreg clones. This novel method offers a simple approach for expanding antigen-specific nTreg and is characterized by high replicability and easy transferability to full GMP standards.


Pathogenesis of carbohydrate-induced hypertriglyceridemia using HepG2 cells as a model system.
This study compares the effects of glucose and fatty acid on hepatic lipid synthesis and apolipoprotein (apo) B secretion. To do so, varying concentrations of either glucose or oleic acid were added to the medium in which HepG2 cells were being incubated. Intracellular triacylglycerol and cholesteryl ester synthesis and secretion were measured by addition of radioisotopic tracers and by determination of mass, whereas apo B concentration in the medium was measured by a specific enzyme-linked immunosorbent assay. The data indicate that increasing concentrations of glucose in the medium resulted in increased synthesis of triacylglycerol within the cell and increased secretion of triacylglycerol into the medium. Apo B secretion into the medium, however, did not change, and intracellular synthesis and secretion of cholesteryl ester did not change as well. By contrast, addition of oleic acid to the medium resulted in increased synthesis and secretion of both cholesteryl ester and triacylglycerol, and this was associated with increased secretion of apo B into the medium. Thus, a carbohydrate load resulted in secretion of normal numbers of triacylglycerol-enriched apo B particles by this hepatocyte cell line, whereas a fatty acid load led to the secretion of increased numbers of apo B particles, which were essentially normal in composition.


Comparison of effects of static, proprioceptive neuromuscular facilitation and Mulligan stretching on hip flexion range of motion: a randomized controlled trial.
The aim of this study was to compare the effects of static stretching, proprioceptive neuromuscular facilitation (PNF) stretching and Mulligan technique on hip flexion range of motion (ROM) in subjects with bilateral hamstring tightness. A total of 40 students (mean age: 21.5±1.3 years, mean body height: 172.8±8.2 cm, mean body mass index: 21.9±3.0 kg · m(-2)) with bilateral hamstring tightness were enrolled in this randomized trial, of whom 26 completed the study. Subjects were divided into 4 groups performing (I) typical static stretching, (II) PNF stretching, (III) Mulligan traction straight leg raise (TSLR) technique, (IV) no intervention. Hip flexion ROM was measured using a digital goniometer with the passive straight leg raise test before and after 4 weeks by two physiotherapists blinded to the groups. 52 extremities of 26 subjects were analyzed. Hip flexion ROM increased in all three intervention groups (p<0.05) but not in the no-intervention group after 4 weeks. A statistically significant change in initial-final assessment differences of hip flexion ROM was found between groups (p<0.001) in favour of PNF stretching and Mulligan TSLR technique in comparison to typical static stretching (p=0.016 and p=0.02, respectively). No significant difference was found between Mulligan TSLR technique and PNF stretching (p=0.920). The initial-final assessment difference of hip flexion ROM was similar in typical static stretching and no intervention (p=0.491). A 4-week stretching intervention is beneficial for increasing hip flexion ROM in bilateral hamstring tightness. However, PNF stretching and Mulligan TSLR technique are superior to typical static stretching. These two interventions can be alternatively used for stretching in hamstring tightness.


Regulation of human T lymphocyte coactivation with an alpha4 integrin antagonist peptide.
The cyclic hexapeptide CWLDVC (TBC 772) is an antagonist of alpha4 integrins and a potent inhibitor of lymphocyte interactions with fibronectin, vascular cell adhesion molecule-1, and muscosal vascular addressin cell adhesion molecule-1 (MAdCAM-1). As such, peptide TBC 772 effectively inhibits the activation of freshly isolated human T lymphocytes stimulated with purified vascular cell adhesion molecule-1 coimmobilized with anti-CD3 mAb. The influence of peptide binding on distinct sites of the alpha4beta1 complex was determined by flow cytometry and cellular adhesion assays employing a panel of mAbs. Binding of the alpha4-specific mAb L25 and the beta1-specific mAb 33B6 was not altered by the peptide; however, binding of mAb 19H8, which is specific for a combinatorial epitope of alpha4beta1, was dramatically inhibited. Treatment of lymphocytes with the peptide caused an increase in a ligand-induced epitope on beta1 integrin defined by mAb 15/7. In T cell activation studies using coimmobilized anti-CD3 mAb and the anti-integrin mAbs, the peptide had broader inhibitory activity, suppressing costimulation induced by all the integrin mAbs. The peptide was not generally toxic and was integrin selective in its suppressive activity, as coactivation by ligation of CD3 in conjunction with CD28 or CD26 was not affected. These results suggest that the antagonist peptide CWLDVC can effectively neutralize integrin coactivation systems by a mechanism independent of competitive binding.


Divergent functions of the Rho GTPases Rac1 and Cdc42 in podocyte injury.
Podocytes are highly specialized epithelial cells with complex actin cytoskeletal architecture crucial for maintenance of the glomerular filtration barrier. The mammalian Rho GTPases Rac1 and Cdc42 are molecular switches that control many cellular processes, but are best known for their roles in the regulation of actin cytoskeleton dynamics. Here, we employed podocyte-specific Cre-lox technology and found that mice with deletion of Rac1 display normal podocyte morphology without glomerular dysfunction well into adulthood. Using the protamine sulfate model of acute podocyte injury, podocyte-specific deletion of Rac1 prevented foot process effacement. In a long-term model of chronic hypertensive glomerular damage, however, loss of Rac1 led to an exacerbation of albuminuria and glomerulosclerosis. In contrast, mice with podocyte-specific deletion of Cdc42 had severe proteinuria, podocyte foot process effacement, and glomerulosclerosis beginning as early as 10 days of age. In addition, slit diaphragm proteins nephrin and podocin were redistributed, and cofilin was dephosphorylated. Cdc42 is necessary for the maintenance of podocyte structure and function, but Rac1 is entirely dispensable in physiological steady state. However, Rac1 has either beneficial or deleterious effects depending on the context of podocyte impairment. Thus, our study highlights the divergent roles of Rac1 and Cdc42 function in podocyte maintenance and injury.


Reaction mechanisms for the CO oxidation on Au/CeO(2) catalysts: activity of substitutional Au(3+)/Au(+) cations and deactivation of supported Au(+) adatoms.
Density functional theory calculations that account for the on-site Coulomb interaction via a Hubbard term (DFT+U) reveal the mechanisms for the oxidation of CO catalyzed by isolated Au atoms as well as small clusters in Au/CeO(2) catalysts. Ceria (111) surfaces containing positively charged Au ions, either as supported Au(+) adatoms or as substitutional Au(3+) ions, are shown to activate molecular CO and to catalyze its oxidation to CO(2). In the case of supported single Au(+) adatoms, the limiting rate for the CO oxidation is determined by the adsorbate spillover from the adatom to the oxide support. The reaction then proceeds with the CO oxidation via lattice oxygen and O vacancy formation. These vacancies are shown to readily attract the supported Au(+) adatoms and to turn them into negatively charged Au(delta-) adspecies that deactivate the catalyst, preventing further CO adsorption. Au(3+) ions dispersed into the ceria lattice as substitutional point defects can instead sustain a full catalytic cycle consisting of three individual steps maintaining their activity along the reaction process: Au cations in Au(x)Ce(1-x)O(2) systems promote multiple oxidations of CO without any activation energy via formation of surface O vacancies. Molecular oxygen adsorbs at these vacancies and forms O adspecies that then catalyze the oxidation of molecular CO, closing the catalytic cycle and recovering the stoichiometric Au(x)Ce(1-x)O(2) system. The interplay between the reversible Ce(4+)/Ce(3+) and Au(3+)/Au(+) reductions underpins the high catalytic activity of dispersed Au atoms into the ceria substrate. It is shown that the positive oxidation state of the substitutional Au ions is retained along the catalytic cycle, thus preventing the deactivation of Au(x)Ce(1-x)O(2) catalysts in operation conditions. Finally, although a single Au(+) adatom bound to an O vacancy is shown to deactivate during CO oxidation, the calculations predict that the reactivity of gold nanoparticles nucleated at O vacancies can be recovered for cluster sizes as small as Au(2).


Molecular biology of flaviviruses.
Flaviviruses are enveloped viruses with a single-stranded, 10.7kb positive-sense RNA genome. The genomic RNA, which has a 5' cap but no poly(A) tail, is translated as a single polyprotein that is then cleaved into three structural proteins and seven non-structural (NS) proteins by both viral and host proteases. The NS proteins include an RNA-dependent RNA polymerase (NS5), a helicase/protease (NS3), and other proteins that form part of the viral replication complex. Sequences and structures in the 5' and 3' untranslated regions (UTR) and capsid gene, including the cyclization sequences, the upstream AUG region, and the terminal 3' stem-loop, regulate translation, RNA synthesis and viral replication. We have also found that an RNA hairpin structure in the capsid coding region (cHP) influences start codon selection and viral replication of the flavivirus dengue virus (DENV). Peptide-conjugated phosphorodiamidate morpholino oligomers (P-PMOs) were used to further dissect the role of conserved regions of the 5' and 3' UTRs; several P-PMOs were shown to specifically inhibit DENV translation and/or RNA synthesis and, hence, are potentially useful as antiviral agents. Regarding the mechanism of DENV translation, we have shown that DENV undergoes canonical cap-dependent translation initiation as well as a non-canonical mechanism when cap-dependent translation is suppressed. Although much remains to be elucidated about the molecular biology of flavivirus infection, progress is being made towards defining the cis and trans factors that regulate flavivirus translation and replication.


DNA damage and nonhomologous end joining in excitotoxicity: neuroprotective role of DNA-PKcs in kainic acid-induced seizures.
DNA repair plays a critical, but imprecisely defined role in excitotoxic injury and neuronal survival throughout adulthood. We utilized an excitotoxic injury model to compare the location and phenotype of degenerating neurons in mice (strain 129-C57BL) deficient in the catalytic subunit of the DNA-dependent protein kinase (DNA-PKcs), an enzyme required for nonhomologous end joining (NHEJ). Brains from untreated adult heterozygous and DNA-PKcs null mice displayed comparable cytoarchitecture and undetectable levels of cell death. By day 1, and extending through 4 days following kainic acid-induced seizures, brains from DNA-PKcs null mice showed widespread neurodegeneration that encompassed the entire hippocampal CA1-CA3 pyramidal cell layer, entorhinal cortex, and lateral septum, with relative sparing of the dentate gyrus granule cell layer and hilus, as judged by toluidine blue, Fluoro-Jade B, and terminal dUTP nick end labeling staining. In contrast, seizure-related neurodegeneration in heterozygous littermates was limited to the CA3 region of the hippocampus. NeuN and calbindin staining revealed a selective decrease in the number and density of NeuN-positive neurons in the pyramidal layers of degenerating regions in both heterozygous and DNA-PKcs null mice. To elucidate the mechanisms leading to cell death, we examined an involvement of the p53 pathway, known to be induced by DNA damage. Addition of pifithrin-alpha, a p53 inhibitor, or expression of a dominant-negative p53 rescued neurons from kainate-induced excitotoxic cell death in primary cortical cultures derived from wildtype, DNA-PKcs heterozygous, or DNA-PKcs null neonatal mice. Moreover, pifithrin-alpha prevented kainate-induced loss of mitochondrial membrane potential, dendrite degeneration, and cell death. Results suggest that NHEJ plays a neuroprotective role in excitotoxicity, within the perforant, Schaffer collateral, hippocampal-septal, and temperoammonic pathways, in part by repairing DNA damage that would otherwise result in activation of a p53-dependent apoptotic cascade.


Metabolic syndrome: what is it and what are the implications?
Obesity and overweight are linked with a cluster of metabolic and vascular disorders that have been termed the metabolic syndrome. Although there is not yet a universally-accepted set of diagnostic criteria, most expert groups agree that the syndrome is characterised by impaired insulin sensitivity and hyperglycaemia, dyslipidaemia (elevated blood triacyglycerols with depressed HDL-cholesterol), abdominal obesity and hypertension. Based on existing published criteria estimates suggest that the syndrome affects a substantial percentage of the middle-aged and elderly populations of most European countries (10-20%) and confers increased risk of type 2 diabetes (2-8.8-fold) and CVD (1.5-6-fold), as well as having a marked effect on morbidity. Although the pathophysiology is incompletely understood, insulin resistance and abdominal obesity are central to subsequent abnormalities in circulating glucose and lipoproteins, and vascular function that lead to type 2 diabetes, atherosclerosis and CVD. The link between metabolic syndrome, type 2 diabetes and CVD, as well as inability to reverse the present rising rates of obesity, will lead to economically-unsustainable costs of health care in the next 10-20 years. Preventative strategies for metabolic syndrome are required to slow rates of progression and to reduce dependence on costly medical management. A notable development is recent evidence that shows that diet and exercise are more effective than drug treatment in preventing the development of type-2 diabetes in high-risk individuals. The LIPGENE project will investigate dietary fat quality as a strategy for the prevention of metabolic syndrome and identify food chain approaches that can support consumer attempts to alter their dietary patterns.


Drug utilization patterns in the emergency department: A retrospective study.
The aim of this study was to assess the prescribing trends and costs of drugs in the emergency department (ED) at Sultan Qaboos University Hospital (SQUH), a tertiary care hospital, in Muscat, the Sultanate of Oman. This was a retrospective cross-sectional study of all patients (n = 300) who attended the ED at SQUH in May 2012. Analyses were performed using descriptive and univariate statistics. The average age of patients was 34 ± 19 years. The average number of drugs prescribed per patients was 3.2 ± 1.9 and the majority of the patients (n = 78; 26%) received two drugs. The most common route of drug administration was the oral route (n = 481; 51%) followed by parenterally (n = 357; 38%). Non-steroidal anti-inflammatory drugs (NSAIDs) were the most commonly prescribed class of drugs (38%) followed by the gastro-intestinal tract drugs (19%) and central nervous system drugs (13%). The average cost per prescription was 242 ± 632 US$. Morphine had the highest cost (1885 US$) followed by cefuroxime (1404 US$) and filgrastim (939 US$) over the 1-month period. There was a significant positive correlation between hospital cost and age (P < 0.001), duration of stay at the ED (P = 0.008) and emergency types (P < 0.001). NSAIDs were the most frequent class of drugs administered to patients. Highest number of drugs was prescribed for cardiovascular diseases followed by respiratory and gastrointestinal diseases. Anti-infective drugs cost was the highest among all other classes. The results of the present study are attempts to highlight the importance of strategies that have to be implemented to optimize medication use at the ED.


Codeposition of organic carbon and arsenic in Bengal Delta aquifers.
We present data showing that arsenic (As) was codeposited with organic carbon (OC) in Bengal Delta sediments as As and OC concentrations are highly (p < 0.001) positively correlated in core profiles collected from widely dispersed geographical sites with different sedimentary depositional histories. Analysis of modern day depositional environments revealed that the As-OC correlations observed in cores are due to As retention and high OC inputs in vegetated zones of the deltaic environment. We hypothesize that elevated concentrations of As occur in vegetated wetland sediments due to concentration and retention of arsenate in aerated root zones and animal burrows where copious iron(III) oxides are deposited. On burial of the sediment, degradation of organic carbon from plant and animal biomass detritus provides the reducing conditions to dissolve iron(III) oxides and release arsenite into the porewater. As tubewell abstracted aquifer water is an invaluable resource on which much of Southeast Asia is now dependent, this increased understanding of the processes responsible for As buildup and release will identify, through knowledge of the palaeosedimentary environment, which sediments are at most risk of having high arsenic concentrations in porewater. Our data allow the development of a new unifying hypothesis of how As is mobilized into groundwaters in river flood plains and deltas of Southeast Asia, namely that in these highly biologically productive environments, As and OC are codeposited, and the codeposited OC drives As release from the sediments.


Protection of mice against lethal challenge with herpes simplex virus by vaccination with an adenovirus vector expressing HSV glycoprotein B.
Increasing attention has been focused on the use of recombinant mammalian viruses as potential vaccines. Recombinant human adenoviruses are one of the more promising vaccine vectors because they can be easily constructed and because live adenovirus vaccines have been administered orally to large numbers of military recruits without adverse reactions. In order to examine the efficacy of human adenoviruses as vaccines we have studied the immunity induced by a recombinant adenovirus vector, AdgB2, which induces high level expression of herpes simplex virus (HSV) glycoprotein B (gB) in human and murine cells. Mice inoculated with AdgB2 produced antibodies specific for gB which neutralized HSV in the presence of complement. Although mice inoculated with AdgB2 showed no ill-effects after AdgB2 inoculation and we were unable to detect replication of human adenoviruses in mice, the mice were protected from a lethal challenge with HSV after a single inoculation with AdgB2.


A common 16p11.2 inversion underlies the joint susceptibility to asthma and obesity.
The prevalence of asthma and obesity is increasing worldwide, and obesity is a well-documented risk factor for asthma. The mechanisms underlying this association and parallel time trends remain largely unknown but genetic factors may be involved. Here, we report on a common ~0.45 Mb genomic inversion at 16p11.2 that can be accurately genotyped via SNP array data. We show that the inversion allele protects against the joint occurrence of asthma and obesity in five large independent studies (combined sample size of 317 cases and 543 controls drawn from a total of 5,809 samples; combined OR = 0.48, p = 5.5 × 10(-6)). Allele frequencies show remarkable worldwide population stratification, ranging from 10% in East Africa to 49% in Northern Europe, consistent with discordant and extreme genetic drifts or adaptive selections after human migration out of Africa. Inversion alleles strongly correlate with expression levels of neighboring genes, especially TUFM (p = 3.0 × 10(-40)) that encodes a mitochondrial protein regulator of energy balance and inhibitor of type 1 interferon, and other candidates for asthma (IL27) and obesity (APOB48R and SH2B1). Therefore, by affecting gene expression, the ~0.45 Mb 16p11.2 inversion provides a genetic basis for the joint susceptibility to asthma and obesity, with a population attributable risk of 39.7%. Differential mitochondrial function and basal energy balance of inversion alleles might also underlie the potential selection signature that led to their uneven distribution in world populations.


Facts and hopes on RAS inhibitors and cancer immunotherapy.
While the past decade has seen great strides in the development of immunotherapies that reactivate the immune system against tumors, there have also been major advances in the discovery of drugs blocking oncogenic drivers of cancer growth. However, there has been very little progress in combining immunotherapies with drugs that target oncogenic driver pathways. Some of the most important oncogenes in human cancer encode RAS family proteins; although these have proven challenging to target, recently drugs have been approved that inhibit a specific mutant form of KRAS, G12C. These have improved the treatment of lung cancer patients harboring this mutation but development of acquired drug resistance after initial responses has limited the impact on overall survival. Due to the immunosuppressive nature of the signaling network controlled by oncogenic KRAS, targeted KRAS G12C inhibition can indirectly affect anti-tumor immunity, and does so without compromising the critical role of normal RAS proteins in immune cells. This serves as a rationale for combination with immune checkpoint blockade, which can provide additional combinatorial therapeutic benefit in some pre-clinical cancer models. However, in clinical trials, combination of KRAS G12C inhibitors with PD-(L)1 blockade has yet to show improved outcome, in part due to treatment toxicities. A greater understanding of how oncogenic KRAS drives immune evasion and how mutant specific KRAS inhibition impacts the tumor microenvironment can lead to novel approaches to combining RAS inhibition with immunotherapies.


Associations between maternal exposure to incense burning and blood pressure during pregnancy.
Incense burning is a popular practice in Asian and Arabic countries. Previous studies show that incense burning was associated with increased risks of adverse outcomes among non-pregnant population. However, very few studies explored its health effects among pregnant women, who are more susceptible to environmental stressor. We aimed to examine the association between incense burning at home and hypertensive disorders as well as blood pressure levels during pregnancy, using data from 10,563 pregnant women recruited in Born in Guangzhou Cohort Study, China between January 2013 and December 2015. Information on frequency and duration of exposure to incense burning were collected at early and late pregnancy using questionnaire. Data on outcome variables, including hypertensive disorders diagnosis and blood pressure levels at the final antenatal visit before delivery, were extracted from medical records. We used Poisson regression model and general linear model to examine the associations between incense exposure and the outcomes. We found incense use at early pregnancy was not significantly associated with outcomes. Pregnant women who frequently smelled the incense burning at late pregnancy was associated with higher risk of hypertensive disorders (relative risk, 1.84; 95% confidence interval, 1.14-2.98) and higher levels of blood pressure (1.6mmHg increase of systolic blood pressure; 95% confidence interval, 0.4-2.8mmHg) before delivery, compared to those did not burn incense. These associations tended to more evident among women without active and passive smoking. We did not observe significant dose-response relationship between exposure duration and the risk of hypertensive disorders. We firstly reported exposure to incense burning was associated with the risk of hypertensive disorders and blood pressure levels during pregnancy. Given hypertensive disorders in pregnancy are well-established risk factors for a variety of adverse outcomes and the incense burning is a modifiable factor, our finding may have important public health significance.


Chronic ACE inhibition reduces intimal hyperplasia in experimental vein grafts.
Intimal hyperplasia is an important factor in the pathophysiology of vein graft failure. Local renin-angiotensin systems recently have been shown to modulate the development of intimal hyperplasia in arteries after intimal injury. The effect of chronic angiotensin-converting enzyme (ACE) inhibition on the development of intimal hyperplasia in experimental vein grafts was examined in this study. Ten New Zealand White rabbits received 10 mg/kg of captopril daily in their drinking water. One week later the right carotid artery was divided and bypassed with the reversed right external jugular vein in these rabbits and in 10 matched controls. Captopril was continued for 28 days after operation, when all the grafts were harvested. Five grafts from each group were perfusion fixed, and the intimal thickness in the proximal, middle, and distal segments was determined. Rings from the remaining grafts (n = 20 in each group) were studied in vitro under isometric tension, and their responses to norepinephrine (NE), histamine (HIST), serotonin (5-HT), angiotensin I (AI), and angiotensin II (AII) was measured. The intimal thickness of the proximal, middle, and distal segments of the captopril-treated grafts were significantly less than controls, being reduced in all segments by approximately 40% (p less than 0.0001). With regard to vasoreactivity, the captopril-treated grafts were hypersensitive to 5-HT (control ED50 5.5 +/- 0.5 X 10(-7) mol/L vs. captopril-treated 1.1 +/- 0.2 X 10(-6) mol/L; p less than 0.005) although the maximal response was significantly reduced (control 1.6 +/- 0.3 g vs. captopril-treated 0.8 +/- 0.1 g; p less than 0.05). There were no differences in sensitivity between control and captopril-treated rings with respect to NE, HIST, AI, or AII. Four of the ten captopril-treated segments, however, failed to respond to AI, and the maximal active tension of the responders was significantly reduced (control 0.47 +/- 0.06 g vs. 0.20 +/- 0.05 g; p less than 0.02). These results suggest that ACE is involved in the modulation of vein graft intimal hyperplasia, and that ACE inhibitors may have therapeutic applications in patients undergoing vein bypass procedures.


Morphological and behavioral markers of environmentally induced retardation of brain development: an animal model.
In most neurotoxicological studies morphological assessment focuses on pathological effects, like degenerative changes in neuronal perikarya, axonopathy, demyelination, and glial and endothelial cell reactions. Similarly, the assessment of physiological and behavioral effects center on evident neurological symptoms, like EEG and EMG abnormalities, resting and intention tremor, abnormal gait, and abnormal reflexes. This paper reviews briefly another central nervous system target of harmful environmental agents, which results in behavioral abnormalities without any qualitatively evident neuropathology. This is called microneuronal hypoplasia, a retardation of brain development characterized by a quantitative reduction in the normal population of late-generated, short-axoned neurons in specific brain regions. Correlated descriptive and experimental neurogenetic studies in the rat have established that all the cerebellar granule cells and a very high proportion of hippocampal granule cells are produced postnatally, and that focal, low-dose X-irradiation either of the cerebellum or of the hippocampus after birth selectively interferes with the acquisition of the full complement of granule cells (microneuronal hypoplasia). Subsequent behavioral investigations showed that cerebellar microneuronal hypoplasia results in profound hyperactivity without motor abnormalities, while hippocampal microneuronal hypoplasia results in hyperactivity, as well as attentional and learning deficits. There is much indirect clinical evidence that various harmful environmental agents affecting the pregnant mother and/or the infant lead to such childhood disorders as hyperactivity and attentional and learning disorders. As the developing human brain is more mature at birth than the rat brain, the risk for microneuronal hypoplasia and consequent behavioral disorders may be highest at late stages of fetal development, in prematurely born and small-for-weight infants, and during the early stages of infant development. Recent technological advances in brain imaging techniques make it possible to test this hypothesis and to assess the possible relationship between the degree of retarded brain development and ensuing behavioral disorders.


Antibody response to homologous epitopes of Epstein-Barr virus, Mycobacterium avium subsp. paratuberculosis and IRF5 in patients with different connective tissue diseases and in mouse model of antigen-induced arthritis.
Improved knowledge of different biomarkers is crucial for early diagnosis of rheumatic diseases and to provide important insights for clinical management. In this study, we evaluated the seroreactivity of patients with different connective tissue diseases (CTDs) (rheumatoid arthritis, RA; systemic lupus erythematosus, SLE; systemic sclerosis, SSc; and Sjogren's syndrome, SSj) to interferon regulatory factor 5 (IRF5) peptide and homologs derived from Epstein-Barr virus (EBV) and Mycobacterium avium subsp. paratuberculosis (MAP). Antigen-induced arthritis (AIA) experiments have been performed in control and IRF5 conditional knockout mice to reinforce the hypothesis that antibodies generated against the three homologous peptides are cross-reactive. Reactivity against wild-type (wt) and citrullinated (cit) IRF5 (IRF5424-434), MAP (MAP_402718-32) and EBV (BOLF1305-320) peptides were tested by indirect ELISA in sera from 100 RA patients, 54 patients with other CTDs (14 SLE, 28 SSc and 12 SSj) and 100 healthy subjects (HCs). Antibody responses to the same wt peptides have been tested in AIA mouse sera after immunization with complete Freud's adjuvant (CFA) and methylated bovine serum albumin (mBSA) to induce arthritis in the knee joint. BOLF1, MAP_4027 and IRF5 peptides triggered different antibody responses in CTD diseases with a stronger reactivity in RA (p=0.0001). Similar trends were observed in AIA mice with significantly higher reactivity after 7 days from induction of arthritis. We also found statistically significant differences in antibody responses between SSc and HCs for BOLF1 (p=0.003), MAP_4027 (p=0.0076) and IRF5 (p=0.0042). Peripheral reactivity to cit peptides was lower compared to their wt counterparts, except for cit-MAP_402718-32, which induced stronger responses in RA than wt-MAP_402718-32 (46% vs. 26%, p=0.0170). Conclusion(s): Our results show differential antibody responses to BOLF1, MAP_4027 and IRF5 peptides among CTDs, highlighting their potential as diagnostic biomarkers in these diseases. Experiments performed in IRF5 conditional knockout mice support the hypothesis of cross-reactivity between the investigated homologous antigens.


Papillary lesions of the breast with and without atypical ductal hyperplasia: can we accurately predict benign behavior from core needle biopsy?
Evaluation of papillary lesions of the breast can be difficult, and in core needle biopsy specimens, accurate diagnosis is challenging. Initial studies suggested that all papillary lesions revealed by core biopsy required surgical excision. Recent data suggest that only papillary lesions with atypical ductal hyperplasia (ADH) revealed by core biopsy need surgical excision. We evaluated our experience at the University of Washington Medical Center, Seattle, with papillary lesions with and without ADH on core biopsy to determine whether diagnostic accuracy can be achieved. In 51 core biopsy specimens, we evaluated the presence or absence of ADH: 25 were benign papillomas; 26 were papillomas with ADH. Surgical follow-up was available for 36 cases (11 papillomas and 25 papillomas with ADH). Clinical (radiologic) follow-up was available in 5 papilloma cases (average follow-up, 35.6 months). Follow-up revealed that all papillomas on core biopsy were benign. Excisional biopsy revealed ductal carcinoma in situ or invasive carcinoma in 12 (48%) of 25 papillary lesions with ADH. Benign papillomas can be adequately diagnosed with core biopsy. All papillary lesions with ADH require surgical excision owing to the high rate of associated neoplasia.


The Phylogeny of Sleep Database: A New Resource for Sleep Scientists.
We have constructed a database that describes the sleeping characteristics of 127 different mammalian species representing 46 families across 17 orders. The data were extracted from 178 separate references that were found using standardized search protocols, and for each study includes information on the time spent in REM and NREM sleep, sleep cycle length, the number of animals sampled, their sex and age, and reference citation. Importantly, we also coded nine laboratory condition scores as a way to control for the procedures that were used to collect the data. We created a website that contains the database (http://www.bu.edu/phylogeny/) from which others can both download the data and submit new results. This database is being used to address fundamental questions concerning the evolution of mammalian sleep; similar databases on other groups of organisms will enable sleep biologists to understand patterns of sleep at broader phylogenetic scales.


Lack of a direct effect of estrogen on proliferation and differentiation of normal human osteoblast-like cells.
Although osteoblasts contain estrogen receptors, it is unclear whether estrogen has direct effects on osteoblast proliferation and differentiation. We evaluated the effects of 17 beta-estradiol treatment (1 pM to 10 nM) on the proliferation and differentiation of cultured normal adult human cells that expressed many of the phenotypic characteristics and hormonal sensitivities of mature osteoblasts (hOB cells). Treatment of hOB cells with estradiol for as long as 144 h did not affect the rate of DNA synthesis and had minimal, if any, effects on differentiated function. Whereas alkaline phosphatase activity was increased by nearly twofold (P less than 0.01) when the hOB cells were treated with 1 nM 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3], treatment with estradiol had no effect when given alone and did not affect the cells' response to 1,25-(OH)2D3. Similarly, the release of bone gla protein (BGP, osteocalcin) was induced by treatment with 1,25-(OH)2D3 (P less than 0.05), but estradiol treatment did not affect this response. Cellular levels of mRNA for alkaline phosphatase and BGP were not altered by estradiol treatment. We conclude that estradiol treatment does not have major effects on the growth or differentiation of cultured hOB cells. These results are consistent with previous observations in vivo that indicate estrogen acts principally to decrease bone resorption, not to modulate its formation.


The effect of patient-practitioner communication on pain: a systematic review.
Communication between patients and health care practitioners is expected to benefit health outcomes. The objective of this review was to assess the effects of experimentally varied communication on clinical patients' pain. We searched in July 2012, 11 databases supplemented with forward and backward searches for (quasi-) randomized controlled trials in which face-to-face communication was manipulated. We updated in June 2015 using the four most relevant databases (CINAHL, Cochrane Central, Psychinfo, PubMed). Fifty-one studies covering 5079 patients were included. The interventions were separated into three categories: cognitive care, emotional care, procedural preparation. In all but five studies the outcome concerned acute pain. We found that, in general, communication has a small effect on (acute) pain. The 19 cognitive care studies showed that a positive suggestion may reduce pain, whereas a negative suggestion may increase pain, but effects are small. The 14 emotional care studies showed no evidence of a direct effect on pain, although four studies showed a tendency for emotional care lowering patients' pain. Some of the 23 procedural preparation interventions showed a weak to moderate effect on lowering pain. Different types of communication have a significant but small effect on (acute) pain. Positive suggestions and informational preparation seem to lower patients' pain. Communication interventions show a large variety in quality, complexity and methodological rigour; they often used multiple components and it remains unclear what the effective elements of communication are. Future research is warranted to identify the effective components.


The etched cast restoration--clinical techniques and long-term results.
The etched cast restoration was introduced as the "Maryland bridge" in 1980. The technique has continued to evolve with modifications in tooth structure and framework design as well as improvements in luting resins, including some that chemically bond to the alloy. The conservation of tooth structure and the avoidance of involving the gingival crevice remain the major advantages of the technique. Research has shown that the etched cast restoration, when designed with a retentive framework, exhibits bond strengths comparable with those of traditional complete coverage restorations. The main purpose for this retentive framework is to limit the stresses placed on the resin, thus preventing the phenomenon of stress fatigue in the resin. Clinical observations and an 11-year clinical study involving 127 restorations confirm this research. The long-term success rate in this study was 92.9%.


Revealing the high-resolution three-dimensional network of chromatin and interchromatin space: a novel electron-microscopic approach to reconstructing nuclear architecture.
The nuclear architecture is considered an important contributor to genome function. Although the fine structural features of the cell nucleus have been investigated extensively by means of ultrastructural cytochemistry, mainly on ultrathin sections in two dimensions (2D), there was a of lack routine methods for a rapid reconstruction of three-dimensional (3D) distribution of different structural constituents throughout the nuclear volume. We have now filled this gap by the application of a novel approach associating a pre-embedding selective visualization of nuclear components with a method making use of ultramicrotomy combined with scanning electron microscopy (microtome serial block face scanning electron microscopy--'3View'). We have been able to apply this method to the study of DNA distribution within the nuclear volume and reconstruction of 3D chromatin arrangement in nuclei of rat hepatocytes and endothelial cells. Our observations demonstrate that while chromatin appears to occupy the interior of nuclei rather sparsely on 2D images, once reconstructed in 3D from a series of sequential 2D images it gives the impression of considerably filling the nuclear volume. However, quantitative evaluation of the nuclear volume occupied by DNA in the above two types of nuclei leaves a significant part to the interchromatin space (66.2% for hepatic cells and 41.7% for endothelial cells, including nuclear space occupied by nucleoli). Detailed analysis of the reconstructed nuclei reveals a high degree of superposition of chromatin domains, giving rise to a false impression that they fill a much larger part of the nuclear volume than they really do. Our results show the importance of the contribution of such reconstruction techniques to our understanding of the nuclear architecture.


Urine marking and maternal aggression of wild female mice in relation to anogenital distance at birth.
A series of experiments were conducted with wild house mice to verify the effect of intrauterine position on females' anogenital distance at birth (AGD) and to examine the relationships between a female's AGD, used as a bioassay of androgen exposure during fetal life, and her social behavior and reproductive success in adulthood. Experiment 1 showed that cesarean-delivered females that developed in utero between two males (2 M females) have significantly longer AGD's than females positioned between two females (0 M females). We then categorized naturally delivered females shortly after birth as having a long, medium or short AGD. In adulthood, these females were tested for their behavior towards unfamiliar pups, their rate of urine-marking in response to a variety of social stimuli, postpartum aggression and success in protecting their litters in response to male and female intruders. Adult females with different AGD's at birth did not differ either in their behavior toward pups or in their rate of urine marking. Conversely, males housed across a wire mesh partition from a long-AGD female deposited a higher number of urine marks than those exposed to a short-AGD female. When tested after delivering a litter, long-AGD females displayed more tail-rattling (a component of agonistic behavior) towards intruders of both sexes in comparison to short-AGD females. These results are consistent with the hypothesis that females with a long AGD are exposed to higher levels of Testosterone during fetal life than females with a short AGD. Although not related to AGD, other measures of maternal aggression were affected by postpartum day, sex of intruders and a female's infanticidal potential while a virgin.


Ailanthone inhibits non-small cell lung cancer cell growth through repressing DNA replication via downregulating RPA1.
The identification of bioactive compounds from Chinese medicine plays a crucial role in the development of novel reagents against non-small lung cancer (NSCLC). High throughput screening assay and analyses of cell growth, cell cycle, apoptosis, cDNA microarray, BrdU incorporation and gene expression were performed. Ailanthone (Aila) suppressed NSCLC cell growth and colony formation in vitro and inhibited NSCLC tumour growth in subcutaneously xenografted and orthotopic lung tumour models, leading to prolonged survival of tumour-bearing mice. Moreover, Aila induced cell cycle arrest in a dose-independent manner but did not induce apoptosis in all NSCLC cells. Furthermore, 1222 genes were differentially expressed upon Aila administration, which were involved in 21 signal pathways, such as DNA replication. In addition, Aila dose-dependently decreased BrdU incorporation and downregulated the expression of replication protein A1 (RPA1). Aila inhibited the growth of NSCLC cells through the repression of DNA replication via downregulating RPA1, rather than through cell cycle arrest and apoptosis. Our findings suggested that Aila could be used as a promising therapeutic candidate for NSCLC patients.


High-order perturbation theory for light scattering from a rough metal surface.
The angular distribution of the mean diffuse intensity scattered from a metal surface with one-dimensional roughness is studied with perturbation theory. From an approach based on the reduced Rayleigh equations in p polarization, exact perturbation terms up to eighth order in the height parameter are developed for surface roughness consistent with a stationary Gaussian process. The theory is evaluated for a number of cases in which surface plasmon polariton excitation is significant and produces effects such as backscattering enhancement. For surface roughness having a wide Gaussian power spectrum, it is found that the high-order terms lead to roughness-induced broadening of the backscattering peak. For rectangular spectra, two cases are studied in which backscattering effects are due to sixth- and eighth-order terms; both cases provide good comparisons with previously unexplained experimental results. Further, because of an eighth-order term, the diffuse intensity is shown to contain a specular peak that also relies on polariton excitation. This new effect is studied in detail and is found to arise from the constructive interference of contributions produced by multiple-scattering processes, although the time-reversed paths that produce backscattering enhancement are not essential to the specular effect.


On total disc replacement.
Low back pain consumes a large part of the community's resources dedicated to health care and sick leave. Back disorders also negatively affect the individual leading to pain suffering, decreased quality-of-life and disability. Chronic low back pain (CLBP) due to degenerative disc disease (DDD) is today often treated with fusion when conservative treatment has failed and symptoms are severe. This treatment is as successful as arthroplasty is for hip arthritis in restoring the patient's quality of life and reducing disability. Even so, there are some problems with this treatment, one of these being recurrent CLBP from an adjacent segment (ASD) after primarily successful surgery. This has led to the development of alternative surgical treatments and devices that maintain or restore mobility, in order to reduce the risk for ASD. Of these new devices, the most frequently used are the disc prostheses used in Total Disc Replacement (TDR). This thesis is based on four studies comparing total disc replacement with posterior fusion. The studies are all based on a material of 152 patients with DDD in one or two segments, aged 20-55 years that were randomly treated with either posterior fusion or TDR. The first study concerned clinical outcome and complications. Follow-up was 100% at both one and two years. It revealed that both treatment groups had a clear benefit from treatment and that patients with TDR were better in almost all outcome scores at one-year follow-up. Fusion patients continued to improve during the second year. At two-year follow-up there was a remaining difference in favour of TDR for back pain. 73% in the TDR group and 63% in the fusion group were much better or totally pain-free (n.s.), while twice as many patients in the TDR group were totally pain free (30%) compared to the fusion group (15%). Time of surgery and total time in hospital were shorter in the TDR group. There was no difference in complications and reoperations, except that seventeen of the patients in the fusion group were re-operated for removal of their implants. The second study concerned sex life and sexual function. TDR is performed via an anterior approach, an approach that has been used for a long time for various procedures on the lumbar spine. A frequent complication reported in males when this approach is used is persistent retrograde ejaculation. The TDR group in this material was operated via an extra-peritoneal approach to the retroperitoneal space, and there were no cases of persistent retrograde ejaculation. There was a surprisingly high frequency of men in the fusion group reporting deterioration in ability to have an orgasm postoperatively. Preoperative sex life was severely hampered in the majority of patients in the entire material, but sex life underwent a marked improvement in both treatment groups by the two-year follow-up that correlated with reduction in back pain. The third study was on mobility in the lumbar spinal segments, where X-rays were taken in full extension and flexion prior to surgery and at two-year follow-up. Analysis of the films showed that 78% of the patients in the fusion group reached the surgical goal (non-mobility) and that 89% of the TDR patients maintained mobility. Preoperative disc height was lower than in a normative database in both groups, and remained lower in the fusion group, while it became higher in the TDR group. Mobility in the operated segment increased in the TDR group postoperatively. Mobility at the rest of the lumbar spine increased in both treatment groups. Mobility in adjacent segments was within the norm postoperatively, but slightly larger in the fusion group. In the fourth study the health economics of TDR vs Fusion was analysed. The hospital costs for the procedure were higher for patients in the fusion group compared to the TDR group, and the TDR patients were on sick-leave two months less. In all, these studies showed that the results in the TDR group were as good as in the fusion group. Patients are more likely to be totally pain-free when treated with TDR compared to fusion. Treatment with this new procedure seems justified in selected patients at least in the short-term perspective. Long-term follow-up is underway and results will be published in due course.


Effects of a comprehensive nutrition education programme to change grade 4 primary-school students' eating behaviours in China.
As part of a national initiative to reduce child obesity, a comprehensive school-based nutrition education intervention to change eating behaviours among grade 4 primary-school students was developed, implemented and evaluated. The intervention was developed by school staff, with technical assistance from outside health education specialists. The programme included school facility upgrades, school teacher/staff training, curriculum changes and activities for parents. Student scores on nine key eating behaviours were assessed prior to and after the programme. The quality of programme implementation in the schools was monitored by technical assistance teams. Shandong Province (high household income) and Qinghai Province (low household income), China. Three programme schools and three control schools in each province.ParticipantsStudents in grade 4 (age 8-9 years). There were significant positive changes in self-reported eating behaviour scores from pre- to post-assessment in programme schools. At post-test students in programme schools had significantly higher scores than students in control schools after controlling for other variables. The programme was more effective in the high-income province. Observations by the technical assistance teams suggested the programme was implemented more completely in Shandong. The teams noted the challenges for implementing and evaluating programmes like these. This intervention increased healthy eating behaviours among 4th graders in both provinces and had more effect in the more affluent province. Results suggest that a scaled-up initiative using existing school and public health resources could change eating practices in a large population over time. The intervention also provided lessons for implementing and evaluating similar nutrition programmes.