[Changes in the use of hospital resources from prison: a 16 year study].
To describe hospital morbidity caused by the inmates of our prison over the past 16 years. retrospective study of hospital admissions between 01-01-1994 and 31-12-2009, divided into four periods. Socio-demographic variables were collected: duration of stay and discharge diagnosis. Quantitative variables were described as means and medians and qualitative variables as absolute and relative frequencies. A mean comparison was performed on quantitative variables and qualitative proportions. For equal variances, an ANOVA test was performed with linear trend study of encoding the variable "period" with orthogonal contrasts. Without equality of variances, comparisons were made using the Kruskal-Wallis test, and tendencies by means of the nonparametric Jonckheere-Terpstra test. For qualitative variables we used the Pearson Chi-Square, evaluating the trend with the chi-square for linear trend. 625 patients generated 996 admissions with no temporal variation. The median age is 33 years, with an upward trend (29 years to 38, p <0.0001). 47.9% were HIV + [(63.3% to 35.9%), p <0.0001]. The average stay was 9.6 days (95% CI 8.8 to 10.4) [11.9 (10.0 to 13.9) 9.6 (8.8 to 10.4), p = 0.002]. The frequency of internal and year 1000 remained unchanged (111.6 to 87.9, p = 0.366). The days of hospitalization decreased (3066 to 2442, p = 0.049)) and the average admitted per day (2.1 to 1.7, p = 0.049). The use of hospital resources from prison is constant. The way they use it has changed along with the pathology that causes it. HIV is no longer the primary pathology.


Development and implementation of South Asia's first heat-health action plan in Ahmedabad (Gujarat, India).
Recurrent heat waves, already a concern in rapidly growing and urbanizing South Asia, will very likely worsen in a warming world. Coordinated adaptation efforts can reduce heat's adverse health impacts, however. To address this concern in Ahmedabad (Gujarat, India), a coalition has been formed to develop an evidence-based heat preparedness plan and early warning system. This paper describes the group and initial steps in the plan's development and implementation. Evidence accumulation included extensive literature review, analysis of local temperature and mortality data, surveys with heat-vulnerable populations, focus groups with health care professionals, and expert consultation. The findings and recommendations were encapsulated in policy briefs for key government agencies, health care professionals, outdoor workers, and slum communities, and synthesized in the heat preparedness plan. A 7-day probabilistic weather forecast was also developed and is used to trigger the plan in advance of dangerous heat waves. The pilot plan was implemented in 2013, and public outreach was done through training workshops, hoardings/billboards, pamphlets, and print advertisements. Evaluation activities and continuous improvement efforts are ongoing, along with plans to explore the program's scalability to other Indian cities, as Ahmedabad is the first South Asian city to address heat-health threats comprehensively.


Reduction of MDSCs with All-trans Retinoic Acid Improves CAR Therapy Efficacy for Sarcomas.
Genetically engineered T cells expressing CD19-specific chimeric antigen receptors (CAR) have shown impressive activity against B-cell malignancies, and preliminary results suggest that T cells expressing a first-generation disialoganglioside (GD2)-specific CAR can also provide clinical benefit in patients with neuroblastoma. We sought to assess the potential of GD2-CAR therapies to treat pediatric sarcomas. We observed that 18 of 18 (100%) of osteosarcomas, 2 of 15 (13%) of rhabdomyosarcomas, and 7 of 35 (20%) of Ewing sarcomas expressed GD2. T cells engineered to express a third-generation GD2-CAR incorporating the 14g2a-scFv with the CD28, OX40, and CD3ζ signaling domains (14g2a.CD28.OX40.ζ) mediated efficient and comparable lysis of both GD2+ sarcoma and neuroblastoma cell lines in vitro However, in xenograft models, GD2-CAR T cells had no antitumor effect against GD2+ sarcoma, despite effectively controlling GD2+ neuroblastoma. We observed that pediatric sarcoma xenografts, but not neuroblastoma xenografts, induced large populations of monocytic and granulocytic murine myeloid-derived suppressor cells (MDSC) that inhibited human CAR T-cell responses in vitro Treatment of sarcoma-bearing mice with all-trans retinoic acid (ATRA) largely eradicated monocytic MDSCs and diminished the suppressive capacity of granulocytic MDSCs. Combined therapy using GD2-CAR T cells plus ATRA significantly improved antitumor efficacy against sarcoma xenografts. We conclude that retinoids provide a clinically accessible class of agents capable of diminishing the suppressive effects of MDSCs, and that co-administration of retinoids may enhance the efficacy of CAR therapies targeting solid tumors. Cancer Immunol Res; 4(10); 869-80. ©2016 AACR.


Determinants of Macular Layers and Optic Disc Characteristics on SD-OCT: The Rhineland Study.
To investigate variation and determinants of macular layers, peripapillary retinal nerve fiber layer (pRNFL) and Bruch's membrane opening-minimum rim width (BMO-MRW) in the general population. In 1306 participants, we performed spectral domain optical coherence tomography (SD-OCT) scans of the macula, pRNFL, and BMO-MRW, and assessed their determinants using multivariable regression. Intraindividual interocular differences were analyzed using Spearman's rank correlation analysis. Participant age ranged from 30 to 95 years (mean ± standard deviation, 56.1 ± 13.9) and 56% were women. Interocular correlation ranged from 0.63 to 0.93. Differences increased with age and were larger in persons with glaucoma or prior stroke. pRNFL and BMO-MRW decreased with increasing age. Except for RNFL, volumes of various inner macular layers and the outer nuclear layer (ONL) decreased with increasing age, more negative spherical equivalent (SE), and were lower in women compared to men. For some layers, age effects amplified over the life course. History of stroke was associated with smaller volumes of various layers, without reaching statistical significance. We found no association of further systemic parameters with any SD-OCT parameter. We provide large-scale normative data from a Caucasian general population for various SD-OCT measures. Interocular variability increased with age and specific pathology. Factors, such as age, sex, refraction, and a history of stroke, were associated with various retinal assessments. In clinical routine, our findings should be considered on a per eye basis when interpreting SD-OCT volumes, pRNFL, or BMO-MRW to avoid confounded results.


Use of a learning network to improve variation in interstage weight gain after the Norwood operation.
Growth failure is common in infants with single ventricle. This study evaluated the use of a learning network, the National Pediatric Cardiology Quality Improvement Collaborative (NPC-QIC), to spread optimized nutritional practices and improve infant growth. A previously identified Nutritional Bundle was spread among NPC-QIC sites. interstage weight-for-age z-score change (ΔWAZ) between discharge from stage 1 palliation (S1) and stage 2 surgical palliation (S2). Variation among sites in interstage ΔWAZ was evaluated before (Period 1) and after (Period 2) spread of Nutritional Bundle. We performed an analysis of NPC-QIC registry infants presenting for S2 at sites previously shown to have significant variation in interstage patient growth. Four hundred seven infants from 15 sites underwent S2 between 2008 and 2013: 158 in Period 1 (December 2008-December 2010) and 249 in Period 2 (December 2010-April 2013). Median age at S2 was 4.9 months (2.6-12.8) with no difference between periods. There was significant variation in interstage ΔWAZ among sites in Period 1 (P = .01) but not in Period 2 (P = .39). More patients had an interstage ΔWAZ <0 in Period 1 (43%) than Period 2 (32%) (P = .03). In Period 1, the median interstage ΔWAZ was <0 in six sites while in Period 2 no site had median interstage ΔWAZ <0. Sites with the worst patient growth in Period 1 had marked improvement in Period 2 (P = .02, .06, and .06, respectively). Spread of optimal nutritional practices led to decreased variation in interstage growth with most improvement observed at sites with the worst baseline growth outcomes.


Litebamine, a phenanthrene alkaloid from the wood of Litsea cubeba, inhibits rat smooth muscle cell adhesion and migration on collagen.
Smooth muscle cells (SMCs) play an important role in the development of atherosclerosis and restenosis after angioplasty and coronary bypass grafting. The pathogenesis of these vascular diseases includes the abnormal production of extracellular matrix (ECM) proteins by SMCs and their interactions with this newly synthesized and preexisting ECM. Litebamine, a natural phenanthrene alkaloid from the wood of Litsea cubeba, has been shown to inhibit platelet aggregation and thromboxane B(2) formation, suggesting its antithrombotic activity. In the present study we examined litebamine effects on vascular SMC adhesion and migration. Our results indicated that litebamine inhibited rat aortic SMCs (RASMCs) and A10 thoracic SMCs adhesion to collagen but not to other matrix proteins, suggesting its specificity on collagen. This inhibition was possibly resulted from that litebamine attenuated immobilized collagen-induced focal adhesion kinase (FAK) phosphorylation and actin cytoskeleton reorganization in RASMCs, as determined by Western blotting and immunofluorescence microscopy. In a functional study, litebamine also inhibited platelet-derived growth factor (PDGF)-induced RASMC migration but did not affect PDGF-induced matrix metalloproteinases (MMPs) secretion. Strikingly, among the tested kinases involved in PDGF-induced migration, only PDGF-induced phosphatidylinositol-3 kinase (PI-3K) activation was inhibited by litebamine. Taken together, we demonstrated here that litebamine can functionally inhibit vascular SMC adhesion and migration and elucidated its possible mechanisms of action. As SMC adhesion and migration are critical events in disease-related vascular remodeling, this compound may have beneficial effects in preventing cardiovascular diseases.


Paying the price in an era of HIV treatment as prevention: a retrospective study of the cost burden of HIV treatment for people living with HIV in Victoria, Australia.
Background An estimated 25700 people live with diagnosed HIV (PLWH) in Australia and ~1200 newly diagnosed cases were notified in 2012. New HIV prevention strategies focus on individual uptake of treatment; however, a potential barrier is the financial burden of antiretroviral treatment (ART). We describe HIV ART dispensed and the estimated associated costs for PLWH in Victoria. A retrospective cross-sectional study of pharmacy data on ART dispensed between January 2012 and November 2013 from a hospital network, including Victoria's largest sexual health clinic was conducted. Estimated annual patient costs of ART were calculated by the number of items dispensed per year, concession status, dispensing site and applicable co-payment. A total of 60225 dispensing records from 3903 individuals were included; this represented 83.8% of pharmaceutical benefits scheme-recorded ART dispensed in Victoria over this period. The estimated annual co-payment costs for patients without a concession card and who were collecting two medications was $433.20. One-fifth of patients (21.3%) collected four or more items, equating to an estimated annual cost of at least $866.40 without a concession card and $141.60 with a concession card. Of those dispensed four or more items, 40.4% were concession card holders. There may be meaningful patient costs associated with accessing ART for some PLWH. New HIV treatment-based prevention strategies need to consider financial vulnerabilities and appropriately targeted initiatives to alleviate patient costs associated with ART, ensuring they do not act as a barrier to commencement of and adherence to HIV treatment.


Lacunar infarcts: no black holes in the brain are benign.
Lacunar infarcts--small subcortical infarcts that result from occlusion of a single penetrating artery--account for about one quarter of all ischaemic strokes. However, there are many diagnostic pitfalls, and causes other than penetrating small vessel disease in up to one third of cases. Recent studies have shown that the prognosis after lacunar infarcts is not benign; the risk of recurrent stroke is no lower than for other ischaemic stroke subtypes, and there is an increased risk for cognitive decline, dementia and death in the long term. Furthermore, silent small vessel disease in the brain at the time of an index stroke has significant prognostic implications. In the acute phase, response to intravenous thrombolysis appears to be similar to other subtypes of ischaemic strokes. Antiplatelet drugs, careful blood pressure control, statins and modification of lifestyle risk factors are key elements in secondary prevention after lacunar infarcts.


The Prospective Oral Mucositis Audit: relationship of severe oral mucositis with clinical and medical resource use outcomes in patients receiving high-dose melphalan or BEAM-conditioning chemotherapy and autologous SCT.
The Prospective Oral Mucositis Audit was an observational study in 197 patients with multiple myeloma (MM) or non-Hodgkin's lymphoma (NHL) undergoing, respectively, high-dose melphalan or BEAM chemotherapy and autologous SCT at 25 European centres. We evaluated the relationship between severe oral mucositis (SOM; WHO Oral Toxicity Scale grade 3-4) and local and systemic clinical sequelae and medical resource use. SOM occurred in 44% of patients. The duration of SOM (mean 5.3 days) correlated with time to neutrophil engraftment. The following parameters increased gradiently with maximum grade of oral mucositis: duration of pain score >or=4, opioid use, dysphagia score >or=4, total parenteral nutrition (TPN) use, incidence and/or duration of fever and infection, and duration of antibiotic use. SOM increased the duration of TPN use by 2.7 days (P<0.001), opioids by 4.6 days (P<0.001), and antibiotics by 2.4 days (P=0.045). SOM prolonged hospital stay by 2.3 days (P=0.013) in MM patients, but not in NHL patients (who tended to have a longer hospital stay). In conclusion, this analysis of prospectively collected observational data provides important insight into the scope and impact of SOM in the European transplant setting.


Emergent Damped Oscillation Induced by Nutrient-Modulating Growth Feedback.
Growth feedback, the inherent coupling between the synthetic gene circuit and the host cell growth, could significantly change the circuit behaviors. Previously, a diverse array of emergent behaviors, such as growth bistability, enhanced ultrasensitivity, and topology-dependent memory loss, were reported to be induced by growth feedback. However, the influence of the growth feedback on the circuit functions remains underexplored. Here, we reported an unexpected damped oscillatory behavior of a self-activation gene circuit induced by nutrient-modulating growth feedback. Specifically, after dilution of the activated self-activation switch into the fresh medium with moderate nutrients, its gene expression first decreases as the cell grows and then shows a significant overshoot before it reaches the steady state, leading to damped oscillation dynamics. Fitting the data with a coarse-grained model suggests a nonmonotonic growth-rate regulation on gene production rate. The underlying mechanism of the oscillation was demonstrated by a molecular mathematical model, which includes the ribosome allocation toward gene production, cell growth, and cell maintenance. Interestingly, the model predicted a counterintuitive dependence of oscillation amplitude on the nutrition level, where the highest peak was found in the medium with moderate nutrients, but was not observed in rich nutrients. We experimentally verified this prediction by tuning the nutrient level in the culture medium. We did not observe significant oscillatory behavior for the toggle switch, suggesting that the emergence of damped oscillatory behavior depends on circuit network topology. Our results demonstrated a new nonlinear emergent behavior mediated by growth feedback, which depends on the ribosome allocation between gene circuit and cell growth.


Promoting healthy eating and physical activity among school children: findings from Health-E-PALS, the first pilot intervention from Lebanon.
In Lebanon, childhood obesity doubled during the past decade. Preventive measures should start early in life and Schools are considered an important environment to promote energy balance health behaviours. School-based programmes promoting healthy lifestyles are lacking. The purpose of this study was to evaluate the feasibility and effectiveness of a multicomponent school-based intervention to promote healthy eating and physical activity (and prevent obesity) with school children aged 9-11 years in Lebanon. The intervention was developed based on the constructs of the Social Cognitive Theory and adapted to the culture of Lebanese and Arab populations. It consisted of three components: class curriculum, family involvement and food service. Eight schools were purposively selected from two communities of different socioeconomic status (SES) in Beirut and, within each school type, were matched on SES, religious sect profile, and then randomly assigned to either the intervention or control group. Anthropometric measurements and questionnaires on determinants of behavioural change, eating and physical activity habits were completed by the students in both groups at baseline and post intervention. Focus group interviews were conducted in intervention schools at the end of the study. Challenges encountered during the programme implementation were also identified, since Lebanon is considered a country with political unrest and no similar research projects were conducted in the area. Students in the intervention group reported purchasing and consuming less chips and sweetened drinks post-intervention compared with controls (86% & 88% less respectively p < 0.001). Knowledge and self-efficacy scores increased for the intervention (+2.8 & +1.7 points respectively p < 0.001) but not for the control group. There was no difference in physical activity and screen time habits and no changes in BMI between groups at post intervention. Interview data from focus groups showed that the programme was generally well accepted. Limitations for better outcomes include the length of the programme and the school environment. "Health-E-PALS" intervention is a promising innovative, theory-based, culturally sensitive intervention to promote healthy eating habits and physical activity in Lebanese school children with a potential to be scaled up, replicated and sustained.


Novel pancreatic cancer cell lines derived from genetically engineered mouse models of spontaneous pancreatic adenocarcinoma: applications in diagnosis and therapy.
Pancreatic cancer (PC) remains one of the most lethal human malignancies with poor prognosis. Despite all advances in preclinical research, there have not been significant translation of novel therapies into the clinics. The development of genetically engineered mouse (GEM) models that produce spontaneous pancreatic adenocarcinoma (PDAC) have increased our understanding of the pathogenesis of the disease. Although these PDAC mouse models are ideal for studying potential therapies and specific genetic mutations, there is a need for developing syngeneic cell lines from these models. In this study, we describe the successful establishment and characterization of three cell lines derived from two (PDAC) mouse models. The cell line UN-KC-6141 was derived from a pancreatic tumor of a Kras(G12D);Pdx1-Cre (KC) mouse at 50 weeks of age, whereas UN-KPC-960 and UN-KPC-961 cell lines were derived from pancreatic tumors of Kras(G12D);Trp53(R172H);Pdx1-Cre (KPC) mice at 17 weeks of age. The cancer mutations of these parent mice carried over to the daughter cell lines (i.e. Kras(G12D) mutation was observed in all three cell lines while Trp53 mutation was observed only in KPC cell lines). The cell lines showed typical cobblestone epithelial morphology in culture, and unlike the previously established mouse PDAC cell line Panc02, expressed the ductal marker CK19. Furthermore, these cell lines expressed the epithelial-mesenchymal markers E-cadherin and N-cadherin, and also, Muc1 and Muc4 mucins. In addition, these cell lines were resistant to the chemotherapeutic drug Gemcitabine. Their implantation in vivo produced subcutaneous as well as tumors in the pancreas (orthotopic). The genetic mutations in these cell lines mimic the genetic compendium of human PDAC, which make them valuable models with a high potential of translational relevance for examining diagnostic markers and therapeutic drugs.


Lentiviral vectors: turning a deadly foe into a therapeutic agent.
The past 3 years have witnessed the spectacular irruption of lentiviral vectors into the limelight of the gene therapy scene. Owing to their ability to deliver transgenes in tissues that had long appeared irremediably refractory to stable genetic manipulation, lentivectors have opened fresh perspectives for the genetic treatment of a wide array of hereditary as well as acquired disorders, and a concrete proposal for their clinical use seems imminent. This article traces the path that has led to this rapid development and describes the current state of the art in the design and production of lentiviral vectors. The important question of biosafety is discussed. This system seems to have the edge over other gene delivery tools for particular targets, however, there remain several issues to be resolved before lentivectors make it to the bedside. Gene Therapy (2000) 7, 20-23.


Ultrasmall endogenous biopolymer nanoparticles for magnetic resonance/photoacoustic dual-modal imaging-guided photothermal therapy.
Multi-modal imaging-guided photothermal therapy (PTT) has aroused extensive attention in biomedical research recently because it can provide more comprehensive information for accurate diagnosis and treatment. In this research, the manganese ion chelated endogenous biopolymer melanin nanoparticles were successfully prepared for magnetic resonance (MR)/photoacoustic (PA) dual-modal imaging-guided PTT. The obtained nanoparticles with an ultrasmall size of about 3.2 nm exhibited negligible cytotoxicity, high relaxivity for MRI, an excellent photothermal effect and PA activity. Moreover, in vivo MRI and PAI results all demonstrated that the nanoparticles began to diffuse in the blood after intratumoral injection into tumor-bearing mice and could spread throughout the whole tumor region at 3 h, indicating the optimal treatment time. The subsequent photothermal therapy of cancer cells in vivo was carried out and the result showed that tumor growth could be effectively inhibited without inducing any observed side effects. Besides, melanin as an endogenous biopolymer has native biocompatibility and biodegradability, and it can be excreted through both renal and hepatobiliary pathways after treatment. Therefore, the melanin-Mn nanoparticles may assist in better indicating the optimal treatment time, monitoring the therapeutic process and enhancing the therapeutic effect and showed great clinical translation potential for cancer diagnosis and therapy.


Sexual intercourse and oral sex among public middle school students: prevalence and correlates.
Early sexual initiation is associated with elevated teenage pregnancy and STD risk, yet little is known about the prevalence and correlates of sexual behavior among young adolescents. Better information is needed to guide interventions to prevent early sexual debut. Data from a 2005 survey of 4,557 sixth-, seventh- and eighth-grade students at 14 urban public schools in Southern California were analyzed using chi-square tests and logistic regression, to identify correlates of oral sex, intercourse and both. Overall, 9% of youth had ever had sexual intercourse, and 8% had had oral sex. Three percent reported having had oral sex only, 4% intercourse only and 5% both. Among those who reported intercourse, 69% had used a condom at last intercourse, and 43% had had multiple partners. Being male, being black and having at least one friend who had ever been involved in a pregnancy were positively associated with having had intercourse only and both intercourse and oral sex (odds ratios, 1.7-4.2). Being in eighth grade, expecting to have intercourse in the next six months and currently having a boyfriend or girlfriend were positively associated with all three outcomes (2.1-7.2). Intercourse and oral sex were highly correlated. Interventions addressing oral sex, intercourse and multiple partners should begin before sixth grade and continue throughout the middle school years. Health professionals should target adolescent risk reduction counseling toward males, blacks, youth with a boyfriend or girlfriend, and those with a friend who has been involved in a pregnancy.


Transitioning sediment quality assessment into regulations: Challenges and solutions in implementing California's sediment quality objectives.
Development and promulgation of sediment quality criteria represents a substantial challenge for water quality agencies. Unlike water quality programs that rely on individual chemical thresholds to assess water quality, the complex processes affecting contaminant bioavailability in sediments preclude the use of contaminant concentrations to independently assess impacts or identify cause. Various multiple line of evidence approaches (e.g., sediment quality triad) have been developed for sediment quality assessment, but such frameworks are rarely fully incorporated into statewide regulatory programs due to a lack of standardized and validated tools. In 2003, California's State Water Resources Control Board (State Water Board) initiated development of sediment quality criteria and an assessment framework that required the developers to resolve many challenging technical and policy related issues to the satisfaction of stakeholders, scientists, and the general public. The first part of this multiyear effort has been completed and resulted in the development and validation of an integrated collection of tools, thresholds, and a data interpretation framework for assessing sediment contamination impacts on benthic community condition. The State Water Board's narrative sediment quality criteria and assessment framework became effective in 2009, following US Environmental Protection Agency approval. The results of this effort are described in a series of 6 articles published in this issue of Integrated Environmental Assessment and Management. The articles describe: 1) a multiple line of evidence framework for data integration and assessment, 2) calibration and evaluation of sediment quality guidelines for predicting toxic responses, 3) development and evaluation of sediment quality guidelines with respect to benthic macrofauna responses, 4) selection of toxicity test methods and thresholds, 5) identification and characterization of benthic community assemblages, 6) the effect of sampling methods on benthic community assessment, and 7) recommendations on improving the assessment of contaminant exposure in sediment quality assessment. This collection of articles illustrates the steps needed to improve the scientific foundation for sediment quality assessment in regulatory applications.


Influence of Acanthamoeba castellanii on intracellular growth of different Legionella species in human monocytes.
Previous studies using a murine model of coinhalation of Legionella pneumophila and Hartmannella vermiformis have shown a significantly enhanced intrapulmonary growth of L. pneumophila in comparison to inhalation of legionellae alone (J. Brieland, M. McClain, L. Heath, C. Chrisp, G. Huffnagle, M. LeGendre, M. Hurley, J. Fantone, and C. Engleberg, Infect. Immun. 64:2449-2456, 1996). In this study, we introduce an in vitro coculture model of legionellae, Mono Mac 6 cells (MM6) and Acanthamoeba castellanii, using a cell culture chamber system which separates both cell types by a microporous polycarbonate membrane impervious to bacteria, amoebae, and human cells. Whereas L. pneumophila has shown a maximal 4-log-unit multiplication within MM6, which could not be further increased by coculture with Acanthamoeba castellanii, significantly enhanced replication of L. gormanii, L. micdadei, L. steigerwaltii, L. longbeachae, and L. dumoffii was seen after coculture with amoebae. This effect was seen only with uninfected amoebae, not with Legionella-infected amoebae. The supporting effect for intracellular multiplication in MM6 could be reproduced in part by addition of a cell-free coculture supernatant obtained from a coincubation experiment with uninfected A. castellanii and Legionella-infected MM6, suggesting that amoeba-derived effector molecules are involved in this phenomenon. This coculture model allows investigations of molecular and biochemical mechanisms which are responsible for the enhancement of intracellular multiplication of legionellae in monocytic cells after interaction with amoebae.


Domain-specific impairment in metacognitive accuracy following anterior prefrontal lesions.
Humans have the capacity to evaluate the success of cognitive processes, known as metacognition. Convergent evidence supports a role for anterior prefrontal cortex in metacognitive judgements of perceptual processes. However, it is unknown whether metacognition is a global phenomenon, with anterior prefrontal cortex supporting metacognition across domains, or whether it relies on domain-specific neural substrates. To address this question, we measured metacognitive accuracy in patients with lesions to anterior prefrontal cortex (n = 7) in two distinct domains, perception and memory, by assessing the correspondence between objective performance and subjective ratings of performance. Despite performing equivalently to a comparison group with temporal lobe lesions (n = 11) and healthy controls (n = 19), patients with lesions to the anterior prefrontal cortex showed a selective deficit in perceptual metacognitive accuracy (meta-d'/d', 95% confidence interval 0.28-0.64). Crucially, however, the anterior prefrontal cortex lesion group's metacognitive accuracy on an equivalent memory task remained unimpaired (meta-d'/d', 95% confidence interval 0.78-1.29). Metacognitive accuracy in the temporal lobe group was intact in both domains. Our results support a causal role for anterior prefrontal cortex in perceptual metacognition, and indicate that the neural architecture of metacognition, while often considered global and domain-general, comprises domain-specific components that may be differentially affected by neurological insult.


Population genetic structure of the African malaria mosquito Anopheles funestus in Kenya.
Anopheles funestus Giles is a major malaria vector in Africa, but little is known about the genetic structure of natural populations. In this study, microsatellite markers were used to investigate the genetic structure of A. funestus populations from Kenya. Two populations from western Kenya 80 km apart and two from coastal Kenya 50 km apart were collected and examined for allelic variation at five trinucleotide microsatellite loci. We found A. funestus Giles was the predominant species (> 98%) in the A. funestus group in these populations. The western Kenya populations exhibited higher genetic diversity than the coastal populations. No significant genetic structure for populations within the coastal or western Kenya regions was detected. However, population genetic differentiation between the two regions was high (F(ST) = 0.208, R(ST) = 0.158), approximately two-fold higher than A. gambiae populations from the same area. The results suggest that the minimum area associated with a deme of A. funestus in western or coastal Kenya is larger than 50 km in diameter. The Great Rift Valley in east Africa, high-elevation mountains in western Kenya, and the vast arid area east to the Great Rift Valley may all play a role in restricting A. funestus gene flow between coastal and western Kenya.


Epidural anaesthesia and long term backache after childbirth.
To examine the relation between epidural anaesthesia and long term backache after childbirth. Data from postal questionnaire on morbidity after childbirth sent to women who had delivered in one maternity hospital between 1978 and 1985 were linked to maternity case notes for each woman. Maternity hospital in Birmingham. 11,701 Women who had delivered their most recent baby at the maternity hospital during the defined period and who returned their completed questionnaires. Of the 1634 women who reported backache, 1132 (69%) had had it for over a year. A significant association was found between backache and epidural anaesthesia (relative risk = 1.8); 903 of 4766 women (18.9%) who had had epidural anaesthesia reported this symptom, compared with 731 of the 6935 women (10.5%) who had not had epidural anaesthesia. This association was consistent in both "normal" and "abnormal" deliveries, the only exception being after an elective caesarean section when no excess backache occurred after epidural anaesthesia. The relation between backache and epidural anaesthesia is probably causal. It seems to result from a combination of effective analgesia and stressed posture during labour. Further investigations on the mechanisms causing backache after epidural anaesthesia are required.


Assessing for unique immunomodulatory and neuroplastic profiles of physical activity subtypes: a focus on psychiatric disorders.
Physical activity (PA) is emerging as a safe and effective tool in the prevention and treatment of psychiatric disorders. PA subtypes include aerobic, resistance, flexibility, neuromotor (involving balance, agility and co-ordination), mind-body (e.g. tai chi, qi gong and yoga) and mixed type trainings. Evidence from clinical trials suggests that PA subtypes can have positive clinical effects, however the effects on the symptomatology may vary according to the PA subtype. It therefore stands to reason that various PA subtypes may modulate the immune system and neuroplastic processes differently. This systematic review aims to assess the immunomodulatory and neuroplastic profiles of various PA subtypes, particularly in unipolar depression and age-related cognitive decline (ARCD). The literature suggests several unique immunomodulatory and neuroplastic profiles for PA subtypes (i.e. resistance, aerobic and mind-body) in depression and ARCD. In depression, levels of various cytokines at baseline may predict treatment response to subtypes of PA and pharmacological agents. The pro-neuroplastic effects of resistance and aerobic PA in ARCD may differ due to variances in neurotrophin profiles. At this stage of literature in the field, it is difficult to draw firm conclusions on the specific immunomodulatory and neuroplastic pathways involved in these PA subtypes given of the small number of comparative studies and methodological heterogeneity between studies (e.g. study population age and illness severity, as well as duration and intensity of PA intervention). This important field requires well-designed, high-quality comparative studies to better describe unique immunomodulatory and neuroplastic profiles.


[Polygenic risk prediction of common diseases: from epidemiology to clinical application].
Since the first map of the human genome was published in 2001 our knowledge about our genetic code has increased exponentially. In addition to high-risk genes for monogenic diseases, such as Huntington's disease and cystic fibrosis, for a number of common diseases, such as breast cancer and cardiovascular disease, many genetic variants that each have a slight increased-risk effect, have been identified via genome-wide association studies (GWAS). A polygenic risk score (PRS) can be calculated on the basis of these single-nucleotide polymorphisms (SNPs), by which an increasingly accurate prediction can be made of an individual's risk for diseases. The results of epidemiological studies in which a PRS is used to predict an individual's total genetic risk for particular diseases are promising. In the future, the PRS could be a valuable addition to traditional monogenic tests. It is, however, important that the predictive value of a genetic risk profile increases further and that it becomes more clear how a clinician must interpret this type of genetic profile - in combination with traditional risk factors.


The contribution of brown vegetation to vegetation dynamics.
Indices of vegetation dynamics that include both green vegetation (GV) and non-photosynthetic vegetation (NPV), that is, brown vegetation, were applied to MODIS surface reflectance data from 2000 to 2006 for the southwestern United States. These indices reveal that the cover of NPV, a measure of vegetation brownness and a component of ecosystems worldwide, is highly variable in both space and time in the study region. In the more mesic regions of the study area, the timing of peaks in NPV appears to result from simple senescence of GV at the end of the growing season. In these regions, the amplitude of GV cyclicity dominates the total vegetation signal. In contrast, in arid and semiarid regions, the amplitude of cyclicity of NPV dominates the total vegetation signal, showing the vegetation of these regions to be unexpectedly dynamic. Shrublands of southwestern United States exhibit temporal behavior in which the annual peak in NPV cover precedes the annual peak in GV cover by a few months. Several explanations for this behavior are offered. This study shows the importance of vegetation indices that include NPV, or vegetation brownness, in understanding terrestrial ecosystem dynamics, as well as the response to change for these ecosystems.


Demography and life history of the egg parasitoid, Trichogramma brassicae, on two moths Anagasta kuehniella and Plodia interpunctella in the laboratory.
The egg parasitoid, Trichogramma brassicae Bezdenko (Hymenoptera: Trichogrammatidae) is the most important and widely distributed species of Trichogramma in Iran. It attacks eggs of several lepidopterous pests, and is a major biological control agent. Rearing parasitoids is necessary for experimental work, and, potentially, for mass release in the field. Selecting a suitable host is critical for developing a successful rearing method. If other conditions are the same, the rate of population increase will be a suitable indicator of parasitoid performance on different hosts. However, conclusions based on a single generation can be misleading because of the learning ability of parasitoids. Life history parameters of T. brassicae were studied on two hosts easily reared in the laboratory, Anagasta kuehniella Zeller, and Plodia interpunctella Hübner (Lepidoptera: Pyralidae). All the experiments were carried out at 24 +/- 1 degrees C, 65+/-10% RH, and 16:8 L:D photoperiod. Eight parameters including gross and net reproductive rates (GRR and R(0) respectively), intrinsic rate of natural increase (r(m)), finite rate of population increase (lambda), intrinsic birth and death rates (b and d respectively), cohort generation time (T), and doubling time (DT) were compared between two hosts for two generations. All parameters showed a highly significant difference (alpha = 0.01) between hosts. GRR, R(0), r(m), lambda, and b were higher, while d, T, and DT were lower in Anagasta than Plodia. The intrinsic rate of natural increase was 0.2912 and 0.2145 female/female/day and net replacement rate was 45.51 and 19.26 female/female/generation in Anagasta and Plodia respectively. Differences between generations were significant except for r(m), lambda, and d. The net replacement rate was 28.56 and 39 in the 1(st) and 2(nd) generations respectively. These results showed that A. kuehniella was a better host than P. interpunctella. Higher reproduction occurred in the second generation that may be due to increased adaptation to experimental conditions.


Gene expression profiling of histiocytic sarcomas in a canine model: the predisposed flatcoated retriever dog.
The determination of altered expression of genes in specific tumor types and their effect upon cellular processes may create insight in tumorigenesis and help to design better treatments. The Flatcoated retriever is a dog breed with an exceptionally high incidence of histiocytic sarcomas. The breed develops two distinct entities of histiocytic neoplasia, a soft tissue form and a visceral form. Gene expression studies of these tumors have value for comparable human diseases such as histiocytic/dendritic cell sarcoma for which knowledge is difficult to accrue due to their rare occurrence. In addition, such studies may help in the search for genetic aberrations underlying the genetic predisposition in this dog breed. Microarray analysis and pathway analyses were performed on fresh-frozen tissues obtained from Flatcoated retrievers with localized, soft tissue histiocytic sarcomas (STHS) and disseminated, visceral histiocytic sarcomas (VHS) and on normal canine spleens from various breeds. Expression differences of nine genes were validated with quantitative real-time PCR (qPCR) analyses. QPCR analyses identified the significantly altered expression of nine genes; PPBP, SpiC, VCAM1, ENPEP, ITGAD (down-regulated), and GTSF1, Col3a1, CD90 and LUM (up-regulated) in the comparison of both the soft tissue and the visceral form with healthy spleen. DAVID pathway analyses revealed 24 pathways that were significantly involved in the development of HS in general, most of which were involved in the DNA repair and replication process. This study identified altered expression of nine genes not yet implicated in histiocytic sarcoma manifestations in the dog nor in comparable human histiocytic/dendritic sarcomas. Exploration of the downside effect of canine inbreeding strategies for the study of similar sarcomas in humans might also lead to the identification of genes related to these rare malignancies in the human.


Biopsychosocial influence on exercise-induced injury: genetic and psychological combinations are predictive of shoulder pain phenotypes.
Chronic pain is influenced by biological, psychological, social, and cultural factors. The current study investigated potential roles for combinations of genetic and psychological factors in the development and/or maintenance of chronic musculoskeletal pain. An exercise-induced shoulder injury model was used, and a priori selected genetic (ADRB2, COMT, OPRM1, AVPR1 A, GCH1, and KCNS1) and psychological (anxiety, depressive symptoms, pain catastrophizing, fear of pain, and kinesiophobia) factors were included as predictors. Pain phenotypes were shoulder pain intensity (5-day average and peak reported on numerical rating scale), upper extremity disability (5-day average and peak reported on the QuickDASH), and shoulder pain duration (in days). After controlling for age, sex, and race, the genetic and psychological predictors were entered as main effects and interaction terms in separate regression models for the different pain phenotypes. Results from the recruited cohort (N = 190) indicated strong statistical evidence for interactions between the COMT diplotype and 1) pain catastrophizing for 5-day average upper extremity disability and 2) depressive symptoms for pain duration. There was moderate statistical evidence for interactions for other shoulder pain phenotypes between additional genes (ADRB2, AVPR1 A, and KCNS1) and depressive symptoms, pain catastrophizing, or kinesiophobia. These findings confirm the importance of the combined predictive ability of COMT with psychological distress and reveal other novel combinations of genetic and psychological factors that may merit additional investigation in other pain cohorts. Interactions between genetic and psychological factors were investigated as predictors of different exercise-induced shoulder pain phenotypes. The strongest statistical evidence was for interactions between the COMT diplotype and pain catastrophizing (for upper extremity disability) or depressive symptoms (for pain duration). Other novel genetic and psychological combinations were identified that may merit further investigation.


Stress-Induced, p53-Mediated Tumor Growth Inhibition of Melanoma by Modulated Electrohyperthermia in Mouse Models without Major Immunogenic Effects.
Modulated electrohyperthermia (mEHT), an innovative complementary technique of radio-, chemo-, and targeted oncotherapy modalities, can induce tumor apoptosis and contribute to a secondary immune-mediated cancer death. Here, we tested the efficiency of high-fever range (~42 °C) mEHT on B16F10 melanoma both in cell culture and allograft models. In vivo, mEHT treatment resulted in significant tumor size reduction when repeated three times, and induced major stress response as indicated by upregulated cytoplasmic and cell membrane hsp70 levels. Despite the increased PUMA and apoptosis-inducing factor 1, and moderate rise in activated-caspase-3, apoptosis was not significant. However, phospho-H2AX indicated DNA double-strand breaks, which upregulated p53 protein and its downstream cyclin-dependent kinase inhibitors p21waf1 and p27kip. Combined in vitro treatment with mEHT and the p53 activator nutlin-3a additively reduced cell viability compared to monotherapies. Though mEHT promoted the release of damage-associated molecular pattern (DAMP) damage signaling molecules hsp70, HMGB1 and ATP to potentiate the tumor immunogenicity of melanoma allografts, it reduced MHC-I and melan-A levels in tumor cells. This might explain why the number of cytotoxic T cells was moderately reduced, while the amount of natural killer (NK) cells was mainly unchanged and only macrophages increased significantly. Our results suggest that mEHT-treatment-related tumor growth control was primarily mediated by cell-stress-induced p53, which upregulated cyclin-dependent kinase inhibitors. The downregulated tumor antigen-presenting machinery may explain the reduced cytotoxic T-cell response despite increased DAMP signaling. Decreased tumor antigen and MHC-I levels suggest that natural killer (NK) cells and macrophages were the major contributors to tumor eradication.


Artesunate versus quinine in the treatment of severe falciparum malaria in African children (AQUAMAT): an open-label, randomised trial.
Severe malaria is a major cause of childhood death and often the main reason for paediatric hospital admission in sub-Saharan Africa. Quinine is still the established treatment of choice, although evidence from Asia suggests that artesunate is associated with a lower mortality. We compared parenteral treatment with either artesunate or quinine in African children with severe malaria. This open-label, randomised trial was undertaken in 11 centres in nine African countries. Children (<15 years) with severe falciparum malaria were randomly assigned to parenteral artesunate or parenteral quinine. Randomisation was in blocks of 20, with study numbers corresponding to treatment allocations kept inside opaque sealed paper envelopes. The trial was open label at each site, and none of the investigators or trialists, apart from for the trial statistician, had access to the summaries of treatment allocations. The primary outcome measure was in-hospital mortality, analysed by intention to treat. This trial is registered, number ISRCTN50258054. 5425 children were enrolled; 2712 were assigned to artesunate and 2713 to quinine. All patients were analysed for the primary outcome. 230 (8·5%) patients assigned to artesunate treatment died compared with 297 (10·9%) assigned to quinine treatment (odds ratio [OR] stratified for study site 0·75, 95% CI 0·63-0·90; relative reduction 22·5%, 95% CI 8·1-36·9; p=0·0022). Incidence of neurological sequelae did not differ significantly between groups, but the development of coma (65/1832 [3·5%] with artesunate vs 91/1768 [5·1%] with quinine; OR 0·69 95% CI 0·49-0·95; p=0·0231), convulsions (224/2712 [8·3%] vs 273/2713 [10·1%]; OR 0·80, 0·66-0·97; p=0·0199), and deterioration of the coma score (166/2712 [6·1%] vs 208/2713 [7·7%]; OR 0·78, 0·64-0·97; p=0·0245) were all significantly less frequent in artesunate recipients than in quinine recipients. Post-treatment hypoglycaemia was also less frequent in patients assigned to artesunate than in those assigned to quinine (48/2712 [1·8%] vs 75/2713 [2·8%]; OR 0·63, 0·43-0·91; p=0·0134). Artesunate was well tolerated, with no serious drug-related adverse effects. Artesunate substantially reduces mortality in African children with severe malaria. These data, together with a meta-analysis of all trials comparing artesunate and quinine, strongly suggest that parenteral artesunate should replace quinine as the treatment of choice for severe falciparum malaria worldwide. The Wellcome Trust.


Properdin binding to complement activating surfaces depends on initial C3b deposition.
Two functions have been assigned to properdin; stabilization of the alternative convertase, C3bBb, is well accepted, whereas the role of properdin as pattern recognition molecule is controversial. The presence of nonphysiological aggregates in purified properdin preparations and experimental models that do not allow discrimination between the initial binding of properdin and binding secondary to C3b deposition is a critical factor contributing to this controversy. In previous work, by inhibiting C3, we showed that properdin binding to zymosan and Escherichia coli is not a primary event, but rather is solely dependent on initial C3 deposition. In the present study, we found that properdin in human serum bound dose-dependently to solid-phase myeloperoxidase. This binding was dependent on C3 activation, as demonstrated by the lack of binding in human serum with the C3-inhibitor compstatin Cp40, in C3-depleted human serum, or when purified properdin is applied in buffer. Similarly, binding of properdin to the surface of human umbilical vein endothelial cells or Neisseria meningitidis after incubation with human serum was completely C3-dependent, as detected by flow cytometry. Properdin, which lacks the structural homology shared by other complement pattern recognition molecules and has its major function in stabilizing the C3bBb convertase, was found to bind both exogenous and endogenous molecular patterns in a completely C3-dependent manner. We therefore challenge the view of properdin as a pattern recognition molecule, and argue that the experimental conditions used to test this hypothesis should be carefully considered, with emphasis on controlling initial C3 activation under physiological conditions.


Changes in the apical surface of chloride cells following acclimation of lampreys to seawater.
Scanning electron microscopy (SEM) was used to study the changes that occur in the morphological relationships between chloride and pavement cells in the gills during acclimation of young adult lampreys to seawater. Because chloride cells are located predominantly between lamellae and are thus obscured from view, the lamellae were removed with the use of a micromanipulator installed in a SEM. In gills of animals maintained in river water, chloride cells could then be seen to be dislike and typically to form single rows between successive lamellae. After acclimation to seawater, the apical surfaces of chloride cells lose their microvilli and change in shape from small circles to rectangles that extend the full width between successive lamellae. These changes result in an increase in the length of the paracellular pathway between chloride cells. Previous work has shown that the number of strands of the zonulae occludentes sealing this pathway declines under these conditions. This presumably leads to an increase in paracellular permeability of the gill epithelium, thereby providing the low-resistance paracellular shunt required for the passive movement of sodium into the environment during osmoregulation in seawater. The above changes are reversed by transfer of lampreys downward to 10% seawater.


Low-dose oral theophylline combined with inhaled corticosteroids for people with chronic obstructive pulmonary disease and high risk of exacerbations: a RCT.
Despite widespread use of therapies such as inhaled corticosteroids (ICSs), people with chronic obstructive pulmonary disease (COPD) continue to suffer, have reduced life expectancy and utilise considerable NHS resources. Laboratory investigations have demonstrated that at low plasma concentrations (1-5 mg/l) theophylline markedly enhances the anti-inflammatory effects of corticosteroids in COPD. To determine the clinical effectiveness and cost-effectiveness of adding low-dose theophylline to a drug regimen containing ICSs in people with COPD at high risk of exacerbation. A multicentre, pragmatic, double-blind, randomised, placebo-controlled clinical trial. The trial was conducted in 121 UK primary and secondary care sites. People with COPD [i.e. who have a forced expiratory volume in 1 second (FEV1)/forced vital capacity (FVC) of < 0.7] currently on a drug regimen including ICSs with a history of two or more exacerbations treated with antibiotics and/or oral corticosteroids (OCSs) in the previous year. Participants were randomised (1 : 1) to receive either low-dose theophylline or placebo for 1 year. The dose of theophylline (200 mg once or twice a day) was determined by ideal body weight and smoking status. The number of participant-reported exacerbations in the 1-year treatment period that were treated with antibiotics and/or OCSs. A total of 1578 people were randomised (60% from primary care): 791 to theophylline and 787 to placebo. There were 11 post-randomisation exclusions. Trial medication was prescribed to 1567 participants: 788 in the theophylline arm and 779 in the placebo arm. Participants in the trial arms were well balanced in terms of characteristics. The mean age was 68.4 [standard deviation (SD) 8.4] years, 54% were male, 32% smoked and mean FEV1 was 51.7% (SD 20.0%) predicted. Primary outcome data were available for 98% of participants: 772 in the theophylline arm and 764 in the placebo arm. There were 1489 person-years of follow-up data. The mean number of exacerbations was 2.24 (SD 1.99) for participants allocated to theophylline and 2.23 (SD 1.97) for participants allocated to placebo [adjusted incidence rate ratio (IRR) 0.99, 95% confidence interval (CI) 0.91 to 1.08]. Low-dose theophylline had no significant effects on lung function (i.e. FEV1), incidence of pneumonia, mortality, breathlessness or measures of quality of life or disease impact. Hospital admissions due to COPD exacerbation were less frequent with low-dose theophylline (adjusted IRR 0.72, 95% CI 0.55 to 0.94). However, 39 of the 51 excess hospital admissions in the placebo group were accounted for by 10 participants having three or more exacerbations. There were no differences in the reporting of theophylline side effects between the theophylline and placebo arms. A higher than expected percentage of participants (26%) ceased trial medication; this was balanced between the theophylline and placebo arms and mitigated by over-recruitment (n = 154 additional participants were recruited) and the high rate of follow-up. The limitation of not using documented exacerbations is addressed by evidence that patient recall is highly reliable and the results of a small within-trial validation study. For people with COPD at high risk of exacerbation, the addition of low-dose oral theophylline to a drug regimen that includes ICSs confers no overall clinical or health economic benefit. This result was evident from the intention-to-treat and per-protocol analyses. To promote consideration of the findings of this trial in national and international COPD guidelines. Current Controlled Trials ISRCTN27066620. This project was funded by the National Institute for Health Research (NIHR) Health Technology Assessment programme and will be published in full in Health Technology Assessment; Vol. 23, No. 37. See the NIHR Journals Library website for further project information.


Transcription of Interleukin-8: How Altered Regulation Can Affect Cystic Fibrosis Lung Disease.
Interleukin-8 (IL-8) is a neutrophil chemokine that is encoded on the CXCL8 gene. Normally CXCL8 expression is repressed due to histone deacetylation, octamer-1 binding to the promoter and the inhibitory effect of nuclear factor-κB repressing factor (NRF). However, in response to a suitable stimulus, the human CXCL8 gene undergoes transcription due to its inducible promoter that is regulated by the transcription factors nuclear factor-κB (NF-κB), activating protein (AP-1), CAAT/enhancer-binding protein β (C/EBPβ, also known as NF-IL-6), C/EBP homologous protein (CHOP) and cAMP response element binding protein (CREB). CXCL8 mRNA is then stabilised by the activity of p38 mitogen-activated protein kinase (p38 MAPK). Cystic fibrosis (CF) lung disease is characterised by a neutrophil-dominated airway inflammatory response. A major factor contributing to the large number of neutrophils is the higher than normal levels of IL-8 that are present within the CF lung. Infection and inflammation, together with intrinsic alterations in CF airway cells are responsible for the abnormally high intrapulmonary levels of IL-8. Strategies to inhibit aberrantly high CXCL8 expression hold therapeutic potential for CF lung disease.


The reckoning: The return of genomic results to 1444 participants across the eMERGE3 Network.
The goal of Electronic Medical Records and Genomics (eMERGE) Phase III Network was to return actionable sequence variants to 25,084 consenting participants from 10 different health care institutions across the United States. The purpose of this study was to evaluate system-based issues relating to the return of results (RoR) disclosure process for clinical grade research genomic tests to eMERGE3 participants. RoR processes were developed and approved by each eMERGE institution's internal review board. Investigators at each eMERGE3 site were surveyed for RoR processes related to the participant's disclosure of pathogenic or likely pathogenic variants and engagement with genetic counseling. Standard statistical analysis was performed. Of the 25,084 eMERGE participants, 1444 had a pathogenic or likely pathogenic variant identified on the eMERGEseq panel of 67 genes and 14 single nucleotide variants. Of these, 1077 (74.6%) participants had results disclosed, with 562 (38.9%) participants provided with variant-specific genetic counseling. Site-specific processes that either offered or required genetic counseling in their RoR process had an effect on whether a participant ultimately engaged with genetic counseling (P = .0052). The real-life experience of the multiarm eMERGE3 RoR study for returning actionable genomic results to consented research participants showed the impact of consent, method of disclosure, and genetic counseling on RoR.


Biofilm development during the start-up of a sulfate-reducing down-flow fluidized bed reactor at different COD/SO4(2-) ratios and HRT.
In sulfate-reducing reactors, it has been reported that the sulfate removal efficiency increases when the COD/SO4(2-) ratio is increased. The start-up of a down-flow fluidized bed reactor constitutes an important step to establish a microbial community in the biofilm able to survive under the operational bioreactor conditions in order to achieve effective removal of both sulfate and organic matter. In this work the influence of COD/SO4(2-) ratio and HRT in the development of a biofilm during reactor start-up (35 days) was studied. The reactor was inoculated with 1.6 g VSS/L of granular sludge, ground low density polyethylene was used as support material; the feed consisted of mineral medium at pH 5.5 containing 1 g COD/L (acetate:lactate, 70:30) and sodium sulfate. Four experiments were conducted at HRT of 1 or 2 days and COD/SO4(2-) ratio of 0.67 or 2.5. The results obtained indicated that a COD/SO4(2-) ratio of 2.5 and HRT 2 days allowed high sulfate and COD removal (66.1 and 69.8%, respectively), whereas maximum amount of attached biomass (1.9 g SVI/L support) and highest sulfate reducing biofilm activity (10.1 g COD-H2S/g VSS-d) was achieved at HRT of 1 day and at COD/sulfate ratios of 0.67 and 2.5, respectively, which suggests that suspended biomass also played a key role in the performance of the reactors.


Knowledge and practice of occupational infection control among healthcare workers in Jamaica.
To assess the knowledge, compliance and practice among healthcare workers of occupational infection control at two hospitals in Jamaica. Employing a cross-sectional study design, medical personnel (physicians and nurses) at two hospitals in Jamaica, were studied, utilizing a structured questionnaire consisting of 14 items to collect the data. Participants considered the following fluids, not blood stained, high risk for HIV transmission: breast milk (79%), saliva (14%), urine (27%), pleural fluid (53%), CSF (55%), synovial fluid (37%), faeces (27%), peritoneal fluid (53%) and vomitus (21%). The respondents estimated the risk of transmission of infection after a needlestick injury from a patient with: HIV, mean 22.5%, HB, 34% and HCV, 26%. Needles for drawing blood were identified as having the highest risk for transmission of infections in 63%. The following precautions were adhered to all the time: wearing gloves (38%), not re-sheathing needles (22%), not passing needles directly to others (70%), properly disposing of sharps (86%) and regarding patients' blood and other high risk fluid as potentially infected (62%). Post exposure, 43% indicated bleeding/squeezing the NSI site as the initial first-aid procedure, washing with soap and water (29%) and irrigating the area with water (20%). Healthcare workers are aware of the risk of transmission of infection, however compliance with universal precautions was inadequate. An improvement in knowledge and practice with clear guidelines are needed and a comprehensive programme to educate HCWs regarding compliance with universal precautions is urgently required.


Do graphic health warning labels have an impact on adolescents' smoking-related beliefs and behaviours?
To assess the impact of the introduction of graphic health warning labels on cigarette packets on adolescents at different smoking uptake stages. School-based surveys conducted in the year prior to (2005) and approximately 6 months after (2006) the introduction of the graphic health warnings. The 2006 survey was conducted after a TV advertising campaign promoting two new health warnings. Secondary schools in greater metropolitan Melbourne, Australia. Students in year levels 8-12: 2432 students in 2005, and 2050 in 2006, participated. Smoking uptake stage, intention to smoke, reported exposure to cigarette packs, knowledge of health effects of smoking, cognitive processing of warning labels and perceptions of cigarette pack image. At baseline, 72% of students had seen cigarette packs in the previous 6 months, while at follow-up 77% had seen packs and 88% of these had seen the new warning labels. Cognitive processing of warning labels increased, with students more frequently reading, attending to, thinking and talking about warning labels at follow-up. Experimental and established smokers thought about quitting and forgoing cigarettes more at follow-up. At follow-up intention to smoke was lower among those students who had talked about the warning labels and had forgone cigarettes. Graphic warning labels on cigarette packs are noticed by the majority of adolescents, increase adolescents' cognitive processing of these messages and have the potential to lower smoking intentions. Our findings suggest that the introduction of graphic warning labels may help to reduce smoking among adolescents.


Dwarfism and impaired gut development in insulin-like growth factor II mRNA-binding protein 1-deficient mice.
Insulin-like growth factor II mRNA-binding protein 1 (IMP1) belongs to a family of RNA-binding proteins implicated in mRNA localization, turnover, and translational control. Mouse IMP1 is expressed during early development, and an increase in expression occurs around embryonic day 12.5 (E12.5). To characterize the physiological role of IMP1, we generated IMP1-deficient mice carrying a gene trap insertion in the Imp1 gene. Imp1(-/-) mice were on average 40% smaller than wild-type and heterozygous sex-matched littermates. Growth retardation was apparent from E17.5 and remained permanent into adult life. Moreover, Imp1(-/-) mice exhibited high perinatal mortality, and only 50% were alive 3 days after birth. In contrast to most other organs, intestinal epithelial cells continue to express IMP1 postnatally, and Imp1(-/-) mice exhibited impaired development of the intestine, with small and misshapen villi and twisted colon crypts. Analysis of target mRNAs and global expression profiling at E12.5 indicated that Igf2 translation was downregulated, whereas the postnatal intestine showed reduced expression of transcripts encoding extracellular matrix components, such as galectin- 1, lumican, tenascin-C, procollagen transcripts, and the Hsp47 procollagen chaperone. Taken together, the results demonstrate that IMP1 is essential for normal growth and development. Moreover, IMP1 may facilitate intestinal morphogenesis via regulation of extracellular matrix formation.


The star coupler: a unique interconnection component for multimode optical waveguide communications systems.
The use of optical waveguides in communications systems may require the interconnection of many interactive terminals. The preferred configuration for passive interconnection in optical waveguide systems is shown to be a configuration termed the Star system. The central interconnecting component, the Star coupler, is described, and the performance of a system incorporating it is compared to that of an equivalent system using a common optical bus and Tee couplers. The Star system is shown to have lower terminal-to-terminal loss, easier upgradability, enhanced reliability and survivability, inherent bidirectionality, and inherent equivalence of all terminals. Construction of two Star couplers is described, and measurements of their performance characteristics are reported.


Study of the most relevant spin-orbit coupling descriptions of magnetic excitations in a series of lanthanide complexes.
We present a number of computationally cost-effective approaches to calculate magnetic excitations (i.e. crystal field energies and magnetic anisotropies in the lowest spin-orbit multiplet) in lanthanide complexes. In particular, we focus on the representation of the spin-orbit coupling term of the molecular Hamiltonian, which has been implemented within the quantum chemistry package CERES using various approximations to the Breit-Pauli Hamiltonian. The approximations include the (i) bare one-electron approximation, (ii) atomic mean field and molecular mean field approximations of the two-electron term, (iii) full representation of the Breit-Pauli Hamiltonian. Within the framework of the CERES implementation, the spin-orbit Hamiltonian is always fully diagonalized together with the electron repulsion Hamiltonian (CASCI-SO) on the full basis of Slater determinants arising within the 4f ligand field space. For the first time, we make full use of the Cholesky decomposition of two-electron spin-orbit integrals to speed up the calculation of the two-electron spin-orbit operator. We perform an extensive comparison of the different approximations on a set of lanthanide complexes varying both the lanthanide ion and the ligands. Surprisingly, while our results confirm the need of at least a mean field approach to accurately describe the spin-orbit coupling interaction within the ground Russell-Saunders term, we find that the simple bare one-electron spin-orbit Hamiltonian performs reasonably well to describe the crystal field split energies and g tensors within the ground spin-orbit multiplet, which characterize all the magnetic excitations responsible for lanthanide-based single-molecule magnetism.


Treatment of trigeminal ganglion neurons in vitro with NGF, GDNF or BDNF: effects on neuronal survival, neurochemical properties and TRPV1-mediated neuropeptide secretion.
Nerve growth factor (NGF), glial cell line-derived neurotrophic factor (GDNF) and brain-derived neurotrophic factor (BDNF) all play important roles in the development of the peripheral sensory nervous system. Additionally, these growth factors are proposed to modulate the properties of the sensory system in the adult under pathological conditions brought about by nerve injury or inflammation. We have examined the effects of NGF, GDNF and BDNF on adult rat trigeminal ganglion (TG) neurons in culture to gain a better understanding of how these growth factors alter the cytochemical and functional phenotype of these neurons, with special attention to properties associated with nociception. Compared with no growth factor controls, GDNF, at 1 and 100 ng/ml, significantly increased by nearly 100% the number of neurons in culture at 5 days post-plating. A significant, positive, linear trend of increasing neuron number as a function of BDNF concentration was observed, also peaking at nearly 100%. NGF treatment was without effect. Chronic treatment with NGF and GDNF significantly and concentration-dependently increased 100 nM capsaicin (CAP)-evoked calcitonin gene-related peptide (CGRP) release, reaching approximately 300% at the highest concentration tested (100 ng/ml). Also, NGF and GDNF each augmented anandamide (AEA)- and arachidonyl-2-chloroethylamide (ACEA)-evoked CGRP release, while BDNF was without effect. Utilizing immunohistochemistry to account for the proportions of TRPV1- or CGRP-positive neurons under each growth factor treatment condition and then standardizing evoked CGRP release to these proportions, we observed that NGF was much more effective in enhancing CAP- and 50 mM K+-evoked CGRP release than was GDNF. Furthermore, NGF and GDNF each altered the concentration-response function for CAP- and AEA-evoked CGRP release, increasing the Emax without altering the EC50 for either compound. Taken together, our results illustrate that NGF, GDNF and BDNF differentially alter TG sensory neuron survival, neurochemical properties and TRPV1-mediated neuropeptide release in culture. In particular, our findings suggest that GDNF and NGF differentially modulate TRPV1-mediated neuropeptide secretion sensitivity, with NGF having a much greater effect on a per neuron basis than GDNF. These findings are discussed in relation to possible therapeutic roles for growth factors or their modulators in pathological pain states, especially as these relate to the trigeminal system.


Novel insights into breast cancer genetic variance through RNA sequencing.
Using RNA sequencing of triple-negative breast cancer (TNBC), non-TBNC and HER2-positive breast cancer sub-types, here we report novel expressed variants, allelic prevalence and abundance, and coexpression with other variation, and splicing signatures. To reveal the most prevalent variant alleles, we overlaid our findings with cancer- and population-based datasets and validated a subset of novel variants of cancer-related genes: ESRP2, GBP1, TPP1, MAD2L1BP, GLUD2 and SLC30A8. As a proof-of-principle, we demonstrated that a rare substitution in the splicing coordinator ESRP2 (R353Q) impairs its ability to bind to its substrate FGFR2 pre-mRNA. In addition, we describe novel SNPs and INDELs in cancer relevant genes with no prior reported association of point mutations with cancer, such as MTAP and MAGED1. For the first time, this study illustrates the power of RNA-sequencing in revealing the variation landscape of breast transcriptome and exemplifies analytical strategies to search regulatory interactions among cancer relevant molecules.


Rapid immunoprecipitation mass spectrometry of endogenous proteins (RIME) for analysis of chromatin complexes.
Rapid immunoprecipitation mass spectrometry of endogenous protein (RIME) is a method that allows the study of protein complexes, in particular chromatin and transcription factor complexes, in a rapid and robust manner by mass spectrometry (MS). The method can be used in parallel with chromatin immunoprecipitation-sequencing (ChIP-seq) experiments to provide information on both the cistrome and interactome for a given protein. The method uses formaldehyde fixation to stabilize protein complexes. By using antibodies against the endogenous target, the cross-linked complex is immunoprecipitated, rigorously washed, and then digested into peptides while avoiding antibody contamination (on-bead digestion). By using this method, MS identification of the target protein and several dozen interacting proteins is possible using a 100-min LC-MS/MS run. The protocol does not require substantial proteomics expertise, and it typically takes 2-3 d from the collection of material to results.


The T-box transcription factor Eomesodermin governs haemogenic competence of yolk sac mesodermal progenitors.
Extra-embryonic mesoderm (ExM)-composed of the earliest cells that traverse the primitive streak-gives rise to the endothelium as well as haematopoietic progenitors in the developing yolk sac. How a specific subset of ExM becomes committed to a haematopoietic fate remains unclear. Here we demonstrate using an embryonic stem cell model that transient expression of the T-box transcription factor Eomesodermin (Eomes) governs haemogenic competency of ExM. Eomes regulates the accessibility of enhancers that the transcription factor stem cell leukaemia (SCL) normally utilizes to specify primitive erythrocytes and is essential for the normal development of Runx1+ haemogenic endothelium. Single-cell RNA sequencing suggests that Eomes loss of function profoundly blocks the formation of blood progenitors but not specification of Flk-1+ haematoendothelial progenitors. Our findings place Eomes at the top of the transcriptional hierarchy regulating early blood formation and suggest that haemogenic competence is endowed earlier during embryonic development than was previously appreciated.


High-relaxivity superparamagnetic iron oxide nanoworms with decreased immune recognition and long-circulating properties.
One of the core issues of nanotechnology involves masking the foreignness of nanomaterials to enable in vivo longevity and long-term immune evasion. Dextran-coated superparamagnetic iron oxide nanoparticles are very effective magnetic resonance imaging (MRI) contrast agents, and strategies to prevent immune recognition are critical for their clinical translation. Here we prepared 20 kDa dextran-coated SPIO nanoworms (NWs) of 250 nm diameter and a high molar transverse relaxivity rate R2 (∼400 mM(-1) s(-1)) to study the effect of cross-linking-hydrogelation with 1-chloro-2,3-epoxypropane (epichlorohydrin) on the immune evasion both in vitro and in vivo. Cross-linking was performed in the presence of different concentrations of NaOH (0.5 to 10 N) and different temperatures (23 and 37 °C). Increasing NaOH concentration and temperature significantly decrease the binding of anti-dextran antibody and dextran-binding lectin conconavalin A to the NWs. The decrease in dextran immunoreactivity correlated with the decrease in opsonization by complement component 3 (C3) and with the decrease in the binding of the lectin pathway factor MASP-2 in mouse serum, suggesting that cross-linking blocks the lectin pathway of complement. The decrease in C3 opsonization correlated with the decrease in NW uptake by murine peritoneal macrophages. Optimized NWs demonstrated up to 10 h circulation half-life in mice and minimal uptake by the liver, while maintaining the large 250 nm size in the blood. We demonstrate that immune recognition of large iron oxide nanoparticles can be efficiently blocked by chemical cross-linking-hydrogelation, which is a promising strategy to improve safety and bioinertness of MRI contrast agents.


A Network Model of Local Field Potential Activity in Essential Tremor and the Impact of Deep Brain Stimulation.
Essential tremor (ET), a movement disorder characterised by an uncontrollable shaking of the affected body part, is often professed to be the most common movement disorder, affecting up to one percent of adults over 40 years of age. The precise cause of ET is unknown, however pathological oscillations of a network of a number of brain regions are implicated in leading to the disorder. Deep brain stimulation (DBS) is a clinical therapy used to alleviate the symptoms of a number of movement disorders. DBS involves the surgical implantation of electrodes into specific nuclei in the brain. For ET the targeted region is the ventralis intermedius (Vim) nucleus of the thalamus. Though DBS is effective for treating ET, the mechanism through which the therapeutic effect is obtained is not understood. To elucidate the mechanism underlying the pathological network activity and the effect of DBS on such activity, we take a computational modelling approach combined with electrophysiological data. The pathological brain activity was recorded intra-operatively via implanted DBS electrodes, whilst simultaneously recording muscle activity of the affected limbs. We modelled the network hypothesised to underlie ET using the Wilson-Cowan approach. The modelled network exhibited oscillatory behaviour within the tremor frequency range, as did our electrophysiological data. By applying a DBS-like input we suppressed these oscillations. This study shows that the dynamics of the ET network support oscillations at the tremor frequency and the application of a DBS-like input disrupts this activity, which could be one mechanism underlying the therapeutic benefit.


Kenyan Isolates of Puccinia graminis f. sp. tritici from 2008 to 2014: Virulence to SrTmp in the Ug99 Race Group and Implications for Breeding Programs.
Frequent emergence of new variants in the Puccinia graminis f. sp. tritici Ug99 race group in Kenya has made pathogen survey a priority. We analyzed 140 isolates from 78 P. graminis f. sp. tritici samples collected in Kenya between 2008 and 2014 and identified six races, including three not detected prior to 2013. Genotypic analysis of 20 isolates from 2013 and 2014 collections showed that the new races TTHST, TTKTK, and TTKTT belong to the Ug99 race group. International advanced breeding lines were evaluated against an isolate of TTKTT (Sr31, Sr24, and SrTmp virulence) at the seedling stage. From 169 advanced lines from Kenya, 23% of lines with resistance to races TTKSK and TTKST were susceptible to TTKTT and, from two North American regional nurseries, 44 and 91% of resistant lines were susceptible. Three lines with combined resistance genes were developed to facilitate pathogen monitoring and race identification. These results indicate the increasing virulence and variability in the Kenyan P. graminis f. sp. tritici population and reveal vulnerabilities of elite germplasm to new races.


Human tumour immune evasion via TGF-β blocks NK cell activation but not survival allowing therapeutic restoration of anti-tumour activity.
Immune evasion is now recognized as a key feature of cancer progression. In animal models, the activity of cytotoxic lymphocytes is suppressed in the tumour microenvironment by the immunosuppressive cytokine, Transforming Growth Factor (TGF)-β. Release from TGF-β-mediated inhibition restores anti-tumour immunity, suggesting a therapeutic strategy for human cancer. We demonstrate that human natural killer (NK) cells are inhibited in a TGF-β dependent manner following chronic contact-dependent interactions with tumour cells in vitro. In vivo, NK cell inhibition was localised to the human tumour microenvironment and primary ovarian tumours conferred TGF-β dependent inhibition upon autologous NK cells ex vivo. TGF-β antagonized the interleukin (IL)-15 induced proliferation and gene expression associated with NK cell activation, inhibiting the expression of both NK cell activation receptor molecules and components of the cytotoxic apparatus. Interleukin-15 also promotes NK cell survival and IL-15 excluded the pro-apoptotic transcription factor FOXO3 from the nucleus. However, this IL-15 mediated pathway was unaffected by TGF-β treatment, allowing NK cell survival. This suggested that NK cells in the tumour microenvironment might have their activity restored by TGF-β blockade and both anti-TGF-β antibodies and a small molecule inhibitor of TGF-β signalling restored the effector function of NK cells inhibited by autologous tumour cells. Thus, TGF-β blunts NK cell activation within the human tumour microenvironment but this evasion mechanism can be therapeutically targeted, boosting anti-tumour immunity.


The potential of positron emission tomography for intratreatment dynamic lung tumor tracking: a phantom study.
This study aims to evaluate the potential and feasibility of positron emission tomography for dynamic lung tumor tracking during radiation treatment. The authors propose a center of mass (CoM) tumor tracking algorithm using gated-PET images combined with a respiratory monitor and investigate the geometric accuracy of the proposed algorithm. The proposed PET dynamic lung tumor tracking algorithm estimated the target position information through the CoM of the segmented target volume on gated PET images reconstructed from accumulated coincidence events. The information was continuously updated throughout a scan based on the assumption that real-time processing was supported (actual processing time at each frame ≈ 10 s). External respiratory motion and list-mode PET data were acquired from a phantom programmed to move with measured respiratory traces (external respiratory motion and internal target motion) from human subjects, for which the ground truth target position was known as a function of time. The phantom was cylindrical with six hollow sphere targets (10, 13, 17, 22, 28, and 37 mm in diameter). The measured respiratory traces consisted of two sets: (1) 1D-measured motion from ten healthy volunteers and (2) 3D-measured motion from four lung cancer patients. The authors evaluated the geometric accuracy of the proposed algorithm by quantifying estimation errors (Euclidean distance) between the actual motion of targets (1D-motion and 3D-motion traces) and CoM trajectories estimated by the proposed algorithm as a function of time. The time-averaged error of 1D-motion traces over all trajectories of all targets was 1.6 mm. The error trajectories decreased with time as coincidence events were accumulated. The overall error trajectory of 1D-motion traces converged to within 2 mm in approximately 90 s. As expected, more accurate results were obtained for larger targets. For example, for the 37 mm target, the average error over all 1D-motion traces was 1.1 mm; and for the 10 mm target, the average error over all 1D-motion traces was 2.8 mm. The overall time-averaged error of 3D-motion traces was 1.6 mm, which was comparable to that of the 1D-motion traces. There were small variations in the errors between the 3D-motion traces, although the motion trajectories were very different. The accuracy of the estimates was consistent for all targets except for the smallest. The authors developed an algorithm for dynamic lung tumor tracking using list-mode PET data and a respiratory motion signal, and demonstrated proof-of-principle for PET-guided lung tumor tracking. The overall tracking error in phantom studies is less than 2 mm.


Bath thermal waters in the treatment of knee osteoarthritis: a randomized controlled clinical trial.
Osteoarthritis is a degenerative disease associated with pain, reduced range of motion, and impaired function. Balneotherapy or bathing in thermal or mineral waters is used as a non-invasive treatment for various rheumatic diseases. To evaluate the effectiveness of hot sulfurous and non-sulfurous waters in the treatment of knee osteoarthritis. A randomized, assessor-blind, controlled trial. A spa resort. One hundred and forty patients of both genders, mean age of 64.8±8.9 years, with knee osteoarthritis and chronic knee pain. Patients were randomized into three groups: the sulfurous water (SW) group (N.=47), non-sulfurous water (NSW) group (N.=50), or control group (N.=43) who received no treatment. Patients were not blinded to treatment allocation. Treatment groups received 30 individual thermal baths (three 20-minute baths a week for 10 weeks) at 37-39 °C. The outcome measures were pain (visual analog scale, VAS), physical function (Western Ontario and McMaster Universities Osteoarthritis Index, WOMAC; Lequesne Algofunctional Index, LAFI; Stanford Health Assessment Questionnaire, HAQ), and use of pain medication. Patients were assessed before treatment (T1), at treatment endpoint (T2), and two months post-intervention (T3). Intra- and intergroup comparisons were performed at a significance level of 0.05 (P<0.05). A significant decrease in VAS pain scores (pain during movement, at rest, and at night) and use of pain medication, and improvement in WOMAC, LAFI and HAQ scores were observed from baseline to T2 and T3 within treatment groups (P<0.001), and between controls and both treatment groups at T2 and T3 (P<0.001). No significant differences in these variables were observed between treatment groups at T2, but patients in the SW group reported less pain and better functional status than those in the NSW group at T3, showing a lasting effect of sulfurous water baths. Both therapeutic methods were effective in the treatment of knee osteoarthritis; however, sulfurous baths yielded longer-lasting effects than non-sulfurous water baths. Baths in thermal waters, especially those in sulfurous waters, are effective in reducing pain and improving physical function in patients with knee osteoarthritis.


Fractionated Stereotactic Radiotherapy for Pituitary Adenomas: Single-Center Experience in 75 Consecutive Patients.
Early results of postoperative fractionated stereotactic radiotherapy (FSRT) for functional and nonfunctional pituitary adenomas appear promising, but the majority of available evidence draws from small series with insufficient follow-up data to draw meaningful conclusions. To evaluate the long-term outcomes of a large series of patients undergoing FSRT for both functional and nonfunctional pituitary adenomas with the Novalis system (BrainLAB, Heimstetten, Germany). Chart data for 75 consecutive patients undergoing FSRT for a pituitary tumor (21 functional and 54 nonfunctional adenomas) at our institution between January 2004 and June 2013 were reviewed. Radiographic progression-free survival was 100% over a mean of 47.8 months of radiographic follow-up (range, 12.0-131.2 months). Hormonal normalization was seen in 69.2% of patients with functional adenomas after FSRT, whereas 30.8% experienced partial hormonal control. Mild, grade I acute adverse effects were observed during radiotherapy treatment in 36 patients (48%), and objective, persistent worsening of vision occurred in a single patient (1.5%) after FSRT. New hormonal deficits were seen in 28.0% of patients after FSRT. Radiographic responses were inversely related to tumor volume. FSRT delivers radiographic and functional outcomes similar to those seen with stereotactic radiosurgery and conventional radiotherapy with less resultant toxicity. FSRT is most beneficial for smaller tumors (those <3 cm in diameter). EBRT, external beam radiotherapyFSRT, fractionated stereotactic radiotherapyOR, odds ratioPTV, planning target volumeSRS, stereotactic radiosurgery.


Physical plasma therapy accelerates wound re-epithelialisation and enhances extracellular matrix formation in cutaneous skin grafts.
Skin grafting is a surgical method of cutaneous reconstruction, which provides volumetric replacement in wounds unable to heal by primary intention. Clinically, full-thickness skin grafts (FTSGs) are placed in aesthetically sensitive and mechanically demanding areas such as the hands, face, and neck. Complete or partial graft failure is the primary complication associated with this surgical procedure. Strategies aimed at improving the rate of skin graft integration will reduce the incidence of graft failure. Cold atmospheric plasma (CAP) is an emerging technology offering innovative clinical applications. The aim of this study was to test the therapeutic potential of CAP to improve wound healing and skin graft integration into the recipient site. In vitro models that mimic wound healing were used to investigate the ability of CAP to enhance cellular migration, a key factor in cutaneous tissue repair. We demonstrated that CAP enhanced the migration of epidermal keratinocytes and dermal fibroblasts. This increased cellular migration was possibly induced by the low dose of reactive oxygen and nitrogen species produced by CAP. Using a mouse model of burn wound reconstructed with a full-thickness skin graft, we showed that wounds treated with CAP healed faster than did control wounds. Immunohistochemical wound analysis showed that CAP treatment enhanced the expression of the dermal-epidermal junction components, which are vital for successful skin graft integration. CAP treatment was characterised by increased levels of Tgfbr1 mRNA and collagen I protein in vivo, suggesting enhanced wound maturity and extracellular matrix deposition. Mechanistically, we show that CAP induced the activation of the canonical SMAD-dependent TGF-β1 pathway in primary human dermal fibroblasts, which may explain the increased collagen I synthesis in vitro. These studies revealed that CAP improved wound repair and skin graft integration via mechanisms involving extracellular matrix formation. CAP offers a novel approach for treating cutaneous wounds and skin grafts. © 2020 The Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.


Lysopine and Octopine Promote Crown-Gall Tumor Growth in vivo.
Growth of tumors induced on primary leaves of bean plants by Agrobacterium tumefaciens was increased by the addition of lysopine and octopine. A detectable response was observed when as little as 1 microgram of these compounds was added per leaf, and the mean volume of the tumors was increased two- to threefold when greater amounts were applied. The specificity of the response and the unique association of these compounds with the tumors suggest that endogenous lysopine and octopine contribute to the growth characteristics of these tumors.


Extracellular Signal-Regulated Kinases 1 and 2 Phosphorylate Gab2 To Promote a Negative-Feedback Loop That Attenuates Phosphoinositide 3-Kinase/Akt Signaling.
The scaffolding adapter protein Gab2 (Grb2-associated binder) promotes cell proliferation, survival, and motility by engaging several signaling pathways downstream of growth factor and cytokine receptors. In particular, Gab2 plays essential roles in mast cells, as it is required for phosphoinositide 3-kinase (PI3K) activation in response to Kit and the high-affinity IgE receptor. While the positive role of Gab2 in PI3K signaling is well documented, very little is known about the mechanisms that attenuate its function. Here we show that Gab2 becomes phosphorylated on multiple proline-directed sites upon stimulation of the Ras/extracellular signal-regulated kinase (ERK) signaling pathway. We demonstrate that ERK1 and ERK2 interact with Gab2 via a novel docking motif, which is required for subsequent Gab2 phosphorylation in response to ERK1/2 activation. We identified four ERK1/2-dependent phosphorylation sites in Gab2 that prevent the recruitment of the p85 regulatory subunit of PI3K. Using bone marrow-derived mast cells to study Gab2-dependent signaling, we found that the inhibition of ERK1/2 activity promotes Akt signaling in response to Kit and the high-affinity IgE receptor. Together, our results indicate that ERK1/2 participates in a negative-feedback loop that attenuates PI3K/Akt signaling in response to various agonists.


Pancreatic Cancer: A Review.
Pancreatic ductal adenocarcinoma (PDAC) is a relatively uncommon cancer, with approximately 60 430 new diagnoses expected in 2021 in the US. The incidence of PDAC is increasing by 0.5% to 1.0% per year, and it is projected to become the second-leading cause of cancer-related mortality by 2030. Effective screening is not available for PDAC, and most patients present with locally advanced (30%-35%) or metastatic (50%-55%) disease at diagnosis. A multidisciplinary management approach is recommended. Localized pancreas cancer includes resectable, borderline resectable (localized and involving major vascular structures), and locally advanced (unresectable) disease based on the degree of arterial and venous involvement by tumor, typically of the superior mesenteric vessels. For patients with resectable disease at presentation (10%-15%), surgery followed by adjuvant chemotherapy with FOLFIRINOX (fluorouracil, irinotecan, leucovorin, oxaliplatin) represents a standard therapeutic approach with an anticipated median overall survival of 54.4 months, compared with 35 months for single-agent gemcitabine (stratified hazard ratio for death, 0.64 [95% CI, 0.48-0.86]; P = .003). Neoadjuvant systemic therapy with or without radiation followed by evaluation for surgery is an accepted treatment approach for resectable and borderline resectable disease. For patients with locally advanced and unresectable disease due to extensive vascular involvement, systemic therapy followed by radiation is an option for definitive locoregional disease control. For patients with advanced (locally advanced and metastatic) PDAC, multiagent chemotherapy regimens, including FOLFIRINOX, gemcitabine/nab-paclitaxel, and nanoliposomal irinotecan/fluorouracil, all have a survival benefit of 2 to 6 months compared with a single-agent gemcitabine. For the 5% to 7% of patients with a BRCA pathogenic germline variant and metastatic PDAC, olaparib, a poly (adenosine diphosphate [ADB]-ribose) polymerase inhibitor, is a maintenance option that improves progression-free survival following initial platinum-based therapy. Approximately 60 000 new cases of PDAC are diagnosed per year, and approximately 50% of patients have advanced disease at diagnosis. The incidence of PDAC is increasing. Currently available cytotoxic therapies for advanced disease are modestly effective. For all patients, multidisciplinary management, comprehensive germline testing, and integrated supportive care are recommended.


Distraction osteogenesis after acute limb-shortening for segmental tibial defects. Comparison of a monofocal and a bifocal technique in rabbits.
Segmental bone defects can be treated with immediate limb-shortening followed by monofocal or bifocal distraction osteogenesis. In the present study, the efficacy of monofocal distraction osteogenesis was compared with that of bifocal distraction osteogenesis in a rabbit model. Twenty-four skeletally mature New Zealand White rabbits were divided into two equal groups: one group had monofocal distraction osteosynthesis, and the other had bifocal distraction osteosynthesis. In both groups, a one-centimeter-long segment of bone was resected from the midpart of the tibial shaft. In the monofocal reconstruction group, the limb was immediately shortened to close the segmental defect and the defect was allowed to heal for ten days. Lengthening was then begun at this site, with use of a specially designed external fixator, at a rate of 0.5 millimeter per twelve hours. In the bifocal reconstruction group, the segmental defect was closed immediately and the fragments were fixed with microplates. A subperiosteal osteotomy was performed proximal to the tibiofibular junction, and lengthening was performed at the site of the osteotomy. The animals in both groups were killed twenty days after the lengthening was completed. New-bone formation then was evaluated with use of radiographs, densitometry, biomechanical testing, and histological and histomorphometric analysis. Osseous consolidation occurred in all but one of the animals. Biomechanical testing demonstrated that the tibiae that had been treated with use of the simple monofocal reconstruction technique tended to have greater torsional stiffness (p = 0.14) and strength (p = 0.09). Follow-up radiographs revealed that both groups had a significant decrease in radiolucent area (p < 0.05), which occurred at essentially the same rate after lengthening. No significant differences were found between the groups with respect to new-bone mineral density, new-bone area, or the amount of callus. Thus, after resection of a diaphyseal bone segment comprising 10 percent of the original length of the tibia and acute shortening, limb reconstruction was completed successfully through distraction osteogenesis with use of either a monofocal or a bifocal technique in rabbits. In the present study, both monofocal and bifocal techniques of shortening and distraction osteogenesis were effective for the reconstruction of segmental bone defects. Under some conditions, the monofocal method may provide a simpler means of treating such defects. Damage to the soft-tissue envelope as well as venous and lymphatic stasis impose limits on the amount of limb-shortening that can be achieved with use of the monofocal method and also influence the indications for this procedure in the clinical setting.