Population Pharmacokinetic Modeling of Tribendimidine Metabolites in Opisthorchis viverrini-Infected Adults.
There is a pressing need for alternative treatments against the liver fluke Opisthorchis viverrini Oral tribendimidine is a promising candidate, but its population pharmacokinetic properties are unknown. Two phase IIa trials were conducted in Laos in O. viverrini-infected adults receiving single oral doses of 25 to 600 mg tribendimidine administered as different formulations in each study (study 1 used 200-mg tablets, and study 2 used 50-mg tablets). Venous whole blood, plasma, and capillary dried blood spots were sampled frequently from 68 adults, and concentrations of the tribendimidine metabolites dADT (deacetylated amidantel) and adADT (acetylated dADT) were measured. Population pharmacokinetics were assessed by using nonlinear mixed-effects modeling. The relationship between drug exposure and cure (assessed at 21 days posttreatment) was evaluated by using univariable logistic regression. A six-transit compartment absorption model with a one-disposition compartment for each metabolite described the data well. Compared to the 50-mg formulation (study 2), the 200-mg formulation (study 1) had a 40.1% higher mean transit absorption time, a 113% higher dADT volume of distribution, and a 364% higher adADT volume of distribution. Each 10-year increase in age was associated with a 12.7% lower dADT clearance and a 21.2% lower adADT clearance. The highest cure rates (≥55%) were observed with doses of ≥100 mg. Higher dADT, but not adADT, peak concentrations and exposures were associated with cure (P = 0.004 and 0.003, respectively). For the first time, population pharmacokinetics of tribendimidine have been described. Known differences in the 200-mg versus 50-mg formulations were captured by covariate modeling. Further studies are needed to validate the structural model and confirm covariate relationships. (This study has been registered with the ISRCTN Registry under no. ISRCTN96948551.).


Implication of dopaminergic mechanisms in the wake-promoting effects of amphetamine: a study of D- and L-derivatives in canine narcolepsy.
Using a canine model of narcolepsy and selective DA and NE uptake inhibitors, we have recently shown that DA uptake inhibition promotes wakefulness, while NE uptake inhibition inhibits rapid eye movement sleep and cataplexy. In order to further delineate the respective roles of the dopaminergic and noradrenergic systems in the pharmacological control of symptoms of narcolepsy, we compared the potency of amphetamine isomers (D- and L-amphetamines) and a derivative (L-methamphetamine) on wakefulness and cataplexy. Their respective effects on these narcolepsy symptoms were then compared with their in vivo effects on extracellular DA levels in the caudate and NE levels in the frontal cortex during local drug perfusion in narcoleptic dogs. Polygraphic recordings demonstrated that D-amphetamine was about twice as potent as L-amphetamine, and was six times more potent than L-methamphetamine in increasing wakefulness and reducing slow-wave sleep. D-Amphetamine and L-amphetamine were equipotent in reducing rapid eye movement sleep and cataplexy, and L-methamphetamine was about half as potent as L- and D-amphetamines. D-Amphetamine was found to be more potent in increasing DA efflux than L-amphetamine, and L-methamphetamine was found to have little effect on DA efflux; there was no significant difference in the potencies of the three derivatives on NE efflux. The potencies of these amphetamines on wakefulness correlated well with DA, but not NE, efflux in the brain of narcoleptic dogs during local drug perfusion. Our current results further exemplify the importance of the DA system for the pharmacological control of electroencephalogram arousal and suggest that increased DA transmission mediates the wake-promoting effects of amphetamine-like stimulants.


Oxidative Stress in Obese Postmenopausal Women: An Additive Burden for Atherosclerosis.
Coronary Artery Disease are on the rise in the general population and is the leading cause of death in both men and women. The impact of CAD is underappreciated in younger women when compared to men. Women have unique risk factors for CAD and postmenopausal women are at higher risk of developing CAD when compared to normal menstruating women. The aim of our study was to find out the difference in oxidative stress levels between obese postmenopausal women and normal menstruating women, also to compare the same in normal weight postmenopausal women. Thirty one normal and 29 obese postmenopausal women with age more than 45 years who visited obstetrics and gynaecology outpatient department for general clinical evaluation at a tertiary care centre were recruited in this cross-sectional study. Thirty normal menstruating women were compared. Anthropometric measurements were recorded and the body mass index was calculated. Serum Malondialdehyde and superoxide dismutase was measured using a spectrophotometer. There was a significant difference in mean MDA levels in postmenopausal women (1.477 ± 0.359) when compared to normal menstruating women (0.666 ± 0.302) (p < 0.01). There was no significant difference in mean SOD levels in postmenopausal women (2.836 ± 0.899) when compared to normal menstruating women (2.986 ± 0.686) (p > 0.05). Also, there was a significant increase between mean MDA levels in obese postmenopausal women (2.48 ± 0.52) when compared to normal weight postmenopausal women (1.65 ± 0.36) (p < 0.01). There was a significant difference between mean SOD levels in obese postmenopausal women (1.36 ± 0.96) and normal weight postmenopausal women (2.56 ± 1.03) (p < 0.01). The oxidative stress was higher in obese postmenopausal women when compared to normal weight postmenopausal women and normal menstruating women.


Orbital and spin dynamics of intraband electrons in quantum rings driven by twisted light.
We theoretically investigate the effect that twisted light has on the orbital and spin dynamics of electrons in quantum rings possessing sizable Rashba spin-orbit interaction. The system Hamiltonian for such a strongly inhomogeneous light field exhibits terms which induce both spin-conserving and spin-flip processes. We analyze the dynamics in terms of the perturbation introduced by a weak light field on the Rasha electronic states, and describe the effects that the orbital angular momentum as well as the inhomogeneous character of the beam have on the orbital and the spin dynamics.


Trauma research in Iran: a report of the Sina Trauma Data Bank.
Injuries are a major health problem in developing countries. Recently, the occurrence of injuries and those related to mortality have increased dramatically in Iran. We have conducted this study to report domestic research publications in the field of trauma from 2000 to 2006 in Iran. The contents of 128 biomedical journals were analyzed. Sina Trauma Data Bank (STDB) was selected as an indicator of trauma related articles that have been published in Iranian biomedical journals. The number of trauma related articles in Iran increased during the seven year period (2000 - 2006), but the percentage of total publications was not significantly different. The mean number of authors per article was 2.7+1.3 (1 to 12) and all first authors were Iranian. Females constituted 15.4% (n=156) of first authors. Original articles were the most frequently published articles (n=871; 86.2%). Treatment of injuries and ensuing complications were the most common topics (42%) followed by war injuries (14.1%). Traffic accidents, as the leading cause of trauma death in Iran, were investigated in 27 (2.7%) articles. According to our results, it seems that more attention from researchers and health-policy makers towards trauma is necessary in our country. In addition, publishing a journal with the main theme of injury and trauma could be an effective means for increasing the proportion of trauma related articles.


Materials in embolotherapy of high-flow priapism: results and long-term follow-up.
To review our experience with embolic materials used in the selective arterial embolization of high-flow priapism and present the results of long-term follow-up. Eight patients with traumatic high-flow priapism were reviewed. The patients were evaluated with clinical findings, laboratory examinations, and imaging findings including color Doppler ultrasonography and angiography. Diagnostic angiography demonstrated a connection between the cavernosal artery and the corpus cavernosum. Fistulas were embolized using autologous blood clot, polyvinyl alcohol particles, detachable coils, or acrylic glue. One or more procedures per patient were needed to achieve success. Eleven embolization procedures were performed in eight patients. Immediate resolution of priapism was obtained after the procedures. Three patients (37.5%) had recurrence of priapism in the subsequent 1-3 weeks and required a repeat procedure. After the final procedures, all patients had complete resolution of priapism. Normal recurrence of erectile function was obtained in six of the patients (75%) after the final embolization. Selective arterial embolization is a useful therapeutic option in the management of patients with high-flow priapism. Various materials can be used successfully as embolizing agents in the procedures according to the patient's status.


Hypothalamic expression and moonlight-independent changes of Cry3 and Per4 implicate their roles in lunar clock oscillators of the lunar-responsive Goldlined spinefoot.
Lunar cycle-associated physiology has been found in a wide variety of organisms. Studies suggest the presence of a circalunar clock in some animals, but the location of the lunar clock is unclear. We previously found lunar-associated expression of transcripts for Cryptochrome3 gene (SgCry3) in the brain of a lunar phase-responsive fish, the Goldlined spinefoot (Siganus guttatus). Then we proposed a photoperiodic model for the lunar phase response, in which SgCry3 might function as a phase-specific light response gene and/or an oscillatory factor in unidentified circalunar clock. In this study, we have developed an anti-SgCRY3 antibody to identify SgCRY3-immunoreactive cells in the brain. We found immunoreactions in the subependymal cells located in the mediobasal region of the diencephalon, a crucial site for photoperiodic seasonal responses in birds. For further assessment of the lunar-responding mechanism and the circalunar clock, we investigated mRNA levels of Cry3 as well as those of the other clock(-related) genes, Period (Per2 and Per4), in S. guttatus reared under nocturnal moonlight interruption or natural conditions. Not only SgCry3 but SgPer4 mRNA levels showed lunar phase-dependent variations in the diencephalon without depending on light condition during the night. These results suggest that the expressions of SgCry3 and SgPer4 are not directly regulated by moonlight stimulation but endogenously mediated in the brain, and implicate that circadian clock(-related) genes may be involved in the circalunar clock locating within the mediobasal region of the diencephalon.


Maternal protein intake during pregnancy and linear growth in the offspring.
Observational and experimental evidence demonstrates that protein intake in infancy programs linear growth. To our knowledge, few studies have examined prenatal maternal protein intake. Our objective was to examine associations of maternal protein intake during pregnancy with offspring linear growth. We analyzed data from 1961 mother-child pairs in Project Viva. We assessed first- and second-trimester diet with the use of food-frequency questionnaires and analyzed protein intake as grams per kilogram prepregnancy weight per day. We used research measures of offspring length at birth and in infancy (∼6 mo), early childhood (∼3 y), and midchildhood (∼7 y), as well as clinical growth measures obtained from after birth through midchildhood. We calculated sex-specific birth length z scores for gestational age with the use of international reference data. We used mixed models with repeated length measures to predict individual length gain velocities for birth to <6 mo and 6 mo to 7 y of age, then used these velocities as outcomes in adjusted linear regression models with maternal protein intake as the main predictor. Mean (range) second-trimester protein intake was 1.4 g · kg-1 · d-1 (0.3-3.1 g · kg-1 · d-1). After adjusting for maternal sociodemographics, gestational weight gain, maternal and paternal height, and (for postdelivery outcomes) child sex, gestational age, and breastfeeding duration, each 1-SD (0.36 g · kg-1 · d-1) increment in second-trimester protein intake corresponded to a -0.10 (95% CI: -0.18, -0.03) change in birth length z score, a -0.03 cm/mo (95% CI: -0.05, -0.01 cm/mo) change in slope of length growth from birth to <6mo, and a -0.09 cm/y (95% CI: -0.14, -0.05 cm/y) change in slope of length growth from 6 mo to midchildhood. Results were similar for first-trimester intake. In a population with relatively high protein intake during pregnancy, higher protein intake was associated with shorter offspring birth length and slower linear growth into midchildhood. Results suggest that higher protein intake during pregnancy does not increase fetal and child growth and may even reduce early length growth. Project Viva was registered at clinicaltrials.gov as NCT02820402.


Efficiency, behavior, and clinical properties of superelastic NiTi versus multistranded stainless steel wires: a prospective clinical trial.
To investigate efficiency, behavior, and properties of superelastic NiTi vs multistranded stainless steel wires in Begg and preadjusted edgewise appliance (PEA) under moderate to severe crowding conditions. Ninety-six participants (48 male, 48 female), aged 12-18 years old (mean age = 15.2 ± 1.95), with moderate (≤ 6 mm; mean = 5.3 ± 0.48) to severe (> 6 mm; mean = 7.9 ± 0.66) initial crowding were distributed into four groups: superelastic NiTi PEA (n = 24), superelastic NiTi Begg (n = 24), multistranded (coaxial) stainless steel PEA (n = 25), and multistranded (coaxial) stainless steel Begg (n = 23). In this study, 0.16-inch superelastic (austenitic active) NiTi and 0.175-inch multistranded (six stranded, coaxial) stainless steel wires were used in a 0.022-inch slot (Roth prescription) PEA and Begg appliance with a follow-up of six weeks. Analysis of variance revealed no significant difference in reduction of crowding between superelastic NiTi PEA and multistranded (coaxial) stainless steel PEA groups, but reduction in crowding was significantly greater in the superelastic NiTi Begg group compared with the multistranded (coaxial) stainless steel Begg group with F (3, 44) = 8.896, P < .001, and effect size (ω) 0.57 in moderate crowding and F (3, 44) = 122.341, P < .001, and effect size (ω) 0.93 in severe crowding. Linear regression demonstrated significant (P < .05) positive correlation between amount of initial crowding and reduction in crowding in all groups except the multistranded (coaxial) stainless steel Begg group, wherein a negative correlation did exist. Superelastic NiTi performed significantly better than multistranded (coaxial) stainless steel wire in the Begg appliance. However, in PEA, there was no significant difference.


Translation-linked mRNA destabilization accompanying serum-induced Nox4 expression in human endothelial cells.
NADPH oxidase is involved in cell signaling, regulating proliferation of vascular cells, especially in endothelium. The Nox4 catalytic subunit has a major role in endothelial cells, but growth arrest of cultured endothelial cells following serum deprivation paradoxically increases mRNA for Nox4. We investigated the relationships between Nox4 mRNA stability and protein expression in human microvascular endothelial cells. Serum starvation increased the steady-state level of Nox4 mRNA but paradoxically diminished Nox4 protein expression. mRNA transcription in the absence of serum is maintained by the p38MAP kinase pathway, for inhibition of p38MAP kinase reduced both Nox4 mRNA and Nox4 promoter activity. In serum-starved cells, reintroduction of serum increased Nox4 protein levels within 30 min and up to 24 h. In contrast, the mRNA decreased equally rapidly after serum stimulation. Inhibition of Nox4 translation by cycloheximide blocked serum-induced mRNA degradation and Nox4 protein synthesis, and actinomycin-D also delayed Nox4 mRNA decay. Therefore, Nox4 mRNA level falls after serum stimulation because of a translation-initiated mRNA destabilization program. Clearly Nox4 mRNA is regulated at both transcriptional and post-transcriptional levels, and the steady state level of Nox4 mRNA does not accurately reflect Nox4 protein abundance and functions, with implications for regulation of cell proliferation and survival.


The vertebrate Hef ortholog is a component of the Fanconi anemia tumor-suppressor pathway.
The helicase-associated endonuclease for fork-structured DNA (Hef) is an archaeabacterial protein that processes blocked replication forks. Here we have isolated the vertebrate Hef ortholog and investigated its molecular function. Disruption of this gene in chicken DT40 cells results in genomic instability and sensitivity to DNA cross-links. The similarity of this phenotype to that of cells lacking the Fanconi anemia-related (FA) tumor-suppressor genes led us to investigate whether Hef functions in this pathway. Indeed, we found a genetic interaction between the FANCC and Hef genes. In addition, Hef is a component of the FA nuclear protein complex that facilitates its DNA damage-inducible chromatin localization and the monoubiquitination of the FA protein FANCD2. Notably, Hef interacts directly with DNA structures that are intermediates in DNA replication. This discovery sheds light on the origins, regulation and molecular function of the FA tumor-suppressor pathway in the maintenance of genome stability.


Probing resonating valence bond states in artificial quantum magnets.
Designing and characterizing the many-body behaviors of quantum materials represents a prominent challenge for understanding strongly correlated physics and quantum information processing. We constructed artificial quantum magnets on a surface by using spin-1/2 atoms in a scanning tunneling microscope (STM). These coupled spins feature strong quantum fluctuations due to antiferromagnetic exchange interactions between neighboring atoms. To characterize the resulting collective magnetic states and their energy levels, we performed electron spin resonance on individual atoms within each quantum magnet. This gives atomic-scale access to properties of the exotic quantum many-body states, such as a finite-size realization of a resonating valence bond state. The tunable atomic-scale magnetic field from the STM tip allows us to further characterize and engineer the quantum states. These results open a new avenue to designing and exploring quantum magnets at the atomic scale for applications in spintronics and quantum simulations.


Perseveration related to frontal lesion in mice using the olfactory H-maze.
The delayed reaction paradigm, consisting to discover two different rules consecutively (delayed alternation and non-alternation task) followed by a delayed reversal task, is a specific marker for the functioning of primate prefrontal cortex. Although several works in rodents report the use of operant delayed alternation tasks, in none of the studies mice with lesion of the prefrontal cortex were used in this paradigm. In the current study, mouse experiments were conducted using a new, totally automated device, the olfactory H-maze. Here, we show that unilateral lesion of the dorsomedial prefrontal cortex in mice induced similar deficits to those observed after frontal lesions in monkeys and humans. These pronounced learning deficits seem to come from difficulty elaborating a new rule and the inability to inhibit the previous rule, characterized by perseveration after prefrontal cortex lesion. The present results demonstrate that this very simple experimental paradigm using the olfactory H-maze presents the advantage to be fast (one training session) and well suited to assess the frontal functions in mice. It should be useful for testing pharmacological or stem cell approaches in order to reduce organic damages or gain insight into the cognitive functions of the frontal cortex using transgenic or gene-targeting mice.


Adaptation and Implementation of a Family Caregiver Skills Training Program: From Single Site RCT to Multisite Pragmatic Intervention.
We describe an approach to rapidly adapt and implement an education and skills improvement intervention to address the needs of family caregivers of functionally impaired veterans-Helping Invested Families Improve Veterans' Experience Study (HI-FIVES). Prior to implementation in eight sites, a multidisciplinary study team made systematic adaptations to the curriculum content and delivery process using input from the original randomized controlled trial (RCT); a stakeholder advisory board comprised of national experts in caregiver education, nursing, and implementation; and a veteran/caregiver engagement panel. To address site-specific implementation barriers in diverse settings, we applied the Replicating Effective Programs implementation framework. Adaptations to HI-FIVES content and delivery included identifying core/noncore curriculum components, reducing instruction time, and simplifying caregiver recruitment for clinical settings. To enhance curriculum flexibility and potential uptake, site personnel were able to choose which staff would deliver the intervention and whether to offer class sessions in person or remotely. Curriculum materials were standardized and packaged to reduce the time required for implementation and to promote fidelity to the intervention. The emphasis on flexible intervention delivery and standardized materials has been identified as strengths of the adaptation process. Two key challenges have been identifying feasible impact measures and reaching eligible caregivers for intervention recruitment. This systematic implementation process can be used to rapidly adapt an intervention to diverse clinical sites and contexts. Nursing professionals play a significant role in educating and supporting caregivers and care recipients and can take a leading role to implement interventions that address skills and unmet needs for caregivers.


Drop friction on liquid-infused materials.
We discuss in this paper the nature of the friction generated as a drop glides on a textured material infused by another liquid. Different regimes are found, depending on the viscosities of both liquids. While a viscous drop simply obeys a Stokes-type friction, the force opposing a drop moving on a viscous substrate becomes non-linear in velocity. A liquid on an infused material is surrounded by a meniscus, and this specific feature is proposed to be responsible for the special frictions observed on both adhesive and non-adhesive substrates.


"Medically unexplained" symptoms and symptom disorders in primary care: prognosis-based recognition and classification.
Many patients consult their GP because they experience bodily symptoms. In a substantial proportion of cases, the clinical picture does not meet the existing diagnostic criteria for diseases or disorders. This may be because symptoms are recent and evolving or because symptoms are persistent but, either by their character or the negative results of clinical investigation cannot be attributed to disease: so-called "medically unexplained symptoms" (MUS). MUS are inconsistently recognised, diagnosed and managed in primary care. The specialist classification systems for MUS pose several problems in a primary care setting. The systems generally require great certainty about presence or absence of physical disease, they tend to be mind-body dualistic, and they view symptoms from a narrow specialty determined perspective. We need a new classification of MUS in primary care; a classification that better supports clinical decision-making, creates clearer communication and provides scientific underpinning of research to ensure effective interventions. We propose a classification of symptoms that places greater emphasis on prognostic factors. Prognosis-based classification aims to categorise the patient's risk of ongoing symptoms, complications, increased healthcare use or disability because of the symptoms. Current evidence suggests several factors which may be used: symptom characteristics such as: number, multi-system pattern, frequency, severity. Other factors are: concurrent mental disorders, psychological features and demographic data. We discuss how these characteristics may be used to classify symptoms into three groups: self-limiting symptoms, recurrent and persistent symptoms, and symptom disorders. The middle group is especially relevant in primary care; as these patients generally have reduced quality of life but often go unrecognised and are at risk of iatrogenic harm. The presented characteristics do not contain immediately obvious cut-points, and the assessment of prognosis depends on a combination of several factors. Three criteria (multiple symptoms, multiple systems, multiple times) may support the classification into good, intermediate and poor prognosis when dealing with symptoms in primary care. The proposed new classification specifically targets the patient population in primary care and may provide a rational framework for decision-making in clinical practice and for epidemiologic and clinical research of symptoms.


Use of newer antiretroviral treatments among HIV-infected medicaid beneficiaries with serious mental illness.
The study compares rates of protease inhibitor (PI) use during the 3 years following the introduction of these newer treatments among human immunodeficiency virus (HIV)-infected individuals with and without serious mental illness and examines persistence of use of these therapies across these subgroups. We used merged autoimmune deficiency syndrome (AIDS)/HIV surveillance and Medicaid claims data to examine use of PIs and non-nucleoside reverse transcriptase inhibitors (NNRTIs) among New Jersey Medicaid beneficiaries with AIDS between 1996 and 1998. Based on the ICD-9-CM diagnoses assigned by a high-credibility source in 1 inpatient or 2 outpatient claims, we identified patients with schizophrenia (ICD-9-CM code 295) and those with severe affective disorder (combining patients with recurrent major depressive disorder [ICD-9-CM code 296.3] or bipolar disorder [296.4, 296.5, 296.6, 296.7, or 296.8]). These groups were compared with those patients with no serious mental illness. In this sample, patients with schizophrenia (68.3%) and those with severe affective disorder (75.6%) were more likely to have initiated new antiretroviral therapy than were those without serious mental illness (64.3%). Patients with severe affective disorder, but not those with schizophrenia, were significantly less persistent (p <.01) in their use of PI/NNRTI therapy than those without serious mental illness. No evidence was found that the presence of a serious mental illness discourages physicians from initiating new antiretroviral therapy, perhaps reflecting a comparatively high level of integration of these patients into the health care system. Patients with schizophrenia are as persistent in their use of PI/NNRTI therapy as those without a serious mental illness. Lower rates of medication compliance by those with severe affective disorder justify increased efforts to support optimal adherence.


Analysis of drug coverage before and after the implementation of Canada's Common Drug Review.
Canada's Common Drug Review was implemented to provide publicly funded drug plans (provincial and federal) with a transparent, rigorous and consistent approach for assessing the clinical effectiveness and cost-effectiveness of new drugs. We compared uptake of drug coverage across jurisdictions before and after the implementation of the Common Drug Review. Using the IMS Brogan formulary acceptance: monitoring and evaluation database, we identified new drug products in Canada five years before and five years after the first recommendation was made by the Common Drug Review. For each jurisdiction, we compared the proportion of drugs listed, the median time-to-listing and the agreement between the listing decisions of the drug plans and the recommendations of the Common Drug Review. We identified 198 new drugs approved for use in Canada between May 26, 1999, and May 27, 2009, of which 53 had a recommendation from the Common Drug Review. The proportion of drugs listed decreased after the introduction of the Common Drug Review for all participating drug plans (81.1% to 71.3% overall [p ≤ 0.01 for all plans, with the exceptions of Ontario and Quebec [p = 0.07]). The change in median time-to-listing between the periods before and after the Common Drug Review varied by jurisdiction, ranging from a decrease of 691 days to an increase of 250 days. The change in median time-to-listing was not statistically significant for most jurisdictions, with the exceptions of Saskatchewan (increased, Mann-Whitney U test p = 0.01) and New Brunswick, Prince Edward Island, and Newfoundland and Labrador (all decreased, Mann-Whitney U test p < 0.01). There was a decline in the proportion of new drugs listed after the introduction of the Common Drug Review for both participating and nonparticipating jurisdictions. The introduction of the review was associated with a decreased time-to-listing for certain smaller provinces.


Informing the UK Muslim Community on Organ Donation: Evaluating the Effect of a National Public Health Programme by Health Professionals and Faith Leaders.
There is a significant shortage of transplantable organs in the UK particularly from Black, Asian and Minority Ethnic (BAME) groups, of which Muslims make a large proportion. The British Islamic Medical Association (BIMA) held a nationwide series of community gatherings with the aim of describing the beliefs and attitudes to organ donation amongst British Muslims and evaluate the efficacy of a national public health programme on views and uncertainties regarding religious permissibility and willingness to register. Eight public forums were held across the UK between June 2019 and March 2020 by the British Islamic Medical Association (BIMA). A panel of experts consisting of health professionals and Imams discussed with audiences the procedures, experiences and Islamic ethico-legal rulings on organ donation. Attendees completed a self-administered questionnaire which captured demographic data along with opinions before and after the session regarding religious permissibility and willingness to register given permissibility. A total of 554 respondents across seven UK cities were included with a M:F ratio 1:1.1. Only 45 (8%) respondents were registered as organ donors. Amongst those not registered multiple justifications were detailed, foremost of which was religious uncertainty (73%). Pre-intervention results indicated 50% of respondents were unsure of the permissibility of organ donation in Islam. Of those initially unsure or against permissibility or willingness to register, 72% changed their opinion towards deeming it permissible and 60% towards a willingness to register indicating a significant change in opinion (p < 0.001). The effectiveness of our interventions suggests further education incorporating faith leaders alongside local healthcare professionals to address religious and cultural concerns can reduce uncertainty whilst improving organ donation rates among the Muslim community.


Outpacing the pneumococcus: Antibody dynamics in the first few days following pneumococcal capsular antigen stimulation.
Children in developing countries are frequently exposed to the pneumococcus, but few develop invasive pneumococcal disease (IPD). We test the hypothesis that natural variation exists in the rapidity of IgG responses following exposure to pneumococcal polysaccharides, and that these differences are sufficiently great to affect susceptibility to and outcome of IPD. We recruited children aged 24-36 months, who had recovered from IPD, and age-matched healthy controls and vaccinated them with 1 dose of the 23-valent PPV to mimic natural exposure. We collected serum samples after vaccination and analysed the dynamics of anti-polysaccharide antibody responses to several capsular antigens. Mean IgG response times to different serotypes were 6.4-7.3 days, with standard deviations of 0.9-1.85 days, suggesting a natural range in response times of up to 7 days. Serotype 1 elicited the largest fold-rise, serotype 23F the smallest. The proportion of responses achieved by day 7 was similar in children with a history of IPD and healthy children. There was considerable natural variation in the rapidity of anti-capsular IgG responses extending over 4-7 days. There was no evidence to suggest that children who have experienced IPD respond more slowly to heterologous pneumococcal capsular antigens than do healthy children.


Urbanization and blood parasite infections affect the body condition of wild birds.
Human landscape transformation, especially urbanization, strongly affects ecosystems worldwide. Both urban stressors and parasites have negative effects on organism health, however the potential synergy between those factors has been poorly investigated. We analysed the body condition (i.e. body mass after controlling for wing chord) of 2043 house sparrows (adults and yearlings) captured in 45 localities along an urbanization gradient in relation to Plasmodium, Haemoproteus and Leucocytozoon infection status. Body condition was negatively related to urbanization level and to urbanized land coverage but only in yearling birds from urban habitats. In addition, bird body condition tended to increase in rural habitats, significantly in the case of yearlings. Infected individuals by Plasmodium or Haemoproteus had higher body condition than un-infected birds, but this pattern could be due to a selective disappearance of infected individuals with lower body condition as suggested by the reduced variance in body condition in infected birds in urban habitats. These results provide support for a negative impact of urbanization on bird body condition, while Plasmodium and Haemoproteus may exert selection against individuals with lower body condition living in urban habitats, especially during earlier life stages, underlining the synergistic effects that urbanization and parasites may have on wild birds.


Constituents of Music and Visual-Art Related Pleasure - A Critical Integrative Literature Review.
The present literature review investigated how pleasure induced by music and visual-art has been conceptually understood in empirical research over the past 20 years. After an initial selection of abstracts from seven databases (keywords: pleasure, reward, enjoyment, and hedonic), twenty music and eleven visual-art papers were systematically compared. The following questions were addressed: (1) What is the role of the keyword in the research question? (2) Is pleasure considered a result of variation in the perceiver's internal or external attributes? (3) What are the most commonly employed methods and main variables in empirical settings? Based on these questions, our critical integrative analysis aimed to identify which themes and processes emerged as key features for conceptualizing art-induced pleasure. The results demonstrated great variance in how pleasure has been approached: In the music studies pleasure was often a clear object of investigation, whereas in the visual-art studies the term was often embedded into the context of an aesthetic experience, or used otherwise in a descriptive, indirect sense. Music studies often targeted different emotions, their intensity or anhedonia. Biographical and background variables and personality traits of the perceiver were often measured. Next to behavioral methods, a common method was brain imaging which often targeted the reward circuitry of the brain in response to music. Visual-art pleasure was also frequently addressed using brain imaging methods, but the research focused on sensory cortices rather than the reward circuit alone. Compared with music research, visual-art research investigated more frequently pleasure in relation to conscious, cognitive processing, where the variations of stimulus features and the changing of viewing modes were regarded as explanatory factors of the derived experience. Despite valence being frequently applied in both domains, we conclude, that in empirical music research pleasure seems to be part of core affect and hedonic tone modulated by stable personality variables, whereas in visual-art research pleasure is a result of the so called conceptual act depending on a chosen strategy to approach art. We encourage an integration of music and visual-art into to a multi-modal framework to promote a more versatile understanding of pleasure in response to aesthetic artifacts.


A metal-organic framework based nanocomposite with co-encapsulation of Pd@Au nanoparticles and doxorubicin for pH- and NIR-triggered synergistic chemo-photothermal treatment of cancer cells.
Here, we report a novel metal-organic framework-based nanocomposite with encapsulated Pd@Au nanoparticles and doxorubicin (DOX) for pH- and NIR-triggered synergistic chemo-photothermal treatment of cancer cells. In this work, Pd nanoparticles, which have uniform size and dispersibility, were first synthesized and used as a template to direct the covering of Au nanosheets. The obtained Au coated Pd (Pd@Au) nanoparticles have excellent dispersibility and photothermal conversion ability, which makes them a good photothermal nanomaterial. Subsequently, an acid-degradable metal-organic framework of ZIF-8 was employed to synchronously encapsulate Pd@Au nanoparticles and DOX to get a metal-organic framework-based nanocomposite (DOX/Pd@Au@ZIF-8). Under acid conditions (e.g. pH ∼5.0 in a lysosome), the ZIF-8 framework of the DOX/Pd@Au@ZIF-8 nanocomposite could be degraded, resulting in the release of encapsulated DOX. Moreover, the present Pd@Au nanoparticles can effectively convert NIR laser light (780 nm, 2.1 W cm-2) into heat, not only further promoting the release of DOX, but also realizing the synergistic chemo-photothermal treatment of cancer cells. The in vitro experiments showed that this nanocomposite system has an excellent synergistic treatment effect on SMMC-7721 cells, even at low concentrations (e.g. 20 μg mL-1). With the properties of synergistic chemo-photothermal treatment, we hope that such a nanocomposite system of DOX/Pd@Au@ZIF-8 could open the door to designing a significant multifunctional system for diverse applications in cancer treatment.


Inadequate Iodine Intake in Population Groups Defined by Age, Life Stage and Vegetarian Dietary Practice in a Norwegian Convenience Sample.
Inadequate iodine intake has been identified in populations considered iodine replete for decades. The objective of the current study is to evaluate urinary iodine concentration (UIC) and the probability of adequate iodine intake in subgroups of the Norwegian population defined by age, life stage and vegetarian dietary practice. In a cross-sectional survey, we assessed the probability of adequate iodine intake by two 24-h food diaries and UIC from two fasting morning spot urine samples in 276 participants. The participants included children (n = 47), adolescents (n = 46), adults (n = 71), the elderly (n = 23), pregnant women (n = 45), ovo-lacto vegetarians (n = 25), and vegans (n = 19). In all participants combined, the median (95% CI) UIC was 101 (90, 110) µg/L, median (25th, 75th percentile) calculated iodine intake was 112 (77, 175) µg/day and median (25th, 75th percentile) estimated usual iodine intake was 101 (75, 150) µg/day. According to WHOs criteria for evaluation of median UIC, iodine intake was inadequate in the elderly, pregnant women, vegans and non-pregnant women of childbearing age. Children had the highest (82%) and vegans the lowest (14%) probability of adequate iodine intake according to reported food and supplement intakes. This study confirms the need for monitoring iodine intake and status in nationally representative study samples in Norway.


Effect of airway impedance on CO2 retention and respiratory muscle activity during NREM sleep.
We hypothesized that a sleep-induced increase in mechanical impedance contributes to CO2 retention and respiratory muscle recruitment during non-rapid-eye-movement (NREM) sleep. The effect NREM sleep on respiratory muscle activity and CO2 retention was measured in healthy subjects who increased maximum total pulmonary resistance (RLmax, 1-81 cmH2O.l-1.s) from awake to NREM sleep. We determined the effects of this sleep-induced increase in airway impedance by steady-state inhalation of a reduced-density gas mixture (79% He-21% O2, He-O2). Both arterialized blood PCO2 (PaCO2) and end-tidal PCO2 (PETCO2) were measured. Inspiratory (EMGinsp) and expiratory (EMGexp) respiratory muscle electromyogram activity was measured. NREM sleep caused 1) RLmax to increase (7 +/- 3 vs. 39 +/- 28 cmH2O.l-1.s), 2) PaCO2 and/or PETCO2 to increase in all subjects (40 +/- 2 vs. 44 +/- 3 Torr), and 3) EMGinsp to increase in 8 of 9 subjects and EMGexp to increase in 9 of 17 subjects. Compared with steady-state air breathing during NREM sleep, steady-state He-O2 breathing 1) reduced RLmax by 38%, 2) decreased PaCO2 and PETCO2 by 2 Torr, and 3) decreased both EMGinsp (-20%) and EMGexp (-54%). We concluded that the sleep-induced increase in upper airway resistance accompanied by the absence of immediate load compensation is an important determinant of CO2 retention, which, in turn, may cause augmentation of inspiratory and expiratory muscle activity above waking levels during NREM sleep.


Fast and accurate dose predictions for novel radiotherapy treatments in heterogeneous phantoms using conditional 3D-UNet generative adversarial networks.
Novel radiotherapy techniques like synchrotron X-ray microbeam radiation therapy (MRT) require fast dose distribution predictions that are accurate at the sub-mm level, especially close to tissue/bone/air interfaces. Monte Carlo (MC) physics simulations are recognized to be one of the most accurate tools to predict the dose delivered in a target tissue but can be very time consuming and therefore prohibitive for treatment planning. Faster dose prediction algorithms are usually developed for clinically deployed treatments only. In this work, we explore a new approach for fast and accurate dose estimations suitable for novel treatments using digital phantoms used in preclinical development and modern machine learning techniques. We develop a generative adversarial network (GAN) model, which is able to emulate the equivalent Geant4 MC simulation with adequate accuracy and use it to predict the radiation dose delivered by a broad synchrotron beam to various phantoms. The energy depositions used for the training of the GAN are obtained using full Geant4 MC simulations of a synchrotron radiation broad beam passing through the phantoms. The energy deposition is scored and predicted in voxel matrices of size 140 × 18 × 18 with a voxel edge length of 1 mm. The GAN model consists of two competing 3D convolutional neural networks, which are conditioned on the photon beam and phantom properties. The generator network has a U-Net structure and is designed to predict the energy depositions of the photon beam inside three phantoms of variable geometry with increasing complexity. The critic network is a relatively simple convolutional network, which is trained to distinguish energy depositions predicted by the generator from the ones obtained with the full MC simulation. The energy deposition predictions inside all phantom geometries under investigation show deviations of less than 3% of the maximum deposited energy from the simulation for roughly 99% of the voxels in the field of the beam. Inside the most realistic phantom, a simple pediatric head, the model predictions deviate by less than 1% of the maximal energy deposition from the simulations in more than 96% of the in-field voxels. For all three phantoms, the model generalizes the energy deposition predictions well to phantom geometries, which have not been used for training the model but are interpolations of the training data in multiple dimensions. The computing time for a single prediction is reduced from several hundred hours using Geant4 simulation to less than a second using the GAN model. The proposed GAN model predicts dose distributions inside unknown phantoms with only small deviations from the full MC simulation with computations times of less than a second. It demonstrates good interpolation ability to unseen but similar phantom geometries and is flexible enough to be trained on data with different radiation scenarios without the need for optimization of the model parameter. This proof-of-concept encourages to apply and further develop the model for the use in MRT treatment planning, which requires fast and accurate predictions with sub-mm resolutions.


Clinical case reports and case series research in evaluating surgery. Part II. The content and form: uses of single clinical case reports and case series research in surgical specialties.
Single clinical case and case series reports have long been an integral part of medicine and surgery. Despite this, due to their vague objectives and questions to answer, unstructured presentations and absent, exaggerated and/or unsubstantiated conclusions and recommendations, they are sometimes perceived as a lighter piece of evidence in modern surgical understanding and decision-making. Their value may, in fact, be in their contribution to theory, inference, or both. Today, modern case reporting is a balanced mixture of traditional patient-centered reporting methodology, a quantitative approach to ("scientific") inference and growing experience from qualitative research and evaluation methodology in general policies, programs, politics, arts, humanities, business, economics, and the military. Integration of such experiences is possible and even desirable in modern clinical case(s) reporting. Both strengths and weaknesses of case(s) reporting are based on the structure and content of case(s) reports in terms of the clarity of their leading questions and objectives, structured presentation in form and valid argument building blocks founded on the best evidence available leading to their realistic conclusions and claims. Readers must assess their real value in general and in the context of their own decisions in research and practice with patients under their care and in decision-making. Most discussion about single clinical case reports and case series research revolves around their relevance to some degree of proof of cause-effect relationships. The relevance of case reports and case series research goes well beyond this point. Recommendations for both surgical and other case presenters and their interlocutors and readers are offered for consideration.


Splice variants of the nuclear dot-associated Sp100 protein contain homologies to HMG-1 and a human nuclear phosphoprotein-box motif.
Sp100 and PML are interferon-inducible proteins associated with a new class of nuclear domains (known as nuclear dots or PML bodies) which play a role in tumorigenesis, virus infections, and autoimmunity. While PML is extensively alternatively spliced, only two splice variants are known for Sp100. Here we describe the identification and characterization of several Sp100 splice variant proteins and support their existence by elucidation of the 3'-end of the Sp100 gene. Some of the splice variants contain a domain of significant sequence similarity with two previously described highly related interferon-inducible nuclear phosphoproteins as well as to suppressin and DEAF-1, which altogether define a novel protein motif, termed HNPP-box. One class of splice variants contains an almost complete and highly conserved copy of the DNA-binding high mobility group 1 protein sequence and thus represent novel HMG-box proteins. When expressed transiently, both major classes of Sp100 splice variant proteins localize in part to nuclear dots/PML bodies and in addition to different nuclear domains. Furthermore, PML was occasionally redistributed. These data indicate that alternatively spliced Sp100 proteins are expressed, differ in part in localization from Sp100, and might bind to chromatin via the HMG domain.


Medication and skilled work.
There is increasing interest in the way in which drugs impair performance. This has arisen because some may impair day-to-day skills of those whose occupations demand vigilance and motor skill, and of those who are involved in decision making or where interpersonal relations are crucial. For many years the position was adopted, at least in certain occupations where impaired performance could be a danger to others, that the use of any drug should preclude employment. However, recent advances in therapeutics and a greater understanding of drug action in man has made this rather uncomplicated view of life less tenable, and there is now an increasing desire that advances in therapy should, if at all possible, be available to occupational groups, such as airline pilots. In this way the adverse effect which a drug may have on performance has become an important aspect of its clinical profile. Hypnotics appropriate for transient insomnia, which may arise from the irregularity of rest inherent in many occupations, need to be free of residual effects, antihistamines that are sedative must be avoided, and drugs used in the management of mild hypertension, often during the important years of middle life, must be as free as possible from central effects. And it must be emphasized that these drugs are often used by active, healthy or near healthy individuals. The issues involved in the safe use of a particular drug by a particular individual are complex, and as with all aspects of therapeutics it is sometimes necessary to balance efficacy and adverse effects.


Differential metabolic activity by dental plaque bacteria in association with two preparations of MUC5B mucins in solution and in biofilms.
Salivary mucin, MUC5B, is an oligomeric glycoprotein, heterogeneous in size and with a diverse repertoire of oligosaccharides, which differ in composition and charge. Since complex salivary glycoproteins are considered to be the major source of nutrients for the oral supragingival microbiota, the major aim of the current study was to determine whether different preparations of non-denatured MUC5B could be isolated exhibiting different biological properties in relation to the microflora associated with the surfaces of the oral cavity. Two preparations, solMUC5B and gelMUC5B, were isolated by density-gradient centrifugation and were shown to have different buoyant densities, carbohydrate content and surface-adsorbing characteristics. To ascertain differences in biological activity, the two mucin preparations, both in solution and adsorbed to a model surface, were incubated with freshly isolated dental plaque and assayed for metabolic (dehydrogenase) activity with the fluoresecent substrate CTC (5-cyano-2,3-ditolyl tetrazolium chloride). The plaque bacteria exhibited higher metabolism with the solMUC5B preparation in solution, with 79.4 % active plaque cells compared to the controls without mucin (9.6 %), while gelMUC5B showed 48.2 % active cells with the same plaque population. In contrast, the same mucins adhered to a surface elicited a significantly lower metabolic response, with surface-associated plaque cells showing only 12.1 % active cells with solMUC5B and 29.2 % with gelMUC5B. These results suggested that the metabolism by the plaque cells adsorbed to surface-associated mucins was downregulated compared to the same cells suspended in mucin solution. This was confirmed in an experiment where active dispersed plaque/solMUC5B suspensions were shown to lose significant metabolic activity (e.g. 74.9 to 19.3 %) when allowed to interact with gelMUC5B adsorbed to a surface. Clearly, the solMUC5B and gelMUC5B preparations exhibited different biological activity when assayed with freshly plaque bacteria in suspension and in a biofilm.


Load-dependent assembly of the bacterial flagellar motor.
It is becoming clear that the bacterial flagellar motor output is important not only for bacterial locomotion but also for mediating the transition from liquid to surface living. The output of the flagellar motor changes with the mechanical load placed on it by the external environment: at a higher load, the motor runs more slowly and produces higher torque. Here we show that the number of torque-generating units bound to the flagellar motor also depends on the external mechanical load, with fewer stators at lower loads. Stalled motors contained at least as many stators as rotating motors at high load, indicating that rotation is unnecessary for stator binding. Mutant stators incapable of generating torque could not be detected around the motor. We speculate that a component of the bacterial flagellar motor senses external load and mediates the strength of stator binding to the rest of the motor. The transition between liquid living and surface living is important in the life cycles of many bacteria. In this paper, we describe how the flagellar motor, used by bacteria for locomotion through liquid media and across solid surfaces, is capable of adjusting the number of bound stator units to better suit the external load conditions. By stalling motors using external magnetic fields, we also show that rotation is not required for maintenance of stators around the motor; instead, torque production is the essential factor for motor stability. These new results, in addition to previous data, lead us to hypothesize that the motor stators function as mechanosensors as well as functioning as torque-generating units.


The p66shc adaptor protein controls oxidative stress response and life span in mammals.
Gene mutations in invertebrates have been identified that extend life span and enhance resistance to environmental stresses such as ultraviolet light or reactive oxygen species. In mammals, the mechanisms that regulate stress response are poorly understood and no genes are known to increase individual life span. Here we report that targeted mutation of the mouse p66shc gene induces stress resistance and prolongs life span. p66shc is a splice variant of p52shc/p46shc (ref. 2), a cytoplasmic signal transducer involved in the transmission of mitogenic signals from activated receptors to Ras. We show that: (1) p66shc is serine phosphorylated upon treatment with hydrogen peroxide (H2O2) or irradiation with ultraviolet light; (2) ablation of p66shc enhances cellular resistance to apoptosis induced by H2O2 or ultraviolet light; (3) a serine-phosphorylation defective mutant of p66shc cannot restore the normal stress response in p66shc-/- cells; (4) the p53 and p21 stress response is impaired in p66shc-/- cells; (5) p66shc-/- mice have increased resistance to paraquat and a 30% increase in life span. We propose that p66shc is part of a signal transduction pathway that regulates stress apoptotic responses and life span in mammals.


The emergence of 'omics for the management of colorectal cancer.
The past couple of years have seen a flurry of publications detailing potential prognostic and predictive biomarkers for colorectal cancer. However, most studies have been underpowered and demonstrate elements of a statistical 'fishing exercise', that is the carrying out of a huge number of analyses to find a hint of importance for a particular marker. Recently some well powered analyses using separate development and validation datasets and multivariate statistics have been published and these are starting to change attitudes towards such markers. Although there are very limited markers in standard clinical practice, we feel that there are now some that are ready to be translated into routine use or that at least deserve some further attention and 'service assessment'.


Hydrogen-terminated diamond electrodes. I. Charges, potentials, and energies.
Hydrogen-terminated intrinsic diamond is a most unusual insulator since in contact with air or electrolyte it develops a hole accumulation layer just below the surface. When immersed in electrolyte the hole concentration responds to ionic charges and potentials and this response can be monitored by measuring the conductance of the hole accumulation layer without interference form the bulk conductivity. This feature has been widely used for chemical sensors in the form of solution-gate field effect transistors (SGFET). Here we analyze the charge and the potential profiles in the diamond and the electrolyte, as well as the static differential capacitance of the diamond electrode under controlled potential conditions. From this analysis we derive expressions for the transfer characteristics of diamond-based SGFET's that faithfully describe experimental data presented here as well. This holds in particular for the threshold region of the transfer characteristics that can only be modeled if the unusual semiconducting properties of the diamond electrode are taken into account properly. From fits to our data we derive (among other things) a value of chi=-0.50+/-0.02eV for the electron affinity of the hydrogen-terminated diamond surface in contact with aqueous electrolyte.


Infection of peritoneal macrophages in vitro and in vivo with feline immunodeficiency virus.
Macrophages were harvested from the peritoneal cavities of healthy specific-pathogen-free cats by saline lavage. Three days before collection, the peritoneal cavities were stimulated with glutaraldehyde-fixed Saccharomyces cerevisiae cells to induce greater numbers of macrophages and to begin the activation sequence. Peritoneal macrophages from cats stimulated once with yeast consisted mainly of small macrophages and a smaller number of larger activated macrophages. After several days in culture, many of the small macrophages became activated and a portion of the activated macrophages developed into multinucleated giant cells. Peritoneal cells from cats that were stimulated twice or three times with yeast at 3-week intervals consisted of a higher proportion of activated macrophages initially and produced more and larger multinuclear giant cells with time. Cultures of peritoneal cells stimulated once with yeast were easily infected in vitro with feline immunodeficiency virus (FIV) and produced a transient burst of reverse transcriptase activity. After the initial burst of virus replication, the infection became latent. Even more multinucleated giant cells appeared after infection, and many of these cells fused with each other. Replicating virus could be rescued from the latently infected macrophages after 2 to 3 weeks of phorbol myristate acetate stimulation and cocultivation with T-lymphocyte-enriched peripheral blood mononuclear cells. Multiply stimulated peritoneal cells, which contained a much higher proportion of activated macrophages, could also be infected in vitro with FIV. The infection usually became latent, however, without going through an initial replicative stage. Peritoneal cells from chronically FIV-infected specific-pathogen-free cats contained a higher proportion of activated macrophages and were latently infected with FIV from the outset.


Monocyte maturation, HIV susceptibility, and transmigration across the blood brain barrier are critical in HIV neuropathogenesis.
HIV continues to be a global health crisis with more than 34 million people infected worldwide (UNAIDS: Report on the Global AIDS Epidemic 2010, Geneva, World Health Organization). HIV enters the CNS within 2 weeks of infection and establishes a spectrum of HAND in a large percentage of infected individuals. These neurologic deficits greatly impact the quality of life of those infected with HIV. The establishment of HAND is largely attributed to monocyte transmigration, particularly that of a mature CD14(+)CD16(+) monocyte population, which is more susceptible to HIV infection, across the BBB into the CNS parenchyma in response to chemotactic signals. To enter the CNS, junctional proteins on the monocytes must participate in homo- and heterotypic interactions with those present on BMVECs of the BBB as they transmigrate across the barrier. This transmigration is responsible for bringing virus into the brain and establishing chronic neuroinflammation. While there is baseline trafficking of monocytes into the CNS, the increased chemotactic signals present during HIV infection of the brain promote exuberant monocyte transmigration into the CNS. This review will discuss the mechanisms of monocyte differentiation/maturation, HIV infectivity, and transmigration into the CNS parenchyma that contribute to the establishment of cognitive impairment in HIV-infected individuals. It will focus on markers of monocyte subpopulations, how differentiation/maturation alters HIV infectivity, and the mechanisms that promote their increased transmigration across the BBB into the CNS.


The distribution and sources of polycyclic aromatic hydrocarbons in surface sediments along the Egyptian Mediterranean coast.
Coastal marine sediment samples were collected from 31 sampling stations along the Egyptian Mediterranean Sea coast. All sediment samples were analyzed to determine aliphatic and polycyclic aromatic hydrocarbons (PAHs) as well as total organic carbon (TOC) contents and grain size analysis. Total concentrations of 16 EPA-PAHs in the sediments were varied from 88 to 6338 ng g(-1) with an average value of 154 ng g(-1) (dry weight). However, the concentrations of total aliphatic were varied from 1.3 to 69.9 ng g(-1) with an average value of 15.6 ng g(-1) (dry weight). The highest contents of PAHs were found in the Eastern harbor (6338 ng g(-1)), Manzala (5206 ng g(-1)) and El-Jamil East (4895 ng g(-1)) locations. Good correlations observed between a certain numbers of PAH concentrations allowed to identify its origin. The average total organic carbon (TOC) percent was varied from 0.91 to 4.54%. Higher concentration of total pyrolytic hydrocarbons ( summation operatorCOMB) than total fossil hydrocarbons ( summation operatorPHE) declared that atmospheric fall-out is the significant source of PAHs to marine sediments of the Egyptian Mediterranean coast. The selected marked compounds, a principal component analysis (PCA) and special PAHs compound ratios (phenanthrene/anthracene vs fluoranthene/pyrene; summation operatorCOMB/ summation operatorEPA-PAHs) suggest the pyrogenic origins, especially traffic exhausts, are the dominant sources of PAHs in most locations. Interferences of rather petrogenic and pyrolytic PAH contaminations were noticed in the harbors due to petroleum products deliveries and fuel combustion emissions from the ships staying alongside the quays.


Serum galectin-9 as a noninvasive biomarker for the detection of endometriosis and pelvic pain or infertility-related gynecologic disorders.
To investigate the usefulness of soluble galectin-9 (Gal-9) in the noninvasive laboratory diagnosis of endometriosis and various gynecologic disorders. Prospective case-control study. University medical centers. A total of 135 women of reproductive age were involved in the study, 77 endometriosis patients, 28 gynecologic controls, and 30 healthy women. Diagnostic laparoscopy and collection of tissue biopsies, peritoneal cells, and native peripheral blood from different case groups of gynecology patients and healthy women. The expression of mRNA and serum concentration of Gal-9. Semiquantitative reverse transcription-polymerase chain reaction analysis and serum soluble Gal-9 ELISA were performed on three different cohorts of patients: those with endometriosis, those with benign gynecologic disorders, and healthy controls. Differences in the Gal-9 concentrations between the investigated groups and the stability of Gal-9 in the serum and diagnostic characteristics of Gal-9 ELISA were determined by statistical evaluation and receiver operating characteristic (ROC) curve analysis. Significantly elevated Gal-9 levels were found in both minimal-mild (I-II) and moderate-severe (III-IV) stages of endometriosis in comparison with healthy controls. At a cutoff of 132 pg/mL, ROC analysis revealed an excellent diagnostic value of Gal-9 ELISA in endometriosis (area under the curve = 0.973) with a sensitivity of 94% and specificity of 93.75%, indicating better diagnostic potential than that of other endometriosis biomarkers. Furthermore, various pelvic pain or infertility-associated benign gynecologic conditions were also associated with increased serum Gal-9 levels. Our results suggest that Gal-9 could be a promising noninvasive biomarker of endometriosis and a predictor of various infertility or pelvic pain-related gynecologic disorders.


Coronary artery aneurysm in Kawasaki disease.
A 3-year-old male presents to the emergency department with chief complaints of fever and vomiting. He had a positive rapid streptococcus throat test with cervical lymphadenopathy. The patient was started on antibiotics. On examination, there was diffuse erythematous macular rash on the chest. Laboratory tests revealed elevated white cell count and C-reactive protein. Electrocardiogram was notable for prolonged PR interval indicating 1st degree atrioventricular block. Echocardiogram revealed ectasia of the right coronary artery (RCA). A presumptive diagnosis of Kawasaki disease was made and the patient was started on high-dose aspirin and intravenous immunoglobulins. Cardiac computed tomography angiography (CTA) showed an aneurysm of the proximal RCA measuring up to 7.4 mm. The RCA immediately proximal to the aneurysm measured 3 mm in diameter. The Z score was 13.4. Oblique coronal image from cardiac CTA and volume rendered images demonstrated an aneurysm of the proximal RCA. The patient improved with treatment.


Alterations in lipoprotein metabolism in peroxisome proliferator-activated receptor alpha-deficient mice.
The peroxisome proliferator-activated receptor-alpha (PPARalpha) controls gene expression in response to a diverse class of compounds collectively referred to as peroxisome proliferators. Whereas most known peroxisome proliferators are of exogenous origin and include hypolipidemic drugs and other industrial chemicals, several endogenous PPARalpha activators have been identified such as fatty acids and steroids. The latter finding and the fact that PPARalpha modulates target genes encoding enzymes involved in lipid metabolism suggest a role for PPARalpha in lipid metabolism. This was investigated in the PPARalpha-deficient mouse model. Basal levels of total serum cholesterol, high density lipoprotein cholesterol, hepatic apolipoprotein A-I mRNA, and serum apolipoprotein A-I in PPARalpha-deficient mice are significantly higher compared with wild-type controls. Treatment with the fibrate Wy 14,643 decreased apoA-I serum levels and hepatic mRNA levels in wild-type mice, whereas no effect was detected in the PPARalpha-deficient mice. Administration of the fibrate Wy 14,643 to wild-type mice results in marked depression of hepatic apolipoprotein C-III mRNA and serum triglycerides compared with untreated controls. In contrast, PPARalpha-deficient mice were unaffected by Wy 14,643 treatment. These studies demonstrate that PPARalpha modulates basal levels of serum cholesterol, in particular high density lipoprotein cholesterol, and establish that fibrate-induced modulation in hepatic apolipoprotein A-I, C-III mRNA, and serum triglycerides observed in wild-type mice is mediated by PPARalpha.


Streptococcus sanguis secretes CD14-binding proteins that stimulate cytokine synthesis: a clue to the pathogenesis of infective (bacterial) endocarditis?
Streptococcus sanguis is the major causative organism of infective (bacterial) endocarditis but, surprisingly, almost nothing is known about how it induces endocardial inflammation. In earlier studies we have shown that many bacteria secrete potent cytokine-inducing or -inhibiting proteins. We have therefore isolated the material secreted by S. sanguis grown on blood agar or in broth culture and have tested its ability to induce human peripheral blood monocytes to synthesize pro-inflammatory cytokines. The activation of monocytes by the secreted components of S. sanguis was almost totally blocked by heat and trypsin treatment but not by the lipopolysaccharide-inactivating antibiotic, polymyxin B, suggesting that activity is due to secreted proteins. The activity of the secreted material was significantly reduced by anti-CD14 monoclonal antibodies suggesting that the active protein (or proteins) was binding to the CD14/Toll-like receptor (TLR)4 complex. Fractionation of the secreted proteins by high performance liquid chromatography (HPLC) identified two proteins as being responsible for the majority of the cytokine induction: a manganese-dependent superoxide dismutase and a 190 kDa protein, which could not be sequenced, but which was neither CshA nor the PI/II proteins. These proteins, or the receptors to which they bind, may be therapeutic targets and may allow the development of adjunctive therapies to prevent endocardial damage during the often prolonged treatment of infective endocarditis with antibiotics. In addition, blocking of CD14 may have some therapeutic benefit.


Urethral trauma: radiological aspects and treatment options.
Experience with treatment of 38 patients in an early phase with urethral trauma is presented. The site of injury was the penile urethra in three cases, the bulbous urethra in seven, and the prostatomembranous urethra in 28. Different surgical procedures were used, according to the site and the extent of urethral damage and the presence of associated lesions. All patients with penile and bulbous urethral trauma were cured and only one has diminished sexual potency, while in the posterior urethral group nine were cured, and 19 developed strictures requiring further surgical treatment. Total impotence developed in 17 and partial impotence in one patient. Another case presents urinary urge incontinence.


Sex-dependent and -independent transcriptional changes during haploid phase gametogenesis in the sugar kelp Saccharina latissima.
In haplodiplontic lineages, sexual reproduction occurs in haploid parents without meiosis. Although widespread in multicellular lineages such as brown algae (Phaeophyceae), haplodiplontic gametogenesis has been little studied at the molecular level. We addressed this by generating an annotated reference transcriptome for the gametophytic phase of the sugar kelp, Saccharina latissima. Transcriptional profiles of microscopic male and female gametophytes were analysed at four time points during the transition from vegetative growth to gametogenesis. Gametogenic signals resulting from a switch in culture irradiance from red to white light activated a core set of genes in a sex-independent manner, involving rapid activation of ribosome biogenesis, transcription and translation related pathways, with several acting at the post-transcriptional or post-translational level. Additional genes regulating nutrient acquisition and key carbohydrate-energy pathways were also identified. Candidate sex-biased genes under gametogenic conditions had potentially key roles in controlling female- and male-specific gametogenesis. Among these were several sex-biased or -specific E3 ubiquitin-protein ligases that may have important regulatory roles. Females specifically expressed several genes that coordinate gene expression and/or protein degradation, and the synthesis of inositol-containing compounds. Other female-biased genes supported parallels with oogenesis in divergent multicellular lineages, in particular reactive oxygen signalling via an NADPH-oxidase. Males specifically expressed the hypothesised brown algal sex-determining factor. Male-biased expression mainly involved upregulation of genes that control mitotic cell proliferation and spermatogenesis in other systems, as well as multiple flagella-related genes. Our data and results enhance genome-level understanding of gametogenesis in this ecologically and economically important multicellular lineage.


CTL access to tissue antigen is restricted in vivo.
MHC class I-restricted, hepatitis B surface Ag (HBsAg)-specific, CD8+ polyclonal CTL lines and clones cause a severe necroinflammatory liver disease when they are injected i.v. into transgenic mice that display widespread tissue expression of HBsAg. Surprisingly, the same CTLs fail to cause disease in any other HBsAg-positive tissue in these animals. However, the CTLs are highly cytopathic for HBsAg-positive renal tubules and choroid plexus epithelial cells when they are injected extravascularly, either beneath the kidney capsule or intracerebrally. Analysis of the microvascular anatomy of these tissues reveals that the hepatic sinusoid is characterized by a discontinuous endothelium and the absence of a basement membrane. In contrast, the vasculature of most other tissues displays a continuous endothelium and basement membrane; and the epithelial cells of many of these tissues are also surrounded by a separate basement membrane. These results suggest that viral, tumor, and normal self Ags expressed in the liver are readily accessible to class I-restricted T cells, whereas the vascular endothelium and/or the vascular and cellular basement membranes constitute an extremely effective barrier that precludes CTL access to tissue Ag at many other sites. Because class I-restricted Ag recognition occurs throughout the body in many natural diseases and disease models, the vascular barrier must be breached by other events before CTL access to endogenously synthesized epithelial Ag can occur.


The effect of pranlukast on allergen-induced bone marrow eosinophilopoiesis in subjects with asthma.
We investigated the mechanisms by which leukotriene receptor antagonists decrease airway eosinophil number. In a randomized, double-blind crossover study, we examined the effects of 2 weeks of treatment with pranlukast 300 mg twice a day or placebo on allergen-induced changes in airway eosinophil number and bone marrow eosinophil progenitors in 15 subjects with mild asthma. Pranlukast treatment for 2 weeks decreased mean sputum eosinophil count from 0.15 x 10(6)/g (5.3% of cells) before treatment to 0.02 x 10(6)/g (0.7% of cells) after treatment (p < 0.05), whereas placebo did not. Pranlukast also decreased the eosinophil count (5.6% at 7 hours and 7.5% at 24 hours) (p < 0.05) after allergen inhalation compared with placebo (13.8% at 7 hours and 15.3% at 24 hours). There was a similar trend for sputum cells immunostaining for EG2, eotaxin, interleukin-5, and regulated upon activation, normal T cell expressed and secreted. Pranlukast also significantly attenuated the allergen-induced increase in the number of bone marrow eosinophil/basophil cfu (mean 0.3) at 24 hours compared with placebo (mean 6.2). The proportion of CD34(+) cells expressing the eotaxin receptor CC chemokine receptor 3, 24 hours after allergen inhalation, was also reduced by pranlukast. We conclude that, the cysteinyl leukotriene receptor antagonist, pranlukast, attenuates allergen-induced increase in airway eosinophils by decreasing bone marrow eosinophilopoiesis and airway chemotactic and eosinophilopoietic cytokines.


Computational analysis of high-density peptide microarray data with application from systemic sclerosis to multiple sclerosis.
Auto-antibodies are implicated in the pathophysiology of various autoimmune diseases. High-density peptide microarrays incubated with human serum can detect antibody reactivities against thousands of peptides. This enables the identification of new auto-antigens and the determination of the parts of protein antigens (epitopes) that are recognized by antibody paratopes. We discuss the utility of peptide microarrays to investigate epitope-antibody-recognitions (EAR) from systemic sclerosis to multiple sclerosis. The technology can help to establish reliable diagnostic and prognostic biomarkers employing a combination of antigenic peptides. We describe the specifics of peptide microarray data and present bioinformatic methods for their analysis. Quality control, data pre-processing and the filtering of specific peptides are demonstrated on an example data set. Peptide microarrays representing 24 selected proteins by 3235 overlapping 15mer peptides were used to measure antibodies in serum of 10 patients with limited cutaneous systemic sclerosis (SSC) and 10 healthy blood donors. The data showed a sparse and skewed distribution, and we observed strong individual differences since many peptide sequences were bound by antibodies of only one serum sample. In the sera of the SSc patients, but not of the healthy controls, we found antibodies to three peptides MGPRRRSRKPEAPRR, TPTPGPSRRGPSLGA and GPSRRGPSLGASSHQ that share a similar sequence motif (GP-R/S-RR). These peptides map to two known linear epitopes at the N-terminus of centromere protein A (CENPA), demonstrating the utility of peptide microarrays. Presented experimental and bioinformatic approach can be applied in the same manner for multiple sclerosis research.


Developing a Suicide Prevention Social Media Campaign With Young People (The #Chatsafe Project): Co-Design Approach.
Young people commonly use social media platforms to communicate about suicide. Although research indicates that this communication may be helpful, the potential for harm still exists. To facilitate safe communication about suicide on social media, we developed the #chatsafe guidelines, which we sought to implement via a national social media campaign in Australia. Population-wide suicide prevention campaigns have been shown to improve knowledge, awareness, and attitudes toward suicide. However, suicide prevention campaigns will be ineffective if they do not reach and resonate with their target audience. Co-designing suicide prevention campaigns with young people can increase the engagement and usefulness of these youth interventions. This study aimed to document key elements of the co-design process; to evaluate young people's experiences of the co-design process; and to capture young people's recommendations for the #chatsafe suicide prevention social media campaign. In total, 11 co-design workshops were conducted, with a total of 134 young people aged between 17 and 25 years. The workshops employed commonly used co-design strategies; however, modifications were made to create a safe and comfortable environment, given the population and complexity and sensitivity of the subject matter. Young people's experiences of the workshops were evaluated through a short survey at the end of each workshop. Recommendations for the campaign strategy were captured through a thematic analysis of the postworkshop discussions with facilitators. The majority of young people reported that the workshops were both safe (116/131, 88.5%) and enjoyable (126/131, 96.2%). They reported feeling better equipped to communicate safely about suicide on the web and feeling better able to identify and support others who may be at risk of suicide. Key recommendations for the campaign strategy were that young people wanted to see bite-sized sections of the guidelines come to life via shareable content such as short videos, animations, photographs, and images. They wanted to feel visible in campaign materials and wanted all materials to be fully inclusive and linked to resources and support services. This is the first study internationally to co-design a suicide prevention social media campaign in partnership with young people. The study demonstrates that it is feasible to safely engage young people in co-designing a suicide prevention intervention and that this process produces recommendations, which can usefully inform suicide prevention campaigns aimed at youth. The fact that young people felt better able to safely communicate about suicide on the web as a result of participation in the study augurs well for youth engagement with the national campaign, which was rolled out across Australia. If effective, the campaign has the potential to better prepare many young people to communicate safely about suicide on the web.


Bioremediation efficacy-comparison of nutrient removal from an anaerobic digest waste-based medium by an algal consortium before and after cryopreservation.
An algal consortium was isolated from an integrated steelmaking site at TATA Steel Strip Products Ltd. in Port Talbot, UK, and its bioremediation capacity tested. Excellent "bioremediation" was observed when the mixed culture was "applied" to diluted effluent from an enhanced anaerobic digestion plant at Dŵr Cymru Welsh Water at Port Talbot, UK. After 5 days of cultivation in a 600-L photobioreactor, 99% of the total nitrogen (initial level, 4500 μmol L-1) and total phosphorus (initial level, 690.4 μmol L-1) were removed from the waste stream. The consortium was deposited in the Culture Collection of Algae and Protozoa (CCAP), an international depository authority for microalgal patents, as CCAP 293/1. This material has been successfully cryopreserved using a two-step cryopreservation protocol with dimethyl sulphoxide (5% v/v) used as a cryoprotectant. On recovery of samples after 3 months storage at -196 °C, the specific bioremediation activity of the revived consortium was tested. The capacity of the revived culture to bioremediate effluent was not significantly different (p < 0.05) from a non-cryopreserved control, with 99% of total nitrogen and phosphorus remediated by day 4. Although non-axenic algal cultures have previously been cryopreserved, this is the first report of the successful cryopreservation of mixed algal consortium, with validation of its ability to bioremediate after thawing comparing non-cryopreserved cultures with a revived post-thaw algal consortium. The study also highlights the need to ensure the long-term security and the requirement to validate the functionality of conserved inocula with biotechnological/bioremediation potential.


Consulting communities when patients cannot consent: a multicenter study of community consultation for research in emergency settings.
To assess the range of responses to community consultation efforts conducted within a large network and the impact of different consultation methods on acceptance of exception from informed consent research and understanding of the proposed study. A cognitively pretested survey instrument was administered to 2,612 community consultation participants at 12 U.S. centers participating in a multicenter trial of treatment for acute traumatic brain injury. Survey nested within community consultation for a phase III randomized controlled trial of treatment for acute traumatic brain injury conducted within a multicenter trial network and using exception from informed consent. Adult participants in community consultation events. Community consultation efforts at participating sites. Acceptance of exception from informed consent in general, attitude toward personal exception from informed consent enrollment, and understanding of the study content were assessed. Fifty-four percent of participants agreed exception from informed consent enrollment was acceptable in general in the proposed study; 71% were accepting of personal exception from informed consent enrollment. Participants in interactive versus noninteractive community consultation events were more accepting of exception from informed consent in general (63% vs 49%) and personal exception from informed consent inclusion (77% vs 67%). Interactive community consultation participants had high-level recall of study content significantly more often than noninteractive consultation participants (77% vs 67%). Participants of interactive consultation were more likely to recall possible study benefits (61% vs 45%) but less likely to recall potential risks (56% vs 69%). Interactive community consultation methods were associated with increased acceptance of exception from informed consent and greater overall recall of study information but lower recall of risks. There was also significant variability in exception from informed consent acceptance among different interactive consultation events. These findings have important implications for institutional review board and investigators conducting exception from informed consent research and for community engagement efforts in research more generally.


Rapamycin Interferes With Postdepletion Regulatory T Cell Homeostasis and Enhances DSA Formation Corrected by CTLA4-Ig.
Previously, we demonstrated that alemtuzumab induction with rapamycin as sole maintenance therapy is associated with an increased incidence of humoral rejection in human kidney transplant patients. To investigate the role of rapamycin in posttransplant humoral responses after T cell depletion, fully MHC mismatched hearts were transplanted into hCD52Tg mice, followed by alemtuzumab treatment with or without a short course of rapamycin. While untreated hCD52Tg recipients acutely rejected B6 hearts (n = 12), hCD52Tg recipients treated with alemtuzumab alone or in conjunction with rapamycin showed a lack of acute rejection (MST > 100). However, additional rapamycin showed a reduced beating quality over time and increased incidence of vasculopathy. Furthermore, rapamycin supplementation showed an increased serum donor-specific antibodies (DSA) level compared to alemtuzumab alone at postoperation days 50 and 100. Surprisingly, additional rapamycin treatment significantly reduced CD4(+) CD25(+) FoxP3(+) T reg cell numbers during treatment. On the contrary, ICOS(+) PD-1(+) CD4 follicular helper T cells in the lymph nodes were significantly increased. Interestingly, CTLA4-Ig supplementation in conjunction with rapamycin corrected rapamycin-induced accelerated posttransplant humoral response by directly modulating Tfh cells but not Treg cells. This suggests that rapamycin after T cell depletion could affect Treg cells leading to an increase of Tfh cells and DSA production that can be reversed by CTLA4-Ig.


Environmental context predicts state fluctuations in negative symptoms in youth at clinical high risk for psychosis.
Negative symptoms (avolition, anhedonia, asociality) are a prevalent symptom in those across the psychosis-spectrum and also occur at subclinical levels in the general population. Recent work has begun to examine how environmental contexts (e.g. locations) influence negative symptoms. However, limited work has evaluated how environments may contribute to negative symptoms among youth at clinical high risk for psychosis (CHR). The current study uses Ecological Momentary Assessment to assess how four environmental contexts (locations, activities, social interactions, social interaction method) impact state fluctuations in negative symptoms in CHR and healthy control (CN) participants. CHR youth (n = 116) and CN (n = 61) completed 8 daily surveys for 6 days assessing negative symptoms and contexts. Mixed-effects modeling demonstrated that negative symptoms largely varied across contexts in both groups. CHR participants had higher negative symptoms than CN participants in most contexts, but groups had similar symptom reductions during recreational activities and phone call interactions. Among CHR participants, negative symptoms were elevated in several contexts, including studying/working, commuting, eating, running errands, and being at home. Results demonstrate that negative symptoms dynamically change across some contexts in CHR participants. Negative symptoms were more intact in some contexts, while other contexts, notably some used to promote functional recovery, may exacerbate negative symptoms in CHR. Findings suggest that environmental factors should be considered when understanding state fluctuations in negative symptoms among those at CHR participants.


Molecular medicine: a primer for clinicians--Part VIII: Forensic DNA testing.
Forensic DNA analysis or "DNA fingerprinting" represents a specific application of DNA testing methods. Previously, we have discussed DNA testing to determine carrier status or to provide confirmatory or presymptomatic diagnosis. Forensic DNA testing seeks to determine the identity of an individual to the exclusion of all others. The most common application of forensic DNA testing is in criminalistics and establishing parentage. In this paper we discuss the application of molecular biology methods to the analysis of DNA for forensic purposes. We also consider some of the controversial issues that surround such uses of DNA testing.


Expression of Bacteroides fragilis hemolysins in vivo and role of HlyBA in an intra-abdominal infection model.
Bacteroides fragilis is the most frequent opportunistic pathogen isolated from anaerobic infections. However, there is a paucity of information regarding the genetic and molecular aspects of gene expression of its virulence factors during extra-intestinal infections. A potential virulence factor that has received little attention is the ability of B. fragilis to produce hemolysins. In this study, an implanted perforated table tennis "ping-pong" ball was used as an intra-abdominal artificial abscess model in the rat. This procedure provided sufficient infected exudate for gene expression studies in vivo. Real-time reverse transcription polymerase chain reaction (RT-PCR) was used to quantify the relative expression of hlyA, hlyB, hlyC, hlyD, hlyE, hlyF, hlyG, and hlyIII mRNAs. The hlyA mRNA was induced approximately sixfold after 4 days postinfection compared with the mRNA levels in the inoculum culture prior to infection. The hlyB mRNA increased approximately sixfold after 4 days and 12-fold after 8 days postinfection. Expression of hlyC mRNA increased sixfold after 1 day, 45-fold after 4 days, and 16-fold after 8 days postinfection, respectively. The hlyD and hlyE mRNAs were induced approximately 40-fold and 30-fold, respectively, after 4-days postinfection. The hlyF expression increased approximately threefold after 4-days postinfection. hlyG was induced approximately fivefold after 4 and 8 days postinfection. The hlyIII mRNA levels had a steady increase of approximately four-, eight-, and 12-fold following 1, 4, and 8 days postinfection, respectively. These findings suggest that B. fragilis hemolysins are induced and differentially regulated in vivo. Both parent and hlyBA mutant strains reached levels of approximately 3-8 × 10(9) cfu/mL after 1 day postinfection. However, the hlyBA mutant strain lost 2 logs in viable cell counts compared with the parent strain after 8 days postinfection. This is the first study showing HlyBA is a virulence factor which plays a role in B. fragilis survival in an intra-abdominal abscess model.


Expression of epiregulin and amphiregulin and K-ras mutation status predict disease control in metastatic colorectal cancer patients treated with cetuximab.
The antiepidermal growth factor receptor (EGFR) antibody cetuximab shows activity in multiple epithelial tumor types; however, responses are seen in only a subset of patients. This study was conducted to identify markers that are associated with disease control in patients treated with cetuximab. One hundred ten patients with metastatic colorectal cancer were enrolled onto a cetuximab monotherapy trial. Transcriptional profiling was conducted on RNA from mandatory pretreatment metastatic biopsies to identify genes whose expression correlates with best clinical responses. EGFR and K-ras mutation analyses and EGFR gene copy number analyses were performed on DNA from pretreatment biopsies. Gene expression profiles showed that patients with tumors that express high levels of the EGFR ligands epiregulin and amphiregulin are more likely to have disease control with cetuximab (EREG, P = .000015; AREG, P = .000025). Additionally, patients whose tumors do not have K-ras mutations have a significantly higher disease control rate than patients with K-ras mutations (P = .0003). Furthermore, patients with tumors that have high expression of EREG or AREG also have significantly longer progression-free survival (PFS) than patients with low expression (EREG: P = .0002, hazard ratio [HR] = 0.47, and median PFS, 103.5 v 57 days, respectively; AREG: P < .0001, HR = 0.44, and median PFS, 115.5 v 57 days, respectively). Patients with tumors that have high gene expression levels of epiregulin and amphiregulin and patients with wild-type K-ras are more likely to have disease control on cetuximab treatment. The identified markers could be developed further to select patients for cetuximab therapy.


Spread of Plasmid-Encoded NDM-1 and GES-5 Carbapenemases among Extensively Drug-Resistant and Pandrug-Resistant Clinical Enterobacteriaceae in Durban, South Africa.
Whole-genome sequence analyses revealed the presence of blaNDM-1 (n = 31), blaGES-5 (n = 8), blaOXA-232 (n = 1), or blaNDM-5 (n = 1) in extensively drug-resistant and pandrug-resistant Enterobacteriaceae organisms isolated from in-patients in 10 private hospitals (2012 to 2013) in Durban, South Africa. Two novel NDM-1-encoding plasmids from Klebsiella pneumoniae were circularized by PacBio sequencing. In p19-10_01 [IncFIB(K); 223.434 bp], blaNDM-1 was part of a Tn1548-like structure (16.276 bp) delineated by IS26 The multireplicon plasmid p18-43_01 [IncR_1/IncFIB(pB171)/IncFII(Yp); 212.326 bp] shared an 80-kb region with p19-10_01, not including the blaNDM-1-containing region. The two plasmids were used as references for tracing NDM-1-encoding plasmids in the other genome assemblies. The p19-10_01 sequence was detected in K. pneumoniae (n = 7) only, whereas p18-43_01 was tracked to K. pneumoniae (n = 4), Klebsiella michiganensis (n = 1), Serratia marcescens (n = 11), Enterobacter spp. (n = 7), and Citrobacter freundii (n = 1), revealing horizontal spread of this blaNDM-1-bearing plasmid structure. Global phylogeny showed clustering of the K. pneumoniae (18/20) isolates together with closely related carbapenemase-negative ST101 isolates from other geographical origins. The South African isolates were divided into three phylogenetic subbranches, where each group had distinct resistance and replicon profiles, carrying either p19-10_01, p18-10_01, or pCHE-A1 (8,201 bp). The latter plasmid carried blaGES-5 and aacA4 within an integron mobilization unit. Our findings imply independent plasmid acquisition followed by local dissemination. Additionally, we detected blaOXA-232 carried by pPKPN4 in K. pneumoniae (ST14) and blaNDM-5 contained by a pNDM-MGR194-like genetic structure in Escherichia coli (ST167), adding even more complexity to the multilayer molecular mechanisms behind nosocomial spread of carbapenem-resistant Enterobacteriaceae in Durban, South Africa.


Effectiveness of Antipseudomonal Antibiotics and Mechanisms of Multidrug Resistance in Pseudomonas aeruginosa.
Pseudomonas aeruginosa is a leading human pathogen that causes serious infections at various tissues and organs leading to life threatening health problems and possible deadly outcomes. Resistance patterns vary widely whether it is from hospitals or community acquired infections. Reporting resistance profiles to a certain antibiotics provide valuable information in a given setting, but may be extrapolated outside the sampling location. In the present study, P. aeruginosa isolates were screened to determine their susceptibilities against anti-pseudomonal antimicrobial agents and possible existing mechanisms of resistance were determined. Eighty-six isolates of P. aeruginosa were recovered. Isolates representing different resistance profiles were screened for the existence of three different resistance mechanisms including drug inactivation due to metallo-β-lactamases, drug impermeability by outer membrane proteins and drug efflux. All tested isolates showed uniform susceptibility (100%, n = 86/86) to piperacillin, meropenem, amikacin, and polymyxin B. A single isolate was found to be imipenem resistant (99%, n = 85/86). The possible mechanisms of resistance of P. aeruginosa to imipenem involve active drug efflux pumps, outer membrane impermeability as well as drug inactivating enzymes. These findings demonstrate the fundamental importance of the in vitro susceptibility testing of antibiotics prior to antipseudomonal therapy and highlight the need for a continuous antimicrobial resistance surveillance programs to monitor the changing resistance patterns so that clinicians and health care officials are updated as to the most effective therapeutic agents to combat the serious outcomes of P. aeruginosa infections.


In vitro formation of osteoclasts from long-term cultures of bone marrow mononuclear phagocytes.
The origin of osteoclasts was studied in an in vitro model using organ cultures of periosteum-free embryonic mouse long-bone primordia, which were co-cultured with various cell populations. The bone rudiments were freed of their periosteum-perichondrium by collagenase treatment in a stage before cartilage erosion and osteoclast formation, and co-cultured for 7 d with either embryonic liver or mononuclear phagocytes from various sources. Light and electron microscopic examination of the cultures showed that mineralized matrix-resorbing osteoclasts developed only in bones co-cultured with embryonic liver or with cultured bone marrow mononuclear phagocytes but not when co-cultured with blood monocytes or resident or exudate peritoneal macrophages. Osteoclasts developed from the weakly adherent, but not from the strongly adherent cells of bone marrow cultures, whereas 1,000 rad irradiation destroyed the capacity of such cultures to form osteoclasts. In bone cultures to which no other cells were added, osteoclasts were virtually absent. Bone-resorbing activity of in vitro formed osteoclasts was demonstrated by 45Ca release studies. These studies demonstrate that osteoclasts develop from cells present in cultures of proliferating mononuclear phagocytes and that, at least in our system, monocytes and macrophages are unable to form osteoclasts. The most likely candidates for osteoclast precursor cells seem to be monoblasts and promonocytes.