Characterisation and classification of phylloplane yeasts from Portugal related to the genus Taphrina and description of five novel Lalaria species.
Taphrina Fries is a genus of dimorphic ascomycetes comprising more than 90 species distinguished by the specific infections they produce on different vascular plants. Their filamentous states are restricted to parasitised plant tissue whereas the yeast states are saprobic and can be grown on artificial media. The latter coincide with the anamorphic phases and have been given separate nomenclatural status by the erection of the genus Lalaria R.T. Moore. In its original circumscription Lalaria included only 23 yeast states of known species of Taphrina and its creation was then redundant. Here we describe five novel species in the genus Lalaria to accommodate a total of 44 yeast isolates obtained mainly from leaf surfaces (phylloplane) of different plants in Portugal: Lalaria arrabidae sp. nov. (one strain), L. carpini sp. nov. (one strain), L. inositophila sp. nov. (37 strains), L. kurtzmanii sp. nov. (one strain) and L. veronaerambellii sp. nov. (four strains). L. inositophila was notable for its widespread occurrence since it was recovered during two consecutive years from the leaves of miscellaneous plant species. In the absence of sexual states and of unequivocal associations to particular host plants, the taxonomic relationship of the novel species to the yeast states of Taphrina available from culture collections was verified by the comparative analysis of physiological and molecular characteristics. The latter included PCR fingerprinting using single primers for microsatellite regions, sequencing of the 5' end of the 26S rRNA (LSU) gene (D1/D2 domains) and of the ITS1 and ITS2 rDNA spacer regions, and DNA-DNA hybridisation experiments. An emended description of the genus Lalaria is provided.


Healthcare providers on the frontlines: a qualitative investigation of the social and emotional impact of delivering health services during Sierra Leone's Ebola epidemic.
Although research on the epidemiology and ecology of Ebola has expanded since the 2014-15 outbreak in West Africa, less attention has been paid to the mental health implications and the psychosocial context of the disease for providers working in primary health facilities (rather than Ebola-specific treatment units). This study draws on 54 qualitative interviews with 35 providers working in eight peripheral health units of Sierra Leone's Bo and Kenema Districts. Data collection started near the height of the outbreak in December 2014 and lasted 1 month. Providers recounted changes in their professional, personal and social lives as they became de facto first responders in the outbreak. A theme articulated across interviews was Ebola's destruction of social connectedness and sense of trust within and across health facilities, communities and families. Providers described feeling lonely, ostracized, unloved, afraid, saddened and no longer respected. They also discussed restrictions on behaviors that enhance coping including attending burials and engaging in physical touch (hugging, handshaking, sitting near, or eating with colleagues, patients and family members). Providers described infection prevention measures as necessary but divisive because screening booths and protective equipment inhibited bonding or 'suffering with' patients. To mitigate psychiatric morbidities and maladaptive coping mechanisms-and to prevent the spread of Ebola-researchers and program planners must consider the psychosocial context of this disease and mechanisms to enhance psychological first aid to all health providers, including those in peripheral health settings.


Genome-wide association study of serious blistering skin rash caused by drugs.
Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) are rare but severe, potentially life threatening adverse drug reactions characterized by skin blistering. Previous studies have identified drug-specific and population-specific genetic risk factors with large effects. In this study, we report the first genome-wide association study (GWAS) of SJS/TEN induced by a variety of drugs. Our aim was to identify common genetic risk factors with large effects on SJS/TEN risk. We conducted a genome-wide analysis of 96 retrospective cases and 198 controls with a panel of over one million single-nucleotide polymorphisms (SNPs). We further improved power with about 4000 additional controls from publicly available datasets. No genome-wide significant associations with SNPs or copy number variants were observed, although several genomic regions were suggested that may have a role in predisposing to drug-induced SJS/TEN. Our GWAS did not find common, highly penetrant genetic risk factors responsible for SJS/TEN events in the cases selected.


Insomnia and posttraumatic stress symptoms: Evidence of shared etiology.
Posttraumatic stress disorder (PTSD) and insomnia are comorbid clinical conditions that are thought to result from genetic and environmental effects. Though studies have established the heritability of these disorders independently, no study to date has examined the genetic contributions to the relation between insomnia and PTSD symptoms (PTSS). The present study assessed this gap in the literature using a behavioral genetics approach to symptom dimensions. The sample consisted of 242 twin pairs who endorsed lifetime trauma exposure. Insomnia symptoms were assessed with the Women's Health Initiative Survey, and intrusion and avoidance PTSS were assessed with the Impact of Events Scale. Structural equation modeling was then employed to test the relative contributions of genetic, shared environmental, and nonshared environmental components to the relations between insomnia symptoms and intrusions and avoidance. Results indicated a significant association between insomnia symptoms and intrusions (r = 0.33, p < 0.01) and insomnia symptoms and avoidance (r = 0.20, p < 0.01), and 36-44% of phenotypic variance was accounted for by genetic contributions. These findings highlight a significant role for genetic factors in the mechanisms underlying the comorbidity between insomnia and PTSS. The implications for current etiological models of PTSD and insomnia are discussed.


Use of item response theory and latent class analysis to link poly-substance use disorders with addiction severity, HIV risk, and quality of life among opioid-dependent patients in the Clinical Trials Network.
This study applied item response theory (IRT) and latent class analysis (LCA) procedures to examine the dimensionality and heterogeneity of comorbid substance use disorders (SUDs) and explored their utility for standard clinical assessments, including the Addiction Severity Index (ASI), HIV Risk Behavior Scale (HRBS), and SF-36 quality-of-life measures. The sample included 343 opioid-dependent patients enrolled in two national multisite studies of the U.S. National Drug Abuse Treatment Clinical Trials Network (CTN001-002). Patients were recruited from inpatient and outpatient addiction treatment settings at 12 programs. Data were analyzed by factor analysis, IRT, LCA, and latent regression procedures. A two-class LCA model fit dichotomous SUD data empirically better than one-parameter and two-parameter IRT models. LCA distinguished 10% of severe comorbid opioid-dependent individuals who had high rates of all SUDs examined-especially amphetamine and sedative abuse/dependence-from the remaining 90% who had SUDs other than amphetamine and sedative abuse/dependence (entropy=0.99). Item-level results from both one-parameter and two-parameter IRT models also found that amphetamine and sedative abuse/dependence tapped the more severe end of the latent poly-SUD trait. Regardless of whether SUDs were defined as a continuous trait or categorically, individuals characterized by a high level of poly-SUD demonstrated more psychiatric problems and HIV risk behaviors. A combined application of categorical and dimensional latent approaches may improve the understanding of comorbid SUDs and their associations with other clinical indicators. Abuse of sedatives and methamphetamine may serve as a useful marker for identifying subsets of opioid-dependent individuals with needs for more intensive interventions.


Defining a land boundary for sustainable livestock consumption.
The need for more sustainable production and consumption of animal source food (ASF) is central to the achievement of the sustainable development goals: within this context, wise use of land is a core challenge and concern. A key question in feeding the future world is: how much ASF should we eat? We demonstrate that livestock raised under the circular economy concept could provide a significant, nonnegligible part (9-23 g/per capita) of our daily protein needs (~50-60 g/per capita). This livestock then would not consume human-edible biomass, such as grains, but mainly convert leftovers from arable land and grass resources into valuable food, implying that production of livestock feed is largely decoupled from arable land. The availability of these biomass streams for livestock then determines the boundaries for livestock production and consumption. Under this concept, the competition for land for feed or food would be minimized and compared to no ASF, including some ASF in the human diet could free up about one quarter of global arable land. Our results also demonstrate that restricted growth in consumption of ASF in Africa and Asia would be feasible under these boundary conditions, while reductions in the rest of the world would be necessary to meet land use sustainability criteria. Managing this expansion and contraction of future consumption of ASF is essential for achieving sustainable nutrition security.


Activity-dependent suppression of miniature neurotransmission through the regulation of DNA methylation.
DNA methylation is an epigenetic mechanism that plays a critical role in the repression of gene expression. Here, we show that DNA methyltransferase (DNMT) inhibition in hippocampal neurons results in activity-dependent demethylation of genomic DNA and a parallel decrease in the frequency of miniature EPSCs (mEPSCs), which in turn impacts neuronal excitability and network activity. Treatment with DNMT inhibitors reveals an activity-driven demethylation of brain-derived neurotrophic factor promoter I, which is mediated by synaptic activation of NMDA receptors, because it is susceptible to AP-5, a blocker of NMDA receptors. The specific effect of DNMT inhibition on spontaneous excitatory neurotransmission requires gene transcription and is occluded in the absence of the transcriptional repressor methyl-CpG-binding protein 2 (MeCP2). Interestingly, enhancing excitatory activity, in the absence of DNMT inhibitors, also produces similar decreases in DNA methylation and mEPSC frequency, suggesting a role for DNA methylation in the control of homeostatic synaptic plasticity. Furthermore, adding excess substrate for DNA methylation (S-adenosyl-L-methionine) rescues the suppression of mEPSCs by DNMT inhibitors in wild-type neurons, as well as the defect seen in MeCP2-deficient neurons. These results uncover a means by which NMDA receptor-mediated synaptic activity drives DNA demethylation within mature neurons and suppresses basal synaptic function.


Military Personnel Ratings of a Deployment and Their Positive and Negative Deployment Experiences.
There is limited investigation of how military personnel evaluate their deployment experiences. An understanding of their perceptions would help unit psychologists to advise commanders on ways to improve the deployment experience (and therefore mental well-being) of personnel. This study examined the interplay between deployment overall ratings, personnel characteristics and positive and negative deployment experiences in aid of such understanding. The participants were 1,226 Australian Defence Force personnel who deployed to East Timor and (through a survey) provided an overall rating of their deployment and comments on major positive and negative deployment experiences. Descriptive statistics detail ratings by personnel characteristics, and a hybrid content/thematic analysis details the positive and negative experiences. The study was approved by an Institutional Review Board. Over 80% of the participants rated their overall East Timor deployment experience as positive, with 13% rating it as neutral and 7% as negative. Intrinsic rewards (eg, ability to use skills) were the most commonly expressed major positive experiences of the deployment, with deployment administration and military leadership the most common negatives. Most intrinsic rewards were reported more often in participants with a positive deployment rating, while poor leadership was most frequent in those with a negative rating. Military leadership is corroborated as a negative experience of military deployment, while a new finding indicates that intrinsic rewards are a common feature in positive evaluations of deployment. Leadership is a factor that Defence Forces can address to improve the deployment experience. The study strength is the range and size of the sample, with a limitation the potential for recall bias (the data were collected, on average, 5 years postdeployment). Future research should replicate this type of analysis to build a picture of the experiences and evaluations of personnel from a range of different deployments.


Integrating fluorescent molecularly imprinted polymer (MIP) sensor particles with a modular microfluidic platform for nanomolar small-molecule detection directly in aqueous samples.
Fluorescent sensory MIP (molecularly imprinted polymer) particles were combined with a droplet-based 3D microfluidic system for the selective determination of a prototype small-molecule analyte of environmental concern, 2,4-dichlorophenoxyacetic acid or 2,4-D, at nanomolar concentration directly in water samples. A tailor-made fluorescent indicator cross-linker was thus designed that translates the binding event directly into an enhanced fluorescence signal. The phenoxazinone-type cross-linker was co-polymerized into a thin MIP layer grafted from the surface of silica microparticles following a RAFT (reversible addition-fragmentation chain transfer) polymerization protocol. While the indicator cross-linker outperformed its corresponding monomer twin, establishment of a phase-transfer protocol was essential to guarantee that the hydrogen bond-mediated signalling mechanism between the urea binding site on the indicator cross-linker and the carboxylate group of the analyte was still operative upon real sample analysis. The latter was achieved by integration of the fluorescent core-shell MIP sensor particles into a modular microfluidic platform that allows for an in-line phase-transfer assay, extracting the analyte from aqueous sample droplets into the organic phase that contains the sensor particles. Real-time fluorescence determination of 2,4-D down to 20nM was realized with the system and applied for the analysis of various surface water samples collected from different parts of the world.


Osteogenesis at the dental implant interface: high-voltage electron microscopic and conventional transmission electron microscopic observations.
The osteogenesis of mandibular bone to endosteal dental implants was examined using an in vivo dog model. One half of the implants examined were unloaded implants, with the remaining one half prosthodontically loaded for 6 months. Undecalcified mandibular implant samples were examined with both high-voltage electron microscopy (HVEM) stereology and routine transmission electron microscopy. The osseous interface to integrated implants was shown to vary in its morphology. Mineralized bone was observed directly apposing the implant, often separated from the implant by an electron-dense deposit of approximately 50 nm. Within this densely mineralized matrix, osteocytes were routinely observed. Adjacent areas were shown to contain slightly wider zones of either a less dense mineralized matrix or, alternatively, unmineralized tissue. Other zones consisted of wider unmineralized matrices containing collagen fibers and osteoblasts. These latter zones were consistent with the appearance of an appositional type of bone growth. Because bone is a dynamic, actively remodeling tissue, a varied morphology of the support tissues to dental implant is not unexpected. Areas of mature bone interfacing with successfully integrated implants were demonstrated, as well as areas adjacent to the mature bone that were undergoing remodeling or mineralization. This study has also shown that HVEM stereology is a valuable research tool to investigate the oral tissue interface with dental implants.


The influence of a family program on adolescent tobacco and alcohol use.
This study examined a family-directed program's effectiveness in preventing adolescent tobacco and alcohol use in a general population. Adolescents aged 12 to 14 years and their families were identified by random-digit dialing throughout the contiguous United States. After providing baseline data by telephone interviews, they were randomly allocated to receive or not receive a family-directed program featuring mailed booklets and telephone contacts by health educators. Follow-up telephone interviews were conducted 3 and 12 months after program completion. The findings suggested that smoking onset was reduced by 16.4% at 1 year, with a 25.0% reduction for non-Hispanic Whites but no statistically significant program effect for other races/ethnicities. There were no statistically significant program effects for smokeless tobacco or alcohol use onset. The family-directed program was associated with reduced smoking onset for non-Hispanic Whites, suggesting that it is worthy of further application, development, and evaluation.


Role of histone acetylation in the stimulatory effect of valproic acid on vascular endothelial tissue-type plasminogen activator expression.
Stimulated release of tissue-type plasminogen activator (t-PA) is pivotal for an intravascular fibrinolytic response and protects the circulation from occluding thrombosis. Hence, an impaired t-PA production is associated with increased risk for atherothrombotic events. A pharmacological means to stimulate the production of this enzyme may thus be desirable. We investigated if the anti-epileptic drug valproic acid (VPA) is capable of enhancing t-PA expression in vitro in vascular endothelial cells, and further examined if its histone deacetylase (HDAC)-inhibitory activity is of importance for regulating t-PA expression. Human endothelial cells were exposed to valproic acid and t-PA mRNA and protein levels were quantified. Potential changes in histone acetylation status globally and at the t-PA promoter were examined by western blot and chromatin immunoprecipitation. Valproic acid dose-dependently stimulated t-PA mRNA and protein expression in endothelial cells reaching a 2-4-fold increase at clinically relevant concentrations and 10-fold increase at maximal concentrations. Transcription profiling analysis revealed that t-PA is selectively targeted by this agent. Augmented histone acetylation was detected at the t-PA transcription start site, and an attenuated VPA-response was observed with siRNA knock of HDAC3, HDAC5 and HDAC7. Valproic acid induces t-PA expression in cultured endothelial cells, and this is associated with increased histone acetylation at the t-PA promoter. Given the apparent potency of valproic acid in stimulating t-PA expression in vitro this substance may be a candidate for pharmacological modulation of endogenous fibrinolysis in man.


Metal concentrations in the eggs of endangered Florida scrub-jays from central Florida.
We analyzed metal levels in the eggs of the endangered Florida scrub-jay (Aphelocoma coerulescens), an extremely sedentary and modestly long-lived passerine bird. Eggs were obtained as part of the long-term study of this species at the Archbold Biological Station, located in south central Florida. Following a brief and usually short dispersal, Florida scrub-jays remain in a permanent territory in their scrub oak habitat, where they feed mainly on insects and acorns. As they are low on the food chain and sedentary, we expected levels of contaminants to be comparatively low and to reflect the local environment. No significant yearly differences existed for any metal between eggs collected in 1997 (n = 11) and 1998 (n = 5). Arithmetic mean metal concentrations (whole egg, dry weight) were low for arsenic (34 ppb), cadmium (25 ppb), mercury (74 ppb), and lead (66 ppb), intermediate for chromium (226 ppb), and higher for selenium (1,470 ppb) and manganese (2,230 ppb). Except for the low mercury value, all these means were within the range of mean values reported for a wide range of nonpasserine species, including those at high trophic levels, but are lower than concentrations associated with abnormalities in birds. The three essential elements (chromium, selenium, and manganese) were approximately normally distributed, whereas arsenic, cadmium, lead, and mercury were highly skewed (arithmetic mean much higher than geometric mean).


Sodium hydrosulfide inhibits the differentiation of osteoclast progenitor cells via NRF2-dependent mechanism.
Hydrogen sulfide (H2S), which recently emerged as a potent regulator of tissues and organs, is broadly produced in mammalian cells but whether it can regulate bone cell function is still elusive. The main objective of this study was to establish the role of H2S in the regulation of human osteoclast differentiation and function. Sodium hydrosulfide (NaHS), a common H2S-donor, was administered in vitro to CD11b+ human monocytes, the pool of circulating osteoclasts precursors which are critically involved in osteoclast development and function in bone. NaHS dose-dependently decreased human osteoclast differentiation at concentrations which did not induce toxicity. The inhibition of human osteoclast differentiation was associated with a down-regulation in RANKL-dependent intracellular ROS levels in human pre-osteoclasts cells. Furthermore, NaHS up-regulated NRF2 protein expression, its nuclear translocation, and the transcription of the two key downstream antioxidant genes Peroxiredoxin-1 and NAD(P)H dehydrogenase quinone 1, suggesting that NRF2 activation may inhibit human osteoclast differentiation by activating a sustained antioxidant response in osteoclast progenitors; furthermore, NRF2 activators Sulforaphane and Tert-butylhydroquinone inhibited in vitro human osteoclast differentiation. Moreover, silencing NRF2 in human pre-osteoclasts totally abolished NaHS-mediated inhibition of osteoclastogenesis, suggesting that NRF2 is essential to the inhibitory function of NaHS in osteoclast development. Finally, we found that NaHS also downregulated the RANKL/OPG mRNA ratio in human mesenchymal stem cells, the key osteoclast-supporting cells. Our results suggest that NaHS shows a potential therapeutical role in erosive diseases of bone by regulating both direct and indirect mechanisms controlling the differentiation of circulating osteoclasts precursors.


Expression of transcription factor Yin Yang 1 in prostate cancer.
The transcription repressor Yin Yang 1 (YY1) is expressed in several human cancer cell lines and its expression correlates with resistance to immune-mediated apoptosis. This study used tissue microarrays to investigate the expression and localization of YY1 in 1364 representative tissue samples from 246 hormone naive prostate cancer patients who underwent radical prostatectomy. Staining intensity and frequency measures for both YY1 nuclear and cytoplasmic expression were higher in neoplastic tissues and in PIN samples compared to matched benign cells (p < 0.0001 for all comparisons). Expression of YY1 is predominantly elevated in early malignancy (PIN), as well as in tumors of intermediate to high morphologic grade (Gleason's grade 3-5). Using multivariate Cox proportional hazards analysis, we observed that low nuclear YY1 staining is an independent predictor of a shorter time to recurrence (p = 0.012). Based on these results, we hypothesize that YY1 may play a role in prostate cancer development; however, decreased YY1 may give metastatic cells a survival advantage. These results may also implicate YY1 as a useful diagnostic and prognostic marker.


An oligodendrocyte-specific zinc-finger transcription regulator cooperates with Olig2 to promote oligodendrocyte differentiation.
Molecular mechanisms that control oligodendrocyte myelination during mammalian central nervous system (CNS) development are poorly understood. In this study, we identified Zfp488, an oligodendrocyte-specific zinc-finger transcription regulator, by screening for genes downregulated in the optic nerves of Olig1-null mice. The predicted primary structure of Zfp488 is evolutionarily conserved in vertebrates and invertebrates. In the developing CNS, Zfp488 is specifically expressed in oligodendrocytes but not their precursors. Its expression increases in parallel with that of major myelin genes Mbp and Plp1. Zfp488 is a nuclear protein that possesses transcriptional repression activity. In the developing chick neural tube, Zfp488 can promote oligodendrocyte precursor formation upon Notch activation. In addition, Zfp488 can interact and cooperate with the bHLH transcription factor Olig2 to promote precocious and ectopic oligodendrocyte differentiation. Furthermore, knockdown of Zfp488 via RNAi in an oligodendroglial cell line leads to the downregulation of myelin gene expression. Taken together, these data suggest that Zfp488 functions as an oligodendrocyte-specific transcription co-regulator important for oligodendrocyte maturation and that zinc-finger/bHLH cooperation can serve as a mechanism for oligodendroglial differentiation.


Effect of levodopa priming on dopamine neuron transplant efficacy and induction of abnormal involuntary movements in parkinsonian rats.
Clinical trials of neural grafting for Parkinson's disease (PD) have produced variable, but overall disappointing, results. One particular disappointment has been the development of aberrant motor complications following dopamine (DA) neuron grafting. Despite a lack of consistent benefit, the utility of dopamine neuron replacement remains supported by clinical and basic data. In a continued effort to elucidate factors that might improve this therapy, we used a parkinsonian rat model to examine whether pregraft chronic levodopa affected graft efficacy and/or graft-induced dyskinesia (GID) induction. Indeed, all grafted PD patients to date have had a pregraft history of long-term levodopa. It is well established that long-term levodopa results in a plethora of long-lasting neurochemical alterations and genomic changes indicative of altered structural and synaptic plasticity. Thus, therapeutic dopamine terminal replacement in a striatal environment complicated by such changes could be expected to lead to abnormal or inappropriate connections between graft and host brain and to contribute to suboptimal efficacy and/or postgraft GID behaviors. To investigate the effect of pregraft levodopa, one group of parkinsonian rats received levodopa for 4 weeks prior to grafting. A second levodopa-naïve group was grafted, and the grafts were allowed to mature for 9 weeks prior to introducing chronic levodopa. We report here that, in parkinsonian rats, preexposure to chronic levodopa significantly reduces behavioral and neurochemical efficacy of embryonic dopamine grafts. Furthermore, dopamine terminal replacement prior to introduction of chronic levodopa is highly effective at preventing development of levodopa-induced dyskinesias, and GID-like behaviors occur regardless of pregraft levodopa status.


Work group IV: Future directions for measures of the food and physical activity environments.
Much progress has been made in the past 5 to 10 years in measuring and understanding the impact of the food and physical activity environments on behavioral outcomes. Nevertheless, this research is in its infancy. A work group was convened to identify current evidence gaps and barriers in food and physical activity environments and policy research measures, and develop recommendations to guide future directions for measurement and methodologic research efforts. A nominal group process was used to determine six priority areas for food and physical activity environments and policy measures to move the field forward by 2015, including: (1) identify relevant factors in the food and physical activity environments to measure, including those most amenable to change; (2) improve understanding of mechanisms for relationships between the environment and physical activity, diet, and obesity; (3) develop simplified measures that are sensitive to change, valid for different population groups and settings, and responsive to changing trends; (4) evaluate natural experiments to improve understanding of food and physical activity environments and their impact on behaviors and weight; (5) establish surveillance systems to predict and track change over time; and (6) develop standards for adopting effective health-promoting changes to the food and physical activity environments. The recommendations emanating from the work group highlight actions required to advance policy-relevant research related to food and physical activity environments.


Comparisons of nerve fiber layer thickness measurements between Stratus, Cirrus, and RTVue OCTs in healthy and glaucomatous eyes.
To compare retinal nerve fiber layer (RNFL) measurements obtained by two spectral-domain optical coherence tomographies (SD OCTs; Cirrus-HD OCT and RTVue) and a time domain OCT (TD OCT; Stratus OCT). The diagnostic ability of the three instruments for glaucoma has also been compared. One hundred eight open-angle glaucoma patients and 46 normal controls were enrolled. Three OCT scans were repeated to obtain peripapillary RNFL thickness measurements from all participants on the same day. The relationships between RNFL measurements from three OCTs were evaluated with regression analyses. The thickest RNFL measurements were generally obtained with the RTVue, followed by the Stratus, and finally by the Cirrus OCT (mixed model, p < 0.05). However, the tendency was reversed or no longer present in severe glaucomatous eyes and nasal quadrant maps. Regression analysis between the TD OCT and the two SD OCTs revealed a quadratic relation (linear vs. non-linear, Akaike information criterion, all p < 0.05), whereby RNFL measurements of the TD OCT became exponentially thinner than those by the SD OCTs in thin RNFL thicknesses. Comparing the two SD OCTs, RTVue measured thicker values consistently regardless of the RNFL thickness (pairwise comparisons, p < 0.05). There were no significant differences in the diagnostic ability of all three instruments in average and each quadrants of RNFL thickness (p > 0.05). Direct comparisons of RNFL thickness measurements among OCT instruments may be misleading as there are considerable differences among devices. The measurement differences between SD OCT and TD OCT differed according to the RNFL thickness and also possibly to severity of the glaucoma. The diagnostic ability of each instrument for glaucoma detection was not significantly different.


Characterization of Mycosphaerellaceae species associated with citrus greasy spot in Panama and Spain.
Greasy spot of citrus, caused by Zasmidium citri-griseum (= Mycosphaerella citri), is widely distributed in the Caribbean Basin, inducing leaf spots, premature defoliation, and yield loss. Greasy spot-like symptoms were frequently observed in humid citrus-growing regions in Panama as well as in semi-arid areas in Spain, but disease aetiology was unknown. Citrus-growing areas in Panama and Spain were surveyed and isolates of Mycosphaerellaceae were obtained from citrus greasy spot lesions. A selection of isolates from Panama (n = 22) and Spain (n = 16) was assembled based on their geographical origin, citrus species, and affected tissue. The isolates were characterized based on multi-locus DNA (ITS and EF-1α) sequence analyses, morphology, growth at different temperatures, and independent pathogenicity tests on the citrus species most affected in each country. Reference isolates and sequences were also included in the analysis. Isolates from Panama were identified as Z. citri-griseum complex, and others from Spain attributed to Amycosphaerella africana. Isolates of the Z. citri-griseum complex had a significantly higher optimal growth temperature (26.8°C) than those of A. africana (19.3°C), which corresponded well with their actual biogeographical range. The isolates of the Z. citri-griseum complex from Panama induced typical greasy spot symptoms in 'Valencia' sweet orange plants and the inoculated fungi were reisolated. No symptoms were observed in plants of the 'Ortanique' tangor inoculated with A. africana. These results demonstrate the presence of citrus greasy spot, caused by Z. citri-griseum complex, in Panama whereas A. africana was associated with greasy spot-like symptoms in Spain.


Role of the community matron in advance care planning and 'do not attempt CPR' decision-making: a qualitative study.
The community matron (CM) is often the key worker caring for patients with chronic, life-limiting, long-term conditions, but these patients are not always recognised as palliative cases. This study explored the experiences of CMs with regard to advance care planning (ACP) and 'do not attempt cardiopulmonary resuscitation' (DNACPR) decision-making to understand whether or not they felt adequately prepared for this aspect of their role, and why. Qualitative data were generated from six CMs using a broad interpretive phenomenological approach. Face-to-face recorded interviews were analysed using template analysis. The study found that although participants faced complex ethical situations around ACP and DNACPR almost on a daily basis, none had received any formal training despite the emphasis on training in national and local guidelines. Participants often struggled to get their patients accepted on to the Gold Standards Framework. The research found variability and complexity of cases to be the main barriers to clear identification of the palliative phase.


Automated classification of eligibility criteria in clinical trials to facilitate patient-trial matching for specific patient populations.
To develop automated classification methods for eligibility criteria in ClinicalTrials.gov to facilitate patient-trial matching for specific populations such as persons living with HIV or pregnant women. We annotated 891 interventional cancer trials from ClinicalTrials.gov based on their eligibility for human immunodeficiency virus (HIV)-positive patients using their eligibility criteria. These annotations were used to develop classifiers based on regular expressions and machine learning (ML). After evaluating classification of cancer trials for eligibility of HIV-positive patients, we sought to evaluate the generalizability of our approach to more general diseases and conditions. We annotated the eligibility criteria for 1570 of the most recent interventional trials from ClinicalTrials.gov for HIV-positive and pregnancy eligibility, and the classifiers were retrained and reevaluated using these data. On the cancer-HIV dataset, the baseline regex model, the bag-of-words ML classifier, and the ML classifier with named entity recognition (NER) achieved macro-averaged F2 scores of 0.77, 0.87, and 0.87, respectively; the addition of NER did not result in a significant performance improvement. On the general dataset, ML + NER achieved macro-averaged F2 scores of 0.91 and 0.85 for HIV and pregnancy, respectively. The eligibility status of specific patient populations, such as persons living with HIV and pregnant women, for clinical trials is of interest to both patients and clinicians. We show that it is feasible to develop a high-performing, automated trial classification system for eligibility status that can be integrated into consumer-facing search engines as well as patient-trial matching systems.


Polarization insensitive all-optical up-conversion for ROF systems based on parallel pump FWM in a SOA.
We have proposed and experimentally investigated polarization insensitive all-optical up-conversion for ROF system based on FWM in a semiconductor optical amplifier (SOA). The parallel pump is generated based on odd-order optical sidebands and carrier suppression using an external intensity modulator and a cascaded optical filter. Therefore, the two pumps are always parallel and phase locked, which makes system polarization insensitive. After FWM in a SOA and optical filtering, similar to single sideband (SSB) 40 GHz optical millimeter-wave is generated only using 10 GHz RF as local oscillator (LO). The receiver sensitivity at a BER of 10(-9) for the up-converted signals is -28.4 dBm. The power penalty for the up-converted downstream signals is smaller than 1 dBm after 20 km SSMF-28 transmission.


Shoot-to-Root Signal Transmission Regulates Root Fe(III) Reductase Activity in the dgl Mutant of Pea.
To understand the root, shoot, and Fe-nutritional factors that regulate root Fe-acquisition processes in dicotyledonous plants, Fe(III) reduction and net proton efflux were quantified in root systems of an Fe-hyperaccumulating mutant (dgl) and a parental (cv Dippes Gelbe Viktoria [DGV]) genotype of pea (Pisum sativum). Plants were grown with (+Fe treated) or without (-Fe treated) added Fe(III)-N,N'-ethylenebis[2-(2-hydroxyphenyl)-glycine] (2 [mu]M); root Fe(III) reduction was measured in solutions containing growth nutrients, 0.1 mM Fe(III)-ethylenediaminetetraacetic acid, and 0.1 mM Na2-bathophenanthrolinedisulfonic acid. Daily measurements of Fe(III) reduction (d 10-20) revealed initially low rates in +Fe-treated and -Fe-treated dgl, followed by a nearly 5-fold stimulation in rates by d 15 for both growth types. In DGV, root Fe(III) reductase activity increased only minimally by d 20 in +Fe-treated plants and about 3-fold in -Fe-treated plants, beginning on d 15. Net proton efflux was enhanced in roots of -Fe-treated DGV and both dgl growth types, relative to +Fe-treated DGV. In dgl, the enhanced proton efflux occurred prior to the increase in root Fe(III) reductase activity. Reductase studies using plants with reciprocal shoot:root grafts demonstrated that shoot expression of the dgl gene leads to the generation of a transmissible signal that enhances Fe(III) reductase activity in roots. The dgl gene product may alter or interfere with a normal component of a signal transduction mechanism regulating Fe homeostasis in plants.


Methodological considerations for a randomised controlled trial of podiatry care in rheumatoid arthritis: lessons from an exploratory trial.
Whilst evidence exists to support the use of single treatments such as orthoses and footwear, the effectiveness of podiatry-led care as a complex intervention for patients with rheumatoid arthritis (RA) related foot problems is unknown. The aim of this study was to undertake an exploratory randomised controlled parallel arm clinical trial (RheumAFooT) to inform the design and implementation of a definitive trial and to understand the potential benefits of this care. Patients with a definite diagnosis of RA, stable drug management 3 months prior to entry, and a current history of foot problems (pain, deformity, stiffness, skin or nail lesions, or footwear problems) were recruited from a hospital outpatient rheumatology clinic and randomised to receive 12 months of podiatry treatment or no care. The primary outcome was change in foot health status using the impairment/footwear (LFISIF) and activity limitation/participation restriction (LFISAP) subscales of the Leeds Foot Impact Scale. Disease Activity Score (DAS), Health Assessment Questionnaire (HAQ) score and walking speed (m/s) were also recorded. Of the 80 patients identified, 64 patients were eligible to participate in the pilot and 34 were recruited. 16 patients were randomised to receive podiatry led foot care and 18 received no care. Against a backdrop of stable disease (DAS and HAQ scores), there was a statistically significant between group difference in the change in foot health status for foot impairment (LFISIF) but not activity/participation (LFISAP) or function (walking speed) over 12 months. In the podiatry arm, 1 patient declined treatment following randomisation (did not want additional hospital visits) and 3 self-withdrew (lost to follow-up). Patients received an average of 3 consultations for assessment and treatment comprising routine care for skin and nail lesions (n = 3), foot orthoses (n = 9), footwear referral to the orthotist (n = 5), and ultrasound guided intra-articular steroid injection (n = 1). In this exploratory trial patients were difficult to recruit (stable drug management and co-morbid disease) and retain (lack of benefit/additional treatment burden) but overall the intervention was safe (no adverse reactions). Twelve months of podiatry care maintained but did not improve foot health status. These observations are important for the design and implementation of a definitive randomised controlled trial. ISRCTN: 01982076.


Ribonucleolytic resection is required for repair of strand displaced nonhomologous end-joining intermediates.
Nonhomologous end-joining (NHEJ) pathways repair DNA double-strand breaks (DSBs) in eukaryotes and many prokaryotes, although it is not reported to operate in the third domain of life, archaea. Here, we describe a complete NHEJ complex, consisting of DNA ligase (Lig), polymerase (Pol), phosphoesterase (PE), and Ku from a mesophillic archaeon, Methanocella paludicola (Mpa). Mpa Lig has limited DNA nick-sealing activity but is efficient in ligating nicks containing a 3' ribonucleotide. Mpa Pol preferentially incorporates nucleoside triphosphates onto a DNA primer strand, filling DNA gaps in annealed breaks. Mpa PE sequentially removes 3' phosphates and ribonucleotides from primer strands, leaving a ligatable terminal 3' monoribonucleotide. These proteins, together with the DNA end-binding protein Ku, form a functional NHEJ break-repair apparatus that is highly homologous to the bacterial complex. Although the major roles of Pol and Lig in break repair have been reported, PE's function in NHEJ has remained obscure. We establish that PE is required for ribonucleolytic resection of RNA intermediates at annealed DSBs. Polymerase-catalyzed strand-displacement synthesis on DNA gaps can result in the formation of nonligatable NHEJ intermediates. The function of PE in NHEJ repair is to detect and remove inappropriately incorporated ribonucleotides or phosphates from 3' ends of annealed DSBs to configure the termini for ligation. Thus, PE prevents the accumulation of abortive genotoxic DNA intermediates arising from strand displacement synthesis that otherwise would be refractory to repair.


Recurrent single echo beats after cryoablation of atrioventricular nodal reentrant tachycardia: The pediatric population.
Cryoablation for atrioventricular nodal reentrant tachycardia (AVNRT) is effective and safe with a reported limitation of lower success and higher recurrence rates. We have observed cases in which slow pathway conduction was eliminated as demonstrated by atrial extra-stimulus testing within 1 min of cryo-energy delivery but returned following tissue rewarming. Frequently, slow pathway conduction persisted despite multiple acutely successful lesions over a broad anatomic region. We aimed to determine if return of slow pathway conduction after elimination during cryoablation represents a risk for recurrent AVNRT with the same intermediate term results as slow pathway ablation. We hypothesize that remnant single echo beats in the absence of sustained slow pathway conduction and inducible AVNRT is an acceptable end point after clear slow pathway elimination during cryoablation. Retrospective chart review of patients undergoing attempted slow pathway ablation for AVNRT using solely cryoablation between January 2015-January 2018. Forty-four patients met inclusion criteria with at-least 2 features of dual AVN physiology. 19 patients had return of slow pathway conduction shortly after clear elimination during cryoablation (Group A) while 25 did not (Group B). All in Group A had recurrent single echo beats but none had sustained slow pathway conduction at the end of the procedure nor AVNRT recurrence at 1 year. Recurrent single echo beats with absent sustained slow pathway conduction and non-inducible AVNRT may be an acceptable endpoint for slow pathway ablation of AVNRT using cryoablation when there is elimination of slow pathway demonstrated during energy delivery.


Provider-Initiated HIV Testing for Migrants in Spain: A Qualitative Study with Health Care Workers and Foreign-Born Sexual Minorities.
Provider-initiated HIV testing (PITC) is increasingly adopted in Europe. The success of the approach at identifying new HIV cases relies on its effectiveness at testing individuals most at risk. However, its suitability to reach populations facing overlapping vulnerabilities is under researched. This qualitative study examined HIV testing experiences and perceptions amongst Latin-American migrant men who have sex with men and transgender females in Spain, as well as health professionals' experiences offering HIV tests to migrants in Barcelona and Madrid. We conducted 32 in-depth interviews and 8 discussion groups with 38 Latin-American migrants and 21 health professionals. We imported verbatim transcripts and detailed field work notes into the qualitative software package Nvivo-10 and applied to all data a coding framework to examine systematically different HIV testing dimensions and modalities. The dimensions analysed were based on the World Health Organization "5 Cs" principles: Consent, Counselling, Connection to treatment, Correctness of results and Confidentiality. Health professionals reported that PITC was conceptually acceptable for them, although their perceived inability to adequately communicate HIV+ results and resulting bottle necks in the flow of care were recurrent concerns. Endorsement and adherence to the principles underpinning the rights-based response to HIV varied widely across health settings. The offer of an HIV test during routine consultations was generally appreciated by users as a way of avoiding the embarrassment of asking for it. Several participants deemed compulsory testing as acceptable on public health grounds. In spite of--and sometimes because of--partial endorsement of rights-based approaches, PITC was acceptable in a population with high levels of internalised stigma. PITC is a promising approach to reach sexual minority migrants who hold high levels of internalised stigma but explicit extra efforts are needed to safeguard the rights of the most vulnerable.


Procyanidin content and variation in some commonly consumed foods.
Procyanidins are a subclass of flavonoids found in commonly consumed foods that have attracted increasing attention due to their potential health benefits. However, little is known regarding their dietary intake levels because detailed quantitative information on the procyanidin profiles present in many food products is lacking. Therefore, the procyanidin content of red wine, chocolate, cranberry juice and four varieties of apples has been determined. On average, chocolate and apples contained the largest procyanidin content per serving (164.7 and 147.1 mg, respectively) compared with red wine and cranberry juice (22.0 and 31.9 mg, respectively). However, the procyanidin content varied greatly between apple samples (12.3-252.4 mg/serving) with the highest amounts on average observed for the Red Delicious (207.7 mg/serving) and Granny Smith (183.3 mg/serving) varieties and the lowest amounts in the Golden Delicious (92.5 mg/serving) and McIntosh (105.0 mg/serving) varieties. The compositional data reported herein are important for the initial understanding of which foods contribute most to the dietary intake of procyanidins and may be used to compile a database necessary to infer epidemiological relationships to health and disease.


Social Rejection and Alcohol Use in Daily Life.
Prior studies have found that social rejection is associated with increases in negative affect, distress, and hostility. Fewer studies, however, have examined the impact of social rejection on alcohol use, and no known studies have tested whether the impact of social rejection by close others differs from social rejection by acquaintances in its association with subsequent drinking. Participants completed event-contingent reports of their social interactions and alcohol use for 14 consecutive days on smartphones. Multilevel negative binomial regression models tested whether experiencing more social rejection than usual was associated with increased drinking, and whether this association was stronger when participants were rejected by close others (e.g., friends, spouses, family members) versus strangers or acquaintances. Results showed a significant interaction between social rejection and relationship closeness. On days characterized by rejection by close others, the likelihood of drinking significantly increased. On days characterized by rejection by acquaintances, by contrast, there was no increase in the likelihood of drinking. There was no main effect of rejection on likelihood of drinking. These results suggest that relationship type is a key factor in whether social rejection translates to potentially harmful behaviors, such as increased alcohol use. This finding is in contrast to many laboratory paradigms of rejection, which emphasize rejection and ostracism by strangers rather than known others. In the more naturalistic setting of measuring social interactions on smartphone in daily life, our findings suggest that only social rejection delivered by close others, and not strangers, led to subsequent drinking.


cAMP-signaling pathway acts in selective synergism with glucose or tolbutamide to increase cytosolic Ca2+ in rat pancreatic beta-cells.
cAMP and the insulinotropic peptides that raise cAMP glucose-dependently increase the cytosolic free Ca2+ concentration ([Ca2+]i) in pancreatic beta-cells, which is tightly linked to the potentiation of glucose-induced insulin release. We examined whether cAMP increases [Ca2+]i in specific cooperation only with glucose or also with other insulin secretagogues that act through different mechanisms. [Ca2+]i in single rat pancreatic beta-cells was measured by dual-wavelength fura-2 microfluorometry. In the presence of a stimulatory concentration of glucose (8.3 mmol/l) and the moderate elevation in [Ca2+]i induced by it, forskolin, an activator of adenylyl cyclase, or dibutyryl cAMP produced a marked additional increase in [Ca2+]i but was ineffective at the basal 2.8 mmol/l glucose. These cAMP-elevating agents also potentiated the effect of tolbutamide on [Ca2+]i. The cAMP-induced increase in [Ca2+]i was completely and selectively inhibited by a blocker of cAMP-dependent protein kinase A (PKA), and by nitrendipine, a blocker of the L-type Ca2+ channel. However, in the presence of high KCl and the [Ca2+]i elevation induced by it, a rise in cAMP failed to further increase [Ca2+]i, whereas BAY K8644, an agonist of L-type Ca2+ channels, evoked an additional increase in [Ca2+]i. Under low Na+ conditions, the [Ca2+]i response to cAMP was observed in the majority of the cells. In the cells in which glucose at 4.5-5 mmol/l was inadequate to increase [Ca2+]i, the glucose together with a rise in cAMP often increased [Ca2+]i. Likewise, tolbutamide and a rise in cAMP acted in concert to increase [Ca2+]i. Thus, cAMP left-shifted the concentration-[Ca2+]i response relationship for glucose and tolbutamide. In conclusion, the cAMP-PKA pathway acts in selective synergism with glucose and tolbutamide to initiate [Ca2+]i signals in pancreatic beta- cells. cAMP appears to regulate beta-cell sensitivity to glucose and tolbutamide. In contrast, cAMP fails to cooperate with high KCl to increase [Ca2+]i. It is suggested that cAMP acts mainly on a site that is more proximal but functionally linked to the L-type Ca2+ channel, thereby finally increasing Ca2+ influx through this channel.


Interface strain-induced multiferroicity in a SmFeO3 film.
An epitaxial pseudocubic SmFeO3 thin film on (100) Nb-SrTiO3 was studied based on ferroelectric (FE) characterization and magnetic measurements. High-resolution transmission electron microscopy images clarify the nature of the epitaxial growth, the stress-induced structural distortion at the film/substrate interface, and the existence of two different orientation lattices. Clear grain boundaries can be seen, which could introduce an extra local distortion. Rectangular FE loops can be observed at room temperature, even by just applying a small voltage ranging from -1 to +1 V, indicative of the presence of FE polarization. Piezoelectric force microscopy images confirm the existence of FE domains and the switchable polarization. A strong ferromagnetic-like transition occurs around 185 K, which is much lower than the transition observed in the bulk sample. It is believed that the pseudocubic structure enhances FE polarization and decreases the magnetic ordering temperature, which is confirmed by the first-principles theoretical calculations. Meanwhile, the ferroelectricity in this thin film should originate from distortion and modification in the structural modules rather than from the exchange striction interaction that is found in the bulk SmFeO3.


Evolutionary Cell Biology of Proteins from Protists to Humans and Plants.
During evolution, the cell as a fine-tuned machine had to undergo permanent adjustments to match changes in its environment, while "closed for repair work" was not possible. Evolution from protists (protozoa and unicellular algae) to multicellular organisms may have occurred in basically two lineages, Unikonta and Bikonta, culminating in mammals and angiosperms (flowering plants), respectively. Unicellular models for unikont evolution are myxamoebae (Dictyostelium) and increasingly also choanoflagellates, whereas for bikonts, ciliates are preferred models. Information accumulating from combined molecular database search and experimental verification allows new insights into evolutionary diversification and maintenance of genes/proteins from protozoa on, eventually with orthologs in bacteria. However, proteins have rarely been followed up systematically for maintenance or change of function or intracellular localization, acquirement of new domains, partial deletion (e.g. of subunits), and refunctionalization, etc. These aspects are discussed in this review, envisaging "evolutionary cell biology." Protozoan heritage is found for most important cellular structures and functions up to humans and flowering plants. Examples discussed include refunctionalization of voltage-dependent Ca2+ channels in cilia and replacement by other types during evolution. Altogether components serving Ca2+ signaling are very flexible throughout evolution, calmodulin being a most conservative example, in contrast to calcineurin whose catalytic subunit is lost in plants, whereas both subunits are maintained up to mammals for complex functions (immune defense and learning). Domain structure of R-type SNAREs differs in mono- and bikonta, as do Ca2+ -dependent protein kinases. Unprecedented selective expansion of the subunit a which connects multimeric base piece and head parts (V0, V1) of H+ -ATPase/pump may well reflect the intriguing vesicle trafficking system in ciliates, specifically in Paramecium. One of the most flexible proteins is centrin when its intracellular localization and function throughout evolution is traced. There are many more examples documenting evolutionary flexibility of translation products depending on requirements and potential for implantation within the actual cellular context at different levels of evolution. From estimates of gene and protein numbers per organism, it appears that much of the basic inventory of protozoan precursors could be transmitted to highest eukaryotic levels, with some losses and also with important additional "inventions."


Outcomes and outcome measures used in evaluation of communication training in oncology - a systematic literature review, an expert workshop, and recommendations for future research.
Communication between health care provider and patients in oncology presents challenges. Communication skills training have been frequently developed to address those. Given the complexity of communication training, the choice of outcomes and outcome measures to assess its effectiveness is important. The aim of this paper is to 1) perform a systematic review on outcomes and outcome measures used in evaluations of communication training, 2) discuss specific challenges and 3) provide recommendations for the selection of outcomes in future studies. To identify studies and reviews reporting on the evaluation of communication training for health care professionals in oncology, we searched seven databases (Ovid MEDLINE, CENTRAL, CINAHL, EMBASE, PsychINFO, PsychARTICLES and Web of Science). We extracted outcomes assessed and the respective assessment methods. We held a two-day workshop with experts (n = 16) in communication theory, development and evaluation of generic or cancer-specific communication training and/or outcome measure development to identify and address challenges in the evaluation of communication training in oncology. After the workshop, participants contributed to the development of recommendations addressing those challenges. Out of 2181 references, we included 96 publications (33 RCTs, 2 RCT protocols, 4 controlled trials, 36 uncontrolled studies, 21 reviews) in the review. Most frequently used outcomes were participants' training evaluation, their communication confidence, observed communication skills and patients' overall satisfaction and anxiety. Outcomes were assessed using questionnaires for participants (57.3%), patients (36.0%) and observations of real (34.7%) and simulated (30.7%) patient encounters. Outcomes and outcome measures varied widely across studies. Experts agreed that outcomes need to be precisely defined and linked with explicit learning objectives of the training. Furthermore, outcomes should be assessed as broadly as possible on different levels (health care professional, patient and interaction level). Measuring the effects of training programmes aimed at improving health care professionals' communication skills presents considerable challenges. Outcomes as well as outcome measures differ widely across studies. We recommended to link outcome assessment to specific learning objectives and to assess outcomes as broadly as possible.


Theory-of-mind in adolescents and young adults with Alström syndrome.
The study focuses on theory-of-mind in adolescents and young adults with Alström syndrome (ALMS). ALMS, an autosomal recessive syndrome causes juvenile blindness, sensorineural hearing loss, cardiomyopathy, endocrinological disorders and metabolic dysfunction. Theory-of-mind (ToM) refers to the ability to impute mental states to one self and to others. Clinical observations have revealed an increased occurrence of deviances in mental state understanding in ALMS. In the present study ToM will be examined and related to working memory (WM), verbal ability and sensory loss. Twelve young individuals (16-37 years) with ALMS and 24 nondisabled individuals matched on age, gender and educational level participated. ToM was assessed by means of a multiple task that taxes the ability to understand thoughts and feelings of story characters'. WM was examined by means of a reading span task and verbal ability by means of a vocabulary test. The ALMS group performed at significantly lower levels in ToM tasks and displayed a higher variability in performance than the control group. Individuals with ALMS and a relatively poor level performance provided fewer correct mental state inferences in ToM tasks than ALMS individuals with relatively higher performance levels. ALMS individuals with relatively high performance levels made as many correct inferences in ToM tasks as the control group, but their inferences were more often incomplete. Vocabulary skills and educational level, but not WM-capacity predicted ToM performance. Degree of deafblindness did not have an impact on ToM. Age of onset of visual loss but not hearing loss related to ToM. The individuals with ALMS display a high degree of heterogeneity in terms of ToM, where some individuals reached performance levels comparable to nondisabled individuals. The results are discussed with respect to how cognitive and verbal abilities and factors related to the disability affect ToM.


Astrocyte and Neuronal Plasticity in the Somatosensory System.
Changing the whisker complement on a rodent's snout can lead to two forms of experience-dependent plasticity (EDP) in the neurons of the barrel cortex, where whiskers are somatotopically represented. One form, termed coding plasticity, concerns changes in synaptic transmission and connectivity between neurons. This is thought to underlie learning and memory processes and so adaptation to a changing environment. The second, called homeostatic plasticity, serves to maintain a restricted dynamic range of neuronal activity thus preventing its saturation or total downregulation. Current explanatory models of cortical EDP are almost exclusively neurocentric. However, in recent years, increasing evidence has emerged on the role of astrocytes in brain function, including plasticity. Indeed, astrocytes appear as necessary partners of neurons at the core of the mechanisms of coding and homeostatic plasticity recorded in neurons. In addition to neuronal plasticity, several different forms of astrocytic plasticity have recently been discovered. They extend from changes in receptor expression and dynamic changes in morphology to alteration in gliotransmitter release. It is however unclear how astrocytic plasticity contributes to the neuronal EDP. Here, we review the known and possible roles for astrocytes in the barrel cortex, including its plasticity.


Default-mode activity during a passive sensory task: uncoupled from deactivation but impacting activation.
Deactivation refers to increased neural activity during low-demand tasks or rest compared with high-demand tasks. Several groups have reported that a particular set of brain regions, including the posterior cingulate cortex and the medial prefrontal cortex, among others, is consistently deactivated. Taken together, these typically deactivated brain regions appear to constitute a default-mode network of brain activity that predominates in the absence of a demanding external task. Examining a passive, block-design sensory task with a standard deactivation analysis (rest epochs vs. stimulus epochs), we demonstrate that the default-mode network is undetectable in one run and only partially detectable in a second run. Using independent component analysis, however, we were able to detect the full default-mode network in both runs and to demonstrate that, in the majority of subjects, it persisted across both rest and stimulus epochs, uncoupled from the task waveform, and so mostly undetectable as deactivation. We also replicate an earlier finding that the default-mode network includes the hippocampus suggesting that episodic memory is incorporated in default-mode cognitive processing. Furthermore, we show that the more a subject's default-mode activity was correlated with the rest epochs (and "deactivated" during stimulus epochs), the greater that subject's activation to the visual and auditory stimuli. We conclude that activity in the default-mode network may persist through both experimental and rest epochs if the experiment is not sufficiently challenging. Time-series analysis of default-mode activity provides a measure of the degree to which a task engages a subject and whether it is sufficient to interrupt the processes--presumably cognitive, internally generated, and involving episodic memory--mediated by the default-mode network.


Inexpensive multigram-scale synthesis of cyclic enamines and 3-N spirocyclopropyl systems.
Cyclic enamines are important synthons for many synthetic and pharmacological targets. Here, we report an inexpensive, catalyst-free, multigram-scale synthesis for cyclic enamines with exocyclic double bonds and four- to seven-membered rings. This strategy is more conducive to scale up, permissive of functionalization around the cyclic system, and less sensitive to the nature of the N-protecting group than previously-described methods for cyclic enamine synthesis. Further, we explore application of these enamines to the synthesis of highly-strained spirocyclic 3N-cyclopropyl scaffolds.


Infant learning is influenced by local spurious generalizations.
In previous work, 11-month-old infants were able to learn rules about the relation of the consonants in CVCV words from just four examples. The rules involved phonetic feature relations (same voicing or same place of articulation), and infants' learning was impeded when pairs of words allowed alternative possible generalizations (e.g. two words both contained the specific consonants p and t). Experiment 1 asked whether a small number of such spurious generalizations found in a randomly ordered list of 24 different words would also impede learning. It did - infants showed no sign of learning the rule. To ask whether it was the overall set of words or their order that prevented learning, Experiment 2 reordered the words to avoid local spurious generalizations. Infants showed robust learning. Infants thus appear to entertain spurious generalizations based on small, local subsets of stimuli. The results support a characterization of infants as incremental rather than batch learners.


Mobilisation strategies after hip fracture surgery in adults.
Hip fracture mainly occurs in older people. Mobilisation strategies such as gait retraining and exercises are used at various stages of rehabilitation after surgery. To evaluate the effects of different mobilisation strategies after hip fracture surgery in adults. We searched the Cochrane Bone, Joint and Muscle Trauma Group Specialised Register, the Cochrane Central Register of Controlled Trials, MEDLINE and other databases, conference proceedings and reference lists of articles, up to January 2006. All randomised or quasi-randomised trials comparing different mobilisation strategies after hip fracture surgery. The authors independently selected trials, assessed trial quality and extracted data. There was no data pooling. Most of the 13 included trials (involving 1065 participants, generally over 65 years) were small and all had methodological limitations, including inadequate follow up. Seven trials evaluated mobilisation strategies started soon after hip fracture surgery. One historic trial found no significant differences in unfavourable outcomes for weight bearing started at two versus 12 weeks after internal fixation of a displaced intracapsular fracture. Two trials compared a more with a less intensive regimen of physiotherapy: one found no difference in recovery, the other found a higher level of drop-out in the more intensive group with no difference in length of hospital stay. One trial found short-term improvement in mobility and balance for a two-week programme of weight-bearing versus non-weight-bearing exercise. One trial found improved mobility in those given a quadriceps muscle strengthening exercise programme. One trial found no significant difference in recovery of mobility after a treadmill versus conventional gait retraining programme. One trial found a greater recovery of pre-fracture mobility after neuromuscular stimulation of the quadriceps muscle. Six trials evaluated strategies started after hospital discharge. Started soon after discharge, two trials found improved outcome after 12 weeks of intensive physical training and a home-based physical therapy programme respectively. Begun after completion of standard physical therapy, one trial found improved outcome after six months of intensive physical training whereas another trial found no significant effects of home-based resistance or aerobic training. One trial found improved outcome after home-based exercises started around 22 weeks from injury. One trial found home-based weight-bearing exercises starting at seven months produced no statistically significant differences aside for greater quadriceps strength. There is insufficient evidence from randomised trials to establish the effectiveness of the various mobilisation strategies used in rehabilitation after hip fracture surgery. Further research is required to establish the possible benefits of the additional provision of interventions, including intensive supervised exercises, primarily aimed at enhancing mobility.


Examining Primary Care Health Encounters for Adults With Autism Spectrum Disorder.
Our objective was to identify perceived barriers and strategies to improve primary care encounters, as reported by adults with autism spectrum disorder (ASD), caregivers of adults with ASD, and primary care providers (PCPs) treating adults with ASD. As part of a larger mixed-methods design, adults with ASD, caregivers, and PCPs (N = 78) in Los Angeles and Philadelphia completed surveys examining barriers to care and strategies to improve care. Multiple barriers to care were reported by adults with ASD and caregivers, including communication and sensory challenges. Adults with ASD and caregivers reported minimal use of strategies during primary care visits but indicated that those used were helpful during care. Expert PCPs reported using strategies more frequently than novice PCPs. All respondent groups endorsed that strategies had the potential to improve care in the future for adults with ASD. Opportunities exist for occupational therapy collaboration in primary health care and primary care education to improve care for adults with ASD.


Do Intervention Strategies of Women Healthy Heart Project (WHHP) Impact on Differently on Working and Housewives?
The purpose of this study is to evaluate the possible difference of the impact of Women Healthy Heart Project on lifestyle, as well as physical/biochemical parameters of working women and housewives. This was a community-based intervention study conducted over 5 years (2002-2007) in the counties of Isfahan and Najafabad (intervention areas) and Arak (control). Pre-study situation analysis of women was followed by 5 years of wide-ranging interventions (educational/environmental) conducted by various organizations using different methodologies. The interventions were aimed at modifying/improving lifestyle by increasing physical activity, encouraging healthy eating, and tobacco use cessation. The organizations involved in performing the interventions included the local radio and television authority, health/treatment centers, the Red Crescent Society, Municipalities, the Relief Committee, the Center for Retirees' Welfare, and the Literacy Campaign Organization. After 5 years, final phase same as first phase was planed. The subjects studied in all phases' the pre- and post-intervention phases consisted of 10586 women aged above 18 years. Demographic data, obesity index, smoking, physical activity and eating habit were assessed before and after the study. Data were analyzed using SPSS-15 using Student's t-test, chi-square test, the general linear model of ANOVA, and logistic regression. We studied 10586 women (6105 and 4481 women, pre- and post-intervention, respectively). Mean age of working women was 34.14±10.09 and 34.08±9.35 years before and after the study, respectively. Mean age of housewives before and after the study was 40.05±14.61 and 40.36±15.32 years, respectively. Interventions conducted during 5 years improved eating habits and decreased tobacco use in working women and housewives. In every phase of the study, there was a significant age difference between housewives and working women (P<0.001). Hence data were adjusted according to age in each phase. Overall physical activity of housewives and working women increased, but the percentage of passive smokers among housewives remained unchanged. Waist circumference and waist-to-hip ratio changed significantly in housewives following interventions (P<0.001). The parameter which improved significantly in working women was waist circumference (P<0.05). However, after adjusting for age, no significant difference was seen between working women and housewives following interventions. Community-based interventions, especially those directed at housewives, can lead to significant improvements in lifestyle and cardiovascular risk reduction. It seems that working women require tailored interventions to suit their conditions. Although short-term outcomes may seem insignificant, persistence and continuity of lifestyle changes may lead to reduced prevalence of cardiovascular diseases. Then longer-term studies are warranted.


A rare periosteal diaphyseal lesion of the ulna.
Periosteal lesions of the ulna diaphysis are rare, include a wide spectrum of tumors, and may cause considerable diagnostic problems. Surgical treatment may vary widely, based on an accurate diagnosis. We present the case of a periosteal, extraskeletal low grade myxoid chondrosarcoma of the ulna diaphysis. The surgical therapy included an en-bloc resection with allograft reconstruction. The patient showed a favorable outcome. Careful preoperative evaluation and planning are imperative to obtain a satisfactory oncological and functional outcome, especially with uncommon tumor presentations at rare locations.


Geographic distribution of Pythium insidiosum infections in the United States.
To describe the geographic distribution of infections caused by Pythium insidiosum in dogs, horses, and other animal species in the US. For the last 20 years, we have collected data from cases of pythiosis in 1,150 horses, 467 dogs, and other species (59) from various geographic locations in the US. Due to lost data (from 2006 to 2016), the selected cases include years 2000 to 2005 and 2016 to 2020. The selection of cases was based on infected host clinical features, serum samples demonstrating strong positive anti-P insidiosum IgG titers in serologic assays, and positive results on ≥ 1 of the following diagnostic modalities: microbial culture on 2% Sabouraud dextrose agar, histologic evaluation, PCR assay, and wet mount cytologic evaluation (with potassium hydroxide). Most confirmed P insidiosum infections were found in horses and dogs in the southeastern US. Interestingly, in Texas, no cases were found west of longitude 100°W. Few pythiosis cases were diagnosed in west-coast states. Equine cases were more often diagnosed during summer and fall months, but canine cases were more often diagnosed between September and February. Cases in other species were discovered in the same geographic areas as those in dogs and horses. To our knowledge, this is the first report providing the ecological distribution of P insidiosum infection in affected species in the US. Results of this study illustrated the importance of including P insidiosum in the differential diagnostic scheme of nonhealing skin lesions or intestinal granulomatous masses, particularly in dogs and horses inhabiting or having visited endemic areas.


Antioxidant Activity of Hemp (Cannabis sativa L.) Seed Oil in Drosophila melanogaster Larvae under Non-Stress and H2O2-Induced Oxidative Stress Conditions.
The oil extracted from hemp seeds has significant nutritional and biological properties due to the unique composition of polyunsaturated fatty acids and various antioxidant compounds. The potential of this oil for the prevention of oxidative stress and for the treatment of oxidative-stress-induced ailments is of increasing interest. Most studies of hemp seed oil were conducted in-vitro, meaning we lack information about effects and activity in vivo. In the present study, we evaluated the hypothesis that hemp seed oil at different concentrations improves the oxidative state of D. melanogaster, under non-stress as well as hydrogen-peroxide-induced stress. We analyzed the effects of hemp seed oil on oxidative stress markers and on the life cycle of D.melanogaster under non-stress and hydrogen-peroxide-induced stress conditions. D.melanogaster larvae were exposed to hemp seed oil concentrations ranging from 12.5 to 125 μL/mL. The results revealed that under non-stress conditions, oil concentrations up to 62.5 µL/mL did not induce negative effects on the life cycle of D. melanogaster and maintained the redox status of the larval cells at similar levels to the control level. Under oxidative stress conditions, biochemical parameters were significantly affected and only two oil concentrations, 18.7 and 31.2 µL/mL, provided protection against hydrogen peroxide stress effects. A higher oil concentration (125 μL/mL) exerted negative effects on the oxidative status and increased larval mortality. The tested oil was characterized chemically by NMR, transesterification, and silylation, followed by GC-MS analyses, and was shown to contain polyunsaturated fatty acid triglycerides and low levels of tocopherols. The high levels of linoleic and linolenic acids in the oil are suggested to be responsible for the observed in vivo antioxidant effects. Taken together, the results show that hemp seed oil is effective for reducing oxidative stress at the cellular level, thus supporting the hypothesis. The obtained results point to the potential of hemp seed oil for the prevention and treatment of conditions caused by the action of reactive oxygen species.


Nondissociating cationic immune complexes can deposit in glomerular basement membrane.
The mechanisms by which immune complexes deposit in the glomerular basement membrane have been the subject of much debate, with the relative importance of direct deposition of circulating immune complexes (IC) vs. formation of IC in situ from the binding of circulating antibody to structural or exogenous planted antigen being at issue. In order to determine whether intact IC can deposit as such, covalently linked IC were prepared by a two-step reaction involving the bifunctional reagent toluene-2,4-diisocyanate (TDI), the antigen bovine gamma globulin (BGG), and rabbit anti-BGG antibody. Antigen and antibody were covalently cross-linked, with little self-linkage of antigen or antibody, and IC were purified by gel filtration. The net charge of the complexes was varied by chemical means, either before or after IC formation. When cationic IC were injected intravenously into mice, there was codeposition of antigen and antibody diffusely in the glomerular basement membrane (GBM), and deposits were observed ultrastructurally in the laminae rarae, interna and externa, and the lamina densa. Thus, under conditions of restricted appropriate charge, intact IC can cross the glomerular basement membrane and deposit in subepithelial sites without being excluded by size alone.


The effectiveness of Internet-based blood glucose monitoring system on improving diabetes control in adolescents with type 1 diabetes.
To determine whether the use of an Internet-based blood glucose monitoring system could improve glycemic control in adolescents with type 1 diabetes mellitus (T1DM). In a randomized, controlled clinical trial, a total of 70 adolescent subjects with T1DM were recruited. Subjects randomized to the intervention group (n = 36) were instructed to submit their blood glucose levels weekly by Internet to the diabetes care team during a period of 6 months. Subjects randomized to the control group (n = 34) did not submit results but were under routine follow-up. At baseline, patients were 15.1 ± 2.6 years of age with mean HbA1c of 8.3 ± 1.3%. At the 6-month follow-up period, no by-group differences in change from baseline to end of treatment HbA1c levels were detected. In the intervention group, 12/36 did not submit blood glucose levels and were classified as non-compliant. In a secondary exploratory analysis in which non-compliant patients were omitted, HbA1c values in the compliant intervention group declined from 8.5 ± 1.7% at baseline to 8.2 ± 1.2% at 6 months, while in the control group HbA1c values increased from 8.2 ± 1.1 to 8.4 ± 1.1%, this difference did not reach statistical significance. An Internet-based blood glucose monitoring system was not associated with improved glycemic control in adolescents with T1DM. Identification of a sub-group of compliant subjects who may improve metabolic control by using this tool is needed.


Origin of HIV-1 in the chimpanzee Pan troglodytes troglodytes.
The human AIDS viruses human immunodeficiency virus type 1 (HIV-1) and type 2 (HIV-2) represent cross-species (zoonotic) infections. Although the primate reservoir of HIV-2 has been clearly identified as the sooty mangabey (Cercocebus atys), the origin of HIV-1 remains uncertain. Viruses related to HIV-1 have been isolated from the common chimpanzee (Pan troglodytes), but only three such SIVcpz infections have been documented, one of which involved a virus so divergent that it might represent a different primate lentiviral lineage. In a search for the HIV-1 reservoir, we have now sequenced the genome of a new SIVcpzstrain (SIVcpzUS) and have determined, by mitochondrial DNA analysis, the subspecies identity of all known SIVcpz-infected chimpanzees. We find that two chimpanzee subspecies in Africa, the central P. t. troglodytes and the eastern P. t. schweinfurthii, harbour SIVcpz and that their respective viruses form two highly divergent (but subspecies-specific) phylogenetic lineages. All HIV-1 strains known to infect man, including HIV-1 groups M, N and O, are closely related to just one of these SIVcpz lineages, that found in P. t. troglodytes. Moreover, we find that HIV-1 group N is a mosaic of SIVcpzUS- and HIV-1-related sequences, indicating an ancestral recombination event in a chimpanzee host. These results, together with the observation that the natural range of P. t. troglodytes coincides uniquely with areas of HIV-1 group M, N and O endemicity, indicate that P. t. troglodytes is the primary reservoir for HIV-1 and has been the source of at least three independent introductions of SIVcpz into the human population.


Treatment of carotid artery aneurysms with platinum coils: an experimental study in dogs.
The objectives of this study were to investigate the consistency, patency, and natural history of a vein graft canine aneurysm model and to determine the effectiveness of various coil designs on inducing aneurysm thrombosis. Twenty-one sacculuslike aneurysms were created in mongrel dogs by anastomosing a vein pouch to the common carotid artery. The model produced wide-neck aneurysms with 100% patency. The canine vein graft aneurysm provided an excellent model for the evaluation of endovascular devices. Three types of specially designed platinum coils were placed in the aneurysms: those with simple curves, those with complex curves, and those with flower petal curves and silk fibers. These coils were placed by the endovascular route by means of microcatheters. Flower petal coils with silk fibers were effective in producing thrombosis of the aneurysms, suggesting that coils of the appropriate design may be useful in the endovascular treatment of aneurysms. The other coil designs evaluated, those with simple and complex curves without silk fibers, demonstrated insufficient thrombogenicity and spatial stability.


Serological testing versus other strategies for diagnosis of active tuberculosis in India: a cost-effectiveness analysis.
Undiagnosed and misdiagnosed tuberculosis (TB) drives the epidemic in India. Serological (antibody detection) TB tests are not recommended by any agency, but widely used in many countries, including the Indian private sector. The cost and impact of using serology compared with other diagnostic techniques is unknown. Taking a patient cohort conservatively equal to the annual number of serological tests done in India (1.5 million adults suspected of having active TB), we used decision analysis to estimate costs and effectiveness of sputum smear microscopy (US$3.62 for two smears), microscopy plus automated liquid culture (mycobacterium growth indicator tube [MGIT], US$20/test), and serological testing (anda-tb ELISA, US$20/test). Data on test accuracy and costs were obtained from published literature. We adopted the perspective of the Indian TB control sector and an analysis frame of 1 year. Our primary outcome was the incremental cost per disability-adjusted life year (DALY) averted. We performed one-way sensitivity analysis on all model parameters, with multiway sensitivity analysis on variables to which the model was most sensitive. If used instead of sputum microscopy, serology generated an estimated 14,000 more TB diagnoses, but also 121,000 more false-positive diagnoses, 102,000 fewer DALYs averted, and 32,000 more secondary TB cases than microscopy, at approximately four times the incremental cost (US$47.5 million versus US$11.9 million). When added to high-quality sputum smears, MGIT culture was estimated to avert 130,000 incremental DALYs at an incremental cost of US$213 per DALY averted. Serology was dominated by (i.e., more costly and less effective than) MGIT culture and remained less economically favorable than sputum smear or TB culture in one-way and multiway sensitivity analyses. In India, sputum smear microscopy remains the most cost-effective diagnostic test available for active TB; efforts to increase access to quality-assured microscopy should take priority. In areas where high-quality microscopy exists and resources are sufficient, MGIT culture is more cost-effective than serology as an additional diagnostic test for TB. These data informed a recently published World Health Organization policy statement against serological tests.


Five-Year Survival of Short Single-Tooth Implants (6 mm): A Randomized Controlled Clinical Trial.
The aim of the present study was to evaluate whether 6-mm dental implants in the posterior segments of either jaw perform equally well in terms of clinical and radiographic outcomes when compared with 10-mm implants after 5 y of loading. Patients with single-tooth gaps in the posterior area who were scheduled for implant therapy were randomly assigned to a group receiving either a 6- or 10-mm implant. After a healing period of 10 wk, implants were loaded with a screw-retained single crown and followed up at yearly intervals. Of 96 patients, 86 could be recalled after 5 y. The implant survival rates amounted to 91% (95% confidence interval: 0.836 to 0.998) for the 6-mm group and 100% for the 10-mm group ( P = 0.036). Median crown-to-implant (C/I) ratios were 1.75 (interquartile range [IQR], 1.50 to 1.90) for the 6-mm group and 1.04 (IQR, 0.95 to 1.15) for the 10-mm group, whereas the median marginal bone levels measured -0.29 mm (IQR, -0.92 to 0.23) for the 6-mm group and -0.15 mm (IQR: -0.93 - 0.41) for the 10-mm group after 5 y. The C/I ratio turned out to be statistically significant ( P < 0.001), whereas marginal bone levels showed no significant difference between the groups. The 6-mm implants exhibited significantly lower survival rates than the 10-mm implants over 5 y, whereas there was no difference between upper and lower jaws in terms of survival ( P = 0.58). Lost implants did not show any sign of marginal bone loss or peri-implant infection previous to loss of osseointegration. High C/I ratio and implant length had no significant effect on marginal bone level changes or technical and biological complications (German Clinical Trials Registry: DRKS00006290).


Haplotype study of intermediate-length alleles at the fragile X (FMR1) gene: ATL1, FMRb, and microsatellite haplotypes differ from those found in common-size FMR1 alleles.
The CGG repeat within the X-chromosome-linked FMR1 gene, which in hyperexpansion (> 200 copies) results in fragile X syndrome, is highly polymorphic. The mechanism of expansion is not well understood, but CGG repeats called intermediate-length or gray zone alleles (approximately equal 35-60 repeats) are thought to make up the FMR1 alleles showing initial steps in this expansion process. It has been hypothesized that the background haplotype of these alleles plays a role in their susceptibility to expansion. In this study we investigate whether or not the frequencies of alleles and haplotypes at four marker loci in the FMR1 gene region (microsatellites DXS548 and FRAXAC1 and SNPs ATL1 and FMRb) in 84 intermediate-length male chromosomes differ from those in 94 common-size male alleles. The ATL1*G and FMRB*A alleles were more frequent among intermediate-length alleles than among common alleles. In addition, the DXS548-FRAXAC1 T50-T42 and T40-T42 haplotypes were strongly associated with intermediate-length alleles between 41 and 60 CGG repeats (p < 0.001). Two extended haplotypes, DXS548-FRAXAC1-ATL1-FMRb T50-T42-G-A and T40-T42-G-A, are strongly associated (p < 0.001) with intermediate-length alleles between 41 and 60 CGG repeats, and these haplotypes have also been reported as fragile X associated haplotypes in European populations. These data suggest that these haplotypes are among the most susceptible to further expansion among the intermediate-length alleles. T50-T42-G-A was also much more prevalent in males with 35-40 CGG repeats than in males with common-size alleles. ATL1 did not increase discrimination among intermediate-length alleles beyond that detected by DXS548-FRAXAC1 haplotypes, but the FMRb locus did, particularly for the DXS548-FRAXAC1-ATL1 T50-T42-G and T40-T42-G haplotypes. Comparison with fragile X associated haplotypes, from the literature, suggests that repeat hyperexpansion occurs most frequently on chromosomes carrying FMRB*A. Within the intermediate-length allele category, however, there were some significant differences in haplotype frequencies between smaller and larger alleles, and this finding has implications for future studies.


Inhibition of Lysyl Oxidase by Cortisol Regeneration in Human Amnion: Implications for Rupture of Fetal Membranes.
The mechanisms underlying human parturition are still not understood, yet we need this knowledge to combat preterm birth. Fetal membranes express abundant 11β-hydroxysteroid dehydrogenase 1 (11β-HSD1), which converts inert cortisone to active cortisol. We examined whether cortisol regeneration in the amnion might play a role in human parturition through regulation of lysyl oxidase (LOX), a collagen cross-linking enzyme, thereby contributing to the rupture of fetal membranes. By using cultured human primary amnion fibroblasts, we demonstrated that, in addition to the induction of the key enzymes involved in prostaglandin E2 (PGE2) synthesis, cortisol stimulated 11β-HSD1 and inhibited LOX reciprocally. These results were reproduced in human amnion tissue explants after cortisol treatment. Cortisone also inhibited LOX expression, which was abolished by the inhibition of 11β-HSD1. Despite the inhibition of LOX by PGE2, inhibition of the PGE2 pathway failed to block the inhibition of LOX by cortisol. However, inhibition of glucocorticoid receptor and mutation of a negative glucocorticoid response element in LOX promoter abolished the inhibition of LOX by cortisol. Chromatin immunoprecipitation assay revealed that cortisol increased GR binding to the LOX promoter. Moreover, increased cortisol and 11β-HSD1 abundance and decreased LOX abundance were observed in human amnion tissue after the labor-initiated spontaneous rupture of membranes. These data highlight a crucial role for local cortisol regeneration by 11β-HSD1 in the down-regulation of LOX expression via glucocorticoid receptor binding to a negative glucocorticoid response element to its promoter in the amnion, which may contribute to rupture of fetal membranes at parturition.


Ether lipid and sphingolipid expression patterns are estrogen receptor-dependently altered in breast cancer cells.
Identifying co-expression of lipid species is challenging, but indispensable to identify novel therapeutic targets for breast cancer treatment. Lipid metabolism is often dysregulated in cancer cells, and changes in lipid metabolism affect cellular processes such as proliferation, autophagy, and tumor development. In addition to mRNA analysis of sphingolipid metabolizing enzymes, we performed liquid chromatography time-of-flight mass spectrometry analysis in three breast cancer cell lines. These breast cancer cell lines differ in estrogen receptor and G-protein coupled estrogen receptor 1 status. Our data show that sphingolipids and non-sphingolipids are strongly increased in SKBr3 cells. SKBr3 cells are estrogen receptor negative and G-protein coupled estrogen receptor 1 positive. Treatment with G15, a G-protein coupled estrogen receptor 1 antagonist, abolishes the effect of increased sphingolipid and non-sphingolipid levels in SKBr3 cells. In particular, ether lipids are expressed at much higher levels in cancer compared to normal cells and are strongly increased in SKBr3 cells. Our analysis reveals that this is accompanied by increased sphingolipid levels such as ceramide, sphingadiene-ceramide and sphingomyelin. This shows the importance of focusing on more than one lipid class when investigating molecular mechanisms in breast cancer cells. Our analysis allows unbiased screening for different lipid classes leading to identification of co-expression patterns of lipids in the context of breast cancer. Co-expression of different lipid classes could influence tumorigenic potential of breast cancer cells. Identification of co-regulated lipid species is important to achieve improved breast cancer treatment outcome.


Ketoconazole potentiates the antitumor effects of nocodazole: In vivo therapy for human tumor xenografts in nude mice.
Our previous studies demonstrated that the oral antifungal agent ketoconazole (KT) induces apoptosis and G0/G1 phase cell cycle arrest in human cancer cell lines. In this study, we first demonstrated that KT (1 microM) potentiated the apoptotic effects of nocodazole (ND, 1 nM) in COLO 205 cancer cells. We further demonstrated the therapeutic efficacy of a combined treatment of KT (50 mg/kg/three times per week) and ND (5 mg/kg/three times per week) in vivo by treating athymic mice bearing COLO 205 tumor xenografts. The antitumor effects of ND were significantly potentiated by KT in mice after 6 wk of treatment. No gross signs of toxicity were observed in mice receiving these treatment regimens. The apoptotic cells were detected in a microscopic view of the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling staining and by observation of DNA fragmentation in KT + ND-treated tumor tissues. The levels of cell cycle regulatory proteins were determined by Western blot analysis. Treatment with KT inhibits tumor growth through elevation of p53, p21/CIP1, and p27/KIP1 as well as inhibition of cyclin D3 and cyclin-dependent kinase 4 protein expression. Immunohistochemical staining analysis showed that p53, p21/CIP1, and p27/KIP1 immunoreactivity were induced in the tumor tissues. To clarify the roles of the p21/CIP1 and p27/KIP1 protein expression involved in G(0)/G(1) arrest and/or apoptosis induced by a combined treatment with KT and ND, antisense oligodeoxynucleotides (ODNs) specific to p21/CIP1 and p27/KIP1 were used. Our results demonstrated that apoptotic phenomena, including BAX induction and cytochrome C released from mitochondria induced by KT + ND, were significantly attenuated by pretreatment the cells with the p27/KIP1-specific antisense ODNs. These results indicate that p27/KIP1 protein does indeed play a critical role in the KT + ND-induced apoptosis. Our study revealed the molecular mechanism of KT + ND in regression of the tumor growth. The apoptotic effects of KT in a great variety of cancer cells make it a very attractive agent for cancer chemotherapy.


Effects of treatment of rheumatoid arthritis patients with an antibody against tumour necrosis factor alpha on reticuloendothelial and intrapulmonary granulocyte traffic.
The therapeutic effects of a monoclonal antibody against tumour necrosis factor alpha (TNFalpha) were evaluated objectively in 10 patients with rheumatoid arthritis by 111In-labelled granulocyte imaging before and after treatment, and compared with changes in granulocyte kinetics with respect to the liver, spleen and lungs. Anti-TNFalpha resulted in a decrease in the size of the whole-body pool of marginating granulocytes, as reflected by a significant increase in the 30 min intravascular recovery of labelled granulocytes from 40% (S.D. 10) to 47% (S.D. 16) of injected activity (P<0.02). The 111In contents of the spleen, liver and lungs were unchanged, so the origin of the increment in recovery was presumed to be a reduction in granulocyte margination in inflamed synovium, although this was not quantifiable. The sizes of the granulocyte pools in the liver and lungs, expressed as the 111In content of the organ per unit of circulating 111In-labelled cells, were not significantly different after treatment, but the splenic granulocyte pool decreased by 16% (S.D. 19) (P<0.05). Individual changes in the size of the splenic pool showed no significant correlation with corresponding changes in 30 min recovery or with corresponding indices of inflammation (24 h 111In-granulocyte joint activity and C-reactive protein). We conclude that anti-TNFalpha produces an obvious resolution in inflammatory joint activity that is accompanied by an increased circulating component of the total blood granulocyte pool, as a result of decreased margination at sites of inflammation. Anti-TNFalpha may also produce a specific decrease in splenic granulocyte pooling, independent of any anti-inflammatory effects, although a similar decrease in the lungs, which might be anticipated as a result of reduced cytokine-induced granulocyte activation, could not be detected.