Reflectance confocal microscopy for diagnosing cutaneous melanoma in adults.
Melanoma has one of the fastest rising incidence rates of any cancer. It accounts for a small percentage of skin cancer cases but is responsible for the majority of skin cancer deaths. Early detection and treatment is key to improving survival; however, anxiety around missing early cases needs to be balanced against appropriate levels of referral and excision of benign lesions. Used in conjunction with clinical or dermoscopic suspicion of malignancy, or both, reflectance confocal microscopy (RCM) may reduce unnecessary excisions without missing melanoma cases. To determine the diagnostic accuracy of reflectance confocal microscopy for the detection of cutaneous invasive melanoma and atypical intraepidermal melanocytic variants in adults with any lesion suspicious for melanoma and lesions that are difficult to diagnose, and to compare its accuracy with that of dermoscopy. We undertook a comprehensive search of the following databases from inception up to August 2016: Cochrane Central Register of Controlled Trials; MEDLINE; Embase; and seven other databases. We studied reference lists and published systematic review articles. Studies of any design that evaluated RCM alone, or RCM in comparison to dermoscopy, in adults with lesions suspicious for melanoma or atypical intraepidermal melanocytic variants, compared with a reference standard of either histological confirmation or clinical follow-up. Two review authors independently extracted all data using a standardised data extraction and quality assessment form (based on QUADAS-2). We contacted authors of included studies where information related to the target condition or diagnostic threshold were missing. We estimated summary sensitivities and specificities per algorithm and threshold using the bivariate hierarchical model. To compare RCM with dermoscopy, we grouped studies by population (defined by difficulty of lesion diagnosis) and combined data using hierarchical summary receiver operating characteristic (SROC) methods. Analysis of studies allowing direct comparison between tests was undertaken. To facilitate interpretation of results, we computed values of specificity at the point on the SROC curve with 90% sensitivity as this value lies within the estimates for the majority of analyses. We investigated the impact of using a purposely developed RCM algorithm and in-person test interpretation. The search identified 18 publications reporting on 19 study cohorts with 2838 lesions (including 658 with melanoma), which provided 67 datasets for RCM and seven for dermoscopy. Studies were generally at high or unclear risk of bias across almost all domains and of high or unclear concern regarding applicability of the evidence. Selective participant recruitment, lack of blinding of the reference test to the RCM result, and differential verification were particularly problematic. Studies may not be representative of populations eligible for RCM, and test interpretation was often undertaken remotely from the patient and blinded to clinical information.Meta-analysis found RCM to be more accurate than dermoscopy in studies of participants with any lesion suspicious for melanoma and in participants with lesions that were more difficult to diagnose (equivocal lesion populations). Assuming a fixed sensitivity of 90% for both tests, specificities were 82% for RCM and 42% for dermoscopy for any lesion suspicious for melanoma (9 RCM datasets; 1452 lesions and 370 melanomas). For a hypothetical population of 1000 lesions at the median observed melanoma prevalence of 30%, this equated to a reduction in unnecessary excisions with RCM of 280 compared to dermoscopy, with 30 melanomas missed by both tests. For studies in equivocal lesions, specificities of 86% would be observed for RCM and 49% for dermoscopy (7 RCM datasets; 1177 lesions and 180 melanomas). At the median observed melanoma prevalence of 20%, this reduced unnecessary excisions by 296 with RCM compared with dermoscopy, with 20 melanomas missed by both tests. Across all populations, algorithms and thresholds assessed, the sensitivity and specificity of the Pellacani RCM score at a threshold of three or greater were estimated at 92% (95% confidence interval (CI) 87 to 95) for RCM and 72% (95% CI 62 to 81) for dermoscopy. RCM may have a potential role in clinical practice, particularly for the assessment of lesions that are difficult to diagnose using visual inspection and dermoscopy alone, where the evidence suggests that RCM may be both more sensitive and specific in comparison to dermoscopy. Given the paucity of data to allow comparison with dermoscopy, the results presented require further confirmation in prospective studies comparing RCM with dermoscopy in a real-world setting in a representative population.


Human Capital Mediates Natural Selection in Contemporary Humans.
Natural selection has been documented in contemporary humans, but little is known about the mechanisms behind it. We test for natural selection through the association between 33 polygenic scores and fertility, across two generations, using data from UK Biobank (N = 409,629 British subjects with European ancestry). Consistently over time, polygenic scores that predict higher earnings, education and health also predict lower fertility. Selection effects are concentrated among lower SES groups, younger parents, people with more lifetime sexual partners, and people not living with a partner. The direction of natural selection is reversed among older parents, or after controlling for age at first live birth. These patterns are in line with the economic theory of fertility, in which earnings-increasing human capital may either increase or decrease fertility via income and substitution effects in the labour market. Studying natural selection can help us understand the genetic architecture of health outcomes: we find evidence in modern day Great Britain for multiple natural selection pressures that vary between subgroups in the direction and strength of their effects, that are strongly related to the socio-economic system, and that may contribute to health inequalities across income groups.


Large-margin classification in infinite neural networks.
We introduce a new family of positive-definite kernels for large margin classification in support vector machines (SVMs). These kernels mimic the computation in large neural networks with one layer of hidden units. We also show how to derive new kernels, by recursive composition, that may be viewed as mapping their inputs through a series of nonlinear feature spaces. These recursively derived kernels mimic the computation in deep networks with multiple hidden layers. We evaluate SVMs with these kernels on problems designed to illustrate the advantages of deep architectures. Compared to previous benchmarks, we find that on some problems, these SVMs yield state-of-the-art results, beating not only other SVMs but also deep belief nets.


Survey of parasites in domestic pigeons (Columba livia) in Tripoli, Libya.
This study was a preliminary survey for identification of parasites (endoparasites and ectoparasites) and their prevalence in domestic pigeons (Columba livia) in Tripoli; Libya. One hundred adult domestic pigeons were purchased from different markets of Tripoli. Blood smears were prepared from each bird and examined using light microscopy with oil immersion lens for the presence of blood parasites. Oropharyngeal swab was taken from each pigeon and were processed through direct smear method and staining with Giemsa to identify Trichomonas gallinae. Each pigeon was examined for ectoparasites. Moreover, intestinal parasites were investigated and all helminths were counted before being fixed in 70% ethyl alcohol for further identification. Out of 100 pigeons examined, 76% (76/100) were infected with Haemoproteus spp. and 55% (55/100) with T. gallinae. The overall prevalence of ecto-parasitic infestation was 89% (82% Columbicloa columbae, 18% Goniodes gallinae, 3% Menopon gallinae and 1% Pseudolynchia canariensis). The overall prevalence of intestinal helminths in examined pigeons were 56% (56/100). Three species of Cestoda (2% Raillietina tetragona, 32% R. echinobothrida and 4% R. cesticillus) and three species of Nematoda (18% Heterakis gallinarum., 22% Ascaridia galli and 4% Capillaria spp.) were identified. Also, this study revealed that the pigeons examined were found infected with mixed parasites. The study highlights that pigeons are hosts of helminths of veterinary importance and may act as an important source of infection for other avian host, which share the common parasitic fauna.


Importance of mosquito "quasispecies" in selecting an epidemic arthropod-borne virus.
Most arthropod-borne viruses (arboviruses), perpetuated by alternation between a vertebrate host and an insect vector, are likely to emerge through minor genetic changes enabling the virus to adapt to new hosts. In the past decade, chikungunya virus (CHIKV; Alphavirus, Togaviridae) has emerged on La Réunion Island following the selection of a unique substitution in the CHIKV E1 envelope glycoprotein (E1-A226V) of an East-Central-South African (ECSA) genotype conferring a higher transmission rate by the mosquito Aedes albopictus. Assumed to have occurred independently on at least four separate occasions, this evolutionary convergence was suspected to be responsible for CHIKV worldwide expansion. However, assumptions on CHIKV emergence were mainly based on viral genetic changes and the role of the mosquito population quasispecies remained unexplored. Here we show that the nature of the vector population is pivotal in selecting the epidemic CHIKV. We demonstrate using microsatellites mosquito genotyping that Ae. albopictus populations are genetically differentiated, contributing to explain their differential ability to select the E1-226V mutation. Aedes albopictus, newly introduced in Congo coinciding with the first CHIKV outbreak, was not able to select the substitution E1-A226V nor to preferentially transmit a CHIKV clone harboring the E1-226V as did Ae. albopictus from La Réunion.


Neonatal near miss approach in the 2005 WHO Global Survey Brazil.
To explore the use of the neonatal near miss concept as a tool to evaluate the quality of neonatal care, as 3 million early neonatal deaths occur every year around the world and the majority of these deaths are avoidable and take place in developing countries. This is a secondary analysis of the 2005 WHO Global Survey on Maternal and Perinatal Health, a cross-sectional study, using data from 19 randomly selected Brazilian hospitals. A pragmatic definition of neonatal near miss was developed and tested. Near miss indicators were calculated. Among the 15,169 live born infants included in this analysis, 424 presented at least one of the following conditions: very low birth weight, less than 30 gestational weeks at birth or an Apgar score at the 5th minute of life less than 7. According to the operational definition, these survivors from life-threatening conditions were considered neonatal near miss cases. The early neonatal mortality rate was 8.2/1,000 live births, the neonatal near miss rate was 21.4 neonatal near miss cases/1,000 live births. Substantial variations in the mortality among neonates with life-threatening conditions at birth were observed suggesting intra-hospital quality of care issues. The near miss concept and indicators provided information that could be useful to evaluate the quality of care and set priorities for further assessments and health care improvement for newborn infants.


Results of the ICTuS 2 Trial (Intravascular Cooling in the Treatment of Stroke 2).
Therapeutic hypothermia is a potent neuroprotectant approved for cerebral protection after neonatal hypoxia-ischemia and cardiac arrest. Therapeutic hypothermia for acute ischemic stroke is safe and feasible in pilot trials. We designed a study protocol to provide safer, faster therapeutic hypothermia in stroke patients. Safety procedures and 4°C saline infusions for faster cooling were added to the ICTuS trial (Intravascular Cooling in the Treatment of Stroke) protocol. A femoral venous intravascular cooling catheter after intravenous recombinant tissue-type plasminogen activator in eligible patients provided 24 hours cooling followed by a 12-hour rewarm. Serial safety assessments and imaging were performed. The primary end point was 3-month modified Rankin score 0,1. Of the intended 1600 subjects, 120 were enrolled before the study was stopped. Randomly, 63 were to receive hypothermia plus antishivering treatment and 57 normothermia. Compared with previous studies, cooling rates were improved with a cold saline bolus, without fluid overload. The intention-to-treat primary outcome of 90-day modified Rankin Score 0,1 occurred in 33% hypothermia and 38% normothermia subjects, odds ratio (95% confidence interval) of 0.81 (0.36-1.85). Serious adverse events occurred equally. Mortality was 15.9% hypothermia and 8.8% normothermia subjects, odds ratio (95% confidence interval) of 1.95 (0.56-7.79). Pneumonia occurred in 19% hypothermia versus 10.5% in normothermia subjects, odds ratio (95% confidence interval) of 1.99 (0.63-6.98). Intravascular therapeutic hypothermia was confirmed to be safe and feasible in recombinant tissue-type plasminogen activator-treated acute ischemic stroke patients. Protocol changes designed to reduce pneumonia risk appeared to fail, although the sample is small. URL: http://www.clinicaltrials.gov. Unique identifier: NCT01123161.


Sphingolipid-mediated inflammatory signaling leading to autophagy inhibition converts erythropoiesis to myelopoiesis in human hematopoietic stem/progenitor cells.
Elevated levels of the pro-inflammatory cytokine tumor necrosis factor-α (TNFα) inhibit erythropoiesis and cause anemia in patients with cancer and chronic inflammatory diseases. TNFα is also a potent activator of the sphingomyelinase (SMase)/ceramide pathway leading to ceramide synthesis and regulating cell differentiation, proliferation, apoptosis, senescence, and autophagy. Here we evaluated the implication of the TNFα/SMase/ceramide pathway on inhibition of erythropoiesis in human CD34+ hematopoietic stem/progenitor cells (CD34/HSPCs) from healthy donors. Exogenous synthetic C2- and C6-ceramide as well as bacterial SMase inhibited erythroid differentiation in erythropoietin-induced (Epo)CD34/HSPCs shown by the analysis of various erythroid markers. The neutral SMase inhibitor GW4869 as well as the genetic inhibition of nSMase with small interfering RNA (siRNA) against sphingomyelin phosphodiesterase 3 (SMPD3) prevented the inhibition by TNFα, but not the acid SMase inhibitor desipramine. Moreover, sphingosine-1-phosphate (S1P), a ceramide metabolite, restored erythroid differentiation, whereas TNFα inhibited sphingosine kinase-1, required for S1P synthesis. Analysis of cell morphology and colony formation demonstrated that erythropoiesis impairment was concomitant with a granulomonocytic differentiation in TNFα- and ceramide-treated EpoCD34/HSPCs. Inhibition of erythropoiesis and induction of granulomonocytic differentiation were correlated to modulation of hematopoietic transcription factors (TFs) GATA-1, GATA-2, and PU.1. Moreover, the expression of microRNAs (miR)-144/451, miR-146a, miR-155, and miR-223 was also modulated by TNFα and ceramide treatments, in line with cellular observations. Autophagy plays an essential role during erythropoiesis and our results demonstrate that the TNFα/neutral SMase/ceramide pathway inhibits autophagy in EpoCD34/HSPCs. TNFα- and ceramide-induced phosphorylation of mTORS2448 and ULK1S758, inhibited Atg13S355 phosphorylation, and blocked autophagosome formation as shown by transmission electron microscopy and GFP-LC3 punctae formation. Moreover, rapamycin prevented the inhibitory effect of TNFα and ceramides on erythropoiesis while inhibiting induction of myelopoiesis. In contrast, bafilomycin A1, but not siRNA against Atg5, induced myeloid differentiation, while both impaired erythropoiesis. We demonstrate here that the TNFα/neutral SMase/ceramide pathway inhibits erythropoiesis to induce myelopoiesis via modulation of a hematopoietic TF/miR network and inhibition of late steps of autophagy. Altogether, our results reveal an essential role of autophagy in erythroid vs. myeloid differentiation.


Five rules for the evolution of cooperation.
Cooperation is needed for evolution to construct new levels of organization. Genomes, cells, multicellular organisms, social insects, and human society are all based on cooperation. Cooperation means that selfish replicators forgo some of their reproductive potential to help one another. But natural selection implies competition and therefore opposes cooperation unless a specific mechanism is at work. Here I discuss five mechanisms for the evolution of cooperation: kin selection, direct reciprocity, indirect reciprocity, network reciprocity, and group selection. For each mechanism, a simple rule is derived that specifies whether natural selection can lead to cooperation.


Generation and genetic stability of tick-borne encephalitis virus mutants dependent on processing by the foot-and-mouth disease virus 3C protease.
Mature protein C of tick-borne encephalitis virus (TBEV) is cleaved from the polyprotein precursor by the viral NS2B/3 protease (NS2B/3(pro)). We showed previously that replacement of the NS2B/3(pro) cleavage site at the C terminus of protein C by the foot-and-mouth disease virus (FMDV) 2A StopGo sequence leads to the production of infectious virions. Here, we show that infectious virions can also be produced from a TBEV mutant bearing an inactivated 2A sequence through the expression of the FMDV 3C protease (3C(pro)) either in cis or in trans (from a TBEV replicon). Cleavage at the C terminus of protein C depended on the catalytic activity of 3C(pro) as well as on the presence of an optimized 3C(pro) cleavage site. Passage of the TBEV mutants bearing a 3C(pro) cleavage site either in the absence of 3C(pro) or in the presence of a catalytically inactive 3C(pro) led to the appearance of revertants in which protein C cleavage by NS2B/3(pro) had been regained. In three different revertants, a cleavage site for NS2B/3(pro), namely RR*C, was now present, leading to an elongated protein C. Furthermore, two revertants acquired additional mutations in the C terminus of protein C, eliminating two basic residues. Although these latter mutants showed wild-type levels of early RNA synthesis, their foci were smaller and an accumulation of protein C in the cytoplasm was observed. These findings suggest a role of the positive charge of the C terminus of protein C for budding of the nucleocapsid and further support the notion that TBEV protein C is a multifunctional protein.


Using speech sounds to test functional spectral resolution in listeners with cochlear implants.
In this study, spectral properties of speech sounds were used to test functional spectral resolution in people who use cochlear implants (CIs). Specifically, perception of the /ba/-/da/ contrast was tested using two spectral cues: Formant transitions (a fine-resolution cue) and spectral tilt (a coarse-resolution cue). Higher weighting of the formant cues was used as an index of better spectral cue perception. Participants included 19 CI listeners and 10 listeners with normal hearing (NH), for whom spectral resolution was explicitly controlled using a noise vocoder with variable carrier filter widths to simulate electrical current spread. Perceptual weighting of the two cues was modeled with mixed-effects logistic regression, and was found to systematically vary with spectral resolution. The use of formant cues was greatest for NH listeners for unprocessed speech, and declined in the two vocoded conditions. Compared to NH listeners, CI listeners relied less on formant transitions, and more on spectral tilt. Cue-weighting results showed moderately good correspondence with word recognition scores. The current approach to testing functional spectral resolution uses auditory cues that are known to be important for speech categorization, and can thus potentially serve as the basis upon which CI processing strategies and innovations are tested.


L-arginine augments endothelium-dependent vasodilation in cholesterol-fed rabbits.
Evidence exists that an endothelium-derived relaxing factor is nitric oxide and that L-arginine is the precursor for the synthesis of nitric oxide in vitro. Whether exogenous L-arginine contributes to the modulation of vascular smooth muscle tone in vivo is still controversial. In hypercholesterolemia, resistance vessels do not relax normally in response to pharmacological stimuli that release endothelium-derived relaxing factor; bioassay experiments have suggested that impaired synthesis or release of endothelium-derived relaxing factor accounts, in part, for this blunted relaxation. We hypothesized that hypercholesterolemia reduces arginine metabolism and thereby impairs endothelium-derived relaxing factor synthesis. Accordingly, we designed a study to determine whether exogenous L-arginine could augment endothelium-dependent vasodilation of hind limb resistance vessels in anesthetized cholesterol-fed rabbits. Femoral blood flow was recorded with an electromagnetic flow probe in 16 cholesterol-fed and 12 control rabbits. The hind limb vasodilator responses to incremental intra-arterial infusions of acetylcholine (0.3-9.0 micrograms/kg/min) and nitroprusside (0.3-9.0 micrograms/kg/min) were studied before and during intravenous administration of L-arginine (10 mg/kg/min), D-arginine (10 mg/kg/min), or saline. The vasodilator response to acetylcholine was impaired in cholesterol-fed rabbits as compared with control rabbits. L-Arginine augmented vasodilation to acetylcholine in cholesterol-fed but not in control rabbits. L-Arginine did not alter the effect of nitroprusside in either group. Neither saline nor D-arginine changed the response to either acetylcholine or nitroprusside. Our data demonstrate that exogenous L-arginine normalizes the endothelium-dependent vasodilation of hind limb resistance vessels in cholesterol-fed rabbits.


Optical sphere paint and a working standard of reflectance.
Specially prepared barium sulfate powders and coatings of these powders with polyvinyl alcohol have been used as working standards of reflectance and for coating integrating spheres. These materials are satisfactory for use in the wavelength range 0.20-2.00 micro. Since the refractive index of barium sulfate is rather low, it is necessary to use fairly thick layers in order to obtain good results. The absolute value of luminous reflectance of the barium sulfate powder, described in this paper, is 0.995 +/- 0.001, and that of the paint, when properly applied, is 0.992 +/- 0.001. The spectral reflectance of this material is higher than that of magnesium oxide, particularly in the uv region of the spectrum. By using barium sulfate as reference standard and as a coating on the integrating spheres of spectrophotometers, it is now possible to measure spectral reflectance reliably in the uv region to 200 nm. Because the reflectance of barium sulfate paint is significantly greater than that of magnes um oxide in the shortwave region, there is a corresponding gain in instrument sensitivity when barium sulfate is used as a sphere coating and as a comparison material.


Analysis of copy number changes suggests chromosomal instability in a minority of large colorectal adenomas.
We have examined chromosomal-scale mutations in 34 large colorectal adenomas (CRAs). A small number of changes (median = 2, IQR = 0-4) were found by array-comparative genomic hybridization (aCGH) in most tumours. The most common changes were deletions of chromosomes 1p, 9q, 17, 19, and 22, and gains of chromosomes 13 and 21. SNP-LOH analysis and pseudo-digital SNP-PCR analysis detected occasional copy-neutral LOH. Some aCGH changes found frequently in colorectal carcinomas, such as deletions of chromosomes 4q and 18q, were very infrequent in the adenomas. Almost all copy number changes were of small magnitude, far below the predicted levels even for single copy gain/loss; investigation suggested that these changes were either artefactual or occurred in sub-clones within the tumours. In some cases, these sub-clones may have represented progression towards carcinoma, but comparison with aCGH data from carcinomas showed this to be unlikely in most cases. In two adenomas, there was evidence of a large, outlying number of copy number changes, mostly resulting from part-chromosome deletions. Overall, moreover, there was evidence of a tendency towards part-chromosome deletions-consistent with chromosomal instability (CIN)--in about one-sixth of all tumours. However, there was no evidence of CIN in the form of whole-chromosome copy number changes. Our data did not support previous contentions that CRAs tend to show chromosome breakage at fragile sites owing to CIN associated with an elevated DNA damage response. Chromosomal-scale mutations occur in some CRAs; although CIN is not the norm in these lesions, it probably affects a minority of cases.


Impact of adjusting for inter-rater variability in conference abstract ranking and selection processes.
Scientific conferences provide a forum for clinicians, educators, students and researchers to share research findings. To be selected to present at a scientific conference, authors must submit a short abstract which is then rated on its scientific quality and professional merit and is accepted or rejected based on these ratings. Previous research has indicated that inter-rater variability can have a substantial impact on abstract selection decisions. For their 2015 conference, the Occupational Therapy Australia National Conference introduced a system to identify and adjust for inter-rater variability in the abstract ranking and selection process. Ratings for 1340 abstracts submitted for the 2015 and 2017 conferences were analysed using many-faceted Rasch analysis to identify and adjust for inter-rater variability. Analyses of the construct validity of the abstract rating instrument and rater consistency were completed. To quantify the influence of inter-rater variability of abstract selection decisions, comparisons were made between decisions made using Rasch-calibrated measure scores and decisions that would have been made based purely on raw average scores derived from the abstract ratings. Construct validity and measurement properties of the abstract rating tool were good to excellent (item fit MnSq scores ranged from 0.8 to 1.2; item reliability index = 1.0). Most raters (24 of 27, 89%) were consistent in their use of the rating instrument. When comparing abstract allocations under the two conditions, 25% of abstracts (n = 341) would have been allocated differently if inter-rater variability was not accounted for. This study demonstrates that, even with a strong abstract rating instrument and a small rater pool, inter-rater variability still exerts a substantial influence on abstract selection decisions. It is recommended that all occupational therapy conferences internationally, and scientific conferences more generally, adopt systems to identify and adjust for the impact of inter-rater variability in abstract selection processes.


Transient inhibition of TrkB kinase after status epilepticus prevents development of temporal lobe epilepsy.
Temporal lobe epilepsy is the most common and often devastating form of human epilepsy. The molecular mechanism underlying the development of temporal lobe epilepsy remains largely unknown. Emerging evidence suggests that activation of the BDNF receptor TrkB promotes epileptogenesis caused by status epilepticus. We investigated a mouse model in which a brief episode of status epilepticus results in chronic recurrent seizures, anxiety-like behavior, and destruction of hippocampal neurons. We used a chemical-genetic approach to selectively inhibit activation of TrkB. We demonstrate that inhibition of TrkB commencing after status epilepticus and continued for 2 weeks prevents recurrent seizures, ameliorates anxiety-like behavior, and limits loss of hippocampal neurons when tested weeks to months later. That transient inhibition commencing after status epilepticus can prevent these long-lasting devastating consequences establishes TrkB signaling as an attractive target for developing preventive treatments of epilepsy in humans.


Enhancement of T-cell-mediated antitumor response: angiostatic adjuvant to immunotherapy against cancer.
Tumor-released proangiogenic factors suppress endothelial adhesion molecule (EAM) expression and prevent leukocyte extravasation into the tumor. This is one reason why immunotherapy has met with limited success in the clinic. We hypothesized that overcoming EAM suppression with angiogenesis inhibitors would increase leukocyte extravasation and subsequently enhance the effectiveness of cellular immunotherapy. Intravital microscopy, multiple color flow cytometry, immunohistochemistry, and various tumor mouse (normal and T-cell deficient) models were used to investigate the temporal dynamics of cellular and molecular events that occur in the tumor microenvironment during tumor progression and angiostatic intervention. We report that while EAM levels and T-cell infiltration are highly attenuated early on in tumor growth, angiostatic therapy modulates these effects. In tumor models with normal and T-cell-deficient mice, we show the active involvement of the adaptive immune system in cancer and differentiate antiangiogenic effects from antiangiogenic mediated enhancement of immunoextravasation. Our results indicate that a compromised immune response in tumors can be obviated by the use of antiangiogenic agents. Finally, with adoptive transfer studies in mice, we show that a phased combination of angiostatic therapy and T-cell transfer significantly (P < 0.0013) improves tumor growth inhibition. This research contributes to understand the cellular mechanism of action of angiostatic agents and the immune response within the tumor microenvironment, in particular as a consequence of the temporal dynamics of EAM levels. Moreover, our results suggest that adjuvant therapy with angiogenesis inhibitors holds promise for cellular immunotherapy in the clinic.


Human cytomegalovirus hijacks host stress response fueling replication stress and genome instability.
Viral infections enhance cancer risk and threaten host genome integrity. Although human cytomegalovirus (HCMV) proteins have been detected in a wide spectrum of human malignancies and HCMV infections have been implicated in tumorigenesis, the underlying mechanisms remain poorly understood. Here, we employed a range of experimental approaches, including single-molecule DNA fiber analysis, and showed that infection by any of the four commonly used HCMV strains: AD169, Towne, TB40E or VR1814 induced replication stress (RS), as documented by host-cell replication fork asymmetry and formation of 53BP1 foci. The HCMV-evoked RS triggered an ensuing host DNA damage response (DDR) and chromosomal instability in both permissive and non-permissive human cells, the latter being particularly relevant in the context of tumorigenesis, as such cells can survive and proliferate after HCMV infection. The viral major immediate early enhancer and promoter (MIEP) that controls expression of the viral genes IE72 (IE-1) and IE86 (IE-2), contains transcription-factor binding sites shared by promoters of cellular stress-response genes. We found that DNA damaging insults, including those relevant for cancer therapy, enhanced IE72/86 expression. Thus, MIEP has been evolutionary shaped to exploit host DDR. Ectopically expressed IE72 and IE86 also induced RS and increased genomic instability. Of clinical relevance, we show that undergoing standard-of-care genotoxic radio-chemotherapy in patients with HCMV-positive glioblastomas correlated with elevated HCMV protein markers after tumor recurrence. Collectively, these results are consistent with our proposed concept of HCMV hijacking transcription-factor binding sites shared with host stress-response genes. We present a model to explain the potential oncomodulatory effects of HCMV infections through enhanced replication stress, subverted DNA damage response and induced genomic instability.


Overexpression of PIK3CA in murine head and neck epithelium drives tumor invasion and metastasis through PDK1 and enhanced TGFβ signaling.
Head and neck squamous cell carcinoma (HNSCC) patients have a poor prognosis, with invasion and metastasis as major causes of mortality. The phosphatidylinositol 3-kinase (PI3K) pathway regulates a wide range of cellular processes crucial for tumorigenesis, and PIK3CA amplification and mutation are among the most common genetic alterations in human HNSCC. Compared with the well-documented roles of the PI3K pathway in cell growth and survival, the roles of the PI3K pathway in tumor invasion and metastasis have not been well delineated. We generated a PIK3CA genetically engineered mouse model (PIK3CA-GEMM) in which wild-type PIK3CA is overexpressed in head and neck epithelium. Although PIK3CA overexpression alone was not sufficient to initiate HNSCC formation, it significantly increased tumor susceptibility in an oral carcinogenesis mouse model. PIK3CA overexpression in mouse oral epithelium increased tumor invasiveness and metastasis by increasing epithelial-mesenchymal transition and by enriching a cancer stem cell phenotype in tumor epithelial cells. In addition to these epithelial alterations, we also observed marked inflammation in tumor stroma. AKT is a central signaling mediator of the PI3K pathway. However, molecular analysis suggested that progression of PIK3CA-driven HNSCC is facilitated by 3-phosphoinositide-dependent protein kinase (PDK1) and enhanced transforming growth factor β (TGFβ) signaling rather than by AKT. Examination of human HNSCC clinical samples revealed that both PIK3CA and PDK1 protein levels correlated with tumor progression, highlighting the significance of this pathway. In summary, our results offer significant insight into how PIK3CA overexpression drives HNSCC invasion and metastasis, providing a rationale for targeting PI3K/PDK1 and TGFβ signaling in advanced HNSCC patients with PIK3CA amplification.


Setting healthcare priorities: a description and evaluation of the budgeting and planning process in county hospitals in Kenya.
This paper describes and evaluates the budgeting and planning processes in public hospitals in Kenya. We used a qualitative case study approach to examine these processes in two hospitals in Kenya. We collected data by in-depth interviews of national level policy makers, hospital managers, and frontline practitioners in the case study hospitals (n = 72), a review of documents, and non-participant observations within the hospitals over a 7 month period. We applied an evaluative framework that considers both consequentialist and proceduralist conditions as important to the quality of priority-setting processes. The budgeting and planning process in the case study hospitals was characterized by lack of alignment, inadequate role clarity and the use of informal priority-setting criteria. With regard to consequentialist conditions, the hospitals incorporated economic criteria by considering the affordability of alternatives, but rarely considered the equity of allocative decisions. In the first hospital, stakeholders were aware of - and somewhat satisfied with - the budgeting and planning process, while in the second hospital they were not. Decision making in both hospitals did not result in reallocation of resources. With regard to proceduralist conditions, the budgeting and planning process in the first hospital was more inclusive and transparent, with the stakeholders more empowered compared to the second hospital. In both hospitals, decisions were not based on evidence, implementation of decisions was poor and the community was not included. There were no mechanisms for appeals or to ensure that the proceduralist conditions were met in both hospitals. Public hospitals in Kenya could improve their budgeting and planning processes by harmonizing these processes, improving role clarity, using explicit priority-setting criteria, and by incorporating both consequentialist (efficiency, equity, stakeholder satisfaction and understanding, shifted priorities, implementation of decisions), and proceduralist (stakeholder engagement and empowerment, transparency, use of evidence, revisions, enforcement, and incorporating community values) conditions.


Intra-articular hyaluronate, tenoxicam and vitamin E in a rat model of osteoarthritis: evaluation and comparison of chondroprotective efficacy.
The aim of this experimental study was to evaluate and compare the chondroprotective efficacy of intra-articular hyaluronic acid, tenoxicam and vitamin E in osteoarthritis. An osteoarthritis model was created by anterior cruciate ligament transection and medial menisectomy in knees of 28 rats. The rats were randomized into four groups; first group served as a control group and received intra-articular injections of saline solution, intra-articular HA, intra-articular tenoxicam and intra-articular Vit E were applied to the treatment groups. First intra-articular injections were applied at second week postoperatively and repeated once a week for 5 weeks. At 8th week after the operation groups were compared based on the histologic scores of cartilage degeneration by Mankin Histological Grading Scale. Total cartilage degeneration score was significantly increased in the control group (P=0.004). Total Mankin scores of HA, tenoxicam and Vit E groups were significantly lower than the control group (P=0.004, P=0.016, P=0.012 respectively). There was no statistically siginificant difference between the treatment groups in terms of total Mankin scores (P>0.05). Intra-articular application of HA, tenoxicam and Vit E are chondroprotective in early osteoarthritis model in rats. Chondroprotective activity of tenoxicam and Vit E are comparable with the beneficial effects of HA on articular cartilage.


Reducing Cervical Retrolisthesis With Long-Term Monthly Chiropractic Maintenance Care: A Case Report.
The present case study aimed to demonstrate the subtle effectiveness of 13-year chiropractic maintenance care (MC) in a patient with cervical radiculopathy associated with retrolisthesis. Retrolisthesis is the backward slippage of one vertebral body with respect to the subjacent vertebra. This slippage can lead to spinal nerve root compression or irritation (radiculopathy). A 44-year-old woman presented with relapsing episodes of pain in her neck radiating down her right arm and hand for 12 months. Neurological examination revealed sensory deficits in the right C5 dermatome. Cervical radiography showed grade 1 retrolisthesis of C3 on C4 and of C4 on C5, and sclerosis over the upper endplate of the C4, C5, and C6 vertebrae. The patient was diagnosed with right C5 radiculopathy associated with vertebral retrolisthesis. She was conservatively treated with multimodal chiropractic care comprising cervical manipulation applied to the affected segments, axial distraction, and isometric stretching. After 20 sessions over 3 months of therapy, the patient experienced complete relief of neck pain and radicular symptoms, and full cervical mobility. Because her complaints were characterized by relapsing episodes, she was enrolled in a monthly MC program to monitor and ensure the spine was functioning at its highest capacity. The MC program included disease monitoring, manipulation of dysfunctional segments, cervical strengthening exercises, and ergonomics coaching. Monthly care was shown to be successful in maintaining an asymptomatic status. Moreover, a gradual reduction in cervical retrolisthesis was observed during the 13-year MC period, and the ongoing MC ensured optimal spine functioning.


A degenerate HLA-DR epitope pool of HER-2/neu reveals a novel in vivo immunodominant epitope, HER-2/neu88-102.
Over the past two decades, there has been significant interest in targeting HER-2/neu in immune-based approaches for the treatment of HER-2/neu+ cancers. For example, peptide vaccination using a CD8 T cell-activating HER-2/neu epitope (amino acids 369-377) is an approach that is being considered in advanced phase clinical trials. Studies have suggested that the persistence of HER-2/neu-specific CD8 T cells could be improved by incorporating human leukocyte antigen (HLA) class II epitopes in the vaccine. Our goal in this study was to identify broad coverage HLA-DR epitopes of HER-2/neu, an antigen that is highly expressed in a variety of carcinomas. A combination of algorithms and HLA-DR-binding assays was used to identify HLA-DR epitopes of HER-2/neu antigen. Evidence of preexistent immunity in cancer patients against the identified epitopes was determined using IFN-gamma enzyme-linked immunosorbent spot (ELIspot) assay. Eighty-four HLA-DR epitopes of HER-2/neu were predicted, 15 of which had high binding affinity for > or =11 common HLA-DR molecules. A degenerate pool of four HLA-DR-restricted 15-amino acid epitopes (p59, p88, p422, and p885) was identified, against which >58% of breast and ovarian cancer patients had preexistent T-cell immunity. All four epitopes are naturally processed by antigen-presenting cells. Hardy-Weinberg analysis showed that the pool is useful in approximately 84% of population. Lastly, in this degenerate pool, we identified a novel in vivo immunodominant HLA-DR epitope, HER-2/neu(88-102) (p88). The broad coverage and natural immunity to this epitope pool suggests potential usefulness in HER-2/neu-targeting, immune-based therapies such as vaccines.


The Akt-nitric oxide-cGMP pathway contributes to nerve growth factor-mediated neurite outgrowth in apolipoprotein E knockout mice.
Apolipoprotein E (apo)-deficient [apoE(-/-)] mice have peripheral sensory nerve defects and a reduced and delayed response to noxious thermal stimuli. However, to date, no report has focused on the influence of apoE deficiency on calcitonin gene-related peptide (CGRP)-containing nerve fiber extensions. We have shown that the density of CGRP-containing nerve fibers decreases in mesenteric arteries of apoE(-/-) mice compared with wild-type mice. Here, we investigated whether apoE deficiency is involved in nerve growth factor (NGF)-induced CGRP-containing nerve regeneration using apoE(-/-) mice. NGF-mediated CGRP-like immunoreactivity (LI)-neurite outgrowth in apoE(-/-) cultured dorsal root ganglia (DRG) cells was significantly lower than that in wild-type cultures. However, the level of NGF receptor mRNA in apoE(-/-) DRG cells was similar to that in wild-type mice. To clarify the mechanism of the impaired ability of NGF-mediated neurite outgrowth, we focused on the Akt-nitric oxide (NO)-cGMP pathway. Expression of phosphorylated Akt was significantly reduced in apoE(-/-) DRG. The NO donor, sodium nitroprusside or S-nitroso-N-acetylpenicillamine, did not affect NGF-mediated neurite outgrowth in apoE(-/-) cultured DRG cells. However, 8-bromoguanosine 3',5'-cyclic monophosphate sodium salt n-hydrate, a cGMP analog, induced NGF-mediated nerve facilitation similar to wild-type NGF-mediated neurite outgrowth levels. Furthermore, in apoE(-/-) DRG, soluble guanylate cyclase expression was significantly lower than that in wild-type DRG. These results suggest that in apoE(-/-) mice the Akt-NO-cGMP pathway is impaired, which may be caused by NGF-mediated CGRP-LI-neurite outgrowth defects.


An empirical comparison of the OPQoL-Brief, EQ-5D-3 L and ASCOT in a community dwelling population of older people.
This study examined the relationships between a newly developed older person-specific non-preference-based quality of life (QoL) instrument (Older People's Quality of Life brief questionnaire (OPQoL-brief)) and two generic preference-based instruments (the EQ-5D-3L Level (EQ-5D-3 L) and the Adult Social Care Outcomes Toolkit (ASCOT) in a community-dwelling population of Australian older people receiving aged care services. We formulated hypotheses about the convergent validity between the instruments (examined by Wilcoxon-Mann Whitney, Kruskal Wallis and Spearman's correlation tests) and levels of agreement (assessed using intra class correlation (ICC) and modified Bland-Altman plots based on normalized Z EQ-5D-3 L and ASCOT utilities and OPQoL-Brief summary scores). The utilities/summary scores for 87 participants (aged 65-93 years) were moderately but positively correlated. Moderate convergent validity was evident for a number of instrument dimensions with the strongest relationship (r = 0.57) between 'enjoy life' (OPQoL-Brief) and 'social contact' (ASCOT). The overall ICC was 0.54 and Bland-Altman scatter plots showed 3-6% of normalized Z-scores were outside the 95% limits of agreement suggesting moderate agreement between all three instruments (agreement highest between the OPQoL-Brief and the ASCOT). Our results suggest that the OPQoL-Brief, the ASCOT and the EQ-5D_3L are suitable for measuring quality of life outcomes in community-dwelling populations of older people. Given the different constructs underpinning these instruments, we recommend that choice of instrument should be guided by the context in which the instruments are being applied. Currently, the OPQoL-Brief is not suitable for use in cost-utility analyses as it is not preference-based. Given their different perspectives, we recommend that both the ASCOT and the EQ-5D are applied simultaneously to capture broader aspects of quality of life and health status within cost-utility analyses within the aged care sector. Future research directed towards the development of a new single preference-based instrument that incorporates both health status and broader aspects of quality of life within quality adjusted life year calculations for older people would be beneficial.


Optimal loading for peak power output during the hang power clean in professional rugby players.
The ability to develop high levels of muscle power is considered an essential component of success in many sporting activities; however, the optimal load for the development of peak power during training remains controversial. The aim of the present study was to determine the optimal load required to observe peak power output (PPO) during the hang power clean in professional rugby players. Twelve professional rugby players performed hang power cleans on a portable force platform at loads of 30%, 40%, 50%, 60%, 70%, 80%, and 90% of their predetermined 1-repetition maximum (1-RM) in a randomized and balanced order. Relative load had a significant effect on power output, with peak values being obtained at 80% of the subjects' 1-RM (4466 +/- 477 W; P < .001). There was no significant difference, however, between the power outputs at 50%, 60%, 70%, or 90% 1-RM compared with 80% 1-RM. Peak force was produced at 90% 1-RM with relative load having a significant effect on this variable; however, relative load had no effect on peak rate of force development or velocity during the hang power clean. The authors conclude that relative load has a significant effect on PPO during the hang power clean: Although PPO was obtained at 80% 1-RM, there was no significant difference between the loads ranging from 40% to 90% 1-RM. Individual determination of the optimal load for PPO is necessary in order to enhance individual training effects.


[Problematic sickness absence: a quantitative and qualitative analysis].
In the Netherlands, an occupational physician supports absence management and staff's return to work, but this role may be hampered if there is uncertainty concerning return to labour options. The return to work process may be improved when a specialized second opinion organization is called upon. Our objective was to map the potential for deployment, return to work rates, and healthcare costs for staff with problematic sickness absence profiles. We analysed the data of problematic absentees who passed through a second opinion advisory system between 2011-2014, using pseudo-anonymized data matching from various sources. The datasets include client data from a company providing second opinions (n = 2595), a questionnaire for employees (n = 1800), a questionnaire for referrers (n = 400), additional data from one case management company (n = 564) and claims data from a health insurance company (n = 725). Among the problematic absentees, 49% concerned musculoskeletal problems, compared to 23% mental health problems, 14% multifactorial problems, and 16% other problems; 29% concerned limitations restricting return to work. In 51% of the cases, physicians representing the second opinion company concluded that current treatment was inadequate. Fifty-three percent of the referrers responded that the second opinion usually led to a change in treatment. In 31% of cases, employees felt that the second opinion system did not contribute to the return to work process. Average annual health costs of problematic absentees were €7800, compared to €2600 before sickness absence commenced. Many employees with problematic sickness absence are not treated adequately; opportunities to return to work are not utilized and the treatment is often not sufficiently focused on functional recovery. In principle, (partial) return to work is deemed possible for the majority of cases.


Generic versus brand-name drugs used in cardiovascular diseases.
This meta-analysis aimed to compare the efficacy and adverse events, either serious or mild/moderate, of all generic versus brand-name cardiovascular medicines. We searched randomized trials in MEDLINE, Scopus, EMBASE, Cochrane Controlled Clinical Trial Register, and ClinicalTrials.gov (last update December 1, 2014). Attempts were made to contact the investigators of all potentially eligible trials. Two investigators independently extracted and analyzed soft (including systolic blood pressure, LDL cholesterol, and others) and hard efficacy outcomes (including major cardiovascular adverse events and death), minor/moderate and serious adverse events. We included 74 randomized trials; 53 reported ≥1 efficacy outcome (overall sample 3051), 32 measured mild/moderate adverse events (n = 2407), and 51 evaluated serious adverse events (n = 2892). We included trials assessing ACE inhibitors (n = 12), anticoagulants (n = 5), antiplatelet agents (n = 17), beta-blockers (n = 11), calcium channel blockers (n = 7); diuretics (n = 13); statins (n = 6); and others (n = 3). For both soft and hard efficacy outcomes, 100 % of the trials showed non-significant differences between generic and brand-name drugs. The aggregate effect size was 0.01 (95 % CI -0.05; 0.08) for soft outcomes; -0.06 (-0.71; 0.59) for hard outcomes. All but two trials showed non-significant differences in mild/moderate adverse events, and aggregate effect size was 0.07 (-0.06; 0.20). Comparable results were observed for each drug class and in each stratified meta-analysis. Overall, 8 serious possibly drug-related adverse events were reported: 5/2074 subjects on generics; 3/2076 subjects on brand-name drugs (OR 1.69; 95 % CI 0.40-7.20). This meta-analysis strengthens the evidence for clinical equivalence between brand-name and generic cardiovascular drugs. Physicians could be reassured about prescribing generic cardiovascular drugs, and health care organization about endorsing their wider use.


Precompensation of 3D field distortions in remote focus two-photon microscopy.
Remote focusing is widely used in 3D two-photon microscopy and 3D photostimulation because it enables fast axial scanning without moving the objective lens or specimen. However, due to the design constraints of microscope optics, remote focus units are often located in non-telecentric positions in the optical path, leading to significant depth-dependent 3D field distortions in the imaging volume. To address this limitation, we characterized 3D field distortions arising from non-telecentric remote focusing and present a method for distortion precompensation. We demonstrate its applicability for a 3D two-photon microscope that uses an acousto-optic lens (AOL) for remote focusing and scanning. We show that the distortion precompensation method improves the pointing precision of the AOL microscope to < 0.5 µm throughout the 400 × 400 × 400 µm imaging volume.


[Detection of antibacterial resistance by MALDI-TOF mass spectrometry].
In the last decade we have witnessed a remarkable increase in the number of strains isolated in hospitals that are producing extended spectrum β-lactamases (ESBL) or, more recently, carbapenemases. This makes clear the need for a system for rapid detection of these resistance mechanisms that allow the selection of the most suitable antibiotic treatment in order to improve patient care. Recent data support the possibility of using mass spectrometry (MS), specifically MALDI -TOF (Matrix-Assisted Laser Desorption / Ionization, Time-of-Flight ) systems to identify specific resistance mechanisms and their use offers several advantages. First, the economic cost of each determination is clearly inferior to the classical molecular techniques for detection of resistance genes. Second, detection of resistance by MALDI -TOF reduces the time for obtaining results compared to the routine methods currently employed. Finally, the possibility that this method allows us to detect enzymes not previously characterized, that there is no information about the genes that encode them. Therefore, we believe that this may be a good tool to implement in clinical microbiology laboratories. This review aims to present the latest developments in this field.


Home oxygen therapy reduces risk of hospitalisation in patients with chronic obstructive pulmonary disease: a population-based retrospective cohort study, 2005-2012.
This study evaluated the effect of home oxygen therapy (HOT) on hospital admissions in chronic obstructive pulmonary disease (COPD) patients. Using nationwide health insurance claims from 2002-2012, we conducted a longitudinal population-based retrospective cohort study. Individuals who were aged 40 years or above and newly diagnosed with COPD in 2005. The primary outcome was total number of hospitalisations during the study period. Participants were matched using HOT propensity scores and were stratified by respiratory impairment (grade 1: FEV1 ≤25% or PaO2 ≤55 mm Hg; grade 2: FEV1 ≤30% or PaO2 56-60 mm Hg; grade 3: FEV1 ≤40% or PaO2 61-65 mm Hg; 'no grade': FEV1 or PaO2 unknown), then a negative binomial regression analysis was performed for each group. Of the 36,761 COPD patients included in our study, 1330 (3.6%) received HOT. In a multivariate analysis of grade 1 patients performed before propensity score matching, the adjusted relative risk of hospitalisation for patients who did not receive HOT was 1.27 (95% CI 1.01 to 1.60). In a multivariate analysis of grade 1 patients performed after matching, the adjusted relative risk for patients who did not receive HOT was 1.65 (95% CI 1.25 to 2.18). In grade 2 or grade 3 patients, no statistical difference in hospital admission risk was detected. In the 'no grade' group of patients, HOT was associated with an increased risk of hospitalisation. HOT reduces the risk of hospital admission in COPD patients with severe hypoxaemia. However, apart from these patients, HOT use is not associated with hospital admissions.


Tritium uptake in rainbow trout (Oncorhynchus mykiss): HTO and OBT-spiked feed exposures simultaneously.
There is currently considerable interest in organically bound tritium (OBT) formation in edible fish. The major questions revolve around whether or not tritium can accumulate in fish after being released into aquatic environments. Since OBT formation rates in large, edible fish are poorly understood, rainbow trout (Oncorhynchus mykiss) studies, where fish were simultaneously exposed to tritiated water (HTO) and OBT-spiked feed over 130 days, were conducted to evaluate tritium uptake. The measured HTO activity concentrations in fish tissue confirmed that HTO in fish tissue equilibrates quickly with HTO in tank water. The data obtained also confirmed that OBT uptake is faster when fish are ingesting OBT-spiked feed compared to when fish are living in tritiated water (and consuming non-OBT-spiked feed). The difference between the two exposure types is such that the groups exposed to tritiated water and OBT-spiked feed simultaneously were showing the same uptake rates as OBT-spiked feed only exposures. Contrary to what was expected, the rate of OBT uptake (from OBT-spiked feed) seemed to be higher in slow growing fish compared to fast growing fish. Another observation from these studies was that OBT activity concentrations in all organs (viscera) had a tendency to be higher than OBT activity concentrations measured in fish flesh.


Effectiveness of a 'Do not interrupt' bundled intervention to reduce interruptions during medication administration: a cluster randomised controlled feasibility study.
To evaluate the effectiveness of a 'Do not interrupt' bundled intervention to reduce non-medication-related interruptions to nurses during medication administration. A parallel eight cluster randomised controlled study was conducted in a major teaching hospital in Adelaide, Australia. Four wards were randomised to the intervention which comprised wearing a vest when administering medications; strategies for diverting interruptions; clinician and patient education; and reminders. Control wards were blinded to the intervention. Structured direct observations of medication administration processes were conducted. The primary outcome was non-medication-related interruptions during individual medication dose administrations. The secondary outcomes were total interruption and multitasking rates. A survey of nurses' experiences was administered. Over 8 weeks and 364.7 hours, 227 nurses were observed administering 4781 medications. At baseline, nurses experienced 57 interruptions/100 administrations, 87.9% were unrelated to the medication task being observed. Intervention wards experienced a significant reduction in non-medication-related interruptions from 50/100 administrations (95% CI 45 to 55) to 34/100 (95% CI 30 to 38). Controlling for clustering, ward type and medication route showed a significant reduction of 15 non-medication-related interruptions/100 administrations compared with control wards. A total of 88 nurses (38.8%) completed the poststudy survey. Intervention ward nurses reported that vests were time consuming, cumbersome and hot. Only 48% indicated that they would support the intervention becoming hospital policy. Nurses experienced a high rate of interruptions. Few were related to the medication task, demonstrating considerable scope to reduce unnecessary interruptions. While the intervention was associated with a statistically significant decline in non-medication-related interruptions, the magnitude of this reduction and its likely impact on error rates should be considered, relative to the effectiveness of alternate interventions, associated costs, likely acceptability and long-term sustainability of such interventions.


Multiple system atrophy-associated oligodendroglial protein p25α stimulates formation of novel α-synuclein strain with enhanced neurodegenerative potential.
Pathology consisting of intracellular aggregates of alpha-Synuclein (α-Syn) spread through the nervous system in a variety of neurodegenerative disorders including Parkinson's disease, dementia with Lewy bodies, and multiple system atrophy. The discovery of structurally distinct α-Syn polymorphs, so-called strains, supports a hypothesis where strain-specific structures are templated into aggregates formed by native α-Syn. These distinct strains are hypothesised to dictate the spreading of pathology in the tissue and the cellular impact of the aggregates, thereby contributing to the variety of clinical phenotypes. Here, we present evidence of a novel α-Syn strain induced by the multiple system atrophy-associated oligodendroglial protein p25α. Using an array of biophysical, biochemical, cellular, and in vivo analyses, we demonstrate that compared to α-Syn alone, a substoichiometric concentration of p25α redirects α-Syn aggregation into a unique α-Syn/p25α strain with a different structure and enhanced in vivo prodegenerative properties. The α-Syn/p25α strain induced larger inclusions in human dopaminergic neurons. In vivo, intramuscular injection of preformed fibrils (PFF) of the α-Syn/p25α strain compared to α-Syn PFF resulted in a shortened life span and a distinct anatomical distribution of inclusion pathology in the brain of a human A53T transgenic (line M83) mouse. Investigation of α-Syn aggregates in brain stem extracts of end-stage mice demonstrated that the more aggressive phenotype of the α-Syn/p25α strain was associated with an increased load of α-Syn aggregates based on a Förster resonance energy transfer immunoassay and a reduced α-Syn aggregate seeding activity based on a protein misfolding cyclic amplification assay. When injected unilaterally into the striata of wild-type mice, the α-Syn/p25α strain resulted in a more-pronounced motoric phenotype than α-Syn PFF and exhibited a "tropism" for nigro-striatal neurons compared to α-Syn PFF. Overall, our data support a hypothesis whereby oligodendroglial p25α is responsible for generating a highly prodegenerative α-Syn strain in multiple system atrophy.


What is a comprehensive mental health nursing assessment? A review of the literature.
Assessment is the foundation on which nursing care is delivered. The aim of this paper is to better understand the content (what information nurses seek about consumers) and the process (how they go about gathering that information) of a comprehensive mental health nursing assessment in practice. Using terms, such as 'nursing', 'mental health', and 'assessment', the CINAHL, Medline, and PsycINFO databases were searched for studies that describe the content and process of a comprehensive mental health nursing assessment. Although studies of aspects of mental health nursing assessment, such as the assessment of risk or carer burden, were found, no single study described both the content and process of a comprehensive mental health nursing assessment in practice. In Australia, a comprehensive assessment is codified as a competency to practice nursing; however, the standards of practice set for mental health nurses are less clear on what constitutes a comprehensive assessment or how this should be undertaken. The peer-reviewed literature describes assessment as both an independent and interdependent activity. It is described as informal, and there is evidence that comprehensive mental health nursing assessments are not well documented. The credibility of training and research into mental health nursing requires that the content and the process of a comprehensive mental health nursing assessment needs to be clearly defined.


Vessel morphometric parameters-correlation with histologic grade and VEGF expression in oligodendroglioma.
The contributions of histologic features including microvascular proliferation to the determination of malignancy in oligodendrogliomas remain uncertain. We have retrospectively performed morphometric assessments in 20 tumors histologically classified as well-differentiated (WHO Grade II, n=8) or anaplastic (WHO Grade III, n=12) oligodendrogliomas (WDO or AO). Quantitative studies utilized image analysis of double immunolabeled vasculature with anti CD34 with VIP chromogen (purple) and proliferating nuclei with anti MIB-1, using DAB (brown). Mean values are reported from five fields for each of twenty cases. The total number of MIB-1 positive tumor nuclei was 10 fold higher in AO vs WDO. The area occupied by vessels was also markedly increased in AO vs WDO, as was the microvessel density. Proliferating endothelial cells i.e. those with MIB-1 positive nuclei in CD34 positive cells were significantly increased (4.6 vs 0.26 positive nuclei per unit tumor area, P≤0.001) in AO. While in most areas these changes were evident as typical microvascular proliferation, other areas showed thin walled vessels with increased MIB-1 positivity. VEGF was only assessed morphologically and showed positive staining of vasculature only, in WDO, while AO also showed immunoreactivity of vessels and multiple areas of tumor cells. These findings support a contributory role for vascular proliferation in assessing histologic grade. These findings also suggest that VEGF expression which is confined to blood vessels in lower grade tumors but eventually is expressed by tumor cells in higher grade oligodendrogliomas may be an important factor as the tumor progresses.


Application of Lean Principles to Neurosurgical Procedures: The Case of Lumbar Spinal Fusion Surgery, a Literature Review and Pilot Series.
Delivery of higher value healthcare is an ultimate government and public goal. Improving efficiency by standardization of surgical steps can improve patient outcomes, reduce costs, and lead to higher value healthcare. Lean principles and methodology have improved timeliness in perioperative medicine; however, process mapping of surgery itself has not been performed. To apply Plan/Do/Study/Act (PDSA) cycles methodology to lumbar posterior instrumented fusion (PIF) using lean principles to create a standard work flow, identify waste, remove intraoperative variability, and examine feasibility among pilot cases. Process maps for 5 PIF procedures were created by a PDSA cycle from 1 faculty neurosurgeon at 1 institution. Plan, modularize PIF into basic components; Do, map and time components; Study, analyze results; and Act, identify waste. Waste inventories, spaghetti diagrams, and chartings of time spent per step were created. Procedural steps were broadly defined in order to compare steps despite the variability in PIF and were analyzed with box and whisker plots to evaluate variability. Temporal variabilities in duration of decompression vs closure and hardware vs closure were significantly different (P = .003). Variability in procedural step duration was smallest for closure and largest for exposure. Wastes including waiting and instrument defects accounted for 15% and 66% of all waste, respectively. This pilot series demonstrates that lean principles can standardize surgical workflows and identify waste. Though time and labor intensive, lean principles and PDSA methodology can be applied to operative steps, not just the perioperative period.


Assessment of Structural Changes in Translucency and Opacity of Tooth Enamel against a Direct Demineralization Process: An In Vitro Study.
The objective of this study was to assess in vitro the structural changes in translucency and opacity of tooth enamel following a direct demineralization process. This experimental in vitro study evaluated 45 thirds (cervical, middle, and occlusal) of the tooth enamel surface of premolar teeth extracted from young adults divided into three groups of 15 specimens each: Group 1 (solution based on calcium, phosphorus, and fluorine), Group 2 (orthophosphoric acid 37%), and control group (distilled water). All underwent optical macroscopic examination with ×3 magnification to determine the initial translucency according to the variation of the medium in their intercrystalline spaces, and Thylstrup and Fejerskov Index was used. The experimental groups were then subjected to an artificial caries process during which the specimens were placed in an inorganic and organic solution of calcium, phosphorus, and fluorine at 37°C for 90 days with the acidic solution at pH 5 and the neutral solution at pH 7. The control specimens were placed in distilled water. Finally, all the specimens were assessed by polarization microscopy. In relation to the occlusal third, the highest proportion in Groups 1 and 2 was in Grades 2 and 3 (80%). A significant association was only observed between the experimental groups in the degree of translucency in the occlusal third (P = 0.002), whereas no association was found in relation to the degree of opacity in the middle and cervical thirds in either study group (P > 0.05). The resistance of enamel hydroxyapatite crystals increases from occlusal to cervical due to the greater presence of aprismatic enamel in the cervical horizontal third.


Practical Use of Infliximab Concentration Monitoring in Pediatric Crohn Disease.
Therapeutic drug monitoring (TDM) that guides infliximab (IFX) intensification strategies has been shown to improve IFX efficacy. We conducted a review to evaluate the utility of TDM in the assessment and subsequent management of IFX loss of response in our pediatric population with Crohn disease (CD). Single-center retrospective study of patients with CD receiving IFX that had TDM from December 2009 to September 2013. We defined subtherapeutic trough as a drug level below the detection limit of the Prometheus enzyme-linked immunoabsorbant assay and Anser reference values (1.4 and 1 μg/mL, respectively) or a mid-interval level <12 μg/mL. One hundred ninety-one IFX concentration tests were performed on 72 patients with CD with loss of response to therapy as the primary indication (72%). 34% of all TDM were subtherapeutic. After initial TDM, 25 of the 72 patients received regimen intensification with 72% in clinical remission at 6 months. Including all of the TDM that resulted in IFX dose intensification, we found a significant improvement in 6-month remission rates whether intensification followed mid-interval (88% remission) or trough (56% remission) testing (P = 0.026). Antibody to infliximab was found in 14 patients with 5 occurring in the first year of therapy. Furthermore, 71% of patients with antibody to infliximab that were switched to an alternative anti-tumor necrosis factor achieved clinical remission at six months. In multivariable regression analysis, we found IFX dose (mg/kg), IFX dosing frequency (weeks), and the erythrocyte sedimentation rate at the previous infusion were significantly associated with the IFX concentration. TDM in our pediatric population with CD led to informed clinical decisions and improved rates of clinical remission.


Serotonergic modulation of dopamine measured with [11C]raclopride and PET in normal human subjects.
This study was undertaken to measure serotonergic modulation of dopamine in vivo by using positron emission tomography (PET), a radiotracer for the striatal dopamine D2 receptor ([11C]raclopride), and a pharmacologic challenge of the serotonin system (d,l-fenfluramine). Two PET studies using [11C]raclopride were performed in 11 normal male subjects before administration of the serotonin-releasing agent and reuptake inhibitor fenfluramine (60 mg p.o.) and 3 hours afterward. A graphical analysis method was used with the [11C]raclopride data to derive the distribution volume of D2 receptors. Plasma levels of fenfluramine, norfenfluramine, homovanillic acid (HVA), cortisol, and prolactin were determined. Levels of fenfluramine and prolactin were elevated 2 hours after fenfluramine administration and remained significantly elevated during the second scan, while levels of HVA and cortisol were not altered significantly during the time of scanning. A significant decrease in the specific binding (striatum) and the nonspecific binding subtracted from the specific binding (striatum minus cerebellum) of [11C]raclopride was observed. The rate of metabolism of [11C]raclopride and the nonspecific binding (cerebellum) were not significantly altered by the fenfluramine intervention. The observed decrease in [11C]raclopride binding is consistent with an increase in dopamine concentrations and with the ability of serotonin to stimulate dopamine activity. The ability to measure serotonergic modulation of dopamine in vivo may have implications for the study of etiologic and therapeutic mechanisms in schizophrenia, major depressive disorder, obsessive-compulsive disorder, and substance abuse.


Parental health limitations, caregiving and loneliness among women with widowed parents: longitudinal evidence from France.
We investigate how daughters' feelings of loneliness are impacted when widowed parents develop health limitations, and when daughters take on personal care tasks in response. Using longitudinal data from daughters of widowed parents drawn from the French Family and Intergenerational Relationships Study (ERFI, 1485 observations nested in 557 daughters), we assess (a) whether health limitations of widowed parents are associated with daughters' feelings of loneliness regardless of whether or not daughters provide personal care and (b) whether there is an effect of care provision on loneliness that cannot be explained by parental health limitations. Fixed effect regression analyses show that widowed parents' health limitations were associated with raised feelings of loneliness among their daughters. No significant additional effect of providing personal care to a widowed parent was found. Prior research on the impact of health limitations of older parents on the lives of their adult-children has focused mostly on issues related to informal caregiving. Our findings suggest that more attention to the psychosocial impact of parental health limitations-net of actual caregiving-on adult children's lives is warranted.


Imaging and analysis of perivascular nerves in human mesenteric and coronary arteries: a comparison between epi-fluorescence and confocal microscopy.
Perivascular nerves supplying human arteries can be visualised after immunohistochemical staining for a variety of markers. The pattern and density of perivascular nerves vary with region, age and disease. Quantification of the nerve plexus, which may be performed by image analysis, is a prerequisite to assess differences in nerve density. The use of epi-fluorescence microscopy (EFM) presents difficulties in visualising the nerve plexus in certain tissues, which can affect the reliability with which specific staining can be localised and distinguished from non-specific staining. In this study, confocal scanning laser microscopy (CSLM) was used in parallel with EFM, in order to compare images from both techniques. In a comparison of identical areas of nerve plexuses of human mesenteric and coronary arteries stained for protein gene product (PGP) 9.5 and imaged using CSLM and EFM, higher values for area percent (area occupied by nerves), and intercept density (ID/mm, which reflects the number of nerve bundles detected) were found in CSLM images. Similar comparisons of unmatched epi-fluorescence and confocal images from a group of 45 mesenteric arteries revealed no significant difference for area percent, but significantly higher values for ID/mm in CSLM images. These findings illustrate that the better image quality in CSLM influences image analysis and can be very useful in studies of dynamic changes in nerve plexuses. We recommend CSLM for tissues that suffer from high background staining, such as human mesenteric and coronary arteries.


Aggressive hydration with Lactated Ringer's solution as the prophylactic intervention for postendoscopic retrograde cholangiopancreatography pancreatitis: A randomized controlled double-blind clinical trial.
Pancreatitis is a serious complication of endoscopic retrograde cholangiopancreatography (ERCP) and may cause significant morbidity and even death. There is no effective prophylactic intervention for patients with average risk yet. This study aims to investigate preventive effect of aggressive hydration for post-ERCP pancreatitis. In a double-blind controlled setting, 150 patient were randomly assigned to receive either aggressive hydration with lactated Ringer's solution (3 mL/kg/h during ERCP, followed by a 20 mL/kg bolus and 3 mL/kg/h for 8 h after the procedure, n = 75) or standard amount of hydration (1.5 mL/kg/h during and for 8 h after ERCP, n = 75). Patients were observed for volume overload as well as pancreatic pain and serum levels of amylase at baseline and 2, 8, and 24 h after ERCP. Post-ERCP pancreatitis was defined as hyperamylasemia (level of amylase >300) and pancreatic pain during the 24 h follow-up. Hyperamylasemia and pancreatic pain were the secondary end points. Mean age of the patients was 50.8 ± 13.5 years. Most of the patients were female (66%). Pancreatitis developed in 21 patients, including 22.7% of patients receiving standard hydration and 5.3% patients receiving aggressive hydration (P = 0.002). Hyperamylasemia was detected in 44.0% of patients receiving standard hydration and 22.7% of patients aggressive hydration (P = 0.006). The pancreatic pain was reported by 5.3% of patients receiving aggressive hydration and 37.3% of patients receiving standard hydration (P ≤ 0.005). Aggressive hydration with lactated Ringer's solution may effectively prevent post-ERCP pancreatitis as well as hyperamylasemia and pancreatic pain in patients with average risk.


Investigation of the Safety of Focused Ultrasound-Induced Blood-Brain Barrier Opening in a Natural Canine Model of Aging.
Rationale: Ultrasound-mediated opening of the Blood-Brain Barrier(BBB) has shown exciting potential for the treatment of Alzheimer's disease(AD). Studies in transgenic mouse models have shown that this approach can reduce plaque pathology and improve spatial memory. Before clinical translation can occur the safety of the method needs to be tested in a larger brain that allows lower frequencies be used to treat larger tissue volumes, simulating clinical situations. Here we investigate the safety of opening the BBB in half of the brain in a large aged animal model with naturally occurring amyloid deposits. Methods: Aged dogs naturally accumulate plaques and show associated cognitive declines. Low-frequency ultrasound was used to open the BBB unilaterally in aged beagles (9-11yrs, n=10) in accordance with institutionally approved protocols. Animals received either a single treatment or four weekly treatments. Magnetic resonance imaging(MRI) was used to guide the treatments and assess the tissue effects. The animals underwent neurological testing during treatment follow-up, and a follow-up MRI exam 1 week following the final treatment. Results: The permeability of the BBB was successfully increased in all animals (mean enhancement: 19±11% relative to untreated hemisphere). There was a single adverse event in the chronic treatment group that resolved within 24 hrs. Follow-up MRI showed the BBB to be intact with no evidence of tissue damage in all animals. Histological analysis showed comparable levels of microhemorrhage between the treated and control hemispheres in the prefrontal cortex (single/repeat treatment: 1.0±1.4 vs 0.4±0.5/5.2±1.8 vs. 4.0±2.0). No significant differences were observed in beta-amyloid load (single/repeat: p=0.31/p=0.98) although 3/5 animals in each group showed lower Aβ loads in the treated hemisphere. Conclusion: Whole-hemisphere opening of the BBB was well tolerated in the aged large animal brain. The treatment volumes and frequencies used are clinically relevant and indicate safety for clinical translation. Further study is warranted to determine if FUS has positive effects on naturally occurring amyloid pathology.


Automated image processing as an analytical tool in cell cryopreservation for bioprocess development.
The continuous availability of cells with defined cell characteristics represents a crucial issue in the biopharmaceutical and cell therapy industry. Here, development of cell banks with a long-term stability is essential and ensured by a cryopreservation strategy. The strategy needs to be optimized for each cell application individually and usually comprises controlled freezing, storage at ultra-low temperature, and fast thawing of cells. This approach is implemented by the development of master and working cell banks. Currently, empirical cryopreservation strategy development is standard, but a knowledge-based approach would be highly advantageous. In this article, we report the development of a video-based tool for the characterisation of freezing and thawing behaviour in cryopreservation process to enable a more knowledge-based cryopreservation process development. A successful tool validation was performed with a model cryopreservation process for the β-cell line INS-1E. Performance was evaluated for two working volumes (1.0 mL and 2.0 mL), based on freezing-thawing rates (20 °C to - 80 °C) and cell recovery and increase of biomass, to determine tool flexibility and practicality. Evaluation confirmed flexibility by correctly identifying a delay in freezing and thawing for the larger working volume. Further more, a decrease in cell recovery from 0.94 (± 0.14) % using 1.0 mL working volume to 0.61 (± 0.05) % using a 2.0 mL working volume displays tool practicality. The video-based tool proposed in this study presents a powerful tool for cell-specific optimisation of cryopreservation protocols. This can facilitate faster and more knowledge-based cryopreservation process development In this study, a video-based analytical tool was developed for the characterisation of freezing and thawing behaviour in cryopreservation process development. Evaluation of the practicality and flexibility of the developed tool was done based on a scale-up case study with the cell line INS-1E. Here, the influence of sample working volume on process performance was investigated. Increasing the volume from 1to 2 mL led to a delay in freezing and thawing behaviour which caused cell recovery loss. We believe that the developed tool will facilitate more directed and systematic cryopreservation process development.


Potential trade-off between vocal ornamentation and spatial ability in a songbird.
Bird song is hypothesized to be a reliable indicator of cognition because it depends on brain structure and function. Song features have been found to correlate positively with measures of cognition, but the relationship between song and cognition is complicated because not all cognitive abilities are themselves positively correlated. If cognition is not a unitary trait, developmental constraints on brain growth could generate trade-offs between some aspects of cognition and song. To further clarify the relationship between song and cognition in song sparrows (Melospiza melodia), we examined repertoire size and performance on a spatial task. We found an inverse relationship between repertoire size and speed of spatial learning and suggest that a developmental trade-off between the hippocampus and song control nuclei could be responsible for this relationship. By attending to male song, females may learn about a suite of cognitive abilities; this study suggests that females may glean information about a male's cognitive weaknesses as well as his strengths.


Podocyte foot process effacement in postreperfusion allograft biopsies correlates with early recurrence of proteinuria in focal segmental glomerulosclerosis.
Focal segmental glomerulosclerosis (FSGS) is a relatively prevalent glomerular disorder that often progresses to end-stage renal disease. Thirty to 80% of kidney transplant (KT) recipients with FSGS will experience recurrence characterized by proteinuria and podocyte damage. We hypothesized that the degree of podocyte foot process (FP) effacement in postreperfusion transplant biopsies can be used to predict the development of clinical recurrence of FSGS. Nineteen pairs of pre- and postreperfusion biopsy specimens were studied. We evaluated the degree of FP effacement in postreperfusion KT biopsies by counting the number of widened FP per capillary loop. Early recurrence of FSGS was defined as development of nephrotic range proteinuria between days 3 and 30 posttransplant. Early recurrence occurred in 7 of 19 grafts (36.8%) at a mean of 4.29±1.89 days. The mean score of FP effacement in postreperfusion allograft biopsies was 0.72±0.31 and 1.35±0.63 in the nonrecurrent and recurrent group, respectively (P=0.039). There was an association between FP effacement and proteinuria (P = 0.04). The FP effacement score predicts early recurrence with a sensitivity of 71.4% and specificity of 91.7%. FP effacement can be observed within minutes after reperfusion in renal transplantation of recipients with FSGS that will ultimately develop recurrent FSGS. This suggests a key role for the podocyte injury in the pathogenesis of recurrent FSGS and further supports the presence of circulating factors causing FP effacement. The FP effacement score in the postreperfusion KT biopsy may become a useful predictive test if validated in larger studies.


Magnetic resonance urography: a new gold standard for the evaluation of solitary kidneys and renal buds?
We evaluated the diagnostic potential of magnetic resonance urography (MRU) in pediatric patients with a solitary kidney and/or dysplastic orthotopic or ectopic renal buds. We present our evolving experience with this imaging procedure. A total of 30 patients with a mean age of 43.8 months with a known or suspected solitary kidney with or without clinical symptoms underwent ultrasound of the urinary tract as well as voiding cystourethrogram, renal static dimercapto-succinic acid (DMSA) scintigraphy and MRU. The results of these diagnostic imaging studies were then compared with each other and set against the results of the final diagnosis established at surgery and by histology, if available. Dysplastic orthotopic or dystopic renal buds suspected on ultrasound in 11 of the 30 patients were confirmed in all 11 by MRU. While accompanying ectopic ureteral insertions were shown in only 3 patients by ultrasound, they were visualized in all 4 by MRU. The other 7 patients with dysplastic renal buds had orthotopic ureteral orifices. DMSA scintigraphy failed to detect 10 renal buds. The diagnostic value of MRU appears to be superior to that of other imaging tests, even to that of DMSA scintigraphy, which with voiding cystourethrography is currently considered the gold standard for evaluating sonographically suspected solitary kidneys with contralateral orthotopic or ectopic renal buds and ectopic ureteral insertion.


Groundwater vulnerability to climate change: A review of the assessment methodology.
Impacts of climate change on water resources, especially groundwater, can no longer be hidden. These impacts are further exacerbated under the integrated influence of climate variability, climate change and anthropogenic activities. The degree of impact varies according to geographical location and other factors leading systems and regions towards different levels of vulnerability. In the recent past, several attempts have been made in various regions across the globe to quantify the impacts and consequences of climate and non-climate factors in terms of vulnerability to groundwater resources. Firstly, this paper provides a structured review of the available literature, aiming to critically analyse and highlight the limitations and knowledge gaps involved in vulnerability (of groundwater to climate change) assessment methodologies. The effects of indicator choice and the importance of including composite indicators are then emphasised. A new integrated approach for the assessment of groundwater vulnerability to climate change is proposed to successfully address those limitations. This review concludes that the choice of indicator has a significant role in defining the reliability of computed results. The effect of an individual indicator is also apparent but the consideration of a combination (variety) of indicators may give more realistic results. Therefore, in future, depending upon the local conditions and scale of the study, indicators from various groups should be chosen. Furthermore, there are various assumptions involved in previous methodologies, which limit their scope by introducing uncertainty in the calculated results. These limitations can be overcome by implementing the proposed approach.


Phytocannabinoids: a unified critical inventory.
Covering up to January 2016Cannabis sativa L. is a prolific, but not exclusive, producer of a diverse group of isoprenylated resorcinyl polyketides collectively known as phytocannabinoids. The modular nature of the pathways that merge into the phytocannabinoid chemotype translates in differences in the nature of the resorcinyl side-chain and the degree of oligomerization of the isoprenyl residue, making the definition of phytocannabinoid elusive from a structural standpoint. A biogenetic definition is therefore proposed, splitting the phytocannabinoid chemotype into an alkyl- and a β-aralklyl version, and discussing the relationships between phytocannabinoids from different sources (higher plants, liverworts, fungi). The startling diversity of cannabis phytocannabinoids might be, at least in part, the result of non-enzymatic transformations induced by heat, light, and atmospheric oxygen on a limited set of major constituents (CBG, CBD, Δ9-THC and CBC and their corresponding acidic versions), whose degradation is detailed to emphasize this possibility. The diversity of metabotropic (cannabinoid receptors), ionotropic (thermos-TRPs), and transcription factors (PPARs) targeted by phytocannabinoids is discussed. The integrated inventory of these compounds and their biological macromolecular end-points highlights the opportunities that phytocannabinoids offer to access desirable drug-like space beyond the one associated to the narcotic target CB1.


Symmetry breaking and the turn-on fluorescence of small, highly strained carbon nanohoops.
[n]Cycloparaphenylenes, or "carbon nanohoops," are unique conjugated macrocycles with radially oriented π-systems similar to those in carbon nanotubes. The centrosymmetric nature and conformational rigidity of these molecules lead to unusual size-dependent photophysical characteristics. To investigate these effects further and expand the family of possible structures, a new class of related carbon nanohoops with broken symmetry is disclosed. In these structures, referred to as meta[n]cycloparaphenylenes, a single carbon-carbon bond is shifted by one position in order to break the centrosymmetric nature of the parent [n]cycloparaphenylenes. Advantageously, the symmetry breaking leads to bright emission in the smaller nanohoops, which are typically non-fluorescent due to optical selection rules. Moreover, this simple structural manipulation retains one of the most unique features of the nanohoop structures-size dependent emissive properties with relatively large extinction coefficients and quantum yields. Inspired by earlier theoretical work by Tretiak and co-workers, this joint synthetic, photophysical, and theoretical study provides further design principles to manipulate the optical properties of this growing class of molecules with radially oriented π-systems.


Neural correlates of stereotype application.
Recent research has focused on the disparate mechanisms that support the human ability to "mentalize" about the thoughts and feelings of others. One such process may rely on precompiled, semantic beliefs about the characteristics common to members of a social group, that is, on stereotypes; for example, judging that a woman may be more likely than a man to have certain interests or opinions. In the current study, we identified a pattern of neural activity associated with the use of stereotypes to judge another person's psychological characteristics. During fMRI scanning, participants mentalized about the likely responses of a female and male target to a series of questions, some of which were related to gender stereotypes (e.g., "enjoys shopping for new clothes"). Trials on which participants applied a stereotype were segregated from those on which participants avoided stereotype use. The BOLD response in an extensive region of the right frontal cortex differentiated stereotype-applied from -unapplied trials. Moreover, this neural difference was correlated with a behavioral index of gender associations-the Implicit Association Test-administered after scanning. Results suggest that stereotype application may draw on cognitive processes that more generally subserve semantic knowledge about categories.


The 2003 Nicolas Andry Award. Orthopaedic gene therapy.
We review progress in the field of orthopaedic gene therapy since the concept of using gene transfer to address orthopaedic problems was initiated approximately 15 years ago. The original target, arthritis, has been the subject of two successful Phase I clinical trials, and additional human studies are pending in rheumatoid arthritis and osteoarthritis. The repair of damaged musculoskeletal tissues also has proved to be a fruitful area of research, and impressive enhancement of bone healing has been achieved in preclinical models. Rapid progress also is being made in the use of gene transfer to improve cartilage repair, ligament healing, and restoration of various additional tissues, including tendon and meniscus. Other applications include intervertebral disc degeneration, aseptic loosening, osteoporosis, genetic diseases, and orthopaedic tumors. Of these various orthopaedic targets of gene therapy, tissue repair is likely to make the earliest clinical impact because it can be achieved with existing technology. Tissue repair may become one of the earliest clinical successes for gene therapy as a whole. Orthopaedics promises to be a leading discipline for the use of human gene therapy.


Mirror-touch synaesthesia: Difficulties inhibiting the other.
Individuals with mirror touch synaesthesia (MTS) experience touch on their own body when observing others being touched. A recent account proposes that such rare experiences could be linked to impairment in self-other representations. Here we tested participants with MTS on a battery of social cognition tests and found that compared to non-synaesthete controls, the MTS group showed impairment in imitation-inhibition but not in visual perspective taking or theory of mind. Although all of these socio-cognitive abilities rely on the control of self-other representations, they differ as to whether the self, or the other, should be preferentially represented. For imitation-inhibition, representations of the other should be inhibited and self-representations should be enhanced, whereas the opposite is true for visual perspective taking and theory of mind. These findings suggest that MTS is associated with a specific deficit in inhibiting representation of other individuals and shed light on the fractionability of processes underlying typical social cognition.


Evidence for two paths of alcohol use onset in adolescents.
Research is needed to identify risk factors specifically associated with the development of substance abuse. The current study explored the possibility that adolescents classified as having a problem behavior prone orientation (Type II) are predisposed to more rapid alcohol use onset compared to more normally socialized (Type I) adolescents. It was hypothesized that both types of adolescents would increase their alcohol use over time, but that problem behavior prone adolescents would increase their rates of alcohol consumption more rapidly than would normally socialized adolescents. Using ANCOVA (with baseline alcohol use as a covariate) and t tests (examining only nondrinkers at baseline), the hypotheses were strongly supported. Both Type I and Type II adolescents significantly increased their alcohol use over a one-year period. Type II adolescents, in comparison to Type I adolescents, had significantly higher alcohol use. The greater alcohol use among Type II adolescents was attributed to their problem behavior prone orientation. The findings suggest the existence of two different developmental pathways of alcohol use onset, one initiated by normally socialized adolescents and the other by adolescents with a problem behavior prone orientation. Future alcohol abuse prevention programs may benefit from tailored intervention strategies which take into account population specific risk factors.


What men who have sex with men think about the human papillomavirus vaccine.
This study aimed to ascertain the attitudes of men who have sex with men (MSM) to the human papillomavirus (HPV) vaccine and to determine the age at which MSM would be willing to ask for the HPV vaccine in relation to their age of sexual debut. Of 205 MSM attending the Melbourne Sexual Health Centre between December 2007 and January 2008, 200 (98%; median age 27 years) completed the study questionnaire. Only 30% were aware that there was a vaccine available for protection against infection with certain HPV types. When informed of the increased risk of anal cancer among MSM, 47% of MSM indicated that they would be willing to pay $A450 for the vaccine course. A total of 93% indicated that they would be willing to disclose that they were MSM to a health professional in order to obtain the vaccine for free, but not until a median age of 20 years: 2 years after the median age of sexual debut (18 years) and after a median of 15 sexual partners. If the HPV vaccine is targeted to MSM, the challenge will be for MSM to be vaccinated before they acquire HPV infection.


Stereoretention in styrene heterodimerisation promoted by one-electron oxidants.
Radical cations generated from the oxidation of C[double bond, length as m-dash]C π-bonds are synthetically useful reactive intermediates for C-C and C-X bond formation. Radical cation formation, induced by sub-stoichiometric amounts of external oxidant, are important intermediates in the Woodward-Hoffmann thermally disallowed [2 + 2] cycloaddition of electron-rich alkenes. Using density functional theory (DFT), we report the detailed mechanisms underlying the intermolecular heterodimerisation of anethole and β-methylstyrene to give unsymmetrical, tetra-substituted cyclobutanes. Reactions between trans-alkenes favour the all-trans adduct, resulting from a kinetic preference for anti-addition reinforced by reversibility at ambient temperatures since this is also the thermodynamic product; on the other hand, reactions between a trans-alkene and a cis-alkene favour syn-addition, while exocyclic rotation in the acyclic radical cation intermediate is also possible since C-C forming barriers are higher. Computations are consistent with the experimental observation that hexafluoroisopropanol (HFIP) is a better solvent than acetonitrile, in part due to its ability to stabilise the reduced form of the hypervalent iodine initiator by hydrogen bonding, but also through the stabilisation of radical cationic intermediates along the reaction coordinate.