Desensitization of adenosine and dopamine receptors in rat brain after treatment with adenosine analogs.
Maximally tolerated doses of N6-[(R)-1-methyl-2-phenylethyl] adenosine (0.50 nmol/hr/2 wk), 5'-N-ethylcarboxamide adenosine (NECA, 0.04 nmol/hr/2 wk) or deoxycoformycin (5 nmol/hr/1 wk) were administered i.c.v. to rats using mini-osmotic pumps. Adenosine receptor function was subsequently assayed using both ligand binding and adenylate cyclase assays. Binding to A1 receptors was quantitated using [3H]N6-[(R)-1-methyl-2-phenylethyl]adenosine, a selective agonist ligand at A1 receptors. Differences in the binding of this ligand and that of [3H]NECA, which binds to A1 and A2 receptors with similar affinities, were used to quantitate A2 receptors. None of the treatments affected A1 receptor function as assessed by both ligand binding and adenylate cyclase assays. A2 receptor binding and A2 receptor-mediated stimulation of adenylate cyclase were blunted in striatal membranes from NECA- and deoxycoformycin-treated rats but unaffected in striatal membranes from N6-[(R]-1-methyl-2-phenylethyl]adenosine-treated rats. All three pretreatments attenuated D1 dopamine receptor-mediated stimulation of adenylate cyclase in striatal membranes. These results suggest that 1) the A2 adenosine receptor system is susceptible to desensitization and 2) different mechanisms are involved in the NECA- and deoxycoformycin-induced desensitization of A2 adenosine receptor and D1 dopamine receptor systems. It is suggested that the D1 dopamine receptor desensitization is, in fact, due to the tonic stimulation of adenosine A1 receptors.


A versatile clearing agent for multi-modal brain imaging.
Extensive mapping of neuronal connections in the central nervous system requires high-throughput µm-scale imaging of large volumes. In recent years, different approaches have been developed to overcome the limitations due to tissue light scattering. These methods are generally developed to improve the performance of a specific imaging modality, thus limiting comprehensive neuroanatomical exploration by multi-modal optical techniques. Here, we introduce a versatile brain clearing agent (2,2'-thiodiethanol; TDE) suitable for various applications and imaging techniques. TDE is cost-efficient, water-soluble and low-viscous and, more importantly, it preserves fluorescence, is compatible with immunostaining and does not cause deformations at sub-cellular level. We demonstrate the effectiveness of this method in different applications: in fixed samples by imaging a whole mouse hippocampus with serial two-photon tomography; in combination with CLARITY by reconstructing an entire mouse brain with light sheet microscopy and in translational research by imaging immunostained human dysplastic brain tissue.


Intraoperative radiotherapy in the multidisciplinary treatment of pediatric tumors. A preliminary report on initial results.
From September 1984 to July 1987, 33 children received intraoperative radiotherapy as part of a multidisciplinary tumor treatment. Their age ranged from 2 to 17 years. Tumors types: Ewing's sarcoma (n = 11), osteosarcoma (n = 8), soft tissue sarcomas (n = 5), Wilms' tumor (n = 3), neuroblastoma (n = 3), malignant pheochromocytoma (n = 1), Hodgkin's disease (n = 1), and optic nerve glioma (n = 1). In 25 patients the disease was localized while 8 had distant metastases. Intraoperative radiotherapy was used in 26 previously untreated patients as part of a radical treatment program and in 7 cases as an effort to rescue local failures (5 in previously irradiated areas). The intraoperative radiation field included the surgically exposed tumor or tumor bed, and the single doses ranged from 10 to 20 Gy, with 6-20 MeV electrons. Patients with osteosarcoma and recurrent tumor in a previously irradiated area did not receive postoperative external beam radiotherapy. With a median follow-up time of 10 months (1 to 31 + months) 24 out of 33 patients are alive without local recurrence and 9 have died from tumor (5 with local disease progression). Intraoperative radiotherapy seems to be a feasible treatment which might promote local control in pediatric tumors.


Mesozoic origin for West Indian insectivores.
The highly endangered solenodons, endemic to Cuba (Solenodon cubanus) and Hispaniola (S. paradoxus), comprise the only two surviving species of West Indian insectivores. Combined gene sequences (13.9 kilobases) from S. paradoxus established that solenodons diverged from other eulipotyphlan insectivores 76 million years ago in the Cretaceous period, which is consistent with vicariance, though also compatible with dispersal. A sequence of 1.6 kilobases of mitochondrial DNA from S. cubanus indicated a deep divergence of 25 million years versus the congeneric S. paradoxus, which is consistent with vicariant origins as tectonic forces separated Cuba and Hispaniola. Efforts to prevent extinction of the two surviving solenodon species would conserve an entire lineage as old or older than many mammalian orders.


Aneurysmal and haemangiopericytoma-like fibrous histiocytoma.
To describe the clinicopathological features of 33 aneurysmal fibrous histiocytomas (AFH), including five cases with a haemangiopericytoma-like pattern. Thirty three cases of AFH were studied by using routine histology and immunohistochemistry for factor XIIIa, the "cell activity marker" E9 (anti-metallothionein), NK1C3 (CD57), smooth muscle actin (SMA), factor VIII, ulex europaeus agglutinin, JC70A (CD31), and QBEND10 (CD34). The time dependent variation in histopathological features was evaluated by statistical methods (Pearson chi 2, likelihood ratio chi 2). Of the AFHs, 29 of 33 occurred on the extremities of adults (age range 30 to 50 years), six of which were associated with rapid growth, probably caused by trauma, and pain. Twenty one lesions were thought to be vascular and/or melanocytic lesions, including two melanomas, because of a bluish-black and/or cystic appearance. Histologically, large areas of haemorrhage, up to 50% of the tumour bulk, lacking an endothelial lining were seen in otherwise typical fibrous histiocytomas. Five cases resembled nodular stages of Kaposi's sarcoma. Variable haemosiderin deposition in histiocytes (18/33) and giant cells (11/33) was suggestive of haemosiderotic histiocytoma. A haemangiopericytoma-like pattern was seen in five otherwise indistinguishable cases. On immunohistochemistry, variable reactivity was seen for factor XIIIa (18/30), with E9 (18/30), NK1C3 (19/30), and for SMA (14/30), but labelling for vascular markers was not detected. Early lesions without iron deposition were factor XIIIa positive; late lesions with iron deposition were factor XIIIa negative. Labelling for SMA correlated with prominent sclerosis. AFHs, including a haemangiopericytoma-like variant, have a characteristic time dependent histological and immunophenotypic profile, clearly different from nodular type Kaposi's sarcoma.


Reduced neutrophil count in people of African descent is due to a regulatory variant in the Duffy antigen receptor for chemokines gene.
Persistently low white blood cell count (WBC) and neutrophil count is a well-described phenomenon in persons of African ancestry, whose etiology remains unknown. We recently used admixture mapping to identify an approximately 1-megabase region on chromosome 1, where ancestry status (African or European) almost entirely accounted for the difference in WBC between African Americans and European Americans. To identify the specific genetic change responsible for this association, we analyzed genotype and phenotype data from 6,005 African Americans from the Jackson Heart Study (JHS), the Health, Aging and Body Composition (Health ABC) Study, and the Atherosclerosis Risk in Communities (ARIC) Study. We demonstrate that the causal variant must be at least 91% different in frequency between West Africans and European Americans. An excellent candidate is the Duffy Null polymorphism (SNP rs2814778 at chromosome 1q23.2), which is the only polymorphism in the region known to be so differentiated in frequency and is already known to protect against Plasmodium vivax malaria. We confirm that rs2814778 is predictive of WBC and neutrophil count in African Americans above beyond the previously described admixture association (P = 3.8 x 10(-5)), establishing a novel phenotype for this genetic variant.


Responding to global stimulant use: challenges and opportunities.
We did a global review to synthesise data on the prevalence, harms, and interventions for stimulant use, focusing specifically on the use of cocaine and amphetamines. Modelling estimated the effect of cocaine and amphetamine use on mortality, suicidality, and blood borne virus incidence. The estimated global prevalence of cocaine use was 0·4% and amphetamine use was 0·7%, with dependence affecting 16% of people who used cocaine and 11% of those who used amphetamine. Stimulant use was associated with elevated mortality, increased incidence of HIV and hepatitis C infection, poor mental health (suicidality, psychosis, depression, and violence), and increased risk of cardiovascular events. No effective pharmacotherapies are available that reduce stimulant use, and the available psychosocial interventions (except for contingency management) had a weak overall effect. Generic approaches can address mental health and blood borne virus infection risk if better tailored to mitigate the harms associated with stimulant use. Substantial and sustained investment is needed to develop more effective interventions to reduce stimulant use.


Thermal conductivities of some novel nonlinear optical materials.
Results of thermal conductivity measurements are reported for several of the more recently developed nonlinear optical crystals. New or substantially revised values of thermal conductivity were obtained in six materials. Notable thermal conductivities measured were those for AgGaS(2) [0.014 W/(cm K) and 0.015 W/(cm K)], AgGaSe(2) [0.010 W/(cm K) and 0.011 W/(cm K)], beta barium borate [0.016 W/(cm K) and 0.012 W/(cm K)], and ZnGeP(2) [0.36 W/(cm K) and 0.35 W/(cm K)], with values quoted for directions respectively parallel and perpendicular to the optic axis for each material. These new data provide necessary input for the design of high-power optical frequency converters.


Predictive factors of [(11)C]choline PET/CT in patients with biochemical failure after radical prostatectomy.
Detection of recurrence in prostate cancer patients with biochemical failure after radical prostatectomy by [(11)C]choline PET/CT depends on the prostate-specific antigen (PSA) level. The role of other clinical and pathological variables has not been explored. A total of 2,124 prostate cancer patients referred to our Institution for [(11)C]choline PET/CT from December 2004 to January 2007 for restaging of disease were retrospectively considered for this study. Inclusion criteria were: previous treatment by radical prostatectomy, and biochemical failure, defined as at least two consecutive PSA measurements of >0.2 ng/ml. These criteria were met for 358 patients. Binary logistic analysis was used to investigate the predictive factors of [(11)C]choline PET/CT. PET/CT findings were validated using criteria based on histological analysis, and follow-up clinical and imaging data. Receiver operating characteristic (ROC) analysis was used to assess the performance of [(11)C]choline PET/CT in relation to PSA levels. The mean PSA level was 3.77 +/- 6.94 ng/ml (range 0.23-45 ng/ml; median 1.27 ng/ml). PET/CT was positive for recurrence in 161 of 358 patients (45%). On an anatomical region basis, [(11)C]choline pathological uptake was observed in lymph nodes (107/161 patients, 66%), prostatectomy bed (55/161 patients, 34%), and in the skeleton (46/161 patients, 29%). PET/CT findings were validated using histological criteria (46/358, 13%), and follow-up clinical and imaging criteria (312/358, 87%). Sensitivity, specificity, positive predictive value, negative predictive value, and overall accuracy were, respectively, 85%, 93%, 91%, 87%, and 89%. In multivariate analysis, high PSA levels, advanced pathological stage, previous biochemical failure and older age were significantly (P < 0.05) associated with an increased risk of positive PET/CT findings. The percentage of positive scans was 19% in those with a PSA level between 0.2 and 1 ng/ml, 46% in those with a PSA level between 1 and 3 ng/ml, and 82% in those with a PSA level higher than 3 ng/ml. ROC analysis showed that PET/CT-positive and PET/CT-negative patients could be best distinguished using a PSA cut-off value of 1.4 ng/ml. In addition to PSA levels, pathological stage, previous biochemical failure and age should be considered by physicians when referring prostate cancer patients with biochemical failure after radical prostatectomy to [(11)C]choline PET/CT.


Attention trajectories, mechanisms and outcomes: at the interface between developing cognition and environment.
Attentional processes play a crucial role in prioritizing information for further processing and they therefore sit at the interface between internal goals and the challenges presented by the environment. How does attentional control interact with the changing constraints imposed by the developing cognitive system? Emerging work in this area has employed a range of complementary techniques, from increasingly refined neurocognitive measures in typically developing individuals, to the investigation of risk or protective factors influencing attention trajectories in developmental disorders. A growing corpus of data suggests that, while attentional biases for specific input characteristics (e.g. suddenly appearing stimuli, emotional expressions) are in place from infancy, it is the interplay between these predispositions, genetic and environmental factors that drives attention development over time. With the advent of multidisciplinary approaches to the developmental cognitive neuroscience of attention, unravelling these complex dynamics from infancy and their outcome on learning is increasingly within reach.


A new spreadsheet method for the analysis of bivariate flow cytometric data.
A useful application of flow cytometry is the investigation of cell receptor-ligand interactions. However such analyses are often compromised due to problems interpreting changes in ligand binding where the receptor expression is not constant. Commonly, problems are encountered due to cell treatments resulting in altered receptor expression levels, or when cell lines expressing a transfected receptor with variable expression are being compared. To overcome this limitation we have developed a Microsoft Excel spreadsheet that aims to automatically and effectively simplify flow cytometric data and perform statistical tests in order to provide a clearer graphical representation of results. To demonstrate the use and advantages of this new spreadsheet method we have investigated the binding of the transmembrane adhesion receptor CD44 to its ligand hyaluronan. In the first example, phorbol ester treatment of cells results in both increased CD44 expression and increased hyaluronan binding. By applying the spreadsheet method we effectively demonstrate that this increased ligand binding results from receptor activation. In the second example we have compared AKR1 cells transfected either with wild type CD44 (WT CD44) or a mutant with a truncated cytoplasmic domain (CD44-T). These two populations do not have equivalent receptor expression levels but by using the spreadsheet method hyaluronan binding could be compared without the need to generate single cell clones or FACS sorting the cells for matching CD44 expression. By this method it was demonstrated that hyaluronan binding requires a threshold expression of CD44 and that this threshold is higher for CD44-T. However, at high CD44-T expression, binding was equivalent to WT CD44 indicating that the cytoplasmic domain has a role in presenting the receptor at the cell surface in a form required for efficient hyaluronan binding rather than modulating receptor activity. Using the attached spreadsheets and instructions, a simple post-acquisition method for analysing bivariate flow cytometry data is provided. This method constitutes a straightforward improvement over the standard graphical output of flow cytometric data and has the significant advantage that ligand binding can be compared between cell populations irrespective of receptor expression levels.


[Mortality risk factors in patients over 65 years of age hospitalized in a São Paulo, Brasil university hospital].
Various studies show that age in itself is not an independent predictive factor for the survival of critically ill older persons. In spite of that fact, people over 65 tend not to have access to the most-sophisticated diagnostic and therapeutic resources. With the continued aging of the population, it is increasingly important to be able to determine the mortality risk factors that affect the survival possibilities of hospitalized older people, especially in emergency situations. The objective of this study was to analyze the risk factors related to mortality among people over 65 admitted to the emergency service of the Santa Casa Hospital of São Paulo, a third-level academic institution. The study was based on a nonconditional logistic regression analysis of personal data in the clinical histories and other documents of the hospitalized patients. From July 1993 through March 1994, 599 patients over 65 were hospitalized. They included 326 men (54.4%) and 273 women (45.6%), with a median age of 73.3 years. The primary reasons for admission were pneumonia (14.4%), cerebrovascular disease (11.5%), and congestive heart failure (8.2%). There were 160 deaths among the group. Those who died had a median stay in the hospital of 4 days (range, 1 to 72), which was similar to that of those who survived (median, 3 days; range, 0 to 35 days; P = 0.29). According to multivariate analysis, factors predictive of survival were hypertension (odds ratio = 0.39; 95% CI: 0.23 to 0.68), chronic obstructive pulmonary disease (OR = 0.45; 95% CI: 0.22 to 0.95), and diabetes mellitus (OR = 0.50; 95% CI: 0.27 to 0.91). This was true regardless of sex, age, race, compliance with treatment, initial diagnosis, and other pathological conditions present. On the other hand, two factors were predictive of mortality during hospitalization. They were the presence of extrapulmonary infections (OR = 2.34; 95% CI: 1.13 to 4.86) and the number of preexisting illnesses: one (OR = 2.78; 95% CI: 1.56 to 4.96), two (OR = 4.56; 95% CI: 2.28 to 9.15), and three or more (OR = 15.88; 95% CI: 6.49 to 38.85). This study shows that improving the diagnosis and treatment of infections can reduce mortality of elderly persons admitted to emergency services. Rather than age, the multiplicity of diseases was the factor that increased the risk of death among these patients. There is no justification for using age alone to limit the access that older patients have to better diagnosis and treatment resources, if these resources can result in better survival rates, fewer disabilities, or a better quality of life.


pHMA, a pH-sensitive GFP reporter for cell engulfment, in Drosophila embryos, tissues, and cells.
Engulfment of apoptotic cells by phagocytosis ensures the removal of unwanted and defective cells. We developed a genetically encoded marker for cell engulfment, pHMA, which consists of the pH-Sensitive derivative of GFP, pHluorin, fused to the actin-binding domain of Moesin. In healthy cells of Drosophila embryos and cultured cells, pHMA resides at the cell cortex. In dying cells, pHMA loses its cortical localization and reports a modest decrease in pH. In embryos, the dying cells lose their apical contacts, then move basally and are ultimately engulfed by neighboring cells or macrophages. The cell corpse material is strongly acidified soon after engulfment and persists in the phagocytic cell for several hours. Changes in the pHMA signal correlate well with increases or decreases in apoptosis. These data show that pHMA is a useful reporter for cell engulfment and can be used in screening for mutations that affect cell engulfment.


Angiographic, hemodynamic and histological characterization of an arteriovenous fistula in rats.
Our understanding of the pathogenesis of arteriovenous malformations (AVMs) and arteriovenous fistulas (AVFs) has been limited by the lack of adequate animal models. In this study we evaluate the time course of angiographic, hemodynamic and histopathological changes in an arteriovenous fistula in rats as a potential model. An arteriovenous fistula was created by a side-to-end anastomosis of the common carotid artery (CCA) to the external jugular vein (EJV). The animals underwent angiography of the fistula and were sacrificed 1, 7, 21, 42 or 90 days later. Flow and pressure measurements were performed in the CCA and ipsi- and contralateral EJV and detailed histological examination of whole mount sections of the fistula and cranium were done on fixed sections. Immunohistochemistry for CD31, smooth muscle alpha-actin and Ki-67 were performed. Hemodynamic changes occur immediately after fistula formation creating a stable high flow, low resistant state. This induces a gradual increase in the inner diameter of the EJV and transverse sinus followed by a decrease in size of the transverse sinus. This decrease is associated with increased expression of alpha-actin in the wall of the sinus. The fistula becomes angiographically and histologically stable after 21 days. This model describes the time course of hemodynamic and histopathological changes after occur after AVF formation. Stabilization after 21 days makes it an attractive model for mechanistic and therapeutic studies of AVFs.


A constant flux of diverse thermophilic bacteria into the cold Arctic seabed.
Microorganisms have been repeatedly discovered in environments that do not support their metabolic activity. Identifying and quantifying these misplaced organisms can reveal dispersal mechanisms that shape natural microbial diversity. Using endospore germination experiments, we estimated a stable supply of thermophilic bacteria into permanently cold Arctic marine sediment at a rate exceeding 10(8) spores per square meter per year. These metabolically and phylogenetically diverse Firmicutes show no detectable activity at cold in situ temperatures but rapidly mineralize organic matter by hydrolysis, fermentation, and sulfate reduction upon induction at 50 degrees C. The closest relatives to these bacteria come from warm subsurface petroleum reservoir and ocean crust ecosystems, suggesting that seabed fluid flow from these environments is delivering thermophiles to the cold ocean. These transport pathways may broadly influence microbial community composition in the marine environment.


Is vitellogenin an ancestor of apolipoprotein B-100 of human low-density lipoprotein and human lipoprotein lipase?
Vitellogenin, an ancient animal protein, is the major yolk protein of eggs, where it is used as a food source during embryogenesis. Here it is shown that vitellogenins, including those from the invertebrates Caenorhabditis elegans and Drosophila melanogaster, contain domains that are homologous with parts of apolipoprotein B-100 (apoB-100) of human low-density lipoprotein and human lipoprotein lipase. As vitellogenins are likely to have been used by invertebrates during embryogenesis well before the circulation of lipids appeared in vertebrates, it is suggested that copies of a precursor gene, serving a function similar to vitellogenin, were modified to code for part of apoB-100 and lipoprotein lipase in vertebrates. In addition to providing a link between invertebrates and vertebrates for proteins involved in lipid transport, these homologies suggest new functions for vitellogenin other than being a yolk food for the developing embryo.


Confronting the issues of patient safety and investigator conflict of interest in an international clinical trial of myocardial reperfusion. Global Utilization of Streptokinase and Tissue Plasminogen Activator for Occluded Coronary Arteries (GUSTO) Steering Committee.
The Global Utilization of Streptokinase and Tissue Plasminogen Activator for Occluded Coronary Arteries (GUSTO) trial is a large scale international trial of new myocardial reperfusion strategies. The primary hypothesis is that early and sustained coronary artery recanalization will be associated with a significant reduction in mortality. The four regimens that are being tested are 1) streptokinase with subcutaneous heparin; 2) streptokinase with intravenous heparin; 3) accelerated recombinant tissue-type plasminogen activator (rt-PA) with intravenous heparin; and 4) combination streptokinase, rt-PA and intravenous heparin. The planned recruitment of 41,600 patients in 1,500 sites from 15 countries is expected to be completed by December 1992 and will enable detection of a 15% reduction or 1% absolute difference in mortality compared with that associated with standard therapy (streptokinase and subcutaneous heparin). In designing the trial, two important issues were directly addressed. First, a strategy was developed to provide assurance of patient safety during large scale investigational use of an aggressive thrombolytic regimen. This includes fascimile transmission of a one-page safety summary form to the Data Coordinating Center within 24 h of death or discharge, acceptance of the concept of "net clinical benefit" and close surveillance of the trial's progress by the independent Data and Safety Monitoring Committee. Second, to avoid potential conflict of interest beyond elimination of any position of financial equity, the Steering Committee unanimously voted to prohibit any honoraria for speaking engagements, payment for consultancy or travel or reimbursement of any kind from any of the five corporate sponsors until 1 year after publication of the results. Incorporation of these approaches may facilitate the design of future large scale randomized trials in cardiovascular medicine.


Transmission of an inhomogeneous plane wave through an electrically small aperture in a perfectly conducting plane screen.
Most solutions for electromagnetic diffraction by a circular aperture in a perfectly conducting plane screen are for an incident homogeneous (propagating) plane wave. When the aperture is electrically small (dimensions small compared to the wavelength), the well-known transmission coefficient behaves as the fourth power of the diameter/wavelength. We consider the case in which the incident field is an inhomogeneous (evanescent) plane wave. Numerical calculations for the electrically small circular aperture show that the transmission coefficient for an inhomogeneous plane wave can be substantially greater than for a homogeneous plane wave at the same frequency. This observation may be helpful in explaining the increased transmission recently reported for electrically small apertures in plane screens with modifications. The numerical calculations for the electrically small aperture are in agreement with results from approximate analytical expressions that are based on the equivalent electric and magnetic dipole moments for the electrically small complementary disk.


Habitual Caffeine Consumption Does Not Affect the Ergogenicity of Coffee Ingestion During a 5 km Cycling Time Trial.
There is growing evidence that caffeine and coffee ingestion prior to exercise provide similar ergogenic benefits. However, there has been a long-standing paradigm that habitual caffeine intake may influence the ergogenicity of caffeine supplementation. The aim of the present study was to investigate the effect of habitual caffeine intake on 5-km cycling time-trial performance following the ingestion of caffeinated coffee. Following institutional ethical approval, in a double-blind, randomized, crossover, placebo-controlled design, 46 recreationally active participants (27 men and 19 women) completed a 5-km cycling time trial on a cycle ergometer 60 m in following the ingestion of 0.09 g/kg coffee providing 3 mg/kg of caffeine, or a placebo. Habitual caffeine consumption was assessed using a caffeine consumption questionnaire with low habitual caffeine consumption defined as <3 and ≥6 mg · kg-1 · day-1 defined as high. An analysis of covariance using habitual caffeine intake as a covariant was performed to establish if habitual caffeine consumption had an impact on the ergogenic effect of coffee ingestion. Sixteen participants were classified as high-caffeine users and 30 as low. Ingesting caffeinated coffee improved 5-km cycling time-trial performance by 8 ± 12 s; 95% confidence interval (CI) [5, 13]; p < .001; d = 0.30, with low, 9±14 s; 95% CI [3, 14]; p = .002; d = 0.18, and high, 8 ± 10 s; 95% CI [-1, 17]; p = .008; d = 0.06, users improving by a similar magnitude, 95% CI [-12, 12]; p = .946; d = 0.08. In conclusion, habitual caffeine consumption did not affect the ergogenicity of coffee ingestion prior to a 5-km cycling time trial.


Deficits in facial, body movement and vocal emotional processing in autism spectrum disorders.
Previous behavioural and neuroimaging studies of emotion processing in autistic spectrum disorder (ASD) have focused on the use of facial stimuli. To date, however, no studies have examined emotion processing in autism across a broad range of social signals. This study addressed this issue by investigating emotion processing in a group of 23 adults with ASD and 23 age- and gender-matched controls. Recognition of basic emotions ('happiness', 'sadness', 'anger', disgust' and 'fear') was assessed from facial, body movement and vocal stimuli. The ability to make social judgements (such as approachability) from facial stimuli was also investigated. Significant deficits in emotion recognition were found in the ASD group relative to the control group across all stimulus domains (faces, body movements and voices). These deficits were seen across a range of emotions. The ASD group were also impaired in making social judgements compared to the control group and this correlated with impairments in basic emotion recognition. This study demonstrates that there are significant and broad-ranging deficits in emotion processing in ASD present across a range of stimulus domains and in the auditory and visual modality; they cannot therefore be accounted for simply in terms of impairments in face processing or in the visual modality alone. These results identify a core deficit affecting the processing of a wide range of emotional information in ASD, which contributes to the impairments in social function seen in people with this condition.


Does simultaneous bilingualism aggravate children's specific language problems?
There is little data on whether or not a bilingual upbringing may aggravate specific language problems in children. This study analysed whether there was an interaction of such problems and simultaneous bilingualism. Participants were 5- to 7-year-old children with specific language problems (LANG group, N = 56) or who were typically developing (CONTR group, N = 60). Seventy-three children were Swedish-Finnish bilingual and 43 were Swedish-speaking monolingual. Assessments (in Swedish) included tests of expressive language, comprehension, repetition and verbal memory. Per definition, the LANG group had lower scores than the CONTR group on all language tests. The bilingual group had lower scores than the monolingual group only on a test of body part naming. Importantly, the interaction of group (LANG or CONTR) and bilingualism was not significant on any of the language scores. Simultaneous bilingualism does not aggravate specific language problems but may result in a slower development of vocabulary both in children with and without specific language problems. Considering also advantages, a bilingual upbringing is an option also for children with specific language problems. In assessment, tests of vocabulary may be sensitive to bilingualism, instead tests assessing comprehension, syntax and nonword repetition may provide less biased methods.


Aberrant fucosylation enables breast cancer clusterin to interact with dendritic cell-specific ICAM-grabbing non-integrin (DC-SIGN).
Clusterin is a glycoprotein able to mediate different physiological functions such as control of complement activation, promotion of unfolded protein clearance and modulation of cell survival. Clusterin is overexpressed in many types of cancers and a large body of evidence suggests that it promotes carcinogenesis and tumor progression. We have previously described a novel clusterin glycoform present in human semen, but not in serum, highly enriched in terminal fucose motifs. Here we show that human luminal breast cancer (LBC) clusterin also bears terminal fucosylated glycans, conferring clusterin the ability to interact with DC-SIGN, a C-type lectin receptor expressed by myeloid cells. This clusterin glycosylation pattern was absent or diminished in non-involved juxtatumoral tissue, suggesting that fucosylated clusterin might represent a cancer associated glycoform. We also found that DC-SIGN is expressed by luminal breast cancer intratumoral macrophages. Moreover, experiments performed in vitro using semen fucosylated clusterin and monocyte derived macrophages showed that the interaction of semen clusterin with DC-SIGN promoted a proangiogenic profile, characterized by a high production of VEGF, IL-8 and TNF-α. Our results reveal an unexpected complexity on the structure and function of secretory clusterin produced by tumors and suggest that fucosylated clusterin produced by luminal breast cancer cells might play a role in tumor progression by promoting the release of pro-angiogenic factors by intratumoral macrophages.


Synaptic transmission parallels neuromodulation in a central food-intake circuit.
NeuromedinU is a potent regulator of food intake and activity in mammals. In Drosophila, neurons producing the homologous neuropeptide hugin regulate feeding and locomotion in a similar manner. Here, we use EM-based reconstruction to generate the entire connectome of hugin-producing neurons in the Drosophila larval CNS. We demonstrate that hugin neurons use synaptic transmission in addition to peptidergic neuromodulation and identify acetylcholine as a key transmitter. Hugin neuropeptide and acetylcholine are both necessary for the regulatory effect on feeding. We further show that subtypes of hugin neurons connect chemosensory to endocrine system by combinations of synaptic and peptide-receptor connections. Targets include endocrine neurons producing DH44, a CRH-like peptide, and insulin-like peptides. Homologs of these peptides are likewise downstream of neuromedinU, revealing striking parallels in flies and mammals. We propose that hugin neurons are part of an ancient physiological control system that has been conserved at functional and molecular level.


The melanin-concentrating hormone receptor 1, a novel target of autoantibody responses in vitiligo.
Vitiligo is a common depigmenting disorder resulting from the loss of melanocytes in the skin. The pathogenesis of the disease remains obscure, although autoimmune mechanisms are thought to be involved. Indeed, autoantibodies and autoreactive T lymphocytes that target melanocytes have been reported in some vitiligo patients. The objective of this study was to identify pigment cell antigens that are recognized by autoantibodies in vitiligo. Using IgG from vitiligo patients to screen a melanocyte cDNA phage-display library, we identified the melanin-concentrating hormone receptor 1 (MCHR1) as a novel autoantigen related to this disorder. Immunoreactivity against the receptor was demonstrated in vitiligo patient sera by using radiobinding assays. Among sera from healthy controls and from patients with autoimmune disease, none exhibited immunoreactivity to MCHR1, indicating a high disease specificity for Ab's against the receptor. Inhibition of MCH binding to its receptor by IgG from vitiligo patients was also shown.


In vivo function of Hsp90 is dependent on ATP binding and ATP hydrolysis.
Heat shock protein 90 (Hsp90), an abundant molecular chaperone in the eukaryotic cytosol, is involved in the folding of a set of cell regulatory proteins and in the re-folding of stress-denatured polypeptides. The basic mechanism of action of Hsp90 is not yet understood. In particular, it has been debated whether Hsp90 function is ATP dependent. A recent crystal structure of the NH2-terminal domain of yeast Hsp90 established the presence of a conserved nucleotide binding site that is identical with the binding site of geldanamycin, a specific inhibitor of Hsp90. The functional significance of nucleotide binding by Hsp90 has remained unclear. Here we present evidence for a slow but clearly detectable ATPase activity in purified Hsp90. Based on a new crystal structure of the NH2-terminal domain of human Hsp90 with bound ADP-Mg and on the structural homology of this domain with the ATPase domain of Escherichia coli DNA gyrase, the residues of Hsp90 critical in ATP binding (D93) and ATP hydrolysis (E47) were identified. The corresponding mutations were made in the yeast Hsp90 homologue, Hsp82, and tested for their ability to functionally replace wild-type Hsp82. Our results show that both ATP binding and hydrolysis are required for Hsp82 function in vivo. The mutant Hsp90 proteins tested are defective in the binding and ATP hydrolysis-dependent cycling of the co-chaperone p23, which is thought to regulate the binding and release of substrate polypeptide from Hsp90. Remarkably, the complete Hsp90 protein is required for ATPase activity and for the interaction with p23, suggesting an intricate allosteric communication between the domains of the Hsp90 dimer. Our results establish Hsp90 as an ATP-dependent chaperone.


MEG in the macaque monkey and human: distinguishing cortical fields in space and time.
Magnetoencephalography (MEG) is an increasingly popular non-invasive tool used to record, on a millisecond timescale, the magnetic field changes generated by cortical neural activity. MEG has the advantage, over fMRI for example, that it is a direct measure of neural activity. In the current investigation we used MEG to measure cortical responses to tactile and auditory stimuli in the macaque monkey. We had two aims. First, we sought to determine whether MEG, a technique that may have low spatial accuracy, could be used to distinguish the location and organization of sensory cortical fields in macaque monkeys, a species with a relatively small brain compared to that of the human. Second, we wanted to examine the temporal dynamics of cortical responses in the macaque monkey relative to the human. We recorded MEG data from anesthetized monkeys and, for comparison, from awake humans that were presented with simple tactile and auditory stimuli. Neural source reconstruction of MEG data showed that primary somatosensory and auditory cortex could be differentiated and, further, that separate representations of the digit and lip within somatosensory cortex could be identified in macaque monkeys as well as humans. We compared the latencies of activity from monkey and human data for the three stimulation types and proposed a correspondence between the neural responses of the two species. We thus demonstrate the feasibility of using MEG in the macaque monkey and provide a non-human primate model for examining the relationship between external evoked magnetic fields and their underlying neural sources.


Multi-Compartment, Early Disruption of cGMP and cAMP Signalling in Cardiac Myocytes from the mdx Model of Duchenne Muscular Dystrophy.
Duchenne muscular dystrophy (DMD) is the most frequent and severe form of muscular dystrophy. The disease presents with progressive body-wide muscle deterioration and, with recent advances in respiratory care, cardiac involvement is an important cause of morbidity and mortality. DMD is caused by mutations in the dystrophin gene resulting in the absence of dystrophin and, consequently, disturbance of other proteins that form the dystrophin-associated protein complex (DAPC), including neuronal nitric oxide synthase (nNOS). The molecular mechanisms that link the absence of dystrophin with the alteration of cardiac function remain poorly understood but disruption of NO-cGMP signalling, mishandling of calcium and mitochondrial disturbances have been hypothesized to play a role. cGMP and cAMP are second messengers that are key in the regulation of cardiac myocyte function and disruption of cyclic nucleotide signalling leads to cardiomyopathy. cGMP and cAMP signals are compartmentalised and local regulation relies on the activity of phosphodiesterases (PDEs). Here, using genetically encoded FRET reporters targeted to distinct subcellular compartments of neonatal cardiac myocytes from the DMD mouse model mdx, we investigate whether lack of dystrophin disrupts local cyclic nucleotide signalling, thus potentially providing an early trigger for the development of cardiomyopathy. Our data show a significant alteration of both basal and stimulated cyclic nucleotide levels in all compartments investigated, as well as a complex reorganization of local PDE activities.


Targeted Metabolomic Analysis of Serum Fatty Acids for the Prediction of Autoimmune Diseases.
Autoimmune diseases (ADs) are rapidly increasing worldwide and accumulating data support a key role of disrupted metabolism in ADs. This study aimed to identify an improved combination of Total Fatty Acids (TFAs) biomarkers as a predictive factor for the presence of autoimmune diseases. A retrospective nested case-control study was conducted in 403 individuals. In the case group, 240 patients diagnosed with rheumatoid arthritis, thyroid disease, multiple sclerosis, vitiligo, psoriasis, inflammatory bowel disease, and other AD were included and compared to 163 healthy individuals. Targeted metabolomic analysis of serum TFAs was performed using GC-MS, and 28 variables were used as input for the predictive models. The primary analysis identified 12 variables that were statistically significantly different between the two groups, and metabolite-metabolite correlation analysis revealed 653 significant correlation coefficients with 90% level of significance (p < 0.05). Three predictive models were developed, namely (a) a logistic regression based on Principal Component Analysis (PCA), (b) a straightforward logistic regression model and (c) an Artificial Neural Network (ANN) model. PCA and straightforward logistic regression analysis, indicated reasonably well adequacy (74.7 and 78.9%, respectively). For the ANN, a model using two hidden layers and 11 variables was developed, resulting in 76.2% total predictive accuracy. The models identified important biomarkers: lauric acid (C12:0), myristic acid (C14:0), stearic acid (C18:0), lignoceric acid (C24:0), palmitic acid (C16:0) and heptadecanoic acid (C17:0) among saturated fatty acids, Cis-10-pentadecanoic acid (C15:1), Cis-11-eicosenoic acid (C20:1n9), and erucic acid (C22:1n9) among monounsaturated fatty acids and the Gamma-linolenic acid (C18:3n6) polyunsaturated fatty acid. The metabolic pathways of the candidate biomarkers are discussed in relation to ADs. The findings indicate that the metabolic profile of serum TFAs is associated with the presence of ADs and can be an adjunct tool for the early diagnosis of ADs.


Long-term outcomes of clinical transplantation of pancreatic islets with uncontrolled donors after cardiac death: a multicenter experience in Japan.
Pancreatic islet transplantation has emerged as an effective treatment for type 1 diabetes mellitus, but its use is limited due to an insufficient supply of cadaveric pancreata. In Japan, uncontrolled donors after cardiac death (DCD) are not deemed to be suitable for whole-organ pancreatic transplantation, and can provide a source of pancreas for islet transplantation. However, the long-term outcomes and utility of uncontrolled DCD in the clinical setting remain controversial. Here, we summarize the long-term outcomes of islet transplantation employing uncontrolled DCD as reported to the Japan Islet Transplantation Registry. Sixty-four isolations and 34 transplantations of pancreatic islets were conducted in 18 subjects with type 1 diabetes mellitus under the cover of immunosuppression with basiliximab, sirolimus, and tacrolimus. All donors were uncontrolled DCD at the time of harvesting. The mean follow-up time was 76 months. Of the 18 recipients, 8, 4, and 6 recipients received 1, 2, and 3 islet infusions, respectively. Overall graft survivals (defined as a C-peptide level ≥0.3 ng/mL) were 72.2%, 44.4%, and 22.2% at 1, 2, and 5 years, respectively, whereas the corresponding graft survivals after multiple infusions were 90.0%, 70.0%, and 30.0%, respectively. Three of these recipients achieved insulin independence in 14, 79, and 215 days. HbA1c levels and the requirement of exogenous insulin were improved before loss of graft function. All recipients became free of severe hypoglycemia unawareness, however, at least 5 of 14 patients who had graft failure experienced recurrence of severe hypoglycemia after the loss of graft function. Islet transplantation from DCD can relieve glucose instability and problems with hypoglycemia when the graft is functioning. However, islets from uncontrolled DCD may be associated with reduced long-term graft survival. Further improvements in the clinical outcome by modification of islet isolation/transplantation protocols are necessary to establish islet transplantation using DCD.


Do general practitioners "miss" depression in elderly patients?
In a study of the prevalence of depression in 235 elderly patients who attended general practice surgeries less than 12% of the disagreement between the research assessment of depression and the general practitioner's assessment was due to "missed" depression. There were, however, low rates of referral and of treatment with antidepressant drugs. If these findings are confirmed the study of the management and outcome of depression in such patients may be more rewarding than attempts to improve the recognition of depression.


Object-Based Point Cloud Analysis of Full-Waveform Airborne Laser Scanning Data for Urban Vegetation Classification.
Airborne laser scanning (ALS) is a remote sensing technique well-suited for 3D vegetation mapping and structure characterization because the emitted laser pulses are able to penetrate small gaps in the vegetation canopy. The backscattered echoes from the foliage, woody vegetation, the terrain, and other objects are detected, leading to a cloud of points. Higher echo densities (> 20 echoes/m2) and additional classification variables from full-waveform (FWF) ALS data, namely echo amplitude, echo width and information on multiple echoes from one shot, offer new possibilities in classifying the ALS point cloud. Currently FWF sensor information is hardly used for classification purposes. This contribution presents an object-based point cloud analysis (OBPA) approach, combining segmentation and classification of the 3D FWF ALS points designed to detect tall vegetation in urban environments. The definition tall vegetation includes trees and shrubs, but excludes grassland and herbage. In the applied procedure FWF ALS echoes are segmented by a seeded region growing procedure. All echoes sorted descending by their surface roughness are used as seed points. Segments are grown based on echo width homogeneity. Next, segment statistics (mean, standard deviation, and coefficient of variation) are calculated by aggregating echo features such as amplitude and surface roughness. For classification a rule base is derived automatically from a training area using a statistical classification tree. To demonstrate our method we present data of three sites with around 500,000 echoes each. The accuracy of the classified vegetation segments is evaluated for two independent validation sites. In a point-wise error assessment, where the classification is compared with manually classified 3D points, completeness and correctness better than 90% are reached for the validation sites. In comparison to many other algorithms the proposed 3D point classification works on the original measurements directly, i.e. the acquired points. Gridding of the data is not necessary, a process which is inherently coupled to loss of data and precision. The 3D properties provide especially a good separability of buildings and terrain points respectively, if they are occluded by vegetation.


Efficacy of aqueous extract of Hippophae rhamnoides and its bio-active flavonoids against hypoxia-induced cell death.
To investigate the protective efficacy of aqueous extract of Hippophae rhamnoides against chronic hypoxic injury using primary rat hepatocytes. The extract was prepared using maceration method and characterized by its phenolic and flavonoid content and chemical antioxidant capacity using ferric reducing antioxidant power assay. Hepatocytes were maintained in hypoxia chamber (3% and 1% oxygen) for 72 h. The cells kept under normoxic condition served as control. The cells were treated with the extract and flavonoids; isorhamentin, kaempferol or qurecetin-3-galactoside. After the end of exposure period; cell survival, reactive oxygen species (ROS), leakage of lactate dehydrogenase (LDH), alanine aminotransferase (ALT), aspartate aminotransferase (AST), reduced glutathione (GSH), glutathione peroxidase (GPx), and superoxide dismutase (SOD) levels were measured. The extract showed presence of high phenolic and flavonoid content with significant antioxidant activity in chemical assay. The cell exposed to hypoxia showed concentration dependent cell death and harbored higher reactive oxygen species. In addition, these cells showed significant leakage of intracellular LDH, ALT, and AST accompanied by the diminished levels/activities of GSH, GPx, and SOD. The treatment of cells with aqueous extract of H. rhamnoides reduced hypoxia-induced cell death and prevented increase in ROS levels and leakage of intracellular LDH, ALT, and AST from cells. Moreover, these cells maintained better levels/activities of GSH, GPx, and SOD in comparison to the respective controls. The major flavonoids present in aqueous extract of H. rhamnoides; quercetin-3-galactoside, kaempferol, and isorhamentin also prevented hypoxia induced cell injury individually or in combination, however, the protection offered by these compounds taken together could not match to that of the extract. Overall the findings reveal significance of aqueous extract of H. rhamnoides in controlling ROS-meditated hypoxic injury in cells and can be useful in many hepatic complications.


Do adolescents understand the items of the European Health Literacy Survey Questionnaire (HLS-EU-Q47) - German version? Findings from cognitive interviews of the project "Measurement of Health Literacy Among Adolescents" (MOHLAA) in Germany.
In Germany, there are no measurement tools to assess the general health literacy of adolescents. The aim of the study "Measurement of Health Literacy Among Adolescents" (MOHLAA) is to develop such a tool for use among adolescents aged 14-17. The German version of the European Health Literacy Survey Questionnaire (HLS-EU-Q47-GER) served as a blueprint for the development of the tool. The present study examined the extent to which the HLS-EU-Q47-GER can be applied to the measurement of general health literacy in adolescents. The applicability of the HLS-EU-Q47-GER for adolescents was tested qualitatively using cognitive interviewing (CI). Purposive sampling was used to achieve an equal distribution of participants regarding age groups, educational backgrounds and gender. CI was standardized on the basis of an interview guide. Verbal probing and the retrospective think-aloud technique were applied. The interviews were audio-recorded, transcribed and analyzed using the criteria of theory-based analysis, which were derived from the model of cognitive processes. The analysis focused on identifying terms and questions that were difficult to understand and on scrutinizing the extent to which the content of the items is appropriate for assessing adolescents' health literacy. Adolescent respondents were unfamiliar with some terms of the HLS-EU-Q47-GER or provided heterogeneous interpretations of the terms. They had limited or no experience regarding some health-related tasks in health care and disease prevention that are addressed by HLS-EU-Q-items. A few items seemed to be too "difficult" to answer due to a high abstraction level or because they lacked any reference to the everyday lives of youth. Despite comprehension problems with some of the HLS-EU items, the respondents assessed the covered health-related tasks as "very easy" or "fairly easy". CI stressed the importance of interpersonal agents, especially parents, in helping adolescents understand and judge the reliability of health information. The results of CI indicated that the applicability of the HLS-EU-Q47-GER to the measurement of general health literacy among adolescents aged 14-17 is limited. In order to prevent biased data, some items of the questionnaire should be adjusted to adolescents' state of development and experiences with health care and disease prevention.


Borrelia burgdorferi infection and immunity in mice deficient in the fifth component of complement.
When immunocompetent mice are inoculated with Borrelia burgdorferi, they develop acute arthritis and carditis that undergo spontaneous regression despite the persistence of infection. Specific T- and/or B-cell immunity appears to be necessary for resolution of disease manifestations. Humoral immune responses to B. burgdorferi are also important in prevention of B. burgdorferi infection, in that passive transfer of immune sera or protective monoclonal antibodies prevents the spirochete from establishing infection. It has previously been suggested that complement is necessary for effective antibody-mediated host responses against B. burgdorferi. To investigate the role of complement in the pathogenesis and prevention of Lyme disease, we compared the responses to B. burgdorferi challenge inoculation of mice genetically deficient in the fifth component of complement (C5) with those of C5-sufficient mice. All C5-deficient strains tested were susceptible to B. burgdorferi infection, and disease manifestations underwent regression in a similar time-course to those of complement-sufficient mice. Moreover, passive immunization of C5-deficient mice with either immune rabbit sera or neutralizing monoclonal antibody protected them from challenge infection. These results demonstrate that the expression of Lyme disease is not altered in mice deficient in C5 and that C5-mediated complement activation is not necessary for antibody-mediated protection from infection.


Strong Induction of Minor Terpenes in Italian Cypress, Cupressus sempervirens, in Response to Infection by the Fungus Seiridium cardinale.
Seiridium cardinale, the main fungal pathogen responsible for cypress bark canker, is the largest threat to cypresses worldwide. The terpene response of canker-resistant clones of Italian cypress, Cupressus sempervirens, to two differently aggressive isolates of S. cardinale was studied. Phloem terpene concentrations, foliar terpene concentrations, as well as foliar terpene emission rates were analyzed 1, 10, 30, and 90 days after artificial inoculation with fungal isolates. The phloem surrounding the inoculation point exhibited de novo production of four oxygenated monoterpenes and two unidentified terpenes. The concentrations of several constitutive mono- and diterpenes increased strongly (especially α-thujene, sabinene, terpinolene, terpinen-4-ol, oxygenated monoterpenes, manool, and two unidentified diterpenes) as the infection progressed. The proportion of minor terpenes in the infected cypresses increased markedly from the first day after inoculation (from 10% in the control to 30-50% in the infected treatments). Foliar concentrations showed no clear trend, but emission rates peaked at day 10 in infected trees, with higher δ-3-carene (15-fold) and total monoterpene (10-fold) emissions than the control. No substantial differences were found among cypresses infected by the two fungal isolates. These results suggest that cypresses activate several direct and indirect chemical defense mechanisms after infection by S. cardinale.


Influence of maternal obesity on the long-term health of offspring.
In addition to immediate implications for pregnancy complications, increasing evidence implicates maternal obesity as a major determinant of offspring health during childhood and later adult life. Observational studies provide evidence for effects of maternal obesity on her offspring's risks of obesity, coronary heart disease, stroke, type 2 diabetes, and asthma. Maternal obesity could also lead to poorer cognitive performance and increased risk of neurodevelopmental disorders, including cerebral palsy. Preliminary evidence suggests potential implications for immune and infectious-disease-related outcomes. Insights from experimental studies support causal effects of maternal obesity on offspring outcomes, which are mediated at least partly through changes in epigenetic processes, such as alterations in DNA methylation, and perhaps through alterations in the gut microbiome. Although the offspring of obese women who lose weight before pregnancy have a reduced risk of obesity, few controlled intervention studies have been done in which maternal obesity is reversed and the consequences for offspring have been examined. Because the long-term effects of maternal obesity could have profound public health implications, there is an urgent need for studies on causality, underlying mechanisms, and effective interventions to reverse the epidemic of obesity in women of childbearing age and to mitigate consequences for offspring.


Energy expenditure and body composition changes after an isocaloric ketogenic diet in overweight and obese men.
The carbohydrate-insulin model of obesity posits that habitual consumption of a high-carbohydrate diet sequesters fat within adipose tissue because of hyperinsulinemia and results in adaptive suppression of energy expenditure (EE). Therefore, isocaloric exchange of dietary carbohydrate for fat is predicted to result in increased EE, increased fat oxidation, and loss of body fat. In contrast, a more conventional view that "a calorie is a calorie" predicts that isocaloric variations in dietary carbohydrate and fat will have no physiologically important effects on EE or body fat. We investigated whether an isocaloric low-carbohydrate ketogenic diet (KD) is associated with changes in EE, respiratory quotient (RQ), and body composition. Seventeen overweight or obese men were admitted to metabolic wards, where they consumed a high-carbohydrate baseline diet (BD) for 4 wk followed by 4 wk of an isocaloric KD with clamped protein. Subjects spent 2 consecutive days each week residing in metabolic chambers to measure changes in EE (EEchamber), sleeping EE (SEE), and RQ. Body composition changes were measured by dual-energy X-ray absorptiometry. Average EE during the final 2 wk of the BD and KD periods was measured by doubly labeled water (EEDLW). Subjects lost weight and body fat throughout the study corresponding to an overall negative energy balance of ∼300 kcal/d. Compared with BD, the KD coincided with increased EEchamber (57 ± 13 kcal/d, P = 0.0004) and SEE (89 ± 14 kcal/d, P < 0.0001) and decreased RQ (-0.111 ± 0.003, P < 0.0001). EEDLW increased by 151 ± 63 kcal/d (P = 0.03). Body fat loss slowed during the KD and coincided with increased protein utilization and loss of fat-free mass. The isocaloric KD was not accompanied by increased body fat loss but was associated with relatively small increases in EE that were near the limits of detection with the use of state-of-the-art technology. This trial was registered at clinicaltrials.gov as NCT01967563.


Successive light-induced two electron transfers in a Ru-Fe supramolecular assembly: from Ru-Fe(ii)-OH2 to Ru-Fe(iv)-oxo.
In the present work we describe the synthesis and study of a RuII-FeII chromophore-catalyst assembly designed to perform the light-induced activation of an iron bound water molecule and subsequent photo-driven oxidation of a substrate. Using a series of spectroscopic techniques, we demonstrate that excitation of the chromophore unit with 450 nm light, in the presence of a sacrificial electron acceptor, triggers a cascade of electron transfers leading to the formation of a high valent iron(iv)-oxo center from an iron(ii)-bound water molecule. The activity of this catalytic center is illustrated by the oxidation of triphenyl phosphine.


A DNA segment encoding two genes very tightly linked to Huntington's disease.
The discovery of D4S10, an anonymous DNA marker genetically linked to Huntington's disease (HD), introduced the capacity for limited presymptomatic diagnosis in this late-onset neurodegenerative disorder and raised the hope of cloning and characterizing the defect based on its chromosomal location. Progress on both fronts has been limited by the absence of additional DNA markers closer to the HD gene. An anonymous DNA locus, D4S43, has now been found that shows extremely tight linkage to HD. Like the disease gene, D4S43 is located in the most distal region of the chromosome 4 short arm, flanked by D4S10 and the telomere. In three extended HD kindreds, D4S43 displays no recombination with HD, placing it within 0 to 1.5 centimorgans of the genetic defect. Expansion of the D4S43 region to include 108 kilobases of cloned DNA has allowed identification of eight restriction fragment length polymorphisms and at least two independent coding segments. In the absence of crossovers, these genes must be considered candidates for the site of the HD defect, although the D4S43 restriction fragment length polymorphisms do not display linkage disequilibrium with the disease gene.


Reduction of severe acute maternal morbidity and maternal mortality in Thyolo District, Malawi: the impact of obstetric audit.
Critical incident audit and feedback are recommended interventions to improve the quality of obstetric care. To evaluate the effect of audit at district level in Thyolo, Malawi, we assessed the incidence of facility-based severe maternal complications (severe acute maternal morbidity (SAMM) and maternal mortality) during two years of audit and feedback. Between September 2007 and September 2009, we included all cases of maternal mortality and SAMM that occurred in Thyolo District Hospital, the main referral facility in the area, using validated disease-specific criteria. During two- to three-weekly audit sessions, health workers and managers identified substandard care factors. Resulting recommendations were implemented and followed up. Feedback was given during subsequent sessions. A linear regression analysis was performed on facility-based severe maternal complications. During the two-year study period, 386 women were included: 46 died and 340 sustained SAMM, giving a case fatality rate of 11.9%. Forty-five cases out of the 386 inclusions were audited in plenary with hospital staff. There was a reduction of 3.1 women with severe maternal complications per 1000 deliveries in the district health facilities, from 13.5 per 1000 deliveries in the beginning to 10.4 per 1000 deliveries at the end of the study period. The incidence of uterine rupture and major obstetric hemorrhage reduced considerably (from 3.5 to 0.2 and from 5.9 to 2.6 per 1000 facility deliveries respectively). Our findings indicate that audit and feedback have the potential to reduce serious maternal complications including maternal mortality. Complications like major hemorrhage and uterine rupture that require relatively straightforward intrapartum emergency management are easier to reduce than those which require uptake of improved antenatal care (eclampsia) or timely intravenous medication or HIV-treatment (peripartum infections).


Effects of gestational exposure to di-n-butyl phthalate and mineral oil on testis development of the Mongolian gerbil.
Phthalate esters are endocrine disrupters that can affect the development of the testis in a species-specific manner. However, their interference in the male gonads of the Mongolian gerbil is unknown. The aim of the present study was to evaluate whether gestational exposure to di-n-butyl phthalate (DBP) interferes with the development of the gerbil testis during the first six weeks of life. Males were evaluated at 1, 7, 14, 28, 35 and 42 days of age in an untreated (control) group or groups exposed from 8 to 23 days gestation to DBP (100mgkg-1day-1 in mineral oil) or vehicle by maternal gavage. DBP exposure impaired cell proliferation within the seminiferous cords at birth, but increased proliferation at the end of the first week, when higher testosterone concentrations were observed. The vehicle (mineral oil) reduced the total number of gonocytes and attenuated the decrease in testosterone concentrations at 7 days. The vehicle also altered gonocyte relocation at 14 days and increased oestrogen concentrations at 28 days by approximately 112%. In summary, both DBP and oil interfered in gonadal development and testosterone plasma concentrations in the first week of postnatal life. However, the changes observed at the beginning of puberty were not seen after exposure to DBP, indicating a more harmful effect of mineral oil in this period.


Differential regulation of prostaglandin F(2alpha) receptor isoforms by protein kinase C.
Prostaglandin F(2alpha) receptors (FP) are G protein-coupled receptors that bind prostaglandin F(2alpha) (PGF(2alpha)), resulting in the activation of an inositol phosphate (IP) second messenger pathway. Alternative mRNA splicing generates two FP receptor isoforms. These isoforms, designated FP(A) and FP(B), are otherwise identical except for their carboxyl termini. FP(B) is essentially a truncated version of FP(A) that lacks the 46 carboxyl-terminal amino acids, including four putative protein kinase C (PKC) phosphorylation sites. Until now, functional differences between these FP receptor isoforms have not been identified. We now report that pretreatment with the PKC inhibitor bisindolylmaleimide I enhanced PGF(2alpha)-stimulated IP accumulation in transfected cells stably expressing the FP(A) isoform but not in cells stably expressing the FP(B) isoform. Whole-cell phosphorylation experiments showed a strong agonist-dependent phosphorylation of the FP(A) isoform but little or no phosphorylation of the FP(B). Pretreatment of cells with bisindolylmaleimide I decreased PGF(2alpha)-stimulated phosphorylation of the FP(A) isoform consistent with a PKC-dependent phosphorylation. In vitro phosphorylation of an FP(A) carboxyl-terminal fusion protein by recombinant PKCalpha showed that the carboxyl terminus of the FP(A) is a substrate for PKC. These results suggest that PKC-dependent phosphorylation is responsible for differential regulation of second messenger signaling by FP prostanoid receptor isoforms.


Enhancement of Friction by Water Intercalated between Graphene and Mica.
Common experience shows that friction converts mechanical energy into heat. The first part of this process is vibrational excitation of atoms at the interface between rubbing bodies. The second part is the removal of the vibration energy by transferring it from the interface to the substrate. However, it is difficult to disentangle the excitation and energy transfer processes. We solved this by using a system consisting of a SiO2-terminated tip sliding over graphene deposited on mica with intercalated water between them. The intercalated water was found to increase friction by a factor of ∼3 relative to dry mica. Density functional theory calculations show that water broadens the spectral range of graphene vibrations-particularly the low-frequency flexural modes-thus providing new excitation channels and also by increasing the overlap with the atomic vibrations of the mica substrate, which facilitates coupling and energy transfer.


Patient engagement with surgical site infection prevention: an expert panel perspective.
Despite remarkable developments in the use of surgical techniques, ergonomic advancements in the operating room, and implementation of bundles, surgical site infections (SSIs) remain a substantial burden, associated with increased morbidity, mortality and healthcare costs. National and international recommendations to prevent SSIs have been published, including recent guidelines by the World Health Organization, but implementation into clinical practice remains an unresolved issue. SSI improvement programs require an integrative approach with measures taken during the pre-, intra- and postoperative care from the numerous stakeholders involved. The current SSI prevention strategies have focused mainly on the role of healthcare workers (HCWs) and procedure related risk factors. The importance and influence of patient participation is becoming an increasingly important concept and advocated as a means to improve patient safety. Novel interventions supporting an active participative role within SSI prevention programs have not been assessed. Empowering patients with information they require to engage in the process of SSI prevention could play a major role for the implementation of recommendations. Based on available scientific evidence, a panel of experts evaluated options for patient involvement in order to provide pragmatic recommendations for pre-, intra- and postoperative activities for the prevention of SSIs. Recommendations were based on existing guidelines and expert opinion. As a result, 9 recommendations for the surgical patient are presented here, including a practice brief in the form of a patient information leaflet. HCWs can use this information to educate patients and allow patient engagement.


Cross-linking amine-rich compounds into high performing selective CO2 absorbents.
Amine-based absorbents play a central role in CO2 sequestration and utilization. Amines react selectively with CO2, but a drawback is the unproductive weight of solvent or support in the absorbent. Efforts have focused on metal organic frameworks (MOFs) reaching extremely high CO2 capacity, but limited selectivity to N2 and CH4, and decreased uptake at higher temperatures. A desirable system would have selectivity (cf. amine) and high capacity (cf. MOF), but also increased adsorption at higher temperatures. Here, we demonstrate a proof-of-concept where polyethyleneimine (PEI) is converted to a high capacity and highly selective CO2 absorbent using buckminsterfullerene (C(60)) as a cross-linker. PEI-C(60) (CO2 absorption of 0.14 g/g at 0.1 bar/90 °C) is compared to one of the best MOFs, Mg-MOF-74 (0.06 g/g at 0.1 bar/90 °C), and does not absorb any measurable amount of CH4 at 50 bar. Thus, PEI-C(60) can perform better than MOFs in the sweetening of natural gas.


A survey of owners' perceptions and experiences of radioiodine treatment of feline hyperthyroidism in the UK.
The efficacy of radioiodine treatment of feline hyperthyroidism is well established; however, limited information is known about owners' perceptions or experiences of radioiodine. This study aimed to examine factors that influence owner treatment choices and their opinions following radioiodine. Surveys were sent to owners of cats referred for radioiodine treatment between 2002 and 2011 (radioiodine group; 264 cats) and owners of non-radioiodine-treated hyperthyroid cats seen at first-opinion practices (control group; 199 cats). The response rate was 67.0% (310 returned: 175 radioiodine, 135 control). Of 135 controls, 72 (53.3%) were unaware of radioiodine as a treatment option. Owners of cats ⩾15 years old and uninsured cats were less likely to pursue radioiodine. Cost of treatment, travel distance, potential human or animal health risks and waiting periods for radioiodine had a low impact on owners' treatment choice. Owners reported a moderate level of concern about treatment hospitalisation length, which included (158 respondents) the possibility of the cat being unhappy 130 (82.3%), owner missing the cat 102 (64.6%), inappetence 50 (31.6%), other pets missing the cat 32 (20.3%), development of co-morbid disease 28 (17.7%) and side effects 25 (15.8%). Owners assessed their cat's quality of life on a scale of 1 (very poor) to 10 (excellent), as 4 (4) (median [interquartile range]) pre-radioiodine (134 respondents) and 9 (2) post-radioiodine (131 respondents). Of 132 respondents, 121 (91.7%) were happy with their decision to choose radioiodine. The results of this questionnaire may assist veterinarians in addressing common owner concerns when discussing radioiodine as a treatment option for hyperthyroidism.


Spin Relays Enable Efficient Long-Range Heteronuclear Signal Amplification By Reversible Exchange.
A systematic experimental study is reported on the polarization transfer to distant spins, which do not directly bind to the polarization transfer complexes employed in Signal Amplification By Reversible Exchange (SABRE) experiments. Both, long-range transfer to protons and long-range transfer to heteronuclei i.e. 13C and 15N are examined. Selective destruction of hyperpolarization on 1H, 13C, and 15N sites is employed, followed by their re-hyperpolarization from neighboring spins within the molecules of interest (pyridine for 1H studies and metronidazole-15N2-13C2 for 13C and 15N studies). We conclude that long-range sites can be efficiently hyperpolarized when a network of spin-½ nuclei enables relayed polarization transfer (i.e. via short-range interactions between sites). In case of proton SABRE in the milli-Tesla regime, a relay network consisting of protons only is sufficient. However, in case 13C and 15N are targeted (i.e. via SABRE in SHield Enables Alignment Transfer to Heteronuclei or SABRE-SHEATH experiment), the presence of a heteronuclear network (e.g. consisting of 15N) enables a relay mechanism that is significantly more efficient than the direct transfer of spin order from para-H2-derived hydrides.


Differential effects of olanzapine and clozapine on plasma levels of adipocytokines and total ghrelin.
Second-generation antipsychotics (SGAs) have been associated with an increased liability for weight gain and metabolic side effects. Among SGAs, clozapine and olanzapine had great liability to induce weight gain and metabolic adverse reactions. Leptin, adiponectin, and total ghrelin play important roles in energy homeostasis and are suggested to be biomarkers of metabolic disturbances. The purpose of the present study was to investigate the differential effects of antipsychotics (olanzapine and clozapine) on the levels of adipocytokines (leptin and adiponectin) and total ghrelin. Three hundred and thirty-three patients with schizophrenia under clozapine or olanzapine monotherapy were recruited. Control participants were recruited from a healthy community population based on a health investigation (N=119). Fasting blood samples for glucose, cholesterol, triglycerides, leptin, adiponectin, and total ghrelin were analyzed. There were significant differences in the levels of cholesterol, triglycerides, and glucose between these three groups. Post hoc comparisons showed that the olanzapine group had the highest levels of cholesterol and triglycerides. The levels of leptin, adiponectin, and total ghrelin were also significantly different between the three groups after controlling age and body mass index (BMI). Post hoc comparisons showed that the olanzapine group had the lowest levels of adiponectin and total ghrelin. The present study found that the uses of olanzapine and clozapine were associated with changes in adipocytokines and total ghrelin, even after adjusting potential confounding factors. Olanzapine had greater influences on adiponectin and total ghrelin than clozapine. The changes in adipocytokines and total ghrelin were a direct effect of antipsychotics on hormonal pathways of energy homeostasis, rather than the result of weight gain.


Safety and Efficacy of Nivolumab in Brain Metastases From Renal Cell Carcinoma: Results of the GETUG-AFU 26 NIVOREN Multicenter Phase II Study.
Nivolumab is standard of care for patients with metastatic clear cell renal cell carcinoma (ccRCC) after failure of antiangiogenic therapies, but its activity on brain metastases from ccRCC remains unknown, because these patients were excluded from pivotal studies. We aimed to assess the activity of nivolumab in this population. The GETUG-AFU 26 NIVOREN phase II trial assessed the activity and safety of nivolumab in patients with metastatic ccRCC who failed vascular endothelial growth factor-directed therapies (ClinicalTrials.gov identifier: NCT03013335). Patients with asymptomatic brain metastases were prospectively identified and underwent dedicated brain evaluation. Two cohorts were constituted: cohort A comprised patients with previously untreated brain metastases, and cohort B comprised patients whose brain metastases underwent prior therapy. The primary end point was intracranial response rate in cohort A. Seventy-three patients with brain metastases were included: 39 in cohort A and 34 in cohort B. Intracranial response rate was 12% in cohort A; no objective response was reported in patients with brain lesions that were multiple or larger than 1 cm. Median intracranial progression-free survival was 2.7 months (95% CI, 2.3 to 4.6 months) in cohort A and 4.8 months (95% CI, 3.0 to 8.0 months) in cohort B, with adjusted hazard ratio of 2.04 (95% CI, 1.08 to 3.83). Overall survival rate at 12 months was 67% (95% CI, 49.6% to 79.1%) in cohort A and 59% (95% CI, 40.6% to 73.2%) in cohort B. Most patients in cohort A (72%) needed subsequent focal brain therapy. Nivolumab was well tolerated, with no unexpected toxicity. Nivolumab activity is limited in patients with untreated brain metastases from ccRCC. Brain imaging and focal therapy should be considered before immune checkpoint inhibitors in patients with metastatic ccRCC.


A Cross-Species Analysis of Animal Models for the Investigation of Preterm Birth Mechanisms.
Spontaneous preterm birth is the leading cause of neonatal morbidity and mortality worldwide. The ability to examine the exact mechanisms underlying this syndrome in humans is limited. Therefore, the study of animal models is critical to unraveling the key physiologic mechanisms that control the timing of birth. The purpose of this review is to facilitate enhanced assimilation of the literature on animal models of preterm birth by a broad range of investigators. Using classical systematic and informatics search techniques of the available literature through 2012, a database of intact animal models was generated. Research librarians generated a list of articles using multiple databases. From these articles, a comprehensive list of Medical Subject Headings (MeSH) was created. Using mathematical modeling, significant MeSH descriptors were determined, and a MEDLINE search algorithm was created. The articles were reviewed for mechanism of labor induction categorized by species. Existing animal models of preterm birth comprise specific interventions to induce preterm birth, as no animal model was identified that exhibits natural spontaneous preterm birth at an incidence comparable to that of the humans. A search algorithm was developed which when used results in a comprehensive list of agents used to induce preterm delivery in a host of animal species. The evolution of 3 specific animal models--sheep, mice, and rats--has demonstrated a clear shift in focus in the literature from endocrine to inflammatory agents of preterm birth induction. The process of developing a search algorithm to provide efficient access to information on animal models of preterm birth illustrates the need for a more precise organization of the literature to allow the investigator to focus on distinctly maternal versus fetal outcomes.


Anterior cingulate cortex, error detection, and the online monitoring of performance.
An unresolved question in neuroscience and psychology is how the brain monitors performance to regulate behavior. It has been proposed that the anterior cingulate cortex (ACC), on the medial surface of the frontal lobe, contributes to performance monitoring by detecting errors. In this study, event-related functional magnetic resonance imaging was used to examine ACC function. Results confirm that this region shows activity during erroneous responses. However, activity was also observed in the same region during correct responses under conditions of increased response competition. This suggests that the ACC detects conditions under which errors are likely to occur rather than errors themselves.


Agarose-Based Hydrogels as Suitable Bioprinting Materials for Tissue Engineering.
Hydrogels are useful materials as scaffolds for tissue engineering applications. Using hydrogels with additive manufacturing techniques has typically required the addition of techniques such as cross-linking or printing in sacrificial materials that negatively impact tissue growth to remedy inconsistencies in print fidelity. Thus, there is a need for bioinks that can directly print cell-laden constructs. In this study, agarose-based hydrogels commonly used for cartilage tissue engineering were compared to Pluronic, a hydrogel with established printing capabilities. Moreover, new material mixtures were developed for bioprinting by combining alginate and agarose. We compared mechanical and rheological properties, including yield stress, storage modulus, and shear thinning, as well as construct shape fidelity to assess their potential as a bioink for cell-based tissue engineering. The rheological properties and printability of agarose-alginate gels were statistically similar to those of Pluronic for all tests (p > 0.05). Alginate-agarose composites prepared with 5% w/v (3:2 agarose to alginate ratio) demonstrated excellent cell viability over a 28-day culture period (>∼70% cell survival at day 28) as well matrix production over the same period. Therefore, agarose-alginate mixtures showed the greatest potential as an effective bioink for additive manufacturing of biological materials for cartilage tissue engineering.


Ethical questions regarding health insurance in India.
Improved health and healthcare are of vital concern to the welfare of Indian society. The nascent health insurance system of the country is experiencing an explosive expansion and various models of health insurance provision are under trial by different agencies. Since the country has been relatively late in introducing health insurance, it can study the effects of different systems of healthcare and insurance and develop a system of health coverage which addresses the unique social character of our country as well as the ethical questions of comprehensiveness and inclusion. This article seeks to explore these issues in detail.


Stability and effect on the soft tissue profile of mandibular setback with sagittal split osteotomy and rigid internal fixation.
The stability and the effect on the soft tissue profile of mandibular setback with sagittal split osteotomy and rigid internal fixation was studied prospectively in 29 consecutive patients, 17 to 55 years old, who had mesial occlusion. Profile cephalometric radiographs were taken immediately before and short after surgery as well as 14 months postsurgically. The mean setback of the mandible 1 to 8 days after surgery was 6.0 mm. On average, there was also a small decrease in the anterior facial height. The setback relapsed an average of 1.1 to 1.3 mm, so that at the follow-up the mean net effect was 80%. In eight patients, however, the mandible had retruded slightly further. During the follow-up period there was a further small decrease in the anterior facial height. The net effects on the labial fold and the soft tissue chin were closely correlated with those on their underlying hard structures. On average, the labial fold and the chin followed the hard structures to 106% and 107%, respectively. The mean net effect for the lower lip was only 88% of the setback of the mandibular incisor. The soft tissue facial height followed the decrease in the skeletal facial height to 60%. The setback procedure also affected the upper lip, which, on average, retruded and lengthened 1.0 to 2.0 mm.


Systemic lupus erythematosus in a multiethnic US cohort (LUMINA). XXV. Smoking, older age, disease activity, lupus anticoagulant, and glucocorticoid dose as risk factors for the occurrence of venous thrombosis in lupus patients.
Venous thrombosis is a relatively frequent and serious complication in systemic lupus erythematosus (SLE) that has been associated with the presence of antiphospholipid antibodies (aPL). However, venous thrombotic events can also be seen in patients without aPL, and only a few patients with aPL develop venous thrombosis. This study was carried out to ascertain other factors contributing to the development of venous thrombosis in SLE. Patients with SLE, ages > or = 16 years with < or = 5 years disease duration and of Hispanic, African American, or Caucasian ethnicity, from LUMINA (LUpus in MInorities, NAture versus nurture), a multiethnic, longitudinal study of outcome, were studied. Selected socioeconomic/demographic, clinical, laboratory, and treatment-exposure variables were compared between patients who developed and those who did not develop venous thrombotic events. Significant and clinically relevant variables were then entered into different multivariable models (Cox proportional hazards and unconditional stepwise logistic regression) to identify independent risk factors associated with the primary outcome. In another model, only patients who developed an event after enrollment (time 0) in the cohort were included. Of 570 LUMINA patients, 51 developed at least 1 venous thrombotic event after SLE diagnosis. In univariable analyses, smoking (P = 0.020), shorter disease duration at time 0 (P = 0.017), serum levels of total cholesterol, low-density lipoprotein, and triglycerides (all P < 0.0001), and presence of lupus anticoagulant (LAC) (P = 0.045) were associated with venous thrombotic events. Survival analyses showed a time-dependent significant association of the primary outcome with smoking (P = 0.008) and a borderline significant association with the presence of LAC (P = 0.070). Multivariable models showed an independent association with smoking, age at time 0, disease activity over time, LAC, mean dose of glucocorticoids, and shorter disease duration at time 0. Venous thrombotic events occur early in the course of SLE. Our data confirm the association between LAC and venous thrombotic events. Smoking, shorter disease duration, older age, disease activity over time, and higher mean daily glucocorticoid dose were identified as additional risk factors for the development of this vascular complication. These findings may have implications for the management of patients with SLE.


Cervical screening with primary HPV testing or cytology in a population of women in which those aged 33 years or younger had previously been offered HPV vaccination: Results of the Compass pilot randomised trial.
Using primary human papillomavirus (HPV) testing for cervical screening increases detection of high-grade cervical intraepithelial neoplastic lesions and invasive cancer (cervical intraepithelial neoplasia grade 2+ [CIN2+]) compared to cytology, but no evaluation has been conducted in a population previously offered HPV vaccination. We aimed to assess colposcopy referral and CIN2+ detection rates for HPV-screened versus cytology-screened women in Australia's HPV-vaccinated population (by 2014, resident women ≤33 years had been age-eligible for HPV vaccination, with 3-dose uptake across age cohorts being about 50%-77%). Compass is an open-label randomised trial of 5-yearly HPV screening versus 2.5-yearly liquid-based cytology (LBC) screening. In the first phase, consenting women aged 25-64 years presenting for routine screening at 47 primary practices in Victoria, Australia, provided a cervical sample and were randomised at a central laboratory at a 1:2:2 allocation to (i) image-read LBC screening with HPV triage of low-grade cytology ('LBC screening'), (ii) HPV screening with those HPV16/18 positive referred to colposcopy and with LBC triage for other oncogenic (OHR) types ('HPV+LBC triage'), or (iii) HPV screening with those HPV16/18 positive referred to colposcopy and with dual-stained cytology triage for OHR types ('HPV+DS triage'). A total of 5,006 eligible women were recruited from 29 October 2013 to 7 November 2014 (recruitment rate 58%); of these, 22% were in the group age-eligible for vaccination. Data on 4,995 participants were analysed after 11 withdrawals; 998 were assigned to, and 995 analysed (99.7%) in, the LBC-screened group; 1,996 assigned to and 1,992 analysed (99.8%) in the HPV+LBC triage group; and 2,012 assigned to and 2,008 analysed (99.8%) in the HPV+DS triage group. No serious trial-related adverse events were reported. The main outcomes were colposcopy referral and detected CIN2+ rates at baseline screening, assessed on an intention-to-treat basis after follow-up of the subgroup of triage-negative women in each arm referred to 12 months of surveillance, and after a further 6 months of follow-up for histological outcomes (dataset closed 31 August 2016). Analysis was adjusted for whether women had been age-eligible for HPV vaccination or not. For the LBC-screened group, the overall referral and detected CIN2+ rates were 27/995 (2.7% [95% CI 1.8%-3.9%]) and 1/995 (0.1% [95% CI 0.0%-0.6%]), respectively; for HPV+LBC triage, these were 75/1,992 (3.8% [95% CI 3.0%-4.7%]) and 20/1,992 (1.0% [95% CI 0.6%-1.5%]); and for HPV+DS triage, these were 79/2,008 (3.9% [95% CI 3.1%-4.9%]) and 24/2,008 (1.2% [95% CI 0.8%-1.6%]) (p = 0.09 for difference in referral rate in LBC versus all HPV-screened women; p = 0.003 for difference in CIN2+ detection rate in LBC versus all HPV-screened women, with p = 0.62 between HPV screening groups). Limitations include that the study population involved a relatively low risk group in a previously well-screened and treated population, that individual women's vaccination status was unknown, and that long-term follow-up data on disease detection in screen-negative women are not yet available. In this study, primary HPV screening was associated with significantly increased detection of high-grade precancerous cervical lesions compared to cytology, in a population where high vaccine uptake was reported in women aged 33 years or younger who were offered vaccination. It had been predicted that increased disease detection might be associated with a transient increase in colposcopy referral rates in the first round of HPV screening, possibly dampened by HPV vaccine effect; in this study, although the point estimates for referral rates in women in each HPV-screened group were 41%-44% higher than in cytology-screened women, the difference in referral rate between cytology- and HPV-screened women was not significant. These findings provide initial support for the implementation of primary HPV screening in vaccinated populations. Australian New Zealand Clinical Trials Registry ACTRN12613001207707.


Repeated measures semiparametric regression using targeted maximum likelihood methodology with application to transcription factor activity discovery.
In longitudinal and repeated measures data analysis, often the goal is to determine the effect of a treatment or aspect on a particular outcome (e.g., disease progression). We consider a semiparametric repeated measures regression model, where the parametric component models effect of the variable of interest and any modification by other covariates. The expectation of this parametric component over the other covariates is a measure of variable importance. Here, we present a targeted maximum likelihood estimator of the finite dimensional regression parameter, which is easily estimated using standard software for generalized estimating equations. The targeted maximum likelihood method provides double robust and locally efficient estimates of the variable importance parameters and inference based on the influence curve. We demonstrate these properties through simulation under correct and incorrect model specification, and apply our method in practice to estimating the activity of transcription factor (TF) over cell cycle in yeast. We specifically target the importance of SWI4, SWI6, MBP1, MCM1, ACE2, FKH2, NDD1, and SWI5. The semiparametric model allows us to determine the importance of a TF at specific time points by specifying time indicators as potential effect modifiers of the TF. Our results are promising, showing significant importance trends during the expected time periods. This methodology can also be used as a variable importance analysis tool to assess the effect of a large number of variables such as gene expressions or single nucleotide polymorphisms.


A multidisciplinary, minimally invasive approach combining lacrimoscopy and fluoroscopically guided stenting for management of nasolacrimal apparatus obstruction in dogs.
OBJECTIVE To describe and evaluate outcomes of a multidisciplinary, minimally invasive approach combining lacrimoscopy and fluoroscopically guided stenting for management of nasolacrimal apparatus (NLA) obstruction in dogs. DESIGN Prospective, nonrandomized clinical trial. ANIMALS 16 client-owned dogs with confirmed NLA obstruction. PROCEDURES Dogs underwent CT contrast dacryocystorhinography, rhinoscopy, and lacrimoscopy. Whenever possible, the NLA was stented, typically with fluoroscopic guidance. RESULTS Median duration of clinical signs prior to treatment was 3.2 months (range, 0.2 to 14 months). Causes of NLA obstruction were a foreign body (n = 5), dacryocystitis (4), stenosis secondary to fibrosis (3), granulation tissue (1), or granulation tissue in association with a small foreign body (1); a cause was not identified in 2 dogs. Stents were placed in 14 of 16 (88%) dogs for a median duration of 5.6 weeks (range, 1.3 to 9.4 weeks). Stenting was not possible in 2 dogs with stenosis of the NLA secondary to granulation tissue or fibrosis. Owners of all 16 dogs reported at least 60% clinical improvement with median improvement rated as 95%, and owners of 8 dogs reporting complete resolution of signs. Two dogs required antimicrobial administration because of dacryocystitis that persisted after stent removal; a foreign body was not found in either dog. CONCLUSIONS AND CLINICAL RELEVANCE Overall clinical response and owner-rated improvement for dogs with NLA obstruction that underwent lacrimoscopy and fluoroscopically guided stenting were high, especially given that these dogs had failed to respond to conventional treatment.


Heat shock protein 70 neutralizes apoptosis-inducing factor.
Programmed cell death (apoptosis) is the physiological process responsible for the demise of superfluous, aged, damaged, mutated, and ectopic cells. Its normal function is essential both for embryonic development and for maintenance of adult tissue homeostasis. Deficient apoptosis participates in cancerogenesis, whereas excessive apoptosis leads to unwarranted cell loss accounting for disparate diseases including neurodegeneration and AIDS. One critical step in the process of apoptosis consists in the permeabilization of mitochondrial membranes, leading to the release of proteins which normally are secluded behind the outer mitochondrial membrane. For example, cytochrome c, which is normally confined to the mitochondrial intermembrane space, is liberated from mitochondria and interacts with a cytosolic protein, Apaf-1, causing its oligomerization and constitution of the so-called apoptosome, a protein complex which activates a specific class of cysteine proteases, the caspases[2]. Another example concerns the so-called apoptosis-inducing factor (AIF), another mitochondrial intermembrane protein which can translocate to the nucleus where it induces chromatin condensation and DNA fragmentation.


Anxiety sensitivity, panic, and depressed mood: a reanalysis teasing apart the contributions of the two levels in the hierarchial structure of the Anxiety Sensitivity Index.
Anxiety sensitivity (AS) has been defined as the fear of anxiety and anxiety-related sensations, and evidence suggests that AS plays an important role in the psychopathology of panic. It is entirely unclear whether the relation between AS and panic should be attributed to one (or more) of the AS group factors, the general AS factor, or to factors at both levels of the AS hierarchy. The authors reanalyzed data presented earlier by R. M. Rapee, T. A. Brown, M. A. Antony, and D. H. Barlow (1992) to tease apart the contributions of the different levels of the AS hierarchy to fear responses to hyperventilation and 5.5% carbon dioxide challenges. The results demonstrated that AS-Physical Concerns is the only one of the three AS group factors that contributes to relations with fear responses to these two challenges. However, AS-Mental Incapacitation Concerns had a stronger positive linear association with depressed mood than did AS-Physical Concerns.