Living on the edge: roads and edge effects on small mammal populations.
1. Roads may affect wildlife populations through habitat loss and disturbances, as they create an abrupt linear edge, increasing the proportion of edge exposed to a different habitat. Three types of edge effects have been recognized: abiotic, direct biotic, and indirect biotic. 2. We explored the direct biotic edge effects of 3- to 4-m wide roads, and also a previously unrecognized type of edge effect: social. We live-trapped two threatened endemic rodents from Cozumel Island (Oryzomys couesi cozumelae and Reithrodontomys spectabilis) in 16 plots delimited by roads on two sides, to compare edge effects between two adjacent edges (corners), single-edge and interior forest, on life history and social variables. 3. No significant edge effects were observed on the life-history variables, with the exception of differences in body condition between males and females of O. c. cozumelae near edges. Both species showed significant and contrasting effects on their social variables. 4. O. c. cozumelae was distributed according to its age and sex: the proportion of adults and males was higher in interior than near edges, while juveniles and females were more abundant near edges. More nonreproductive females were present in corners than in single-edge and interior, while the opposite distribution was observed for nonreproductive males. 5. The distribution of R. spectabilis was related to its age and reproductive condition, but not to its sex. The proportion of adults was significantly higher in corners, while juveniles were only caught in single-edge and interior quadrants. The proportion of reproductive individuals was higher in edge than interior quadrants, while reproductive females were only present in edge quadrants. 6. We found significant differences between the quadrants with the greatest edge exposure in comparison with other quadrants. The social edge effects we identified complement the typology of edge effects recognized in ecological literature. Our study provides insight into the effects that sharp road edges have on biological and social characteristics of small mammal populations, highlighting how such effects vary among species. Our findings have important conservation implications for these threatened species, but are also applicable in a broader context wherever there are abrupt edges caused by linear landscape features.


Testing the inhibitory cascade model in a recent human sample.
The Inhibitory Cascade Model was proposed by Kavanagh and colleagues (Nature, 449, 427-433 [2007]) after their experimental studies on the dental development of murine rodent species. These authors described an activator-inhibitor mechanism that has been employed to predict evolutionary size patterns of mammalian teeth, including hominins. In the present study, we measured the crown area of the three lower permanent molars (M1, M2, and M3) of a large recent modern human sample of male and female individuals from a collection preserved at the Institute of Anthropology of the University of Coimbra (Portugal). The main aim of the present study is to test if the size molar patterns observed in this human sample fits the Inhibitory Cascade Model. For this purpose, we first measured the crown area in those individuals preserving the complete molar series. Measurements were taken in photographs, using a planimeter and following well-tested techniques used in previous works. We then plot the M3 /M1 and M2 /M1 size ratios. Our results show that the premise of the Inhibitory Cascade Model, according to which the average of the crown area of M2 is approximately one-third of the sum of the crown area of the three molars, is fulfilled. However, our results also show that the individual values of a significant number of males and females are out of the 95% confidence interval predicted by the Inhibitory Cascade Model in rodents. As a result, the present analyses suggest that neither the sample of males, nor that of females, nor the pooled sample fits the Inhibitory Cascade Model. It is important to notice that, although this model has been successfully tested in a large number of current human populations, to the best of our knowledge this is the first study in which individual data have been obtained in a recent human population rather than using the average of the sample. Our results evince that, at the individual level, some factors not yet known could interfere with this model masking the modulation of the size on the molar series in modern humans. We suggest that the considerable delay in the onset of M3 formation in modern humans could be related to a weakening of the possible activation/inhibition process for this tooth. Finally, and in support of our conclusions, we have checked that the absolute and relative size of M1 and M2 is not related to the M3 agenesis in our sample. In line with other studies in primates, our results do not support the Inhibitory Cascade Model in a recent human sample. Further research is needed to better understand the genetic basis of this mechanism and its relationship to the phenotype. In this way, we may be able to find out which evolutionary changes may be responsible for the deviations observed in many species, including Homo sapiens.


Fabrication of multilayer mirrors consisting of oxide and nitride layers for continual use across the K-absorption edge of carbon.
The development of multilayer mirrors for continual use around the K-absorption edge of carbon (4.4 nm) has been begun. Cobalt oxide (Co3O4), silicon oxide (SiO2), and boron nitride (BN) are found to be suitable for multilayer mirrors on the basis of theoretical calculations for wavelengths around the carbon K-absorption edge region. X-ray reflectivity curves with CuKalpha1 x rays of the fabricated Co3O4/SiO2 multilayers have sharp Bragg peaks, and the layer structures evaluated from transmission electron microscopy (TEM) observations are uniform. On the other hand, the Bragg peaks of Co3O4/BN multilayers split, and aggregated Co3O4 is observed. To improve the Co3O4 layer structure, chromium oxide (Cr2O3) was mixed into Co3O4. The mixed oxide layer structure in the Mix/BN multilayer (Mix = Co3O4 + Cr2O3) is relatively uniform, and the Bragg peaks do not split.


Inter- and intrarater reliability of the Chicago Classification in pediatric high-resolution esophageal manometry recordings.
The Chicago Classification (CC) facilitates interpretation of high-resolution manometry (HRM) recordings. Application of this adult based algorithm to the pediatric population is unknown. We therefore assessed intra and interrater reliability of software-based CC diagnosis in a pediatric cohort. Thirty pediatric solid state HRM recordings (13M; mean age 12.1 ± 5.1 years) assessing 10 liquid swallows per patient were analyzed twice by 11 raters (six experts, five non-experts). Software-placed anatomical landmarks required manual adjustment or removal. Integrated relaxation pressure (IRP4s), distal contractile integral (DCI), contractile front velocity (CFV), distal latency (DL) and break size (BS), and an overall CC diagnosis were software-generated. In addition, raters provided their subjective CC diagnosis. Reliability was calculated with Cohen's and Fleiss' kappa (κ) and intraclass correlation coefficient (ICC). Intra- and interrater reliability of software-generated CC diagnosis after manual adjustment of landmarks was substantial (mean κ = 0.69 and 0.77 respectively) and moderate-substantial for subjective CC diagnosis (mean κ = 0.70 and 0.58 respectively). Reliability of both software-generated and subjective diagnosis of normal motility was high (κ = 0.81 and κ = 0.79). Intra- and interrater reliability were excellent for IRP4s, DCI, and BS. Experts had higher interrater reliability than non-experts for DL (ICC = 0.65 vs ICC = 0.36 respectively) and the software-generated diagnosis diffuse esophageal spasm (DES, κ = 0.64 vs κ = 0.30). Among experts, the reliability for the subjective diagnosis of achalasia and esophageal gastric junction outflow obstruction was moderate-substantial (κ = 0.45-0.82). Inter- and intrarater reliability of software-based CC diagnosis of pediatric HRM recordings was high overall. However, experience was a factor influencing the diagnosis of some motility disorders, particularly DES and achalasia.


Type III functional response in Daphnia.
The functional response of Daphnia, a common pelagic herbivore in lakes, was assessed with a combination of secondary and meta-analyses of published data and new data from an experiment conducted using very low food levels. Secondary analyses of literature data (28 studies, n = 239-393) revealed a significant positive influence of food concentration on Daphnia clearance rate at low food levels, i.e., evidence of an overall Type III functional response. This result was not an artifact of including data from Daphnia that were exhausted from prolonged food deprivation (more than three hours at very low food). Meta-analysis of Daphnia clearance rate vs. food concentration across a range of low food concentrations (eight studies) showed a significantly positive slope across studies, which also supports the presence of a Type III response. Congruent with these analyses of published data, the feeding experiment showed clear evidence of a Type III functional response for D. pulicaria feeding on Ankistrodesmus falcatus. Food levels at which Daphnia clearance rate declined with decreasing food were near the minimum resource requirement for Daphnia population maintenance at steady state (R*). We suggest that Type III responses are more common than previously believed, perhaps because of the relative paucity of observations at low food levels, and that reduced prey mortality at low phytoplankton densities could be a stabilizing mechanism for Daphnia-phytoplankton systems under resource scarcity.


Health-related quality of life in patients with prominent negative symptoms: results from a multicenter randomized Phase II trial on bitopertin.
Symptoms of schizophrenia fall into three categories (positive, negative and cognitive symptoms), which probably impact differently on patient's health-related quality of life (HRQoL). The present study aimed to explore HRQoL in patients with prominent negative symptoms. In the 323 patients with prominent negative symptoms included in a multicenter Phase II trial investigating the safety and efficacy of bitopertin, HRQoL was assessed using the Schizophrenia Quality of Life Scale (SQLS), symptoms severity using the Positive and Negative Syndrome Scale and functioning using the Personal and Social Performance Scale. SQLS measurement properties were assessed; HRQoL was compared between treatment arms, and relationships between HRQoL, symptoms and functioning at baseline were explored. Both SQLS scores (Vitality/Cognition and Psychosocial Feelings) demonstrated good test-retest (ICC = 0.77 and 0.74) and internal consistency reliability (Cronbach's α = 0.86 and 0.93). Clinical validity with regard to schizophrenia severity and ability to detect change in severity of symptoms of schizophrenia were satisfactory. The SQLS structure was not formally disconfirmed. No statistically significant difference was observed between treatment arms. Negative symptoms were more strongly associated with functioning than positive symptoms. Functioning and Anxiety/Depression were strongly related to both SQLS domains. Overall, SQLS measurement properties were supported in these patients with prominent negative symptoms of schizophrenia. The impact of negative symptoms on functioning and HRQoL suggests that improving these symptoms will be a meaningful benefit in this population of patients.


Cloning and characterization of the murine pkd2 promoter.
Pkd2, the mouse homologue of PKD2, the gene responsible for the second form of autosomal dominant polycystic kidney disease, is highly expressed in fetal and adult mouse tissues. The expression of Pkd2 is developmentally regulated. To begin to dissect out the regulatory mechanism of Pkd2 expression, we characterized the basic features of the gene structure and identified potential cis-regulatory elements of Pkd2 transcription. Pkd2 spans 42 kb with a transcription start site 165 bp upstream of the translation start codon. Exon 1 of Pkd2 is 755 bp long, and the full-length transcript is 5215 bp. The Pkd2 promoter region is GC-rich and lacks a consensus TATA or CCAAT box. Consensus binding sites for the transcription factors Sp-1, NF-1, and Ap-2 lie in the 5' upstream region of Pkd2. The Sp-1 binding site is conserved in 5' upstream sequences of both the mouse and the human genes. The CAT activity of a series of upstream segments from +178 to -2749 was assessed in MDCK, LLCPK1, COS-7, and HEK293 cells. Deletion analysis identified a 409-bp fragment from position -221 to +178 responsible for basal promoter activity. A 922-bp fragment from -744 to +178 showed the highest level of CAT activity in the cell lines tested. These data define a functional promoter candidate region for Pkd2.


The paired box encodes a second DNA-binding domain in the paired homeo domain protein.
The homeo box, which encodes the DNA-binding homeo domain, is a DNA sequence motif present in several Drosophila developmental genes; it has been used to identify many homologous genes involved in mammalian development. The paired box is another conserved sequence motif, first identified in the paired (prd) and gooseberry (gsb) Drosophila homeo domain genes. It encodes a 128-amino-acid domain, the paired domain, which has since been found in other fly and mouse gene products, in association with the homeo domain or in its absence. We show that the paired box of the prd gene encodes a DNA-binding activity, independent of the DNA-binding activity of the Paired (Prd) homeo domain and with a different sequence specificity. The amino-terminal region of the paired domain, including one of the three predicted alpha-helices, is necessary and sufficient for binding. We investigate the binding of the Prd protein to two sites in the even-skipped promoter, which are composed of overlapping sequences bound by the homeo domain and by the paired domain. We also show that a mutation in the paired box of Prd, corresponding to the mutation in the paired box of the mouse Pax-1 gene thought to cause the undulated skeletal phenotype, destroys the ability of the Prd protein to bind to the paired domain-specific site. This supports the view that the undulated phenotype results from the inactivation of the DNA-binding activity of the paired domain of Pax-1.


Repeatability of corneal parameters with Pentacam after laser in situ keratomileusis.
To investigate the coefficient of repeatability (CR) for corneal parameters evaluated with Pentacam after laser in situ keratomileusis (LASIK) in myopic eyes. Prospective, non-interventional, non-comparative study in an institutional setup. Forty eyes of 40 consecutive subjects who had undergone LASIK for myopia were assessed with the Scheimpflug system (Pentacam 70700: Oculus, Wetzlar Germany). The mean of five consecutive measurements of all the corneal parameters was recorded and CR was calculated as standard deviation of the difference from the mean of these repeat measurements divided by the mean response. The statistical significance of the CR was calculated for these parameters at 5% significance level. The best CR was observed for the periphery of the anterior corneal curvature (0.18%) and the least for the horizontal meridian of the posterior corneal curvature (1.29%). Despite being significantly different ( P < 0.001), both the measurements were highly repeatable in post-LASIK eyes. The central, apical and minimal corneal thickness had a CR of 1%, 0.78% and 0.77% respectively. These were equally repeatable ( P> 0.323). The CR of the mean radius of curvature of the anterior cornea (0.29%) was significantly better ( P < 0.001) than the posterior corneal curvature (0.57%). The CR for the post-LASIK cornea with Pentacam was the best for the anterior corneal curvature. Significantly, Pentacam has a high degree of repeatability for the posterior corneal curvature, which has a potential for early detection of keratectasia in these eyes. Post-LASIK pachymetry with Pentacam also showed excellent repeatability.


ωB97X-V: a 10-parameter, range-separated hybrid, generalized gradient approximation density functional with nonlocal correlation, designed by a survival-of-the-fittest strategy.
A 10-parameter, range-separated hybrid (RSH), generalized gradient approximation (GGA) density functional with nonlocal correlation (VV10) is presented. Instead of truncating the B97-type power series inhomogeneity correction factors (ICF) for the exchange, same-spin correlation, and opposite-spin correlation functionals uniformly, all 16,383 combinations of the linear parameters up to fourth order (m = 4) are considered. These functionals are individually fit to a training set and the resulting parameters are validated on a primary test set in order to identify the 3 optimal ICF expansions. Through this procedure, it is discovered that the functional that performs best on the training and primary test sets has 7 linear parameters, with 3 additional nonlinear parameters from range-separation and nonlocal correlation. The resulting density functional, ωB97X-V, is further assessed on a secondary test set, the parallel-displaced coronene dimer, as well as several geometry datasets. Furthermore, the basis set dependence and integration grid sensitivity of ωB97X-V are analyzed and documented in order to facilitate the use of the functional.


Sensitivity of mottled sculpins (Cottus bairdi) and rainbow trout (Onchorhynchus mykiss) to acute and chronic toxicity of cadmium, copper, and zinc.
Studies of fish communities of streams draining mining areas suggest that sculpins (Cottus spp.) may be more sensitive than salmonids to adverse effects of metals. We compared the toxicity of zinc, copper, and cadmium to mottled sculpin (C. bairdi) and rainbow trout (Onchorhynchus mykiss) in laboratory toxicity tests. Acute (96-h) and early life-stage chronic (21- or 28-d) toxicity tests were conducted with rainbow trout and with mottled sculpins from populations in Minnesota and Missouri, USA, in diluted well water (hardness = 100 mg/L as CaCO3). Acute and chronic toxicity of metals to newly hatched and swim-up stages of mottled sculpins differed between the two source populations. Differences between populations were greatest for copper, with chronic toxicity values (ChV = geometric mean of lowest-observed-effect concentration and no-observed-effect concentration) of 4.4 microg/L for Missouri sculpins and 37 microg/L for Minnesota sculpins. Cadmium toxicity followed a similar trend, but differences between sculpin populations were less marked, with ChVs of 1.1 microg/L (Missouri) and 1.9 microg/L (Minnesota). Conversely, zinc was more toxic to Minnesota sculpins (ChV = 75 microg/L) than Missouri sculpins (chronic ChV = 219 microg/L). Species-average acute and chronic toxicity values for mottled sculpins were similar to or lower than those for rainbow trout and indicated that mottled sculpins were among the most sensitive aquatic species to toxicity of all three metals. Our results indicate that current acute and chronic water quality criteria for cadmium, copper, and zinc adequately protect rainbow trout but may not adequately protect some populations of mottled sculpins. Proposed water quality criteria for copper based on the biotic ligand model would be protective of both sculpin populations tested.


Origin of the overpotentials for HCOO- and CO formation in the electroreduction of CO2 on Cu(211): the reductive desorption processes decide.
Electroreduction of CO2 on Cu surface provides the potential in producing hydrocarbons and other multi-carbon products. However, a comprehensive understanding of the potential-related mechanism is required to improve the product selectivity as well as to reduce the overpotentials. Herein, we systematically characterize the potential effect on the complete reaction pathways to CO and HCOO- on the Cu(211) surface. Reaction free energy and activation barrier are computed as functions of electric potential. It is found that chemical adsorption state of CO2 is effectively stabilized by the substrate, which is expected to be dominant at potentials below -0.27 V vs. SHE, much earlier than that previously reported on Cu(100). Considering that the activation barriers of the other surface processes are small enough to be overcome at room temperature, the large reductive desorption free energies of OH- and HCOO- are suggested as the origin of high overpotentials.


Interventions for preventing oral mucositis in patients with cancer receiving treatment: oral cryotherapy.
Oral mucositis is a side effect of chemotherapy, head and neck radiotherapy, and targeted therapy, affecting over 75% of high risk patients. Ulceration can lead to severe pain and difficulty eating and drinking, which may necessitate opioid analgesics, hospitalisation and nasogastric or intravenous nutrition. These complications may lead to interruptions or alterations to cancer therapy, which may reduce survival. There is also a risk of death from sepsis if pathogens enter the ulcers of immunocompromised patients. Ulcerative oral mucositis can be costly to healthcare systems, yet there are few preventive interventions proven to be beneficial. Oral cryotherapy is a low-cost, simple intervention which is unlikely to cause side-effects. It has shown promise in clinical trials and warrants an up-to-date Cochrane review to assess and summarise the international evidence. To assess the effects of oral cryotherapy for preventing oral mucositis in patients with cancer who are receiving treatment. We searched the following databases: the Cochrane Oral Health Group Trials Register (to 17 June 2015), the Cochrane Central Register of Controlled Trials (CENTRAL) (Cochrane Library 2015, Issue 5), MEDLINE via Ovid (1946 to 17 June 2015), EMBASE via Ovid (1980 to 17 June 2015), CANCERLIT via PubMed (1950 to 17 June 2015) and CINAHL via EBSCO (1937 to 17 June 2015). We searched the US National Institutes of Health Trials Registry, and the WHO Clinical Trials Registry Platform for ongoing trials. No restrictions were placed on the language or date of publication when searching databases. We included parallel-design randomised controlled trials (RCTs) assessing the effects of oral cryotherapy in patients with cancer receiving treatment. We used outcomes from a published core outcome set registered on the COMET website. Two review authors independently screened the results of electronic searches, extracted data and assessed risk of bias. We contacted study authors for information where feasible. For dichotomous outcomes, we reported risk ratios (RR) and 95% confidence intervals (CI). For continuous outcomes, we reported mean differences (MD) and 95% CIs. We pooled similar studies in random-effects meta-analyses. We reported adverse effects in a narrative format. We included 14 RCTs analysing 1280 participants. The vast majority of participants did not receive radiotherapy to the head and neck, so this review primarily assesses prevention of chemotherapy-induced oral mucositis. All studies were at high risk of bias. The following results are for the main comparison: oral cryotherapy versus control (standard care or no treatment). Adults receiving fluorouracil-based (5FU) chemotherapy for solid cancersOral cryotherapy probably reduces oral mucositis of any severity (RR 0.61, 95% CI 0.52 to 0.72, 5 studies, 444 analysed, moderate quality evidence). In a population where 728 per 1000 would develop oral mucositis, oral cryotherapy would reduce this to 444 (95% CI 379 to 524). The number needed to treat to benefit one additional person (NNTB), i.e. to prevent them from developing oral mucositis, is 4 people (95% CI 3 to 5).The results were similar for moderate to severe oral mucositis (RR 0.52, 95% CI 0.41 to 0.65, 5 studies, 444 analysed, moderate quality evidence). NNTB 4 (95% CI 4 to 6).Severe oral mucositis is probably reduced (RR 0.40, 95% CI 0.27 to 0.61, 5 studies, 444 analysed, moderate quality evidence). Where 300 per 1000 would develop severe oral mucositis, oral cryotherapy would reduce this to 120 (95% CI 81 to 183), NNTB 6 (95% CI 5 to 9). Adults receiving high-dose melphalan-based chemotherapy before haematopoietic stem cell transplantation (HSCT)Oral cryotherapy may reduce oral mucositis of any severity (RR 0.59, 95% CI 0.35 to 1.01, 5 studies, 270 analysed, low quality evidence). Where 824 per 1000 would develop oral mucositis, oral cryotherapy would reduce this to 486 (95% CI reduced to 289 to increased to 833). The NNTB is 3, although the uncertainty surrounding the effect estimate means that the 95% CI ranges from 2 NNTB, to 111 NNTH (number needed to treat in order to harm one additional person, i.e. for one additional person to develop oral mucositis).The results were similar for moderate to severe oral mucositis (RR 0.43, 95% CI 0.17 to 1.09, 5 studies, 270 analysed, low quality evidence). NNTB 3 (95% CI 2 NNTB to 17 NNTH).Severe oral mucositis is probably reduced (RR 0.38, 95% CI 0.20 to 0.72, 5 studies, 270 analysed, moderate quality evidence). Where 427 per 1000 would develop severe oral mucositis, oral cryotherapy would reduce this to 162 (95% CI 85 to 308), NNTB 4 (95% CI 3 to 9).Oral cryotherapy was shown to be safe, with very low rates of minor adverse effects, such as headaches, chills, numbness/taste disturbance, and tooth pain. This appears to contribute to the high rates of compliance seen in the included studies.There was limited or no evidence on the secondary outcomes of this review, or on patients undergoing other chemotherapies, radiotherapy, targeted therapy, or on comparisons of oral cryotherapy with other interventions or different oral cryotherapy regimens. Therefore no further robust conclusions can be made. There was also no evidence on the effects of oral cryotherapy in children undergoing cancer treatment. We are confident that oral cryotherapy leads to large reductions in oral mucositis of all severities in adults receiving 5FU for solid cancers. We are less confident in the ability of oral cryotherapy to reduce oral mucositis in adults receiving high-dose melphalan before HSCT. Evidence suggests that it does reduce oral mucositis in these adults, but we are less certain about the size of the reduction, which could be large or small. However, we are confident that there is an appreciable reduction in severe oral mucositis in these adults.This Cochrane review includes some very recent and currently unpublished data, and strengthens international guideline statements for adults receiving the above cancer treatments.


Oxidative tryptophan metabolism in renal allograft recipients: increased kynurenine synthesis is associated with inflammation and OKT3 therapy.
Serum concentrations of tryptophan (TRP) and kynurenine (KYN) were determined in renal allograft recipients (RAR) as an index of interferon-gamma-induced, indoleamine-dioxygenase-catalysed TRP degradation. Serum TRP and KYN in RAR during periods of stable graft function were typically within the normal range, however, the median values for serum KYN demonstrated significant increases 5-7 days prior to biopsy-confirmed acute rejection (1.6-fold, P less than 0.01) and on the day of biopsy (1.7-fold, P less than 0.001). Serum KYN was also markedly elevated in patients who contracted viral or Gram-negative bacterial infections in the absence of graft rejection. Serum KYN was not correlated with serum creatinine in RAR nor were serum TRP or KYN affected by antirejection therapy with high dose steroids. Retrospective analysis of intra-patient changes in serum KYN demonstrated that KYN monitoring was a useful adjunct to serum creatinine in the early detection of first acute rejection episodes. The first course of OKT3 therapy was associated with low serum TRP and significant increases in serum KYN (two- to three-fold) following the first three doses. The time course of these abnormalities corresponded to that over which many of the side effects of the OKT3 'first dose reaction' have been reported to occur. Significant changes in serum KYN were not observed in patients receiving repeat courses of OKT3 therapy. Significant decreases in serum TRP and significant increases in serum KYN were both prevalent and frequent in RAR during the first two postoperative months.(ABSTRACT TRUNCATED AT 250 WORDS)


Quantum optics with particles of light.
The concept of particles of light is introduced by ascribing a mechanical degree of freedom to a radiational wave packet. Evolution of the position and momentum observables of such particle in a dispersive dielectric is studied. It is shown that an initial coherent state evolves into a dispersed state which is characterized by a reduction of quantum fluctuations below Standard Quantum Limit when a certain combination of the position and momentum observables is measured.


Hybrid photocathode consisting of a CuGaO2 p-type semiconductor and a Ru(ii)-Re(i) supramolecular photocatalyst: non-biased visible-light-driven CO2 reduction with water oxidation.
A CuGaO2 p-type semiconductor electrode was successfully employed for constructing a new hybrid photocathode with a Ru(ii)-Re(i) supramolecular photocatalyst (RuRe/CuGaO2). The RuRe/CuGaO2 photocathode displayed photoelectrochemical activity for the conversion of CO2 to CO in an aqueous electrolyte solution with a positive onset potential of +0.3 V vs. Ag/AgCl, which is 0.4 V more positive in comparison to a previously reported hybrid photocathode that used a NiO electrode instead of CuGaO2. A photoelectrochemical cell comprising this RuRe/CuGaO2 photocathode and a CoO x /TaON photoanode enabled the visible-light-driven catalytic reduction of CO2 using water as a reductant to give CO and O2 without applying any external bias. This is the first self-driven photoelectrochemical cell constructed with the molecular photocatalyst to achieve the reduction of CO2 by only using visible light as the energy source and water as a reductant.


A single-nucleotide polymorphism in the EAP1 gene is associated with amenorrhea/oligomenorrhea in nonhuman primates.
Current evidence suggests that the acquisition of female reproductive capacity and the maintenance of mature reproductive function are related processes transcriptionally regulated by gene networks operating within the neuroendocrine brain. One of these genes, termed enhanced at puberty 1 (EAP1), encodes an upstream regulator of these processes. Selective inhibition of EAP1 expression in discrete regions of the rat and nonhuman primate (NHP) hypothalamus, via targeted delivery of RNA interference, either disrupts (rats) or abolishes (monkeys) reproductive cycles. The striking loss of menstrual cyclicity resulting from knocking down hypothalamic EAP1 expression suggests that diminished EAP1 function may contribute to disorders of the menstrual cycle of neuroendocrine origin. Here we show that a single-nucleotide polymorphism in the 5'-flanking region of EAP1 gene is associated with increased incidence of amenorrhea/oligomenorrhea in NHP. In the presence of the risk allele, binding of the transcription factor mothers against decapentaplegic homolog 3 (SMAD3) to its recognition site contained within the polymorphic sequence in the monkey EAP1 promoter is reduced. The risk allele also diminishes the increase in EAP1 promoter activity elicited by TGFβ1, a peptide that activates a SMAD3/4-mediated signaling pathway to regulate gene transcription. These findings indicate that common genetic variation in the EAP1 locus increases the susceptibility of NHP to loss/disruption of menstrual cyclicity. They also raise the possibility that polymorphisms in EAP1 may increase the risk of functional hypothalamic amenorrhea in humans.


Mechanism of toxic action of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in cultured human luteinized granulosa cells.
2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) caused a significant decrease in estradiol (E2) production when it was administered to human luteinized granulosa cells (hLGCs) in culture. We investigated the involvement of the epidermal growth factor receptor (EGFR) and protein tyrosine kinase (PTK) in this TCDD-induced toxicity. Upregulation in 125I-EGF binding to EGFR was measured after 24 h of TCDD treatment, while downregulation in EGFR binding was measured after 72 h of TCDD treatment. Upregulation of EGFR binding was associated with a significant decrease in postnuclear (7000 x g supernatant) PTK activity, but this activity was stimulated after 72 h of TCDD treatment. TCDD altered the level of tyrosine phosphorylation in proteins with molecular weights 35, 40, 43, 45, 60, and > 205 kDa. TCDD caused a significant increase in postnuclear cAMP-dependent protein kinase (PKA) after 24 h of treatment. The actions of TCDD on protein kinases were partially blocked by the protein synthesis inhibitor, cycloheximide. On the other hand, TCDD increased nuclear PTK and decreased nuclear PKA activity. E2 inhibited the postnuclear and nuclear activity of both PTK and PKA in control samples, but did not affect TCDD actions on either postnuclear or nuclear PTK activity. However, E2 abolished the stimulatory effect of TCDD on PKA activity in postnuclear protein. In the presence of insulin, TCDD did not induce any additional changes in postnuclear or nuclear PTK. Forskolin (FK) alone inhibited postnuclear PTK activity and stimulated its nuclear activity. The addition of TCDD 20 min after FK resulted in an increase in postnuclear PTK, but there was little change in nuclear PTK as compared to the effect of FK alone. The stimulatory effect of TCDD on postnuclear PKA activity was enhanced by insulin and TCDD reversed the negative effect of FK, but there was no effect of either insulin or FK on the inhibition by TCDD of nuclear PKA activity. TCDD decreased the activity of MAP2 kinase and reduced the binding activity of AP-1 DNA when given alone, and also blocked the E2 stimulation of MAP2K. These findings suggest that TCDD may interrupt the endocrine function of hLGCs through the blockage of the mitotic signal directly or indirectly through the interaction of PTK/MAP2K and PKA signaling.


Most published meta-regression analyses based on aggregate data suffer from methodological pitfalls: a meta-epidemiological study.
Due to clinical and methodological diversity, clinical studies included in meta-analyses often differ in ways that lead to differences in treatment effects across studies. Meta-regression analysis is generally recommended to explore associations between study-level characteristics and treatment effect, however, three key pitfalls of meta-regression may lead to invalid conclusions. Our aims were to determine the frequency of these three pitfalls of meta-regression analyses, examine characteristics associated with the occurrence of these pitfalls, and explore changes between 2002 and 2012. A meta-epidemiological study of studies including aggregate data meta-regression analysis in the years 2002 and 2012. We assessed the prevalence of meta-regression analyses with at least 1 of 3 pitfalls: ecological fallacy, overfitting, and inappropriate methods to regress treatment effects against the risk of the analysed outcome. We used logistic regression to investigate study characteristics associated with pitfalls and examined differences between 2002 and 2012. Our search yielded 580 studies with meta-analyses, of which 81 included meta-regression analyses with aggregated data. 57 meta-regression analyses were found to contain at least one pitfall (70%): 53 were susceptible to ecological fallacy (65%), 14 had a risk of overfitting (17%), and 5 inappropriately regressed treatment effects against the risk of the analysed outcome (6%). We found no difference in the prevalence of meta-regression analyses with methodological pitfalls between 2002 and 2012, nor any study-level characteristic that was clearly associated with the occurrence of any of the pitfalls. The majority of meta-regression analyses based on aggregate data contain methodological pitfalls that may result in misleading findings.


bcl-2 and c-erbB-2 proteins are involved in the regulation of VEGF and of thymidine phosphorylase angiogenic activity in non-small-cell lung cancer.
Tumour angiogenesis has been recently recognised as one of the most important prognostic factors in lung cancer. Although a variety of angiogenic factors have been identified, the angiogenesis process remains poorly understood. Bcl-2, c-erbB-2 and p53 are well-known oncogenes involved in non-small-cell lung cancer pathogenesis. A direct correlation of thymidine phosphorylase (TP) and of vascular endothelial growth factor (VEGF) with intratumoural angiogenesis has been reported. In the present study we investigated the possible regulatory role of bcl-2, c-erB-2 proteins in angiogenesis and in VEGF and TP expression in non-small-cell lung cancer. Two hundred sixteen specimens from T1,2-N0,1 staged patients treated with surgery alone were immunohistochemically examined. Bcl-2 and c-erbB-2 were significantly inversely related to each other (P = 0.04) and both were inversely associated with microvessel density (P < 0.02). High TP and VEGF reactivity was statistically related to loss of bcl-2 expression (P < 0.01). A significant co-expression of c-erbB-2 with TP was noted (P = 0.01). However, TP expression was related to high angiogenesis only in cases with absence of c-erB-2 expression (P < 0.0001). c-erbB-2 expression in poorly vascularised tumours was linked with poor outcome (P = 0.03). The present study provides strong evidence that the bcl-2 gene has a suppressive function over genes involved in both angiogenesis (VEGF and TP) and cell migration (c-erbB-2) in NSCLC. TP and c-erbB-2 proteins are significantly, and often simultaneously, expressed in bcl-2 negative cases. However, expression of the c-erbB-2 abolishes the TP-related angiogenic activity. Whether this is a result of a direct activity of the c-erbB-2 protein or a consequence of a c-erbB-2-related immune response remains to be further investigated.


Health Informatics Can Avoid Committing Symbolic Violence by Recognizing and Supporting Generic Decision-making Competencies.
'Symbolic violence' is committed, however well-intentionally, by the imposition of particular conceptualizations of what information, in what form and quality, is needed in order to make an 'informed choice' and hence - by questionable segue - a high quality decision. The social and cultural forms of relevant cognitive capital possessed by those who fail, because of their low general literacy, professionally-set knowledge tests of functional health literacy, are being ignored. Failing to recognise and exploit a particular form of functional decision literacy, in fact leads to symbolic violence being experienced by individuals at any and all levels of general literacy. It leads many to adopt the same range of avoidant and other undesirable strategies within healthcare situations observed in those of low basic literacy. The alternative response we propose exploits the alternative generic decision literacy which comes in the form of the ability to access and use the decision-relevant resources provided for many consumer services and products on comparison websites and magazines. The methodology is the simple form of multi-criteria analysis in which the products' ratings on multiple criteria are combined with criterion weights (supplied by the site) to produce scores and 'best buys' and 'good value for money' verdicts. Our alternative approach extends this approach to healthcare options and permits the incorporation of personal criterion weights in furtherance of person-centred care. Health informaticians, especially those in the decision support field, should build on this widespread generic competence. The fact that it is generic, far from implying context insensitivity, can be seen as a necessary basis for achieving context-sensitivity and sensitivisation at the level of the individual person as they experience a lifelong sequence of healthcare decisions.


Minor modifications to the phosphate groups and the C3' acyl chain length of lipid A in two Bordetella pertussis strains, BP338 and 18-323, independently affect Toll-like receptor 4 protein activation.
Lipopolysaccharides (LPS) of Bordetella pertussis are important modulators of the immune system. Interaction of the lipid A region of LPS with the Toll-like receptor 4 (TLR4) complex causes dimerization of TLR4 and activation of downstream nuclear factor κB (NFκB), which can lead to inflammation. We have previously shown that two strains of B. pertussis, BP338 (a Tohama I-derivative) and 18-323, display two differences in lipid A structure. 1) BP338 can modify the 1- and 4'-phosphates by the addition of glucosamine (GlcN), whereas 18-323 cannot, and 2) the C3' acyl chain in BP338 is 14 carbons long, but only 10 or 12 carbons long in 18-323. In addition, BP338 lipid A can activate TLR4 to a greater extent than 18-323 lipid A. Here we set out to determine the genetic reasons for the differences in these lipid A structures and the contribution of each structural difference to the ability of lipid A to activate TLR4. We show that three genes of the lipid A GlcN modification (Lgm) locus, lgmA, lgmB, and lgmC (previously locus tags BP0399-BP0397), are required for GlcN modification and a single amino acid difference in LpxA is responsible for the difference in C3' acyl chain length. Furthermore, by introducing lipid A-modifying genes into 18-323 to generate isogenic strains with varying penta-acyl lipid A structures, we determined that both modifications increase TLR4 activation, although the GlcN modification plays a dominant role. These results shed light on how TLR4 may interact with penta-acyl lipid A species.


Novel forms of forgetting in temporal lobe epilepsy.
Transient Epileptic Amnesia (TEA) is a recently defined subtype of temporal lobe epilepsy, principally affecting people in middle age with a male predominance. Its key manifestation is the occurrence of recurring episodes of transient amnesia, usually lasting less than an hour and often occurring on waking. One-third of patients have exclusively amnestic attacks, while in two-thirds, at least some attacks are accompanied by other manifestations of epilepsy, especially olfactory hallucinations. Several lines of evidence point to a seizure focus in the medial temporal lobes. Transient Epileptic Amnesia is accompanied by a striking loss of autobiographical memories in two-thirds of sufferers, accelerated loss of memories which had been acquired successfully in around one half, and topographical amnesia in around one-third. This paper reviews the findings of the TIME project (The Impairment of Memory in Epilepsy - http://sites.pcmd.ac.uk/time/tea.php) in relation to TEA, accelerated long-term forgetting, and remote memory impairment.


Microstimulation of the monkey superior colliculus induces pupil dilation without evoking saccades.
The orienting reflex is initiated by a salient stimulus and facilitates quick, appropriate action. It involves a rapid shift of the eyes, head, and attention and other physiological responses such as changes in heart rate and transient pupil dilation. The SC is a critical structure in the midbrain that selects incoming stimuli based on saliency and relevance to coordinate orienting behaviors, particularly gaze shifts, but its causal role in pupil dilation remains poorly understood in mammals. Here, we examined the role of the primate SC in the control of pupil dynamics. While requiring monkeys to keep their gaze fixed, we delivered weak electrical microstimulation to the SC, so that saccadic eye movements were not evoked. Pupil size increased transiently after microstimulation of the intermediate SC layers (SCi) and the size of evoked pupil dilation was larger on a dim versus bright background. In contrast, microstimulation of the superficial SC layers did not cause pupil dilation. Thus, the SCi is directly involved not only in shifts of gaze and attention, but also in pupil dilation as part of the orienting reflex, and the function of pupil dilation may be related to increasing visual sensitivity. The shared neural mechanisms suggest that pupil dilation may be associated with covert attention.


High spectral and spatial resolution MRI of age-related changes in murine prostate.
The purpose of this work was to evaluate high-resolution echo-planar spectroscopic MRI of normal and precancerous prostatic changes in a transgenic mouse line. Simian virus large T-antigen transgenic male mice (N = 7, age = 34 +/- 3.7 weeks) with prostatic hyperplasia and intraepithelial neoplasia (PIN) were studied. High spectral and spatial resolution (HiSS) MRI of the water proton signal was compared to the free induction decay (FID) integral image and conventional gradient-echo and spin-echo imaging. Water peak-height images of the prostate produced from HiSS datasets showed improved contrast-to-noise ratio (CNR) (P < 0.03), and greater morphological detail (P < 0.004) based on texture analysis. Despite the high spectral resolution of the HiSS datasets, signal-to-noise ratio (SNR) compared favorably with that of the FID integral and conventional images. Lobular features in HiSS images of older mice were consistent with hyperplasia seen on histology. A partially deuterated water-filled catheter was inserted in the mouse rectum for susceptibility matching between the colon interior and exterior to minimize image artifacts. These preliminary results suggest that HiSS MRI provides detailed morphology of the murine prostate and can detect early changes associated with the development of cancer. HiSS MRI of patients may have similar advantages.


Dinosaur energetics: setting the bounds on feasible physiologies and ecologies.
The metabolic status of dinosaurs has long been debated but remains unresolved as no consistent picture has emerged from a range of anatomical and isotopic evidence. Quantitative analysis of dinosaur energetics, based on general principles applicable to all vertebrates, shows that many features of dinosaur lifestyle are compatible with a physiology similar to that of extant lizards, scaled up to dinosaur body masses and temperatures. The analysis suggests that sufficient metabolic scope would have been available to support observed dinosaur growth rates and allow considerable locomotor activity, perhaps even migration. Since at least one dinosaur lineage evolved true endothermy, this study emphasizes there was no single dinosaur physiology. Many small theropods were insulated with feathers and appear to have been partial or full endotherms. Uninsulated small taxa, and all juveniles, presumably would have been ectothermic, with consequent diurnal and seasonal variations in body temperature. In larger taxa, inertial homeothermy would have resulted in warm and stable body temperatures but with a basal metabolism significantly below that of extant mammals or birds of the same size. It would appear that dinosaurs exhibited a range of metabolic levels to match the broad spectrum of ecological niches they occupied.


Isofuranodiene, the main volatile constituent of wild celery (Smyrnium olusatrum L.), protects d-galactosamin/lipopolysacchride-induced liver injury in rats.
Isofuranodiene is a natural sesquiterpene rich occurring in Smyrnium olusatrum, a forgotten culinary herb which was marginalised after the domestication of the improved form of celery. Our recent data showed that isofuranodiene inhibited the proliferation and induced apoptosis in cancer cells. In this study, we investigated its protective effect on d-galactosamine/lipopolysacchride (GalN/LPS)-induced liver injury in SD rats. Oral administration of isofuranodiene (20 and 50 mg/kg) dramatically inhibited GalN/LPS-induced serum elevation of aspartate aminotransferase, alanine aminotransferase and malondialdehyde levels, and significantly ameliorated liver injury as evidenced by the histological improvement in H&E staining. Furthermore, isofuranodiene treatment significantly inhibited GalN/LPS-induced mRNA expression of IL-1β, IL-6 and inducible nitric oxide synthase in liver tissues. The results from this study showed that isofuranodiene protects GalN/LPS-induced liver injury in SD rats and suggested that it may be a potential functional food ingredient for the prevention and treatment of liver diseases.


The consensus model for regulation of APRNs: Implications for nurse practitioners.
Purpose: To examine the Consensus Model for APRN Regulation as a proposed solution to simplify and unify regulation of advanced nursing practice with respect to licensure, certification, accreditation, and educational standards. Data sources: Policy statements and reports issued by national organizations of nursing professionals and discussion within the scholarly literature provided an overview of regulatory differences among the 50 U.S. states, their impacts on advanced practice registered nurse (APRN) practice, and discussion of the Consensus Model as a potential solution. Conclusions: Major organizations of nursing professionals have endorsed the Consensus Model, indicating the importance of this significant step toward standards and consistent quality in APRN education and practice. Diverse regulatory criteria, such as variable accreditation standards, disparate certification and licensing requirements, inconsistent population foci, and scopes of practice, represent barriers to optimized APRN function within today's complex healthcare system and, ultimately, reduced access to safe, quality care for patients. Implications for practice: The Consensus Model for APRN Regulation promises to unify APRN education, practice, and licensure, promoting greater mobility among nursing professionals. Elimination of inconsistencies in APRN licensure, education, certification, and accreditation requirements will promote greater value and mobility for the APRN within the healthcare system.


Withdrawal from treatment as an outcome in the ISOLDE study of COPD.
To investigate the determinants of patient withdrawal from our study, and the effect of these withdrawals on the outcome of treatment with inhaled corticosteroids in patients with COPD. A double-blind, placebo-controlled, randomized trial. Eighteen outpatient centers in the United Kingdom. Seven hundred fifty-one patients with stable COPD defined clinically and as baseline postbronchodilator FEV(1) > or = 0.8 L and < 85% predicted, FEV(1)/FVC ratio < 70%, and FEV(1) change after albuterol < 10% of predicted. Random assignment of either 500 microg bid of inhaled fluticasone propionate (FP) using a spacer device or an identical placebo inhaler. Treatment was continued for 3 years or until patients withdrew from follow-up. Postbronchodilator FEV(1) was measured on three occasions before randomization and every 3 months thereafter. Health status was assessed by the disease-specific St. George Respiratory Questionnaire (SGRQ) and the modified short-form 36 questionnaire (SF-36) at baseline and every 6 months. Three hundred thirty-nine patients withdrew, of whom 156 patients received FP. Prescription of frequent courses of oral prednisolone was the most common reason for withdrawing as specified in the protocol (69 patients in the FP group withdrew due to respiratory symptoms, compared with 93 patients in the placebo group). This explained the significantly greater dropout of placebo-treated patients that was most evident when FEV(1) was < 50% predicted. Patients withdrawing had a significantly more rapid decline in health status, measured by both the SGRQ and the SF-36 (p < 0.001). Those withdrawing from the placebo group had a more rapid decline in FEV(1) and more exacerbations than the FP-treated groups. Baseline FEV(1) was lower in dropouts than in patients completing the study receiving placebo, but there was no difference between the respective groups receiving FP. Patients who withdrew from follow-up were those with the most rapidly deteriorating health status and lung function. Losing these patients from the final analysis can reduce the power of a study to achieve its primary end point.


Can Hemp Help? Low-THC Cannabis and Non-THC Cannabinoids for the Treatment of Cancer.
Cannabis has been used to relieve the symptoms of disease for thousands of years. However, social and political biases have limited effective interrogation of the potential benefits of cannabis and polarised public opinion. Further, the medicinal and clinical utility of cannabis is limited by the psychotropic side effects of ∆9-tetrahydrocannabinol (∆9-THC). Evidence is emerging for the therapeutic benefits of cannabis in the treatment of neurological and neurodegenerative diseases, with potential efficacy as an analgesic and antiemetic for the management of cancer-related pain and treatment-related nausea and vomiting, respectively. An increasing number of preclinical studies have established that ∆9-THC can inhibit the growth and proliferation of cancerous cells through the modulation of cannabinoid receptors (CB1R and CB2R), but clinical confirmation remains lacking. In parallel, the anti-cancer properties of non-THC cannabinoids, such as cannabidiol (CBD), are linked to the modulation of non-CB1R/CB2R G-protein-coupled receptors, neurotransmitter receptors, and ligand-regulated transcription factors, which together modulate oncogenic signalling and redox homeostasis. Additional evidence has also demonstrated the anti-inflammatory properties of cannabinoids, and this may prove relevant in the context of peritumoural oedema and the tumour immune microenvironment. This review aims to document the emerging mechanisms of anti-cancer actions of non-THC cannabinoids.


Studying the joint effects of population stratification and sampling in case-control association studies.
Population stratification (PS) is referred to the systematic difference in allele frequencies between subpopulations in a population. It could cause a false-positive conclusion in a case-control association study, where the association is due to the structure of the underlying population, not a disease-associated locus. In this paper, we study the joint effects of PS and data sampling when the genetic effect is null. The level of the PS effect depends on the variation of the baseline genotype frequency across subpopulations and matching effectiveness of the sampling. In the case of simple random sampling (SRS), the matching effectiveness equals the inverse of the variation of the disease odds, and thus the PS bias is null under constant disease risk. However, if the latter condition holds but the sampling is not SRS, the bias may still exist. The magnitude of the bias increases as the deviation between the true sampling and SRS increases. We also derive bounds for the bias. If the bounds are approximately known or estimable, we show that this information can be used to compute a conservative p value for the usual association test. We give two real examples to demonstrate the application of the new method.


Overlapping functional anatomy for working memory and visual search.
Recent behavioural findings using dual-task paradigms demonstrate the importance of both spatial and non-spatial working memory processes in inefficient visual search (Anderson et al. in Exp Psychol 55:301-312, 2008). Here, using functional magnetic resonance imaging (fMRI), we sought to determine whether brain areas recruited during visual search are also involved in working memory. Using visually matched spatial and non-spatial working memory tasks, we confirmed previous behavioural findings that show significant dual-task interference effects occur when inefficient visual search is performed concurrently with either working memory task. Furthermore, we find considerable overlap in the cortical network activated by inefficient search and both working memory tasks. Our findings suggest that the interference effects observed behaviourally may have arisen from competition for cortical processes subserved by these overlapping regions. Drawing on previous findings (Anderson et al. in Exp Brain Res 180:289-302, 2007), we propose that the most likely anatomical locus for these interference effects is the inferior and middle frontal cortex of the right hemisphere. These areas are associated with attentional selection from memory as well as manipulation of information in memory, and we propose that the visual search and working memory tasks used here compete for common processing resources underlying these mechanisms.


T Cell Engineering.
T cells are a new and promising antigen-specific therapeutic option for the treatment of malignant diseases. To achieve antigen specificity against tumor antigens, T cells can be manipulated by gene transfer to express chimeric antigen receptors (CARs). CAR-expressing T cells are called redirected T cells. CARs are composed of an extracellular antibody-derived antigen recognition domain, a transmembrane domain and a cytoplasmatic signal domain. Therefore, redirected T cells combine the exchangeable specificity of an antibody with the cytotoxic machinery of a T cell. Early clinical trials with redirected T cells targeting cluster of differentiation (CD) 19 have shown impressive results in CD19-positive hematological cancers. However, for solid cancers only limited clinical experience exists and new and innovative concepts have to be developed to overcome tumor-mediated immune suppression. Herein, we describe the general design of a CAR, the function of the different domains and the different strategies to produce redirected T cells. Furthermore, we summarize and discuss the preclinical and clinical data indicating the tremendous potential of redirected T cells to become a mainstay of cancer immunotherapy.


Gcn5p-dependent acetylation induces degradation of the meiotic transcriptional repressor Ume6p.
Ume6p represses early meiotic gene transcription in Saccharomyces cerevisiae by recruiting the Rpd3p histone deacetylase and chromatin-remodeling proteins. Ume6p repression is relieved in a two-step destruction process mediated by the anaphase-promoting complex/cyclosome (APC/C) ubiquitin ligase. The first step induces partial Ume6p degradation when vegetative cells shift from glucose- to acetate-based medium. Complete proteolysis happens only upon meiotic entry. Here we demonstrate that the first step in Ume6p destruction is controlled by its acetylation and deacetylation by the Gcn5p acetyltransferase and Rpd3p, respectively. Ume6p acetylation occurs in medium lacking dextrose and results in a partial destruction of the repressor. Preventing acetylation delays Ume6p meiotic destruction and retards both the transient transcription program and execution of the meiotic nuclear divisions. Conversely, mimicking acetylation induces partial destruction of Ume6p in dextrose medium and accelerates meiotic degradation by the APC/C. These studies reveal a new mechanism by which acetyltransferase activity induces gene expression through targeted destruction of a transcriptional repressor. These findings also demonstrate an important role for nonhistone acetylation in the transition between mitotic and meiotic cell division.


Evaluation of strategies to reduce radiation dose in perfusion CT imaging using a reproducible biologic phantom.
The purpose of this study is to evaluate radiation dose reduction strategies in perfusion CT by using a biologic phantom. A formalin-preserved porcine liver was submerged in a 32-cm-wide acrylic phantom filled with water. The portal vein was connected to a continuous flow pump. The phantom was scanned with a perfusion CT protocol using 80 kVp and 400 mAs, every 1 second, for 50 seconds. This was repeated using 100 and 20 mAs. It was also repeated again using 400 mAs to assess reproducibility. A sparser scan frequency was simulated retrospectively. Blood flow was determined for each dataset using the maximum slope and deconvolution methods. Measurements of the mean blood flow values in identical regions of interest had a percent difference of 7% for repeated perfusion CT protocols using the same settings regardless of perfusion model used. The 100 mAs scans agreed with 400 mAs scans, with percent differences of 21% and 31% for the maximum slope and deconvolution methods, respectively. At a simulated frequency of one scan every 4 seconds, blood flow values differed up to 17% and 60% from the reference scan for the maximum slope and deconvolution methods, respectively. At 20 mAs and one scan every 1 second, or 400 mAs and a simulated frequency of one scan every 8 seconds, both computation methods failed to provide accurate blood flow estimates. The biologic phantom showed reproducible measurements that can help in optimizing perfusion CT protocols by determining both the acquisition parameters that affect radiation dose and the accuracy of estimates from different perfusion models.


The development of the ability of infants to utilize static cues to create and access representations of object shape.
A "transfer-across-depth-cues" method was used to explore the development of the ability to generate and use spatial representations of an object as specified by static pictorial depth cues. Infants were habituated to an object with depth specified by one cue and then presented with the same shape with depth specified by a different cue. Only if an abstract representation of that object had been formed could transfer across cues occur. Shading and line junctions uniquely determined the 3D shapes in these displays so that they appeared to be either a slice of cake with a flat top or a rocket. Without these cues, both line drawings were identical. Infants aged 6 to 7 months showed significant evidence of transfer, while infants aged 4 to 5 months did not. A control experiment demonstrated that the younger infants could discriminate between the objects when a single depth cue specified the shapes. These results are similar to our previous findings, which indicated that 6- to 7-month-old infants show transfer across shading and surface-contour cues, specifying convex and concave surfaces (A. Tsuruhara, T. Sawada, S. Kanazawa, M. K. Yamaguchi, & A. Yonas, 2009). This work supports the hypothesis that the ability to form 3D spatial representations from pictorial depth cues develops at about 6 months of age.


Barriers of Mucosal Entry of HIV/SIV.
Most new HIV infections, over 80%, occur through sexual transmission. During sexual transmission, the virus must bypass specific female and male reproductive tract anatomical barriers to encounter viable target cells. Understanding the generally efficient ability of these barrier to exclude HIV and the precise mechanisms of HIV translocation beyond these genital barriers is essential for vaccine and novel therapeutic development. In this review, we explore the mucosal, barriers of cervico-vaginal and penile tissues that comprise the female and male reproductive tracts. The unique cellular assemblies f the squamous and columnar epithelium are illustrate highlighting their structure and function. Each anatomical tissue offers a unique barrier to virus entry in healthy individuals. Unfortunately barrier dysfunction can lead to HIV transmission. How these diverse mucosal barriers have the potential to fail is considered, highlighting those anatomical areas that are postulated to offer a weaker barrier and are; therefore, more susceptible to viral ingress. Risk factors, such as sexually transmitted infections, microbiome dysbiosis, and high progestin environments are also associated with increased acquisition of HIV. How these states may affect the integrity of mucosal barriers leading to HIV acquisition are discussed suggesting mechanisms of transmission and revealing potential targets for intervention.


Identification of human GnIH homologs, RFRP-1 and RFRP-3, and the cognate receptor, GPR147 in the human hypothalamic pituitary axis.
The existence of a hypothalamic gonadotropin-inhibiting system has been elusive. A neuropeptide named gonadotropin-inhibitory hormone (GnIH, SIKPSAYLPLRF-NH(2)) which directly inhibits gonadotropin synthesis and release from the pituitary was recently identified in quail hypothalamus. Here we identify GnIH homologs in the human hypothalamus and characterize their distribution and biological activity. GnIH homologs were isolated from the human hypothalamus by immunoaffinity purification, and then identified as MPHSFANLPLRF-NH(2) (human RFRP-1) and VPNLPQRF-NH(2) (human RFRP-3) by mass spectrometry. Immunocytochemistry revealed GnIH-immunoreactive neuronal cell bodies in the dorsomedial region of the hypothalamus with axonal projections to GnRH neurons in the preoptic area as well as to the median eminence. RT-PCR and subsequent DNA sequencing of the PCR products identified human GnIH receptor (GPR147) mRNA expression in the hypothalamus as well as in the pituitary. In situ hybridization further identified the expression of GPR147 mRNA in luteinizing hormone producing cells (gonadotropes). Human RFRP-3 has recently been shown to be a potent inhibitor of gonadotropin secretion in cultured sheep pituitary cells by inhibiting Ca(2+) mobilization. It also directly modulates GnRH neuron firing. The identification of two forms of GnIH (RFRP-1 and RFRP-3) in the human hypothalamus which targets human GnRH neurons and gonadotropes and potently inhibit gonadotropin in sheep models provides a new paradigm for the regulation of hypothalamic-pituitary-gonadal axis in man and a novel means for manipulating reproductive functions.


Structural flexibility in proteins: impact of the crystal environment.
In the study of the structural flexibility of proteins, crystallographic Debye-Waller factors are the most important experimental information used in the calibration and validation of computational models, such as the very successful elastic network models (ENMs). However, these models are applied to single protein molecules, whereas the experiments are performed on crystals. Moreover, the energy scale in standard ENMs is undefined and must be obtained by fitting to the same data that the ENM is trying to predict, reducing the predictive power of the model. We develop an elastic network model for the whole protein crystal in order to study the influence of crystal packing and lattice vibrations on the thermal fluctuations of the atom positions. We use experimental values for the compressibility of the crystal to establish the energy scale of our model. We predict the elastic constants of the crystal and compare with experimental data. Our main findings are (1) crystal packing modifies the atomic fluctuations considerably and (2) thermal fluctuations are not the dominant contribution to crystallographic Debye-Waller factors. The programs developed for this work are available as supplementary material at Bioinformatics Online.


Incomplete insertion of pedicle screws in a standard construct reduces the fatigue life: A biomechanical analysis.
Pedicle screws are commonly used for posterior stabilization of the spine. When used in deformed or degenerated segments, the pedicle screws are often not fully inserted into the bone, but instead the threaded portion is exposed by 1 or 2 threads to accommodate rod placement and ensure alignment between the tulip of the screw and the rod. However, broken pedicle screws have been reported with the use of this method. The aim of this study was to determine how the fatigue life of the screw is affected by not fully inserting the screw into the bone. Spinal constructs were evaluated in accordance with ASTM F1717. The following three screw positions were subjected to compression bending fatigue loading; (i) pedicle screw fully inserted in the test block with no threads exposed (EXP-T0), (ii) pedicle screw inserted with one thread exposed outside the test block (EXP-T1), (iii) pedicle screw inserted with two threads exposed outside the test block (EXP-T2). Corresponding finite element models FEM-T0, FEM-T1 and FEM-T2 were also constructed and subjected to the same axial loading as the experimental groups to analyze the stress distribution in the pedicle screws and rods. The results showed that under a 190 N axial load, the EXP-T0 group survived the full 5 million cycles, the EXP-T1 group failed at 3.7 million cycles on average and the EXP-T2 groups failed at 1.0 million cycles on average, while the fatigue strength of both the EXP-T1 and EXP-T2 groups was 170 N. The constructs failed through fracture of the pedicle screw. In comparison to the FEM-T0 model, the maximum von Mises stress on the pedicle screw in the FEM-T1 and FEM-T2 models increased by 39% and 58%, respectively. In conclusion, this study demonstrated a drastic decrease in the fatigue life of pedicle screws when they are not full inserted into the plastic block.


CD4(+) T-lymphocytes mediate ischemia/reperfusion-induced inflammatory responses in mouse liver.
The success of orthotopic liver transplantation is dependent on multiple factors including MHC tissue compatibility and ischemic/reperfusion injury. Ischemic/reperfusion (I/R) injury in the liver occurs in a biphasic pattern consisting of both acute phase (oxygen free radical mediated) and subacute phase (neutrophil-mediated) damage. Although numerous studies have given insights into the process of neutrophil recruitment after I/R injury to the liver, the exact mechanism that initiates this subacute response remains undefined. Using a T cell-deficient mouse model, we present data that suggests that T-lymphocytes are key mediators of subacute neutrophil inflammatory responses in the liver after ischemia and reperfusion. To this end, using a partial lobar liver ischemia model, we compared the extent of reperfusion injury between immune competent BALB/c and athymic nu/nu mice. Studies evaluating the extent of liver damage as measured by serum transaminases (GPT) demonstrate similar acute (3-6 h) post-I/R responses in these two mouse models. In contrast, the subacute phase (16-20 h) of liver injury, as measured by both serum GPT levels and percent hepatocellular necrosis, was dramatically reduced in T cell-deficient mice as compared with those with an intact immune system. This reduction in liver injury seen in nu/nu mice was associated with a 10-fold reduction in hepatic neutrophil infiltration. Adoptive transfer of T cell-enriched splenocytes from immune competent mice was capable of reconstituting the neutrophil-mediated subacute inflammatory response within T cell-deficient nu/nu mice. Furthermore, in vivo antibody depletion of CD4(+) T-lymphocytes in immune competent mice resulted in a reduction of subacute phase injury and inflammation as measured by serum GPT levels and neutrophil infiltration. In contrast, depletion of CD8(+) T-lymphocytes had no effect on these indexes of subacute inflammation. Kinetic analysis of T cell infiltration in the livers of BALB/c mice demonstrated a fivefold increase in the number of hepatic CD4(+) T-lymphocytes within the first hour of reperfusion with no significant change in the number of CD8(+) T-lymphocytes. In summary, these results implicate CD4(+) T-lymphocytes as key regulators in initiating I/R-induced inflammatory responses in the liver. Such findings have implications for therapy directed at the early events in this inflammatory cascade that may prove useful in liver transplantation.


The changing structure of diets in the European Union in relation to healthy eating guidelines.
Our objective in this paper is to assess diets in the European Union (EU) in relation to the recommendations of the recent World Health Organization/Food and Agriculture Organization expert consultation and to show how diets have changed between 1961 and 2001. Computations make use of FAOSTAT data on food availability at country level linked to a food composition database to convert foods to nutrients. We further explore the growing similarity of diets in the EU by making use of a consumption similarity index. The index provides a single number measure of dietary overlap between countries. The data confirm the excessive consumption by almost all countries of saturated fats, cholesterol and sugars, and the convergence of nutrient intakes across the EU. Whereas in 1961 diets in several European countries were more similar to US diets than to those of other European countries, this is no longer the case; moreover, while EU diets have become more homogeneous, the EU as a whole and the USA have become less similar over time. Although the dominant cause of greater similarity in EU diets over the period studied is increased intakes in Mediterranean countries of saturated fats, cholesterol and sugar, also important are reductions in saturated fat and sugar in some Northern European countries. This suggests that healthy eating messages are finally having an impact on diets; a distinctly European diet may also be emerging.


Providing Higher Resolution Indicators of Rurality in the Surveillance, Epidemiology, and End Results (SEER) Database: Implications for Patient Privacy and Research.
The burden of cancer is higher in rural areas than urban areas. The NCI's Surveillance, Epidemiology, and End Results (SEER) database currently provides county-level information on rurality for cancer patients in its catchment area, but more nuanced measures of rurality would improve etiologic and surveillance studies. We analyzed disclosure risk and conducted a sample utility analysis of census tract-level measures of rurality, using (1) U.S. Department of Agriculture's Rural Urban Commuting Area (RUCA) codes and (2) U.S. Census data on percentage of the population living in nonurban areas. We evaluated the risk of disclosure by calculating the percentage of census tracts and cancer cases that would be uniquely identified by a combination of these two rurality measures with a census tract-level socioeconomic status (SES) variable. The utility analyses examined SES disparities across levels of rurality for lung and breast cancer incidence and relative survival. Risk of disclosure was quite low: <0.03% of census tracts and <0.03% of cancer cases were uniquely identified. Utility analyses demonstrated an SES gradient in lung and breast cancer incidence and survival, with relatively similar patterns across rurality variables. The RUCA and Census rurality measures have been added to a specialized SEER 18 database. Interested researchers can request access to this database to perform analyses of urban/rural differences in cancer incidence and survival. Such studies can provide important research support for future interventions to improve cancer prevention and control.


Appropriateness of antibiotic management of uncomplicated skin and soft tissue infections in hospitalized adult patients.
Skin and soft tissue infections (SSTIs) are a leading cause for hospitalizations in the United States. Few studies have addressed the appropriateness of antibiotic therapy in the management of SSTIs without complicating factors. We aimed to determine the appropriateness of antibiotic treatment duration for hospitalized adult patients with uncomplicated SSTIs. This was a retrospective analysis performed at two academic medical centers in Pittsburgh, Pennsylvania on patients aged 18 years and older with primary ICD-9 code for SSTIs admitted August 1st, 2014-March 31st, 2015. The primary outcome was the appropriateness of antibiotic treatment duration for uncomplicated SSTIs. Secondary objectives included the appropriateness of antibiotic agent spectrum, duration of inpatient length of stay (LOS), utilization of blood cultures and advanced imaging modalities, and re-hospitalization for SSTI within 30 days of discharge from the index admission. A total of 163 episodes were included in the cohort. The mean duration of total antibiotic therapy was 12.6 days. Appropriate duration was defined as receipt of total antibiotic duration of less than 10 days and occurred in 20.2% of patients. Twenty eight percent of patients received antibiotics for greater than 14 days. Seventy three (44.8%) patients received greater than 24 h of inappropriate extended spectrum gram-negative coverage; 65 (39.9%) received anaerobic coverage. In the majority of patients, treatment duration was excessive. Inappropriate broad spectrum antibiotic selection was utilized with regularity for SSTIs without complicating factors. The management of uncomplicated SSTIs represents a significant opportunity for antimicrobial stewardship.


An in vivo multimodal imaging study using MRI and PET of stem cell transplantation after myocardial infarction in rats.
The purpose of the study is to track iron-oxide nanoparticle-labelled adult rat bone marrow-derived stem cells (IO-rBMSCs) by magnetic resonance imaging (MRI) and determine their effect in host cardiac tissue using 2-deoxy-2-[F-18]fluoro-D: -glucose-positron emission tomography (FDG-PET). Infarcted rats were randomised to receive (1) live IO-rBMSCs by direct local injection, or (2) dead IO-rBMSCs as controls; (3) sham-operated rats received live IO-rBMSCs. The rats were then imaged from 2 days to 6 weeks post-cell implantation using both MRI at 9.4T and FDG-PET. Implanted IO-rBMSCs were visible in the heart by MRI for the duration of the study. Histological analysis confirmed that the implanted IO-rBMSCs were present for up to 6 weeks post-implantation. At 1 week post-IO-rBMSC transplantation, PET studies demonstrated an increase in FDG uptake in infarcted regions implanted with live IO-rBMSC compared to controls. Noninvasive multimodality imaging allowed us to visualise IO-rBMSCs and establish their affect on cardiac function in a rat model of myocardial infarction (MI).


Blood-brain barrier sodium transport limits development of brain edema during partial ischemia in gerbils.
Sodium derived from the blood is known to accumulate in brain tissue during the early stages of incomplete ischemia. Our present studies were undertaken to determine the relation between blood-brain barrier sodium transport and the development of ischemic brain edema. Incomplete cerebral ischemia was produced in gerbils by ligation of the left common carotid artery under ether anesthesia. Following recovery from the anesthetic, the gerbis were evaluated for the presence of neurologic symptoms and were divided into symptomatic (n = 77) and asymptomatic (n = 94) groups. Tissue water, sodium, and potassium contents, tissue plasma volume, and brain uptake of 22Na were measured in both groups 1.5, 3, 6, 12, and 24 hours after carotid ligation. There was a progressive accumulation of sodium and water in the ipsilateral cerebral cortex of the symptomatic group compared with either the corresponding contralateral cortex of the same gerbils or with the asymptomatic group. Net changes in brain sodium and potassium concentrations appeared to be the main determinants of fluid accumulation. Brain edema was not due to opening of the blood-brain barrier because the unidirectional transport of 22Na remained low and was even reduced by 35-55% in the ischemic cortex. Nevertheless, this sodium transport activity appeared to be rate-limiting in the development of brain edema during the first 3 hours of ischemia because the rate of sodium accumulation in the tissue was the same as the rate of 22Na transport from the blood to the brain. We conclude that blood-brain barrier sodium transport is an important factor in the formation of ischemic brain edema.


Analysis of Complications with Staged Surgery for Less Invasive Treatment of Adult Spinal Deformity.
Spinal deformity surgery is often invasive and lengthy. Staging surgery over separate operative days may reduce complications. Staging is often used in minimally invasive treatment of adult spinal deformity (ASD). To investigate the impact of staging on complication rates between hybrid (HYB; minimally invasive interbody with open posterior screw and rod fixation) and circumferential minimally invasive surgery (cMIS; minimally invasive interbody and screw/rod placement) procedures in patients with ASD. A multicenter database of patients with ASD was reviewed. Patients who underwent staging (at least 3 levels) and 2 years of follow-up were analyzed. A total of 99 patients underwent staging: 53 cMIS and 46 HYB surgeries. Propensity matching for levels fused resulted in 19 patients in each group. Intra- and perioperative complications were assessed. Three HYB but no cMIS intraoperative complications occurred. More HYB patients had perioperative complications than cMIS patients. Neurologic complications were more frequent in HYB versus cMIS. Other complications did not differ significantly. Thirty-day reoperations were higher with cMIS than HYB, but there was no difference in reoperation rate at long-term follow-up. cMIS patients had greater improvement in the Oswestry Disability Index. There was no difference in complications between staged versus unstaged cMIS surgeries. cMIS staged surgeries appear safer than HYB staged surgeries, and equally safe to cMIS unstaged surgeries. Perioperative complications were significantly higher for HYB staged surgeries. HYB surgeries may have better results when performed in a single setting, whereas cMIS surgeries can be performed in 1 or 2 stages depending on surgeon preference.


Aspergillus cell wall and biofilm.
The fungal cell is surrounded by a cell wall that acts as a sieve and a reservoir for effector molecules that play an active role during infection. This cell wall is essential for fungal growth as well as for resisting host defense mechanisms. The Aspergillus fumigatus cell wall is almost exclusively composed of polysaccharides. The fibrillar core is composed of a branched β-(1,3)-glucan to which chitin, β-(1,3)-/β-(1,4)-glucan, and galactomannan are covalently bound. The alkali-soluble amorphous fraction is mainly composed of α-(1,3)-glucan that has adhesive property and stabilizes the cell wall. Although the same polysaccharides are found in the cell wall of different A. fumigatus morphotypes (conidia and hyphae), their concentration and localization are different. Conidial (the morphotype that mainly enters host respiratory system) cell wall is covered by an outer layer of rodlets and melanin, which confers hydrophobic properties and imparts immunological inertness. In contrast, outer layer of the hypha contains galactosaminogalactan, recently identified as an A. fumigatus virulence factor. The hypha grows either as a network of agglutinated and hydrophobic mass (called mycelium) embedded in an extracellular matrix (ECM) rich in polysaccharides, hydrophobin, and melanin or segregated without ECM.


Identification of an Ire1alpha endonuclease specific inhibitor with cytotoxic activity against human multiple myeloma.
Activation of the adaptive Ire1-XBP1 pathway has been identified in many solid tumors and hematologic malignancies, including multiple myeloma (MM). Here, we report the identification of STF-083010, a novel small-molecule inhibitor of Ire1. STF-083010 inhibited Ire1 endonuclease activity, without affecting its kinase activity, after endoplasmic reticulum stress both in vitro and in vivo. Treatment with STF-083010 showed significant antimyeloma activity in model human MM xenografts. Similarly, STF-083010 was preferentially toxic to freshly isolated human CD138(+) MM cells compared with other similarly isolated cell populations. The identification of this novel Ire1 inhibitor supports the hypothesis that the Ire1-XBP1 axis is a promising target for anticancer therapy, especially in the context of MM.


A rare case of atlantooccipital dissociation in the context of occipitalization of the atlas, with a 2-year follow-up: case report.
Atlantooccipital dislocation (AOD) is a rare and often fatal injury. In cases of survival, residual deficits are severe and often include cranial nerve palsy, quadriplegia, or respiratory issues. Occipitalization is defined as partial or complete congenital fusion of the occiput to the atlas and is exceptionally rare. The authors present a rare case of AOD superimposed on a congenital occipitalization of the atlas. This 39-year-old man had AOD following a motor vehicle collision. On examination, his overall motor score on the American Spinal Injury Association scale was 5/100, and his rectal tone was absent. Computed tomography demonstrated AOD in an area of occipitalization. Magnetic resonance imaging revealed ligamentous injury leading to C1-2 instability. Intervention included occipital cervical instrumentation fusion from the occiput to C-3. Six months postoperatively, imaging revealed fusion of the graft and consolidation of the fractured occipitalization. At the 2-year follow-up, the patient's strength was 3/5 for wrist extension and handgrip on the right side and full strength in the rest of the myotomes. Bladder and bowel function was also normalized. A high-velocity collision led to disruption of the atlantooccipital ligaments and fracture of the occipitalized lateral masses in this patient. Internal fixation and fusion led to good fusion postoperatively. Occipitalization probably led to abnormal joint mechanics at the C1-occiput junction, which might have altered the amount of force required to fracture the occipitalization and produce AOD. This difference may partially account for the favorable neurological outcome in the featured patient compared with traditional cases of AOD.


Nucleus-aware Self-supervised Pretraining Using Unpaired Image-to-image Translation for Histopathology Images.
Self-supervised pretraining attempts to enhance model performance by obtaining effective features from unlabeled data, and has demonstrated its effectiveness in the field of histopathology images. Despite its success, few works concentrate on the extraction of nucleus-level information, which is essential for pathologic analysis. In this work, we propose a novel nucleus-aware self-supervised pretraining framework for histopathology images. The framework aims to capture the nuclear morphology and distribution information through unpaired image-to-image translation between histopathology images and pseudo mask images. The generation process is modulated by both conditional and stochastic style representations, ensuring the reality and diversity of the generated histopathology images for pretraining. Further, an instance segmentation guided strategy is employed to capture instance-level information. The experiments on 7 datasets show that the proposed pretraining method outperforms supervised ones on Kather classification, multiple instance learning, and 5 dense-prediction tasks with the transfer learning protocol, and yields superior results than other self-supervised approaches on 8 semi-supervised tasks. Our project is publicly available at https://github.com/zhiyuns/UNITPathSSL.


Comparing patient preferences and healthcare provider recommendations with the pen versus vial-and-syringe insulin delivery in patients with type 2 diabetes.
This study aimed to examine healthcare provider (HCP) recommendations and patient preferences for the insulin pen versus vial-and-syringe in patients with type 2 diabetes mellitus (T2DM) and to assess clinical end points and safety outcomes. Using a randomized, open-label, crossover design, in total, 405 insulin-naive adults with T2DM from 60 centers received basal insulin glargine in one of two device treatment sequences (2 weeks of pen followed by 2 weeks of vial-and-syringe, or vice versa). The primary end point, patient device preference, was evaluated at Week 4 (end of the crossover period) using the Insulin Injection Preference Questionnaire. Patient preference and HCP recommendation were assessed with one global item and three subscale items (blood glucose control, reluctance to use insulin, and long-term insulin use) using a 5-point scale ranging from 1=not preferred or not recommended to 5=preferred or recommended. Patients were then re-randomized to either pen or vial-and-syringe for further observation (6, 10, and 30 weeks) to evaluate clinical end points (glycosylated hemoglobin [A1C] and fasting blood glucose levels) and safety outcomes (hypoglycemia and adverse events). Patients reported a significant preference for pens over vial-and-syringe, and HCPs strongly recommended pens over vial-and-syringe (both P<0.001). Consistent response patterns were observed by HCPs and patients for the three subscale items. Fasting blood glucose, A1C levels, and the incidence of hypoglycemia were comparable in the two groups. Patients preferred pens over vial-and-syringe, with the pen device also recommended by HCPs, when initiating basal insulin treatment in insulin-naive patients with T2DM.


Non-genetic origins of cell-to-cell variability in TRAIL-induced apoptosis.
In microorganisms, noise in gene expression gives rise to cell-to-cell variability in protein concentrations. In mammalian cells, protein levels also vary and individual cells differ widely in their responsiveness to uniform physiological stimuli. In the case of apoptosis mediated by TRAIL (tumour necrosis factor (TNF)-related apoptosis-inducing ligand) it is common for some cells in a clonal population to die while others survive-a striking divergence in cell fate. Among cells that die, the time between TRAIL exposure and caspase activation is highly variable. Here we image sister cells expressing reporters of caspase activation and mitochondrial outer membrane permeabilization after exposure to TRAIL. We show that naturally occurring differences in the levels or states of proteins regulating receptor-mediated apoptosis are the primary causes of cell-to-cell variability in the timing and probability of death in human cell lines. Protein state is transmitted from mother to daughter, giving rise to transient heritability in fate, but protein synthesis promotes rapid divergence so that sister cells soon become no more similar to each other than pairs of cells chosen at random. Our results have implications for understanding 'fractional killing' of tumour cells after exposure to chemotherapy, and for variability in mammalian signal transduction in general.


Comparison of subjective and objective methods to determine the retinal arterio-venous ratio using fundus photography.
To assess the inter and intra observer variability of subjective grading of the retinal arterio-venous ratio (AVR) using a visual grading and to compare the subjectively derived grades to an objective method using a semi-automated computer program. Following intraocular pressure and blood pressure measurements all subjects underwent dilated fundus photography. 86 monochromatic retinal images with the optic nerve head centred (52 healthy volunteers) were obtained using a Zeiss FF450+ fundus camera. Arterio-venous ratios (AVR), central retinal artery equivalent (CRAE) and central retinal vein equivalent (CRVE) were calculated on three separate occasions by one single observer semi-automatically using the software VesselMap (ImedosSystems, Jena, Germany). Following the automated grading, three examiners graded the AVR visually on three separate occasions in order to assess their agreement. Reproducibility of the semi-automatic parameters was excellent (ICCs: 0.97 (CRAE); 0.985 (CRVE) and 0.952 (AVR)). However, visual grading of AVR showed inter grader differences as well as discrepancies between subjectively derived and objectively calculated AVR (all p<0.000001). Grader education and experience leads to inter-grader differences but more importantly, subjective grading is not capable to pick up subtle differences across healthy individuals and does not represent true AVR when compared with an objective assessment method. Technology advancements mean we no longer rely on opthalmoscopic evaluation but can capture and store fundus images with retinal cameras, enabling us to measure vessel calibre more accurately compared to visual estimation; hence it should be integrated in optometric practise for improved accuracy and reliability of clinical assessments of retinal vessel calibres.


Visualizing pair formation on the atomic scale and the search for the mechanism of superconductivity in high-T(c) cuprates.
We have developed several new experimental techniques, based on the scanning tunneling microscope, to visualize the process of pair formation on the atomic scale and to probe with high precision what controls the strength of pairing in high-T(c) cuprate superconductor compounds. These new experiments provide evidence that pairing in these exotic superconductors occurs above the bulk transition temperature and in nanoscale regions with sizes of 1-3 nm. The high temperature nucleation and proliferation of these nanoscale puddles have a strong connection to the temperature-doping phase diagram of these superconductors. On average we have found that the pairing gap Δ and the temperature at which they first nucleate T(p) follow the simple relation: 2Δ/k(B)T(p)∼8. Moreover, the variations of the pairing strength on the nanoscale can be examined to find microscopic clues to the mechanism of pairing. Specifically, we have found evidence that suggests that strong electronic correlation, as opposed to coupling of electrons to bosons, is responsible for the pairing mechanism in the cuprates. Surprisingly, we have found that nanoscale measurements of electronic correlations in the normal state (at temperatures as high as twice T(c)) can be used to predict the strength of the local pairing interaction at low temperatures.


IL-12-and IL-23 in health and disease.
Interleukin (IL)-12 and IL-23 play important roles in the development of experimental autoimmune disease models and numerous afflictions affecting humans. Preclinical data over the last 20 years combined with successful clinical trials has identified a clear relationship between IL-12, IL-23 and the generation of pathogenic T helper cells capable of orchestrating tissue inflammation. Observations made in the clinic have shown that IL-12p40, a common subunit shared by IL-12 and IL-23, is critical to pathologies associated with psoriasis, inflammatory bowel disease (IBD) and tumor growth. These advancements have set in motion the development of a number of potential therapeutics aimed at manipulating IL-12/23 signaling pathways in both mice and humans. This review will discuss a brief history of the understanding and expansion of the IL-12 cytokine family, some difficulties associated with preclinical data interpretation and finally the medicinal interventions that have been developed to combat IL-12/23-driven autoimmune disorders.


Comparative study of transdermal drug delivery systems of resveratrol: High efficiency of deformable liposomes.
Trans-resveratrol (3, 5, 4' trihydroxystilbene, RSV) is a natural compound that shows antioxidant, cardioprotective, anti-inflammatory and anticancer properties. The transdermal, painless application of RSV is an attractive option to other administration routes owing to its several advantages like avoiding gastrointestinal problems and first pass metabolism. However, its therapeutic potential is limited by its low solubility and low stability in water and the reduced permeability of stratum corneum. To overcome these inconveniences the encapsulation of this compound in a drug delivery system is proposed here. In order to find the best carrier for transdermal application of RSV various liposomal nanoparticulate carriers like conventional liposomes (L-RSV), deformable liposomes (LD-RSV), ultradeformable liposomes (LUD-RSV) and ethosomes (Etho-RSV) were assayed. Transmission electron microscopic (TEM) and dynamic light scattering (DLS) studies were performed to analyze the surface morphology of these carriers. Structural characterization for these formulations was performed by confocal Raman spectroscopy. The spectroscopic results were analysed in conjunction with calorimetric data to identify the conformational changes and stability of formulations in the different nanoparticles induced by the presence of RSV. Comparison of the results obtained with the different carrier systems (L-RSV, LD-RSV, LUD-RSV and Etho-RSV) revealed that the best RSV carrier was LD-RSV. The increase in the fluidity of the bilayers in the region of the hydrophobic chains of the phospholipid by ethanol probably facilitates the accommodation of the RSV in the bilayer and contributes to the improved encapsulation of RSV without affecting the mobility of this carrier.


Membrane-type 1 matrix metalloproteinase cleaves CD44 and promotes cell migration.
Migratory cells including invasive tumor cells frequently express CD44, a major receptor for hyaluronan and membrane-type 1 matrix metalloproteinase (MT1-MMP) that degrades extracellular matrix at the pericellular region. In this study, we demonstrate that MT1-MMP acts as a processing enzyme for CD44H, releasing it into the medium as a soluble 70-kD fragment. Furthermore, this processing event stimulates cell motility; however, expression of either CD44H or MT1-MMP alone did not stimulate cell motility. Coexpression of MT1-MMP and mutant CD44H lacking the MT1-MMP-processing site did not result in shedding and did not promote cell migration, suggesting that the processing of CD44H by MT1-MMP is critical in the migratory stimulation. Moreover, expression of the mutant CD44H inhibited the cell migration promoted by CD44H and MT1-MMP in a dominant-negative manner. The pancreatic tumor cell line, MIA PaCa-2, was found to shed the 70-kD CD44H fragment in a MT1-MMP-dependent manner. Expression of the mutant CD44H in the cells as well as MMP inhibitor treatment effectively inhibited the migration, suggesting that MIA PaCa-2 cells indeed use the CD44H and MT1-MMP as migratory devices. These findings revealed a novel interaction of the two molecules that have each been implicated in tumor cell migration and invasion.