Single-Nucleotide Polymorphisms Associated With Age-Related Macular Degeneration and Lesion Phenotypes in the Comparison of Age-Related Macular Degeneration Treatments Trials.
Single-nucleotide polymorphisms (SNPs) associated with the CFH, ARMS2, C3, LIPC, CFB, and C2 genes are associated with age-related macular degeneration (AMD); however, the association of these SNPs with angiographic features of neovascular AMD has been inconsistent in previous studies, and to date, no studies have addressed their association with features on optical coherence tomography. To evaluate the influence of genotype of SNPs previously associated with AMD on the phenotype of neovascular lesions. Participants for this cross-sectional study were recruited from the 1185 patients enrolled in the Comparison of Age-Related Macular Degeneration Treatments Trials (CATT), a randomized clinical trial. Eligibility criteria for CATT specified that eyes have choroidal neovascularization and visual acuity between 20/25 and 20/320. A subgroup of 835 patients provided blood samples from July 2010 through September 2011 and were genotyped for the SNPs rs1061170 (CFH), rs10490924 (ARMS2),rs2230199 (C3), rs10468017 (LIPC), rs4151667 (CFB), rs547154 (C2) using TaqMan SNP genotyping assays. Data analysis was initiated in November 2013 and completed in January 2016. Pretreatment ocular characteristics on fluorescein angiography (lesion type, area of neovascularization and total lesion, retinal angiomatous proliferation) and on time-domain optical coherence tomography (presence of intraretinal, subretinal, and subretinal pigment epithelium fluid; thickness at the foveal center of the retina, subretinal fluid, and subretinal tissue complex), visual acuity, and age. A total of 835 (73%) of 1150 CATT patients were genotyped. Mean age decreased with the number of risk alleles for CFH (P < .001), ARMS2 (P < .001), and C3 (P = .005). The following results were found as the number of risk alleles increased from 0 to 1 to 2. For CFH, mean total thickness decreased from 476 to 476 to 434 µm (P = .01; adjusted for age, sex, and smoking status). For ARMS2, the mean area of the total lesion increased from 2.0 to 2.8 to 2.4 mm2 (P = .03), the proportion with retinal angiomatous proliferation lesions increased from 8% to 10% to 12% (P = .05), and the proportion with intraretinal fluid increased from 72% to 71% to 82% (P = .008). For C3, the proportion with intraretinal fluid decreased from 78% to 69% to 64% (P = .001), and the mean retinal thickness decreased from 225 to 207 to 197 µm (P = .02). CFH, ARMS2, and C3 were associated with specific features of neovascularization at the time patients were enrolled in CATT. Previously identified associations of ARMS2 and CFH with type of choroidal neovascularization on fluorescein angiography were not confirmed. New associations with OCT features identified in CATT need confirmation to establish whether a true association exists. clinicaltrials.gov Identifier: NCT00593450.


A slit method to determine the focal spot size and shape of TomoTherapy system.
To obtain accurate x-ray source profile measurements using a slit-collimator, the slit-collimator should have a narrow width, large height, and be positioned near the source. However, these conditions may not always be met. In this paper, the authors provide a detailed analysis of the slit measurement geometry and the relationship between the slit parameters and the measured x-ray source profile. The slit model allows the use of a shorter and more easily available slit-collimator, while accurate source profile measurements can still be obtained. Measurements were performed with a variety of slit widths and/or slit to source distances. The relationship derived between the slit parameters and the measured profile was used to determine the true focal spot profile through a least square fit of the profile data. The model was verified by comparing the predicted profiles at a variety of slit-collimator parameters with the measured results on the TomoTherapy Hi-Art system. Both the treatment beam and the imaging beam were measured. For treatment mode, it was found that a source consisting of one Gaussian with a 0.75 mm full-width-half-maximum (FWHM) and 72% peak amplitude and a second Gaussian with a 2.27 mm FWHM and 18% peak amplitude matched measurement profiles. The overall source profile has a FWHM of 0.93 mm, but with a higher amplitude in the tail region than a single Gaussian. For imaging mode, the source consists of one Gaussian with a 0.68 mm FWHM and 82% peak amplitude and a second Gaussian with a 1.83 mm FWHM and 18% peak amplitude. The overall source profile has a FWHM of 0.77 mm. Our study of the focal spot measurement using slit-collimators showed that accurate source profile measurements can be achieved through fitting of measurement results at different slit widths and source-to-slit distances (SSD). Quantitative measurements of the TomoTherapy linac focal spot showed that the source distribution could be better described with a model consisting of two Gaussian components rather than a single Gaussian model as assumed in previous studies.


Detecting Flying Objects Using a Single Moving Camera.
We propose an approach for detecting flying objects such as Unmanned Aerial Vehicles (UAVs) and aircrafts when they occupy a small portion of the field of view, possibly moving against complex backgrounds, and are filmed by a camera that itself moves. We argue that solving such a difficult problem requires combining both appearance and motion cues. To this end we propose a regression-based approach for object-centric motion stabilization of image patches that allows us to achieve effective classification on spatio-temporal image cubes and outperform state-of-the-art techniques. As this problem has not yet been extensively studied, no test datasets are publicly available. We therefore built our own, both for UAVs and aircrafts, and will make them publicly available so they can be used to benchmark future flying object detection and collision avoidance algorithms.


Dignity in the terminally ill: a cross-sectional, cohort study.
Considerations of dignity are often raised in reference to the care of dying patients. However, little research that addresses this issue has been done. Our aim was to identify the extent to which dying patients perceive they are able to maintain a sense of dignity, and to ascertain how demographic and disease-specific variables relate to the issue of dignity in these individuals. We did a cross-sectional study of a cohort of terminally ill patients with cancer, who had a life expectancy of less than 6 months. We enrolled 213 patients from two palliative care units in Winnipeg, Canada, and asked them to rate their sense of dignity. Our main outcome measures included: a 7-point sense of dignity item; the symptom distress scale; the McGill pain questionnaire; the index of independence in activities of daily living (IADL); a quality of life scale; a brief battery of self-report measures, including screening for desire for death, anxiety, hopelessness, and will to live; burden to others; and requirement for social support. 16 of 213 patients (7.5%; 95% CI 4-11) indicated that loss of dignity was a great concern. These patients were far more than likely than the rest of the cohort to report psychological distress and symptom distress, heightened dependency needs, and loss of will to live. Loss of dignity is closely associated with certain types of distress often seen among the terminally ill. Preservation of dignity should be an overall aim of treatment and care in patients who are nearing death.


Estrogen and progestin components of oral contraceptives: relationship to vascular disease.
Recently, new information has been published about: a) the relationship between combination oral contraceptives (OCs), estrogen dose, cigarette smoking, and the risk of myocardial infarction (MI) and stroke; and b) the effect of different progestins on the risk of venous thromboembolism (VTE). We review the epidemiologic data. Regardless of age, in the absence of smoking, use of sub-50 micrograms OCs is not associated with any meaningful increase in risk of MI or stroke. If the small, statistically nonsignificant elevations in risk for these diseases are assumed (for the sake of argument) to be causal, then the incidence of MI and stroke associated with use of OCs containing less than 50 micrograms ethinyl estradiol (EE) would be approximately 2 per 100,000 per year. For women less than 35 years of age who do not smoke or do not have a history of hypertension, the risk would be even lower. Any woman over the age of 35 who smokes should be advised to use a non-estrogen or nonhormonal contraceptive. There are now two reports, from jick et al. and Lewis et al., that demonstrate that the relative risk of MI is certainly no greater for users of OCs containing desogestrel or gestodene than for users of OCs containing older progestins. In fact, both show reduced relative risks for the newer progestins compared to the older ones. With respect to progestins, four recent epidemiologic studies have indicated a twofold increased risk of nonfatal VTE with use of OCs containing desogestrel or gestodene compared with levonorgestrel. A fifth report, which showed an increased relative risk for norgestimate, is based on use among only 19 cases and 31 controls and is not statistically significant. As the authors themselves caution and as subsequent follow-up analyses and editorials conclude, these studies do not provide evidence for a cause-and-effect relationship between OCs containing desogestrel or gestodene, and VTE. The recommendation with respect to desogestrel- and gestodene-containing OCs is that no change in prescribing practices is warranted for either current or new-start patients. There is a growing body of evidence demonstrating that OCs containing 30 or 35 micrograms of EE have lower risks of MI, stroke, and VTE than higher dose OCs. However, there is no epidemiologic study that demonstrates a greater risk of vascular events among women using OCs containing 30 or 35 micrograms EE compared with preparations containing 20 micrograms EE. Users of sub-50 micrograms OCs of any age have no clinically meaningful increase in incidence of MI or stroke compared with non-OC users. This is also true for smokers under the age of 35 years who use OCs. However, smokers over the age of 35 years who use OCs still have an unacceptably high incidence rate of MI and stroke and should not use combination OCs. Sub-50 micrograms OCs of all types are associated with a small excess risk of VTE, about 15 per 100,000 events per year. Until there is biologic explanation of the twofold greater risk of VTE in users of OCs containing desogestrel or gestodene compared with users of those containing older progestins, this association should not be accepted as one of cause and effect.


Quality and effectiveness of different approaches to primary care delivery in Brazil.
Since 1994, Brazil has developed a primary care system based on multidisciplinary teams which include not only a physician and a nurse, but also 4-6 lay community health workers. This system now consists of 26,650 teams, covering 46% of the Brazilian population. Yet relatively few investigations have examined its effectiveness, especially in contrast with that of the traditional multi-specialty physician team approach it is replacing, or that of other existing family medicine approaches placing less emphasis on lay community health workers. Primary health care can be defined through its domains of access to first contact, continuity, coordination, comprehensiveness, community orientation and family orientation. These attributes can be ascertained via instruments such as the Primary Care Assessment Tool (PCATool), and correlated with the effectiveness of care. The objectives of our study are to validate the adult version of this instrument in Portuguese, identify the extent (quality) of primary care present in different models of primary care services, and correlate this extent with measures of process and outcomes in patients with diabetes, hypertension and coronary heart disease (CHD). We are conducting a population-based cross-sectional study of primary care in the municipality of Porto Alegre. We will interview a random sample totaling 3000 adults residing in geographic areas covered by four distinct models of primary care of the Brazilian national health system or, alternatively, by one nationally prominent complementary health care service, as well as the physicians and nurses of the health teams of these services. Interviews query perceived quality of care (PCATool-Adult Version), patient satisfaction, and process indicators of management of diabetes, hypertension and known CHD. We are measuring blood pressure, anthropometrics and, in adults with known diabetes, glycated hemoglobin. We hope to contribute not only by validating the PCATool-Adult Version for use in Brazil, but also by furnishing ample data concerning the appropriate mix of health care professionals in the primary care team, a question of international import. Once validated, future use of this instrument should help direct advances aiming at improving the quality of primary care in Brazil.


Docking, triggering, and biological activity of dynemicin A in DNA: a computational study.
The triggering and biological activity of the naturally occurring enediyne dynemicin A (1) was investigated, both inside and outside the minor groove of the duplex 10-mer B-DNA sequence d(CTACTACTGG).d(CCAGTAGTAG), using density functional theory (B3LYP with the 3-21G and 6-31G(d) basis set), BD(T)/cc-pVDZ (Brueckner doubles with a perturbative treatment of triple excitations), and the ONIOM approach. Enediyne 1 is triggered by NADPH in a strongly exothermic reaction (-88 kcal/mol), which involves a number of intermediate steps. Untriggered 1 has a high barrier for the Bergman cyclization (52 kcal/mol) that is lowered after triggering to 16.7 kcal/mol due to an epoxide opening and the accompanying strain relief. The Bergman reaction of triggered 1 is slightly exothermic by 2.8 kcal/mol. The singlet biradical formed in this reaction is kinetically stable (activation enthalpies of 19.5 and 21.8 kcal/mol for retro-Bergman reactions) and is as reactive as para-benzyne. The activity-relevant docking mode is an edge-on insertion into the minor groove, whereas the intercalation between base pairs, although leading to larger binding energies, excludes a triggering of 1 and the development of its biological activity. Therefore, an insertion-intercalation model is developed, which can explain all known experimental observations made for 1. On the basis of the insertion-intercalation model it is explained why large intercalation energies suppress the biological activity of dynemicin and why double-strand scission can be achieved only in a two-step mechanism that involves two enediyne molecules, explaining thus the high ratio of single-strand to double-strand scission observed for 1.


Silica-based nano-coating on zirconia surfaces using reactive magnetron sputtering: effect on chemical adhesion of resin cements.
To compare the effect of silica (Si)-based nano-coating deposited by reactive magnetron sputtering (RMP) with that of conventional surface conditioning using metal/zirconia primer alone or after air-particle abrasion on the adhesion of resin cements to zirconia ceramic. Two hundred forty zirconia ceramic blocks (Cercon) were sintered, finished with 1200- grit SiC paper under water cooling, and cleaned ultrasonically in distilled water for 10 min. The blocks (4.5 mm x 3.5 mm x 4.5 mm) were randomly divided into 24 groups (n = 10) according to 3 testing parameters: a) resin cements (Multilink, Panavia F, RelyX U100), b) surface conditioning (no conditioning as control group; Metal/Zirconia Primer; air abrasion + Metal/Zirconia Primer; Si-based nanofilm + Monobond s); c) aging (no aging vs thermocycling at 5°C to 55°C, 6000 cycles). The nanofilm was deposited by direct current using argon/oxygen plasma (8:1 in flux) on the zirconia surface. Resin cements were bonded to zirconia surfaces using polyethylene molds. The shear bond strength (SBS) test was performed using a universal testing machine (1 mm/min), and after debonding, the substrate and adherent surfaces were analyzed using optical and scanning electron microscopes to categorize the failure types. The data were statistically evaluated using 3-way ANOVA and Tukey's test (5%). Resin cement type (p < 0.05), surface conditioning method (p < 0.05), and aging condition (p < 0.05) had a significant effect on the bond strength results. Interactions were also significant (p < 0.05). In the nonaged condition, while control groups presented the lowest results with all cements (0 to 5.2 MPa), the airabraded group in combination with RelyX U100 resulted in the highest SBS (21.8 ± 6.7 MPa). After aging, the SBS results decreased in the air-abraded groups for all cements (4.54 to 9.44 MPa) and showed no statistical significance compared to the Si-based nanocoated groups (4.24 to 6.44 MPa). After air-abrasion and primer application, only Panavia F and RelyX U100 cements showed exclusively mixed failures, but after nanofilm coating and silanization, all cements showed exclusively mixed failures with and without aging. Chemical adhesion of the resin cements tested to zirconia was similar after silica-based nanofilm deposition and air abrasion followed by primer application.


Evaluation of imatinib mesylate in the treatment of pulmonary arterial hypertension.
Imatinib mesylate is a small molecule inhibitor that selectively inhibits the PDGF receptor kinase as well the cKIT and Abl kinases, among other targets. Various studies have implicated the PDGF pathway in the pathogenesis of pulmonary arterial hypertension (PAH). Inhibition with imatinib mesylate has shown efficacy in human case reports and experimental models of PAH. Results from a Phase II trial of imatinib mesylate in PAH did not meet the primary end point but showed improvement in several secondary end points and in a subgroup analysis. As suggested by this study as well as a few case reports, imatinib may be effective in a subset of patients with more severe disease. However, this remains to be further validated through a Phase III study, which is already underway. In conclusion, it appears that imatinib mesylate may hold promise as an adjunct drug in PAH therapy, especially since it is directed at a pathway not previously targeted.


Behavioural insights and the evolving COVID-19 pandemic.
Behavioural sciences have complemented medical and epidemiological sciences in the response to the SARS-CoV-2 pandemic. As vaccination uptake continues to increase across the EU/EEA - including booster vaccinations - behavioural science research remains important for both pandemic policy, planning of services and communication. From a behavioural perspective, the following three areas are key as the pandemic progresses: (i) attaining and maintaining high levels of vaccination including booster doses across all groups in society, including socially vulnerable populations, (ii) informing sustainable pandemic policies and ensuring adherence to basic prevention measures to protect the most vulnerable population, and (iii) facilitating population preparedness and willingness to support and adhere to the reimposition of restrictions locally or regionally whenever outbreaks may occur. Based on mixed-methods research, expert consultations, and engagement with communities, behavioural data and interventions can thus be important to prevent and effectively respond to local or regional outbreaks, and to minimise socioeconomic and health disparities. In this Perspective, we briefly outline these topics from a European viewpoint, while recognising the importance of considering the specific context in individual countries.


Hypomethylation signature of tumor-initiating cells predicts poor prognosis of ovarian cancer patients.
DNA methylation contributes to tumor formation, development and metastasis. Epigenetic dysregulation of stem cells is thought to predispose to malignant development. The clinical significance of DNA methylation in ovarian tumor-initiating cells (OTICs) remains unexplored. We analyzed the methylomic profiles of OTICs (CP70sps) and their derived progeny using a human methylation array. qRT-PCR, quantitative methylation-specific PCR (qMSP) and pyrosequencing were used to verify gene expression and DNA methylation in cancer cell lines. The methylation status of genes was validated quantitatively in cancer tissues and correlated with clinicopathological factors. ATG4A and HIST1H2BN were hypomethylated in OTICs. Methylation analysis of ATG4A and HIST1H2BN by qMSP in 168 tissue samples from patients with ovarian cancer showed that HIST1H2BN methylation was a significant and independent predictor of progression-free survival (PFS) and overall survival (OS). Multivariate Cox regression analysis showed that patients with a low level of HIST1H2BN methylation had poor PFS (hazard ratio (HR), 4.5; 95% confidence interval (CI), 1.4-14.8) and OS (HR, 4.3; 95% CI, 1.3-14.0). Hypomethylation of both ATG4A and HIST1H2BN predicted a poor PFS (HR, 1.8; 95% CI, 1.0-3.6; median, 21 months) and OS (HR, 1.7; 95% CI, 1.0-3.0; median, 40 months). In an independent cohort of ovarian tumors, hypomethylation predicted early disease recurrence (HR, 1.7; 95% CI, 1.1-2.5) and death (HR, 1.4; 95% CI, 1.0-1.9). The demonstration that expression of ATG4A in cells increased their stem properties provided an indication of its biological function. Hypomethylation of ATG4A and HIST1H2BN in OTICs predicts a poor prognosis for ovarian cancer patients.


Effectiveness of Deep Water Running on Improving Cardiorespiratory Fitness, Physical Function and Quality of Life: A Systematic Review.
Deep Water Running (DWR) is a form of aquatic aerobic exercise simulating the running patterns adopted on dry land. Little is known on the effectiveness of DWR despite gaining popularity. The objective of this study is to systematically review the effects of DWR on cardiorespiratory fitness, physical function, and quality of life in healthy and clinical populations. A systematic search was completed using six databases, including SPORTDiscus, MEDLINE, CINAHL, AMED, Embase, and The Cochrane Library, up to February 2022. Eleven studies evaluating the effectiveness of DWR on cardiorespiratory fitness (CRF), physical function, or quality of life (QoL), compared with no interventions (or standard treatment) or land-based trainings were identified. Data relevant to the review questions were extracted by two independent reviewers when means and standard deviations were reported, and standardized mean differences were calculated. A quality assessment was conducted using selected items from the Downs and Black checklist. A total of 11 clinical trials (7 randomized controlled trials) with a total of 287 participants were included. Meta-analyses were not completed due to the high level of clinical and statistical heterogeneity between studies. Compared with land-based training, DWR showed similar effects on CRF with limited studies reporting outcomes of physical function and QoL compared with the no-exercise control group. DWR appears to be comparable to land-based training for improving CRF. The aquatic environment may provide some advantages for off-loaded exercise at high intensity in populations that are weak, injured or in pain, but more studies are required.


"Latent" infection with Toxoplasma gondii: association with trait aggression and impulsivity in healthy adults.
Latent chronic infection with Toxoplasma gondii (T. gondii), a common neurotropic pathogen, has been previously linked with suicidal self-directed violence (SSDV). We sought to determine if latent infection with T. gondii is associated with trait aggression and impulsivity, intermediate phenotypes for suicidal behavior, in psychiatrically healthy adults. Traits of aggression and impulsivity were analyzed in relationship to IgG antibody seropositivity for T. gondii and two other latent neurotropic infections, herpes simplex virus 1 (HSV1) and cytomegalovirus (CMV). One thousand community-residing adults residing in the Munich metropolitan area with no Axis I or II conditions by SCID for DSM-IV (510 men, 490 women, mean age 53.6 ± 15.8, range 20-74). Plasma samples were tested for IgG antibodies to T. gondii, HSV-1 and CMV by ELISA. Self-reported ratings of trait aggression scores (Questionnaire for Measuring Factors of Aggression [FAF]) and trait impulsivity (Sensation-Seeking Scale-V [SSS-V]) were analyzed using linear multivariate methods. T. gondii IgG seropositivity was significantly associated with higher trait reactive aggression scores among women (p < .01), but not among men. T. gondii-positivity was also associated with higher impulsive sensation-seeking (SSS-V Disinhibition) among younger men (p < .01) aged 20-59 years old (median age = 60). All associations with HSV-1 and CMV were not significant. Aggression and impulsivity, personality traits considered as endophenotypes for SSDV, are associated with latent T. gondii infection in a gender and age-specific manner, and could be further investigated as prognostic and treatment targets in T. gondii-positive individuals at risk for SSDV.


Diet-dependent gene expression in honey bees: honey vs. sucrose or high fructose corn syrup.
Severe declines in honey bee populations have made it imperative to understand key factors impacting honey bee health. Of major concern is nutrition, as malnutrition in honey bees is associated with immune system impairment and increased pesticide susceptibility. Beekeepers often feed high fructose corn syrup (HFCS) or sucrose after harvesting honey or during periods of nectar dearth. We report that, relative to honey, chronic feeding of either of these two alternative carbohydrate sources elicited hundreds of differences in gene expression in the fat body, a peripheral nutrient-sensing tissue analogous to vertebrate liver and adipose tissues. These expression differences included genes involved in protein metabolism and oxidation-reduction, including some involved in tyrosine and phenylalanine metabolism. Differences between HFCS and sucrose diets were much more subtle and included a few genes involved in carbohydrate and lipid metabolism. Our results suggest that bees receive nutritional components from honey that are not provided by alternative food sources widely used in apiculture.


Genomic profiling reveals distinctive molecular relapse patterns in IDH1/2 wild-type glioblastoma.
Molecular changes associated with the progression of glioblastoma after standard radiochemotherapy remain poorly understood. We compared genomic profiles of 27 paired primary and recurrent IDH1/2 wild-type glioblastomas by genome-wide array-based comparative genomic hybridization. By bioinformatic analysis, primary and recurrent tumor profiles were normalized and segmented, chromosomal gains and losses identified taking the tumor cell content into account, and difference profiles deduced. Seven of 27 (26%) pairs lacked DNA copy number differences between primary and recurrent tumors (equal pairs). The recurrent tumors in 9/27 (33%) pairs contained all chromosomal imbalances of the primary tumors plus additional ones, suggesting a sequential acquisition of and/or selection for aberrations during progression (sequential pairs). In 11/27 (41%) pairs, the profiles of primary and recurrent tumors were divergent, i.e., the recurrent tumors contained additional aberrations but had lost others, suggesting a polyclonal composition of the primary tumors and considerable clonal evolution (discrepant pairs). Losses on 9p21.3 harboring the CDKN2A/B locus were significantly more common in primary tumors from sequential and discrepant (nonequal) pairs. Nonequal pairs showed ten regions of recurrent genomic differences between primary and recurrent tumors harboring 46 candidate genes associated with tumor recurrence. In particular, copy numbers of genes encoding apoptosis regulators were frequently changed at progression. In summary, approximately 25% of IDH1/2 wild-type glioblastoma pairs have stable genomic imbalances. In contrast, approximately 75% of IDH1/2 wild-type glioblastomas undergo further genomic aberrations and alter their clonal composition upon recurrence impacting their genomic profile, a process possibly facilitated by 9p21.3 loss in the primary tumor. © 2014 Wiley Periodicals, Inc.


Quantitative assessment of barriers to the clinical development and adoption of cellular therapies: A pilot study.
There has been a large increase in basic science activity in cell therapy and a growing portfolio of cell therapy trials. However, the number of industry products available for widespread clinical use does not match this magnitude of activity. We hypothesize that the paucity of engagement with the clinical community is a key contributor to the lack of commercially successful cell therapy products. To investigate this, we launched a pilot study to survey clinicians from five specialities and to determine what they believe to be the most significant barriers to cellular therapy clinical development and adoption. Our study shows that the main concerns among this group are cost-effectiveness, efficacy, reimbursement, and regulation. Addressing these concerns can best be achieved by ensuring that future clinical trials are conducted to adequately answer the questions of both regulators and the broader clinical community.


Association of Urinary Levels of Bisphenols A, F, and S with Endometriosis Risk: Preliminary Results of the EndEA Study.
Aim: The aim of this study was to explore associations of urinary concentrations of bisphenols A (BPA), S (BPS), and F (BPF) and of thiobarbituric acid reactive substances (TBARS) with the risk of endometriosis in women of childbearing age. Methods: This case-control study enrolled 124 women between January 2018 and July 2019: 35 women with endometriosis (cases) and 89 women without endometriosis undergoing abdominal surgery for other reasons (controls). Endometriosis was diagnosed (cases) or ruled out (controls) by laparoscopic inspection of the pelvis and the biopsy of suspected lesions (histological diagnosis). Fasting urine samples were collected before surgery to determine concentrations of BPA, BPS, BPF, and TBARS. Associations of bisphenol and TBARS concentrations with endometriosis risk were explored with multivariate logistic and linear regression analyses. Results: After adjustment for urinary creatinine, age, BMI, parity, and residence, endometriosis risk was increased with each 1 log unit of BPA [OR 1.5; 95%CI 1.0-2.3] and Σbisphenols [OR 1.5; 95%CI 0.9-2.3] but was not associated with the presence of BPS and BPF. Classification of the women by tertiles of exposure revealed statistically significant associations between endometriosis risk and the second tertile of exposure to BPA [OR 3.7; 95%CI 1.3-10.3] and Σbisphenols [OR 5.4; 95%CI 1.9-15.6]. In addition, TBARS concentrations showed a close-to-significant relationship with increased endometriosis risk [OR 1.6; 95%CI 1.0-2.8], and classification by TBARS concentration tertile revealed that the association between endometriosis risk and concentrations of BPA [OR 2.0; 95%CI 1.0-4.1] and Σbisphenols [OR 2.2; 95%CI 1.0-4.6] was only statistically significant for women in the highest TBARS tertile (>4.23 μM). Conclusion: Exposure to bisphenols may increase the risk of endometriosis, and oxidative stress may play a crucial role in this association. Further studies are warranted to verify these findings.


Sexual selection and mating advantages in the giant sperm species, Drosophila bifurca.
Mate choice may be exercised by either sex; however, females are generally choosier than males because they invest more in their gametes. Female choice is often based on direct benefits, such as better reproductive output, whereas male choice appears to be based on differences in female fecundity. However, when gamete production is limited, sexual selection theories predict that mate choice will be decisive for reproductive success in both sexes. Here, we investigate how mating advantage is achieved in Drosophila bifurca, a giant sperm species in which both sexes produce only a few gametes. Our initial expectations were as follows: (1) females would discriminate against sperm-depleted males to avoid fertility cost; and (2) males would discriminate against inseminated females to reduce sperm competition and increase the assurance of paternity of individual gametes. Differences in courtship behaviors were analyzed with regard to the sexual maturity, which is reached after 22 days in males at 21 degrees C, and the reproductive history of both sexes (inseminated versus virgin for females, and sperm-depleted versus sperm-loaded for males). Our results show that: (1) sexual immaturity precludes mating in both sexes; (2) virgin females do not discriminate between sperm-loaded and sperm-depleted males, and (3) males mate preferentially with virgin females, because inseminated females fend off the male, which tended to bring male courtship to an end. Female remating was limited, but increased significantly when the first male was sperm-depleted. Contrary to our initial expectations, these findings suggest that male sperm depletion does not affect female mating preference, whereas the success of male courtship is driven by female behavior. The possibility that female remating was only promoted in response to low sperm transfer is discussed in relation to the gametic system of this species.


Pairwise comparisons of mitochondrial DNA sequences in subdivided populations and implications for early human evolution.
We consider the effect on the distribution of pairwise differences between mitochondrial DNA sequences of the incorporation into the underlying population genetics model of two particular effects that seem realistic for human populations. The first is that the population size was roughly constant before growing to its current level. The second is that the population is geographically subdivided rather than panmictic. In each case these features tend to encourage multimodal distributions of pairwise differences, in contrast to existing, unimodal datasets. We argue that population genetics models currently used to analyze such data may thus fail to reflect important features of human mitochondrial DNA evolution. These may include selection on the mitochondrial genome, more realistic mutation mechanisms, or special population or migration dynamics. Particularly in view of the variability inherent in the single available human mitochondrial genealogy, it is argued that until these effects are better understood, inferences from such data should be rather cautious.


Methicillin-resistant Staphylococcus aureus bacteraemia and endocarditis treated with ceftaroline salvage therapy.
One of the newest methicillin-resistant Staphylococcus aureus (MRSA) antibiotics to receive FDA approval is ceftaroline fosamil, a member of a new subclass of cephalosporins with unique activity against MRSA. However, ceftaroline is currently only FDA approved for complicated skin/soft tissue infections and community-acquired pneumonia; there are currently no clinical data regarding its use in MRSA bacteraemia and endocarditis. We report a series of six patients in which ceftaroline was utilized as salvage monotherapy in persistent MRSA bacteraemia or endocarditis. Using pharmacy records, 11 ceftaroline-treated patients were identified between January 2011 and November 2011 at University Health System and the South Texas Veterans Health Care System in San Antonio, TX, USA. All cases were reviewed and six patients received ceftaroline therapy for MRSA bacteraemia or endocarditis due to persistent or recurrent bacteraemia while on standard antibiotics (vancomycin or daptomycin). All six patients experienced rapid clearance of their bacteraemia after starting ceftaroline. In the case of endocarditis for which the patient subsequently developed heart failure and required valve replacement, there was no evidence of growth from cultures taken from the excised valve, suggesting sterilization within 13 days of starting ceftaroline. Ceftaroline exhibits potent anti-MRSA activity in both in vitro and animal studies, including rabbit endocarditis models; however, the lack of clinical data has limited its use in bacteraemia and endovascular infections in humans. We hope that this series serves as an initial stepping stone for further evaluation of this compound for more invasive infections due to MRSA.


Familial dilated cardiomyopathy in the United Kingdom.
To determine the frequency and mode of inheritance of familial dilated cardiomyopathy in the United Kingdom. Two recent prospective studies have suggested that familial forms of dilated cardiomyopathy are common but have been limited by selective screening methods, inadequate diagnostic criteria, and low rates of ascertainment. Prospective screening study of 236 relatives from 40 families of patients with dilated cardiomyopathy. Screening consisted of clinical examination, 12 lead electrocardiogram, and two-dimensional Doppler echocardiography. Relatives with systemic hypertension and other cardiac diseases were excluded from the study. All echocardiograms were performed by an experienced echocardiographer who was blinded to clinical information. Relatives were classified as having dilated cardiomyopathy, left ventricular enlargement (method of Henry), depressed fractional shortening, or as being normal. Relatives with abnormal investigations underwent further evaluation as appropriate. Twenty five cases of dilated cardiomyopathy were identified and came from 10 (25%) of the 40 families screened. Pedigree analysis was most consistent with autosomal dominant inheritance and variable penetrance (65-95%). Of the remaining apparently healthy relatives, 37 (18%) were found to have left ventricular enlargement and nine (4%) depressed fractional shortening; these values were significantly higher than those observed in 239 healthy controls (24 (10%), P = 0.02 and one (0.4%), P = 0.01, respectively). Patients with dilated cardiomyopathy commonly have an affected family member and a high proportion of apparently healthy relatives with minor echocardiographic abnormalities. Segregation analysis suggests that familial dilated cardiomyopathy is the result of the transmission of a rare autosomal dominant gene. Further studies are currently underway to characterise the molecular basis of familial dilated cardiomyopathy and identify early disease within these families.


An 86-year-old woman with cardiac cachexia contemplating the end of her life: review of hospice care.
Mrs H is an 86-year-old woman with progressive congestive heart failure and multiple chronic conditions who is experiencing worsening function and quality of life despite maximum medical therapies. She seeks advice regarding control over the circumstances of the end of her life, be it by suicide or under hospice care. Typical of US populations older than 65 years with multiple chronic health conditions and functional decline who are facing the end of life, Mrs H's concerns are particularly about quality of life, not being a burden on loved ones, and maintaining control. As she demonstrates, psychological, existential, and social factors, particularly fear of being a burden, are more common reasons for desiring hastened death than those related directly to physical symptoms. Hospice, which provides a multidisciplinary approach to care at the end of life, can assist Mrs H and her family in making decisions that are consistent with her goals.


Factors modulating preferences of female house mice for males differing in t-complex genotype: role of t-complex genotype, genetic background, and estrous condition of females.
The present study investigates factors mediating odor and social preferences in female house mice (Muc domesticus) based on the t-complex genotype of males. Previous studies in this laboratory showed that females prefer the odors of wild-type (+/+) males over those that carry lethal genes (+/t). The purpose of this study was to determine the genetic, hormonal, and environmental factors that regulate these preferences. The variables examined include t-complex genotype, genetic background, estrous condition of test females (+/+ or +/t), and genotype of the test female's parents. Results indicate that female preferences for +/+ males are dependent upon the t-complex genotype and estrous condition of test females. Only +/t females in estrus showed preferences for +/+ males. Estrous cycle condition effects were seen in both wild (+/+w5) and inbred (129 +/tw5)females. Homozygous females (+/+) and diestrous females of both genotypes did not demonstrate preferences for either +/+ or +/t males.


Effects of berberine and barberry on anthropometric measures: A systematic review and meta-analysis of randomized controlled trials.
Despite controversies, no study has systematically summarized findings from earlier studies on the effect of berberine and barberry on anthropometric measures. Therefore, the current systematic review and meta-analysis was conducted on the effect of berberine and barberry on body mass index (BMI), body weight (BW), waist circumference (WC) and waist-hip ratio (WHR) in adults. Relevant studies, published up to August 2019, were searched through PubMed/Medline, Scopus, ISI Web of Science, Embase and Google Scholar. All randomized clinical trials investigating the effect of berberine and barberry on the anthropometric measures including BMI, BW, WC or/and WHR were included. Out of 252 citations, 12 trials that enrolled 849 subjects were included. Berberine and barberry resulted in no significant change in BMI (Weighted mean differences (WMD): -0.16 kg/m2; 95 % CI: -0.43 to 0.11, P = 0.247), BW (WMD: -0.11 kg; 95 % CI: -0.13 to 0.91, P = 0.830), and berberine resulted in not significant in WC (WMD: -0.58 cm; 95 % CI: -1.89 to 0.72, P = 0.379) and significant reduction in WHR (WMD: -0.03; 95 % CI: -0.04 to -0.01, P < 0.0001). We found a significant reduction in WHR following berberine consumption in adults. Further clinical trials with high quality according to challenges mentioned seem to be helpful to use berberine and barberry as a supplement for certain health conditions, efficiently.


Investigation of optimal sample preparation conditions with potassium triiodide and optimal imaging settings for microfocus computed tomography of excised cat hearts.
To determine optimal sample preparation conditions with potassium triiodide (I2KI) and optimal imaging settings for microfocus CT (micro-CT) of excised cat hearts. 7 excised hearts (weight range, 10 to 17.6 g) obtained from healthy adult cats after euthanasia by IV injection of pentobarbital sodium. Following excision, the hearts were preserved in 10% formaldehyde solution. Six hearts were immersed in 1.25% I2KI solution (n = 3) or 2.5% I2KI solution (3) for a 12-day period. Micro-CT images were acquired at time 0 (prior to iodination) then approximately every 24 and 48 hours thereafter to determine optimal sample preparation conditions (ie, immersion time and concentration of I2KI solution). Identified optimal conditions were then used to prepare the seventh heart for imaging; changes in voltage, current, exposure time, and gain on image quality were evaluated to determine optimal settings (ie, maximal signal-to-noise and contrast-to-noise ratios). Images were obtained at a voxel resolution of 30 μm. A detailed morphological assessment of the main cardiac structures of the seventh heart was then performed. Immersion in 2.5% I2KI solution for 48 hours was optimal for sample preparation. The optimal imaging conditions included a tube voltage of 100 kV, current of 150 μA, and exposure time of 354 milliseconds; scan duration was 12 minutes. Results provided an optimal micro-CT imaging protocol for excised cat hearts prepared with I2KI solution that could serve as a basis for future studies of micro-CT for high resolution 3-D imaging of cat hearts.


3D printed microfluidic devices: enablers and barriers.
3D printing has the potential to significantly change the field of microfluidics. The ability to fabricate a complete microfluidic device in a single step from a computer model has obvious attractions, but it is the ability to create truly three dimensional structures that will provide new microfluidic capability that is challenging, if not impossible to make with existing approaches. This critical review covers the current state of 3D printing for microfluidics, focusing on the four most frequently used printing approaches: inkjet (i3DP), stereolithography (SLA), two photon polymerisation (2PP) and extrusion printing (focusing on fused deposition modeling). It discusses current achievements and limitations, and opportunities for advancement to reach 3D printing's full potential.


Mechanotransduction in bone does not require a functional cyclooxygenase-2 (COX-2) gene.
COX-2 is a key enzyme involved in the response of bone to loading. However, using mice with a null mutation of the COX-2 gene, we found that a functional COX-2 gene is not required for mechanotransduction. This paradoxical finding may have resulted, in part, from mechanically induced COX-1 activity. Cyclooxygenase-2 (COX-2) is an important mediator in the response of bone to mechanical loading, with pharmacological inhibition of COX-2 effectively eliminating or reducing mechanically induced bone formation. In this study, we further investigated the role of COX-2 in skeletal mechanotransduction using a genetic approach. The aim was to compare the skeletal responsiveness of COX-2 homozygous mutant (COX-2(-/-)) and wildtype control (COX-2(+/+)) mice to investigate whether a functional COX-2 gene is necessary for mechanotransduction. Adult female COX-2(+/+) and COX-2(-/-) mice on a C57BL/6x129/ola background were studied using the ulna axial loading model. The response to 2 days of loading for 120 cycles/day at 2 Hz was measured histomorphometrically. Phenotypic characterization of the femurs in these mice was also performed. In a separate group of animals, the expression of the remaining COX isozyme, COX-1, was assessed using real-time RT-PCR 4 h after one bout of 120 loading cycles. Null mutation of the COX-2 gene resulted in a consistent femoral phenotype of reduced bone mass, altered architecture, and inferior mechanical properties. Many of these differences were nullified after adjustment for body weight. Nevertheless, body weight-corrected values showed a consistent trend of reduced mechanical properties in COX-2(-/-) mice. Genotype did not influence the response to mechanical loading, with no histomorphometric differences being found between COX-2(+/+) and COX-2(-/-) mice. Real-time RT-PCR showed COX-2(-/-) mice to express significantly greater COX-1 expression in loaded ulnas than in loaded ulnas in COX-2(+/+) mice. There were no differences in COX-1 expression in nonloaded ulnas. A functional COX-2 gene was not found to be required for skeletal mechanotransduction. This is in contrast to previous pharmacological studies showing that COX-2 is critical to the response of bone to loading. Investigating a potential reason for the absence of a genotype difference in this study, we found that mice with a null mutation in the COX-2 gene possess inductive skeletal COX-1 expression.


Is the elusive Gymnothorax polyuranodon really a freshwater moray?
Analysis of 36 records of the rarely encountered moray Gymnothorax polyuranodon indicate that juveniles and adults inhabit fresh and mildly brackish habitats (salinity < 5) in streams of the Australian Wet Tropics Eighty-one per cent of these records were from freshwater streams and collectively demonstrate that this species inhabits fresh water throughout all seasons. A survey of fish researchers, each with at least 100 h of field experience in Australia's Wet Tropics, revealed that 33% of researchers working in fresh waters (nine of 27 researchers) had encountered the species and 15% of researchers with substantial experience working in estuaries (two of 13 researchers) had encountered the species. The species was not sampled or observed in the nearshore marine environment. The only record of an elver of this species was, however, found in an estuary at a salinity of 33·4. This preliminary evidence suggests adult G. polyuranodon occupy freshwater habitats, but further research is required to understand the complete life cycle, including movements, habitat use and reproductive ecology of the species.


Moderation of a parent-based intervention on transitions in drinking: examining the role of normative perceptions and attitudes among high- and low-risk first-year college students.
Parent-based interventions (PBIs) are an effective strategy to reduce problematic drinking among first-year college students. The current study examined the extent to which student-based characteristics, derived from the Theory of Planned Behavior, moderated 3 PBI conditions: (i) prior to college matriculation (PCM); (ii) PCM with a booster during the fall semester; and (iii) after college matriculation. The moderator variables included injunctive and descriptive peer norms about alcohol use and attitudes toward alcohol use. Using data from a randomized control trial delivered to 1,900 incoming college students, we examined differential treatment effects within 4 types of baseline student drinkers: (i) nondrinkers; (ii) weekend light drinkers (WLD); (iii) weekend heavy episodic drinkers; and (iv) heavy drinkers. The outcome variable was based on the transitions in drinking that occurred between the summer prior to college enrollment and the end of the first fall semester and distinguished between students who transitioned to 1 of the 2 risky drinking classes. The results indicated that injunctive norms (but not descriptive norms or attitudes) moderated the differential effects of the PBI with strongest effects for students whose parents received the booster. Differential effects also depended on baseline drinking class and were most pronounced among WLDs who were deemed "high-risk" in terms of injunctive peer norms. Parental influence can remain strong for young adults who are transitioning to college environments, even among students with relatively high peer influence to drink alcohol. Thus, the PBI represents an effective tool to prevent escalation of alcohol use during the first year of college, when risk is highest and patterns of alcohol use are established.


The role of donor age and gender in the success of human muscle precursor cell transplantation.
Autologous cell transplantation for the treatment of muscle damage is envisioned to involve the application of muscle precursor cells (MPCs) isolated from adult skeletal muscle. At the onset of trauma, these cells are recruited to proliferate and rebuild injured muscle fibres. However, a variety of donor-specific cues may directly influence the yield and quality of cells isolated from a muscle biopsy. In this study, we isolated human MPCs and assessed the role of donor gender and age on the ability of these MPCs to form functional bioengineered muscle. We analysed the cell yield, growth and molecular expression in vitro, and the muscle tissue formation and contractility of the bioengineered muscle, from cells isolated from men and women in three different age groups: young (20-39 years), adult (40-59 years) and elderly (60-80 years). Our results suggest that human MPCs can be successfully isolated and grown from patients of all ages and both genders. However, young female donors provide fast-growing cells in vitro with an optimum contractile output in vivo and are therefore an ideal cell source for muscle reconstruction. Taken together, these findings describe the donor-related limitations of MPC transplantation and provide insights for a straightforward and unbiased clinical application of these cells for muscle reconstruction. Copyright © 2014 John Wiley & Sons, Ltd.


Chromatin-bound cGAS is an inhibitor of DNA repair and hence accelerates genome destabilization and cell death.
DNA repair via homologous recombination (HR) is indispensable for genome integrity and cell survival but if unrestrained can result in undesired chromosomal rearrangements. The regulatory mechanisms of HR are not fully understood. Cyclic GMP-AMP synthase (cGAS) is best known as a cytosolic innate immune sensor critical for the outcome of infections, inflammatory diseases, and cancer. Here, we report that cGAS is primarily a chromatin-bound protein that inhibits DNA repair by HR, thereby accelerating genome destabilization, micronucleus generation, and cell death under conditions of genomic stress. This function is independent of the canonical STING-dependent innate immune activation and is physiologically relevant for irradiation-induced depletion of bone marrow cells in mice. Mechanistically, we demonstrate that inhibition of HR repair by cGAS is linked to its ability to self-oligomerize, causing compaction of bound template dsDNA into a higher-ordered state less amenable to strand invasion by RAD51-coated ssDNA filaments. This previously unknown role of cGAS has implications for understanding its involvement in genome instability-associated disorders including cancer.


Estimated herd prevalence and sequence types of Coxiella burnetii in bulk tank milk samples from commercial dairies in Indiana.
Coxiella burnetii is the etiologic agent of Q fever, a zoonotic disease causing influenza-like illness, pregnancy loss, cardiovascular disease and chronic fatigue syndrome in people. C. burnetii is considered to be enzootic in ruminants, but clinical signs of infection do not always manifest. National studies have documented the presence of C. burnetii in dairy herds in Indiana. This represents an opportunity to better characterize the distribution and prevalence of C. burnetii infection at the state scale, allowing evaluation of the need for surveillance and response planning to occur at this level. A cross-sectional study was conducted to estimate the herd prevalence of C. burnetii in commercial cattle dairies in Indiana and characterize the strains of C. burnetii within these dairies. Bulk tank milk samples were collected between June and August of 2011 by the Indiana State Board of Animal Health (ISBOAH). A total of 316 of these samples were tested for the IS1111 transposon of C. burnetii using quantitative real time polymerase chain reaction (PCR). Single nucleotide polymorphism (SNP) genotyping was used to identify the multispacer sequence genotypes (ST) present in samples where the IS1111 transposon was identified. The geographic distribution of dairies testing positive for C. burnetii DNA and the identified STs were also evaluated. The estimated overall herd prevalence for C. burnetii DNA was 61.1 % (95 % CI 55.6-66.3 %). The highest estimated regional prevalence was 70.2 % in the Central region of Indiana. An ST was identifiable in 74 of the positive 178 samples (41.6 %) and none of the 10 negative samples tested. Of these samples, 71 (95.9 %) were identified as ST20, 2 (2.7 %) as ST8 and a combination of ST20 and ST8 was identified in a single sample. C. burnetii is present in dairy herds throughout Indiana. Indiana follows national trends with ST20 most commonly identified. The presence of multiple STs in a single bulk tank sample indicates that multiple strains of C. burnetii can circulate within a herd. This supports potential transmission of C. burnetii between goats and cattle, presenting the potential for a switch in the dominant genotype found in a given species.


Venous smooth muscle in hypertension. Enhanced contractility of portal veins from spontaneously hypertensive rats.
In isolated bath studies smooth muscle from the rat portal vein was evaluated for its reactivity and contractility, and the whole vessel wall was evaluated for its extensibility. Smooth muscle from the spontaneously hypertensive rat (SHR) had the following characteristics when compared with that from normotensive controls: (1) Spontaneous phasic contractions were more frequent and developed more tension; (2) threshold concentrations for responses to prostaglandins A-2 and B-2 were lower, but those for responses to epinephrine, norepinephrine, KCl, BaCl-2, and SrCl-2 were similar; (3) high concentrations of calcium had a less depressant action on the responses to the prostaglandins but not on the responses to the other agonists; (4) maximal contractile tensions to all agonists were greater; and (5) passive extensibility was less. These differences, because they are in the venous system, cannot be secondary to the increase in wall stress of arterial hypertension. The decreased passive extensibility in this vein in SHR creates a stiffer framework on which the active contractile process is able to develop greater tension. If this increase in active tension is generalized to all veins, it could be responsible for a decrease in venous capacity which increases the rate of venous return and, hence, increases cardiac output.


[A successful case of lower gingival cancer with pulmonary metastases by adjuvant chemotherapy including paclitaxel, cisplatin and 5-fluorouracil following a surgical procedure].
We report a successful case with pulmonary metastases from lower gingival cancer by a surgical procedure and four cycles of adjuvant chemotherapy including paclitaxel (PTX), cisplatin (CDDP) and 5-fluorouracil (5-FU). A 47-year-old woman underwent chemotherapy with CDDP and 5-FU after an operation for lower gingival squamous cell carcinoma and its neck lymph node metastases. At 4 months from the initial treatment, pulmonary metastatic lesion was resected by video-assisted thoracoscopic surgery (VATS). Fourteen months later, pulmonary metastatic lesion was found and dissected again using VATS. Furthermore, the patient was treated by adjuvant chemotherapy with PTX 135 mg/m(2) over 3 hours on day 1, CDDP 75 mg/m(2)on day 2 and 5-FU 350 mg/m(2)/day by continuous intravenous infusion on day 2 through 5. After that, there is no evidence of pulmonary recurrence for more than six years.


Palliation of dysphagia.
Palliation of dysphagia is the cornerstone of palliative treatment in patients with incurable oesophageal cancer. Available palliative options for dysphagia are oesophageal stent placement and radiotherapy. In general, oesophageal stent placement is the preferred therapeutic option in patients with a relatively poor prognosis because of its rapid relief of dysphagia. Regardless of ongoing technical developments, recurrence of dysphagia and stent-related complications are still occurring. For patients with a relatively good prognosis, intra-luminal brachytherapy is advised because of its sustained palliation of dysphagia. Due to limited availability of intra-luminal brachytherapy in clinical practice, fractionated external beam radiation therapy is commonly applied as an alternative. Selection of the optimal palliative approach for patients remains however challenging as conclusive high-quality evidence is limited. Moreover, with the introduction of new palliative treatment options (e.g. palliative chemotherapeutic and radiotherapeutic options) and the concurrent change of patient characteristics, supporting evidence from large randomised studies is warranted.


Activation of Fetal γ-globin Gene Expression via Direct Protein Delivery of Synthetic Zinc-finger DNA-Binding Domains.
Reactivation of γ-globin expression has been shown to ameliorate disease phenotypes associated with mutations in the adult β-globin gene, including sickle cell disease. Specific mutations in the promoter of the γ-globin genes are known to prevent repression of the genes in the adult and thus lead to hereditary persistence of fetal hemoglobin. One such hereditary persistence of fetal hemoglobin is associated with a sequence located 567 bp upstream of the Gγ-globin gene which assembles a GATA-containing repressor complex. We generated two synthetic zinc-finger DNA-binding domains (ZF-DBDs) targeting this sequence. The -567Gγ ZF-DBDs associated with high affinity and specificity with the target site in the γ-globin gene promoter. We delivered the -567Gγ ZF-DBDs directly to primary erythroid cells. Exposure of these cells to the recombinant -567Gγ ZF-DBDs led to increased expression of the γ-globin gene. Direct protein delivery of ZF-DBDs that compete with transcription regulatory proteins will have broad implications for modulating gene expression in analytical or therapeutic settings.


Permeability and electrokinetic characterization of poly(ethylene terephthalate) capillary pore membranes with grafted temperature-responsive polymers.
Poly(ethylene terephthalate) (PET) track-etched membranes with average pore diameters of 692 and 1629 nm were functionalized using the monomer N-isopropylacrylamide (NIPAAm) and a photoinitiated "grafting-from" approach in which a surface-selective reaction has been most efficiently achieved by combinations of the unmodified PET surface with benzophenone and, alternatively, of an aminated PET surface with benzophenone carboxylic acid. Consistent estimations of the pore diameters of the base PET membranes and of the effective grafted polyNIPAAm layer thicknesses on the PET pore walls were possible only on the basis of the permeabilities measured with aqueous solutions of higher ionic strength (e.g., 0.1 M NaCl). However, the permeabilities measured with ultrapure water indicated that the "electroviscous effect" was significant for both base membranes. The influences of membrane pore diameter, surface charge, and solution ionic strength could be interpreted in the framework of the space-charge model. Functionalized membranes with collapsed grafted polymer hydrogel layer thicknesses of a few nanometers exhibited almost zero values of the zeta potential estimated from the trans-membrane streaming potential measurements. This was caused by a "hydrodynamic screening" of surface charge by the neutral hydrogel. Very pronounced changes in permeability as a function of temperature were measured for PET membranes with grafted polyNIPAAm layers, and the effective layer thickness in the swollen state--here up to approximately 300 nm--correlated well with the degree of functionalization. The subtle additional effects of solution ionic strength on the hydrodynamic layer thickness at 25 degrees C were different from the effects for the base PET membranes and could be explained by a variation in the degree of swelling, resembling a "salting-out" effect. Overall, it had been demonstrated that the functionalized capillary pore membranes are well suited for a detailed and quantitative evaluation of the relationships between the synthesis, the structure, and the function of grafted stimuli-responsive polymer layers.


Stereotactic radiotherapy in oligometastatic cancer.
Oligometastatic cancer describes a disease state somewhere between localized and metastatic cancer. Proposed definitions of oligometastatic disease have typically used a cut-off of five or fewer sites of disease. Treatment of oligometastatic disease should have the goal of long-term local control, and in selected cases, disease remission. While several retrospective cohorts argue for surgical excision of limited metastases (metastasectomy) as the preferred treatment option for several clinical indications, limited randomized data exists for treating oligometastases. Alternatively, stereotactic ablative radiotherapy (SABR) is a radiotherapy technique that combines high radiation doses per fraction with precision targeting with the goal of achieving long-term local control of treated sites. Published cohort studies of SABR have demonstrated excellent local control rates of 70-90% in oligometastatic disease, with long-term survival in some series approaching 20-40%. A recent randomized phase 2 clinical trial by Gomez et al. demonstrated significantly improved progression free survival with aggressive consolidative therapy (surgery, radiotherapy ± chemotherapy or SABR) in oli-gometastatic non-small cell lung cancer (NSCLC). As additional randomized controlled trials are ongoing to determine the efficacy of SABR in oligometastatic disease, SABR is increasingly being used within routine clinical practice. This review article aims to sum-marize the history and current paradigm of the oligometastatic state, review recently pub-lished literature of SABR in oligometastatic cancer and discuss ongoing trials and future directions in this context.


Macrolide therapy suppresses key features of experimental steroid-sensitive and steroid-insensitive asthma.
Steroid-insensitive endotypes of asthma are an important clinical problem and effective therapies are required. They are associated with bacterial infection and non-eosinophilic inflammatory responses in the asthmatic lung. Macrolide therapy is effective in steroid-insensitive endotypes, such as non-eosinophilic asthma. However, whether the effects of macrolides are due to antimicrobial or anti-inflammatory mechanisms is not known. To determine and assess the efficacy of macrolide (ie, clarithromycin) and non-macrolide (ie, amoxicillin) antibiotic treatments in experimental models of infection-induced, severe, steroid-insensitive neutrophilic allergic airways disease (SSIAAD), compared with steroid-sensitive AAD and to delineate the antimicrobial and anti-inflammatory effects of macrolide therapy. We developed and used novel mouse models of Chlamydia and Haemophilus lung infection-induced SSIAAD. We used these models to investigate the effects of clarithromycin and amoxicillin treatment on immune responses and airways hyper-responsiveness (AHR) in Ova-induced, T helper lymphocyte (Th) 2 -associated steroid-sensitive AAD and infection-induced Th1/Th17-associated SSIAAD compared with dexamethasone treatment. Clarithromycin and amoxicillin had similar antimicrobial effects on infection. Amoxicillin did attenuate some features, but did not broadly suppress either form of AAD. It did restore steroid sensitivity in SSIAAD by reducing infection. In contrast, clarithromycin alone widely suppressed inflammation and AHR in both steroid-sensitive AAD and SSIAAD. This occurred through reductions in Th2 responses that drive steroid-sensitive eosinophilic AAD and tumour necrosis factor α and interleukin 17 responses that induce SSIAAD. Macrolides have broad anti-inflammatory effects in AAD that are likely independent of their antimicrobial effects. The specific responses that are suppressed are dependent upon the responses that dominate during AAD.


Executive dysfunction predicts social cognition impairment in amyotrophic lateral sclerosis.
Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder of the motor system with recognised extra-motor and cognitive involvement. This cross-sectional study examined ALS patients' performance on measures requiring social inference, and determined the relationship between such changes and variations in mood, behaviour, personality, empathy and executive function. Fifty-five ALS patients and 49 healthy controls were compared on tasks measuring social cognition and executive function. ALS patients also completed measures examining mood, behaviour and personality. Regression analyses explored the contribution of executive function, mood, behaviour and personality to social cognition scores within the ALS sample. A between-group MANOVA revealed that, the ALS group was impaired relative to controls on two composite scores for social cognition and executive function. Patients also performed worse on individual tests of executive function measuring cognitive flexibility, response inhibition and concept formation, and on individual aspects of social cognition assessing the attribution of emotional and mental states. Regression analyses indicated that ALS-related executive dysfunction was the main predictor of social cognition performance, above and beyond demographic variables, behaviour, mood and personality. On at least some aspects of social cognition, impaired performance in ALS appears to be secondary to executive dysfunction. The profile of cognitive impairment in ALS supports a cognitive continuum between ALS and frontotemporal dementia.


Group cognitive behavioural treatment for insomnia in primary care: a randomized controlled trial.
Insomnia disorder is common and often co-morbid with mental health conditions. Cognitive behavioural therapy (CBT) for insomnia is effective, but is rarely implemented as a discrete treatment. The aim of this study was to evaluate the effectiveness of brief CBT groups for insomnia compared to treatment as usual (TAU) for insomnia delivered by mental health practitioners in a primary-care mental health service. A total of 239 participants were randomized to either a five-session CBT group or to TAU. Assessments of sleep and of symptoms of depression and anxiety were carried out at baseline, post-treatment and at 20 weeks. Primary outcome was sleep efficiency post-treatment. Group CBT participants had better sleep outcomes post-treatment than those receiving TAU [sleep efficiency standardized mean difference 0.63, 95% confidence interval (CI) 0.34-0.92]. The effect at 20 weeks was smaller with a wide confidence interval (0.27, 95% CI -0.03 to 0.56). There were no important differences between groups at either follow-up period in symptoms of anxiety or depression. Dedicated CBT group treatment for insomnia improves sleep more than treating sleep as an adjunct to other mental health treatment.


Contralateral Hippocampal Stimulation for Failed Unilateral Anterior Temporal Lobectomy in Patients with Bilateral Temporal Lobe Epilepsy.
To prospectively study the surgical outcomes of unilateral anterior temporal lobectomy (ATL) in patients with intractable bilateral temporal lobe epilepsy (TLE) as well as two-staged contralateral hippocampal stimulation in patients after failed unilateral ATL. Eighteen carefully selected patients with bilateral TLE underwent unilateral ATL. Five cases with failed ATL underwent two-staged contralateral hippocampal stimulation. Seizure control and changes in intelligence quotient (IQ), memory quotient, and quality of life (QOL) were analyzed 2-5 years after treatment. In the patients with unilateral ATL, the percentages seizure free were 55.6% (10/18), 50.0% (9/18), and 44.4% (4/9) at the 1-, 2-, and 5-year follow-up visits, respectively. There were significant difference in seizure control between the patients with unilateral ATL and the 12 cases in the medication group. Significant differences were also found in changes in the patients' QOL and full-scale IQ at the 2-year follow-up between the surgical and medication groups. Five patients who underwent contralateral hippocampal stimulation after failed unilateral ATL experienced 80-100% seizure reductions, and 80% were seizure free 1 year after hippocampal stimulation. Unilateral ATL provides good seizure control and does not cause serious memory or IQ injury in carefully selected patients with true bilateral TLE. Contralateral hippocampal stimulation is a useful approach for patients who experience unilateral ATL failure.


MObjects--a novel method for the visualization and interactive exploration of defects in industrial XCT data.
This paper describes an advanced visualization method for the analysis of defects in industrial 3D X-Ray Computed Tomography (XCT) data. We present a novel way to explore a high number of individual objects in a dataset, e.g., pores, inclusions, particles, fibers, and cracks demonstrated on the special application area of pore extraction in carbon fiber reinforced polymers (CFRP). After calculating the individual object properties volume, dimensions and shape factors, all objects are clustered into a mean object (MObject). The resulting MObject parameter space can be explored interactively. To do so, we introduce the visualization of mean object sets (MObject Sets) in a radial and a parallel arrangement. Each MObject may be split up into sub-classes by selecting a specific property, e.g., volume or shape factor, and the desired number of classes. Applying this interactive selection iteratively leads to the intended classifications and visualizations of MObjects along the selected analysis path. Hereby the given different scaling factors of the MObjects down the analysis path are visualized through a visual linking approach. Furthermore the representative MObjects are exported as volumetric datasets to serve as input for successive calculations and simulations. In the field of porosity determination in CFRP non-destructive testing practitioners use representative MObjects to improve ultrasonic calibration curves. Representative pores also serve as input for heat conduction simulations in active thermography. For a fast overview of the pore properties in a dataset we propose a local MObjects visualization in combination with a color-coded homogeneity visualization of cells. The advantages of our novel approach are demonstrated using real world CFRP specimens. The results were evaluated through a questionnaire in order to determine the practicality of the MObjects visualization as a supportive tool for domain specialists.


Reparative radiological changes of hip joint after TNF inhibitors in ankylosing spondylitis.
Hip involvement in ankylosing spondylitis (AS) is a common extraspinal arthritic manifestation, which is associated to a worse functional outcome. Little data are available on the effectiveness of conservative treatment strategies. The TNF inhibitors have been proven effective on AS activity parameters. Their structural effect on hip disease however, little is studied. We describe four new cases of reparative changes of a damaged hip joint after treatment with TNF inhibitors. The average of age was 32.5 (27- 36) years. There were 3 men and 1 woman. Hip involvement was bilateral in all cases. Etanercept was prescribed in 3 cases and infliximab in 1 case. At baseline, all patients had a painful and limited hip with high disease activity and an important functional impairment. After an average of 5.5 years of treatment with TNF inhibitors, the BASRI hip evaluated in antero-posterior x-rays of the pelvis remained unchanged at 2.4. The average of mean hip joint space was 2.9mm (2.3-3.6). A widening in hip joint space was observed in all cases with less subchondral cysts. TNF inhibitors seem to be effective on hip joint disease in patients with AS.


Adrenoleukodystrophy in female heterozygotes: underrecognized and undertreated.
X-linked adrenoleukodystrophy (X-ALD) is a neurodegenerative disease resulting from mutations in the gene ABCD1 and alterations in peroxisomal beta-oxidation of long chain fatty acids. As it has been frequently discussed, it manifests a wide range of phenotypes in male, with progressive myelopathy being the most common. Even though the gene is localized to the X-chromosome and a region subject to X-inactivation, female carriers still are affected significantly by this condition. It has been stated that between 20 and 50% of women who are carriers may manifest some symptoms and recent evidence has suggested the differences in disease manifestations and relative rates of progression between men and women. However there have been only limited studies specifically addressing this and to date there has been no comprehensive review discussing the different phenotypes in female carriers, as well as the differences in disease onset, progression, disability, nervous system pathology and neuroimaging patterns compared to affected males. This is of key importance as similarities and differences between genders will assist in determining how best to target therapies in all affected individuals as opportunities for treatment present themselves. As will be further addressed in this review, we need to improve our understanding of the associations of emergent neuroimaging techniques to physical disability in this population. We reviewed the clinical presentations in the carrier population, the distinct disability profile and neuroimaging findings in order to put together pieces of this neglected segment in X-ALD and give direction to further studies.


Comprehensive Phylogenetic Analysis of Bovine Non-aureus Staphylococci Species Based on Whole-Genome Sequencing.
Non-aureus staphylococci (NAS), a heterogeneous group of a large number of species and subspecies, are the most frequently isolated pathogens from intramammary infections in dairy cattle. Phylogenetic relationships among bovine NAS species are controversial and have mostly been determined based on single-gene trees. Herein, we analyzed phylogeny of bovine NAS species using whole-genome sequencing (WGS) of 441 distinct isolates. In addition, evolutionary relationships among bovine NAS were estimated from multilocus data of 16S rRNA, hsp60, rpoB, sodA, and tuf genes and sequences from these and numerous other single genes/proteins. All phylogenies were created with FastTree, Maximum-Likelihood, Maximum-Parsimony, and Neighbor-Joining methods. Regardless of methodology, WGS-trees clearly separated bovine NAS species into five monophyletic coherent clades. Furthermore, there were consistent interspecies relationships within clades in all WGS phylogenetic reconstructions. Except for the Maximum-Parsimony tree, multilocus data analysis similarly produced five clades. There were large variations in determining clades and interspecies relationships in single gene/protein trees, under different methods of tree constructions, highlighting limitations of using single genes for determining bovine NAS phylogeny. However, based on WGS data, we established a robust phylogeny of bovine NAS species, unaffected by method or model of evolutionary reconstructions. Therefore, it is now possible to determine associations between phylogeny and many biological traits, such as virulence, antimicrobial resistance, environmental niche, geographical distribution, and host specificity.


Three VCP Mutations in Patients with Frontotemporal Dementia.
Valosin-containing protein (VCP) is involved in multiple cellular activities. Mutations in VCP lead to heterogeneous clinical presentations including inclusion body myopathy with Paget's disease of the bone, frontotemporal dementia and amyotrophic lateral sclerosis, even in patients carrying the same mutation. We screened a cohort of 48 patients with familial frontotemporal dementia (FTD) negative for MAPT, GRN, and C9orf72 mutations for other known FTD genes by using whole exome sequencing. In addition, we carried out targeted sequencing of a cohort of 37 patients with frontotemporal lobar degeneration with Transactive response DNA-binding protein 43 (TDP-43) subtype from the Netherlands Brain bank. Two novel (p.Thr262Ser and p.Arg159Ser) and one reported (p.Met158Val) VCP mutations in three patients with a clinical diagnosis of FTD were identified, and were absence in population-match controls. All three patients presented with behavioral changes, with additional semantic deficits in one. No signs of Paget or muscle disease were observed. Pathological examination of the patient with VCP p.Arg159Ser mutation showed numerous TDP-43 immunoreactive (IR) neuronal intranuclear inclusions (NII) and dystrophic neurites (DN), while a lower number of NII and DN were observed in the patient with the VCP p.Thr262Ser mutation. Pathological findings of both patients were consistent with FTLD-TDP subtype D. Furthermore, only rare VCP-IR NII was observed in both cases. Our study expands the clinical heterogeneity of VCP mutations carriers, and indicates that other additional factors, such as genetic modifiers, may determine the clinical phenotype.


Enhancement of light scattering and photoluminescence in electrospun polymer nanofibers.
Poly(methyl methacrylate) nanofibers with desired fiber diameters that ranged from 336 to 896 nm were electrospun as light scattering and propagation materials. The light scattering behavior of these samples as a function of the fiber diameter and fiber deposition thickness was examined by UV-vis spectrophotometry, which revealed the scattering bands in the absorption spectra. The scattering bands of these nanofibers were linearly proportional to the fiber diameter, which shows good agreement with a scattering model based on the Mie theory. The light scattering and prolonged light path lengths in the nanofiber scaffolds were monitored and quantified by the photoluminescence of a fluorescent dye, Coumarin 6, which was preloaded into the polymer nanofibers. The photoluminescence after proper normalization showed a second-order dependence on the dye loading per unit area, which is significantly different from the spin-coated thin-film samples following a first-order relationship. Nonlinear photoluminescence enhancements indicated prolonged light path lengths and multiple light absorptions within the fiber scaffolds as a result of light scattering. Even with relatively broad scattering band widths, the light scattering and photoluminescence of the electrospun nanofibers exhibited considerable wavelength selectivity, especially as the scattering bands overlapped with the excitation wavelengths of the fluorescence reagent.


Targeting the epidermal growth factor receptor. Trials in head and neck and lung cancer.
The epidermal growth factor receptor (EGFR) promotes the growth of different cell types and has been implicated in tumorigenesis. The EGFR comprises a family of four structurally similar tyrosine kinases with a complex link to downstream signaling molecules that ultimately regulate key cell processes. Anti-EGFR agents have been developed as promising therapeutic anticancer targets, and some have been recently approved for the treatment of non-small-cell lung cancer and colon cancer. The two anti-EGFR therapies with the greatest clinical application are monoclonal antibodies that block the binding of ligands to EGFR and small-molecule tyrosine kinase inhibitors that inhibit the binding of adenosine triphosphate to the internal tyrosine kinase receptor of EGFR. We attempt to give an overview of the EGFR function and biology, focusing on the most important clinical findings and applications of EGFR inhibitors in lung and head and neck cancer.


In-vitro pharmacokinetic/pharmacodynamic model data suggest a potential role of new formulations of posaconazole against Candida krusei but not Candida glabrata infections.
Posaconazole exhibits in-vitro activity against Candida glabrata and Candida krusei. Epidemiological cut-off values set by the European Committee on Antimicrobial Susceptibility Testing (EUCAST) and the Clinical and Laboratory Standards Institute (CLSI) are 1/1 and 0.5/0.5 mg/L, respectively, but clinical breakpoints have not been established to date. This study explored the pharmacodynamics (PD) of posaconazole in a validated one-compartment in-vitro pharmacokinetic (PK)/PD model, and determined the probability of PK/PD target attainment (PTA) for the available formulations. Five C. glabrata and three C. krusei isolates with posaconazole minimum inhibitory concentrations (MICs) of 0.06-2 and 0.03-0.25 mg/L, respectively, were tested in the PK/PD model simulating different time-concentration profiles of posaconazole. The exposure-effect relationship fAUC0-24/MIC was described for EUCAST/CLSI methods, and PTA was calculated in order to determine PK/PD susceptibility breakpoints for oral solution (400 mg q12h), and intravenous (i.v.)/tablet formulations (300 mg q24h). Fungicidal activity (~2log kill) was found against the most susceptible C. glabrata isolate alone, and against all three C. krusei isolates. The corresponding EUCAST/CLSI PK/PD targets (fAUC0-24/MIC) were 102/79 for C. glabrata and 12/8 for C. krusei. Mean PTA was high (>95%) for C. glabrata isolates with EUCAST/CLSI MICs ≤0.03/≤0.03 mg/L for oral solution and ≤0.125/≤0.125 mg/L for i.v. and tablet formulations for the wild-type population. For C. krusei isolates, mean PTA was high (>95%) for EUCAST/CLSI MICs ≤0.25/≤0.5 mg/L for oral solution and ≤1/≤2 mg/L for i.v. and tablet formulations for the wild-type population. The use of posaconazole to treat C. glabrata infections is questionable. Intravenous and tablet formulations may be therapeutic options for the treatment of C. krusei infections, and oral exposure can be optimized with therapeutic drug monitoring (trough levels >0.6-0.9 mg/L).


Treg expression of CIS suppresses allergic airway inflammation through antagonizing an autonomous TH2 program.
Maintenance of regulatory T (Treg) cells is crucial for the regulatory function of Treg cells in immune homeostasis and self-tolerance; however, the detailed underlying mechanisms remain elusive. In the current study, we found that the cytokine suppressor CIS (cytokine induced SH-2 protein) is required for maintenance of Treg cell identity. Mice with Treg-specific Cis-deficiency displayed aggravated experimental allergic asthma, and in adulthood, developed splenomegaly, lymphadenopathy and spontaneous eosinophilic airway inflammation, accompanied by accumulation of effector memory helper T (TH) cells. Cis-deficiency led to the loss of Foxp3 expression and the decrease in suppressive function of Treg cells. Cis-deficient Treg cells expressed TH2 cell signature genes, Gata3, Irf4 and Il4, and excessive interleukin-4-signal transducer and activator of transcription 6 (IL-4-STAT6) signals resulted in repressive chromatin modification in the Foxp3 locus and permissive modification in the Il4 loci. In vitro, blockade of IL-4 restored the expression of Foxp3 and the suppressive function of inducible Treg (iTreg) cells. Thus, we identified a novel feedback loop in stabilization of Treg cells and suppression of TH2-type inflammation in a Treg-intrinsic manner.


2020 AAHA Anesthesia and Monitoring Guidelines for Dogs and Cats.
Risk for complications and even death is inherent to anesthesia. However, the use of guidelines, checklists, and training can decrease the risk of anesthesia-related adverse events. These tools should be used not only during the time the patient is unconscious but also before and after this phase. The framework for safe anesthesia delivered as a continuum of care from home to hospital and back to home is presented in these guidelines. The critical importance of client communication and staff training have been highlighted. The role of perioperative analgesia, anxiolytics, and proper handling of fractious/fearful/aggressive patients as components of anesthetic safety are stressed. Anesthesia equipment selection and care is detailed. The objective of these guidelines is to make the anesthesia period as safe as possible for dogs and cats while providing a practical framework for delivering anesthesia care. To meet this goal, tables, algorithms, figures, and "tip" boxes with critical information are included in the manuscript and an in-depth online resource center is available at aaha.org/anesthesia.


Identification of Cancer-Associated Fibroblasts in Circulating Blood from Patients with Metastatic Breast Cancer.
Metastasis is facilitated by cancer-associated fibroblasts (CAF) in the tumor microenvironment through mechanisms yet to be elucidated. In this study, we used a size-based microfilter technology developed by our group to examine whether circulating CAF identified by FAP and α-SMA co-expression (cCAF) could be distinguished in the peripheral blood of patients with metastatic breast cancer. In a pilot study of patients with breast cancer, we detected the presence of cCAFs in 30/34 (88%) patients with metastatic disease (MET group) and in 3/13 (23%) patients with localized breast cancer (LOC group) with long-term disease-free survival. No cCAFs as defined were detected in healthy donors. Further, both cCAF and circulating tumor cells (CTC) were significantly greater in the MET group compared with the LOC group. Thus, the presence of cCAF was associated with clinical metastasis, suggesting that cCAF may complement CTC as a clinically relevant biomarker in metastatic breast cancer.


Stability of presynaptic vesicle pools and changes in synapse morphology in the amygdala following fear learning in adult rats.
Changes in synaptic strength in the lateral amygdala (LA) that occur with fear learning are believed to mediate memory storage, and both presynaptic and postsynaptic mechanisms have been proposed to contribute. In a previous study we used serial section transmission electron microscopy (ssTEM) to observe differences in dendritic spine morphology in the adult rat LA after fear conditioning, conditioned inhibition (safety conditioning), or naïve control handling (Ostroff et al. [2010] Proc Natl Acad Sci U S A 107:9418-9423). We have now reconstructed axons from the same dataset and compared their morphology and relationship to the postsynaptic spines between the three training groups. Relative to the naïve control and conditioned inhibition groups, the ratio of postsynaptic density (PSD) area to docked vesicles at synapses was greater in the fear-conditioned group, while the size of the synaptic vesicle pools was unchanged. There was significant coherence in synapse size between neighboring boutons on the same axon in the naïve control and conditioned inhibition groups, but not in the fear-conditioned group. Within multiple-synapse boutons, both synapse size and the PSD-to-docked vesicle ratio were variable between individual synapses. Our results confirm that synaptic connectivity increases in the LA with fear conditioning. In addition, we provide evidence that boutons along the same axon and even synapses on the same bouton are independent in their structure and learning-related morphological plasticity.


Pregnancy Enables Expansion of Disease-Specific Regulatory T Cells in an Animal Model of Multiple Sclerosis.
Disease activity of autoimmune disorders such as multiple sclerosis and its mouse model experimental autoimmune encephalomyelitis (EAE) is temporarily suppressed by pregnancy. However, whether disease amelioration is due to nonspecific immunomodulation or mediated by Ag-specific regulation of disease-causing conventional T cells (Tcon) and immunosuppressive regulatory T cells (Tregs) remains elusive. In the current study, we systematically analyzed changes of the TCRβ repertoire driven by EAE and pregnancy using TCR sequencing. We demonstrate that EAE, but not pregnancy, robustly increased TCR repertoire clonality in both peripheral Tcon and Treg. Notably, pregnancy was required for the expansion of Treg harboring the dominant EAE-associated TRBV13-2 chain and increased the frequency of EAE-associated clonotypes within the Treg compartment. Our findings indicate that pregnancy supports the expansion of Treg clonotypes that are equipped to recognize EAE-associated Ags. These Treg are thereby particularly suited to control corresponding encephalitogenic Tcon responses and likely contribute to pregnancy-associated protection in autoimmunity.


Identifying natural images from human brain activity.
A challenging goal in neuroscience is to be able to read out, or decode, mental content from brain activity. Recent functional magnetic resonance imaging (fMRI) studies have decoded orientation, position and object category from activity in visual cortex. However, these studies typically used relatively simple stimuli (for example, gratings) or images drawn from fixed categories (for example, faces, houses), and decoding was based on previous measurements of brain activity evoked by those same stimuli or categories. To overcome these limitations, here we develop a decoding method based on quantitative receptive-field models that characterize the relationship between visual stimuli and fMRI activity in early visual areas. These models describe the tuning of individual voxels for space, orientation and spatial frequency, and are estimated directly from responses evoked by natural images. We show that these receptive-field models make it possible to identify, from a large set of completely novel natural images, which specific image was seen by an observer. Identification is not a mere consequence of the retinotopic organization of visual areas; simpler receptive-field models that describe only spatial tuning yield much poorer identification performance. Our results suggest that it may soon be possible to reconstruct a picture of a person's visual experience from measurements of brain activity alone.


Targeted deletion of AP-2alpha leads to disruption in corneal epithelial cell integrity and defects in the corneal stroma.
The present study was undertaken to create a conditional knockout of AP-2alpha in the corneal epithelium. A line of mice expressing Cre-recombinase specifically in the early lens placode was crossed with mice in which the AP-2alpha allele is flanked by two loxP sites. The resultant Le-AP-2alpha mutants exhibited a targeted deletion of AP-2alpha in lens placode derivatives, including the differentiating corneal epithelium. The Le-AP-2alpha mutant mice were viable and had a normal lifespan. The adult corneal epithelium exhibited a variation in the number of stratified epithelial layers, ranging from 2 to 10 cell layers. A substantial decrease in expression of the cell-cell adhesion molecule, E-cadherin, was observed in all layers of the Le-AP-2alpha mutant corneal epithelium. The basement membrane, or Bowman's layer, was thinner in the mutant cornea and in many regions was discontinuous. These defects corresponded with altered distribution of laminin and entactin, and to a lesser degree, type IV collagen. The Le-AP-2alpha mutant cornea also exhibited stromal defects, including disrupted organization of the collagen lamellae and accumulation of fibroblasts beneath the epithelium that showed increased immunoreactivity for proliferating cell nuclear antigen (PCNA), alpha-smooth muscle actin (alpha-SMA), p-Smad2, and TGF-beta2. In the absence of AP-2alpha, the corneal epithelium exhibits altered cell adhesion and integrity and defects in its underlying basement membrane. These defects likely caused the alterations in the corneal stroma.


Research priorities for the management of broken bones of the upper limb in people over 50: a UK priority setting partnership with the James Lind Alliance.
To determine research priorities for the management of broken bones of the upper limb in people over 50, which represent the shared priorities of patients, their families, carers and healthcare professionals. A national (UK) research priority setting partnership. People aged 50 and over who have experienced a fracture of the upper limb, carers involved in their care, family and friends of patients, healthcare professionals involved in the treatment of these patients. Using a multiphase methodology in partnership with the James Lind Alliance over 15 months (September 2017 to December 2018), a national scoping survey asked respondents to submit their research uncertainties. These were amalgamated into a smaller number of research questions. The existing evidence was searched to ensure that the questions had not already been answered. A second national survey asked respondents to prioritise the research questions. A final shortlist of 25 questions was taken to a multi-stakeholder workshop where a consensus was reached on the top 10 priorities. There were 1898 original uncertainties submitted by 328 respondents to the first survey. These original uncertainties were refined into 51 research questions of which 50 were judged to be true uncertainties following a review of the research evidence. There were 209 respondents to the second (interim prioritisation) survey. The top 10 priorities encompass a broad range of uncertainties in management and rehabilitation of upper limb fractures. The top 10 UK research priorities highlight uncertainties in how we assess outcomes, provide information, achieve pain control, rationalise surgical intervention, optimise rehabilitation and provide psychological support. The breadth of these research areas highlights the value of this methodology. This work should help to steer research in this area for the next 5-10 years and the challenge for researchers now is to refine and deliver answers to these research priorities.


Tissue microarrays: one size does not fit all.
Although tissue microarrays (TMAs) are commonly employed in clinical and basic-science research, there are no guidelines for evaluating the appropriateness of a TMA for a given biomarker and tumor type. Furthermore, TMA performance across multiple biomarkers has not been systematically explored. A simulated TMA with between 1 and 10 cores was designed to study tumor expression of 6 biomarkers with varied expression patterns (B7-H1, B7-H3, survivin, Ki-67, CAIX, and IMP3) using 100 patients with clear cell renal cell carcinoma (RCC). We evaluated agreement between whole tissue section and TMA immunohistochemical biomarker quantification to assess how many TMA cores are necessary to adequately represent RCC whole tissue section expression. Additionally, we evaluated associations of whole tissue section and TMA expression with RCC-specific death. The number of simulated TMA cores necessary to adequately represent whole tissue section quantification is biomarker specific. Although 2-3 cores appeared adequate for B7-H3, Ki-67, CAIX, and IMP3, even as many as 10 cores resulted in poor agreement for B7-H1 and survivin compared to RCC whole tissue sections. While whole tissue section B7-H1 was significantly associated with RCC-specific death, no significant associations were detected using as many as 10 TMA cores, suggesting that TMAs can result in false-negative findings if the TMA is not optimally designed. Prior to TMA analysis, the number of TMA cores necessary to accurately represent biomarker expression on whole tissue sections should be established as there is not a one-size-fits-all TMA. We illustrate the use of a simulated TMA as a cost-effective tool for this purpose.