Molecular Basis of Binding between Middle East Respiratory Syndrome Coronavirus and CD26 from Seven Bat Species.
Continued reports of Middle East respiratory syndrome coronavirus (MERS-CoV) infecting humans have occurred since the identification of this virus in 2012. MERS-CoV is prone to cause endemic disease in the Middle East, with several dozen spillover infections to other continents. It is hypothesized that MERS-CoV originated from bat coronaviruses and that dromedary camels are its natural reservoir. Although gene segments identical to MERS-CoV were sequenced from certain species of bats and one species experimentally shed the virus, it is still unknown whether other bats can transmit the virus. Here, at the molecular level, we found that all purified bat CD26s (bCD26s) from a diverse range of species interact with the receptor binding domain (RBD) of MERS-CoV, with equilibrium dissociation constant values ranging from several to hundreds at the micromolar level. Moreover, all bCD26s expressed in this study mediated the entry of pseudotyped MERS-CoV to receptor-expressing cells, indicating the broad potential engagement of bCD26s as MERS-CoV receptors. Further structural analysis indicated that in the bat receptor, compared to the human receptor, substitutions of key residues and their adjacent amino acids leads to decreased binding affinity to the MERS-RBD. These results add more evidence to the existing belief that bats are the original source of MERS-CoV and suggest that bCD26s in many species can mediate the entry of the virus, which has significant implications for the surveillance and control of MERS-CoV infection.IMPORTANCE In this study, we found that bat CD26s (bCD26s) from different species exhibit large diversities, especially in the region responsible for binding to the receptor binding domain (RBD) of Middle East respiratory syndrome coronavirus (MERS-CoV). However, they maintain the interaction with MERS-RBD at varied affinities and support the entry of pseudotyped MERS-CoV. These bat receptors polymorphisms seem to confer evolutionary pressure for the adaptation of CD26-binding virus, such as the ancestor of MERS-CoV, and led to the generation of diversified CD26-engaging CoV strains. Thus, our data add more evidence to support that bats are the reservoir of MERS-CoV and similar viruses, as well as further emphasize the necessity to survey MERS-CoV and other CoVs among bats.


Tissue culture and animal toxicity studies of antineoplaston A5.
Antineoplaston A5 is a mixture of small to medium size peptides and amino acid derivatives isolated from human urine. This report describes growth inhibition by Antineoplaston A5 of human cell carcinoma line HBL-100 and human leukaemic cell line Jurkat, and acute and chronic toxicity studies in mice. The inhibitory effect of Antineoplaston A5 on HBL-100 cells was tested at concentrations varying from 0.125 mg/ml to 0.5 mg/ml and at exposure durations varying from 24 to 96 h. The tests on Jurkat cells were carried out at concentrations varying from 0.1 mg/ml to 0.8 mg/ml and exposure durations varying from 24 to 120 h. The results from HBL-100 cells, shown as percentage of survival, were as follows: at 0.125 mg/ml 58%; at 0.25 mg/ml 14%; at 0.5 mg/ml 0. Exposure of HBL-100 cells to 0.5 mg/ml of Antineoplaston A5 for 24, 48, 72 and 96 h reduced survival to 55%, 10%, 4% and 0 of the control respectively. The results for suspension culture of Jurkat were as follows: The control cells showed a 7.7-fold increase in cell number during the incubation period, while the cells treated with 0.8 mg/ml of Antineoplaston A5 showed a steady decrease in cell number. At the end of 120 hours' incubation, only 1% of the cells was still viable. The growth inhibition patterns for dosages ranging from 0.4 mg/ml to 0.1 mg/ml were unique; instead of decreasing, the cell numbers in these flasks kept increasing, although at a slower pace during the initial 72 h.(ABSTRACT TRUNCATED AT 250 WORDS)


Variations between the nucleotide sequences of Tn1, Tn2, and Tn3 and expression of beta-lactamase in Pseudomonas aeruginosa and Escherichia coli.
Using S1 nuclease assays, we located the sites of initiation of transcription of the beta-lactamase gene on Tn1 and Tn2. Transcription in Tn2, like that in Tn3, occurred from the P3 promoter, whereas transcription in Tn1 was initiated by two stronger and overlapping promoters, Pa and Pb. The nucleotide sequences of Tn1 and Tn2 were determined over a 1,195-base-pair segment constituting most of the sequences of the tnpR and bla genes and the intervening region. There were six base-pair differences between Tn1 and Tn3. One in the bla regulatory region accounted for the presence of the Pa and Pb promoters, and another in the bla structural gene is consistent with the isoelectric focusing difference found between the Tn1 and Tn3 enzymes. In contrast, there were 24 base-pair differences between Tn2 and Tn3, most of them clustered in one segment of the tnpR gene.


Enhanced diurnal variation of blood pressure in the renal hypertensive rat: effect of angiotensin II suppression.
1. Chronic two-kidney, one-clip hypertensive rats were infused with captopril for 5 days and the daily variability of blood pressure compared with that for both hypertensive rats infused with glucose and normotensive animals. 2. Blood pressure was measured continuously using a computer data collecting system. 3. Normotensive animals showed a stable level of arterial pressure throughout each 24 h period with troughs occurring when they slept during the daytime. 4. Hypertensive animals given glucose had an enhanced diurnal rhythm of blood pressure compared with normotensive rats, with peaks occurring during periods of activity at night as well as troughs when they slept during the day. 5. Hypertensive rats given captopril retained this enhanced pressure variability, in spite of the fact that blood pressure was significantly lower and angiotensin II was suppressed. 6. These results suggest that angiotensin II is not involved in the increased blood pressure variability of renal hypertension and that some other irreversible mechanism is responsible.


Stiffness Is the Cardinal Symptom of Inflammatory Musculoskeletal Diseases, Yet Still Variably Measured: Report from the OMERACT 2016 Stiffness Special Interest Group.
The objectives of the Outcome Measures in Rheumatology (OMERACT) Stiffness special interest group (SIG) are to characterize stiffness as an outcome in rheumatic disease and to identify and validate a stiffness patient-reported outcome (PRO) in rheumatology. At OMERACT 2016, international groups presented and discussed results of several concurrent research projects on stiffness: a literature review of rheumatoid arthritis (RA) stiffness PRO measures, a qualitative investigation into the RA and polymyalgia rheumatica patient perspective of stiffness, data-driven stiffness conceptual model development, development and testing of an RA stiffness PRO measure, and a quantitative work testing stiffness items in patients with RA and psoriatic arthritis. The literature review identified 52 individual stiffness PRO measures assessing morning or early morning stiffness severity/intensity or duration. Items were heterogeneous, had little or inconsistent psychometric property evidence, and did not appear to have been developed according to the PRO development guidelines. A poor match between current stiffness PRO and the conceptual model identifying the RA patient experience of stiffness was identified, highlighting a major flaw in PRO selection according to the OMERACT filter 2.0. Discussions within the Stiffness SIG highlighted the importance of further research on stiffness and defined a research agenda.


Registered nurses' application of evidence-based practice: a national survey.
Evidence-based practice (EBP) is a worldwide approach to improving health care. There is, however, a shortage of studies examining whether or not newly graduated health care professionals are actually applying EBP in their daily work. To examine the application of EBP in clinical practice by registered nurses (RNs) 2 years post graduation and to explore whether the application of EBP differed with regard to the clinical settings where RNs were working. A cross-sectional design using a national sample. Data were collected in 2007 from 987 RNs (response rate 76%). Six items measuring respondents' self-reported extent of applying EBP were used. Results Of the 987 RNs, 19% formulated questions and performed searches in data bases, 56% used other information sources, 31% appraised the literature, 30% participated in practice development and 34% participated in evaluating clinical practice. A greater proportion of the RNs working in elder care applied EBP compared with the RNs working in hospitals, psychiatric care and primary care. The RNs applied the components of EBP to a rather low extent 2 years post graduation despite EBP being an important objective in Swedish health care and educational programmes since the 1990s. These findings support other studies reporting the implementation of EBP in organizations as a complex and often slow process. The differences in the RNs extent of applying EBP in relation to their workplace indicate that contextual factors and the role of the RN in the organization are of importance for getting EBP into practice.


Characterization of coupled nonlinear spatiospectral phase following an ultrafast self-focusing interaction.
We use collinear spatially resolved spectral interferometery to characterize the nonlinear phase changes experienced by an intense ultrashort pulse propagating in glass. The measurement yields the spectrally dependent wavefront, allowing us to measure the spatial and chromatic aberrations of the nonlinearly induced lens. For these conditions, we find that while the shape of the spatial wavefront follows the beam profile as expected, the spectral dependence of the lensing power is determined by the self-phase modulation. The simultaneous measurement of the nonlinear spatiospectral phase demonstrates how the nonlinear spectral phase is coupled to self-focusing.


Heating and destruction of metallic particles exposed to intense laser radiation.
The heating of a laser-irradiated solid aluminum particle to boiling or to temperatures that exceed boiling is analyzed theoretically and numerically by solution of the heat-transport equation. Two different criteria of particle destruction are considered. The temperature distributions inside the particles depending on the intensity values and particle sizes are presented. It is shown that at the start of heating the contribution of heat exchange plays the dominant role, but as the boiling point is approached the contribution of vaporization plays the main role.


IL-35 promotes CD4+Foxp3+ Tregs and inhibits atherosclerosis via maintaining CCR5-amplified Treg-suppressive mechanisms.
Tregs play vital roles in suppressing atherogenesis. Pathological conditions reshape Tregs and increase Treg-weakening plasticity. It remains unclear how Tregs preserve their function and how Tregs switch into alternative phenotypes in the environment of atherosclerosis. In this study, we observed a great induction of CD4+Foxp3+ Tregs in the spleen and aorta of ApoE-/- mice, accompanied by a significant increase of plasma IL-35 levels. To determine if IL-35 devotes its role in the rise of Tregs, we generated IL-35 subunit P35-deficient (IL-35P35-deficient) mice on an ApoE-/- background and found Treg reduction in the spleen and aorta compared with ApoE-/- controls. In addition, our RNA sequencing data show the elevation of a set of chemokine receptor transcripts in the ApoE-/- Tregs, and we have validated higher CCR5 expression in ApoE-/- Tregs in the presence of IL-35 than in the absence of IL-35. Furthermore, we observed that CCR5+ Tregs in ApoE-/- have lower Treg-weakening AKT-mTOR signaling, higher expression of inhibitory checkpoint receptors TIGIT and PD-1, and higher expression of IL-10 compared with WT CCR5+ Tregs. In conclusion, IL-35 counteracts hyperlipidemia in maintaining Treg-suppressive function by increasing 3 CCR5-amplified mechanisms, including Treg migration, inhibition of Treg weakening AKT-mTOR signaling, and promotion of TIGIT and PD-1 signaling.


Sphingobium terrigena sp. nov., isolated from gasoline-contaminated soil.
A Gram-stain-negative, strictly aerobic bacterial strain, designated EO9T, was isolated from gasoline-contaminated soil in the Republic of Korea. Cells were non-motile short rods showing catalase- and oxidase-positive reactions. Growth was observed at 10-37 °C (optimum, 30 °C), at pH 6.0-9.0 (pH 6.5) and in the presence of 0-0.5 % (w/v) NaCl (0 %). Ubiquinone-10 (Q-10) and spermidine were identified as the predominant respiratory quinone and polyamine, respectively. Summed feature 8 (comprising C18:1ω7c/C18:1ω6c), summed feature 3 (comprising C16:1ω7c/C16:1ω6c), C16:0 and C14:0 2-OH were identified as major cellular fatty acids. The major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, sphingoglycolipid, phosphatidylglycerol and an unidentified phospholipid. The G+C content of the genomic DNA was 62.8 mol%. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain EO9T formed a tight phylogenetic lineage with Sphingobium xenophagum NBRC 107872T and Sphingobium hydrophobicum C1T within the genus Sphingobium. Strain EO9T was most closely related to S. xenophagum NBRC 107872T (97.2 %) and S. hydrophobicum C1T (97.2 %), but DNA-DNA relatedness levels between strain EO9T and the type strains of S. xenophagum and S. hydrophobicum were 37.1 and 36.8 % , respectively. Based on its phenotypic, chemotaxonomic and molecular features, strain EO9T clearly represents a novel species of the genus Sphingobium, for which the name Sphingobium terrigena sp. nov. is proposed. The type strain is EO9T (=KACC 19523T=JCM 32762T).


Impact of food and predation on the snowshoe hare cycle.
Snowshoe hare populations in the boreal forests of North America go through 10-year cycles. Supplemental food and mammalian predator abundance were manipulated in a factorial design on 1-square-kilometer areas for 8 years in the Yukon. Two blocks of forest were fertilized to test for nutrient effects. Predator exclosure doubled and food addition tripled hare density during the cyclic peak and decline. Predator exclosure combined with food addition increased density 11-fold. Added nutrients increased plant growth but not hare density. Food and predation together had a more than additive effect, which suggests that a three-trophic-level interaction generates hare cycles.


Functional analysis of a type 1 parathyroid hormone receptor intracellular tail mutant [KRK(484-6)AAA]: effects on second messenger generation and cellular targeting.
The parathyroid hormone receptor type 1 (PTHR1) is activated by parathyroid hormone (PTH) and PTH-related protein (PTHrP) and primarily signals via intracellular pathways involving adenylyl cyclase and phospholipase C. The intracellular tail domain of the PTHR1 contributes to G protein subunit coupling that is important for second messenger signalling. In addition, the intracellular domain has a potential nuclear localization sequence (NLS) that, if functional, could point to an intracrine role for the receptor. In the present study, we have utilized 2 sets of constructs that employ either a [KRK(484-486)AAA](3Ala) mutation in the putative NLS or the non-mutant counterpart and included (a) the full-length rat PTHR1 with FLAG and c-myc epitope tags at the N-terminus and C-terminus, respectively (designated as PTHR1(3Ala)-TAG and PTHR1-TAG); and (b) only the putative NLS-containing intracellular domain (471-488), with green fluorescent protein (GFP) fused to the C-terminus (designated as GFP-(3Ala)471-488 or GFP-471-488). Porcine kidney LLC-PK1 cells stably expressing the PTHR1(3Ala)-TAG exhibited reduced signalling via both cAMP and cytosolic calcium transients in spite of greater cell surface expression relative to cells expressing PTHR1-TAG. We also examined the ability of the intracellular tail to influence the cellular localization of a heterologous protein. LLC-PK1 cells transiently transfected with GFP-471-488, exhibited increased fluorescence within the nucleus, relative to cells transfected with GFP alone that was not observed when cells were transiently transfected with the mutated construct, GFP-(3Ala)471-488. However, LLC-PK1 cells transiently transfected with either the full-length PTHR1-TAG or the PTHR1(3Ala)-TAG constructs did not exhibit nuclear localization of these receptors. Moreover, mouse osteoblast-like cells (MC3T3-E1) transiently expressing PTHR1-TAG also failed to demonstrate nuclear localization, although both full-length PTHR1 constructs exhibited plasma membrane immunofluorescence in both cell lines. Thus, the 484-486 sequence is critical for the full signalling responsiveness of the intact PTHR1, but the putative nuclear localization signal may not function as such within the intact receptor.


No evidence for the association between a polymorphism in the PCLO depression candidate gene with memory bias in remitted depressed patients and healthy individuals.
The PCLO rs2522833 candidate polymorphism for depression has been associated to monoaminergic neurotransmission. In healthy and currently depressed individuals, the polymorphism has been found to affect activation of brain areas during memory processing, but no direct association of PCLO with memory bias was found. We hypothesized that the absence of this association might have been obscured by current depressive symptoms or genetically driven individual differences in reactivity to stressful events. Experiencing stressful childhood events fosters dysfunctional assumptions that are related to cognitive biases, and may modulate the predisposition for depression via epigenetic effects. The association between PCLO and memory bias, as well as interaction between PCLO and childhood events was studied in patients remitted from depression (N = 299), as well as a sample of healthy individuals (N = 157). The participants performed an emotional verbal memory task after a sad mood induction. Childhood trauma and adversity were measured with a questionnaire. The Genotype main effect, and Genotype by Childhood Events interaction were analyzed for memory bias in both samples. PCLO risk allele carrying remitted depressed patients did not show more negatively biased memory than non-risk allele carriers, not even patients with stressful childhood events. A similar pattern of results was found in healthy individuals. Memory bias may not be strongly associated with the PCLO rs2522833 polymorphism. We did not find any support for the PCLO-childhood events interaction, but the power of our study was insufficient to exclude this possibility.


Pornography Use Prevalence and Associated Factors in Arab Countries: A Multinational Cross-Sectional Study of 15,027 Individuals.
There has been a tangible increase in pornography consumption during the past decade, with the absence of a large-scale study of Arab countries. The present study aimed to assess the prevalence of pornography viewing and its associated risk factors in Arab countries. A large cross-sectional online survey was carried out recruiting participants without restrictions on the age, socioeconomic level, job, or educational level. Multivariable logistic regression analysis was performed to identify possible risk factors for viewing porn and results were expressed as odds ratios (ORs) and 95% confidence interval (95% CI). Main outcomes were pornography viewing (first exposure and frequency), the perceptions of this act, use of spare time, physical activity (exercising), and frequency of psychiatrist visits. The final number of participants included in the study was 15027 participants with a mean age ± standard deviation of 23.82 years ± 24.99. Most of the participants were men (84.56%), living with parents (81.71%), and 60.51% university graduates. There were statistically significant differences (P < .001) in the attitude and practice of men compared with women throughout all tested variables. Frequent pornography viewing was associated with male gender (OR [95% CI] = 7.08 [6.43 to 7.81]; P < .001) and age group ≤15 years (OR [95% CI] = 1.33 [1.01 to 1.75]; P = .044). By contrast, higher education was inversely associated with viewing rates reaching the lowest level in PhD awardees (OR [95% CI] = 0.36 [0.26 to 0.51]; P = .003). It was also noted that regular exercising (OR [95% CI] = 0.66 [0.58 to 0.74]; P < .001) was associated with a reduction in pornography viewing rates. Young age, male gender, and lower educational level are all predictors for higher pornography viewing and should be considered when designing public health intervention in a related context. This is the first large-scale multi-national survey to be conducted in Arab countries to investigate pornography viewing. The main limitations were the cross-sectional design (cannot indicate causality) and the self-report nature (liable to social desirability and recall bias). Pornography viewing is common in Arab countries and associated with some personal and behavioral factors. Eljawad MA, Se'eda H, Ghozy S, et al. Pornography Use Prevalence and Associated Factors in Arab Countries: A Multinational Cross-Sectional Study of 15,027 Individuals. J Sex Med 2021;18:539-548.


AXL Inactivation Inhibits Mesothelioma Growth and Migration via Regulation of p53 Expression.
Malignant mesothelioma is a locally aggressive and highly lethal neoplasm. Dysregulation and activation of Gas6/AXL tyrosine kinase signaling are associated with mesothelioma progression, but the mechanisms of these AXL tumorigenic roles are poorly understood. p53 mutants in lung carcinoma upregulate AXL expression by binding and acetylating the AXL promoter. Although TP53 mutations are uncommon in mesothelioma, we hypothesized that these tumors might have alternative feedback mechanisms between AXL and p53. In the current report, we investigated AXL regulation of TP53 transcription, expression, and biological function in mesothelioma. AXL expression was stronger in mesothelioma than most of the other tumor types from the TCGA gene expression profile dataset. AXL knockdown by shRNA induced wild-type and mutant p53 expression in mesothelioma cell lines, suggesting that AXL pro-tumorigenic roles result in part from the suppression of p53 function. Likewise, induced AXL inhibited expression of wild type p53 in COS-7 cells and 293T cells. Immunofluorescence staining showed nuclear colocalization of AXL and p53; however, association of AXL and p53 was not demonstrated in immunoprecipitation complexes. The AXL effects on p53 expression resulted from the inhibition of TP53 transcription, as demonstrated by qRT-PCR after AXL silencing and TP53 promotor dual luciferase activity assays. Chromatin immunoprecipitation-qPCR and sequencing showed that AXL bound to the initial 600 bp sequence at the 5' end of the TP53 promoter. AXL inhibition (shRNA or R428) reduced mesothelioma cell viability, migration, and invasion, whereas TP53 shRNA knockdown attenuated antiproliferative, migration, and invasive effects of AXL silencing or AXL inactivation in these cells. These studies demonstrate a novel feedback regulation loop between AXL and p53, and provide a rationale for mesothelioma therapies targeting AXL/p53 signaling.


Prevalence of pfhrp2 and pfhrp3 gene deletions in Puerto Lempira, Honduras.
Recent studies have demonstrated the deletion of the histidine-rich protein 2 (PfHRP2) gene (pfhrp2) in field isolates of Plasmodium falciparum, which could result in false negative test results when PfHRP2-based rapid diagnostic tests (RDTs) are used for malaria diagnosis. Although primary diagnosis of malaria in Honduras is determined based on microscopy, RDTs may be useful in remote areas. In this study, it was investigated whether there are deletions of the pfhrp2, pfhrp3 and their respective flanking genes in 68 P. falciparum parasite isolates collected from the city of Puerto Lempira, Honduras. In addition, further investigation considered the possible correlation between parasite population structure and the distribution of these gene deletions by genotyping seven neutral microsatellites. Sixty-eight samples used in this study, which were obtained from a previous chloroquine efficacy study, were utilized in the analysis. All samples were genotyped for pfhrp2, pfhrp3 and flanking genes by PCR. The samples were then genotyped for seven neutral microsatellites in order to determine the parasite population structure in Puerto Lempira at the time of sample collection. It was found that all samples were positive for pfhrp2 and its flanking genes on chromosome 8. However, only 50% of the samples were positive for pfhrp3 and its neighboring genes while the rest were either pfhrp3-negative only or had deleted a combination of pfhrp3 and its neighbouring genes on chromosome 13. Population structure analysis predicted that there are at least two distinct parasite population clusters in this sample population. It was also determined that a greater proportion of parasites with pfhrp3-(and flanking gene) deletions belonged to one cluster compared to the other. The findings indicate that the P. falciparum parasite population in the municipality of Puerto Lempira maintains the pfhrp2 gene and that PfHRP2-based RDTs could be considered for use in this region; however continued monitoring of parasite population will be useful to detect any parasites with deletions of pfhrp2.


Allosteric nanobodies reveal the dynamic range and diverse mechanisms of G-protein-coupled receptor activation.
G-protein-coupled receptors (GPCRs) modulate many physiological processes by transducing a variety of extracellular cues into intracellular responses. Ligand binding to an extracellular orthosteric pocket propagates conformational change to the receptor cytosolic region to promote binding and activation of downstream signalling effectors such as G proteins and β-arrestins. It is well known that different agonists can share the same binding pocket but evoke unique receptor conformations leading to a wide range of downstream responses ('efficacy'). Furthermore, increasing biophysical evidence, primarily using the β2-adrenergic receptor (β2AR) as a model system, supports the existence of multiple active and inactive conformational states. However, how agonists with varying efficacy modulate these receptor states to initiate cellular responses is not well understood. Here we report stabilization of two distinct β2AR conformations using single domain camelid antibodies (nanobodies)—a previously described positive allosteric nanobody (Nb80) and a newly identified negative allosteric nanobody (Nb60). We show that Nb60 stabilizes a previously unappreciated low-affinity receptor state which corresponds to one of two inactive receptor conformations as delineated by X-ray crystallography and NMR spectroscopy. We find that the agonist isoprenaline has a 15,000-fold higher affinity for β2AR in the presence of Nb80 compared to the affinity of isoprenaline for β2AR in the presence of Nb60, highlighting the full allosteric range of a GPCR. Assessing the binding of 17 ligands of varying efficacy to the β2AR in the absence and presence of Nb60 or Nb80 reveals large ligand-specific effects that can only be explained using an allosteric model which assumes equilibrium amongst at least three receptor states. Agonists generally exert efficacy by stabilizing the active Nb80-stabilized receptor state (R80). In contrast, for a number of partial agonists, both stabilization of R80 and destabilization of the inactive, Nb60-bound state (R60) contribute to their ability to modulate receptor activation. These data demonstrate that ligands can initiate a wide range of cellular responses by differentially stabilizing multiple receptor states.


Amine scrubbing for CO2 capture.
Amine scrubbing has been used to separate carbon dioxide (CO2) from natural gas and hydrogen since 1930. It is a robust technology and is ready to be tested and used on a larger scale for CO2 capture from coal-fired power plants. The minimum work requirement to separate CO2 from coal-fired flue gas and compress CO2 to 150 bar is 0.11 megawatt-hours per metric ton of CO2. Process and solvent improvements should reduce the energy consumption to 0.2 megawatt-hour per ton of CO2. Other advanced technologies will not provide energy-efficient or timely solutions to CO2 emission from conventional coal-fired power plants.


A Comparison of DNA Mutation and Copy Number Profiles of Primary Breast Cancers and Paired Brain Metastases for Identifying Clinically Relevant Genetic Alterations in Brain Metastases.
Improving the systemic treatment of brain metastases (BM) in primary breast cancer (PBC) is impaired by the lack of genomic characterization of BM. To estimate the concordance of DNA copy-number-alterations (CNAs), mutations, and actionable genetic alterations (AGAs) between paired samples, we performed whole-genome array-comparative-genomic-hybridization, and targeted-next-generation-sequencing on 14 clinical PBC-BM pairs. We found more CNAs, more mutations, and higher tumor mutational burden, and more AGAs in BM than in PBC; 92% of the pairs harbored at least one AGA in the BM not observed in the paired PBC. This concerned various therapeutic classes, including tyrosine-kinase-receptor-inhibitors, phosphatidylinositol 3-kinase/AKT/ mammalian Target of Rapamycin (PI3K/AKT/MTOR)-inhibitors, poly ADP ribose polymerase (PARP)-inhibitors, or cyclin-dependent kinase (CDK)-inhibitors. With regards to the PARP-inhibitors, the homologous recombination defect score was positive in 79% of BM, compared to 43% of PBC, discordant in 7 out of 14 pairs, and positive in the BM in 5 out of 14 cases. CDK-inhibitors were associated with the largest percentage of discordant AGA appearing in the BM. When considering the AGA with the highest clinical-evidence level, for each sample, 50% of the pairs harbored an AGA in the BM not detected or not retained from the analysis of the paired PBC. Thus, the profiling of BM provided a more reliable opportunity, than that of PBC, for diagnostic decision-making based on genomic analysis. Patients with BM deserve an investigation of several targeted therapies.


Aqueous-solution-processable metal oxides for high-performance organic and perovskite solar cells.
Poly(3,4-ethylene dioxythiophene):poly(4-styrenesulfonate) (PEDOT:PSS) is a widely utilized hole-transporting material (HTM) in planar photovoltaic devices, such as organic solar cells (OSCs) and perovskite solar cells (PSCs). However, the hygroscopic nature of PEDOT:PSS aqueous dispersions may restrict their future application. Therefore, it is necessary to develop other effective and stable HTMs to achieve high-performance photovoltaic devices. Herein, we demonstrate a facile route to deposit solution-processed MoO3, GeO2, V2O5 and CrO3 thin films as hole-transporting layers by directly dissolving their commercial powders in deionized water. Among these, the solution-processed V2O5 (sV2O5) film exhibited the highest work function of 5.2 eV, and the best hydrophobicity, with a contact angle of 77.2°. The sV2O5-based OSCs and PSCs presented power conversion efficiencies (PCEs) of 8.36% and 14.13%, respectively. Notably, the PEDOT:PSS V2O5 composite HTM based device obtained a maximum PCE of 18.03% with a Voc exceeding 1.0 V. These aqueous-solution-processed HTMs have potential applications in green and low-cost photovoltaic devices by virtue of their simple and ecofriendly preparations.


Worsening renal function and outcome in heart failure patients with preserved ejection fraction and the impact of angiotensin receptor blocker treatment.
Worsening renal function (WRF) associated with renin-angiotensin-aldosterone system (RAAS) inhibition does not confer excess risk in heart failure patients with reduced ejection fraction (HFrEF). The goal of this study was to investigate the relationship between WRF and outcomes in heart failure patients with preserved ejection fraction (HFpEF) and the interaction with RAAS blockade. In 3,595 patients included in the I-PRESERVE (Irbesartan in Heart Failure With Preserved Ejection Fraction) trial, change in estimated glomerular filtration rate (eGFR) and development of WRF after initiation of irbesartan or placebo were examined. We examined the association between WRF and the first occurrence of cardiovascular death or heart failure hospitalization (primary outcome in this analysis) and the interaction with randomized treatment. Estimated GFR decreased early with irbesartan treatment and remained significantly lower than in the placebo group. WRF developed in 229 (6.4%) patients and occurred more frequently with irbesartan treatment (8% vs. 4%). Overall, WRF was associated with an increased risk of the primary outcome (adjusted hazard ratio [HR]: 1.43; 95% confidence interval [CI]: 1.10 to 1.85; p = 0.008). Although the risk related to WRF was greater in the irbesartan group (HR: 1.66; 95% CI: 1.21 to 2.28; p = 0.002) than with placebo (HR: 1.09; 95% CI: 0.66 to 1.79; p = 0.73), the interaction between treatment and WRF on outcome was not significant in an adjusted analysis. The incidence of WRF in HFpEF was similar to that previously reported in HFrEF but more frequent with irbesartan than with placebo. WRF after initiation of irbesartan treatment in HFpEF was associated with excess risk, in contrast to WRF occurring with RAAS blockade in HFrEF.


A descriptive epidemiology of netball injuries during competition: a five year study.
To investigate the relation between injury profiles - including assessment, treatment, severity, and the perceived reason for the injury - and participation in competitive netball. Between 1985 and 1989, approximately 11,228 netball players participated in a 14 week netball competition held at the major competitive centre in Western Australia; 608 netball players presented to the first aid room with an injury and were assessed and treated by the same physiotherapist and St John First Aid officer. The overall incidence rate during competition was 5.4%, with more injuries occurring in the A grade level of play. Ankle injuries (84%) were most frequent, with 67% of these injuries diagnosed as lateral ligament sprains while a further 10% of players who suffered this classification of injury sustained a fracture to the ankle or foot. Few injuries occurred at the knee joint (8.3%) and only 1.8% of these injuries were diagnosed as an injury to the anterior cruciate ligament. The direct probability of a netball players' risk estimate was 0.054 per person per match which implied that netball was a relatively safe game. Injuries sustained during practice were not included in this study. Netball is a relatively safe game though the potential for injury increases with the level of competition.


Sleep Promotes Phonological Learning in Children Across Language and Autism Spectra.
Purpose Establishing stable and flexible phonological representations is a key component of language development and one which is thought to vary across children with neurodevelopmental disorders affecting language acquisition. Sleep is understood to support the learning and generalization of new phonological mappings in adults, but this remains to be examined in children. This study therefore explored the time course of phonological learning in childhood and how it varies by structural language and autism symptomatology. Method Seventy-seven 7- to 13-year-old children, 30 with high autism symptomatology, were included in the study; structural language ability varied across the sample. Children learned new phonological mappings based on synthesized speech tokens in the morning; performance was then charted via repetition (without feedback) over 24 hr and followed up 4 weeks later. On the night following learning, children's sleep was monitored with polysomnography. Results A period of sleep but not wake was associated with improvement on the phonological learning task in childhood. Sleep was associated with improved performance for both trained items and novel items. Structural language ability predicted overall task performance, though language ability did not predict degree of change from one session to the next. By contrast, autism symptomatology did not explain task performance. With respect to sleep architecture, rapid eye movement features were associated with greater phonological generalization. Conclusions Children's sleep was associated with improvement in performance on both trained and novel items. Phonological generalization was associated with brain activity during rapid eye movement sleep. This study furthers our understanding of individual differences in the acquisition of new phonological mappings and the role of sleep in this process over childhood. Supplemental Material https://doi.org/10.23641/asha.11126732.


Effects of mechanical and thermal aging on microleakage of different fissure sealants.
The purpose of this study was to examine the microleakage of three different fissure sealants after they were aged by mechanical loading and thermocycling in vitro. To this end, a bonding agent (Prime & Bond NT) and three different fissure sealants (Clinpro, Helioseal F, Teethmate F1) were used, whereby microleakage was evaluated using a dye penetration method after mechanical loading and/or thermocycling. Sealant-treated teeth were allocated into four groups: mechanical loading (50,000 times), thermocycling (10,000 times), mechanical loading (50,000 times) + thermocycling (10,000 times), and one control group. For each fissure sealant, both experimental and control groups showed statistically significant differences in average microleakage score (p < 0.05). Further, for each fissure sealant, the highest average microleakage score was obtained in mechanical loading + thermocycling group. When comparison was done for each aging method, the average microleakage scores showed statistically significant differences among the three fissure sealants (p < 0.05). Based on the results of this study, it was also concluded that it is necessary to develop reliable in vitro test methods for dental materials.


Evaluating the impact of the DREAMS partnership to reduce HIV incidence among adolescent girls and young women in four settings: a study protocol.
HIV risk remains unacceptably high among adolescent girls and young women (AGYW) in southern and eastern Africa, reflecting structural and social inequities that drive new infections. In 2015, PEPFAR (the United States President's Emergency Plan for AIDS Relief) with private-sector partners launched the DREAMS Partnership, an ambitious package of interventions in 10 sub-Saharan African countries. DREAMS aims to reduce HIV incidence by 40% among AGYW over two years by addressing multiple causes of AGYW vulnerability. This protocol outlines an impact evaluation of DREAMS in four settings. To achieve an impact evaluation that is credible and timely, we describe a mix of methods that build on longitudinal data available in existing surveillance sites prior to DREAMS roll-out. In three long-running surveillance sites (in rural and urban Kenya and rural South Africa), the evaluation will measure: (1) population-level changes over time in HIV incidence and socio-economic, behavioural and health outcomes among AGYW and young men (before, during, after DREAMS); and (2) causal pathways linking uptake of DREAMS interventions to 'mediators' of change such as empowerment, through to behavioural and health outcomes, using nested cohort studies with samples of ~ 1000-1500 AGYW selected randomly from the general population and followed for two years. In Zimbabwe, where DREAMS includes an offer of pre-exposure HIV prophylaxis (PrEP), cohorts of young women who sell sex will be followed for two years to measure the impact of 'DREAMS+PrEP' on HIV incidence among young women at highest risk of HIV. In all four settings, process evaluation and qualitative studies will monitor the delivery and context of DREAMS implementation. The primary evaluation outcome is HIV incidence, and secondary outcomes include indicators of sexual behavior change, and social and biological protection. DREAMS is, to date, the most ambitious effort to scale-up combinations or 'packages' of multi-sectoral interventions for HIV prevention. Evidence of its effectiveness in reducing HIV incidence among AGYW, and demonstrating which aspects of the lives of AGYW were changed, will offer valuable lessons for replication.


Ferroelectric Bi3.25La0.75Ti3O12 films of uniform a-axis orientation on silicon substrates.
The use of bismuth-layered perovskite films for planar-type nonvolatile ferroelectric random-access memories requires films with spontaneous polarization normal to the plane of growth. Epitaxially twinned a axis-oriented La-substituted Bi4Ti3O12 (BLT) thin films whose spontaneous polarization is entirely along the film normal were grown by pulsed laser deposition on yttria-stabilized zirconia-buffered Si(100) substrates using SrRuO3 as bottom electrodes. Even though the (118) orientation competes with the (100) orientation, epitaxial films with almost pure (100) orientation were grown using very thin, strained SrRuO3 electrode layers and kinetic growth conditions, including high growth rates and high oxygen background pressures to facilitate oxygen incorporation into the growing film. Films with the a-axis orientation and having their polarization entirely along the direction normal to the film plane can achieve a remanent polarization of 32 microcoulombs per square centimeter.


Receptor interacting protein kinase-3 determines cellular necrotic response to TNF-alpha.
Smac mimetics induce apoptosis synergistically with TNF-alpha by triggering the formation of a caspase-8-activating complex containing receptor interacting protein kinase-1 (RIPK1). Caspase inhibitors block this form of apoptosis in many types of cells. However, in several other cell lines, caspase inhibitors switch the apoptotic response to necrosis. A genome wide siRNA screen revealed another member of the RIP kinase family, RIP3, to be required for necrosis. The expression of RIP3 in different cell lines correlates with their responsiveness to necrosis induction. The kinase activity of RIP3 is essential for necrosis execution. Upon induction of necrosis, RIP3 is recruited to RIPK1 to form a necrosis-inducing complex. Embryonic fibroblasts from RIP3 knockout mice are resistant to necrosis and RIP3 knockout animals are devoid of inflammation inflicted tissue damage in an acute pancreatitis model. These data indicate RIP3 as the determinant for cellular necrosis in response to TNF-alpha family of death-inducing cytokines.


Reduction or discontinuation of antipsychotics for challenging behaviour in adults with intellectual disability: a systematic review.
The use of antipsychotics to manage challenging behaviour in adults with intellectual disability is widespread but controversial, and evidence is scarce. There is a perception that antipsychotics used in this context can be reduced or discontinued, and this has been a major focus of recent national policy. However, such an intervention risks harm as well as having potential benefits. We reviewed the available evidence and found that antipsychotics can be reduced or discontinued in a substantial proportion of adults who use them for challenging behaviour, although not always without adverse effects. There is a group which displays behavioural deterioration on antipsychotic reduction that prevents discontinuation; predictors of poor response could not be reliably identified. In view of the relatively scarce data and methodological limitations of the available studies, we cannot draw firm conclusions to inform a population level approach to this issue. Antipsychotic medication used for behaviour should be reviewed regularly and an individualised approach taken to treatment.


Assessment of perfusion in the diabetic foot by regional transcutaneous oximetry.
Regional transcutaneous oximetry is a new, noninvasive diagnostic technique for the investigation of peripheral vascular disease (PVD) that uses differences in limb and trunk transcutaneous PO2 to assess the adequacy of local perfusion. The application of such measurements would be of great importance in diabetes, in which limb ischemia is commonly difficult to assess. A group of diabetic subjects with symptomatic PVD was studied with regional oximetry, Doppler-assisted blood pressure measurements, and arteriography. Doppler studies correlated poorly to symptom grade and angiographic data, while oximetry clearly demonstrated limb hypoxia under the functional conditions appropriate to the patients' clinical symptomatology. The superiority of oximetry to Doppler studies was highly significant (X2 = 12.64, P less than 0.001). Regional transcutaneous oximetry should therefore be the noninvasive diagnostic test of choice in the initial evaluation of the diabetic limb for PVD. Because of its dependence on the adequacy of local oxygenation, transcutaneous oximetry is a powerful tool for investigation of the pathophysiology of PVD and will, in the future, have wide-ranging applications to the diagnosis and therapy of PVD.


Glycaemic, blood pressure and lipid goal attainment and chronic kidney disease stage of type 2 diabetic patients treated in primary care practices.
The prevalence of chronic kidney disease and diabetes is rising in Europe. These patients are at high cardiovascular and renal risk and need a challenging multifactorial therapeutic approach. The goal of this cross-sectional study was to examine the treatment and attainments of goals related to cardiovascular risk factors within chronic kidney disease stages in type 2 diabetic patients followed up by primary care physicians in Switzerland. Each participating physician entered into a web database the anonymised data of up to 15 consecutive diabetic patients attending her/his office between December 2013 and June 2014. Diabetes, hypertension and lipid lowering therapies were analysed, as well as glycated haemoglobin (HbA1c), blood pressure and low-density lipoprotein-cholesterol (LDL-c) levels and goal attainments by KDIGO chronic kidney disease stage 1 to 4. A total of 1359 patients (mean age 66.5±12.4 years) were included by 109 primary care physicians. Chronic kidney disease stages 0-2, 3a, 3 b and 4 were present in 77.6%, 13.9%, 6.1%, and 2.4%, respectively. Average HbA1c was independent of chronic kidney disease stage and close to 7%; more than half of the patients reached the HbA1c goal. Eighty-four percent of patients were hypertensive and only 18.2% reached the then current Swiss or American Diabetes Association 2013 blood pressure goals. Despite loosening of blood pressure goals in 2015, only half of the patients reached them and most needed multiple therapies. Increased body mass index and advanced chronic kidney disease stage decreased the chance of reaching blood pressure goals. Lipid lowering therapy was prescribed in 62.1% of cases, with average LDL-c levels similar across chronic kidney disease stages. Only 42% of patients reached the LDL-c goal of <2.5 mmol/l in primary prevention and 32% reached <1.8 mmol/l in secondary prevention. Younger patients were treated significantly less aggressively than older patients (≥68 years, median age) for HbA1c, LDL-c and diastolic blood pressure control. This cross-sectional study demonstrates that blood pressure and lipid goals are less often achieved than blood glucose control in type 2 diabetic patients followed up by primary care physicians in Switzerland. Goal attainments for HbA1c and LDL-c were not influenced by chronic kidney disease stages, in contrast to blood pressure. Reaching all three goals was rare (2.2%). There is a need for improvement in blood pressure control in advanced chronic kidney disease, whereas HbA1c goals may be loosened in the elderly and in advanced chronic kidney disease.


A systematic review of empirical and simulation studies evaluating the health impact of transportation interventions.
Urban transportation is an important determinant of health and environmental outcomes, and therefore essential to achieving the United Nation's Sustainable Development Goals. To better understand the health impacts of transportation initiatives, we conducted a systematic review of longitudinal health evaluations involving: a) bus rapid transit (BRT); b) bicycle lanes; c) Open Streets programs; and d) aerial trams/cable cars. We also synthesized systems-based simulation studies of the health-related consequences of walking, bicycling, aerial tram, bus and BRT use. Two reviewers screened 3302 unique titles and abstracts identified through a systematic search of MEDLINE (Ovid), Scopus, TRID and LILACS databases. We included 39 studies: 29 longitudinal evaluations and 10 simulation studies. Five studies focused on low- and middle-income contexts. Of the 29 evaluation studies, 19 focused on single component bicycle lane interventions; the rest evaluated multi-component interventions involving: bicycle lanes (n = 5), aerial trams (n = 1), and combined bicycle lane/BRT systems (n = 4). Bicycle lanes and BRT systems appeared effective at increasing bicycle and BRT mode share, active transport duration, and number of trips using these modes. Of the 10 simulation studies, there were 9 agent-based models and one system dynamics model. Five studies focused on bus/BRT expansions and incentives, three on interventions for active travel, and the rest investigated combinations of public transport and active travel policies. Synergistic effects were observed when multiple policies were implemented, with several studies showing that sizable interventions are required to significantly shift travel mode choices. Our review indicates that bicycle lanes and BRT systems represent promising initiatives for promoting population health. There is also evidence to suggest that synergistic effects might be achieved through the combined implementation of multiple transportation policies. However, more rigorous evaluation and simulation studies focusing on low- and middle-income countries, aerial trams and Open Streets programs, and a more diverse set of health and health equity outcomes is required.


Do mothers prefer helpers or smaller litters? Birth sex ratio and litter size adjustment in cotton-top tamarins (Saguinus oedipus).
Sex allocation theory has been a remarkably productive field in behavioral ecology with empirical evidence regularly supporting quantitative theoretical predictions. Across mammals in general and primates in particular, however, support for the various hypotheses has been more equivocal. Population-level sex ratio biases have often been interpreted as supportive, but evidence for small-scale facultative adjustment has rarely been found. The helper repayment (HR) also named the local resource enhancement (LRE) hypothesis predicts that, in cooperatively breeding species, mothers invest more in the sex which assists with rearing future offspring and that this bias will be more pronounced in mothers who require extra assistance (i.e., due to inexperience or a lack of available alloparents). We tested these hypotheses in captive cotton-top tamarins (Saguinus oedipus) utilizing the international studbook and birth records obtained through a questionnaire from ISIS-registered institutions. Infant sex, litter size, mother's age, parity, and group composition (presence of nonreproductive subordinate males and females) were determined from these records. The HR hypothesis was supported over the entire population, which was significantly biased toward males (the "helpful" sex). We found little support for helper repayment at the individual level, as primiparous females and those in groups without alloparents did not exhibit more extreme tendencies to produce male infants. Primiparous females were, however, more likely to produce singleton litters. Singleton births were more likely to be male, which suggests that there may be an interaction between litter size adjustment and sex allocation. This may be interpreted as supportive of the HR hypothesis, but alternative explanations at both the proximate and ultimate levels are possible. These possibilities warrant further consideration when attempting to understand the ambiguous results of primate sex ratio studies so far.


Retransplantation of the liver--a seven-year experience.
Three hundred and four patients underwent 362 liver transplants between July 1984 and April 1992. Fifty-eight retransplants were performed in 44 patients (14.5%). Thirty-four patients underwent two (77.3%), seven patients three (15.9%), two patients four (4.5%), and one patient five (2.3%) transplants. Poor function accounted for 23 retransplants (6.4%), technical problems for 19 retransplants (5.2%), and rejection for 15 retransplants (4.1%). One-month patient survivals after retransplantation for poor function, technical problems, or rejection were similar (79.0%, 73.4%, and 80.0%, respectively). No difference in retransplantation rates were seen between adults and children receiving whole liver transplants (WLT) (11.6% versus 19.1%). However, retransplantation for poor function was more common in pediatric recipients receiving reduced-size liver transplants (RLT) (20.0% versus 0.0%, P < 0.01), while retransplantation for hepatic artery thrombosis (HAT) was more common in pediatric recipients receiving WLT (16.7% versus 2.8%, P < 0.05). The presence of multiorgan system failure of greater than four was associated with a high mortality (90%), whereas patients undergoing emergent retransplantation who had less than four systems fail had a survival of 73.9% and patients who underwent elective retransplantation had a survival rate of 81.8%. Length of stay and cost of liver transplantation was higher in patients undergoing retransplantation when compared with primary transplants (29.7 +/- 14.9 days versus 58.4 +/- 38.9 days and $122,358 +/- 59,782 versus $289,302 +/- 126,907, P < 0.01). The overall actuarial one-year patient survival in primary transplants was 86.6% and in retransplants 74.8%, and at five years these were 71.4% versus 62.5%, respectively (P < 0.05). Our results support continued retransplantation of the liver unless the patient's medical condition dictates otherwise.


Rediscovering market segmentation.
In 1964, Daniel Yankelovich introduced in the pages of HBR the concept of nondemographic segmentation, by which he meant the classification of consumers according to criteria other than age, residence, income, and such. The predictive power of marketing studies based on demographics was no longer strong enough to serve as a basis for marketing strategy, he argued. Buying patterns had become far better guides to consumers' future purchases. In addition, properly constructed nondemographic segmentations could help companies determine which products to develop, which distribution channels to sell them in, how much to charge for them, and how to advertise them. But more than 40 years later, nondemographic segmentation has become just as unenlightening as demographic segmentation had been. Today, the technique is used almost exclusively to fulfill the needs of advertising, which it serves mainly by populating commercials with characters that viewers can identify with. It is true that psychographic types like "High-Tech Harry" and "Joe Six-Pack" may capture some truth about real people's lifestyles, attitudes, self-image, and aspirations. But they are no better than demographics at predicting purchase behavior. Thus they give corporate decision makers very little idea of how to keep customers or capture new ones. Now, Daniel Yankelovich returns to these pages, with consultant David Meer, to argue the case for a broad view of nondemographic segmentation. They describe the elements of a smart segmentation strategy, explaining how segmentations meant to strengthen brand identity differ from those capable of telling a company which markets it should enter and what goods to make. And they introduce their "gravity of decision spectrum", a tool that focuses on the form of consumer behavior that should be of the greatest interest to marketers--the importance that consumers place on a product or product category.


Antitrypanosomal 8-Hydroxy-Naphthyridines Are Chelators of Divalent Transition Metals.
The lack of information regarding the mechanisms of action (MoA) or specific molecular targets of phenotypically active compounds can prove a barrier to their development as chemotherapeutic agents. Here, we report the results of our orthogonal genetic, molecular, and biochemical studies to determine the MoA of a novel 7-substituted 8-hydroxy-1,6-naphthyridine (8-HNT) series that displays promising activity against Trypanosoma brucei and Leishmania donovani High-throughput loss-of-function genetic screens in T. brucei highlighted two probable zinc transporters associated with resistance to these compounds. These transporters localized to the parasite Golgi apparatus. Directed by these findings, the role of zinc and other divalent cations in the MoA of these compounds was investigated. 8-HNT compounds were found to directly deplete intracellular levels of Zn2+, while the addition of exogenous Zn2+ and Fe2+ reduced the potency of compounds from this series. Detailed biochemical analyses confirmed that 8-HNT compounds bind directly to a number of divalent cations, predominantly Zn2+, Fe2+, and Cu2+, forming 2:1 complexes with one of these cations. Collectively, our studies demonstrate transition metal depletion, due to chelation, as the MoA of the 8-HNT series of compounds. Strategies to improve the selectivity of 8-HNT compounds are discussed.


Examining the ribonuclease H primer grip of HIV-1 reverse transcriptase by charge neutralization of RNA/DNA hybrids.
The crystal structure of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) bound to an RNA/DNA hybrid reveals an extensive network of contacts with the phosphate backbone of the DNA strand approximately 4-9 bp downstream from the ribonuclease H (RNase H) catalytic center. Collectively designated as 'the RNase H primer grip', this motif contains a phosphate binding pocket analogous to the human and Bacillus halodurans RNases H. The notion that the RNase H primer grip mediates the trajectory of RNA/DNA hybrids accessing the RNase H active site suggests that locally neutralizing the phosphate backbone may be exploited to manipulate nucleic acid flexibility. To examine this, we introduced single and tandem methylphosphonate substitutions through the region of the DNA primer contacted by the RNase H primer grip and into the RNase H catalytic center. The ability of mutant hybrids to support RNase H and DNA polymerase activity was thereafter examined. In addition, site-specific chemical footprinting was used to evaluate movement of the DNA polymerase and RNase H domains. We show here that minor alteration to the RNase H primer can have a dramatic effect on enzyme positioning, and discuss these findings in light of recent crystallography of human RNase H containing an RNA/DNA hybrid.


Cigarette smoke induces DNA damage and alters base-excision repair and tau levels in the brain of neonatal mice.
The prenatal and perinatal periods of brain development are especially vulnerable to insults by environmental agents. Early life exposure to cigarette smoke (CS), which contains both genotoxicants and oxidants, is considered an important risk factor for both neurodevelopmental and neurodegenerative disorders. Yet, little is known regarding the underlying pathogenetic mechanisms. In the present study, neonatal Swiss ICR (CD-1) albino mice were exposed to various concentrations of CS for 4 weeks and the brain examined for lipid peroxides, DNA damage, base-excision repair (BER) enzymes, apoptosis, and levels of the microtubule protein tau. CS induced a dose-dependent increase in both malondialdehyde and various types of DNA damage, including single-strand breaks, double-strand breaks, and DNA-protein cross-links. However, the CS-induced DNA damage in the brain returned to basal levels 1 week after smoking cessation. CS also modulated the activity and distribution of the BER enzymes 8-oxoguanine-DNA-glycosylase (OGG1) and apyrimidinic/apurinic endonuclease (APE1) in several brain regions. Normal tau (i.e., three-repeat tau, 3R tau) and various pathological forms of tau were also measured in the brain of CS-exposed neonatal mice, but only 3R tau and tau phosphorylated at serine 199 were significantly elevated. The oxidative stress, genomic dysregulation, and alterations in tau metabolism caused by CS during a critical period of brain development could explain why CS is an important risk factor for both neurodevelopmental and neurodegenerative disorders appearing in later life.


Targeted expression of cyclin D2 ameliorates late stage anthracycline cardiotoxicity.
Doxorubicin (DOX) is a widely used and effective anti-cancer therapeutic. DOX treatment is associated with both acute and late onset cardiotoxicity, limiting its overall efficacy. Here, the impact of cardiomyocyte cell cycle activation was examined in a juvenile model featuring aspects of acute and late onset DOX cardiotoxicity. Two-week old MHC-cycD2 transgenic mice (which express cyclin D2 in postnatal cardiomyocytes and exhibit sustained cardiomyocyte cell cycle activity; D2 mice) and their wild type (WT) littermates received weekly DOX injections for 5 weeks (25 mg/kg cumulative dose). One week after the last DOX treatment (acute stage), cardiac function was suppressed in both groups. Acute DOX cardiotoxicity in D2 and WT mice was associated with similar increases in the levels of cardiomyocyte apoptosis and Ku70/Ku80 expression (markers of DNA damage and oxidative stress), as well as similar reductions in hypertrophic cardiomyocyte growth. Cardiac dysfunction persisted in WT mice for 13 weeks following the last DOX treatment (late stage) and was accompanied by increased levels of cardiomyocyte apoptosis, Ku expression, and myocardial fibrosis. In contrast, D2 mice exhibited a progressive recovery in cardiac function, which was indistinguishable from saline-treated animals by 9 weeks following the last DOX treatment. Improved cardiac function was accompanied by reductions in the levels of late stage cardiomyocyte apoptosis, Ku expression, and myocardial fibrosis. These data suggest that cardiomyocyte cell cycle activity can promote recovery of cardiac function and preserve cardiac structure following DOX treatment.


Persistence of self-recruitment and patterns of larval connectivity in a marine protected area network.
The use of marine protected area (MPA) networks to sustain fisheries and conserve biodiversity is predicated on two critical yet rarely tested assumptions. Individual MPAs must produce sufficient larvae that settle within that reserve's boundaries to maintain local populations while simultaneously supplying larvae to other MPA nodes in the network that might otherwise suffer local extinction. Here, we use genetic parentage analysis to demonstrate that patterns of self-recruitment of two reef fishes (Amphiprion percula and Chaetodon vagabundus) in an MPA in Kimbe Bay, Papua New Guinea, were remarkably consistent over several years. However, dispersal from this reserve to two other nodes in an MPA network varied between species and through time. The stability of our estimates of self-recruitment suggests that even small MPAs may be self-sustaining. However, our results caution against applying optimization strategies to MPA network design without accounting for variable connectivity among species and over time.


Non-IG aberrations of FOXP1 in B-cell malignancies lead to an aberrant expression of N-truncated isoforms of FOXP1.
The transcription factor FOXP1 is implicated in the pathogenesis of B-cell lymphomas through chromosomal translocations involving either immunoglobulin heavy chain (IGH) locus or non-IG sequences. The former translocation, t(3;14)(p13;q32), results in dysregulated expression of FOXP1 juxtaposed with strong regulatory elements of IGH. Thus far, molecular consequences of rare non-IG aberrations of FOXP1 remain undetermined. Here, using molecular cytogenetics and molecular biology studies, we comprehensively analyzed four lymphoma cases with non-IG rearrangements of FOXP1 and compared these with cases harboring t(3;14)(p13;q32)/IGH-FOXP1 and FOXP1-expressing lymphomas with no apparent structural aberrations of the gene. Our study revealed that non-IG rearrangements of FOXP1 are usually acquired during clinical course of various lymphoma subtypes, including diffuse large B cell lymphoma, marginal zone lymphoma and chronic lymphocytic leukemia, and correlate with a poor prognosis. Importantly, these aberrations constantly target the coding region of FOXP1, promiscuously fusing with coding and non-coding gene sequences at various reciprocal breakpoints (2q36, 10q24 and 3q11). The non-IG rearrangements of FOXP1, however, do not generate functional chimeric genes but commonly disrupt the full-length FOXP1 transcript leading to an aberrant expression of N-truncated FOXP1 isoforms (FOXP1(NT)), as shown by QRT-PCR and Western blot analysis. In contrast, t(3;14)(p13;q32)/IGH-FOXP1 affects the 5' untranslated region of FOXP1 and results in overexpress the full-length FOXP1 protein (FOXP1(FL)). RNA-sequencing of a few lymphoma cases expressing FOXP1(NT) and FOXP1(FL) detected neither FOXP1-related fusions nor FOXP1 mutations. Further bioinformatic analysis of RNA-sequencing data retrieved a set of genes, which may comprise direct or non-direct targets of FOXP1(NT), potentially implicated in disease progression. In summary, our findings point to a dual mechanism through which FOXP1 is implicated in B-cell lymphomagenesis. We hypothesize that the primary t(3;14)(p13;q32)/IGH-FOXP1 activates expression of the FOXP1(FL) protein with potent oncogenic activity, whereas the secondary non-IG rearrangements of FOXP1 promote expression of the FOXP1(NT) proteins, likely driving progression of disease.


The murine MHC class I genes, H-2Dq and H-2Lq, are strikingly homologous to each other, H-2Ld, and two genes reported to encode tumor-specific antigens.
Two phenomena appear to distinguish the D region class I genes from those in the K region in the murine MHC: (a) haplotype disparity in the number of expressed D region class I molecules has been observed; and (b) clines of closely related D region class I molecules among and within mice of different H-2 haplotypes can be defined. Both of these observations have been based on serological and peptide mapping analyses of these molecules. Recent reports using molecular biological approaches have corroborated these findings. Since the mouse strain B10.AKM expresses multiple D region class I antigens, all of which are closely related to the prototypic Ld molecule, we investigated the Dq region of B10.AKM using molecular approaches. Three D region class I genes were isolated from genomic B10.AKM bacteriophage and cosmid libraries. Based on alignment of those genes with the BALB/c D region class I genes by analogous restriction endonuclease sites and by hybridization of one of those genes with a D4d gene-derived oligonucleotide probe, we have designated these genes as Dq, Lq, and D4q. As determined by DNA-mediated gene transfer to mouse L cells followed by serological analyses, the Dq and Lq genes encode previously characterized Dq region class I antigens. The nucleic acid sequence comparisons of the Dq and Lq genes demonstrated a higher level of homology with the Ld and Db genes than with other D region class I genes. In addition, CTL stimulated with a Dq, Lq, or Ld gene transfectant showed strong crossreactions with the other transfectants as targets, suggesting that the products of these genes are also functionally related. Thus, these studies suggest that the L molecule represents a prototypic structure shared by several D region gene products, and furthermore, the duplication of an Ld-like progenitor gene resulted in two Dq region class I genes, Dq and Lq. Unexpectedly, the sequences determined for the Dq and Lq genes are nearly identical to the sequences of two genes, A166 and A149, respectively, which were reported to encode the tumor-specific antigens; these novel class I genes were isolated from an H-2k fibrosarcoma, 1591. This raises the distinct possibility that these purported tumor-specific class I genes were introduced into this tumor by contamination.


Deletion of estrogen receptors reveals a regulatory role for estrogen receptors-beta in bone remodeling in females but not in males.
To determine the contributions of estrogen receptor (ER)alpha and ERbeta in bone growth and remodeling in male and female mice, we generated and analyzed full knockouts for each receptor, and a double ER knockout. Although suppression of the ligand to the ERs (i.e., estradiol) after menopause or gonadectomy in females led to a catastrophic increase in bone turnover and concomitant bone loss, deletion of one or both ERs failed to show such an effect. Complete deletion of ERalpha led to a decrease, not an increase, in bone turnover and an increase, not a decrease, in trabecular bone volume in both male and female animals. Deletion of ERbeta led to different responses in males, where bone was unaffected, and in females, where bone resorption was decreased and trabecular bone volume increased. In contrast, deletion of both ERs led to a profound decrease in trabecular bone volume in females, which was associated with a decrease, not an increase, in bone turnover. Finally, deletion of ERalpha, but not ERbeta, led to major changes in circulating levels of estradiol and/or testosterone, indirectly affecting bone remodeling and bone mass. Thus, only ERalpha was shown to regulate bone remodeling in males, whereas in females both receptor subtypes influenced this process and could, at least under basal knockout conditions, compensate for each other.


The relationship between the molecular structure of natural acetogenins and their inhibitory activities which affect DNA polymerase, DNA topoisomerase and human cancer cell growth.
Acetogenins from the Annonaceous plant are a fatty acid-derived natural product. Chemically synthesized natural acetogenins, such as mucocin (compound 1), jimenezin (compound 2), muconin (compound 4), pyranicin (compound 5) and pyragonicin (compound 6) were investigated. Concomitantly, 19-epi jimenezin (compound 3), 10-epi pyragonicin (compound 7) and a γ-lactone (compound 8), which is estimated to be a biosynthetic precursor of acetogenins, were synthesized and investigated. Compounds 5 and 6 strongly inhibited, and compound 7 moderately inhibited the activities of mammalian DNA polymerases (pols), such as replicative pol α and repair/recombination-related pol β and λ, and also inhibited human DNA topoisomerase (topos) I and II activities. On the other hand, compounds 1-4 and 8 did not influence the activities of any pols and topos. Compound 5 was the strongest inhibitor of the pols and topos tested, and the IC(50) values were 5.0-9.6 μM, respectively. These compounds also suppressed human cancer cell growth with almost the same tendency as the inhibition of pols and topos. Compound 5 was the strongest suppressor of the proliferation of the promyelocytic leukemia cell line, HL-60, in human cancer cell lines tested with an LD(50) value of 9.4 μM, and arrested the cells at G1 phases, indicating that it blocks DNA replication by inhibiting the activity of pols rather than topos. This compound also induced cell apoptosis. The relationship between the three-dimensional molecular structure of acetogenins and these inhibitory activities is discussed. The results suggested that compound 5 is a lead compound of potentially useful cancer chemotherapy agents.


Loss of ovarian function in the VCD mouse-model of menopause leads to insulin resistance and a rapid progression into the metabolic syndrome.
Factors comprising the metabolic syndrome occur with increased incidence in postmenopausal women. To investigate the effects of ovarian failure on the progression of the metabolic syndrome, female B(6)C(3)F(1) mice were treated with 4-vinylcyclohexene diepoxide (VCD) and fed a high-fat (HF) diet for 16 wk. VCD destroys preantral follicles, causing early ovarian failure and is a well-characterized model for the gradual onset of menopause. After 12 wk on a HF diet, VCD-treated mice had developed an impaired glucose tolerance, whereas cycling controls were unaffected [12 wk AUC HF mice 13,455 +/- 643 vs. HF/VCD 17,378 +/- 1140 mg/dl/min, P < 0.05]. After 16 wk on a HF diet, VCD-treated mice had significantly higher fasting insulin levels (HF 5.4 +/- 1.3 vs. HF/VCD 10.1 +/- 1.4 ng/ml, P < 0.05) and were significantly more insulin resistant (HOMA-IR) than cycling controls on a HF diet (HF 56.2 +/- 16.7 vs. HF/VCD 113.1 +/- 19.6 mg/dl x microU/ml, P < 0.05). All mice on a HF diet gained more weight than mice on a standard diet, and weight gain in HF/VCD mice was significantly increased compared with HF cycling controls. Interestingly, even without a HF diet, progression into VCD-induced menopause caused a significant increase in cholesterol and free fatty acids. Furthermore, in mice fed a standard diet (6% fat), insulin resistance developed 4 mo after VCD-induced ovarian failure. Insulin resistance following ovarian failure (menopause) was prevented by estrogen replacement. Studies here demonstrate that ovarian failure (menopause) accelerates progression into the metabolic syndrome and that estrogen replacement prevents the onset of insulin resistance in VCD-treated mice. Thus, the VCD model of menopause provides a physiologically relevant means of studying how sex hormones influence the progression of the metabolic syndrome.


Gastrectomy and subsequent risk of oesophageal cancer in Milan.
To analyse quantitatively the relationship between gastrectomy and oesophageal cancer risk. This was a case-control study, based on a structured questionnaire that included a problem-oriented medical history. A network of hospitals in the Greater Milan area between 1984 and 1992. Subjects were 316 incident, histologically confirmed cases of oesophageal cancer (258 males and 58 females), and 1408 controls (1031 males, 377 females), admitted to hospital for acute, non-neoplastic, non-digestive tract conditions. Relative risks (RR) and the corresponding 95% confidence intervals (CI) adjusted for age and sex, plus area of residence, education, tobacco, and alcohol consumption. There was an association of borderline significance of oesophageal cancer with gastric ulcer (RR = 1.6), but not with duodenal ulcer (RR = 0.9). Among cases, 25 (7.9%) reported a history of gastrectomy, versus 29 (2.1%) of the controls. The corresponding multivariate RR was 3.8 (95% CI 2.0, 7.0). The RR of oesophageal cancer was significantly raised (RR = 3.6) within 20 years after gastrectomy, and the association seemed stronger with increasing time since gastrectomy (RR = 4.2). Since gastrectomy may cause gastric hypoacidity and favour oesophageal reflux, and hence oesophagitis, the results of this study, if confirmed, are of interest to a better understanding of the mechanisms of oesophageal carcinogenesis.


Visualization of interaction between inorganic nanoparticles and bacteria or fungi.
The objective of the present investigation was to evaluate the morphologic characteristics of self-assemblies of diamond (nano-D), silver (nano-Ag), gold (nano-Au), and platinum (nano-Pt) nanoparticles with Staphylococcus aureus (bacteria) and Candida albicans (fungi), to determine the possibility of constructing microorganism-nanoparticle vehicles. Hydrocolloids of individual nanoparticles were added to suspensions of S. aureus and C. albicans. Immediately after mixing, the samples were inspected by transmission electron microscopy. Visualization of the morphologic interaction between the nanoparticles and microorganisms showed that nano-D, which are dielectrics and exhibit a positive zeta potential, were very different from the membrane potentials of microorganisms, and uniformly surrounded the microorganisms, without causing visible damage and destruction of cells. All metal nanoparticles with negative zeta potential had cell damaging properties. Nano-Ag showed the properties of self-organization with the cells, disintegrating the cell walls and cytoplasmic membranes, and releasing a substance (probably cytoplasm) outside the cell. Arrangement of nano-Au with microorganisms did not create a system of self-organization, but instead a "noncontact" interaction between the nanoparticles and microorganisms was observed to cause damage to fungal cells. Nano-Pt caused both microorganisms to release a substance outside the cell and disintegrated the cytoplasmic membrane and cell wall. Nano-Ag, nano-Au, and nano-Pt (all metal nanoparticles) are harmful to bacteria and fungi. In contrast, nano-D bind closely to the surface of microorganisms without causing visible damage to cells, and demonstrating good self-assembling ability. The results indicate that both microorganisms could be used as potential carriers for nano-D.


Is executive cognitive function associated with youth gambling?
Our objectives for this report were to identify trajectories of youth gambling behavior, and to examine their relation to executive cognitive function (ECF) and associated problem behaviors. Philadelphia school children, enrolled at ages 10-12 years (n = 387; 49% male), completed three annual assessments of risk behaviors, ECF, impulsivity, problem behaviors and demographics. Across ages 10-15 years, using methods from Nagin et al., two groups were identified: Early Gamblers (n = 111) initiated early and continued in later assessments, and Later Gamblers (n = 276) initiated at later ages and gambled less. Betting money on cards and sports were the most frequently reported gambling behaviors. Using gambling group as outcome, final backward selection logistic regression model showed Early Gamblers are more likely male (P = 0.001), report more active coping (P = 0.042), impulsive behaviors (P ≤ 0.008), and have friends who gamble (P = 0.001). Groups were similar in ECF, parental monitoring, marital status, SES, and race. Early Gamblers had higher incidence of problem behaviors and drug use (all P ≤ 0.006). Two gambling groups were identified in early adolescence with Early Gamblers showing higher levels of impulsivity and comorbid problems but similar levels of ECF compared to Late Gamblers. As more gambling groups are identified through later adolescence, ECF may emerge as a relevant precursor of problem gambling at this later time.


The value of coordinated management of interacting ecosystem services.
Coordinating decisions and actions among interacting sectors is a critical component of ecosystem-based management, but uncertainty about coordinated management's effects is compromising its perceived value and use. We constructed an analytical framework for explicitly calculating how coordination affects management decisions, ecosystem state and the provision of ecosystem services in relation to ecosystem dynamics and socio-economic objectives. The central insight is that the appropriate comparison strategy to optimal coordinated management is optimal uncoordinated management, which can be identified at the game theoretic Nash equilibrium. Using this insight we can calculate coordination's effects in relation to uncoordinated management and other reference scenarios. To illustrate how this framework can help identify ecosystem and socio-economic conditions under which coordination is most influential and valuable, we applied it to a heuristic case study and a simulation model for the California Current Marine Ecosystem. Results indicate that coordinated management can more than double an ecosystem's societal value, especially when sectors can effectively manipulate resources that interact strongly. However, societal gains from coordination will need to be reconciled with observations that it also leads to strategic simplification of the ecological food web, and generates both positive and negative impacts on individual sectors and non-target species.


Effects of Huanglian-Jie-Du-Tang and its modified formula on the modulation of amyloid-β precursor protein processing in Alzheimer's disease models.
Huanglian-Jie-Du-Tang (HLJDT) is a famous traditional Chinese herbal formula that has been widely used clinically to treat cerebral ischemia. Recently, we found that berberine, a major alkaloid compound in HLJDT, reduced amyloid-β (Aβ) accumulation in an Alzheimer's disease (AD) mouse model. In this study, we compared the effects of HLJDT, four single component herbs of HLJDT (Rhizoma coptidis (RC), Radix scutellariae (RS), Cortex phellodendri (CP) and Fructus gardenia (FG)) and the modified formula of HLJDT (HLJDT-M, which is free of RS) on the regulatory processing of amyloid-β precursor protein (APP) in an in vitro model of AD. Here we show that treatment with HLJDT-M and its components RC, CP, and the main compound berberine on N2a mouse neuroblastoma cells stably expressing human APP with the Swedish mutation (N2a-SwedAPP) significantly decreased the levels of full-length APP, phosphorylated APP at threonine 668, C-terminal fragments of APP, soluble APP (sAPP)-α and sAPPβ-Swedish and reduced the generation of Aβ peptide in the cell lysates of N2a-SwedAPP. HLJDT-M showed more significant APP- and Aβ- reducing effects than berberine, RC or CP treatment alone. In contrast, HLJDT, its component RS and the main active compound of RS, baicalein, strongly increased the levels of all the metabolic products of APP in the cell lysates. The extract from FG, however, did not influence APP modulation. Interestingly, regular treatment of TgCRND8 APP transgenic mice with baicalein exacerbated the amyloid plaque burden, APP metabolism and Aβ production. Taken together, these data provide convincing evidence that HLJDT and baicalein treatment can increase the amyloidogenic metabolism of APP which is at least partly responsible for the baicalein-mediated Aβ plaque increase in the brains of TgCRND8 mice. On the other hand, HLJDT-M significantly decreased all the APP metabolic products including Aβ. Further study of HLJDT-M for therapeutic use in treating AD is warranted.


Matching 2.5D face scans to 3D models.
The performance of face recognition systems that use two-dimensional images depends on factors such as lighting and subject's pose. We are developing a face recognition system that utilizes three-dimensional shape information to make the system more robust to arbitrary pose and lighting. For each subject, a 3D face model is constructed by integrating several 2.5D face scans which are captured from different views. 2.5D is a simplified 3D (x, y, z) surface representation that contains at most one depth value (z direction) for every point in the (x, y) plane. Two different modalities provided by the facial scan, namely, shape and texture, are utilized and integrated for face matching. The recognition engine consists of two components, surface matching and appearance-based matching. The surface matching component is based on a modified Iterative Closest Point (ICP) algorithm. The candidate list from the gallery used for appearance matching is dynamically generated based on the output of the surface matching component, which reduces the complexity of the appearance-based matching stage. Three-dimensional models in the gallery are used to synthesize new appearance samples with pose and illumination variations and the synthesized face images are used in discriminant subspace analysis. The weighted sum rule is applied to combine the scores given by the two matching components. Experimental results are given for matching a database of 200 3D face models with 598 2.5D independent test scans acquired under different pose and some lighting and expression changes. These results show the feasibility of the proposed matching scheme.


Pilot study on activities of daily living limitations in adults with hemianopsia.
I sought to obtain a general understanding of the limitations in activities of daily living (ADLs) experienced by adults with hemianopsia and quandrantanopsia from acquired brain injury. A semistructured interview addressing the perception of difficulty completing ADLs because of vision loss and a reading performance test were administered to 46 participants referred to a low vision rehabilitation program. Participants reported difficulty completing two basic ADLs, personal hygiene-grooming and feeding, and several instrumental ADLs, including driving, shopping, financial management, telephone usage, and meal preparation. ADL limitations appear to be related to the amount of visual search needed to complete the ADL and disruption of the performance skills of reading, writing, and mobility. People with hemianopsia experience limitations in specific ADLs as a direct result of the disruption of the visual field. Further investigation is needed to confirm these findings and explore interventions to improve ADL performance.


Degradation of mesh coatings and intraperitoneal adhesion formation in an experimental model.
In laparoscopic ventral hernia repair a mesh is placed in direct contact with the viscera, often leading to substantial adhesions. In this experimental study the ability of different coated and uncoated meshes to attenuate adhesion formation was examined. Six commercially available meshes were placed intraperitoneally against a closed peritoneum in rats: Prolene (polypropylene), Timesh and Ultrapro (polypropylene composites with titanium and polyglecaprone respectively), Proceed and Parietex Composite (polypropylene and polyester meshes coated with a layer of cellulose and collagen respectively) and C-Qur (polypropylene mesh coated with a layer of omega-3 fatty acids). Adhesions and incorporation were evaluated macroscopically and microscopically after 7 and 30 days. Parietex Composite and C-Qur significantly reduced adhesion formation at 7 days' follow-up compared with all other meshes. By 30 days, this effect had diminished as a significant increase in adhesions together with phagocytosis of the coating was seen for all meshes with layered coatings (Proceed, Parietex Composite and C-Qur. Incorporation was insufficient for all meshes. The absorbable layers of Parietex Composite and C-Qur reduce adhesion formation to intraperitoneal mesh in the short term, but the effect diminishes and phagocytosis of absorbable coatings may contribute to adhesion formation.


Impact of Acellular Dermal Matrix on Postsurgical Wound Fluid Biomarkers in Prosthetic Breast Reconstruction.
Despite the widespread practice of using biologic scaffolds for soft tissue reinforcement over prosthetic implants, the impact of acellular dermal matrix (ADM) on surgical wound fluid biomarkers over the initial postoperative period after prosthetic breast reconstruction remains poorly understood. Patients undergoing prosthetic breast reconstruction surgery where ADM was likely to be used were consented to have fluid samples collected from surgical drains after surgery. Sample collections occurred at an "Early" time point at 24 to 48 hours after surgery and then a "Late" time point approximately 1 to 2 weeks after surgery. All procedures were performed by a single surgeon. Acellular dermal matrix was placed when prosthetic coverage with autologous tissue could not be achieved. Laboratory analyses were performed in blinded fashion without the knowledge of whether the samples came from the ADM "Present" or "Not Present" group. Twenty-one patients were in the ADM Present group and 18 patients were in the Not Present group. Both groups showed similar demographics based on age and body mass index. Analyses for cell concentration, protein concentration, extracellular matrix protein levels, cell proliferation activity, and matrix metalloproteinase activity showed no significant differences between wound fluid samples from the 2 groups. The presence of ADM does not appear to significantly impact wound biomarkers in prosthetic breast reconstruction. The current study provides useful data regarding the impact of ADM on surgical wound fluid during the initial postoperative period, laying important groundwork for more extensive future studies on the impact of biologic scaffolds on wound biology.


Wild juvenile salmonids in Muchalat Inlet, British Columbia, Canada: factors associated with sea lice prevalence.
The Muchalat Inlet, British Columbia, is among the most westerly points at which aquaculture is practiced in Canada. In this paper, we summarise data from over 18000 wild fish sampled at 16 sites over an 8 yr period, between 2004 and 2011. The most prevalent wild species was chum salmon Oncorhynchus keta (82.4%), followed by Chinook O. tshawytscha (10%) and coho O. kisutch (4.3%). However, inter-annual and seasonal variation was evident, and smaller numbers of other Pacific salmon and stickleback species were sporadically observed. A high percentage of wild salmon (~95%) had no sea lice parasites present, with less than 1% of the fish hosting a mobile-stage sea louse. Of the data for which sea lice species were recorded, just over 96% of samples were identified as Lepeophtheirus salmonis. Logistic regression models assessed the association between the presence of lice and a range of independent variables. These models indicated a significant degree of spatial variation, much of which could be explained in terms of salinity levels. There were also important variations through time, both over the season within a year and across years. In addition, coho salmon were significantly more likely (odds ratio = 1.65; 95% CI = 1.20-2.3) to be infected than chum salmon. The protective effect of low salinity was most clearly seen at values lower than 15 psu, although this was dependent on fish species.


Ultra-high-resolution 3D imaging of atherosclerosis in mice with synchrotron differential phase contrast: a proof of concept study.
The goal of this study was to investigate the performance of 3D synchrotron differential phase contrast (DPC) imaging for the visualization of both macroscopic and microscopic aspects of atherosclerosis in the mouse vasculature ex vivo. The hearts and aortas of 2 atherosclerotic and 2 wild-type control mice were scanned with DPC imaging with an isotropic resolution of 15 μm. The coronary artery vessel walls were segmented in the DPC datasets to assess their thickness, and histological staining was performed at the level of atherosclerotic plaques. The DPC imaging allowed for the visualization of complex structures such as the coronary arteries and their branches, the thin fibrous cap of atherosclerotic plaques as well as the chordae tendineae. The coronary vessel wall thickness ranged from 37.4 ± 5.6 μm in proximal coronary arteries to 13.6 ± 3.3 μm in distal branches. No consistent differences in coronary vessel wall thickness were detected between the wild-type and atherosclerotic hearts in this proof-of-concept study, although the standard deviation in the atherosclerotic mice was higher in most segments, consistent with the observation of occasional focal vessel wall thickening. Overall, DPC imaging of the cardiovascular system of the mice allowed for a simultaneous detailed 3D morphological assessment of both large structures and microscopic details.


Pathogenic Role of PPARα Downregulation in Corneal Nerve Degeneration and Impaired Corneal Sensitivity in Diabetes.
The purpose of this study was to investigate the protective role of peroxisome proliferator-activated receptor α (PPARα) against diabetic keratopathy and corneal neuropathy. Corneal samples were obtained from human donors with and without diabetes. Streptozotocin-induced diabetic rats and mice were orally treated with PPARα agonist fenofibrate. As shown by immunohistochemistry and Western blotting, PPARα was downregulated in the corneas of humans with diabetes and diabetic rats. Immunostaining of β-III tubulin demonstrated that corneal nerve fiber metrics were decreased significantly in diabetic rats and mice, which were partially prevented by fenofibrate treatment. As evaluated using a Cochet-Bonnet aesthesiometer, corneal sensitivity was significantly decreased in diabetic mice, which was prevented by fenofibrate. PPARα -/- mice displayed progressive decreases in the corneal nerve fiber density. Consistently, corneal sensitivity was decreased in PPARα -/- mice relative to wild-type mice by 21 months of age. Diabetic mice showed increased incidence of spontaneous corneal epithelial lesion, which was prevented by fenofibrate while exacerbated by PPARα knockout. Western blot analysis revealed significantly altered neurotrophic factor levels in diabetic rat corneas, which were partially restored by fenofibrate treatment. These results indicate that PPARα protects the corneal nerve from degeneration induced by diabetes, and PPARα agonists have therapeutic potential in the treatment of diabetic keratopathy.


Gemtuzumab ozogamicin: first clinical experiences in children with relapsed/refractory acute myeloid leukemia treated on compassionate-use basis.
Gemtuzumab ozogamicin (GO; Mylotarg) was developed to treat CD33(+) acute myeloid leukemia (AML). To date, only studies in adults and preliminary data from a phase 1 study in children have been reported. We report data on 15 children with relapsed/refractory CD33(+) AML who were treated with GO monotherapy on compassionate use basis (4-9 mg/m(2) up to 3 courses). Eight children showed a reduction in bone marrow blasts to 5% or less, including 5 in complete remission without full platelet recovery (CRp). Three of the 5 children with CRp received transplants almost directly following the last GO course, without awaiting further platelet regeneration. Hence in these children no clear discrimination between complete remission (CR) and CRp could be made. In 6 of 8 responding patients further treatment was given consisting of stem cell transplantation (SCT). Two patients are still alive, currently 6 and 9 months after SCT. Hematologic toxicity was difficult to assess due to subsequent SCT or leukemia. Side effects, in one patient each included veno-occlusive disease, transient grade 3 hyperbilirubinemia, transient grade 3 transaminase elevation, and grade 3 hypotension during GO administration. No infections or mucositis occurred. This report demonstrates clinical efficacy of GO in a subset of relapsed/refractory pediatric CD33(+) AML patients and suggests that intensive postremission therapy after remission induction by GO may result in durable responses in some patients, although follow-up is still short. Further studies are needed to determine the efficacy and safety of GO in children with AML.


Polarization of skylight in the O(2)A band: effects of aerosol properties.
Motivated by several observations of the degree of linear polarization of skylight in the oxygen A (O(2)A) band that do not yet have a quantitative explanation, we analyze the influence of aerosol altitude, microphysics, and optical thickness on the degree of linear polarization of the zenith skylight in the spectral region of the O(2)A band, between 755 to 775 nm. It is shown that the degree of linear polarization inside the O(2)A band is particularly sensitive to aerosol altitude. The sensitivity is strongest for aerosols within the troposphere and depends also on their microphysical properties and optical thickness. The polarization of the O(2)A band can be larger than the polarization of the continuum, which typically occurs for strongly polarizing aerosols in an elevated layer, or smaller, which typically occurs for depolarizing aerosols or cirrus clouds in an elevated layer. We show that in the case of a single aerosol layer in the atmosphere a determination of the aerosol layer altitude may be obtained. Furthermore, we show limitations of the aerosol layer altitude determination in case of multiple aerosol layers. To perform these simulations we developed a fast method for multiple scattering radiative transfer calculations in gaseous absorption bands including polarization. The method is a combination of doubling-adding and k-binning methods. We present an error estimation of this method by comparing with accurate line-by-line radiative transfer simulations. For the Motivated by several observations of the degree of linear polarization of skylight in the oxygen A (O(2)A) band that do not yet have a quantitative explanation, we analyze the influence of aerosol altitude, microphysics, and optical thickness on the degree of linear polarization of the zenith skylight in the spectral region of the O(2)A band, between 755 to 775 nm. It is shown that the degree of linear polarization inside the O(2)A band is particularly sensitive to aerosol altitude. The sensitivity is strongest for aerosols within the troposphere and depends also on their microphysical properties and optical thickness. The polarization of the O(2)A band can be larger than the polarization of the continuum, which typically occurs for strongly polarizing aerosols in an elevated layer, or smaller, which typically occurs for depolarizing aerosols or cirrus clouds in an elevated layer. We show that in the case of a single aerosol layer in the atmosphere a determination of the aerosol layer altitude may be obtained. Furthermore, we show limitations of the aerosol layer altitude determination in case of multiple aerosol layers. To perform these simulations we developed a fast method for multiple scattering radiative transfer calculations in gaseous absorption bands including polarization. The method is a combination of doubling-adding and k-binning methods. We present an error estimation of this method by comparing with accurate line-by-line radiative transfer simulations. For the O(2)A band, the errors in the degree of linear polarization are less than 0.11% for transmitted light, and less than 0.31% for reflected light. band, the errors in the degree of linear polarization are less than 0.11% for transmitted light, and less than 0.31% for reflected light.