Identification of a new enamovirus associated with citrus vein enation disease by deep sequencing of small RNAs.
To identify the causal agent of citrus vein enation disease, we examined by deep sequencing (Solexa-Illumina) the small RNA (sRNA) fraction from infected and healthy Etrog citron plants. Our results showed that virus-derived sRNAs (vsRNAs): (i) represent about 14.21% of the total sRNA population, (ii) are predominantly of 21 and 24 nucleotides with a biased distribution of their 5' nucleotide and with a clear prevalence of those of (+) polarity, and (iii) derive from all the viral genome, although a prominent hotspot is present at a 5'-proximal region. Contigs assembled from vsRNAs showed similarity with luteovirus sequences, particularly with Pea enation mosaic virus, the type member of the genus Enamovirus. The genomic RNA (gRNA) sequence of a new virus, provisionally named Citrus vein enation virus (CVEV), was completed and characterized. The CVEV gRNA was found to be single-stranded, positive-sense, with a size of 5,983 nucleotides and five open reading frames. Phylogenetic comparisons based on amino acid signatures of the RNA polymerase and the coat protein clearly classifies CVEV within the genus Enamovirus. Dot-blot hybridization and reverse transcription-polymerase chain reaction tests were developed to detect CVEV in plants affected by vein enation disease. CVEV detection by these methods has already been adopted for use in the Spanish citrus quarantine, sanitation, and certification programs.


Desert tortoises (Gopherus agassizii) are selective herbivores that track the flowering phenology of their preferred food plants.
Previous studies of desert tortoise foraging ecology in the western Mojave Desert suggest that these animals are selective herbivores, which alter their diet according to the temporal availability of preferred food plants. These studies, however, did not estimate availability of potential food plants by taking into account the spatial and temporal variability in ephemeral plant abundance that occurs within the spring season. In this study, we observed 18 free-ranging adult tortoises take 35,388 bites during the spring foraging season. We also estimated the relative abundance of potential food plants by stratifying our sampling across different phenological periods of the 3-month long spring season and by different habitats and microhabitats. This methodology allowed us to conduct statistical tests comparing tortoise diet against plant abundance. Our results show that tortoises choose food plants non-randomly throughout the foraging season, a finding that corroborates the hypothesis that desert tortoises rely on key plants during different phenological periods of spring. Moreover, tortoises only consumed plants in a succulent state until the last few weeks of spring, at which time most annuals and herbaceous perennials had dried and most tortoises had ceased foraging. Many species of food plants--including several frequently eaten species--were not detected in our plant surveys, yet tortoises located these rare plants in their home ranges. Over 50% of bites consumed were in the group of undetected species. Interestingly, tortoises focused heavily on several leguminous species, which could be nutritious foods owing to their presumably high nitrogen contents. We suggest that herbaceous perennials, which were rare on our study area but represented ~30% of tortoise diet, may be important in sustaining tortoise populations during droughts when native annuals are absent. These findings highlight the vulnerability of desert tortoises to climate change if such changes alter the availability of their preferred food plants.


The association of depression and anxiety with pain: a study from NESDA.
Chronic pain is commonly co-morbid with a depressive or anxiety disorder. Objective of this study is to examine the influence of depression, along with anxiety, on pain-related disability, pain intensity, and pain location in a large sample of adults with and without a depressive and/or anxiety disorder. The study population consisted of 2981 participants with a depressive, anxiety, co-morbid depressive and anxiety disorder, remitted disorder or no current disorder (controls). Severity of depressive and anxiety symptoms was also assessed. In separate multinomial regression analyses, the association of presence of depressive or anxiety disorders and symptom severity with the Chronic Pain Grade and location of pain was explored. Presence of a depressive (OR = 6.67; P<.001), anxiety (OR = 4.84; P<.001), or co-morbid depressive and anxiety disorder (OR = 30.26; P<.001) was associated with the Chronic Pain Grade. Moreover, symptom severity was associated with more disabling and severely limiting pain. Also, a remitted depressive or anxiety disorder showed more disabling and severely limiting pain (OR = 3.53; P<.001) as compared to controls. A current anxiety disorder (OR = 2.96; p<.001) and a co-morbid depressive and anxiety disorder (OR = 5.15; P<.001) were more strongly associated with cardio-respiratory pain, than gastro-intestinal or musculoskeletal pain. These findings remain after adjustment for chronic cardio respiratory illness. Patients with a current and remitted depressive and/or anxiety disorder and those with more severe symptoms have more disabling pain and pain of cardio-respiratory nature, than persons without a depressive or anxiety disorder. This warrants further research.


The evolving health-care environment: new arguments for closer collaboration between cardiac surgical intensive-care nurses and clinical engineers.
"Not vain the weakest, if their force unite." Homer, Iliad (9th century, B.C.). In today's evolving health-care environment, the cardiac surgical intensive-care nurse is required to provide care to patients whose acuity levels are increasing. Simultaneously, these nurses are asked to use more technologically complex devices to deliver that care. In addition, practice protocols are being integrated into hospitals' clinical information systems. To meet these challenges, cardiac surgical intensive-care nurses must collaborate with clinical engineers on the evaluation and installation of new products and on the automation and refinement of clinical pathways and other outcomes measurement tools. Each discipline also must ensure that the other keeps pace with and maintains its levels of proficiency in the technology used to care for patients undergoing cardiac surgery.


A Snf2 family ATPase complex required for recruitment of the histone H2A variant Htz1.
Deletions of three yeast genes, SET2, CDC73, and DST1, involved in transcriptional elongation and/or chromatin metabolism were used in conjunction with genetic array technology to screen approximately 4700 yeast deletions and identify double deletion mutants that produce synthetic growth defects. Of the five deletions interacting genetically with all three starting mutations, one encoded the histone H2A variant Htz1 and three encoded components of a novel 13 protein complex, SWR-C, containing the Snf2 family ATPase, Swr1. The SWR-C also copurified with Htz1 and Bdf1, a TFIID-interacting protein that recognizes acetylated histone tails. Deletions of the genes encoding Htz1 and seven nonessential SWR-C components caused a similar spectrum of synthetic growth defects when combined with deletions of 384 genes involved in transcription, suggesting that Htz1 and SWR-C belong to the same pathway. We show that recruitment of Htz1 to chromatin requires the SWR-C. Moreover, like Htz1 and Bdf1, the SWR-C promotes gene expression near silent heterochromatin.


Transcriptional regulation of the androgen receptor cofactor androgen receptor trapped clone-27.
Cofactors modulate nuclear receptor activity and impact human health and disease, yet surprisingly little is known about their transcriptional regulation. Androgen receptor trapped clone-27 (ART-27) is a cofactor that binds to androgen receptor (AR) amino terminus and modulates AR-dependent transcription. Interestingly, ART-27 displays both a cell type- and developmental stage-specific expression pattern. However, the cis-acting elements and trans-acting factors affecting ART-27 gene expression have not been elucidated. We found that ART-27 gene expression is repressed and its promoter is histone H3-K27 tri-methylated in human embryonic kidney cells, but not prostate cells, and the histone deacetylase inhibitor, trichostatin A, relieves this inhibition. The DNA response elements that control the induction of ART-27 gene expression were also characterized. The major cis-acting element corresponds to a consensus cAMP-responsive element (CRE) and binds the CRE-binding protein (CREB) as shown by EMSA and chromatin immunoprecipitation assays. Furthermore, ART-27 promoter activity is induced upon CREB overexpression. Epidermal growth factor, which activates CREB via phosphorylation, also induces ART-27 expression, whereas a reduction in CREB phosphorylation or expression blocks this induction in prostate cells. In human prostate development, both epithelial and stromal cells express CREB; however, active phosphorylated CREB is restricted to epithelial cells where ART-27 is expressed. Based on these findings, we propose a transcriptional regulatory circuit for the developmental expression of ART-27 that includes repression by chromatin modification through a trichostatin A-sensitive factor and activation upon growth factor stimulation via CREB.


Structure of human thrombospondin: complete amino acid sequence derived from cDNA.
Elucidation of the complete amino acid sequence of TSP has suggested plausible explanations for all of the earlier observations on TSP structure and has already suggested new and interesting avenues of investigation aimed at determining the precise function and mechanism of TSP action as a matrix protein. The potential for Ca++-regulated exposure of the RGDA sequence could represent a new level of control for this important recognition system. We have already shown that the binding of TSP to collagen is modulated by the binding of Ca++ to this region of TSP. That is, high Ca++ results in a lower affinity of TSP for collagen, whereas lower Ca++ concentrations enhance the affinity of this interaction. The highly conserved, although short, 15-residue segment, which is nearly identical to region II of the sporozoite malaria protein, may indicate that TSP interacts with a receptor on liver cells, which the malaria parasite uses to gain its initial toehold in the body. If so, this would be another example of pathogenic organisms using a preexisting host recognition system to gain entry to cells where it can multiply. The collagen propeptide-like segment occurs in the collagen-binding domain of TSP and thus may represent the site at which TSP interacts with the collagens. These speculations form the starting point for many exciting lines of experimentation, which will provide us with a better understanding of the role of TSP in hemostasis, in the matrix of a variety of cells, and in development.


Quantitative analysis of organelle distribution and dynamics in Physcomitrella patens protonemal cells.
In the last decade, the moss Physcomitrella patens has emerged as a powerful plant model system, amenable for genetic manipulations not possible in any other plant. This moss is particularly well suited for plant polarized cell growth studies, as in its protonemal phase, expansion is restricted to the tip of its cells. Based on pollen tube and root hair studies, it is well known that tip growth requires active secretion and high polarization of the cellular components. However, such information is still missing in Physcomitrella patens. To gain insight into the mechanisms underlying the participation of organelle organization in tip growth, it is essential to determine the distribution and the dynamics of the organelles in moss cells. We used fluorescent protein fusions to visualize and track Golgi dictyosomes, mitochondria, and peroxisomes in live protonemal cells. We also visualized and tracked chloroplasts based on chlorophyll auto-fluorescence. We showed that in protonemata all four organelles are distributed in a gradient from the tip of the apical cell to the base of the sub-apical cell. For example, the density of Golgi dictyosomes is 4.7 and 3.4 times higher at the tip than at the base in caulonemata and chloronemata respectively. While Golgi stacks are concentrated at the extreme tip of the caulonemata, chloroplasts and peroxisomes are totally excluded. Interestingly, caulonemata, which grow faster than chloronemata, also contain significantly more Golgi dictyosomes and fewer chloroplasts than chloronemata. Moreover, the motility analysis revealed that organelles in protonemata move with low persistency and average instantaneous speeds ranging from 29 to 75 nm/s, which are at least three orders of magnitude slower than those of pollen tube or root hair organelles. To our knowledge, this study reports the first quantitative analysis of organelles in Physcomitrella patens and will make possible comparisons of the distribution and dynamics of organelles from different tip growing plant cells, thus enhancing our understanding of the mechanisms of plant polarized cell growth.


Hypericum lanceolatum (Hypericaceae) as a potential source of new anti-malarial agents: a bioassay-guided fractionation of the stem bark.
Malaria is a major public health threat in Africa, and traditional medicine continues to play a key role in its control especially in rural areas. A bioassay-guided fractionation was carried out in order to evaluate the anti-malarial potential and the safety of the methanol extract of the Hypericum lanceolatum stem bark. The anti-plasmodial activity was assayed by the lactate dehydrogenase method (pLDH) against the multidrug-resistant W2mef laboratory strain, and a field isolate (SHF4) of Plasmodium falciparum. Cytotoxicity tests were carried out using the LLC-MK2 monkey kidney epithelial cells. Five compounds were isolated from the most active and least cytotoxic ethylacetate sub-extract: betulinic acid (HLT1), 2,2',5,6'-tetrahydroxybenzophenone (HLT2), 5-hydroxy-3-methoxyxanthone (HLT3), 3-hydroxy-5-methoxyxanthone (HLT4) and HLT0 (yet to be identified). Three of the tested compounds presented significant anti-plasmodial activities (with 50% inhibitory concentration, IC₅₀ < 5 μM), with 5-hydroxy-3-methoxyxanthone exerting the highest activity, followed by HLT0 and betulinic acid. All the compounds with significant anti-plasmodial activity were non-cytotoxic, except betulinic acid which showed a 50% cytotoxic concentration, CC₅₀ of 25 μg/mL. These findings justify the use of H. lanceolatum stem bark as anti-malarial by traditional healers of Western Cameroon, and could constitute a good basis for further studies towards development of new drug candidates or phytomedicines for malaria.


Nitric oxide in different types of hypertension during pregnancy.
1. Serum nitric oxide (NO) levels (determined by its products of oxidation) were assessed in non-pregnant women, normal pregnant women and patients suffering from mild pre-eclampsia (MPE), severe pre-eclampsia (SPE), chronic hypertension (CHT) and CHT with pre-eclampsia (CHT + PE). The levels of NO products were significantly reduced during pregnancy in MPE (P < 0.001), CHT + PE (P < 0.01) and SPE (P < 0.05). Significant reductions of NO products were also observed in puerperium (P < 0.001) in all groups except CHT + PE (P < 0.05). 2. In normal pregnancy, three events were related to NO levels: (1) negative correlations were found between the levels of nitrite (r = -0.73, P = 0.0003), nitrate (r = -0.53, P = 0.017) and the number of weeks of gestation; (2) in the caesarean section group, the levels of NO at puerperium were significantly lower (P < 0.05) than those during pregnancy; and (3) there was a significant reduction in NO levels in the pregnant women carrying male fetuses as compared with female fetuses (P < 0.05). 3. In SPE, the patients with a family history of hypertension had lower levels of NO compared with the patients without such a history (P < 0.05). 4. A negative correlation was observed between systolic blood pressure, diastolic blood pressure and NO levels in MPE (r = -0.62, P = 0.013 and r = -0.68, P = 0.0049 respectively) and SPE (r = -0.72, P = 0.004 and r = -0.53, P = 0.037 respectively). 5. In SPE, positive correlations were observed between platelet count and nitrite (r = 0.67, P = 0.006) and nitrate levels (r = 0.56, P = 0.028). 6. In MPE, patients with anti-hypertensive treatment showed significantly (P < 0.05) higher levels of NO compared with the non-treated patients. 7. NO may be important in the physiopathology of hypertension during pregnancy, although several factors may affect its levels.


High-throughput glycomic analyses reveal unique oligosaccharide profiles of canine and feline milk samples.
Oligosaccharides are important components of milk, serving as substrates for the intestinal microbiota, acting as antimicrobials that prevent pathogen colonization, and supporting the developing gastrointestinal immune system of neonates. Nutrient composition of canine and feline milk samples has been described previously, but little is known about the oligosaccharide content. Therefore, the objective of this study was to characterize canine and feline milk samples using a high-throughput glycomics approach. 23 dogs (9 Labrador retriever and 14 Labrador retriever x golden retriever crossbreed) and 6 domestic shorthair cats were recruited to the study. Milk samples were collected by manual expression at time points after parturition. Samples were collected across 2 phases per species, differentiated by maternal diet. Following extraction, oligosaccharide content was determined by liquid chromatography-mass spectrometry (LC-MS). In canine milk samples, 3 structures accounted for over 90% of all oligosaccharides detected across two diet groups. These were 3'-sialyllactose, 6'-sialyllactose, and 2'-fucosyllactose. In feline samples, a more diverse range of oligosaccharides was detected, with up to 16 structures present at relative abundance >1% of the total. Difucosyllactose-N-hexaose b, 3'-sialyllactose and lacto-N-neohexaose were all detected at abundances >10% in feline milk samples. Statistically significant differences (p<0.05) in oligosaccharide abundances were observed between collection time points and between diet groups within species. These data explore the oligosaccharide content of canine and feline maternal milk, representing an opportunity to generate a fundamental understanding of the nutritional needs of new-born puppies and kittens.


Dissociative symptoms in media eyewitnesses of an execution.
The first execution in California since 1976 took place recently in the San Quentin Prison gas chamber. Eighteen journalists were invited as media eyewitnesses. The authors postulated that witnessing this execution was psychologically traumatic and that dissociative and anxiety symptoms would be experienced by the journalists. To investigate the prevalence and specific nature of these symptoms, questionnaires were sent to all the journalists about a month after the execution. The questionnaire contained 17 items assessing dissociative symptoms from the authors' questionnaire of 35 highly intercorrelated acute stress items. Fifteen of 18 of the witnesses returned the questionnaire. Items were endorsed on a scale of 0 ("have not experienced") to 5 ("very often experienced") and analyzed as being dichotomously present or absent. The mean age of the respondents was 37.6 (SD = 8.6) and mean years as a journalist were 15.2 (SD = 9.0). Nine subjects were men and six were women. Journalists witnessing the execution endorsed an average of 5.0 dissociative items, ranging from "I saw, heard, or felt things that were not really there" (endorsed by no one) to "I felt estranged or detached from other people" (endorsed by 60%). This prevalence of reported dissociative symptoms is comparable to that seen among persons who endured the recent Oakland/Berkeley, Calif., firestorm. The experience of being an eyewitness to an execution was associated with the development of dissociative symptoms in several journalists.


Super-resolution microscopy reveals photoreceptor-specific subciliary location and function of ciliopathy-associated protein CEP290.
Mutations in the cilium-associated protein CEP290 cause retinal degeneration as part of multiorgan ciliopathies or as retina-specific diseases. The precise location and the functional roles of CEP290 within cilia and, specifically, the connecting cilia (CC) of photoreceptors, remain unclear. We used super-resolution fluorescence microscopy and electron microscopy to localize CEP290 in the CC and in the primary cilia of cultured cells with subdiffraction resolution and to determine effects of CEP290 deficiency in 3 mutant models. Radially, CEP290 localizes in close proximity to the microtubule doublets in the region between the doublets and the ciliary membrane. Longitudinally, it is distributed throughout the length of the CC whereas it is confined to the very base of primary cilia in human retinal pigment epithelium-1 cells. We found Y-shaped links, ciliary substructures between microtubules and membrane, throughout the length of the CC. Severe CEP290 deficiencies in mouse models did not prevent assembly of cilia or cause obvious mislocalization of ciliary components in early stages of degeneration. There were fewer cilia and no normal outer segments in the mutants, but the Y-shaped links were clearly present. These results point to photoreceptor-specific functions of CEP290 essential for CC maturation and stability following the earliest stages of ciliogenesis.


Androgen receptor driven transcription in molecular apocrine breast cancer is mediated by FoxA1.
Breast cancer is a heterogeneous disease and several distinct subtypes exist based on differential gene expression patterns. Molecular apocrine tumours were recently identified as an additional subgroup, characterised as oestrogen receptor negative and androgen receptor positive (ER- AR+), but with an expression profile resembling ER+ luminal breast cancer. One possible explanation for the apparent incongruity is that ER gene expression programmes could be recapitulated by AR. Using a cell line model of ER- AR+ molecular apocrine tumours (termed MDA-MB-453 cells), we map global AR binding events and find a binding profile that is similar to ER binding in breast cancer cells. We find that AR binding is a near-perfect subset of FoxA1 binding regions, a level of concordance never previously seen with a nuclear receptor. AR functionality is dependent on FoxA1, since silencing of FoxA1 inhibits AR binding, expression of the majority of the molecular apocrine gene signature and growth cell growth. These findings show that AR binds and regulates ER cis-regulatory elements in molecular apocrine tumours, resulting in a transcriptional programme reminiscent of ER-mediated transcription in luminal breast cancers.


One-Cell Doubling Evaluation by Living Arrays of Yeast, ODELAY!
Cell growth is a complex phenotype widely used in systems biology to gauge the impact of genetic and environmental perturbations. Due to the magnitude of genome-wide studies, resolution is often sacrificed in favor of throughput, creating a demand for scalable, time-resolved, quantitative methods of growth assessment. We present ODELAY (One-cell Doubling Evaluation by Living Arrays of Yeast), an automated and scalable growth analysis platform. High measurement density and single-cell resolution provide a powerful tool for large-scale multiparameter growth analysis based on the modeling of microcolony expansion on solid media. Pioneered in yeast but applicable to other colony forming organisms, ODELAY extracts the three key growth parameters (lag time, doubling time, and carrying capacity) that define microcolony expansion from single cells, simultaneously permitting the assessment of population heterogeneity. The utility of ODELAY is illustrated using yeast mutants, revealing a spectrum of phenotypes arising from single and combinatorial growth parameter perturbations.


TOP1 modulation during melanoma progression and in adaptative resistance to BRAF and MEK inhibitors.
In melanomas, therapy resistance can arise due to a combination of genetic, epigenetic and phenotypic mechanisms. Due to its crucial role in DNA supercoil relaxation, TOP1 is often considered an essential chemotherapeutic target in cancer. However, how TOP1 expression and activity might differ in therapy sensitive versus resistant cell types is unknown. Here we show that TOP1 expression is increased in metastatic melanoma and correlates with an invasive gene expression signature. More specifically, TOP1 expression is highest in cells with the lowest expression of MITF, a key regulator of melanoma biology. Notably, TOP1 and DNA Single-Strand Break Repair genes are downregulated in BRAFi- and BRAFi/MEKi-resistant cells and TOP1 inhibition decreases invasion markers only in BRAFi/MEKi-resistant cells. Thus, we show three different phenotypes related to TOP1 levels: i) non-malignant cells with low TOP1 levels; ii) metastatic cells with high TOP1 levels and high invasiveness; and iii) BRAFi- and BRAFi/MEKi-resistant cells with low TOP1 levels and high invasiveness. Together, these results highlight the potential role of TOP1 in melanoma progression and resistance.


Clinical highlights from Amsterdam.
This article contains highlights and a selection of the scientific advances from the Clinical Assembly that were presented at the 2015 European Respiratory Society International Congress in Amsterdam, the Netherlands. The most relevant topics for clinicians will be discussed, covering a wide range of areas including interventional pulmonology, rehabilitation and chronic care, thoracic imaging, diffuse and parenchymal lung diseases, and general practice and primary care. In this comprehensive review, exciting novel data will be discussed and put into perspective.


MicroRNA Profiling in Oesophageal Adenocarcinoma Cell Lines and Patient Serum Samples Reveals a Role for miR-451a in Radiation Resistance.
Many patients with Oesophageal Adenocarcinoma (OAC) do not benefit from chemoradiotherapy treatment due to therapy resistance. To better understand the mechanisms involved in resistance and to find potential biomarkers, we investigated the association of microRNAs, which regulate gene expression, with the response to individual treatments, focusing on radiation. Intrinsic radiation resistance and chemotherapy drug resistance were assessed in eight OAC cell lines, and miRNA expression profiling was performed via TaqMan OpenArray qPCR. miRNAs discovered were either uniquely associated with resistance to radiation, cisplatin, or 5-FU, or were common to two or all three of the treatments. Target mRNA pathway analyses indicated several potential mechanisms of treatment resistance. miRNAs associated with the in vitro treatment responses were then investigated for association with pathologic response to neoadjuvant chemoradiotherapy (nCRT) in pre-treatment serums of patients with OAC. miR-451a was associated uniquely with resistance to radiation treatment in the cell lines, and with the response to nCRT in patient serums. Inhibition of miR-451a in the radiation resistant OAC cell line OE19 increased radiosensitivity (Survival Fraction 73% vs. 87%, p = 0.0003), and altered RNA expression. Pathway analysis of effected small non-coding RNAs and corresponding mRNA targets suggest potential mechanisms of radiation resistance in OAC.


Mouse models of USH1C and DFNB18: phenotypic and molecular analyses of two new spontaneous mutations of the Ush1c gene.
We mapped two new recessive mutations causing circling behavior and deafness to the same region on chromosome 7 and showed they are allelic by complementation analysis. One was named 'deaf circler' (allele symbol dfcr) and the other 'deaf circler 2 Jackson' (allele symbol dfcr-2J). Both were shown to be mutations of the Ush1c gene, the mouse ortholog of the gene responsible for human Usher syndrome type IC and for the non-syndromic deafness disorder DFNB18. The Ush1c gene contains 28 exons, 20 that are constitutive and eight that are alternatively spliced. The dfcr mutation is a 12.8 kb intragenic deletion that eliminates three constitutive and five alternatively spliced exons. The dfcr-2J mutation is a 1 bp deletion in an alternatively spliced exon that creates a transcriptional frame shift, changing 38 amino acid codons before introducing a premature stop codon. Both mutations cause congenital deafness and severe balance deficits due to inner ear dysfunction. The stereocilia of cochlear hair cells are disorganized and splayed in mutant mice, with subsequent degeneration of the hair cells and spiral ganglion cells. Harmonin, the protein encoded by Ush1c, has been shown to bind, by means of its PDZ-domains, with the products of other Usher syndrome genes, including Myo7a, Cdh23 and Sans. The complexes formed by these protein interactions are thought to be essential for maintaining the integrity of hair cell stereocilia. The Ush1c mutant mice described here provide a means to directly investigate these interactions in vivo and to evaluate gene structure-function relationships that affect inner ear and eye phenotypes.


The 2018 Mark Coventry, MD Award: Does a Ceramic Bearing Improve Pain, Function, Wear, or Survivorship of TKA in Patients Younger Than 55 Years of Age? A Randomized Trial.
Methods to reduce the revision rate of total knee prostheses because of wear-related issues are important to examine, particularly because younger patients have a disproportionately high risk of revision. (1) Are long-term Knee Society knee and function scores better in younger patients with an oxidized zirconium (OxZr) TKA compared with those with a cobalt-chrome (CoCr) TKA? (2) Are there differences in radiographic signs of loosening or CT findings of osteolysis between OxZr TKAs and CoCr TKAs? (3) Are there fewer polyethylene wear particles in the OxZr TKA than CoCr TKA? (4) Do the groups differ in terms of survivorship free from revision surgery at 13 years? From April 2003 to January 2007, we enrolled 110 patients younger than 55 years of age in this randomized, double-blind, prospective trial. Each patient served as his or her own control and each received an OxZr femoral component in one knee and a CoCr femoral component in the other. The minimum followup was 10 years (mean, 13 years; range, 10-14 years); two died and nine were lost to followup before that time, leaving 99 patients (198 knees) for analysis. There were 28 men and 71 women with a mean age of 53 ± 6 years (range, 40-55 years). We obtained Knee Society knee scores for each knee, but Knee Society function scores, WOMAC scores, and UCLA activity scores were obtained for each patient preoperatively and at each followup. Additionally, we performed radiographic examination preoperatively and at each followup. At a minimum of 10 years (mean, 13 years) followup, we obtained CT scans in all patients. Polyethylene wear particles in the synovial fluid were analyzed at the final followup using thermogravimetric methods and scanning electron microscopy. Survivorship was ascertained using the Kaplan-Meier calculator. A sample size calculation determined that to detect a difference in the Knee Society knee score of 5 points, assuming a SD of 5 points, with an α = 0.05 and β = 0.80, a total of 90 patients would be needed in each group. At the most recent followup, the mean Knee Society knee scores (92 versus 93 points; p = 0.857), function scores (85 versus 85 points; p = 1.000), WOMAC scores (23 versus 23 points; p = 1.000), UCLA activity scores (6.5 versus 6.5 points; p = 1.000), and range of knee motion (125° versus 127°; p = 0.365) were not different between the two groups. There was no radiographic evidence of loosening and no osteolysis visible on CT scan in either group. The weight of polyethylene wear particles produced at the bearing surface was 0.046 ± 0.010 g in 1 g of synovial fluid in patients with an OxZr femoral component and 0.0448 ± 0.0108 g in patients with a CoCr femoral component (p = 0.583). Kaplan-Meier survivorship free from revision was 97% for the OxZr group (95% confidence interval [CI], 93-100) and 98% for the CoCr group (95% CI, 93-100) at 13 years after surgery (p = 0.918). Given the absence of demonstrated superiority of either the CoCr implant or the OxZr implant, we recommend that surgeons and healthcare systems can reasonably choose the less expensive device for routine use, unless there is some compelling reason in an individual patient to choose one over the other (such as severe, documented metal sensitivity). Level I, therapeutic study.


Boxing-acute complications and late sequelae: from concussion to dementia.
Boxing has received increased public attention and acceptance in recent years. However, this development has not been accompanied by a critical discussion of the early and late health complications. We selectively review recent studies on the acute, subacute, and chronic neuropsychiatric consequences of boxing. Cerebral concussions ("knock-outs") are the most relevant acute consequence of boxing. The number of reported cases of death in the ring seems to have mildly decreased. Subacute neuropsychological deficits appear to last longer than subjective symptoms. The associated molecular changes demonstrate neuronal and glial injury correlated with the number and severity of blows to the head (altered total tau, beta-amyloid, neurofilament light protein, glial fibrillary acidic protein, and neuron-specific enolase). The risk of a punch-drunk syndrome (boxer's dementia, dementia pugilistica) as a late effect of chronic traumatic brain injury is associated with the duration of a boxer's career and with his earlier stamina. There are similarities (e.g. increased risk with ApoE4-polymorphism, beta-amyloid pathology) and differences (more tau pathology in boxers) compared with Alzheimer's disease. Protective gear has led to a remarkable reduction of risks in amateur boxing. Similar measures can also be used in professional boxing, but may decrease the thrill, which does appeal to many supporters.


Lemon balm extract causes potent antihyperglycemic and antihyperlipidemic effects in insulin-resistant obese mice.
Over the last decades polyetiological metabolic diseases such as obesity and type 2 diabetes have emerged as a global epidemic. Efficient strategies for prevention and treatment include dietary intervention and the development of validated nutraceuticals. Safe extracts of edible plants provide a resource of structurally diverse molecules that can effectively interfere with multifactorial diseases. In this study, we describe the application of ethanolic lemon balm (Melissa officinalis) leaves extract for the treatment of insulin-resistance and dyslipidemia in mice. We show that lemon balm extract (LBE) activates the peroxisome proliferator-activated receptors (PPARs), which have key roles in the regulation of whole body glucose and lipid metabolism. Application of LBE (0.6 mg/mL) to human primary adipocytes resulted in specific peroxisome proliferator-activated receptor target gene expression. LBE treatment of insulin-resistant high-fat diet-fed C57BL/6 mice (200 mg/kg/day) for 6 weeks considerably reduced hyperglycemia and insulin resistance, plasma triacylglycerol, nonesterified fatty acids and LDL/VLDL cholesterol levels. Taken together, ethanolic lemon balm extract can potentially be used to prevent or concomitantly treat type 2 diabetes and associated disorders such as dyslipidemia and hypercholesterolemia.


Acid-base imbalances during a 120 km endurance race compared by traditional and simplified strong ion difference methods.
Acid-base disturbances are traditionally assessed using the Henderson-Hasselbach equation. The simplified strong ion approach describes more accurately the complex acid-base and electrolyte abnormalities present in endurance horses. To describe acid-base and electrolytes changes in fit horses competing in a FEI*** 120 km endurance race and to compare the traditional vs. strong ion approaches. Thirty horses were initially enrolled in the study. Venous blood samples were obtained before the race (n = 25), at the second (n = 29; 65.4 km) and third vet-gates (n = 23, 97.4 km) and upon race completion (n = 17). Blood gas analysis was performed to determine pH, PCO(2), PO(2), Na(+), K(+) and iCa(++), and calculate HCO(3)(-), base excess and tCO(2). Packed cell volume and total protein, globulin, albumin, lactate, phosphate, glucose and creatinine concentrations, as well as muscle enzymes activities, were also determined. Calculated variables included strong ion difference (SIDm), strong ion gap (SIG) and nonvolatile buffer concentration (A(tot)). A longitudinal linear model using the general estimating equation methodology was used for statistical analysis. Mild but significant increases in PCO(2), SIDm, lactate, plasma protein, globulins and A(tot), as well as a decrease in potassium concentrations were observed from the second vet-gate to race finish when compared to prerace values (P < 0.05). Using the strong ion approach, 67% samples showed acid-base disturbances vs. 70% when using the traditional method, but their interpretations only matched in 24% of measurements. A complex acid-base imbalance characterised by a mild strong ion alkalosis (hypochloraemia attenuated by hyperlactataemia), nonvolatile buffer acidosis and compensatory mild respiratory acidosis were present in most horses, although pH did not significantly change during a 120 km endurance race. The strong ion approach to interpretation of acid-base balance should be favoured over the traditional approach in endurance horses, given the frequent and complex alterations in PCO(2), SIDm and A(tot) during a race.


Green seaweeds (Ulva fasciata sp.) as nitrogen source for fungal cellulase production.
In developing countries, local enzyme production can help decrease the dependency of imported enzymes for bioconversion of e.g. cellulosic feedstocks, but the use of conventional nitrogen sources contributes significantly to such enzyme production cost. Use of local resources is therefore important to consider. Green seaweeds are marine macroalgae that are rich in nitrogen, but not exploited for their nitrogen content. Cellulase production was accomplished by using cocoa pod husk (CPH) and green seaweed (GS) (Ulva fasciata sp.) as growth substrates for Polyporus ciliatus CBS 366.74 in submerged cultivation. The nitrogen concentration of GS was comparable to that of CPH with 0.6% w/v peptone at 4% w/v substrate concentration. A decline of cellulase activity in peptone supplemented GS growth media indicated nitrogen sufficiency of GS to serve as a potential nitrogen source for the fungal growth and cellulase production. Comparison of enzyme production on CPH growth media supplemented with either GS or peptone based on equivalent carbon to nitrogen ratios was done for two Polyporus strains namely; P. ciliatus CBS 366.74 and P. brumalis CBS 470.77. Peptone could be substituted by up to 0.6% w/v with GS at inclusion levels of 50-100% of substrate concentration to attain satisfactory cellulase productivity. However, the cellulase productivity response varied among the two Polyporus species. This study demonstrated that green seaweeds may be used as alternative nitrogen sources for fungal cellulase production.


Physical activity and aging: a life-long story.
The benefits of physical activity in preventing premature mortality have been established by a large set of epidemiological studies. These benefits have been shown both in middle-aged and elderly individuals. Furthermore, the reduction of acute events such as myocardial infarction observed with higher levels of physical activity together with the increase in disease-free life expectancy among the most active individuals supports physical activity's anti-aging effect. This review highlights two models supporting this effect. The first model describes the path to frailty and the second explains that immobilization is a stressor which triggers stress-responses responsible for many chronic diseases. Aging reduces the physiological reserve and can lead to frailty when this reserve cannot allow an appropriate adaptation of the aging body to environmental challenges. The components of this physiological reserve can easily be measured by cardiorespiratory testing. Among them are heart rate reserve and VO(2max), the maximal body oxygen consumption. The opposite effects of exercise training and aging on the physiological reserve are detailed. Underlying mechanisms of both exercise training and aging are described. Sedentary lifestyle accelerates the effects of aging in susceptible individuals. Sedentary lifestyle induces mechanisms which lead to risk factors of chronic diseases and, eventually, to premature death. These pathological mechanisms and their consequences constitute the sedentary lifestyle syndrome.


Relationship of ergonomic stressors to birthweight and gestational age.
Women in many occupations are exposed to ergonomic stressors (eg, prolonged standing, physical exertion, and long work weeks). After a consideration of the major methodological problems encountered in studying effects of ergonomic stressors on birthweight and gestational age, epidemiologic studies that have addressed this issue are reviewed. No individual ergonomic stressor has been consistently associated with effects on gestational age. However, most studies which have attempted to look at the cumulative effect of several stressors report that women in the highest exposure category are at higher risk of giving birth prematurely. In contrast, data regarding an association with birthweight are more variable, and firm conclusions cannot be drawn. While future studies that assess exposure to ergonomic stressors with a more comprehensive and quantitative approach are needed, current evidence is sufficient to suggest that clinicians should take ergonomic stressors into account in the assessment of their patients' risk for preterm delivery.


A Culturally Appropriate Educational Intervention Can Improve Self-Care in Hispanic Patients With Heart Failure: A Pilot Randomized Controlled Trial.
Hispanics constitute the largest US ethnic group and have been shown to have more frequent heart failure (HF) hospitalizations than non-Hispanic whites. Disease management programs can reduce HF hospitalizations and mortality by increasing patient self-care, but most programs are limited to patients who speak English. Therefore, we hypothesize that Project Fluido, a culturally appropriate self-care education intervention, will improve self-care behaviors and knowledge in Hispanic patients with HF compared with usual care (UC). Project Fluido (N = 42) was a randomized controlled pilot trial over 3 months. Patients in the experimental group (n = 22) received individualized education in Spanish using the "teach-back" method on the following: high salt foods, when to call the physician, when to report weight gain and the use of diuretics. They also received a nurse-initiated phone call every 2 weeks, a script for calling their physician with increased symptoms, a weight scale and a daily diary to complete. The UC group (n = 20) received a scale and written information. Self-care was measured using the self-care heart failure index and knowledge using teach-back scores. Four knowledge topics were included when using teach-back. Participants' mean age was 57 ± 14 years, 57% (24) were male, 64% (27) had hypertension, 86% (36) were New York Heart Association Class I-III and 65% (26) had HF with reduced ejection fraction. Participant health literacy scores showed poor health literacy in 31% (n = 13) and 67% (n = 28) spoke Spanish only. Household income was reported as < $20,000 in 93% (n = 39). Self-care and knowledge scores significantly improved (P < 0.04 and P < 0.02, respectively) in the intervention group compared to UC. The intervention utilized in Project Fluido was a remarkably effective method to improve self-care and HF knowledge in a group of Spanish-speaking HF patients. This improvement is in spite of low physical function, health literacy, acculturation and economic challenges. In addition, teach-back was an effective teaching strategy to improve HF knowledge. Future work is needed to investigate the relationship between increased self-care knowledge, readmissions, and mortality in Spanish-speaking patients with HF.


Activated Porous Carbon Spheres with Customized Mesopores through Assembly of Diblock Copolymers for Electrochemical Capacitor.
A series of porous carbon spheres with precisely adjustable mesopores (4-16 nm), high specific surface area (SSA, ∼2000 m2 g-1), and submicrometer particle size (∼300 nm) was synthesized through a facile coassembly of diblock polymer micelles with a nontoxic dopamine source and a common postactivation process. The mesopore size can be controlled by the diblock polymer, polystyrene-block-poly(ethylene oxide) (PS-b-PEO) templates, and has an almost linear dependence on the square root of the degree of polymerization of the PS blocks. These advantageous structural properties make the product a promising electrode material for electrochemical capacitors. The electrochemical capacitive performance was studied carefully by using symmetrical cells in a typical organic electrolyte of 1 M tetraethylammonium tetrafluoroborate/acetonitrile (TEA BF4/AN) or in an ionic liquid electrolyte of 1-ethyl-3-methylimidazolium tetrafluoroborate (EMIMBF4), displaying a high specific capacitance of 111 and 170 F g-1 at 1 A g-1, respectively. The impacts of pore size distribution on the capacitance performance were thoroughly investigated. It was revealed that large mesopores and a relatively low ratio of micropores are ideal for realizing high SSA-normalized capacitance. These results provide us with a simple and reliable way to screen future porous carbon materials for electrochemical capacitors and encourage researchers to design porous carbon with high specific surface area, large mesopores, and a moderate proportion of micropores.


Inhibition of lipopolysaccharide synthesis in Agrobacterium tumefaciens and Aeromonas salmonicida.
Lipopolysaccharide (LPS) synthesis was inhibited, new lipid A metabolites accumulated, and growth ceased, when the plant pathogen Agrobacterium tumefaciens and the fish pathogen Aeromonas salmonicida were treated with an antibacterial agent which specifically inhibits CTP:CMP-3-deoxy-manno-octulosonate cytidylyltransferase (CMP-KDO synthase). The new lipid A metabolites were purified by chromatography on DEAE-cellulose and chemically analysed. Metabolites isolated from both bacterial species contained glucosamine and phosphate in a 1:1 molar ratio, and 3-OH-C14:0 was the major fatty acid present (1 mol and 1.4 mol per mol glucosamine for A. tumefaciens and A. salmonicida, respectively). Inhibition of LPS synthesis by CMP-KDO synthase inhibitor had no effect on the initial kinetics of A. tumefaciens attachment to cultured carrot cells, but did inhibit cell aggregation normally induced by bacterial cellulose synthesis. Bacteria treated with inhibitor remained viable and able to synthesize protein at 15% the rate of control cells, indicating that the lack of cellulose-induced aggregation was not due to the inability of bacteria to make protein, but rather the inability to respond normally to the bacterial-plant cell interaction.


Applying community-based participatory research principles and approaches in clinical trials: forging a new model for cancer clinical research.
Although an estimated 20% of adult cancer patients are medically eligible for a cancer treatment clinical trial (CCT), adult trial participation in the U.S. remains under 3%.- Participation rates are even lower among ethnic and racial minorities and the medically underserved, who tend to have higher cancer mortality rates than the population as a whole.- Given persistent cancer health disparities in these populations, cancer clinical trial participation is increasingly an issue of social justice. Community-based participatory research (CBPR) approaches have been repeatedly recommended as a key strategy for increasing and diversifying cancer clinical trial participation and enhancing their relevance and quality. In 2006, Community-Campus Partnership for Health (CCPH) and the Education Network to Advance Cancer Clinical Trials (ENACCT) received funding from the Agency for Healthcare Research and Quality and the National Cancer Institute (NCI), along with industry and nonprofit partners, to develop the first set of national recommendations to employ CBPR approaches in multisite, phase III cancer clinical trials. The Communities as Partners in Cancer Clinical Trials: Changing Research, Practice and Policy final report, developed through a national advisory committee, two stakeholder meetings and a public vetting process, makes more than fifty detailed recommendations to engage communities in specific and meaningful ways throughout the cancer clinical trial process.1 The report is the first to provide specific guidance as to how and why clinical trials should involve communities affected by cancer-from trial design to implementation to dissemination of results. This paper describes the background and rationale for the initiative, the process used to develop and disseminate the report, and the challenges and opportunities for implementing the report's community-based approaches to cancer clinical research.


Nitric oxide modulation of agonist-evoked intracellular Ca2+ release in neurosecretory PC-12 cells: inhibition of phospholipase C activity via cyclic GMP-dependent protein kinase I.
Nitric oxide is a signaling molecule involved in events crucial to neuronal cell function, such as neurotransmitter release, gene transcription, and neurotoxicity, i.e., a number of processes in which a key role appears to be played by increases in intracellular Ca2+ concentration. In the neurosecretory/neuronal cell line PC-12, we have investigated the role of nitric oxide in the modulation of Ca2+ release from intracellular stores elicited by activation of three different receptors coupled to phosphatidyl-inositol-4,5-bisphosphate hydrolysis, i.e., the purinergic P2U, muscarinic M3, and bradykinin B2 receptors. The results obtained show that nitric oxide donors have an inhibitory effect on agonist-evoked Ca2+ release. This effect is not due to nitric oxide-induced modifications of Ca2+ storage, because the total releasable Ca2+ pool, measured as the radioactivity released by thapsigargin and ionomycin in cells loaded at equilibrium with 45Ca2+, was unchanged. In contrast, nitric oxide donors decreased agonist-evoked inositol-1,4,5-trisphosphate generation and total inositol phosphate accumulation. Similarly, nitric oxide inhibited total inositol phosphate accumulation stimulated by either aluminium fluoride or Ca2+. All of these effects were mimicked by the cGMP analogue 8-bromo-cGMP. When cells were incubated with nitric oxide synthase inhibitors, the results observed were opposite those produced by nitric oxide donors. All of the effects of nitric oxide were abolished when cells were treated with the cGMP-dependent protein kinase I inhibitor KT5823. Furthermore, KT5823 mimicked the effects of nitric oxide synthase inhibitors. We conclude that nitric oxide and Ca2+ signaling pathways are interconnected in PC-12 cells. Modulation of inositol phosphate generation and Ca2+ release by nitric oxide appears to be exerted primarily at the level of phospholipase C functioning and to be mediated by the activation of cGMP-dependent protein kinase I.


NIH Funding within Otolaryngology: 2005-2014.
Objective Analyze grants awarded between 2005 and 2014 to otolaryngology departments that appear in the National Institutes of Health (NIH) RePORTER database, summarize characteristics of grant recipients associated with otolaryngology departments as listed in the RePORTER between 2005 and 2014, and identify trends in otolaryngology NIH funding between 2005 and 2014 by topic. Study Design Case series. Setting NIH database inquiry. Subjects Grant recipients. Methods The RePORTER was queried for all grants awarded to otolaryngology departments between 2005 and 2014. All grants classified as new, renewal, or revision were included while duplicates were excluded. Results In total, 475 grants to 51 institutions were categorized by topic and subtopic. Internet searches were conducted for characteristics of 352 principal investigators. Sixty-seven percent of awardees had a PhD, 22% had an MD, and 11% had an MD/PhD. Sex ratios varied by degrees held. Although 31% of all grant recipients were women, this ratio was not seen when recipients were classified by degree type, with 78% of women holding a PhD compared with 55% of men ( P = .0013). Of the award types, 39% were R01s, 15% were R21s, and 10% were R03s. The top 3 represented topics were otology/neurotology (52%), audiology (25%), and head and neck surgery (14%). The mean annual award amount, after adjusting for inflation to 2014 dollars, was $226,495.76, with 72.8% awarded by the National Institute of Deafness and Communication Disorders. Twenty percent of awardees received multiple grants. Conclusion NIH funding in otolaryngology tends to be awarded to those with PhDs studying the hearing sciences, with 1 in 5 having multiple awards. As in other areas of NIH funding, women are underrepresented overall.


Mixed venous O2 saturation: measured by co-oximetry versus calculated from PVO2.
The objectives of our study were (1) to compare mixed venous saturations calculated by a blood gas machine with those measured directly by a co-oximeter; and (2) to compare the sensitivities and specificities of VO2s derived from these values. Charts were retrospectively reviewed of all MICU patients [n = 16] between December 1, 1991 and January 31, 1992, who required pulmonary artery catheters for their usual care and who had hemoglobin saturations of mixed venous blood concurrently measured by both a co-oximeter (Co-Ox Model 482, Instrumentation Lab, Lexington, MA) and a blood gas analyzer (Nova Biomedical StatLab5, Waltham, MA) which uses a variant of the Severinghaus equation to calculate SVO2 from PVO2). Data used at the time of each SVO2 measurement to calculate oxygen consumption (VO2) further was collected. Available for analysis were 118 mixed venous blood samples. Although the SVO2 values had a correlation coefficient of 0.807 (95% confidence interval [CI] 0.736 to 0.861, Fisher's z-transform), when VO2s were calculated, the blood gas analyzer calculated saturations had a sensitivity of only 58.3% and a specificity of 89%, when compared with those calculated using the saturations measured by the co-oximeter. Attempts to mathematically improve upon the Severinghaus equation and upon an additional four regression equations used by other blood gas analyzers resulted in universally worse sensitivity. If SVO2s calculated by a blood gas machine--rather than those co-oximetrically measured--are used to calculate VO2s, 42% of patients with low O2s will be misclassified as normal and 11% of normals will be misclassified as low. This total error appears to be the result of measurement error by the PO2 electrode of the blood gas analyzer and shifts of the oxyhemoglobin dissociation curve, which are not accounted for in the equation that is used to calculate saturation from measured PO2. We were not able to improve mathematically the sensitivity of any of the available regression equations used by blood gas analyzers to calculate SVO2 from PVO2. Therefore, it remains necessary to use co-oximetrically measured saturations when calculating VO2.


Zebrafish Mnx proteins specify one motoneuron subtype and suppress acquisition of interneuron characteristics.
Precise matching between motoneuron subtypes and the muscles they innervate is a prerequisite for normal behavior. Motoneuron subtype identity is specified by the combination of transcription factors expressed by the cell during its differentiation. Here we investigate the roles of Mnx family transcription factors in specifying the subtypes of individually identified zebrafish primary motoneurons. Zebrafish has three Mnx family members. We show that each of them has a distinct and temporally dynamic expression pattern in each primary motoneuron subtype. We also show that two Mnx family members are expressed in identified VeLD interneurons derived from the same progenitor domain that generates primary motoneurons. Surprisingly, we found that Mnx proteins appear unnecessary for differentiation of VeLD interneurons or the CaP motoneuron subtype. Mnx proteins are, however, required for differentiation of the MiP motoneuron subtype. We previously showed that MiPs require two temporally-distinct phases of Islet1 expression for normal development. Here we show that in the absence of Mnx proteins, the later phase of Islet1 expression is initiated but not sustained, and MiPs become hybrids that co-express morphological and molecular features of motoneurons and V2a interneurons. Unexpectedly, these hybrid MiPs often extend CaP-like axons, and some MiPs appear to be entirely transformed to a CaP morphology. Our results suggest that Mnx proteins promote MiP subtype identity by suppressing both interneuron development and CaP axon pathfinding. This is, to our knowledge, the first report of transcription factors that act to distinguish CaP and MiP subtype identities. Our results also suggest that MiP motoneurons are more similar to V2 interneurons than are CaP motoneurons.


Prevalence, type, and correlates of trauma exposure among adolescent men and women in Soweto, South Africa: implications for HIV prevention.
Youth trauma exposure is associated with syndemic HIV risk. We measured lifetime prevalence, type, and correlates of trauma experience by gender among adolescents living in the HIV hyper-endemic setting of Soweto, South Africa. Using data from the Botsha Bophelo Adolescent Health Survey (BBAHS), prevalence of "ever" experiencing a traumatic event among adolescents (aged 14-19) was assessed using a modified Traumatic Event Screening Inventory-Child (TESI-C) scale (19 items, study alpha = 0.63). We assessed self-reported number of potentially traumatic events (PTEs) experienced overall and by gender. Gender-stratified multivariable logistic regression models assessed independent correlates of 'high PTE score' (≥7 PTEs). Overall, 767/830 (92%) participants were included (58% adolescent women). Nearly all (99.7%) reported experiencing at least one PTE. Median PTE was 7 [Q1,Q3: 5-9], with no gender differences (p = 0.19). Adolescent men reported more violent PTEs (e.g., "seen an act of violence in the community") whereas women reported more non-violent HIV/AIDS-related PTEs (e.g., "family member or someone close died of HIV/AIDS"). High PTE score was independently associated with high food insecurity among adolescent men and women (aOR = 2.63, 95%CI = 1.36-5.09; aOR = 2.57, 95%CI = 1.55-4.26, respectively). For men, high PTE score was also associated with older age (aOR = 1.40/year, 95%CI = 1.21-1.63); and recently moving to Soweto (aOR = 2.78, 95%CI = 1.14-6.76). Among women, high PTE score was associated with depression using the CES-D scale (aOR = 2.00, 95%CI = 1.31-3.03,) and inconsistent condom use vs. no sexual experience (aOR = 2.69, 95%CI = 1.66-4.37). Nearly all adolescents in this study experienced trauma, with gendered differences in PTE types and correlates, but not prevalence. Exposure to PTEs were distributed along social and gendered axes. Among adolescent women, associations with depression and inconsistent condom use suggest pathways for HIV risk. HIV prevention interventions targeting adolescents must address the syndemics of trauma and HIV through the scale-up of gender-transformative, youth-centred, trauma-informed integrated HIV and mental health services.


Lack of perceived sleep improvement after 4-month structured exercise programs.
Many middle-aged women experience decreases in their sleep quality during the menopausal transition. Physical activity has been shown to improve sleep, but few randomized, controlled trials investigating such effects in this population exist. In 164 previously low-active middle-aged women (mean age = 49.9, SD = 3.6), using a 4-month randomized, controlled trial, we examined structured exercise in the form of walking or yoga to determine the effects on perceived sleep quality. Participants completed body composition and fitness assessments and a battery of psychological measures, including the Pittsburgh Sleep Quality Index, at the beginning and end of a 4-month randomized, controlled exercise trial with three arms: walking, yoga, and control. A series of mixed-model repeated-measures univariate analyses of covariance did not reveal any statistically significant intervention effects for total sleep quality or any individual sleep-quality domain. The pattern of effect sizes indicated that the walking group tended to experience small improvements in five of the seven assessed sleep-quality domains. Residual changes in menopausal symptoms and depression were associated with residual changes in total sleep quality. However, after controlling for the effects of physical activity, only menopausal symptoms contributed a unique amount of variance in residual change in total sleep quality. In this study, 4-month moderate-intensity walking and low-intensity yoga programs were ineffective in yielding statistically significant improvements in sleep quality. Exercise interventions of longer durations or greater intensity may be needed for such improvements.


GSK2801, a BAZ2/BRD9 Bromodomain Inhibitor, Synergizes with BET Inhibitors to Induce Apoptosis in Triple-Negative Breast Cancer.
Screening of an inhibitor library targeting kinases and epigenetic regulators identified several molecules having antiproliferative synergy with extraterminal domain (BET) bromodomain (BD) inhibitors (JQ1, OTX015) in triple-negative breast cancer (TNBC). GSK2801, an inhibitor of BAZ2A/B BDs, of the imitation switch chromatin remodeling complexes, and BRD9, of the SWI/SNF complex, demonstrated synergy independent of BRD4 control of P-TEFb-mediated pause-release of RNA polymerase II. GSK2801 or RNAi knockdown of BAZ2A/B with JQ1 selectively displaced BRD2 at promoters/enhancers of ETS-regulated genes. Additional displacement of BRD2 from rDNA in the nucleolus coincided with decreased 45S rRNA, revealing a function of BRD2 in regulating RNA polymerase I transcription. In 2D cultures, enhanced displacement of BRD2 from chromatin by combination drug treatment induced senescence. In spheroid cultures, combination treatment induced cleaved caspase-3 and cleaved PARP characteristic of apoptosis in tumor cells. Thus, GSK2801 blocks BRD2-driven transcription in combination with BET inhibitor and induces apoptosis of TNBC. IMPLICATIONS: Synergistic inhibition of BDs encoded in BAZ2A/B, BRD9, and BET proteins induces apoptosis of TNBC by a combinatorial suppression of ribosomal DNA transcription and ETS-regulated genes.


Solute probes of conformational changes in open complex (RPo) formation by Escherichia coli RNA polymerase at the lambdaPR promoter: evidence for unmasking of the active site in the isomerization step and for large-scale coupled folding in the subsequent conversion to RPo.
Transcription initiation is a multistep process involving a series of requisite conformational changes in RNA polymerase (R) and promoter DNA (P) that create the open complex (RP(o)). Here, we use the small solutes urea and glycine betaine (GB) to probe the extent and type of surface area changes in the formation of RP(o) between Esigma(70) RNA polymerase and lambdaP(R) promoter DNA. Effects of urea quantitatively reflect changes in amide surface and are particularly well-suited to detect coupled protein folding events. GB provides a qualitative probe for the exposure or burial of anionic surface. Kinetics of formation and dissociation of RP(o) reveal strikingly large effects of the solutes on the final steps of RP(o) formation: urea dramatically increases the dissociation rate constant k(d), whereas GB decreases the rate of dissociation. Formation of the first kinetically significant intermediate I(1) is disfavored in urea, and moderately favored by GB. GB slows the rate-determining step that converts I(1) to the second kinetically significant intermediate I(2); urea has no effect on this step. The most direct interpretation of these data is that recognition of promoter DNA in I(1) involves only limited conformational changes. Notably, the data support the following hypotheses: (1) the negatively charged N-terminal domain of sigma(70) remains bound in the "jaws" of polymerase in I(1); (2) the subsequent rate-determining isomerization step involves ejecting this domain from the jaws, thereby unmasking the active site; and (3) final conversion to RP(o) involves coupled folding of the mobile downstream clamp of polymerase.


Select heterozygous Keap1 mutations have a dominant-negative effect on wild-type Keap1 in vivo.
Under homeostatic conditions, Keap1 constitutively mediates the proteasomal degradation Nrf2. However, tertiary changes in Keap1 in response to the cellular environment allow for liberation of Nrf2 to transcriptionally regulate downstream cytoprotective genes that aid in cell survival. KEAP1/NRF2 somatic mutations causing constitutive NRF2 activation have been estimated to occur in approximately 25% of human lung tumors, with similar rates believed to exist in other tissue subtypes. As the stoichiometry between Keap1 and Nrf2 is 2:1, we hypothesized that heterozygous Keap1 mutations could suppress wild-type Keap1 (Keap1(WT)) activity by functioning as a dominant-negative protein through heterodimerization of mutant Keap1 (Keap1(mutant)) with Keap1(WT). When Keap1(G430C) or Keap1(G364C) mutants were expressed in lieu of Keap1(WT), premature juvenile mortality was observed. To test the hypothesis of a dominant-negative effect, Keap1-null mice were engineered to coexpress both Keap1(WT) and Keap1(G430C) transgenes, a phenotype analogous to that previously observed from a human lung tumor. These mice were viable, but phenotypically displayed hyperactivation of downstream Nrf2 target genes and moderate esophageal hyperkeratosis. Similar to the transgenic mouse Keap1(G430C) mutant, Keap1(C273&288A) substitutions also diminished Keap1(WT) activity in vivo. To further delineate involvement of the dominant-negative heterodimer, transgenic mice with a deletion or strategic mutation in Broad-complex, Tramtrack and Bric-a-Brac (BTB) domain that disallowed Keap1 dimerization were generated in the presence of Keap1(WT), and failed to induce the dominant-negative effect in vivo. These results thus demonstrate that sequestration of Keap1(WT) into a Keap1(mutant)-Keap1(WT) heterodimer leads to a dominant-negative effect in vivo and gives rise to Nrf2 activation.


Wide-spectrum reconstruction method for a birefringence interference imaging spectrometer.
We present a mathematical method used to determine the spectrum detected by a birefringence interference imaging spectrometer (BIIS). The reconstructed spectrum has good precision over a wide spectral range, 0.4-1.0 microm. This method considers the light intensity as a function of wavelength and avoids the fatal error caused by birefringence effect in the conventional Fourier transform method. The experimental interferogram of the BIIS is processed in this new way, and the interference data and reconstructed spectrum are in good agreement, proving this method to be very exact and useful. Application of this method will greatly improve the instrument performance.


Modulation of Social Cognition via Hallucinogens and "Entactogens".
Social cognition is a fundamental ability in human everyday lives. Deficits in social functioning also represent a core aspect of many psychiatric disorders. Yet, despite its significance, deficits in social cognition skills are insufficiently targeted by current treatments. Hallucinogens and entactogens have been shown to have the potential to modulate social processing. This article reviews the literature on the influence of hallucinogens and entactogens on social processing in controlled experimental studies in humans and elucidates the underlying neurobiological and neuropharmacological mechanisms. Furthermore, it identifies current knowledge gaps and derives implications for hallucinogen-assisted treatment approaches as well as the development of novel medication for trans-diagnostic impairments in social cognition.


Warmer outdoor temperature is associated with worse cognitive status in multiple sclerosis.
Patients with multiple sclerosis (MS) have more clinical exacerbations and T2 lesion activity during warmer weather. The current study is the first to investigate whether outdoor temperature is related to cognitive status across patients with MS (cross-sectional analysis), and whether cognitive status fluctuates with changes in outdoor temperature within patients with MS (longitudinal analysis). For the cross-sectional analysis, 40 patients with MS and 40 healthy control (HC) subjects were recruited throughout the calendar year. Cognitive status (processing speed, memory) and outdoor temperature were recorded for the day of testing. We calculated partial correlations between cognitive status and temperature for patients with MS and HCs, controlling for demographic and disease variables. For the longitudinal analysis, cognitive status and outdoor temperature were recorded at baseline and 6-month follow-up in a separate sample of 45 patients with MS. We calculated the partial correlation between temperature and cognitive status at follow-up, controlling for baseline temperature and cognitive status (i.e., whether temperature changes are related to cognitive changes within patients with MS). Cross-sectionally, warmer temperature was related to worse cognitive status in patients with MS (r(p) = -0.45, p = 0.006), not in HCs (r(p) = 0.00, p = 0.984). Longitudinally, increased outdoor temperature from baseline to follow-up was related to a decline in cognitive status within patients with MS (r(p) = -0.39, p = 0.010). Cognitive status in patients with MS is worse on warmer days, consistent with a previously established link between heat and lesion activity. Our findings have implications for clinical trial planning, treatment, and lifestyle decisions. We discuss cognitive status as a potential marker of quiescent exacerbations.


Design, implementation, and evaluation of a computerized system to communicate with patients with limited native language proficiency in the perioperative period.
Effective communication with patients having limited proficiency in the native language of anesthesia care providers during the perioperative period is often challenging. We describe how we developed, implemented, and evaluated a computerized system to convey frequently used prerecorded phrases related to perioperative anesthesia care in the languages we most often encounter in such patients. Phrases were chosen through a consensus process among anesthesia department members. These included routine sayings used to inform patients about what they should anticipate, what interventions we are performing, and how they can participate. Common questions requiring a "yes" or "no" answer were also identified. We recorded these phrases using native speakers who were both knowledgeable medically and familiar with the culture of the patients to provide accurate translations. We developed a software application that categorically grouped the phrases and allowed care providers to select a phrase and play the associated sound file to the patient and deployed the program on our touchscreen-enabled anesthesia information management system workstations. A convenience sample of obstetrical patients speaking a Chinese dialect with whom the language program was used were asked to complete an anonymous questionnaire, translated into Chinese, about their experience. Ninety-five percent lower confidence limits (LCLs) were calculated for response proportions. We approached 25 parturients with varying levels of English comprehension, and all agreed to use the language program. Each used it throughout her interaction with the anesthesia care providers during labor and delivery, and all patients completed the survey. Acceptance of the process was high, with all patients indicating that they would like to use it again were they to return for another procedure requiring anesthesia. Eighty-eight percent (LCL = 73%) indicated that having instructions in their native language made them feel more relaxed, whereas the experience was neutral in the remainder. Comprehension of the phrases presented was high, with 96% (LCL = 83%) indicating that they understood all instructions. Ninety-six percent (LCL = 83%) of patients indicated that they would be likely to refer friends and family to our institution based on the availability of this device. Although patient safety likely could be improved by use of a communication device such as the one we developed, our study was insufficiently powered to be able to measure this potential improvement. The process we describe should be useful wherever anesthesia care providers are not able to communicate in the same language as their patients.


Characterization of the 47-kilodalton autosomal chronic granulomatous disease protein: tissue-specific expression and transcriptional control by retinoic acid.
A full-length cDNA clone was isolated for the 47-kilodalton (kDa) subunit of the NADPH oxidase system, whose absence is responsible for the most common form of autosomally inherited chronic granulomatous disease (CGD). It encodes a 44.7-kDa polypeptide, which contains two src homology (SH3) domains and several possible sites for phosphorylation by protein kinase C. We speculate that the SH3 domains may interact with the Rap1 protein associated with cytochrome b-245 (M.T. Quinn, C.A. Parkes, L. Walker, S. Orkin, M. Dinauer, and A. Jesaitis, Nature [London] 342:198-200, 1989). An antiserum raised to the predicted C terminus of the protein detects a polypeptide with an apparent molecular mass of 47 kDa in normal neutrophil granulocytes but not in those from patients with autosomal CGD. The antibody has been used to show that the protein associates with the vacuolar membrane and is phosphorylated in response to phorbol ester treatment. Analysis of a number of tissue types and cell lines shows that expression of the gene is confined to phagocytic cells and B lymphocytes. This observation suggests that patients with CGD may also have a defect in lymphocyte function. p47 protein and mRNA levels increase during retinoic acid-induced neutrophil differentiation of HL60 cells. Nuclear run-on transcription assays show that the gene for p47 is induced at the transcriptional level in a cycloheximide-insensitive manner. These data indicate that this gene is a primary target for regulation by retinoic acid.


Emerging trends in Chinese healthcare: the impact of a rising middle class.
In this report, the authors examine a major phenomenon in the Chinese healthcare marketplace: the explosion of a vigorous and demanding middle class and its impact on the future directions the industry should pursue. Little is known about the expectations of the middle class regarding their healthcare needs other than through anecdotal or informal sources. The views of the middle class are shaped by a variety of influences which include exposure through direct personal contact with international healthcare facilities when traveling abroad or indirectly through increased exposure to the entertainment industry with its abundance of hospital and medical dramas. In addition to a general increased international awareness arising from more advanced education, the perspective of the middle class consumer is also shaped by the reality of what is currently available in China and what is realistic to expect. This report addresses this lack of factual data through an extensive survey of middle class consumers in three major cities in China: Beijing, Shanghai and Chengdu. The survey took a practical and pragmatic approach to exploring this issue. No attempt was made in this study to explain why the consumer feels the way they do about their healthcare expectations. The purpose was simply to outline what expectations the middle class have for the healthcare marketplace in China. In some respects the results are not surprising. They are the expectations that people have in any country, any where. They expect greater privacy and dignity in the care-giving process. They want to be more involved in the decisions that are made regarding their care. They would prefer a personal, private physician as opposed to a revolving door of faces they will never see a second time. They rely strongly on family and friends to advise them on their choice of provider. They expect clean, well-maintained facilities, efficient systems and courteous personnel. In other respects, the conclusions are not necessarily expected. They feel strongly that their hospital or provider of care should be located in a residential area. They are willing in some circumstances to pay more for their care in order to meet their expectations but not significantly more. Despite acknowledging that many of the facets of care they seek such as greater respect for privacy and a generally perceived more positive attitude in the care-giving process are found in foreign physicians, middle class consumers do not express a strong preference for foreign physicians but opt instead for Chinese physicians. In conclusion, the results provide an insight into the expectations held by middle class Chinese of their healthcare providers and outlines a direction for future healthcare development.


Well-Being, New Technologies, and Clinical Evidence for Family Physicians.
This special issue contains several articles on well-being. Not surprisingly, many of these articles are specifically about burnout. The evidence shows differences in the rates of burnout between men and women family physicians as well as their responses. Clinical team structure and organizational change also contribute to burnout. What about the electronic medical record? We are also reminded that burnout is an international issue. There are also several articles on how technology is changing the way family physicians practice. Two articles report on issues regarding screening for frequently seen clinical entities, specifically breast cancer and alcohol misuse. There are also articles looking at the cost of medical assistant turnover in practices, the impact of continuity with a provider on the retention of patients in clinical trials, and much more of interest to family physicians.


Patterns of adult male coresidence among young children of adolescent mothers.
This article uses data from the National Longitudinal Survey of Youth to examine the coresidence patterns of children and adult males during the first three years of a child's life, with special attention to the children of adolescent mothers. Overall, the most common experience was for the children to have an adult male present over the full period. However, there were differences by race and the mother's age when she gave birth. For example, 83 percent of white children and 47 percent of black children born to mothers aged 20 or older lived with an adult male during their entire early childhood, while three quarters of white children and fewer than one-third of black children born to mothers younger than 18 had a male present in their household over their first three years. Among both races, children of older mothers were significantly more likely than others to be born into a household where an adult male was present. The stability of male coresidence varied significantly by the mother's age among white children, but not among blacks. Overall, black children experienced more changes in male coresidence than whites. Finally, the likelihood that the adult male would be married to the mother was positively associated with white race and the age of the mother when she gave birth.


Modeling of wave propagation in layered piezoelectric media by a recursive asymptotic method.
In this paper, a simple asymptotic method to compute wave propagation in a multilayered general anisotropic piezoelectric medium is discussed. The method is based on explicit second and higher order asymptotic representations of the transfer and stiffness matrices for a thin piezoelectric layer. Different orders of the asymptotic expansion are obtained using Padé approximation of the transfer matrix exponent. The total transfer and stiffness matrices for thick layers or multilayers are calculated with high precision by subdividing them into thin sublayers and combining recursively the thin layer transfer and stiffness matrices. The rate of convergence to the exact solution is the same for both transfer and stiffness matrices; however, it is shown that the growth rate of the round-off error with the number of recursive operations for the stiffness matrix is twice that for the transfer matrix; and the stiffness matrix method has better performance for a thick layer. To combine the advantages of both methods, a hybrid method which uses the transfer matrix for the thin layer and the stiffness matrix for the thick layer is proposed. It is shown that the hybrid method has the same stability as the stiffness matrix method and the same round-off error as the transfer matrix method. The method converges to the exact transfer/stiffness matrices essentially with the precision of the computer round-off error. To apply the method to a semispace substrate, the substrate was replaced by an artificial perfect matching layer. The computational results for such an equivalent system are identical with those for the actual system. In our computational experiments, we have found that the advantage of the asymptotic method is its simplicity and efficiency.


Random fiber networks with inclusions: The mechanism of reinforcement.
The mechanical behavior of athermal random fiber networks embedding particulate inclusions is studied in this work. Composites in which the filler size is comparable with the mean segment length of the network are considered. Inclusions are randomly distributed in the network at various volume fractions, and cases in which fibers are rigidly bonded to fillers and in which no such bonding is imposed are studied separately. In the presence of inclusions, the small strain modulus increases, while the ability of the network to strain stiffen decreases relative to the unfilled network case. The reinforcement induced by fillers is most pronounced in sparse networks of floppier filaments that deform in the bending-dominated mode in the unfilled state. As the unfilled network density or the bending stiffness of fibers increases, the effect of filling diminishes rapidly. Fillers lead to a transition from the soft, bending-dominated, to the stiffer, stretching-dominated, deformation mode of the network, a transition which is primarily responsible for the observed overall reinforcement. The confinement, i.e., the restriction on network kinematics imposed by fillers, causes this transition. These results provide a justification for the observed difference in reinforcement obtained in sparsely versus densely cross-linked networks at a given filling fraction and provide guidance for the further development of network-based materials.


Phylogenetic analysis of the oriental-Palearctic-Afrotropical members of Anopheles (Culicidae: Diptera) based on nuclear rDNA and mitochondrial DNA characteristics.
The phylogenetic relationships of Anopheles spp. at the junction of Oriental, Palearctic, and Afrotropical regions in the Iranian plateau were investigated using molecular markers. A 711-bp mitochondrial DNA cytochrome oxidase C subunit I (COI) fragment and the entire second internal transcribed spacer (ITS2) region (286-576 bp) of the nuclear ribosomal DNA (rDNA-ITS2) were sequenced from 14 and 28 taxa, respectively. The analyses included 12 species within Anopheles and 4 within the Myzorhynchus Series of the subgenus Anopheles, 8 within Neocellia, 6 within Myzomyia, 3 within Paramyzomyia, and 1 within the Pyretophorus Series of the subgenus Cellia. The congruent tree topologies of both molecular markers strongly supported monophyly of subgenera Anopheles and Cellia. Phylogenetic trees constructed on the basis of ITS2 sequences could accurately categorize all of the series according to the classical taxonomy but could not distinguish Pyretophorus (Anopheles subpictus) from Paramyzomyia Series. Although sequence data of the COI region were available for only 14 species, the inferred trees revealed good classification among the series but could not show the monophyletic relationship of Cellia spp. Except for a few cases, the tree inferred from ITS2 sequences revealed the best classification for the species studied. The molecular data could significantly improve our understanding of the phylogenetic position of the taxa.


Influenza virus-specific antibody-secreting cells in the murine lung during primary influenza virus infection.
An enzyme-linked immunosorbent plaque assay is described which can reliably enumerate influenza virus-specific antibody-secreting cells and exhibits specificity similar to that of the indirect enzyme-linked immunosorbent assay. The assay was used to characterize the development of specific antibody-secreting cells, principally within lung tissue, during primary murine influenza virus infection after intranasal inoculation. Cells secreting influenza virus-specific immunoglobulin M (IgM), IgG, and IgA were detected in greatest numbers in lung tissue, and the data presented indicated that the cells may have originated from specific B-cell precursors in lung tissue which are demonstratable in vitro. At 11 months after infection, cells secreting IgG and IgA were still present in lung tissue. Influenza virus-specific antibody-secreting cells were also detected in spleen tissue and blood. Antibody-secreting cells appeared earlier in spleen than in lung tissue and declined more rapidly in spleen tissue.


Identifying and managing the adverse effects of immune checkpoint blockade.
Immunotherapy has revolutionized the field of oncology. By inhibiting the cytotoxic T-lymphocyte-associated protein (CTLA-4) and programmed death-1 (PD-1) immune checkpoint pathways, multiple studies have demonstrated greatly improved survival in locally advanced and metastatic cancers including melanoma, renal, lung, gastric, and hepatocellular carcinoma. Trials in other malignancies are ongoing, and undoubtedly the number of drugs in this space will grow beyond the six currently approved by the Food and Drug Administration. However, by altering the immune response to fight cancer, a new class of side effects has emerged known as immune-related adverse events (irAEs). These adverse events are due to overactivation of the immune system in almost any organ of the body, and can occur at any point along a patient's treatment course. irAEs such as endocrinopathies (thyroiditis), colitis, and pneumonitis may occur more commonly. However, other organs such as the liver, heart, or brain may also be affected by immune overactivation and any of these side effects may become life threatening. This review presents an approach to promptly recognize and manage these toxicities, to hopefully minimize morbidity and mortality from irAEs.


Biological, biomolecular, and bio-inspired strategies for detection, extraction, and separations of lanthanides and actinides.
Lanthanides and actinides are elements of ever-increasing technological importance in the modern world. However, the similar chemical and physical properties within these groups make purification of individual elements a challenge. Current industrial standards for the extraction, separation, and purification of these metals from natural sources, recycled materials, and industrial waste are inefficient, relying upon harsh conditions, repetitive steps, and ligands with only modest selectivity. Biological, biomolecular, and bio-inspired strategies towards improving these separations and making them more environmentally sustainable have been researched for many years; however, these methods often have insufficient selectivity for practical application. Recent developments in the understanding of how lanthanides are selectively acquired and used by certain bacteria offer the opportunity for a newer, more efficient take on these designs, as well as the possibility for fundamentally new designs and strategies. Herein, we review current cell-based and biomolecular (primarily small-molecule and protein-based) methods for detection, extraction, and separations of f-block elements. We discuss how the increasing knowledge regarding the selective recognition, uptake, trafficking, and storage of these elements in biological systems has informed and will continue to promote development of novel approaches to achieve these ends.


Freezing adhesion molecules in a state of high-avidity binding blocks eosinophil migration.
Leukocyte extravasation is mediated by multiple interactions of adhesive surface structures with ligands on endothelial cells and matrix components. The functional role of beta 1 (CD29) integrins (or very late antigen [VLA] proteins) in eosinophil migration across polycarbonate filters was examined under several in vitro conditions. Eosinophil migration induced by the chemoattractant C5a or platelet-activating factor was fully inhibited by monoclonal antibody (mAb) 8A2, a recently characterized "activating" CD29 mAb. However, inhibition by mAb 8A2 was observed only under filter conditions that best reflected the in vivo situation, i.e., when the eosinophils migrated over filters preincubated with the extracellular matrix (ECM) protein fibronectin (FN), or when the filters were covered with confluent monolayers of cultured human umbilical vein endothelial cells (HUVEC). When bare untreated filters were used, mAb 8A2 had no effect, whereas the C5a-directed movement was prevented by CD18 mAb. Studies with alpha-subunit (CD49)-specific mAbs indicated that the integrins VLA-4 and -5 mediated migration across FN-preincubated filters, and VLA-2, -4, -5, and -6 were involved in eosinophil migration through filters covered with HUVEC. In contrast with the activating CD29 mAb 8A2, a combination of blocking CD49 mAbs or the nonactivating but blocking CD29 mAb AIIB2 failed to inhibit completely eosinophil migration over FN-preincubated or HUVEC-covered filters. mAb 8A2 stimulated binding to FN but not to HUVEC. Moreover, eosinophil migration over FN-preincubated or HUVEC-covered filters was significantly inhibited by anti-connecting segment 1 (CS-1) mAbs, as well as the soluble CS-1 peptide (unlike migration across bare untreated filters). Thus, inhibition of eosinophil migration by mAb 8A2 depended upon the presence of ECM proteins and not upon the presence of HUVEC per se. In conclusion, "freezing" adhesion receptors of the beta 1 integrin family into their high-avidity binding state by the activating CD29 mAb 8A2 results in a complete inhibition of eosinophil migration under physiological conditions. Hence, activation of beta 1 integrin-mediated cell adhesion may represent a new approach to prevent influx of inflammatory cells.


Validity of affect measurements in evaluating symptom reporting in athletes.
Identifying factors that improve the assessment of athletes' psychological functioning is imperative to make proper return-to-play decisions following concussion. Prior research indicates that an individual's affect is related to symptom reporting. The present study examines two novel methods of affect assessment in college athletes at baseline participating in a sports-concussion management program. A total of 256 athletes completed a neuropsychological baseline battery with measurements of psychological symptoms (BDI-Fast Screen, Post-Concussion Symptom Scale, and ImPact Total Symptom Score) and a measure of affective memory bias (the Affective Verbal Learning Test; AVLT). Examiners completed an observation-based rating of affect. Multivariate analysis of variance and χ2 analyses were conducted to examine the effect of affect on symptom reports. Examiners' Affect Ratings were predictive of broad symptom reporting, while the performance based index of affect (Affective Verbal Learning Test, AVLT) was more predictive of depressive symptoms. These findings suggest that performance on the AVLT may be a useful indicator of self-reported depression in a collegiate athlete sample. Additionally, these results demonstrate that examiners' behavioral assessments of affect are important in the assessment of psychological functioning in athletes. Continued work should focus on developing objective measures that are sensitive and valid for the evaluation of outcomes from concussion.


Confronting deep moral disagreement: the President's Council on Bioethics, moral status, and human embryos.
The report of the President's Council on Bioethics, Human Cloning and Human Dignity, addresses the central ethical, political, and policy issue in human embryonic stem cell research: the moral status of extracorporeal human embryos. The Council members were in sharp disagreement on this issue and essentially failed to adequately engage and respectfully acknowledge each others' deepest moral concerns, despite their stated commitment to do so. This essay provides a detailed critique of the two extreme views on the Council (i.e., embryos have full moral status or they have none at all) and then gives theoretical grounding for our judgment about the intermediate moral status of embryos. It also supplies an account of how to address profound moral disagreements in the public arena, especially by way of constructing a middle ground that deliberately pays sincere respect to the views of those with whom it has deep disagreements.


Native Missing Persons Cases Will Not be Solved by Police Alone: The Case for "Missing Persons Advocates".
This study explored the potential role of victim advocacy in Native American missing person cases. Interviews with 25 tribal and non-tribal victim/social service providers were conducted to assess their perspectives on the factors which make Native Americans vulnerable to going missing, the barriers and challenges regarding reporting and investigating missing persons, as well as how victim/social service providers might better support the families of missing persons. Findings suggest that advocates perceive that responding to and offering services for Native families who experience a missing loved one will be extremely difficult because of the intersection of isolation, poverty, and jurisdictional complexities among tribal lands, combined with social service providers and law enforcement officers' lack of resources and training regarding cultural sensitivity. At the same time, advocates suggest that additional training and resources could help overcome many of these barriers and see a role for victim service providers in responding to missing and murdered Native American persons. Implications and suggestions for practice are discussed.


Application of near infrared reflectance spectroscopy to predict meat and meat products quality: A review.
Over the past three decades, near infrared reflectance (NIR) spectroscopy has been proved to be one of the most efficient and advanced tools for the estimation of quality attributes in meat and meat products. This review focuses on the use of NIR spectroscopy to predict different meat properties, considering the literature published mainly in the last decade. Firstly, the potential of NIR to predict chemical composition (crude protein, intramuscular fat, moisture/dry matter, ash, gross energy, myoglobin and collagen), technological parameters (pH value; L*, a*, b* colour values; water holding capacity; Warner-Bratzler and slice shear force) and sensory attributes (colour, shape, marbling, odour, flavour, juiciness, tenderness or firmness) are reviewed. Secondly, the usefulness of NIR for classification into meat quality grades is presented and thirdly its potential application in the industry is shown. The review indicates that NIR showed high potential to predict chemical meat properties and to categorize meat into quality classes. In contrast, NIR showed limited ability for estimating technological and sensory attributes, which may be mainly due to the heterogeneity of the meat samples and their preparation, the low precision of the reference methods and the subjectivity of assessors in taste panels. Hence, future work to standardize sample preparation and increase the accuracy of reference methods is recommended to improve NIR ability to predict those technological and sensory characteristics. In conclusion, the review shows that NIR has a considerable potential to predict simultaneously numerous meat quality criteria.


No apparent effect of surgery for temporal lobe epilepsy on heart rate variability.
Impaired cardiac autonomic function may contribute to the risk of sudden unexpected death in epilepsy (SUDEP). Clinical observations indicate that successful epilepsy surgery is associated with a reduced risk of SUDEP. However, in a previous study we found impaired cardiac control pre-surgically in patients with poor outcome of surgery, indicating an a priori lower risk in responders to epilepsy surgery. We have now examined the effect of surgery on cardiac autonomic control in the same patients. We used 24 h EKG recordings to assess heart rate variability (HRV) by spectral analysis in 21 consecutive patients after temporal lobe epilepsy surgery. The HRV was compared with healthy controls, with pre-surgical HRV in the same patients, and analyzed in relation to seizure control 1 year after surgery. The patients with poor outcome after surgery had significantly lower SD of RR-intervals, total power, very low frequency power and low frequency power than matched healthy controls. The patients with favorable outcome did not differ from the controls, and the postoperative HRV was not different from HRV before surgery in any of the patient groups. We could not demonstrate any effect on HRV of temporal lobe epilepsy surgery in these patients. The observed lower HRV in the poor outcome group was present already before epilepsy surgery as previously reported. Although our results need confirmation in a larger study, the observations suggest that the increased risk of SUDEP in patients failing epilepsy surgery may be due to a common factor predisposing to surgical failure, impaired HRV as well as to an increased risk of SUDEP.