Immune evasion and recognition of the syphilis spirochete in blood and skin of secondary syphilis patients: two immunologically distinct compartments.
The clinical syndrome associated with secondary syphilis (SS) reflects the propensity of Treponema pallidum (Tp) to escape immune recognition while simultaneously inducing inflammation. To better understand the duality of immune evasion and immune recognition in human syphilis, herein we used a combination of flow cytometry, immunohistochemistry (IHC), and transcriptional profiling to study the immune response in the blood and skin of 27 HIV(-) SS patients in relation to spirochetal burdens. Ex vivo opsonophagocytosis assays using human syphilitic sera (HSS) were performed to model spirochete-monocyte/macrophage interactions in vivo. Despite the presence of low-level spirochetemia, as well as immunophenotypic changes suggestive of monocyte activation, we did not detect systemic cytokine production. SS subjects had substantial decreases in circulating DCs and in IFNγ-producing and cytotoxic NK-cells, along with an emergent CD56-/CD16+ NK-cell subset in blood. Skin lesions, which had visible Tp by IHC and substantial amounts of Tp-DNA, had large numbers of macrophages (CD68+), a relative increase in CD8+ T-cells over CD4+ T-cells and were enriched for CD56+ NK-cells. Skin lesions contained transcripts for cytokines (IFN-γ, TNF-α), chemokines (CCL2, CXCL10), macrophage and DC activation markers (CD40, CD86), Fc-mediated phagocytosis receptors (FcγRI, FcγR3), IFN-β and effector molecules associated with CD8 and NK-cell cytotoxic responses. While HSS promoted uptake of Tp in conjunction with monocyte activation, most spirochetes were not internalized. Our findings support the importance of macrophage driven opsonophagocytosis and cell mediated immunity in treponemal clearance, while suggesting that the balance between phagocytic uptake and evasion is influenced by the relative burdens of bacteria in blood and skin and the presence of Tp subpopulations with differential capacities for binding opsonic antibodies. They also bring to light the extent of the systemic innate and adaptive immunologic abnormalities that define the secondary stage of the disease, which in the skin of patients trends towards a T-cell cytolytic response.


Insulin-induced formation of macromolecular complexes involved in activation of cyclic nucleotide phosphodiesterase 3B (PDE3B) and its interaction with PKB.
Fractionation of 3T3-L1 adipocyte membranes revealed that PDE3B (phosphodiesterase 3B) was associated with PM (plasma membrane) and ER (endoplasmic reticulum)/Golgi fractions, that insulin-induced phosphorylation/activation of PDE3B was greater in internal membranes than PM fractions, and that there was no significant translocation of PDE3B between membrane fractions. Insulin also induced formation of large macromolecular complexes, separated during gel filtration (Superose 6 columns) of solubilized membranes, which apparently contain phosphorylated/activated PDE3B and signalling molecules potentially involved in its activation by insulin, e.g. IRS-1 (insulin receptor substrate-1), IRS-2, PI3K p85 [p85-subunit of PI3K (phosphoinositide 3-kinase)], PKB (protein kinase B), HSP-90 (heat-shock protein 90) and 14-3-3. Expression of full-length recombinant FLAG-tagged murine (M) PDE3B and M3BDelta604 (MPDE3B lacking N-terminal 604 amino acids) indicated that the N-terminal region of MPDE3B was necessary for insulin-induced activation and recruitment of PDE3B. siRNA (small interfering RNA) knock-down of PDE3B indicated that PDE3B was not required for formation of insulin-induced complexes. Wortmannin inhibited insulin-induced assembly of macromolecular complexes, as well as phosphorylation/activation of PKB and PDE3B, and their co-immunoprecipitation. Another PI3K inhibitor, LY294002, and the tyrosine kinase inhibitor, Genistein, also inhibited insulin-induced activation of PDE3B and its co-immunoprecipitation with PKB. Confocal microscopy indicated co-localization of PDE3B and PKB. Recombinant MPDE3B co-immunoprecipitated, and co-eluted during Superose 12 chromatography, to a greater extent with recombinant pPKB (phosphorylated/activated PKB) than dephospho-PKB or p-DeltaPKB [pPKB lacking its PH domain (pleckstrin homology domain)]. Truncated recombinant MPDE3B proteins and pPKB did not efficiently co-immunoprecipitate, suggesting that structural determinants for their interaction reside in, or are regulated by, the N-terminal portion of MPDE3B. Recruitment of PDE3B in macromolecular complexes may be critical for regulation of specific cAMP pools and signalling pathways by insulin, e.g. lipolysis.


Axial, Helical, and Planar Chirality in Directly Linked Basket-Handle Porphyrin Arrays.
The single-electron oxidative dimerization of basket-handle porphyrins (BHPs) with different coordinated metal ions [Cu(II), Ni(II), Pd(II), Zn(II)] yielded directly meso-meso linked dimers in excellent yields. The synthetic protocol is suited for coupling substrates with different meso-substituents (H, Br, aryl, alkyl) opposite to the coupling site. Experimental findings concerning reactivities and selectivities were in good agreement with theoretical investigations using ALIE calculations. The dimers, which all are axially chiral, were resolved into their enantiomers by HPLC on a chiral phase. ECD spectra were measured in the stopped-flow mode and compared with results from quantum-chemical ECD calculations to assign the absolute configuration. One directly linked dimer was further oxidized to a fused system, which possessed a stable helical chirality. Its absolute configuration was again assigned by ECD investigations. Furthermore, functionalized BHPs and tetraarylporphyrins were coupled under Suzuki conditions to give dimers and trimers with either β-meso or β-β linkages. Because of the steric shielding of one of the BHP hemispheres, the products were formed with full diastereoselectivity regarding all porphyrin-porphyrin axes. The stereostructures of these arrays were investigated by quantum-chemical calculations (DFT-D3, TD DFT, and sTD DFT), and the absolute configurations were assigned for all chiral representatives.


Evaluation of a chromogenic assay to measure the factor Xa inhibitory activity of unfractionated heparin in canine plasma.
Unfractionated heparin (UFH) has a complex pharmacologic profile that necessitates patient monitoring to prevent inadequate anticoagulation or overdosage and hemorrhage. Factor Xa inhibitory assays (to measure anti-Xa activity) are used to adjust UFH dosage and define safe and effective regimens for specific thrombotic disorders in humans. In this study, the accuracy, linearity, and clinical utility of a chromogenic assay were assessed for monitoring UFH anti-Xa activity in canine plasma samples. A commercial assay (Rotachrom Heparin, Diagnostica Stago, Parsippany, NJ, USA) was used to measure anti-Xa activity in canine plasma samples spiked with different concentrations of UFH. Background absorbance and assay linearity were compared for canine and human plasmas. Percentage recovery of UFH anti-Xa activity and intra- and interassay imprecisions were investigated by multiple measurements of canine plasma to which known amounts of UFH were added. The spiked plasma samples also were used to determine the heparin sensitivity of an activated partial thromboplastin time (aPTT) test. Canine plasma samples were assayed at a higher dilution than were human plasma samples (3:8 versus 4:8) to eliminate higher background anti-Xa activity in canine plasma. Using this modification, the recovery of anti-Xa activity in canine plasma was linear (R2 > .9) at concentrations of 0 - 0.75 U/mL UFH. Intra- and interassay imprecisions for plasma samples containing 0.5 U/mL UFH were <10%, whereas samples containing 0.25 U/mL UFH had imprecisions of 13% and 24%, respectively. The anti-Xa activity range of 0.5 - 0.75 U/mL caused prolongation of aPTTs to 1.5 - 2.5 times the assay mean. Plasma anti-Xa activity of dogs treated with UFH can be accurately monitored using this commercially available chromogenic assay.


[Surgical approaches in excision of tumors of peripheral nerves].
The authors made an analysis of advantages and disadvantages of different approaches for excision of tumors of peripheral nerves on the basis of their experience with treatment of 216 patients.


5-Methylcytosine RNA Methylation in Arabidopsis Thaliana.
5-Methylcytosine (m5C) is a well-characterized DNA modification, and is also predominantly reported in abundant non-coding RNAs in both prokaryotes and eukaryotes. However, the distribution and biological functions of m5C in plant mRNAs remain largely unknown. Here, we report transcriptome-wide profiling of RNA m5C in Arabidopsis thaliana by applying m5C RNA immunoprecipitation followed by a deep-sequencing approach (m5C-RIP-seq). LC-MS/MS and dot blot analyses reveal a dynamic pattern of m5C mRNA modification in various tissues and at different developmental stages. m5C-RIP-seq analysis identified 6045 m5C peaks in 4465 expressed genes in young seedlings. We found that m5C is enriched in coding sequences with two peaks located immediately after start codons and before stop codons, and is associated with mRNAs with low translation activity. We further demonstrated that an RNA (cytosine-5)-methyltransferase, tRNA-specific methyltransferase 4B (TRM4B), exhibits m5C RNA methyltransferase activity. Mutations in TRM4B display defects in root development and decreased m5C peaks. TRM4B affects the transcript levels of the genes involved in root development, which is positively correlated with their mRNA stability and m5C levels. Our results suggest that m5C in mRNA is a new epitranscriptome marker inArabidopsis, and that regulation of this modification is an integral part of gene regulatory networks underlying plant development.


Embryo implantation: biology, evaluation, and enhancement.
Implantation is an essential step in the development of a pregnancy, but often fails in humans. In assisted reproductive technologies, implantation failure continues to impair treatment outcomes, with distressing results for patients and physicians. Morphokinetics, comprehensive chromosome screening, and the analysis of embryo-derived products detectable in spent culture media offer new means of assessing embryo viability. However, all await validation in randomized controlled trials. Genomic, transcriptomic, and secretomic technologies are similarly being exploited to define specific biomarkers of endometrial receptivity with the aim of identifying novel therapeutic interventions. However, to date no single, clinically relevant molecular marker capable of indicating endometrial receptivity has been reported. Recent work continues to describe the key signalling pathways which result in acceptance or rejection of the implanting embryo. In-vitro studies have revealed that the decidualized endometrium plays an important role in natural embryo selection, which could change our understanding of the aetiology and treatment of reproductive failure. Recent developments in analytical techniques have initiated a search for biomarkers of embryo quality and endometrial receptivity, and in-vitro studies have revealed novel roles for the decidualized endometrium as a biosensor of embryo quality.


Identification of functional modules in protein complexes via hyperclique pattern discovery.
Proteins usually do not act isolated in a cell but function within complicated cellular pathways, interacting with other proteins either in pairs or as components of larger complexes. While many protein complexes have been identified by large-scale experimental studies, due to a large number of false-positive interactions existing in current protein complexes 10, it is still difficult to obtain an accurate understanding of functional modules, which encompass groups of proteins involved in common elementary biological function. In this paper, we present a hyperclique pattern discovery approach for extracting functional modules (hyperclique patterns) from protein complexes. A hyperclique pattern is a type of association pattern containing proteins that are highly affiliated with each other. The analysis of hyperclique patterns shows that proteins within the same pattern tend to present in the protein complex together. Also, statistically significant annotations of proteins in a pattern using the Gene Ontology suggest that proteins within the same hyperclique pattern more likely perform the same function and participate in the same biological process. More interestingly, the 3-D structural view of proteins within a hyperclique pattern reveals that these proteins physically interactwith each other. In addition, we show that several hyperclique patterns corresponding to different functions can participate in the same protein complex as independent modules. Finally, we demonstrate that a hyperclique pattern can be involved in different complexes performing different higher-order biological functions, although the pattern corresponds to a specific elementary biological function.


Natural variation in maize aphid resistance is associated with 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one glucoside methyltransferase activity.
Plants differ greatly in their susceptibility to insect herbivory, suggesting both local adaptation and resistance tradeoffs. We used maize (Zea mays) recombinant inbred lines to map a quantitative trait locus (QTL) for the maize leaf aphid (Rhopalosiphum maidis) susceptibility to maize Chromosome 1. Phytochemical analysis revealed that the same locus was also associated with high levels of 2-hydroxy-4,7-dimethoxy-1,4-benzoxazin-3-one glucoside (HDMBOA-Glc) and low levels of 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one glucoside (DIMBOA-Glc). In vitro enzyme assays with candidate genes from the region of the QTL identified three O-methyltransferases (Bx10a-c) that convert DIMBOA-Glc to HDMBOA-Glc. Variation in HDMBOA-Glc production was attributed to a natural CACTA family transposon insertion that inactivates Bx10c in maize lines with low HDMBOA-Glc accumulation. When tested with a population of 26 diverse maize inbred lines, R. maidis produced more progeny on those with high HDMBOA-Glc and low DIMBOA-Glc. Although HDMBOA-Glc was more toxic to R. maidis than DIMBOA-Glc in vitro, BX10c activity and the resulting decline of DIMBOA-Glc upon methylation to HDMBOA-Glc were associated with reduced callose deposition as an aphid defense response in vivo. Thus, a natural transposon insertion appears to mediate an ecologically relevant trade-off between the direct toxicity and defense-inducing properties of maize benzoxazinoids.


Predicting Reading From Behavioral and Neural Measures - A Longitudinal Event-Related Potential Study.
Fluent reading is characterized by fast and effortless decoding of visual and phonological information. Here we used event-related potentials (ERPs) and neuropsychological testing to probe the neurocognitive basis of reading in a sample of children with a wide range of reading skills. We report data of 51 children who were measured at two time points, i.e., at the end of first grade (mean age 7.6 years) and at the end of fourth grade (mean age 10.5 years). The aim of this study was to clarify whether next to behavioral measures also basic unimodal and bimodal neural measures help explaining the variance in the later reading outcome. Specifically, we addressed the question of whether next to the so far investigated unimodal measures of N1 print tuning and mismatch negativity (MMN), a bimodal measure of audiovisual integration (AV) contributes and possibly enhances prediction of the later reading outcome. We found that the largest variance in reading was explained by the behavioral measures of rapid automatized naming (RAN), block design and vocabulary (46%). Furthermore, we demonstrated that both unimodal measures of N1 print tuning (16%) and filtered MMN (7%) predicted reading, suggesting that N1 print tuning at the early stage of reading acquisition is a particularly good predictor of the later reading outcome. Beyond the behavioral measures, the two unimodal neural measures explained 7.2% additional variance in reading, indicating that basic neural measures can improve prediction of the later reading outcome over behavioral measures alone. In this study, the AV congruency effect did not significantly predict reading. It is therefore possible that audiovisual congruency effects reflect higher levels of multisensory integration that may be less important for reading acquisition in the first year of learning to read, and that they may potentially gain on relevance later on.


Clinical Potential of Expanded Noninvasive Prenatal Testing for Detection of Aneuploidies and Microdeletion/Microduplication Syndromes.
We aimed to evaluate the clinical performance of expanded noninvasive prenatal testing (NIPT-Plus) for the detection of aneuploidies and microdeletion/microduplication syndromes. A total of 7177 pregnant women were enrolled in the study from June 2020 to March 2022 at Xijing Hospital, China. Cases with NIPT-Plus-positive results were further confirmed by chromosomal karyotyping and a chromosomal microarray analysis. A total of 112 positive cases (1.56%) were identified by NIPT-Plus, including 60 chromosome aneuploidies and 52 microdeletion/microduplication syndromes. Ninety-five cases were validated by amniocentesis, and 57 were confirmed with true-positive results, comprising 18 trisomy 21, 4 trisomy 18, 1 trisomy 13, 17 sex chromosome aneuploidies, 1 other aneuploidy, and 16 microdeletion/microduplication syndromes. The positive predictive value of total chromosomal abnormalities was 60% (57/95). For trisomy 21, trisomy 18, trisomy 13, sex chromosome aneuploidies, other aneuploidies and microdeletion/microduplication syndromes, the sensitivity was all 100%, the specificity was 100, 99.986, 100, 99.888, 99.958, and 99.636%, and the positive predictive value was 100, 80, 100, 68, 25, and 38.10%, respectively. For all clinical characteristics, the abnormal maternal serum screening group was found to have the highest prevalence of chromosomal abnormalities (1.54%), and the ultrasound abnormality group presented the highest positive predictive value (73.33%). NIPT-Plus has great potential for the detection of aneuploidies and microdeletion/microduplication syndromes owing to its high sensitivity, safety, and specificity, which greatly reduces unnecessary invasive procedures and the risk of miscarriage and allows informed maternal choice.


Experimental and Computational Study of Ductile Fracture in Small Punch Tests.
A unified experimental-computational study on ductile fracture initiation and propagation during small punch testing is presented. Tests are carried out at room temperature with unnotched disks of different thicknesses where large-scale yielding prevails. In thinner specimens, the fracture occurs with severe necking under membrane tension, whereas for thicker ones a through thickness shearing mode prevails changing the crack orientation relative to the loading direction. Computational studies involve finite element simulations using a shear modified Gurson-Tvergaard-Needleman porous plasticity model with an integral-type nonlocal formulation. The predicted punch load-displacement curves and deformed profiles are in good agreement with the experimental results.


Spin- and density-resolved microscopy of antiferromagnetic correlations in Fermi-Hubbard chains.
The repulsive Hubbard Hamiltonian is one of the foundational models describing strongly correlated electrons and is believed to capture essential aspects of high-temperature superconductivity. Ultracold fermions in optical lattices allow for the simulation of the Hubbard Hamiltonian with control over kinetic energy, interactions, and doping. A great challenge is to reach the required low entropy and to observe antiferromagnetic spin correlations beyond nearest neighbors, for which quantum gas microscopes are ideal. Here, we report on the direct, single-site resolved detection of antiferromagnetic correlations extending up to three sites in spin-1/2 Hubbard chains, which requires entropies per particle well below s* = ln(2). The simultaneous detection of spin and density opens the route toward the study of the interplay between magnetic ordering and doping in various dimensions.


Localization, interactions, and the metal-insulator transition.
Recent advances in our understanding of electronic conduction have pointed up deficiencies in traditional thinking. For a metal at a sufficiently low temperature, it is known both theoretically and experimentally that the conventional picture in terms of the Boltzmann theory breaks down. Improved understanding of both electron localization and the effects of electron-electron interactions in a disordered medium has led to experimentally verifiable predictions. These effects have an important influence on the nature of the metal-insulator transition.


Diagnostic utility of cerebral white matter integrity in early Alzheimer's disease.
We compared white matter integrity with brain atrophy in healthy controls and participants with very mild dementia (Clinical Dementia Rating 0 vs. 0.5) from the Brain Aging Project, a longitudinal study of aging and memory at the University of Kansas Medical Center. Structural magnetic resonance imaging and diffusion tensor imaging (DTI) including fractional anisotropy and mean diffusivity were performed on 27 patients with very mild dementia (Clinical Dementia Rating = 0.5) of the Alzheimer's type (DAT), and 32 cognitively normal subjects. Patient groups were compared across 6 volumetric measures and 14 DTI regions of interest. Very mildly demented patients showed expected disease-related patterns of brain atrophy with reductions in whole-brain and hippocampal volumes most prominent. DTI indices of white matter integrity were mixed. Right parahippocampus showed significant but small disease-related reductions in fractional anisotropy. Right parahippocampus and left internal capsule showed greater mean diffusivity in early DAT compared with controls. A series of discriminant analyses demonstrated that gray matter atrophy was a significantly better predictor of dementia status than were DTI indices. Brain atrophy was most strongly related to very mild DAT. Modest disease-related white matter anomalies were present in temporal cortex, and deep white matter had limited discriminatory diagnostic power, probably because of the very mild stage of disease in these participants.


Field expansion for homonymous hemianopia by optically induced peripheral exotropia.
To describe a novel method for prism correction of hemianopia that provides field-of-view expansion in a convenient and functional format and to evaluate initial clinical application. To expand the upper quadrant of the field, a high power prism segment (30-40delta) is placed base-out across the upper part of the spectacle lens, on the side of the loss, at about the level of the limbus. A similar prism segment at the lower part of the lens is used to treat the lower field. The peripheral location of the prisms causes peripheral exotropia. As a result a scene segment as high as the vertical span of the prism is shifted laterally by 15 to 20 degrees relative to the view of the other eye. At the edge of the hemianopic field loss, objects that would fall in the scotoma of one eye are seen through the prism in the other eye, providing a simultaneous awareness of details within the otherwise absent field-of-view. An approach for fitting the system to patients with abnormal binocular vision (strabismus and amblyopia, with or without diplopia) is discussed as well. The effect of the prisms was evaluated in a noncomparative case series (12 patients). The field expansion is provided at any position of lateral gaze, including gaze away from the side of the scotoma. The effect of this technique on field expansion was demonstrated using standard binocular perimetry. Most patients reported substantial improvement in function and in obstacle avoidance. A novel method for the optical treatment of hemianopia was developed and tested. It was found to be effective in expanding the field and helping patients' mobility.


Set-up and preliminary performance of a digital radiology conference system.
To investigate the set-up and preliminary performance of a digital radiology conference system and its characteristics. The imaging system included a dual-screen image-text workstation with a Matrox G450 demonstration dual-head card, two multimedia projectors, a large screen, and a line-pair and grey scale test card. Patient information on radiology information system (RIS) and images on picture archiving and communication system (PACS) were fetched with GE Radworks software based on Intranet and PACS. The fetching velocity and resolution and grey scale of the images were measured. Patient information in the form of texts and images were demonstrated successfully on two screens with Matrox G450 demonstration dual-head card and switched rapidly between text-image two-screen and image two-screen. The image fetching velocity was fast, the resolution was high, and the grey scale was good. The digital conference radiology system is good, and should be popularized.


Alternative modes for health surveillance surveys: an experiment with web, mail, and telephone.
Web and mail surveys as complements to telephone surveys may help resolve concerns about declining participation in telephone surveys for public health surveillance. Little is known, however, about how responses obtained in Web surveys compare with those from mail or telephone surveys. The Behavioral Risk Factor Surveillance System 2003 core interview was conducted in 3 survey modes: Web (n = 1143), mail (n = 836), and telephone (n = 2072). All 3 samples were drawn randomly. We compared respondent demographics and responses to 8 key questions on health conditions and risk behaviors (including asthma, diabetes, obesity, and HIV testing) across the 3 survey modes. Demographic characteristics of mail and Web respondents varied considerably from those interviewed by telephone. The unadjusted prevalence of outcomes varied by survey mode. After adjustment for respondent demographic characteristics, there were still differences among survey modes in several of the health conditions and risk behaviors, although for some of these, the pattern was different for the unadjusted and adjusted results. As health surveys take advantage of new technologies and moved towards mixed-mode designs, researchers need to test for and, if necessary, account for the effect of mode in the estimates they produce.


Effects of siltuximab on the IL-6-induced signaling pathway in ovarian cancer.
To explore potential therapeutic strategies for interrupting the interleukin-6 (IL-6) signaling pathway, we measured IL-6 expression in ovarian cancer tissues, and evaluated the effects of a monoclonal anti-IL-6 antibody; siltuximab (CNTO 328), on levels of IL-6-induced Stat3 phosphorylation, Stat3 nuclear translocation, and Stat3 downstream antiapoptotic genes. We then looked for enhancing paclitaxel sensitivity in multidrug-resistant ovarian cancer cell lines. Expressions of IL-6 in ovarian cancer patient specimens were assessed by immunohistochemistry. Effects of siltuximab on IL-6-induced activation of Stat3 in an ovarian cancer cell line were determined by Western blot and real-time analysis of Stat3 nucleocytoplasmic translocation. Influence of combination of siltuximab and paclitaxel on tumor growth was evaluated in a xenograft mouse mode in vivo. Metastatic and drug-resistant recurrent tumors have significantly higher IL-6 expression when compared with the matched primary tumors. Siltuximab specifically suppressed IL-6-induced Stat3 phosphorylation and Stat3 nuclear translocation. Treatment with siltuximab significantly decreased the levels of Stat3 downstream proteins such as MCL-1, Bcl-X(L), and survivin. Treatment with siltuximab reduced expression of multiple IL-6-induced genes in these cell lines. Furthermore, siltuximab increased the cytotoxic effects of paclitaxel in a paclitaxel resistant ovarian cancer cell line in vitro, but combination therapy with siltuximab did not have a significant effect on paclitaxel resistant tumor growth in vivo. These results show that siltuximab effectively block the IL-6 signaling pathways and IL-6-induced gene expression. Blockage of IL-6 signaling may provide benefits for the treatment of ovarian cancer.


Rapid isolation and detection of Escherichia coli O157:H7 by use of rainbow agar O157 and PCR assay.
This study has evaluated the use of a commercially available Rainbow agar O157 and polymerase chain reaction (PCR) assays for the detection of Shiga-like toxin producing Escherichia coli and to serotype E. coli O157:H7 from raw meat. The Rainbow agar O157 was found to be selective and sensitive for the screening of the E. coli O157 from artificially and naturally contaminated meat samples. Shiga-like toxin producing E. coli were identified with two primer pairs that amplified fragments of the SLT-I (384 bp) and SLT-II (584 bp). E. coli O157:H7 was serotyped with a primer pair specified for the H7 flagellar gene, which amplify specific DNA fragments (625 bp) from all E. coli O157:H7 strains. The use of Rainbow agar O157 described allows for the presumptive isolation of E. coli O157 in 24 hours. Identification and confirmation of the presumptive isolates as E. coli O157:H7 by PCR assays require additional 6-8 hours. The above-mentioned screening and identification procedures should prove to be a very useful method since it allows for the specific detection of E. coli O157:H7.


Polysalt flocculates as a physicochemical approach in the development of controlled-release oral pharmaceuticals.
In the field of pharmaceutical formulation, polysalt flocculates have received relatively little attention in the literature. Accordingly this investigation could be considered an approach towards the development of oral controlled release dosage forms by polysalt flocculates. The polymer selected for this study was sodium carboxymethylcellulose. This was based on the fact that it forms an insoluble complex with aluminium sulphate, which could be of value in controlling the liberation of the entrapped drug. Drug entrapment was effected by polymeric flocculation, induced by the addition of a physiologically inert electrolyte. Results of this study indicated that this approach provided a physicochemical method for controlling drug release from oral pharmaceuticals, that could be claimed to replace the earlier empirical mechanical methods.


A case study of teaching social responsibility to doctoral students in the climate sciences.
The need to make young scientists aware of their social responsibilities is widely acknowledged, although the question of how to actually do it has so far gained limited attention. A 2-day workshop entitled "Prepared for social responsibility?" attended by doctoral students from multiple disciplines in climate science, was targeted at the perceived needs of the participants and employed a format that took them through three stages of ethics education: sensitization, information and empowerment. The workshop aimed at preparing doctoral students to manage ethical dilemmas that emerge when climate science meets the public sphere (e.g., to identify and balance legitimate perspectives on particular types of geo-engineering), and is an example of how to include social responsibility in doctoral education. The paper describes the workshop from the three different perspectives of the authors: the course teacher, the head of the graduate school, and a graduate student. The elements that contributed to the success of the workshop, and thus make it an example to follow, are (1) the involvement of participating students, (2) the introduction of external expertise and role models in climate science, and (3) a workshop design that focused on ethical analyses of examples from the climate sciences.


The association of leg length and offset reconstruction after total hip arthroplasty with clinical outcomes.
Restoring native hip anatomy and biomechanics is important to create a well-functioning hip arthroplasty. This study investigated the association of hip offset and leg length after hip arthroplasty with clinical outcomes, including patient reported outcome measures, the Trendelenburg Test and gait analysis. In 77 patients undergoing primary hip arthroplasty for osteoarthritis (age mean = 65 SD = 11 years; BMI mean = 27 SD = 5 kg/m2), hip offset and leg length discrepancy were measured on anteroposterior radiographs. The Western Ontario & McMaster Universities Osteoarthritis Index, the Trendelenburg Test and gait were assessed preoperatively, and at 3 and 12 months postoperatively. An inertial measurement unit was used to derive biomechanical parameters, including spatiotemporal gait parameters and tilt angles of the pelvis. Relationships between radiographic and functional outcomes were investigated, and subgroups of patients with >15% decreased and increased femoral offset were analysed separately. Patient-reported function scores and clinical tests demonstrated a few significant, weak correlations with radiographic outcomes (Spearman's ρ range = 0.26-0.32; p < 0.05). Undercorrection of femoral offset was associated with lower patient-reported function scores and with more step irregularity as well as step asymmetry during gait. Postoperative leg length inequality was associated with increased frontal plane tilt angle of the pelvis during the Trendelenburg Test and increased sagittal plane motion of the pelvis during gait. Femoral offset subgroups demonstrated no significant differences for patient-reported function scores and outcomes of the Trendelenburg Test and gait analysis. Reduced hip offset and leg length discrepancy following hip arthroplasty seem to be marginally associated with worse clinical outcomes.


[Is vitamin-E supplementation beneficial or harmful?].
Even though the beneficial effects of vitamin E have been experimentally observed, some clinical trials failed to verify a consistent benefit. One reason for this situation has been the difficulty to make comparisons among different studies. There are differences due to subjects, chemical forms of vitamin E, stages of the disease and others. The intake of high doses of vitamin E as a supplement has increased in many countries. Novel studies, have informed that vitamin E not only has antioxidant properties but can acts through precise molecular actions interacting with proteins and enzymes and modulating cellular signaling, transcriptional regulation and apoptosis induction. However, some recent clinical and meta analysis studies have found that daily supplementation with vitamin E 400 IU or higher is associated to increased mortality. In conclusion, a complete understanding of vitamin E actions at the cell does not exist yet and the controversy about its clinical effects is still present. This paper offers current knowledge on the characteristics, metabolism, properties, beneficial effect as well as the potential toxicity of vitamin E.


Walking along chromosomes with super-resolution imaging, contact maps, and integrative modeling.
Chromosome organization is crucial for genome function. Here, we present a method for visualizing chromosomal DNA at super-resolution and then integrating Hi-C data to produce three-dimensional models of chromosome organization. Using the super-resolution microscopy methods of OligoSTORM and OligoDNA-PAINT, we trace 8 megabases of human chromosome 19, visualizing structures ranging in size from a few kilobases to over a megabase. Focusing on chromosomal regions that contribute to compartments, we discover distinct structures that, in spite of considerable variability, can predict whether such regions correspond to active (A-type) or inactive (B-type) compartments. Imaging through the depths of entire nuclei, we capture pairs of homologous regions in diploid cells, obtaining evidence that maternal and paternal homologous regions can be differentially organized. Finally, using restraint-based modeling to integrate imaging and Hi-C data, we implement a method-integrative modeling of genomic regions (IMGR)-to increase the genomic resolution of our traces to 10 kb.


Trauma, PTSD, and substance use disorders: findings from the Australian National Survey of Mental Health and Well-Being.
The aim of the present study was to examine the association between trauma and posttraumatic stress disorder (PTSD) and substance use disorders and to examine the correlates of substance use disorder plus PTSD comorbidity in the Australian general population. Data were collected from a stratified sample of 10,641 participants as part of the Australian National Survey of Mental Health and Well-Being. A modified version of the Composite International Diagnostic Interview was used to determine the presence of DSM-IV anxiety, affective, and substance use disorders and ICD-10 personality disorders. Substance use disorder plus PTSD was experienced by a significant minority of the Australian general population (0.5%). Among those with PTSD, the most common substance use disorder was an alcohol use disorder (24.1%), whereas among those with a substance use disorder, PTSD was most common among individuals with an opioid use disorder (33.2%). Consistent with U.S. clinical literature, individuals with substance use disorder plus PTSD experience significantly poorer physical and mental health and greater disability than those with substance use disorder alone. In contrast, individuals with PTSD alone and those with substance use disorder plus PTSD shared a remarkably similar clinical profile. It is important that individuals entering treatment for substance use disorder or PTSD be assessed for this comorbidity. The addition of either disorder may present complications that need to be considered for the provision of appropriate treatment. Further research is necessary to ascertain which treatments are most effective in treating comorbid substance use disorder plus PTSD.


Cold Modalities With Different Thermodynamic Properties Produce Different Surface and Intramuscular Temperatures.
OBJECTIVE: To compare surface cooling and deep cooling produced by 3 common forms of cryotherapy. DESIGN AND SETTING: We used a 3 x 4 x 4 factorial with repeated measures on measurement depth and treatment. Independent variables were measurement depth (surface, fat + 1 cm, and fat + 2 cm), treatment (ice bag, Wet-Ice, Flex-i-Cold, and control), and treatment order (first, second, third, and fourth). The lowest temperature recorded was the dependent variable. The treatment order was counterbalanced using a Latin square. Data were analyzed with a repeated-measures analysis of variance. SUBJECTS: Fifteen collegiate volunteers who were free of lower extremity abnormalities. MEASUREMENTS: Thigh skin and thigh intramuscular temperatures (1- and 2-cm subadipose) were measured at 30-second intervals both before and during the 30-minute treatments using fine-wire implantable and surface thermocouples. The coldest recorded temperatures were analyzed. RESULTS: Statistical differences were observed for the depth-by-treatment interaction as well as for the depth and treatment main effects. During cold treatments, superficial depths were colder than deeper depths, and all cold treatments were colder than controls at all depths. For the interaction effect at both the skin surface and at 1-cm subadipose, the ice-bag and Wet-Ice treatments were colder than the Flex-i-Cold treatment. For the interaction at 2-cm subadipose, the cold treatments did not differ from each other. Order of treatments did not produce a significant effect. CONCLUSIONS: During a 30-minute cryotherapy treatment, modalities that undergo a phase change caused lower skin and 1-cm intramuscular temperatures than cold modalities that do not possess these properties. These differences were not seen at 2-cm subadipose but may become apparent with longer treatments.


Animal cysticercosis in indigenous Brazilian villages.
Sanitary inspection of beef and pork meat has been the major tool for diagnosing animal cysticercosis and for preventing taeniasis in Brazil. The indigenous villages Jaguapiru and Bororo are located close to the urban area of the municipality of Dourados in the State of Mato Grosso do Sul, Brazil, where precarious basic sanitation conditions are observed. Both cattle and pigs are raised for self-consumption of meat and milk as well as for sale on the external market, generally without official sanitary inspection. In this study, 96 bovine carcasses and 117 pig blood serum samples from animals raised in these indigenous villages were evaluated for the presence of metacestodes by postmortem evaluation, and anti-Taenia sp. antibodies were investigated using the indirect ELISA test. Metacestode forms were observed in 18.7% of the bovine carcasses, and 9.4% of the pig serum samples were positive for anti-Taenia sp. antibodies. The occurrence of animal cysticercosis in the villages may favor the occurrence of this zoonosis in the indigenous populations. Enforcement of proper slaughter and sanitary inspection conditions are urgently needed for controlling the taeniasis-cysticercosis complex among the indigenous population.


Visual integration enhances associative memory equally for young and older adults without reducing hippocampal encoding activation.
The ability to remember associations between previously unrelated pieces of information is often impaired in older adults (Naveh-Benjamin, 2000). Unitization, the process of creating a perceptually or semantically integrated representation that includes both items in an associative pair, attenuates age-related associative deficits (Bastin et al., 2013; Ahmad et al., 2015; Zheng et al., 2015). Compared to non-unitized pairs, unitized pairs may rely less on hippocampally-mediated binding associated with recollection, and more on familiarity-based processes mediated by perirhinal cortex (PRC) and parahippocampal cortex (PHC). While unitization of verbal materials improves associative memory in older adults, less is known about the impact of visual integration. The present study determined whether visual integration improves associative memory in older adults by minimizing the need for hippocampal (HC) recruitment and shifting encoding to non-hippocampal medial temporal structures, such as the PRC and PHC. Young and older adults were presented with a series of objects paired with naturalistic scenes while undergoing fMRI scanning, and were later given an associative memory test. Visual integration was varied by presenting the object either next to the scene (Separated condition) or visually integrated within the scene (Combined condition). Visual integration improved associative memory among young and older adults to a similar degree by increasing the hit rate for intact pairs, but without increasing false alarms for recombined pairs, suggesting enhanced recollection rather than increased reliance on familiarity. Also contrary to expectations, visual integration resulted in increased hippocampal activation in both age groups, along with increases in PRC and PHC activation. Activation in all three MTL regions predicted discrimination performance during the Separated condition in young adults, while only a marginal relationship between PRC activation and performance was observed during the Combined condition. Older adults showed less overall activation in MTL regions compared to young adults, and associative memory performance was most strongly predicted by prefrontal, rather than MTL, activation. We suggest that visual integration benefits both young and older adults similarly, and provides a special case of unitization that may be mediated by recollective, rather than familiarity-based encoding processes.


Repeated Intravenous Administration of Human Neural Stem Cells Producing Choline Acetyltransferase Exerts Anti-Aging Effects in Male F344 Rats.
Major features of aging might be progressive decreases in cognitive function and physical activity, in addition to withered appearance. Previously, we reported that the intracerebroventricular injection of human neural stem cells (NSCs named F3) encoded the choline acetyltransferase gene (F3.ChAT). The cells secreted acetylcholine and growth factors (GFs) and neurotrophic factors (NFs), thereby improving learning and memory function as well as the physical activity of aged animals. In this study, F344 rats (10 months old) were intravenously transplanted with F3 or F3.ChAT NSCs (1 × 106 cells) once a month to the 21st month of age. Their physical activity and cognitive function were investigated, and brain acetylcholine (ACh) and cholinergic and dopaminergic system markers were analyzed. Neuroprotective and neuroregenerative activities of stem cells were also confirmed by analyzing oxidative damages, neuronal skeletal protein, angiogenesis, brain and muscle weights, and proliferating host stem cells. Stem cells markedly improved both cognitive and physical functions, in parallel with the elevation in ACh levels in cerebrospinal fluid and muscles, in which F3.ChAT cells were more effective than F3 parental cells. Stem cell transplantation downregulated CCL11 and recovered GFs and NFs in the brain, leading to restoration of microtubule-associated protein 2 as well as functional markers of cholinergic and dopaminergic systems, along with neovascularization. Stem cells also restored muscular GFs and NFs, resulting in increased angiogenesis and muscle mass. In addition, stem cells enhanced antioxidative capacity, attenuating oxidative damage to the brain and muscles. The results indicate that NSCs encoding ChAT improve cognitive function and physical activity of aging animals by protecting and recovering functions of multiple organs, including cholinergic and dopaminergic systems, as well as muscles from oxidative injuries through secretion of ACh and GFs/NFs, increased antioxidant elements, and enhanced blood flow.


Silymarin reduces profibrogenic cytokines and reverses hepatic fibrosis in chronic murine schistosomiasis.
In chronic schistosomiasis, hepatic fibrosis is linked to the portal hypertension that causes morbidity in Schistosoma mansoni infection. Silymarin (SIL) is a hepatoprotective and antioxidant medicament largely prescribed against liver diseases that has previously been shown to prevent fibrosis during acute murine schistosomiasis. Here we employed silymarin to try to reverse established hepatic fibrosis in chronic schistosomiasis. Silymarin or vehicle was administered to BALB/c mice every 48 h, starting on the 40th (80 days of treatment), 70th (50 days), or 110th (10 days) day postinfection (dpi). All mice were sacrificed and analyzed at 120 dpi. Treatment with silymarin reduced liver weight and granuloma sizes, reduced the increase in alanine aminotransferase and aspartate aminotransferase levels, and reduced the established hepatic fibrosis (assessed by hydroxyproline contents and picrosirius staining). Treatment with silymarin also reduced the levels of interleukin-13 (IL-13) in serum and increased the gamma interferon (IFN-γ)/IL-13 ratio. There was a linear correlation between IL-13 levels in serum and hydroxyproline hepatic content in both infected untreated and SIL-treated mice, with decreased IL-13 levels corresponding to decreased hydroxyproline hepatic contents. Treatment with either SIL or N-acetylcysteine reduced both proliferation of fibroblast cell lines and basal/IL-13-induced production of collagen I, indicating that besides inhibiting IL-13 production during infection, SIL antioxidant properties most likely contribute to inhibition of collagen production downstream of IL-13. These results show that silymarin interferes with fibrogenic cytokines, reduces established fibrosis, and inhibits downstream effects of IL-13 on fibrogenesis, indicating the drug as a safe and cheap treatment to liver fibrotic disease in schistosomiasis.


Self-reported prenatal influenza vaccination and early childhood vaccine series completion.
No studies have examined associations between prenatal vaccination and childhood vaccination. Mothers who refuse influenza vaccinations during pregnancy report similar attitudes and beliefs to those who refuse vaccinations for their children. The objective of this study was to examine the association between self-reported prenatal influenza vaccination and early childhood vaccination. A retrospective cohort study was conducted with existing surveillance data from 4022 mothers who responded to the 2009-2011 Minnesota Pregnancy Risk Assessment Monitoring System survey and child vaccination records from the Minnesota Immunization Information Connection database. The childhood vaccine series outcome included the following vaccines: diphtheria, tetanus, and pertussis; poliovirus; measles, mumps, and rubella; Haemophilus influenzae type b (Hib); hepatitis B; varicella; and pneumococcal conjugate. To evaluate the association between self-reported prenatal influenza vaccination and early childhood vaccination, unadjusted and adjusted logistic regression was used to estimate log odds for childhood vaccination status, while margins post-estimation commands were used to obtain predicted probabilities and risk differences. Vaccine series completion was 10.86% higher (95% confidence interval (CI) 7.33%-14.40%, adjusted and weighted model) in children of mothers who had a prenatal influenza vaccine compared to those who did not. For individual vaccines in the recommended series, risk differences ranged from 7.83% (95% CI 5.37%, 10.30%) for the Hib vaccine to 10.06% (95% CI 7.29%, 12.83%) for the hepatitis B vaccine. Self-reported prenatal influenza vaccination was associated with increased early childhood vaccination. More research is needed to confirm these results and identify potential intervention strategies.


Hepatic carnitine palmitoyltransferase turnover and translation rates in fed, starved, streptozotocin-diabetic and diethylhexyl phthalate-treated rats.
Hepatic carnitine palmitoyltransferase (CPT) turnover was studied in control and in non-ketotic hyperglycaemic streptozotocin-diabetic rats. The degradation constant (kd) and half-life (t1/2) did not appear to be altered by mild diabetes. The hepatic CPT (micrograms/g of liver) was not increased by the mild, non-ketotic, diabetes. However, the total hepatic CPT (micrograms/liver) was 37% greater in the diabetic animals, owing to the increased liver weight. This resulted from a 40% increase in the synthesis constant (ks). Hepatic CPT activity (total detergent-solubilized) and translation rates were measured in fed, starved (48 h), non-ketotic diabetic, ketotic diabetic and diethylhexyl phthalate (DEHP)-treated rats. CPT activity (m units/mg of mitochondrial protein) was not significantly increased with non-ketotic diabetes (44% increase, but non-significant), but was increased approx. 2-fold with starvation and ketotic diabetes, and 3.5-fold with DEHP treatment. CPT expressed as units/liver was increased non-significantly (23%) in non-ketotic and starved rats, similar to the turnover study, but was significantly increased with ketotic diabetes and with DEHP treatment. mRNA-translation activity for CPT was elevated in all states to a somewhat greater extent than was activity. It was concluded that protein synthesis as a product of increased CPT-mRNA translation activity is a major means of long-term regulation.


The safety of calcium-channel blockers.
Calcium-channel blockers are widely used as an effective treatment for hypertension and angina. Several studies have raised questions about their safety, suggesting that calcium-channel blockers can increase the rates of myocardial infarction (MI) and death, particularly in patients with heart disease. Reviews of these studies have uncovered serious methodological shortcomings or have found them restricted to short-acting drugs, frequently at high doses or used inappropriately. One study was based on old data regarding only short-acting nifedipine, which has never been indicated for patients who have suffered an MI or unstable angina. A case-control study of short-acting verapamil, diltiazem, and nifedipine suggested an increased MI rate was confounded by the higher rates of diabetes and preexisting heart disease in the patients treated with calcium-channel blockers. A third study reported significantly decreased survival only in patients taking short-acting nifedipine; in most of the cases reported, blood pressure was not controlled. While these studies alert us to the limitations of short-acting calcium-channel blockers and the necessity of considering side effects such as neurohormonal stimulation, a number of more recent, better-controlled studies have not confirmed increased risk with calcium-channel blockers when appropriately employed. Calcium-channel blockers should still be considered first-line therapy in appropriately selected patients with hypertension or angina.


A novel scaffold long-acting selective estrogen receptor antagonist and degrader with superior preclinical profile against ER+ breast cancer.
Breast cancer is one of the most common malignant tumors in women worldwide, with the majority of cases showing expression of estrogen receptors (ERs). Although drugs targeting ER have significantly improved survival rates in ER-positive patients, drug resistance remains an unmet clinical need. Fulvestrant, which overcomes selective estrogen receptor modulator (SERM) and AI (aromatase inhibitor) resistance, is currently the only long-acting selective estrogen receptor degrader (SERD) approved for both first and second-line settings. However, it fails to achieve satisfactory efficacy due to its poor solubility. Therefore, we designed and synthesized a series of novel scaffold (THC) derivatives, identifying their activities as ER antagonists and degraders. G-5b, the optimal compound, exhibited binding, antagonistic, degradation or anti-proliferative activities comparable to fulvestrant in ER+ wild type and mutants breast cancer cells. Notably, G-5b showed considerably improved stability and solubility. Research into the underlying mechanism indicated that G-5b engaged the proteasome pathway to degrade ER, subsequently inhibiting the ER signaling pathway and leading to the induction of apoptosis and cell cycle arrest events. Furthermore, G-5b displayed superior in vivo pharmacokinetics and pharmacodynamics properties, coupled with a favorable safety profile in the MCF-7 tamoxifen-resistant (MCF-7/TR) tumor xenograft model. Collectively, G-5b has emerged as a highly promising lead compound, offering potent antagonistic and degradation activities, positioning it as a novel long-acting SERD worthy of further refinement and optimization.


Go Red for Women cardiovascular health-screening evaluation: the dichotomy between awareness and perception of cardiovascular risk in the community.
Cardiovascular (CV) disease is the leading cause of death in U.S. women. Although women are becoming more aware of CV disease, there remains a dichotomy between awareness and perceived risk. We sought to characterize this dichotomy in women attracted to a CV health-screening event, in order to improve strategies to reduce CV risk. During CV health events held in Rochester, Minnesota, from 2007 through 2011, 294 women underwent screening for blood chemistry, body mass index, and blood pressure. Participants also completed surveys about knowledge of CV disease, risk factors, and perceived risk. Risk level was determined from the 2011 American Heart Association Effectiveness-Based Guidelines for Women. Among participants (age, mean [standard deviation], 53 [15.3] years; white, 98%), 99% were aware that CV disease is the leading cause of death in women. Almost half (47%) perceived themselves to be at personal risk, although 65% were determined to be "at risk" and 12% to be at "high risk" according to guidelines, while 22% were uncategorized. Of the 228 participants "at risk" or "high risk," 48% and 21%, respectively, did not perceive themselves to be at any risk. Women attracted to CV health events were aware that CV disease is the leading cause of death for women. However, the magnitude of awareness did not directly translate into perception or understanding of actual personal risk. To decrease the rate of CV disease in women, continued efforts are needed to personalize the importance of, and the methods for reducing, CV risk factors.


Musculoskeletal disorders in Russia at the end of the 20th century.
The situation with musculoskeletal disorders in Russia is described. Developments include markedly increased musculoskeletal morbidity in both adults and children. The burden of disease has also increased both from a medical and socioeconomic standpoint, as the numbers of disabled continue to grow. Recent initiatives under the auspices of the Institute of Rheumatology of the Russian Academy of the Medical Sciences, as well as state support for the Bone and Joint Decade, are highlighted.


Migraine and Yawning.
Yawning is considered to be a symptom that reflects dopaminergic activity, although its pathophysiological mechanism is not yet fully understood. Interestingly, repetitive yawning is seen in some patients during migraine attacks. The aim of this cross-sectional study is to investigate the frequency of yawning during migraine attacks and its association with different characteristics of migraine. Patients with migraine with or without aura were evaluated using questionnaires and diaries to determine the characteristics of headache and accompanying symptoms. Repetitive yawning in the premonitory phase and/or during headache were determined. Three hundred and thirty-nine patients were included in the study. One hundred and fifty-four patients reported repetitive yawning (45.4%) during migraine attacks. Repetitive yawning was reported in the 11.2% of the patients in the premonitory phase, 24.2% during headaches, and 10% both in the premonitory phase and during headaches. Migraine with aura (46.8 vs 31.9%; P = .005), accompanying nausea (89.6 vs 75.1%; P = .001), vomiting (48.7 vs 37.8%; P = .044), osmophobia (66.7 vs 52.3%; P = .024), and cutaneous allodynia (58.2 vs 46%; P = .032) were more common in patients with yawning than without. Other dopaminergic-hypothalamic premonitory symptoms (41.6 vs 26.5%; P = .003), especially sleepiness (17.5 vs 5.9%; P = .001), irritability/anxiety (21.4% vs 11.4%; P = .019), nausea/vomiting (10.4 vs 4.3%; P = .03), and changes in appetite (18.2 vs 9.7%; P = .024), were also more frequent in patients with yawning than without. After being adjusted for all other relevant covariates, the odds of repetitive yawning were increased by the presence of nausea (OR 2.88; 95% CI 1.453-5.726; P = .002) and migraine with aura (OR 1.66; 95% CI 1.035-2.671; P = .036). Our results demonstrated that yawning is a common self-reported symptom leading or accompanying migraine attacks and is associated with aura, nausea and/or vomiting, osmophobia, and cutaneous allodynia in patients with migraine. Although yawning is a rather frequently seen behavior, it is a unique and reliable symptom in patients with migraine that may offer an opportunity for early treatment of migraine attacks.


The Escherichia coli peripheral inner membrane proteome.
Biological membranes are essential for cell viability. Their functional characteristics strongly depend on their protein content, which consists of transmembrane (integral) and peripherally associated membrane proteins. Both integral and peripheral inner membrane proteins mediate a plethora of biological processes. Whereas transmembrane proteins have characteristic hydrophobic stretches and can be predicted using bioinformatics approaches, peripheral inner membrane proteins are hydrophilic, exist in equilibria with soluble pools, and carry no discernible membrane targeting signals. We experimentally determined the cytoplasmic peripheral inner membrane proteome of the model organism Escherichia coli using a multidisciplinary approach. Initially, we extensively re-annotated the theoretical proteome regarding subcellular localization using literature searches, manual curation, and multi-combinatorial bioinformatics searches of the available databases. Next we used sequential biochemical fractionations coupled to direct identification of individual proteins and protein complexes using high resolution mass spectrometry. We determined that the proposed cytoplasmic peripheral inner membrane proteome occupies a previously unsuspected ∼19% of the basic E. coli BL21(DE3) proteome, and the detected peripheral inner membrane proteome occupies ∼25% of the estimated expressed proteome of this cell grown in LB medium to mid-log phase. This value might increase when fleeting interactions, not studied here, are taken into account. Several proteins previously regarded as exclusively cytoplasmic bind membranes avidly. Many of these proteins are organized in functional or/and structural oligomeric complexes that bind to the membrane with multiple interactions. Identified proteins cover the full spectrum of biological activities, and more than half of them are essential. Our data suggest that the cytoplasmic proteome displays remarkably dynamic and extensive communication with biological membrane surfaces that we are only beginning to decipher.


Occupational exposure to 60-hertz magnetic fields and risk of breast cancer in women.
We used data from a large population-based case-control study to test the hypothesis that women whose "usual occupation" entailed exposure to higher than background 60-Hz magnetic fields had a higher risk of breast cancer than women without such exposure. Breast cancer cases were identified from four statewide tumor registries, and controls were randomly selected from lists of licensed drivers and Medicare beneficiaries. Information on usual occupation and breast cancer risk factors was obtained by telephone interview. We calculated adjusted odds ratios from logistic regression models for women holding occupations with potential for low, medium, or high magnetic field exposure, compared with background exposure. There was a modest increase in risk for women with potential for high exposure [odds ratio (OR) = 1.43; 95% confidence interval (CI) = 0.99-2.09], and no increase for women with potential for medium (OR = 1.09; 95% CI = 0.83-1.42) or low (OR = 1.02; 95% CI = 0.91-1.15) exposure The risk among premenopausal women in the highest-exposure category was higher (OR = 1.98; 95% CI = 1.04-3.78) than for postmenopausal women (OR = 1.33; 95% CI = 0.82-2.17).


Identification and Characterization of Tumor-Initiating Cells in Multiple Myeloma.
Treatment failures in cancers, including multiple myeloma (MM), are most likely due to the persistence of a minor population of tumor-initiating cells (TICs), which are noncycling or slowly cycling and very drug resistant. Gene expression profiling and real-time quantitative reverse transcription polymerase chain reaction were employed to define genes differentially expressed between the side-population cells, which contain the TICs, and the main population of MM cells derived from 11 MM patient samples. Self-renewal potential was analyzed by clonogenicity and drug resistance of CD24+ MM cells. Flow cytometry (n = 60) and immunofluorescence (n = 66) were applied on MM patient samples to determine CD24 expression. Therapeutic effects of CD24 antibodies were tested in xenograft MM mouse models containing three to six mice per group. CD24 was highly expressed in the side-population cells, and CD24+ MM cells exhibited high expression of induced pluripotent or embryonic stem cell genes. CD24+ MM cells showed increased clonogenicity, drug resistance, and tumorigenicity. Only 10 CD24+ MM cells were required to develop plasmacytomas in mice (n = three of five mice after 27 days). The frequency of CD24+ MM cells was highly variable in primary MM samples, but the average of CD24+ MM cells was 8.3% after chemotherapy and in complete-remission MM samples with persistent minimal residual disease compared with 1.0% CD24+ MM cells in newly diagnosed MM samples (n = 26). MM patients with a high initial percentage of CD24+ MM cells had inferior progression-free survival (hazard ratio [HR] = 3.81, 95% confidence interval [CI] = 5.66 to 18.34, P < .001) and overall survival (HR = 3.87, 95% CI = 16.61 to 34.39, P = .002). A CD24 antibody inhibited MM cell growth and prevented tumor progression in vivo. Our studies demonstrate that CD24+ MM cells maintain the TIC features of self-renewal and drug resistance and provide a target for myeloma therapy.


The circadian stimulus-oscillator model: Improvements to Kronauer's model of the human circadian pacemaker.
Modeling how patterns of light and dark affect circadian phase is important clinically and organizationally (e.g., the military) because circadian disruption can compromise health and performance. Limit-cycle oscillator models in various forms have been used to characterize phase changes to a limited set of light interventions. We approached the analysis of the van der Pol oscillator-based model proposed by Kronauer and colleagues in 1999 and 2000 (Kronauer99) using a well-established framework from experimental psychology whereby the stimulus (S) acts on the organism (O) to produce a response (R). Within that framework, using four independent data sets utilizing calibrated personal light measurements, we conducted a serial analysis of the factors in the Kronauer99 model that could affect prediction accuracy characterized by changes in dim-light melatonin onset. Prediction uncertainty was slightly greater than 1 h for the new data sets using the original Kronauer99 model. The revised model described here reduced prediction uncertainty for these same data sets by roughly half.


Talking about cases in bioethics: the effect of an intensive course on health care professionals.
Educational efforts in bioethics are prevalent, but little is known about their efficacy. Although previous work indicates that courses in bioethics have a demonstrable effect on medical students, it has not examined their effect on health care professionals. In this report, we describe a study designed to investigate the effect of bioethics education on health care professionals. At the Intensive Bioethics Course, a six-day course held annually at Georgetown University, we administered a questionnaire requiring open-ended responses to vignettes both before and after the course. Following the course, respondents defended their responses more carefully and articulated their thoughts more clearly. In addition, after the course respondents seemed to have a more subtle understanding of the relevant issues in the cases and applied theory to these cases more frequently. These findings help to formulate an understanding of the effect of bioethics education on health care professionals.


Cognitive impairment and transcriptomic profile in hippocampus of young mice after multiple neonatal exposures to sevoflurane.
Children with repeated inhalational anesthesia may develop cognitive disorders. This study aimed to investigate the transcriptome-wide response of hippocampus in young mice that had been exposed to multiple sevoflurane in the neonatal period. Mice received 3% sevoflurane for 2 h on postnatal day (PND) 6, 8, and 10, followed by arterial blood gas test on PND 10, behavioral experiments on PND 31-36, and RNA sequencing (RNA-seq) of hippocampus on PND 37. Functional annotation and protein-protein interaction analyses of differentially expressed genes (DEGs) and quantitative reverse transcription polymerase chain reaction (qPCR) were performed. Neonatal sevoflurane exposures induced cognitive and social behavior disorders in young mice. RNA-seq identified a total of 314 DEGs. Several enriched biological processes (ion channels, brain development, learning, and memory) and signaling pathways (oxytocin signaling pathway and glutamatergic, cholinergic, and GABAergic synapses) were highlighted. As hub-proteins, Pten was involved in nervous system development, synapse assembly, learning, memory, and behaviors, Nos3 and Pik3cd in oxytocin signaling pathway, and Cdk16 in exocytosis and phosphorylation. Some top DEGs were validated by qPCR. This study revealed a transcriptome-wide profile in mice hippocampus after multiple neonatal exposures to sevoflurane, promoting better understanding of underlying mechanisms and investigation of preventive strategies.


Isolation and transplantation of autologous circulating endothelial cells into denuded vessels and prosthetic grafts: implications for cell-based vascular therapy.
Blood-borne endothelial cells originating from adult bone marrow were reported previously. These cells have the properties of an endothelial progenitor cell (EPC) and can be mobilized by cytokines and recruited to sites of neovascularization, where they differentiate into mature endothelial cells. Current protocols for isolation of EPCs from peripheral blood rely on enrichment and selection of CD34+ mononuclear cells. In this report, we describe a streamlined method for the isolation and expansion of EPCs from peripheral blood and evaluate their therapeutic potential for autologous cell-based therapy of injured blood vessels and prosthetic grafts. A subset of unfractionated mononuclear cells exhibited the potential to differentiate in vitro into endothelial cells under selective growth conditions. The cells were efficiently transduced ex vivo by a retroviral vector expressing the LacZ reporter gene and could be expanded to yield sufficient numbers for therapeutic applications. Transplantation of these cells into balloon-injured carotid arteries and into bioprosthetic grafts in rabbits led to rapid endothelialization of the denuded vessels and graft segments, resulting in significant reduction in neointima deposition. We conclude that transplantation of EPCs may play a crucial role in reestablishing endothelial integrity in injured vessels, thereby inhibiting neointimal hyperplasia. These findings may have implications for novel and practical cell-based therapies for vascular disease.


The 1999 patterns of care study of radiotherapy in localized prostate carcinoma: a comprehensive survey of prostate brachytherapy in the United States.
The objective of the current study was to provide descriptive information on a representative national sample of patients with prostate carcinoma who were treated with prostate brachytherapy (PB) in calendar year 1999. A random survey was conducted by the Patterns of Care Study in radiation oncology of 59 facilities (1 facility had no eligible patients) that treated patients with prostate carcinoma in 1999 in the United States. A weighted sample size of 36,496 patients with prostate cancer was included in the 1999 survey (unweighted sample size, 554 patients). The main measures were the clinical characteristics of men prior to treatment and the technical characteristics of PB. Patients were classified into three prognostic groups according to T stage, pretreatment prostate specific antigen (PSA) level, and Gleason score. A weighted sample size of 13,293 patients (36%; unweighted sample size, 162 patients) was treated with PB. Compared with a weighted sample size of 23,203 patients (64%; unweighted sample size, 392 patients) was treated with external beam radiotherapy (EB), patients who received PB were significantly younger (mean age: PB group, 67.7 years; EB group, 70.8 years; P = 0.0006). The mean pretreatment PSA level for the PB group was lower compared with the EB group (9.9 ng/mL vs. 13.33 ng/mL; P = 0.0015). The prognostic groupings were more favorable for patients in the PB group compared with patients in EB group (P = 0.0365). The utilization of androgen deprivation therapy (ADT) in the PB group was similar to the utilization of ADT in the EB group (40.4% vs. 51.3%; P = 0.2282). The vast majority of men who were treated with PB received low-dose-rate, permanent sources (89%). Fifty-four percent of men received PB monotherapy (PBM), and the remaining 46% were treated with EB in addition to PB (EBPB). The prognostic groupings were more favorable for patients in the PBM group compared with patients in the EBPB group (P = 0.0037). Of the men who were treated with low-dose-rate PB, 59% were treated with iodine-125 (I-125), and 41% were treated with palladium-103 (Pd-103). I-125 was used more frequently in men who were treated with PBM, and Pd-103 was used more frequently in men who were treated with EBPB. Postimplantation dosimetry was documented in 61.0% of men who were treated with low-dose-rate PB. Computed tomography imaging was used for 46.5% of men. PB was used in 36% of men who were treated with radiotherapy nationally. The mean age of men who were treated with PB was younger than the population of men who were treated with EB alone. Nearly 50% of men who received PB also received EB. EB was used more frequently in men with higher-risk disease. ADT was used in 40% of patients in the PB group. Techniques and prescription doses were consistent with published guidelines.


The anticancer peptide LL-III alters the physico-chemical properties of a model tumor membrane promoting lipid bilayer permeabilization.
LL-III is an anticancer peptide and has the ability to translocate across tumor cell membranes, which indicates that its action mechanism could be non-membranolytic. However, the exact mechanism through which the peptide gains access into the cell cytoplasm is still unknown. Here, we use a plethora of physico-chemical techniques to characterize the interaction of LL-III with liposomes mimicking the lipid matrix of the tumor cell membrane and its effect on the microstructure and thermotropic properties of the membrane. Furthermore, the effect of the presence of Ca2+ cations at physiological concentration was also investigated. For comparison, the interaction of LL-III with liposomes mimicking the normal cell membrane was also studied. Our results show that the peptide selectively interacts with the model tumor cell membrane. This interaction does not disrupt the lipid bilayer but deeply alters its properties by promoting lipid lateral reorganization and increasing membrane permeability. Overall, our data provide a molecular level description of the interaction of the peptide with the model tumor membrane and are fully consistent with the non-membranolytic action mechanism.


DAX-1, an "antitestis" gene.
The DAX-1 gene has been involved in the dosage sensitive sex reversal (DSS) phenotype, a male-to-female sex-reversal syndrome due to the duplication of a small region of human chromosome Xp21. Dax-1 and Sry have been shown to act antagonistically in the mouse system, where increasing expression of the former leads to female development and increasing activity of the latter to male development. Although these data strongly implicate DAX-1 in sex determination, the mouse and human proteins appear to behave differently. Absence of DAX-1 is responsible for adrenal hypoplasia congenita, a human inherited disorder characterized by adrenal insufficiency and hypogonadotropic hypogonadism. Unlike human patients, Dax-1-deficient XY mice have normal levels of corticotropins and adrenal hormones but are sterile. Dax-1-deficient females are fertile. The DAX-1 protein, an unusual member of the nuclear hormone receptor, may act as a transcriptional repressor. It has been shown to both repress transcriptional activators by direct protein-protein interactions and to bind DNA hairpin structures and repress target genes.


Maxillary growth after palatal surgery. An experimental study on dogs.
Maxillary growth after unilateral closure of surgically induced defects in the hard palate in 6- to 8-week old Beagle puppies was studied. The hard palate except for a 4 mm wide strip of bone in the midline and with its overlaying oral mucoperiosteum was removed. On one side the nasal mucoperiosteum was covered with an autogenous full-thickness skin graft (side SGS) and, on the other side the raw surface (side RS) was left for secondary epithelialization. The animals were killed at 47 to 52 weeks of age. Measurements on the dried skulls of the experimental dogs showed no difference in total maxillary length between the two sides of the maxilla. However, the posterior half of the maxilla was longer in 8 dogs on side SGS and the anterior half of the maxilla was in 7 dogs long on side RS. The palatine suture was displaced towards side RS on all dogs, and the height of the nose was in all dogs greater on side SGS. There was a small consistent tendency that the overall growth was more pronounced on the side with the full-thickness skin graft. It is concluded that reducing the amount of scar tissue by covering raw surfaces with an autogenous full-thickness skin graft is one way to reduce maxillary growth impairment after palatal surgery.


Human parvovirus B19 induced apoptotic bodies contain altered self-antigens that are phagocytosed by antigen presenting cells.
Human parvovirus B19 (B19V) from the erythrovirus genus is known to be a pathogenic virus in humans. Prevalence of B19V infection has been reported worldwide in all seasons, with a high incidence in the spring. B19V is responsible for erythema infectiosum (fifth disease) commonly seen in children. Its other clinical presentations include arthralgia, arthritis, transient aplastic crisis, chronic anemia, congenital anemia, and hydrops fetalis. In addition, B19V infection has been reported to trigger autoimmune diseases such as systemic lupus erythematosus and rheumatoid arthritis. However, the mechanisms of B19V participation in autoimmunity are not fully understood. B19V induced chronic disease and persistent infection suggests B19V can serve as a model for viral host interactions and the role of viruses in the pathogenesis of autoimmune diseases. Here we investigate the involvement of B19V in the breakdown of immune tolerance. Previously, we demonstrated that the non-structural protein 1 (NS 1) of B19V induces apoptosis in non-permissive cells lines and that this protein can cleave host DNA as well as form NS1-DNA adducts. Here we provide evidence that through programmed cell death, apoptotic bodies (ApoBods) are generated by B19V NS1 expression in a non-permissive cell line. Characterization of purified ApoBods identified potential self-antigens within them. In particular, signature self-antigens such as Smith, ApoH, DNA, histone H4 and phosphatidylserine associated with autoimmunity were present in these ApoBods. In addition, when purified ApoBods were introduced to differentiated macrophages, recognition, engulfment and uptake occurred. This suggests that B19V can produce a source of self-antigens for immune cell processing. The results support our hypothesis that B19V NS1-DNA adducts, and nucleosomal and lysosomal antigens present in ApoBods created in non-permissive cell lines, are a source of self-antigens.


Retinal Nerve Fiber Layer May Be Better Preserved in MOG-IgG versus AQP4-IgG Optic Neuritis: A Cohort Study.
Optic neuritis (ON) in patients with anti-myelin oligodendrocyte glycoprotein (MOG)-IgG antibodies has been associated with a better clinical outcome than anti-aquaporin 4 (AQP4)- IgG ON. Average retinal nerve fiber layer thickness (RNFL) correlates with visual outcome after ON. The aim of this study was to examine whether anti-MOG-IgG ON is associated with better average RNFL compared to anti-AQP4-IgG ON, and whether this corresponds with a better visual outcome. A retrospective study was done in a consecutive cohort of patients following anti-AQP4-IgG and anti-MOG-IgG ON. A generalized estimating equation (GEE) models analysis was used to compare average RNFL outcomes in ON eyes of patients with MOG-IgG to AQP4-IgG-positive patients, after adjusting for the number of ON events. The final mean visual field defect and visual acuity were compared between ON eyes of MOG-IgG and AQP4-IgG-positive patients. A correlation between average RNFL and visual function was performed in all study eyes. Sixteen patients were analyzed; ten AQP4-IgG-positive and six MOG-IgG-positive. The six patients with MOG-IgG had ten ON events with disc edema, five of which were bilateral. In the AQP4-IgG-positive ON events, 1/10 patients had disc edema. Final average RNFL was significantly better in eyes following MOG-IgG-ON (75.33μm), compared to 63.63μm in AQP4-IgG-ON, after adjusting for the number of ON attacks (GEE, p = 0.023). Mean visual field defects were significantly smaller (GEE, p = 0.046) among MOG-IgG positive ON eyes compared to AQP-IgG positive ON eyes, but last visual acuity did not differ between the groups (GEE, p = 0.153). Among all eyes, average RNFL positively correlated with mean visual field defect (GEE, p = 0.00015) and negatively correlated with final visual acuity (GEE, p = 0.00005). Following ON, RNFL is better preserved in eyes of patients with MOG-IgG antibodies compared to those with AQP4-IgG antibodies, correlating with better visual outcomes.


Neutrophils exhibit an individual response to different oral bacterial biofilms.
Oral innate immunity is led by neutrophils. It is still unclear how their main antimicrobial mechanisms against different biofilms may contribute to balance or dysregulation in the oral cavity. We investigated the capacity of commensal (Streptococcus oralis) and pathogenic (Porphyromonas gingivalis or Aggregatibacter actinomycetemcomitans) monospecies biofilms to induce or to inhibit selected antimicrobial mechanisms of neutrophils. S. oralis induced neutrophil extracellular traps (NETs) formation, reactive oxygen species (ROS) production, and matrix metalloproteinases (MMPs) 8 and 9 secretion. However, these responses were partially reduced in PMA-activated neutrophils indicating a balance-like neutrophil response, which might be important for the maintenance of oral health. P. gingivalis generally induced ROS. Reduced NET formation and significantly decreased MMP secretion were detectable in activated neutrophils highlighting P. gingivalis' nucleolytic and proteolytic activity, which might support bacterial colonization and pathogenesis of periodontitis. In contrast, A. actinomycetemcomitans did not affect the levels of antimicrobial factors in activated neutrophils and induced NET formation, ROS production, and secretion of MMP-8 and -9 in neutrophils alone, which might contribute to tissue destruction and disease progression. In summary, neutrophil responses to biofilms were species-specific and might support either maintenance of oral health or pathogenesis of periodontitis depending on the species.


Why intraspecific trait variation matters in community ecology.
Natural populations consist of phenotypically diverse individuals that exhibit variation in their demographic parameters and intra- and inter-specific interactions. Recent experimental work indicates that such variation can have significant ecological effects. However, ecological models typically disregard this variation and focus instead on trait means and total population density. Under what situations is this simplification appropriate? Why might intraspecific variation alter ecological dynamics? In this review we synthesize recent theory and identify six general mechanisms by which trait variation changes the outcome of ecological interactions. These mechanisms include several direct effects of trait variation per se and indirect effects arising from the role of genetic variation in trait evolution.


Oncogenic mutations in adenomatous polyposis coli (Apc) activate mechanistic target of rapamycin complex 1 (mTORC1) in mice and zebrafish.
Truncating mutations in adenomatous polyposis coli (APC) are strongly linked to colorectal cancers. APC is a negative regulator of the Wnt pathway and constitutive Wnt activation mediated by enhanced Wnt-β-catenin target gene activation is believed to be the predominant mechanism responsible for APC mutant phenotypes. However, recent evidence suggests that additional downstream effectors contribute to APC mutant phenotypes. We previously identified a mechanism in cultured human cells by which APC, acting through glycogen synthase kinase-3 (GSK-3), suppresses mTORC1, a nutrient sensor that regulates cell growth and proliferation. We hypothesized that truncating Apc mutations should activate mTORC1 in vivo and that mTORC1 plays an important role in Apc mutant phenotypes. We find that mTORC1 is strongly activated in apc mutant zebrafish and in intestinal polyps in Apc mutant mice. Furthermore, mTORC1 activation is essential downstream of APC as mTORC1 inhibition partially rescues Apc mutant phenotypes including early lethality, reduced circulation and liver hyperplasia. Importantly, combining mTORC1 and Wnt inhibition rescues defects in morphogenesis of the anterior-posterior axis that are not rescued by inhibition of either pathway alone. These data establish mTORC1 as a crucial, β-catenin independent effector of oncogenic Apc mutations and highlight the importance of mTORC1 regulation by APC during embryonic development. Our findings also suggest a new model of colorectal cancer pathogenesis in which mTORC1 is activated in parallel with Wnt/β-catenin signaling.


Metabolomics of Human Amniotic Fluid and Maternal Plasma during Normal Pregnancy.
Metabolic profiles of amniotic fluid and maternal blood are sources of valuable information about fetus development and can be potentially useful in diagnosis of pregnancy disorders. In this study, we applied 1H NMR-based metabolic profiling to track metabolic changes occurring in amniotic fluid (AF) and plasma (PL) of healthy mothers over the course of pregnancy. AF and PL samples were collected in the 2nd (T2) and 3rd (T3) trimester, prolonged pregnancy (PP) until time of delivery (TD). A multivariate data analysis of both biofluids reviled a metabolic switch-like transition between 2nd and 3rd trimester, which was followed by metabolic stabilization throughout the rest of pregnancy probably reflecting the stabilization of fetal maturation and development. The differences were further tested using univariate statistics at α = 0.001. In plasma the progression from T2 to T3 was related to increasing levels of glycerol, choline and ketone bodies (3-hydroxybutyrate and acetoacetate) while pyruvate concentration was significantly decreased. In amniotic fluid, T2 to T3 transition was associated with decreasing levels of glucose, carnitine, amino acids (valine, leucine, isoleucine, alanine, methionine, tyrosine, and phenylalanine) and increasing levels of creatinine, succinate, pyruvate, choline, N,N-dimethylglycine and urocanate. Lactate to pyruvate ratio was decreased in AF and conversely increased in PL. The results of our study, show that metabolomics profiling can be used to better understand physiological changes of the complex interdependencies of the mother, the placenta and the fetus during pregnancy. In the future, these results might be a useful reference point for analysis of complicated pregnancies.


The Aryl Hydrocarbon Receptor (AhR) as a Drug Target for Cancer Chemotherapy.
The aryl hydrocarbon receptor (AhR) is overexpressed in some patients with different tumor types, and the receptor can be a negative or positive prognostic factor. There is also evidence from both in vivo and in vitro cell culture models that the AhR can exhibit tumor-specific pro-oncogenic and tumor suppressor-like functions and therefore can be treated with AhR antagonists or agonists, respectively. Successful clinical applications of AhR ligands will require the synthesis and development of selective AhR modulators (SAhRMs) with tumor-specific AhR agonist or antagonist activity, and some currently available compounds such as indole-3-carbinol and diindolylmethane-(DIM) and synthetic AhR antagonists are potential drug candidates. There is also evidence that some AhR-active pharmaceuticals, including tranilast, flutamide, hydroxytamoxifen and omeprazole or their derivatives, may be effective AhR-dependent anticancer agents for single or combination cancer chemotherapies for treatment of breast and pancreatic cancers.


[A comparison of the performances between healthy older adults and persons with Alzheimer's disease on the Rey auditory verbal learning test and the Test de rappel libre/rappel indicé 16 items].
The aim of this research was to compare the performances of healthy elderly (n=40) and individuals with Alzheimer's disease (AD, n=40) on the RL/RI 16, a French adaptation of the Free and cued selective reminding test (FCSRT) and on the Rey auditory verbal learning test (RAVLT). These two verbal episodic memory tests are frequently used in clinical practice in French-speaking populations. Results showed that the RAVLT demonstrated a slightly better sensitivity and sensibility than the RL/RI 16. The RAVLT allowed to classify participants of the two groups without any overlap. Moreover, no floor effect was observed in the RAVLT in AD and ceiling effects were less pronounced in normal controls that in the RL/RI 16. Results observed in the RL/RI 16 showed important ceiling effects and a decline in performance on free recall throughout trials in AD patients. Nonetheless, the latter tool was less sensitive to recency effects than the RAVLT and may thus provide a more realistic view of the long-term memory performance of these patients. The semantic cues provided in the RL/RI 16 appeared to increase intrusions in AD whereas the interference list in the RAVLT was the first source of false recognitions in both healthy elderly and AD. In conclusion, this paper demonstrates both the advantages and disadvantages of these two tools in the evaluation of episodic memory in elderly with and without cognitive deficits.


Novel Sources of Wheat Head Blast Resistance in Modern Breeding Lines and Wheat Wild Relatives.
Wheat head blast (WHB), caused by the fungus Magnaporthe oryzae pathotype triticum, is a devastating disease affecting South America and South Asia. Despite 30 years of intensive effort, the 2NVS translocation from Aegilops ventricosa contains the only useful source of resistance to WHB effective against M. oryzae triticum isolates. The objective of this study was to identify non-2NVS sources of resistance to WHB among elite cultivars, breeding lines, landraces, and wild-relative accessions. Over 780 accessions were evaluated under field and greenhouse conditions in Bolivia, greenhouse conditions in Brazil, and at two biosafety level-3 laboratories in the United States. The M. oryzae triticum isolates B-71 (2012), 008 (2015), and 16MoT001 (2016) were used for controlled experiments, while isolate 008 was used for field experiments. Resistant and susceptible checks were included in all experiments. Under field conditions, susceptible spreaders were inoculated at the tillering stage to guarantee sufficient inoculum. Disease incidence and severity were evaluated as the average rating for each 1-m-row plot. Under controlled conditions, heads were inoculated after full emergence and individually rated for percentage of diseased spikelets. The diagnostic marker Ventriup-LN2 was used to test for the presence of the 2NVS translocation. Four non-2NVS spring wheat International Maize and Wheat Improvement Center breeding lines (CM22, CM49, CM52, and CM61) and four wheat wild-relatives (A. tauschii TA10142, TA1624, TA1667, and TA10140) were identified as resistant (<5% of severity) or moderately resistant (5 to <25% severity) to WHB. Experiments conducted at the seedling stage showed little correlation with disease severity at the head stage. M. oryzae triticum isolate 16MoT001 was significantly more aggressive against 2NVS-based varieties. The low frequency of WHB resistance and the increase in aggressiveness of newer M. oryzae triticum isolates highlight the threat that the disease poses to wheat production worldwide and the urgent need to identify and characterize new resistance genes that can be used in breeding for durably resistant varieties.


A mutation linked with Bartter's syndrome locks Kir 1.1a (ROMK1) channels in a closed state.
Mutations in the inward rectifying renal K(+) channel, Kir 1.1a (ROMK), have been linked with Bartter's syndrome, a familial salt-wasting nephropathy. One disease-causing mutation removes the last 60 amino acids (332-391), implicating a previously unappreciated domain, the extreme COOH terminus, as a necessary functional element. Consistent with this hypothesis, truncated channels (Kir 1.1a 331X) are nonfunctional. In the present study, the roles of this domain were systematically evaluated. When coexpressed with wild-type subunits, Kir 1.1a 331X exerted a negative effect, demonstrating that the mutant channel is synthesized and capable of oligomerization. Plasmalemma localization of Kir 1.1a 331X green fluorescent protein (GFP) fusion construct was indistinguishable from the GFP-wild-type channel, demonstrating that mutant channels are expressed on the oocyte plasma membrane in a nonconductive or locked-closed conformation. Incremental reconstruction of the COOH terminus identified amino acids 332-351 as the critical residues for restoring channel activity and uncovered the nature of the functional defect. Mutant channels that are truncated at the extreme boundary of the required domain (Kir 1.1a 351X) display marked inactivation behavior characterized by frequent occupancy in a long-lived closed state. A critical analysis of the Kir 1.1a 331X dominant negative effect suggests a molecular mechanism underlying the aberrant closed-state stabilization. Coexpression of different doses of mutant with wild-type subunits produced an intermediate dominant negative effect, whereas incorporation of a single mutant into a tetrameric concatemer conferred a complete dominant negative effect. This identifies the extreme COOH terminus as an important subunit interaction domain, controlling the efficiency of oligomerization. Collectively, these observations provide a mechanistic basis for the loss of function in one particular Bartter's-causing mutation and identify a structural element that controls open-state occupancy and determines subunit oligomerization. Based on the overlapping functions of this domain, we speculate that intersubunit interactions within the COOH terminus may regulate the energetics of channel opening.


Developing Digital Media to Destigmatize Emergency Department Human Immunodeficiency Virus Testing Among Sexual and Racial Minority Youth: A Hyper-iterative Methodology.
Rates of undiagnosed youth human immunodeficiency virus (HIV) remain problematically high across the United States and internationally. In addition, youth HIV test rates remain consistently low, and youth with HIV remain undiagnosed for longer periods of time as compared with older populations. Youth HIV remains especially persistent among African American adolescents and emerging adults, who are less likely to have consistent access to primary care and thus to HIV testing and prevention education. Therefore, increasing youth HIV test rates has become an important priority in emergency departments and other settings. At the same time, many young patients may not disclose risk behaviors or even engage in discussions of HIV testing when they interact with healthcare providers because they may fear being stigmatized. Technology-based interventions present valuable opportunities to reframe risk reporting and discussions of testing by designing computer-mediated interactions that young sexual and racial minority participants find non-judgmental and less threatening. If designed in accordance with empirically tested theories of instructional design/multimedia learning and established models of behavior change, technology-based interventions can increase the number of HIV tests offered to young people and offer testing in nonthreatening ways that more young people will accept. The current paper describes a hyper-iterative methodology used to develop the Mobile Augmented Screening (MAS) tool, a technology-based intervention designed to destigmatize HIV and increase HIV test rates among youth.


Soot in urban atmospheres: determination by an optical absorption technique.
We have used the high optical absorptivity of urban and source particulates to trace their "graphitic" component. The optical absorptivity and the particulate carbon loading show a strong correlation. Analyses of the data indicate that primary soot emissions compose a major fraction of the carbonaceous aerosol and put a low limit on secondary organic material produced in correlation with the ozone concentration.


RNA Recombination Enhances Adaptability and Is Required for Virus Spread and Virulence.
Mutation and recombination are central processes driving microbial evolution. A high mutation rate fuels adaptation but also generates deleterious mutations. Recombination between two different genomes may resolve this paradox, alleviating effects of clonal interference and purging deleterious mutations. Here we demonstrate that recombination significantly accelerates adaptation and evolution during acute virus infection. We identified a poliovirus recombination determinant within the virus polymerase, mutation of which reduces recombination rates without altering replication fidelity. By generating a panel of variants with distinct mutation rates and recombination ability, we demonstrate that recombination is essential to enrich the population in beneficial mutations and purge it from deleterious mutations. The concerted activities of mutation and recombination are key to virus spread and virulence in infected animals. These findings inform a mathematical model to demonstrate that poliovirus adapts most rapidly at an optimal mutation rate determined by the trade-off between selection and accumulation of detrimental mutations.