Evaluation and management of the AGUS Papanicolaou smear.
Atypical glandular cells on Papanicolaou smears are an unusual but important cytologic diagnosis. The Bethesda system classifies atypical glandular cells of undetermined significance (AGUS) as glandular cells that demonstrate nuclear atypia appearing to exceed reactive or reparative changes but lacking unequivocal features of adenocarcinoma. AGUS occurs in approximately 0.18 to 0.74 percent of all cervical smears. Because of the high likelihood that AGUS is associated with significant clinical disease, simply repeating the Papanicolaou smear is not sufficient for the management of AGUS. Unlike women with atypical squamous cells of undetermined significance, a significant percentage of women with AGUS will have more serious lesions, such as high-grade preinvasive squamous disease, adenocarcinoma in situ, adenocarcinoma or invasive cancers from sites other than the cervix. Colposcopic examination is recommended for all women with a cytologic diagnosis of AGUS. Those women with AGUS that is suspicious for adenocarcinoma should undergo cervical conization, even in the absence of detectable abnormalities on colposcopic examination.


Using Smartphones and Machine Learning to Quantify Parkinson Disease Severity: The Mobile Parkinson Disease Score.
Current Parkinson disease (PD) measures are subjective, rater-dependent, and assessed in clinic. Smartphones can measure PD features, yet no smartphone-derived rating score exists to assess motor symptom severity in real-world settings. To develop an objective measure of PD severity and test construct validity by evaluating the ability of the measure to capture intraday symptom fluctuations, correlate with current standard PD outcome measures, and respond to dopaminergic therapy. This observational study assessed individuals with PD who remotely completed 5 tasks (voice, finger tapping, gait, balance, and reaction time) on the smartphone application. We used a novel machine-learning-based approach to generate a mobile Parkinson disease score (mPDS) that objectively weighs features derived from each smartphone activity (eg, stride length from the gait activity) and is scaled from 0 to 100 (where higher scores indicate greater severity). Individuals with and without PD additionally completed standard in-person assessments of PD with smartphone assessments during a period of 6 months. Ability of the mPDS to detect intraday symptom fluctuations, the correlation between the mPDS and standard measures, and the ability of the mPDS to respond to dopaminergic medication. The mPDS was derived from 6148 smartphone activity assessments from 129 individuals (mean [SD] age, 58.7 [8.6] years; 56 [43.4%] women). Gait features contributed most to the total mPDS (33.4%). In addition, 23 individuals with PD (mean [SD] age, 64.6 [11.5] years; 11 [48%] women) and 17 without PD (mean [SD] age 54.2 [16.5] years; 12 [71%] women) completed in-clinic assessments. The mPDS detected symptom fluctuations with a mean (SD) intraday change of 13.9 (10.3) points on a scale of 0 to 100. The measure correlated well with the Movement Disorder Society Unified Parkinson Disease's Rating Scale total (r = 0.81; P < .001) and part III only (r = 0.88; P < .001), the Timed Up and Go assessment (r = 0.72; P = .002), and the Hoehn and Yahr stage (r = 0.91; P < .001). The mPDS improved by a mean (SD) of 16.3 (5.6) points in response to dopaminergic therapy. Using a novel machine-learning approach, we created and demonstrated construct validity of an objective PD severity score derived from smartphone assessments. This score complements standard PD measures by providing frequent, objective, real-world assessments that could enhance clinical care and evaluation of novel therapeutics.


The tumor suppressor Ikaros shapes the repertoire of notch target genes in T cells.
The Notch signaling pathway is activated in many cell types, but its effects are cell type- and stage-specific. In the immune system, Notch activity is required for the differentiation of T cell progenitors, but it is reduced in more mature thymocytes, in which Notch is oncogenic. Studies based on single-gene models have suggested that the tumor suppressor protein Ikaros plays an important role in repressing the transcription of Notch target genes. We used genome-wide analyses, including chromatin immunoprecipitation sequencing, to identify genes controlled by Notch and Ikaros in gain- and loss-of-function experiments. We found that Ikaros bound to and directly repressed the expression of most genes that are activated by Notch. Specific deletion of Ikaros in thymocytes led to the persistent expression of Notch target genes that are essential for T cell maturation, as well as the rapid development of T cell leukemias in mice. Expression of Notch target genes that are normally silent in T cells, but are activated by Notch in other cell types, occurred in T cells of mice genetically deficient in Ikaros. We propose that Ikaros shapes the timing and repertoire of the Notch transcriptional response in T cells through widespread targeting of elements adjacent to Notch regulatory sequences. These results provide a molecular framework for understanding the regulation of tissue-specific and tumor-related Notch responses.


Development of Monoclonal Antibodies against CMP-N-Acetylneuraminate-beta-galactosamide-alpha-2,3-sialyltransferase 1 (ST3Gal-I) Recombinant Protein Expressed in E. coli.
Aberrant glycosylation is one of the major hallmarks of cancer with altered gene expression signatures of sialyltransferases. ST3Gal-I, a sialyltransferase, is known to play a crucial role in sialylation of T antigen in bladder cancer and it has reported elevated expression in breast carcinogenesis with increased tumor progression stages. The aim of the current study is to develop new monoclonal antibodies (mAbs) against human ST3Gal-I and evaluate their diagnostic potential. We developed a repertoire of stable hybridoma cell lines producing high-affinity IgG antibodies against recombinant human ST3Gal-I, expressed in E. coli BL21-DE3 strain. In order to demonstrate the diagnostic value of the mAbs, various clones were employed for the immunohistochemistry analysis of ST3Gal-I expression in cancerous tissues. Antibodies generated by 7E51C83A10 clone demonstrated a strong and specific fluorescence staining in breast cancer tissue sections and did not exhibit significant background in fibroadenoma sections. In conclusion, the mAbs raised against recombinant ST3Gal-I recognize cellular ST3Gal-I and represent a promising diagnostic tool for the immunodetection of ST3Gal-I expressing cells. Specific-reactivity of clone 7E51C83A10 mAbs towards ST3Gal-I was also confirmed by immunoblotting. Therefore, our observations warrant evaluation of ST3Gal-I as a potential marker for cancer diagnosis at larger scale.


Hospice Admission and Survival After 18F-Fluoride PET Performed for Evaluation of Osseous Metastatic Disease in the National Oncologic PET Registry.
We have previously reported that PET using 18F-fluoride (NaF PET) for assessment of osseous metastatic disease was associated with substantial changes in intended management in Medicare beneficiaries participating in the National Oncologic PET Registry (NOPR). Here, we use Medicare administrative data to examine the association between NaF PET results and hospice claims within 180 d and 1-y survival. Methods: We classified NOPR NaF PET results linked to Medicare claims by imaging indication (initial staging [IS]; detection of suspected first osseous metastasis [FOM]; suspected progression of osseous metastasis [POM]; or treatment monitoring [TM]) and type of cancer (prostate, lung, breast, or other). Results were classified as definitely positive scan findings versus probably positive scan findings versus negative scan findings for osseous metastasis for IS and FOM; more extensive disease versus no change or less extensive disease for POM; and worse prognosis versus no change or better prognosis for TM, based on the postscan assessment. Our study included 21,167 scans obtained from 2011 to 2014 of consenting NOPR participants aged 65 y or older. Results: The relative risk of hospice claims within 180 d of a NaF PET scan was 2.0-7.5 times higher for patients with evidence of new or progressing osseous metastasis than for those without, depending on indication and cancer type (all P < 0.008). The percentage difference in hospice claims for those with a finding of new or more advanced osseous disease ranged from 3.9% for IS prostate patients to 28% for FOM lung patients. Six-month survival was also associated with evidence of new or increased osseous disease; risk of death was 1.8-5.1 times as likely (all P ≤ 0.0001), with percentage differences of approximately 30% comparing positive and negative scans in patients with lung cancer imaged for IS or FOM. Conclusion: Our analyses demonstrated that NaF PET scan results are highly associated with subsequent hospice claims and, ultimately, with patient survival. NaF PET provides important information on the presence of osseous metastasis and prognosis to assist patients and their physicians when making decisions on whether to select palliative care and transition to hospice or whether to continue treatment.


Absolute Improvements in Freedom From Distant Recurrence to Tailor Adjuvant Endocrine Therapies for Premenopausal Women: Results From TEXT and SOFT.
The Tamoxifen and Exemestane Trial (TEXT)/Suppression of Ovarian Function Trial (SOFT) showed superior outcomes for premenopausal women with hormone receptor (HR)-positive breast cancer treated with adjuvant exemestane plus ovarian function suppression (OFS) or tamoxifen plus OFS versus tamoxifen alone. We previously reported the magnitude of absolute improvements in freedom from any recurrence across a continuous, composite measure of recurrence risk to tailor decision making. With longer follow-up, we now focus on distant recurrence. The TEXT/SOFT HR-positive/human epidermal growth factor receptor 2 (HER2)-negative analysis population included 4,891 women stratified by predetermined chemotherapy use. Kaplan-Meier estimates of 8-year freedom from distant recurrence were analyzed using subpopulation treatment effect pattern plot (STEPP) methodology across subpopulations defined by the continuous composite measure of recurrence risk. For each patient, the composite risk value was obtained from a Cox model that incorporated age; nodal status; tumor size; grade; and estrogen receptor, progesterone receptor, and Ki-67 labeling index expression levels. The overall rate of 8-year freedom from distant recurrence was 91.1% and ranged from approximately 100% to 63% across lowest to highest composite risks. TEXT patients who received chemotherapy had an average absolute improvement with exemestane plus OFS versus tamoxifen plus OFS of 5.1%, and STEPP analysis showed improvements from less than 1% to more than 15% from lowest to highest composite risks. SOFT patients who remained premenopausal after chemotherapy had an average 5.2% absolute improvement with exemestane plus OFS versus tamoxifen and reached 10% across composite risks; for tamoxifen plus OFS versus tamoxifen, the maximum improvement was approximately 3.5%. Women who did not receive chemotherapy had a more than 97% rate of 8-year freedom from distant recurrence, and improvements with exemestane plus OFS ranged from 1% to 4%. Premenopausal women with HR-positive/HER2-negative breast cancer and high recurrence risk, as defined by clinicopathologic characteristics, may experience a 10% to 15% absolute improvement in 8-year freedom from distant recurrence with exemestane plus OFS versus tamoxifen plus OFS or tamoxifen alone. The potential benefit of escalating endocrine therapy versus tamoxifen alone is minimal for those at low recurrence risk.


Systematic variation of length ratio and the formation of smectic A and smectic C phases.
The phase diagrams of four binary mixtures of chemically similar smectogenic mesogens differing only in molecular length are investigated. In these bidisperse systems the length ratio varies systematically. The phase diagrams show the stabilization of the smectic A and the destabilization of the smectic C phase with increasing length ratio as a general trend. Detailed small-angle X-ray diffraction and electro-optic measurements revealed a decrease in smectic translational order and a continuous reduction of the tilt angle with increasing length difference. These surprising results are of general interest for the understanding of the structure and dynamics of smectic phases. The remarkably strong impact of the length difference on the smectic layer structure and the phase behavior is discussed from a mechanistic point of view taking into account sterical interactions. For the observed structural changes in these bidisperse smectics we propose pronounced out-of-layer fluctuations with increasing length difference as driving force, causing neighboring molecules within nearest layer into a smectic A-like packing.


Alcohol-and-HIV-Induced Lysosomal Dysfunction Regulates Extracellular Vesicles Secretion in Vitro and in Liver-Humanized Mice.
Alcohol abuse is common in people living with HIV-1 and dramaticallyenhances the severity of HIV-induced liver damage by inducing oxidative stress and lysosomaldysfunction in the liver cells. We hypothesize that the increased release of extracellular vesicles(EVs) in hepatocytes and liver humanized mouse model is linked to lysosome dysfunction. The study was performed on primary human hepatocytes and human hepatoma RLWXP-GFP (Huh7.5 cells stably transfected with CYP2E1 and XPack-GFP) cells and validated on ethanol-fed liverhumanizedfumarylacetoacetate hydrolase (Fah)-/-, Rag2-/-, common cytokine receptor gamma chainknockout (FRG-KO) mice. Cells and mice were infected with HIV-1ADA virus. We observedan increase in the secretion of EVs associated with a decrease in lysosomal activity and expressionof lysosomal-associated membrane protein 1. Next-generation RNA sequencing of primary humanhepatocytes revealed 63 differentially expressed genes, with 13 downregulated and 50 upregulatedgenes in the alcohol-HIV-treated group. Upstream regulator analysis of differentially expressedgenes through Ingenuity Pathway Analysis identified transcriptional regulators affecting downstreamgenes associated with increased oxidative stress, lysosomal associated disease, and function andEVs biogenesis. Our in vitro findings were corroborated by in vivo studies on human hepatocytetransplantedhumanized mice, indicating that intensive EVs' generation by human hepatocytes andtheir secretion to serum was associated with increased oxidative stress and reduction in lysosomalactivities triggered by HIV infection and ethanol diet. HIV-and-ethanol-metabolisminducedEVs release is tightly controlled by lysosome status in hepatocytes and participates in thedevelopment of double-insult-induced liver injury.


Mesenchymal Stem Cells Shift Mitochondrial Dynamics and Enhance Oxidative Phosphorylation in Recipient Cells.
Mesenchymal stem cells (MSCs) are the most commonly used cells in tissue engineering and regenerative medicine. MSCs can promote host tissue repair through several different mechanisms including donor cell engraftment, release of cell signaling factors, and the transfer of healthy organelles to the host. In the present study, we examine the specific impacts of MSCs on mitochondrial morphology and function in host tissues. Employing in vitro cell culture of inherited mitochondrial disease and an in vivo animal experimental model of low-grade inflammation (high fat feeding), we show human-derived MSCs to alter mitochondrial function. MSC co-culture with skin fibroblasts from mitochondrial disease patients rescued aberrant mitochondrial morphology from a fission state to a more fused appearance indicating an effect of MSC co-culture on host cell mitochondrial network formation. In vivo experiments confirmed mitochondrial abundance and mitochondrial oxygen consumption rates were elevated in host tissues following MSC treatment. Furthermore, microarray profiling identified 226 genes with differential expression in the liver of animals treated with MSC, with cellular signaling, and actin cytoskeleton regulation as key upregulated processes. Collectively, our data indicate that MSC therapy rescues impaired mitochondrial morphology, enhances host metabolic capacity, and induces widespread host gene shifting. These results highlight the potential of MSCs to modulate mitochondria in both inherited and pathological disease states.


Production of IFN-γ in feline whole blood after incubation with potential T-cell epitopes of the nucleocapsid protein of feline coronavirus.
Interferon gamma (IFN-γ) plays an important role in cell mediated responses against mutated feline coronavirus strains (FCoV) involved in the pathogenesis of feline infectious peritonitis (FIP). The aim of this study was to establish a combined in silico and in vitro approach to assess feline leukocyte production of IFN-γ in response to selected peptides of the nucleocapside protein (N) of FCoVs. To this aim, we designed, through a bioinformatic approach, 8 potentially immunogenic peptides from the protein N corresponding to sequences of residues 14, 182, 198 detected only in FCoVs from FIP cats (virulent strains), only in FCoVs from healthy cats (avirulent strains) and both in FIP and in healthy cats (mixed strains). The peptides or a sham solution were incubated with whole blood from 16 cats (7 healthy and 9 with chronic diseases other than FIP) and IFN-γ concentration was measured on plasma using an ELISA system. RT-PCR expression of IFN-γ mRNA was also evaluated after incubation of the peptides or a sham solution with whole blood from 4 clinically healthy cats. The mean plasma concentration of IFN-γ in samples incubated with peptides decreased and the expression of IFN-γmRNA did not change compared with the sham solution, except for some cats with chronic diseases (which probably have a "pre-activated" immune response). These cats responded to "avirulent" or "mixed" peptides by increasing the concentration of IFN-γ and the expression of IFN-γ mRNA. The combined approach employed in this study allowed us to identify potentially immunogenic peptides of FCoV N protein that can modulate the production of IFN-γ especially in cats with a "pre-activated" cell mediated response.


Human CD8+ T cell cross-reactivity across influenza A, B and C viruses.
Influenza A, B and C viruses (IAV, IBV and ICV, respectively) circulate globally and infect humans, with IAV and IBV causing the most severe disease. CD8+ T cells confer cross-protection against IAV strains, however the responses of CD8+ T cells to IBV and ICV are understudied. We investigated the breadth of CD8+ T cell cross-recognition and provide evidence of CD8+ T cell cross-reactivity across IAV, IBV and ICV. We identified immunodominant CD8+ T cell epitopes from IBVs that were protective in mice and found memory CD8+ T cells directed against universal and influenza-virus-type-specific epitopes in the blood and lungs of healthy humans. Lung-derived CD8+ T cells displayed tissue-resident memory phenotypes. Notably, CD38+Ki67+CD8+ effector T cells directed against novel epitopes were readily detected in IAV- or IBV-infected pediatric and adult subjects. Our study introduces a new paradigm whereby CD8+ T cells confer unprecedented cross-reactivity across all influenza viruses, a key finding for the design of universal vaccines.


Cholecalciferol supplementation alters calcitriol-responsive monocyte proteins and decreases inflammatory cytokines in ESRD.
In vitro, monocyte 1alpha-hydroxylase converts 25-hydroxyvitamin D [25(OH)D] to 1,25-dihydroxyvitamin D to regulate local innate immune responses, but whether 25(OH)D repletion affects vitamin D-responsive monocyte pathways in vivo is unknown. Here, we identified seven patients who had 25(OH)D insufficiency and were undergoing long-term hemodialysis and assessed changes after cholecalciferol and paricalcitol therapies in both vitamin D-responsive proteins in circulating monocytes and serum levels of inflammatory cytokines. Cholecalciferol therapy increased serum 25(OH)D levels four-fold, monocyte vitamin D receptor expression three-fold, and 24-hydroxylase expression; therapy decreased monocyte 1alpha-hydroxylase levels. The CD16(+) "inflammatory" monocyte subset responded to 25(OH)D repletion the most, demonstrating the greatest increase in vitamin D receptor expression after cholecalciferol. Cholecalciferol therapy reduced circulating levels of inflammatory cytokines, including IL-8, IL-6, and TNF. These data suggest that nutritional vitamin D therapy has a biologic effect on circulating monocytes and associated inflammatory markers in patients with ESRD.


Environmental Pollutant Ozone Causes Damage to Lung Surfactant Protein B (SP-B).
Lung surfactant protein B (SP-B) is an essential protein found in the surfactant fluid at the air-water interface of the lung. Exposure to the air pollutant ozone could potentially damage SP-B and lead to respiratory distress. We have studied two peptides, one consisting of the N-terminus of SP-B [SP-B(1-25)] and the other a construct of the N- and C-termini of SP-B [SP-B(1-25,63-78)], called SMB. Exposure to dilute levels of ozone (~2 ppm) of monolayers of each peptide at the air-water interface leads to a rapid reaction, which is evident from an increase in the surface tension. Fluorescence experiments revealed that this increase in surface tension is accompanied by a loss of fluorescence from the tryptophan residue at the interface. Neutron and X-ray reflectivity experiments show that, in contrast to suggestions in the literature, the peptides are not solubilized upon oxidation but rather remain at the interface with little change in their hydration. Analysis of the product material reveals that no cleavage of the peptides occurs, but a more hydrophobic product is slowly formed together with an increased level of oligomerization. We attributed this to partial unfolding of the peptides. Experiments conducted in the presence of phospholipids reveal that the presence of the lipids does not prevent oxidation of the peptides. Our results strongly suggest that exposure to low levels of ozone gas will damage SP-B, leading to a change in its structure. The implication is that the oxidized protein will be impaired in its ability to interact at the air-water interface with negatively charged phosphoglycerol lipids, thus compromising what is thought to be its main biological function.


Extending the CARE Principles from tribal research policies to benefit sharing in genomic research.
Indigenous Peoples have historically been targets of extractive research that has led to little to no benefit. In genomics, such research not only exposes communities to harms and risks of misuse, but also deprives such communities of potential benefits. Tribes in the US have been exercising their sovereignty to limit this extractive practice by adopting laws and policies to govern research on their territories and with their citizens. Federally and state recognized tribes are in the strongest position to assert research oversight. Other tribes lack the same authority, given that federal and state governments do not recognize their rights to regulate research, resulting in varying levels of oversight by tribes. These governance measures establish collective protections absent from the US federal government's research oversight infrastructure, while setting expectations regarding benefits to tribes as political collectives. Using a legal epidemiology approach, the paper discusses findings from a review of Tribal research legislation, policy, and administrative materials from 26 tribes in the US. The discussion specifies issues viewed by tribes as facilitators and barriers to securing benefits from research for their nations and members/citizens, and describes preemptive and mitigating strategies pursued by tribes in response. These strategies are set within the framing of the CARE Principles for Indigenous Data Governance (Collective Benefit, Authority to Control, Responsibility, Ethics), a set of standards developed to ensure that decisions made about data pertaining to Indigenous communities at the individual and tribal levels are responsive to their values and collective interests. Our findings illustrate gaps to address for benefit sharing and a need to strengthen Responsibility and Ethics in tribal research governance.


Declining trends of PCDD/Fs in lichens over a decade in a Mediterranean area with multiple pollution sources.
Lichens are one of the most useful environmental biomonitors, due to their ability to clearly reflect atmospheric deposition of pollutants. Dioxin and furan (PCDD/F) emissions have been reported to be decreasing in North European countries as a consequence of European regulations. This reduction has been perceptible across several environmental matrices, but it hasn't yet been shown in lichens as typical biomonitors of atmospheric pollution. In this work we compared concentrations of PCDD/Fs in two lichen species collected in a Mediterranean area with mixed land-uses, encompassing urban, industrial and natural areas, in 2009 and 2011 with the ones obtained in the same species collected in the same region in 2000. We found that PCDD/F concentrations in both lichen species have decreased approximately 70% since 2000 whereas industrial emissions have only decreased 25% for the same period. This substantial greater reduction observed in lichens may be due to several causes; after excluding fires as a possible explanation, we point out that possible causes could not only be the overall decrease in industrial emissions but also other causes such as traffic reduction and/or increase efficiency in the use of fuels. Capsule: PCDD/F concentrations in lichens have decreased 70% over the last decade, whereas industrial emissions have only decreased 25%.


Conserved enhancer and silencer elements responsible for differential Adh transcription in Drosophila cell lines.
The distal promoter of Adh is differentially expressed in Drosophila tissue culture cell lines. After transfection with an exogenous Adh gene, there was a specific increase in distal alcohol dehydrogenase (ADH) transcripts in ADH-expressing (ADH+) cells above the levels observed in transfected ADH-nonexpressing (ADH-) cells. We used deletion mutations and a comparative transient-expression assay to identify the cis-acting elements responsible for enhanced Adh distal transcription in ADH+ cells. DNA sequences controlling high levels of distal transcription were localized to a 15-base-pair (bp) region nearly 500 bp upstream of the distal RNA start site. In addition, a 61-bp negative cis-acting element was found upstream from and adjacent to the enhancer. When this silencer element was deleted, distal transcription increased only in the ADH+ cell line. These distant upstream elements must interact with the promoter elements, the Adf-1-binding site and the TATA box, as they only influenced transcription when at least one of these two positive distal promoter elements was present. Internal deletions targeted to the Adf-1-binding site or the TATA box reduced transcription in both cell types but did not affect the transcription initiation site. Distal transcription in transfected ADH- cells appears to be controlled primarily through these promoter elements and does not involve the upstream regulatory elements. Evolutionary conservation in distantly related Drosophila species suggests the importance of these upstream elements in correct developmental and tissue-specific expression of ADH.


Selenoprotein S regulates adipogenesis through IRE1α-XBP1 pathway.
The induction of endoplasmic reticulum (ER) stress is associated with adipogenesis, during which the inositol-requiring enzyme 1 alpha (IRE1α)-X-box-binding protein 1 (XBP1) pathway is involved. Selenoprotein S (SelS), which is an ER resident selenoprotein, is involved in ER homeostasis regulation; however, little is known about the role of SelS in regulating adipogenesis. In vivo studies showed that SelS protein levels in white adipose tissue were increased in obese subjects and high-fat diet (HFD)-fed mice. Moreover, we identified that SelS protein levels increased in the early phase of adipogenesis and then decreased in the late phase during adipogenesis. Overexpression of SelS promoted adipogenesis. Conversely, knockdown (KD) of SelS resulted in the inhibition of adipogenesis, which was related to increasing cell death, decreased mitotic clonal expansion, and cell cycle G1 arrest. In vivo studies also showed that ER stress markers (p-IRE1α/IRE1α, XBP1s, and Grp78) were significantly increased with upregulating of SelS expression in subcutaneous and visceral adipose tissues in the obese subjects and HFD-fed mice. Furthermore, in SelS KD cells, the levels of Grp78 were increased and the levels of p-IRE1α/IRE1α were unchanged , but mRNA levels of spliced XBP1 (XBP1s) produced by IRE1α-mediated splicing were decreased, suggesting a role of SelS in the modulation of IRE1α-XBP1 pathway. Moreover, inhibition of adipogenesis by SelS suppression can be rescued by overexpression of XBP1s. Thus, SelS appears to function as a novel regulator of adipogenesis through the IRE1α-XBP1 signaling pathway.


An apodized SOI waveguide-to-fiber surface grating coupler for single lithography silicon photonics.
We present the design, fabrication, and characterization of a grating for coupling between a single mode silica fiber and the TE mode in a silicon photonic waveguide on a silicon on insulator (SOI) substrate. The grating is etched completely through the silicon device layer, thus permitting the fabrication of through-etched surface coupled silicon nanophotonic circuits in a single lithography step. Furthermore, the grating is apodized to match the diffracted wave to the mode profile of the fiber. We experimentally demonstrate a coupling efficiency of 35% with a 1 dB bandwidth of 47 nm at 1536 nm on a standard SOI substrate. Furthermore, we show by simulation that with an optimized buried oxide thickness, a coupling efficiency of 72% and a 1 dB bandwidth of 38 nm at 1550 nm is achievable. This is, to our knowledge, the highest simulated coupling efficiency for single-etch TE-mode grating couplers. In particular, simulations show that apodizing a conventional periodic through-etched grating decreases the back-reflection into the waveguide from 21% to 0.1%.


Insights into the deviation from piecewise linearity in transition metal complexes from supervised machine learning models.
Virtual high-throughput screening (VHTS) and machine learning (ML) with density functional theory (DFT) suffer from inaccuracies from the underlying density functional approximation (DFA). Many of these inaccuracies can be traced to the lack of derivative discontinuity that leads to a curvature in the energy with electron addition or removal. Over a dataset of nearly one thousand transition metal complexes typical of VHTS applications, we computed and analyzed the average curvature (i.e., deviation from piecewise linearity) for 23 density functional approximations spanning multiple rungs of "Jacob's ladder". While we observe the expected dependence of the curvatures on Hartree-Fock exchange, we note limited correlation of curvature values between different rungs of "Jacob's ladder". We train ML models (i.e., artificial neural networks or ANNs) to predict the curvature and the associated frontier orbital energies for each of these 23 functionals and then interpret differences in curvature among the different DFAs through analysis of the ML models. Notably, we observe spin to play a much more important role in determining the curvature of range-separated and double hybrids in comparison to semi-local functionals, explaining why curvature values are weakly correlated between these and other families of functionals. Over a space of 187.2k hypothetical compounds, we use our ANNs to pinpoint DFAs for which representative transition metal complexes have near-zero curvature with low uncertainty, demonstrating an approach to accelerate screening of complexes with targeted optical gaps.


Whole-genome resequencing of extreme phenotypes in collared flycatchers highlights the difficulty of detecting quantitative trait loci in natural populations.
Dissecting the genetic basis of phenotypic variation in natural populations is a long-standing goal in evolutionary biology. One open question is whether quantitative traits are determined only by large numbers of genes with small effects, or whether variation also exists in large-effect loci. We conducted genomewide association analyses of forehead patch size (a sexually selected trait) on 81 whole-genome-resequenced male collared flycatchers with extreme phenotypes, and on 415 males sampled independent of patch size and genotyped with a 50K SNP chip. No SNPs were genomewide statistically significantly associated with patch size. Simulation-based power analyses suggest that the power to detect large-effect loci responsible for 10% of phenotypic variance was <0.5 in the genome resequencing analysis, and <0.1 in the SNP chip analysis. Reducing the recombination by two-thirds relative to collared flycatchers modestly increased power. Tripling sample size increased power to >0.8 for resequencing of extreme phenotypes (N = 243), but power remained <0.2 for the 50K SNP chip analysis (N = 1245). At least 1 million SNPs were necessary to achieve power >0.8 when analysing 415 randomly sampled phenotypes. However, power of the 50K SNP chip to detect large-effect loci was nearly 0.8 in simulations with a small effective population size of 1500. These results suggest that reliably detecting large-effect trait loci in large natural populations will often require thousands of individuals and near complete sampling of the genome. Encouragingly, far fewer individuals and loci will often be sufficient to reliably detect large-effect loci in small populations with widespread strong linkage disequilibrium.


A qualitative systematic review of experiences and perceptions of youth suicide.
Suicide remains a global issue with over 800,000 people dying from suicide every year. Youth suicide is especially serious due to the years of life lost when a young person takes their own life. Social interactions, perceived support, genetic predisposition and mental illnesses are factors associated with suicide ideation. To review and synthesize qualitative studies that explored the experiences and perceptions of suicide in people 25 years old and younger. Qualitative systematic review. PubMed, PsycINFO, Scopus and CINAHL were searched alongside hand-searching reference lists up to October 2018. Methodological quality was assessed using the qualitative Critical Appraisal Skills Programme checklist. The 27 studies included in the review centered around youth suicide and included interviews with young people and members of the wider community. Thematic synthesis focused on factors leading to suicide attempts, elements important to recovery, beliefs within the community, and treatment/prevention strategies. Thematic analysis of the articles revealed four categories: i) triggers and risks leading to suicidality; ii) factors involved in recovery; iii) need for institutional treatment/prevention strategies; and iv) beliefs about suicide at a community level. The first category was further subdivided into: i) behaviours; ii) feelings/emotions; iii) family influences; iv) peer influences; and v) other. The second category was split into: i) interpersonal; ii) cultural; and iii) individual influences, while the third category was divided into i) education; and ii) treatment. Youth suicide is a complex issue with many causes and risks factors which interact with one another. For successful treatment and prevention, procedural reform is needed, along with a shift in societal attitudes toward emotional expression and suicide.


Functional balance in older adults with chronic obstructive pulmonary disease.
This study assessed functional balance among older adults at all grades of chronic obstructive pulmonary disease (COPD) and explored balance impairment predictors. A cross-sectional study with outpatients with COPD (N = 160; M = 72.2 years, SD = 7.9; mean forced expiratory volume in 1 s = 63.8% predicted, SD = 23.7) was conducted. The Timed Up and Go (TUG) test was used to assess functional balance. Functional balance impairment was defined as a TUG score exceeding the upper limit of the confidence intervals of normative values for healthy older adults. Participants performed the TUG test in 11.0 s (SD = 4.8 s). Functional balance impairment was present in 44.4% of the participants and was significantly more frequent in severe to very severe COPD (62.5%). Body mass index (odds ratio [OR] = 1.12), number of medications (OR = 1.20), restriction in recreational activities (OR = 1.66), and depression score (OR = 1.14) were multivariate predictors of functional balance impairment. Functional balance impairment is present in early COPD, although more evident at advanced grades. These findings highlight the importance of balance assessment in older patients at all COPD grades.


Night time resistance exercise alters muscular IL-6-related protein signaling, but not muscle growth after 10 weeks of resistance training in male rats.
The present study investigated the time-of-day effects on acute response and chronic adaptations to resistance exercise (RE) in rat skeletal muscle. Male rats were divided into Early and Late training groups and performed climbing RE during the first or last hour of the active (dark) period, respectively. The first experiment measured muscle mass and strength after a 10-week climbing training program. The second experiment examined inflammatory signaling response and satellite cell (SC) numbers following an acute bout of RE. The results showed no significant differences between rats training at early and late active periods in relative muscle weight (muscle-to-weight ratio), cross sectional area (CSA) and strength. The acute study observed increased STAT1 phosphorylation, oxidative stress (2-thiobarbituric acid reacting substances, TBARS), SCs (Pax7+), neutrophils (His48+) and macrophages (CD68+), and decreased interleukin 6 (IL-6) protein expression of skeletal muscle relative to non-exercise control after an acute bout of RE. Interestingly, higher plasma IL-6 and STAT3 phosphorylation response was observed in the late training group when compared to the early training group after an acute bout of RE. The results of this study suggest that animals can adapt to resistance training at different time-of-day, by modulating inflammatory signaling of skeletal muscle.


Contrast Sensitivity After Wavefront-Guided and Wavefront-Optimized PRK and LASIK for Myopia and Myopic Astigmatism.
To compare contrast sensitivity among participants undergoing wavefront-guided or wavefront-optimized photorefractive keratectomy (PRK) or LASIK for the treatment of myopia or myopic astigmatism 12 months after surgery. In a prospective, randomized clinical trial, 215 participants with myopia ranging from -0.50 to -7.25 diopters (D) and less than -3.50 D of manifest astigmatism electing to undergo either LASIK or PRK were randomized to receive wavefront-guided or wavefront-optimized treatment. Corrected Super Vision Test (Precision Vision, La Salle, IL) high contrast and small letter contrast sensitivity, uncorrected postoperative contrast sensitivity function, and uncorrected and corrected distance visual acuity were measured preoperatively and at 1, 3, 6, and 12 months postoperatively. There was a significant difference within each of the four groups over time when measuring high contrast visual acuity (P < .001) and small letter contrast sensitivity (P < .001), with the most significant decrease occurring 1 month postoperatively. However, there were no significant differences when comparing the four groups for high contrast sensitivity (P = .22) or small letter contrast sensitivity (P = .06). The area under the logarithm of contrast sensitivity function did not differ significantly over time (P = .09) or between groups (P = .16). A pairwise comparison of preoperative to 12-month CDVA showed a significant improvement in all groups (P < .017). The change in CDVA was also significantly different between groups as determined by one-way analysis of variance (P = .003). Wavefront-guided and wavefront-optimized PRK and LASIK procedures maintained high contrast, small letter contrast sensitivity, and contrast sensitivity function 12 months postoperatively. Although the recovery period for visual performance was longer for PRK versus LASIK, there was no significant difference in treatment type or treatment profile at 12 months postoperatively. [J Refract Surg. 2018;34(9):590-596.].


Influence of personal characteristics of individual women on sensitivity and specificity of mammography in the Million Women Study: cohort study.
To examine how lifestyle, hormonal, and other factors influence the sensitivity and specificity of mammography. Women recruited into the Million Women Study completed a questionnaire about various personal factors before routine mammographic screening. A sample of 122,355 women aged 50-64 years were followed for outcome of screening and incident breast cancer in the next 12 months. Sensitivity and specificity were calculated by using standard definitions, with adjustment for potential confounding factors. Breast cancer was diagnosed in 726 (0.6%) women, 629 in screen positive and 97 in screen negative women; 3885 (3.2%) were screen positive but had no subsequent diagnosis of breast cancer. Overall sensitivity was 86.6% and specificity was 96.8%. Three factors had an adverse effect on both measures: use of hormone replacement therapy (sensitivity: 83.0% (95% confidence interval 77.4% to 87.6%), 84.7% (73.9% to 91.6%), and 92.1% (87.6% to 95.0%); specificity: 96.8% (96.6% to 97.0%), 97.8% (97.5% to 98.0%), and 98.1% (98.0% to 98.2%), respectively, for current, past, and never use); previous breast surgery v no previous breast surgery (sensitivity: 83.5% (75.7% to 89.1%) v 89.4% (86.5% to 91.8%); specificity: 96.2% (95.8% to 96.5%) v 97.4% (97.3% to 97.5%), respectively); and body mass index < 25 v > or = 25 (sensitivity: 85.7% (81.2% to 89.3%) v 91.0% (87.5% to 93.6%); specificity: 97.2% (97.0% to 97.3%) v 97.4% (97.3% to 97.6%), respectively). Neither sensitivity nor specificity varied significantly according to age, family history of breast cancer, parity, past oral contraceptive use, tubal ligation, physical activity, smoking, or alcohol consumption. The efficiency, and possibly the effectiveness, of mammographic screening is lower in users of hormone replacement therapy, in women with previous breast surgery, and in thin women compared with other women.


Identification of a B-cell surface structure involved in antigen-dependent triggering: absence of this structure on B cells from CBA/N mutant mice.
CBA/N mice have an X-linked B-cell maturation defect which is reflected in part in an absence or dysfunction of a subclass of mature B cells. We have immunized the defective male offspring of the mating (CBA/N female X BALB/c male) with BALB/c spleen cells. The resulting antiserum (alphaLyb3) selectively reacts with a component on the surface of a portion of B cells from a panel of H-2 different mouse strains. Binding of alphaLyb3 serum to this B-cell subclass results in substantial (10- to 20-fold) enhancement of the antibody response to low doses of SRBC. Both binding and enhancing activity are removed by absorption with B cells from B6 and BALB/c, but not CBA/N mice. Absorption of the serum with bone marrow cells, T cells, or thymocytes from Lyb3+ strains does not remove activity. Since the enhanced plaque-forming cell (PFC) responses are specific for the immunizing antigen, and since no PFC response is produced by injection of the antiserum alone, this enhancement probably reflects a second signal produced by specific interaction between antibody and the surface Lyb3 component. Moreover, this signal can partially replace the requirement for T cells in the production of antibody to a "thymus-dependent" antigen. These findings (taken in conjunction with the previously described immune defects in CBA/N mice and other studies of B-cell maturation) suggest to us that Lyb3 is a cell surface component expressed selectively on a mature B-cell subclass. This component is important in B-cell triggering by antigen and fails to develop in CBA/N mice, due to a dysfunction of a regulatory gene on the CBA/N X chromosome.


The validity and reliability of a customized rigid supportive harness during Smith machine back squat exercise.
Although the back squat exercise is commonly prescribed to both athletic and clinical populations, individuals with restricted glenohumeral mobility may be unable to safely support the bar on the upper trapezius using their hands. The aims of this study were to investigate the validity and reliability of a back squat variation using a rigid supportive harness that does not require unrestricted glenohumeral mobility for quantifying 1 repetition maximum (1RM). Thirteen young men (age = 25.3 ± 4.5 years, height = 179.2 ± 6.9 cm, and body mass = 86.6 ± 12.0 kg) with at least 2 years resistance training experience volunteered to participate in the study. Subjects reported to the lab on 3 occasions, each separated by 1 week. During testing sessions, subjects were assessed for 1RM using the traditional back squat (session 1) and harness back squat (HBS; sessions 2 and 3) exercises. Mean 1RM for the traditional back squat, and 2 testing sessions of the HBS (HBS1 and HBS2) were 148.4 ± 25.0 kg, 152.5 ± 25.7 kg, and 150.4 ± 22.6 kg, respectively. Back squat and mean HBS 1RM scores were very strongly correlated (r = 0.96; p < 0.001). There were no significant differences in 1RM scores between the 3 trials. The test-retest 1RM scores with the HBS demonstrated high reliability, with an intraclass correlation coefficient of 0.98 (95% confidence interval [CI] = 0.93-0.99), and a coefficient of variation of 2.6% (95% CI = 1.9-4.3). Taken together, these data suggest that the HBS exercise is a valid and reliable method for assessing 1RM in young men with previous resistance training experience and may be useful for individuals with restricted glenohumeral mobility.


Single-cell metabolomics: changes in the metabolome of freshly isolated and cultured neurons.
Metabolites are involved in a diverse range of intracellular processes, including a cell's response to a changing extracellular environment. Using single-cell capillary electrophoresis coupled to electrospray ionization mass spectrometry, we investigated how placing individual identified neurons in culture affects their metabolic profile. First, glycerol-based cell stabilization was evaluated using metacerebral neurons from Aplysia californica; the measurement error was reduced from ∼24% relative standard deviation to ∼6% for glycerol-stabilized cells compared to those isolated without glycerol stabilization. In order to determine the changes induced by culturing, 14 freshly isolated and 11 overnight-cultured neurons of two metabolically distinct cell types from A. californica, the B1 and B2 buccal neurons, were characterized. Of the more than 300 distinctive cell-related signals detected, 35 compounds were selected for their known biological roles and compared among each measured cell. Unsupervised multivariate and statistical analysis revealed robust metabolic differences between these two identified neuron types. We then compared the changes induced by overnight culturing; metabolite concentrations were distinct for 26 compounds in the cultured B1 cells. In contrast, culturing had less influence on the metabolic profile of the B2 neurons, with only five compounds changing significantly. As a result of these culturing-induced changes, the metabolic composition of the B1 neurons became indistinguishable from the cultured B2 cells. This observation suggests that the two cell types differentially regulate their in vivo or in vitro metabolomes in response to a changing environment.


Suppression of early hematogenous dissemination of human breast cancer cells to bone marrow by retinoic Acid-induced 2.
Regulatory pathways that drive early hematogenous dissemination of tumor cells are insufficiently defined. Here, we used the presence of disseminated tumor cells (DTC) in the bone marrow to define patients with early disseminated breast cancer and identified low retinoic acid-induced 2 (RAI2) expression to be significantly associated with DTC status. Low RAI2 expression was also shown to be an independent poor prognostic factor in 10 different cancer datasets. Depletion of RAI2 protein in luminal breast cancer cell lines resulted in dedifferentiation marked by downregulation of ERα, FOXA1, and GATA3, together with increased invasiveness and activation of AKT signaling. Functional analysis of the previously uncharacterized RAI2 protein revealed molecular interaction with CtBP transcriptional regulators and an overlapping function in controlling the expression of a number of key target genes involved in breast cancer. These results suggest that RAI2 is a new metastasis-associated protein that sustains differentiation of luminal breast epithelial cells. We identified downregulation of RAI2 as a novel metastasis-associated genetic alteration especially associated with early occurring bone metastasis in ERα-positive breast tumors. We specified the role of the RAI2 protein to function as a transcriptional regulator that controls the expression of several key regulators of breast epithelial integrity and cancer.


A review and methodologic critique of the literature refuting whiplash syndrome.
The validity of whiplash syndrome has been a source of debate in the medical literature for many years. Some authors have published articles suggesting that whiplash injuries are impossible at certain collision speeds; others have stated that the problem is psychological, or is feigned as a means to obtain secondary financial gain. These articles contradict the majority of the literature, which shows that whiplash injuries and their sequelae are a highly prevalent problem that affects a significant proportion of the population. The authors of the current literature critique reviewed the biomedical and engineering literature relating to whiplash syndrome, searching for articles that refuted the validity of whiplash injuries. Twenty articles containing nine distinct statements refuting the validity of whiplash syndrome were found that fit the inclusion criteria. The methodology described in these articles was evaluated critically to determine if the authors' observations regarding the validity of whiplash syndrome were scientifically sound. The authors of the current critique found that all of the articles contained significant methodologic flaws with regard to their respective authors' statements refuting the validity of whiplash syndrome. The most frequently found flaws were inadequate study size, nonrepresentative study sample, nonrepresentative crash conditions (for crash tests), and inappropriate study design. As a result of the current literature review, it was determined that there is no epidemiologic or scientific basis in the literature for the following statements: whiplash injuries do not lead to chronic pain, rear impact collisions that do not result in vehicle damage are unlikely to cause injury, and whiplash trauma is biomechanically comparable with common movements of daily living.


Carbon monoxide protects against oxidant-induced apoptosis via inhibition of Kv2.1.
Oxidative stress induces neuronal apoptosis and is implicated in cerebral ischemia, head trauma, and age-related neurodegenerative diseases. An early step in this process is the loss of intracellular K(+) via K(+) channels, and evidence indicates that K(v)2.1 is of particular importance in this regard, being rapidly inserted into the plasma membrane in response to apoptotic stimuli. An additional feature of neuronal oxidative stress is the up-regulation of the inducible enzyme heme oxygenase-1 (HO-1), which catabolizes heme to generate biliverdin, Fe(2+), and carbon monoxide (CO). CO provides neuronal protection against stresses such as stroke and excitotoxicity, although the underlying mechanisms are not yet elucidated. Here, we demonstrate that CO reversibly inhibits K(v)2.1. Channel inhibition by CO involves reactive oxygen species and protein kinase G activity. Overexpression of K(v)2.1 in HEK293 cells increases their vulnerability to oxidant-induced apoptosis, and this is reversed by CO. In hippocampal neurons, CO selectively inhibits K(v)2.1, reverses the dramatic oxidant-induced increase in K(+) current density, and provides marked protection against oxidant-induced apoptosis. Our results provide a novel mechanism to account for the neuroprotective effects of CO against oxidative apoptosis, which has potential for therapeutic exploitation to provide neuronal protection in situations of oxidative stress.


Bilateral disruption of soft tissue extensor mechanism of knee: functional outcome and comparison to unilateral injuries.
Bilateral ruptures of the extensor mechanism are rare. The purpose of this study was to compare the clinical outcomes of operatively treated unilateral and bilateral knee soft tissue extensor mechanism injuries and to identify risk factors for bilateral disruption. All patients operatively treated for a knee extensor mechanism injury were entered into a database and prospectively followed. Postoperative protocol was standardized for all patients. Demographic data, baseline characteristics, range of motion, complications, pain, and functional status were assessed. The main patient-reported outcome measures used in this study were the SF-36 Health Survey and the Lysholm Scale. Patients who sustained bilateral injuries were more likely to have one or more systemic medical conditions. There was no statistical difference between the groups with regard to mechanism of injury or body mass index. The average follow-up was 29 months (range 6-60 months). Patient-reported outcomes, in the form of the SF-36 Health Survey and Lysholm scores, were not significantly different between the two groups at final follow-up. Range of motion and quadriceps strength was also similar between the two cohorts. At latest follow-up, 88% of patients with unilateral injuries and 83% of patients with bilateral disruption were able to return to their pre-injury employment. Operatively treated bilateral knee extensor mechanism disruptions fare similar to unilateral injuries with regard to ultimate functional outcome. The presence of one of more preexisting medical conditions was identified as a risk factor for bilateral tendinous disruption.


Polymorphisms of methylenetetrahydrofolate reductase gene as the genetic predispositions of coronary artery diseases in eastern India.
Gene-environment interaction is an important aspect in the development of coronary artery disease (CAD). The mutation (677C-T) of methylenetetrahydrofolate reductase (MTHFR) gene results in a decrease of the enzyme activity that leads to mild hyperhomocysteinemia. Elevated plasma level of homocysteine has been recognized as an independent risk factor for cardiovascular disease. A case-control study was designed to assess whether the prevalence of some MTHFR gene polymorphisms have any role in the development of CAD. The study included unrelated 217 cases with CAD and 255 healthy controls. DNA was extracted from peripheral blood. MTHFR genotypes were identified by seeing the presence or absence of 677C→T mutation obtained by PCR followed by Hinf1 restriction digestion. Multiple logistic regression analysis was carried out to find association between studied genotypes and lifestyle as well as biochemical risk factors. The T allele was found to be associated with the disease. Significant associations were found with smoking, hypertension, diabetes, and family history of CAD. The results indicate that MTHFR 677C-T polymorphism has significant association with CADs in the population of eastern India.


Expression of FGF and FGF receptor genes in human breast cancer.
The family of FGF growth factors is involved in several biological processes and might play an important role in tumorigenesis. We have studied the respective expression of 8 of the 9 characterized FGF genes, and of the 4 known FGF receptor genes, in a panel of 10 tumor-cell lines and 103 breast-tumor samples, using RT-PCR and Northern-blot analyses. FGF1 and FGF2 were expressed in almost all samples, while expression of FGF5, FGF6, FGF7, and FGF9 was more restricted. FGFR1, FGFR2 and FGFR4 were expressed at high levels in respectively 22%, 4% and 32% of tumors. FGFR3 expression was not detected. The transcript encoding an FGFR1 isoform with 2 immunoglobulin-like domains was the most prevalent.


Effects of shift work on 24-hour ambulatory blood pressure and its variability among Japanese workers.
This study examined the effects of rotating shift work on blood pressure in a comparison of ambulatory blood pressure and long-term changes in blood pressure between shift and day workers. Ambulatory blood pressure was measured for 24-hour periods at an interval of 30 minutes for 27 shift workers and 26 day workers when they worked during the day. Blood pressure was compared between these 2 groups of workers for 4 time categories (awake, sleep, nonwork awake, and work periods). Their long-term blood pressures, recorded in annual surveys, were reviewed for long-term changes. These comparisons were adjusted for the effects of body mass index, alcohol intake, anger expression, and physical activity. On the average, sleep time was shorter and the anger-in (ie, anger suppressed) score was higher for the shift workers than for the day workers, but body mass index and alcohol intake did not differ between the 2 groups. Even after adjustment for these co-variables, the mean systolic blood pressure during the 24-hour, awake, and work periods were higher among the shift workers than among the day workers. The 24-hour standard deviations of the systolic blood pressures were also higher for the shift workers than for the day workers. Among the shift workers, but not among the day workers, a significant long-term increase was observed in systolic blood pressure measured in the annual surveys. These results suggest that shift work may increase systolic blood pressure levels among Japanese men.


Are staffing changes in California's hospitals sensitive to individual hospital characteristics?
Although differences in nurse staffing have been associated with individual hospital characteristics in the literature, there have been no studies on how these factors may influence nurse staffing changes made after the mandated nurse-to-patient ratios in acute care hospitals in California. The aim of this study was to determine if changes in medical-surgical nurse staffing (registered nurses and registry nurses) were associated with particular hospital characteristics. Researchers found the baseline level of nurse staffing was the variable most associated with change in mean productive hours per patient day from FY 2000 to FY 2006. Those hospitals with nurse staffing below 4.0 mean productive medical-surgical RN and registry hours per patient day in FY 2000 had a significantly larger, positive change in mean productive hours than did hospitals with average mean productive hours (approximately 5 hours) in FY 2000. Hospitals staffing above 6.0 mean productive hours per patient day in FY 2000 changed their staffing less than did hospitals with average mean productive hours.


Systematic review of respiratory health among dairy workers.
The dairy industry is changing on a global scale with larger, more efficient operations. The impact of this change on worker health and safety, specifically, associations between occupational lung disease and inhalation exposures, has yet to be reported in a comprehensive review of the scientific literature. Therefore, a three-tier process was used to identify information using a keyword search of online databases of scientific literature. Of the 147 citations reviewed, 52 met initial screening criteria, and 30 were included in this review. Dairy workers experience lung conditions such as asthma, chronic obstructive pulmonary disease, hypersensitivity pneumonitis, chronic bronchitis, and cancer. Recent pulmonary function studies have identified obstructive lung changes among dairy farm workers. The increased scale of dairy production with significant changes in technology and work practices has altered inhalation exposure patterns among dairy workers. The inhalation exposure in the dairy work environment may elicit differing inflammatory responses in relation to timing of initial exposure as well as to repeated exposures. Few studies have measured inhalation exposure while simultaneously assessing the impact of the exposure on lung function of dairy farm workers. Even fewer studies have been implemented to assess the impact of aerosol control technology to reduce inhalation exposure. Future research should evaluate worker exposure to aerosols through a task-based approach while utilizing novel methods to assess inhalation exposure and associated inflammatory responses. Finally, potential solutions should be developed and tested to reduce inhalation exposure to inflammatory agents and respiratory diseases in the dairy farm work environment.


Infrared thermography for monitoring severity and treatment of diabetic foot infections.
Monitoring of diabetic foot infections is largely based on clinical assessment, which is limited by moderate reliability. We conducted a prospective study to explore monitoring of thermal asymmetry (difference between mean plantar temperature of the affected and unaffected foot) for the assessment of severity of diabetic foot infections. In patients with moderate or severe diabetic foot infections (International Working Group on the Diabetic Foot infection-grades 3 or 4) we measured thermal asymmetry with an advanced infrared thermography setup during the first 4-5 days of in-hospital treatment, in addition to clinical assessments and tests of serum inflammatory markers (white blood cell counts and C-reactive protein levels). We assessed the change in thermal asymmetry from baseline to final assessment, and investigated its association with infection-grades and serum inflammatory markers. In seven included patients, thermal asymmetry decreased from median 1.8°C (range: -0.6 to 8.4) at baseline to 1.5°C (range: -0.1 to 5.1) at final assessment (P = 0.515). In three patients who improved to infection-grade 2, thermal asymmetry at baseline (median 1.6°C (range: -0.6 to 1.6)) and final assessment (1.5°C (range: 0.4 to 5.1)) remained similar (P = 0.302). In four patients who did not improve to infection-grade 2, thermal asymmetry decreased from median 4.3°C (range: 1.8 to 8.4) to 1.9°C (range: -0.1 to 4.4; P = 0.221). No correlations were found between thermal asymmetry and infection-grades (r = -0.347; P = 0.445), CRP-levels (r = 0.321; P = 0.482) or WBC (r = -0.250; P = 0.589) during the first 4-5 days of hospitalization. Based on these explorative findings we suggest that infrared thermography is of no value for monitoring diabetic foot infections during in-hospital treatment.


Replacing dietary palmitic acid with elaidic acid (t-C18:1 delta9) depresses HDL and increases CETP activity in cebus monkeys.
The question whether dietary trans fatty acids affect lipoprotein metabolism similarly to specific saturated fatty acids was investigated in 11 normolipemic cebus monkeys by exchanging 5% dietary energy (%en) between elaidic (t-C18:1 delta9) and palmitic acid (16:0) in two test diets (30%en fat + 100 mg cholesterol/1000 kcal diet) conforming to the American Heart Association (AHA) Step 1 guidelines. These were compared with a normal control diet rich in saturated fat and cholesterol (38%en fat + 180 mg cholesterol/1000 kcal diet). The control diet was fed initially for 14 wk, followed by each of the the two test diets in a crossover design. Plasma lipid concentrations were determined four times between the 6th and 14th wk. Turnover studies (using 125I-HDL and 131I-LDL) were conducted after 9 wk in each dietary period. Relative to the control diet, both test diets significantly reduced plasma total cholesterol (TC), HDL cholesterol (HDL-C) and VLDL plus LDL cholesterol (LDL-C) concentrations; triglyceride (TG) concentrations tended to be lower. However, the trans diet resulted in a significantly greater reduction in HDL-C than the palmitate diet (124 +/- 17, 117 +/- 18 and 106 +/- 13 mg/dL for the control, palmitate and trans diets, respectively). The palmitate diet significantly decreased the TC/HDL-C ratio by 11% when compared with the control diet (1.68 +/- 0.17 vs. 1.89 +/- 0.30), whereas the trans diet had no effect (1.81 +/- 0.20 vs. 1.89 +/- 0.30). Kinetic studies revealed that, relative to the control diet, both test diets significantly lowered the LDL apolipoprotein B (apoB) pool size, principally reflecting an increase in the LDL apoB fractional catabolic rate (FCR) related to the reduced cholesterol intake. Between the two test diets, no significant differences in LDL kinetic parameters were observed. Both test diets significantly decreased HDL apoA1 concentrations in comparison with the control diet, which was partly explained by an increase in the fractional catabolic rate of HDL. Of the two test diets, the trans diet was associated with a 9.5% greater HDL FCR than the palmitate diet (P < 0.08) and a significant increase in plasma cholesteryl ester transfer protein (CETP) activity (% transfer 114 +/- 7 vs. 91 +/- 7; P < 0.03). Thus, palmitic acid- and elaidic acid-rich diets produced identical effects on LDL metabolism in normocholesterolemic cebus monkeys fed diets with low levels of cholesterol, whereas elaidic acid depressed HDL-C, attributable to both increased CETP activity and HDL clearance.


Patient perceptions of surgical informed consent: is repeat back helpful or harmful?
Informed consent (IC)comprehension is suboptimal. Repeat back (RB)-asking the patient to repeat in their own words key elements of the consent-is believed to improve the consent process. This study aims to assess the impact of RB on patient perceptions of surgical informed consent. Secondary analysis of a randomized trial. Elective surgical patients were consented using iMedConsent, the VA's computer-based IC platform. Patients were randomized to RB (IC could not be signed until the patient satisfactorily expressed key elements) or standard iMedConsent (no RB). A questionnaire was given immediately after IC assessing time for decision, satisfaction with and ease of understanding consent, and the amount of information provided about the proposed surgery (e.g., indications, benefits, risks, and alternatives). Groups were compared with Χ tests. We enrolled 575 subjects (276 RB and 299 no RB); 92% were men with a mean age of 61.6 years and high school level reading ability. The groups were comparable in their perceptions regarding time to make a decision (RB 88% Strongly Agree (SA), no RB 88% SA; P = 0.61), satisfaction with consent (RB 90% SA, no RB 87% SA; P = 0.27), ease of understanding (RB 69% SA, no RB 67% SA; P = 0.73) receipt of the right amount of information regarding the indications (RB 85% SA, no RB 87%; P = 0.61), the benefits (RB 87% SA, no RB 86% SA; P = 0.29), and the risks (RB 87% SA, no RB 84% SA; P = 0.19) of surgery. More of the RB group felt they received the right amount of information about alternatives to surgery (RB 80% SA) than did the no RB group (69% SA); P = 0.01. Patients were highly satisfied with RB during surgical IC RB is not detrimental to the consent process and may improve informed consent for surgery. Clinical Trials Identifier NCT00288899 http://www.clinicaltrials.gov.


Incident dementia in a defined older Chinese population.
Current knowledge about incident dementia is mainly derived from studies undertaken in the West, showing that dementia is related to older age, low socio-economic status, lack of social network, depression and cardiovascular disease risk factors. We know little about incidence and predictors of dementia in China, where the prevalence is increasing and the patterns of risk factors are different. Using a standard interview method, we examined 1526 non-demented people aged ≥65 years who had at least minimal educational level in China in a 7.5-year follow up. Incident dementia was diagnosed by GMS-AGECAT algorithms and psychiatrists. Age-standardised incidence of dementia was 14.7 per 1000 person-years (95%CI 11.3-18.2 per 1000 person-years). The increased risk was significantly associated with age, female gender (adjusted odds ratio 2.48, 95%CI 1.20-5.13), low educational levels, smoking, angina (2.58, 1.01-6.59) and living with fewer family members. Among participants with low educational level, the increased risk was associated with higher income, and with the highest and lowest occupational classes; adjusted odds ratio 2.74 (95%CI 1.12-6.70) for officers/teachers, 3.11 (1.61-6.01) for manual labourers/peasants. Our findings of high incidence of dementia and increased risk among people having low education levels but high income suggest a more potential epidemic and burden of dementia populations in China. Maintaining social network and activities and reducing cardiovascular factors in late life could be integrated into current multi-faceted preventive strategies for curbing the epidemic of dementia.


Antigenic variation in the phase I lipopolysaccharide of Coxiella burnetii isolates.
Coxiella burnetii isolates from a variety of clinical and geographical sources were screened for antigenic variation of lipopolysaccharides (LPSs) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis coupled with silver staining or immunoblotting. All isolates from chronic Q fever or other sources possessed a phase I-type LPS. These LPSs appeared to fall into three groups based on the sodium dodecyl sulfate-polyacrylamide gel electrophoresis profile or on reactivity with rabbit anti-C. burnetti antisera. The LPS of one group was identified on isolates from milk, ticks, or primary Q fever. The two remaining groups were found almost exclusively on isolates from human cases of chronic Q fever.


The influence of the type of anaesthesia on postoperative pain after kidney transplantation.
The postoperative pain treatment is one of important factors of a successful outcome after kidney transplantation. Improperly controlled pain leads to agitation, tachycardia, hypertension and increases risk of respiratory complications. Many studies have demonstrated good analgetic effect of morphine delivered by the method of patient controlled analgesia (PCA). Because the intensity of postoperative pain in end-stage kidney insufficiency patients can be modified by the type of received anaesthesia, it was decided to analyze the influence of standardized general anaesthesia on postoperative morphine consumption. 140 (ASA III) patients scheduled for kidney transplantation were included. Patients were divided into four groups; group K (control)--anaesthetised with fentanyl and N2O, group 1--fentanyl, N2O plus halothane, group 2--fentanyl, N2O plus propofol, group 3--fentanyl, N2O plus isoflurane. After operation and initial loading dose, PCA infusion of morphine was started. Bolus doses were set to 30 ug/kg, and lockout interval 10 min. Our results suggest that observed greater morphine consumption after GA with the use of propofol is connected with better psychomotor functions. In that group patients were better oriented and more efficiently controlled the PCA pump and pain.


Biological hierarchies and the concept of homology.
Although most biologists agree that homology must be defined in terms of common ancestry, the details of this definition remain controversial. We review briefly the disagreements concerning the formal definition of homology and the methodology used to establish specific cases of homology. Our principal focus, however, is a third area of disagreement: whether morphological characters can be homologous even if their developmental and genetic bases are not homologous, and whether behavioral characters can be homologous even if their morphological substrates are not homologous. We contend that attempts to reduce behavioral homology to morphological homologies, and morphological homology to genetic and developmental homologies, are misguided and based on a failure to recognize the hierarchical nature of biological organization. Genes, developmental processes, morphological structures, physiological functions and behaviors all constitute different levels of biological organization. These levels are causally interrelated, but there is no one-to-one correspondence between characters at different levels. Furthermore, the causal relationships between characters at different levels may change during the course of evolution. As a result, higher level characters may be homologous, even though some of their constituent lower level characters are not homologous. In support of this assertion, we provide several examples of homologous morphological characters that are based on non-homologous developmental precursors and processes, and of homologous behavioral characters that are based on non-homologous morphological structures. In allowing one to recognize homologies at any level of organization, independently of homologies at other levels, the hierarchical concept of homology also allows one to ask important questions about how evolutionary changes at the various levels of organization are related to one another.


Human atrial conduction with reference to heart rate and refractory periods.
Atrial conduction time (ACT) between a stimulating pacemaker electrode and an "impulse-detecting" monophasic action potential (MAP) electrode has been determined in 20 healthy males. A small increase in ACT was found with increasing paced heart rates. Using the extra stimulus technique, ACT was found to increase when the early ectopics approached the atrial effective refractory period (AERP). The degree of ACT prolongation was similar at long and short basic ACTs, indicating that the slowing of impulse propagation probably occurred near the simulating electrode and presumably within 1/2-1 cm from that electrode. The increase in ACT close to the AERP was of importance when determining the atrioventricular refractory period. In three recordings a supernormal phase of conduction was found. The small errors of the MAP recordings in detecting the time of excitation make the method sensitive enough to detect small differences in ACT.


Density fitting of intramonomer correlation effects in symmetry-adapted perturbation theory.
Symmetry-adapted perturbation theory (SAPT) offers insight into the nature of intermolecular interactions. In addition, accurate energies can be obtained from the wave function-based variant of SAPT provided that intramonomer electron correlation effects are included. We apply density-fitting (DF) approximations to the intramonomer correlation corrections in SAPT. The introduction of this approximation leads to an improvement in the computational cost of SAPT by reducing the scaling of certain SAPT terms, reducing the amount of disk I/O, and avoiding the explicit computation of certain types of MO integrals. We have implemented all the intramonomer correlation corrections to SAPT through second-order under the DF approximation. Additionally, leading third-order terms are also implemented. The accuracy of this truncation of SAPT is tested against the S22 test set of Hobza and co-workers [Phys. Chem. Chem. Phys. 8, 1985 (2006)]. When the intramonomer corrections to dispersion are included in SAPT, a mean absolute deviation of 0.3-0.4 kcal mol(-1) is observed for the S22 test set when using an aug-cc-pVDZ basis. The computations on the adenine-thymine complexes in the S22 test set with an aug-cc-pVDZ basis represent the largest SAPT computations to date that include this degree of intramonomer correlation. Computations of this size can now be performed routinely with our newly developed DF-SAPT program.


Global improvement of vascular function and redox state with low-dose folic acid: implications for folate therapy in patients with coronary artery disease.
Although dietary folate fortification lowers plasma homocysteine and may reduce cardiovascular risk, high-dose folic acid therapy appears to not alter clinical outcome. Folic acid and its principal circulating metabolite, 5-methyltetrahydrofolate, improve vascular function, but mechanisms relating folate dose to vascular function remain unclear. We compared the effects of folic acid on human vessels using pharmacological high-dose versus low-dose treatment, equivalent to dietary folate fortification. Fifty-six non-folate-fortified patients with coronary artery disease were randomized to receive low-dose (400 microg/d) or high-dose (5 mg/d) folic acid or placebo for 7 weeks before coronary artery bypass grafting. Vascular function was quantified by magnetic resonance imaging before and after treatment. Vascular superoxide and nitric oxide bioavailability were determined in segments of saphenous vein and internal mammary artery. Low-dose folic acid increased nitric oxide-mediated endothelium-dependent vasomotor responses, reduced vascular superoxide production, and improved enzymatic coupling of endothelial nitric oxide synthase through availability of the cofactor tetrahydrobiopterin. No further improvement in these parameters occurred with high-dose compared with low-dose treatment. Whereas plasma 5-methyltetrahydrofolate increased proportionately with treatment dose of folic acid, vascular tissue 5-methyltetrahydrofolate showed no further increment with high-dose compared with low-dose folic acid. Low-dose folic acid treatment, comparable to daily intake and dietary fortification, improves vascular function through effects on endothelial nitric oxide synthase and vascular oxidative stress. High-dose folic acid treatment provides no additional benefit. These direct vascular effects are related to vascular tissue levels of 5-methyltetrahydrofolate rather than plasma levels. High-dose folic acid treatment likely confers no further benefit in subjects already receiving folate supplementation.


Review: neuroestrogen regulation of socio-sexual behavior of males.
It is thought that estrogen (neuroestrogen) synthesized by the action of aromatase in the brain from testosterone activates male socio-sexual behaviors, such as aggression and sexual behavior in birds. We recently found that gonadotropin-inhibitory hormone (GnIH), a hypothalamic neuropeptide, inhibits socio-sexual behaviors of male quail by directly activating aromatase and increasing neuroestrogen synthesis in the preoptic area (POA). The POA is thought to be the most critical site of aromatization and neuroestrogen action for the regulation of socio-sexual behavior of male birds. We concluded that GnIH inhibits socio-sexual behaviors of male quail by increasing neuroestrogen concentration beyond its optimal concentration in the brain for expression of socio-sexual behavior. On the other hand, it has been reported that dopamine and glutamate, which stimulate male socio-sexual behavior in birds and mammals, inhibit the activity of aromatase in the POA. Multiple studies also report that the activity of aromatase or neuroestrogen is negatively correlated with changes in male socio-sexual behavior in fish, birds, and mammals including humans. Here, we review previous studies that investigated the role of neuroestrogen in the regulation of male socio-sexual behavior and reconsider the hypothesis that neuroestrogen activates male socio-sexual behavior in vertebrates. It is considered that basal concentration of neuroestrogen is required for the maintenance of male socio-sexual behavior but higher concentration of neuroestrogen may inhibit male socio-sexual behavior.


Carbapenem-Resistant Enterobacterales in Long-Term Care Facilities: A Global and Narrative Review.
The emergence of carbapenem-resistant Enterobacterales (CRE) has become a major public health concern. Moreover, its colonization among residents of long-term care facilities (LTCFs) is associated with subsequent infections and mortality. To further explore the various aspects concerning CRE in LTCFs, we conducted a literature review on CRE colonization and/or infections in long-term care facilities. The prevalence and incidence of CRE acquisition among residents of LTCFs, especially in California, central Italy, Spain, Japan, and Taiwan, were determined. There was a significant predominance of CRE in LTCFs, especially in high-acuity LTCFs with mechanical ventilation, and thus may serve as outbreak centers. The prevalence rate of CRE in LTCFs was significantly higher than that in acute care settings and the community, which indicated that LTCFs are a vital reservoir for CRE. The detailed species and genomic analyses of CRE among LTCFs reported that Klebsiella pneumoniae is the primary species in the LTCFs in the United States, Spain, and Taiwan. KPC-2-containing K. pneumoniae strains with sequence type 258 is the most common sequence type of KPC-producing K. pneumoniae in the LTCFs in the United States. IMP-11- and IMP-6-producing CRE were commonly reported among LTCFs in Japan. OXA-48 was the predominant carbapenemase among LTCFs in Spain. Multiple risk factors associated with the increased risk for CRE acquisition in LTCFs were found, such as comorbidities, immunosuppressive status, dependent functional status, usage of gastrointestinal devices or indwelling catheters, mechanical ventilation, prior antibiotic exposures, and previous culture reports. A high CRE acquisition rate and prolonged CRE carriage duration after colonization were found among residents in LTCFs. Moreover, the patients from LTCFs who were colonized or infected with CRE had poor clinical outcomes, with a mortality rate of up to 75% in infected patients. Infection prevention and control measures to reduce CRE in LTCFs is important, and could possibly be controlled via active surveillance, contact precautions, cohort staffing, daily chlorhexidine bathing, healthcare-worker education, and hand-hygiene adherence.


Phosphorylation-independent effects of second messenger system modulators on gamma-aminobutyric acidA receptor complex function.
Recent studies investigating the functional significance of gamma-aminobutyric acidA (GABAA) receptor complex phosphorylation have employed membrane-permeant compounds to manipulate second messenger systems. Although these compounds affect GABAA receptor function, the dependence of these effects on phosphorylation has not been established. Here we report that several second messenger system modulations can decrease GABAA receptor function independently of their effects on protein phosphorylation. Brain membrane vesicles were lysed and resealed in the presence of EDTA to chelate internal Mg2+. Under these conditions, phosphorylation of vesicle proteins was almost completely inhibited, as determined by incorporation of 32P into phosphoproteins. In these lysed/resealed vesicles, an inhibition of muscimol-stimulated 36Cl- uptake was observed with the cAMP analogs 8-(4-chlorophenylthio)-cAMP, N6,O2'-dibutyryl-cAMP, and 8-bromo-cAMP, the protein kinase inhibitor H7, and the adenylate cyclase activator forskolin. In both intact and EDTA-treated lysed/resealed microsacs, cAMP analogs and H7 inhibited binding of the GABAA receptor ligand [3H]SR 95531 at concentrations shown to inhibit muscimol-stimulated 36Cl- uptake. Forskolin was observed to inhibit the binding of t-butylbicyclophosphoro-[35S]thionate, a ligand that binds to a site on the chloride channel. These results demonstrate that compounds commonly used to alter second messenger systems affect the receptor sites and function of the GABAA receptor chloride channel by mechanisms that do not involve protein phosphorylation. In light of these findings, results obtained with these compounds should be interpreted with caution.


Individual differences in language ability are related to variation in word recognition, not speech perception: evidence from eye movements.
The authors examined speech perception deficits associated with individual differences in language ability, contrasting auditory, phonological, or lexical accounts by asking whether lexical competition is differentially sensitive to fine-grained acoustic variation. Adolescents with a range of language abilities (N = 74, including 35 impaired) participated in an experiment based on McMurray, Tanenhaus, and Aslin (2002). Participants heard tokens from six 9-step voice onset time (VOT) continua spanning 2 words (beach/peach, beak/peak, etc.) while viewing a screen containing pictures of those words and 2 unrelated objects. Participants selected the referent while eye movements to each picture were monitored as a measure of lexical activation. Fixations were examined as a function of both VOT and language ability. Eye movements were sensitive to within-category VOT differences: As VOT approached the boundary, listeners made more fixations to the competing word. This did not interact with language ability, suggesting that language impairment is not associated with differential auditory sensitivity or phonetic categorization. Listeners with poorer language skills showed heightened competitors fixations overall, suggesting a deficit in lexical processes. Language impairment may be better characterized by a deficit in lexical competition (inability to suppress competing words), rather than differences in phonological categorization or auditory abilities.


Feedback-induced polarization switching in a Nd-doped fiber laser.
We demonstrate a switching effect between the two polarization eigenstates of a Nd-doped fiber laser when it is subjected to an optical feedback signal. Conditions for an optimization of this effect are recognized to depend on the relative orientation of the fiber eigenaxes and the polarization states of the pump and feedback beams.


Improving the utility of evidence synthesis for decision makers in the face of insufficient evidence.
To identify and suggest strategies to make insufficient evidence ratings in systematic reviews more actionable. A workgroup comprising members from the Evidence-Based Practice (EPC) Program of the Agency for Healthcare Research and Quality convened throughout 2020. We conducted iterative discussions considering information from three data sources: a literature review for relevant publications and frameworks, a review of a convenience sample of past systematic reviews conducted by the EPCs, and an audit of methods used in past EPC technical briefs. We identified five strategies for supplementing systematic review findings when evidence on benefits or harms is expected to be, or found to be, insufficient: 1) reconsider eligible study designs, 2) summarize indirect evidence, 3) summarize contextual and implementation evidence, 4) consider modelling, and 5) incorporate unpublished health system data in the evidence synthesis. While these strategies may not increase the strength of evidence, they may improve the utility of reports for decision makers. Adopting these strategies depends on feasibility, timeline, funding, and expertise of the systematic reviewers. Throughout the process of evidence synthesis of early scoping, protocol development, review conduct, and review presentation, authors can consider these five strategies to supplement evidence with insufficient rating to make it more actionable for end-users.


Low-level laser (light) therapy (LLLT) in skin: stimulating, healing, restoring.
Low-level laser (light) therapy (LLLT) is a fast-growing technology used to treat a multitude of conditions that require stimulation of healing, relief of pain and inflammation, and restoration of function. Although skin is naturally exposed to light more than any other organ, it still responds well to red and near-infrared wavelengths. The photons are absorbed by mitochondrial chromophores in skin cells. Consequently, electron transport, adenosine triphosphate nitric oxide release, blood flow, reactive oxygen species increase, and diverse signaling pathways are activated. Stem cells can be activated, allowing increased tissue repair and healing. In dermatology, LLLT has beneficial effects on wrinkles, acne scars, hypertrophic scars, and healing of burns. LLLT can reduce UV damage both as a treatment and as a prophylactic measure. In pigmentary disorders such as vitiligo, LLLT can increase pigmentation by stimulating melanocyte proliferation and reduce depigmentation by inhibiting autoimmunity. Inflammatory diseases such as psoriasis and acne can also be managed. The noninvasive nature and almost complete absence of side effects encourage further testing in dermatology.


Is vanadium a biometal for boreal cyanolichens?
Molybdenum (Mo) nitrogenase has long been considered the predominant isoenzyme responsible for dinitrogen fixation worldwide. Recent findings have challenged the paradigm of Mo hegemony, and highlighted the role of alternative nitrogenases, such as the vanadium-nitrogenase. Here, we first characterized homeostasis of vanadium (V) along with other metals in situ in the dinitrogen fixing cyanolichen Peltigera aphthosa. These lichens were sampled in natural sites exposed to various levels of atmospheric metal deposition. These results were compared with laboratory experiments where Anabaena variabilis, which is also hosting the V-nitrogenase, and a relatively close relative of the lichen cyanobiont Nostoc, was subjected to various levels of V. We report here that V is preferentially allocated to cephalodia, specialized structures where dinitrogen fixation occurs in tri-membered lichens. This specific allocation is biologically controlled and tightly regulated. Vanadium homeostasis in lichen cephalodia exposed to various V concentrations is comparable to the one observed in Anabaena variabilis and other dinitrogen fixing organisms using V-nitrogenase. Overall, our findings support current hypotheses that V could be a more important factor in mediating nitrogen input in high latitude ecosystems than previously recognized. They invite the reassessment of current theoretical models linking metal dynamics and dinitrogen fixation in boreal and subarctic ecosystems.


Mental health and psychosocial problems in the aftermath of the Nepal earthquakes: findings from a representative cluster sample survey.
Two large earthquakes in 2015 caused widespread destruction in Nepal. This study aimed to examine frequency of common mental health and psychosocial problems and their correlates following the earthquakes. A stratified multi-stage cluster sampling design was employed to randomly select 513 participants (aged 16 and above) from three earthquake-affected districts in Nepal: Kathmandu, Gorkha and Sindhupalchowk, 4 months after the second earthquake. Outcomes were selected based on qualitative preparatory research and included symptoms of depression and anxiety (Hopkins Symptom Checklist-25); post-traumatic stress disorder (PTSD Checklist-Civilian); hazardous alcohol use (AUDIT-C); symptoms indicating severe psychological distress (WHO-UNHCR Assessment Schedule of Serious Symptoms in Humanitarian Settings (WASSS)); suicidal ideation (Composite International Diagnostic Interview); perceived needs (Humanitarian Emergency Settings Perceived Needs Scale (HESPER)); and functional impairment (locally developed scale). A substantial percentage of participants scored above validated cut-off scores for depression (34.3%, 95% CI 28.4-40.4) and anxiety (33.8%, 95% CI 27.6-40.6). Hazardous alcohol use was reported by 20.4% (95% CI 17.1-24.3) and 10.9% (95% CI 8.8-13.5) reported suicidal ideation. Forty-two percent reported that 'distress' was a serious problem in their community. Anger that was out of control (symptom from the WASSS) was reported by 33.7% (95% CI 29.5-38.2). Fewer people had elevated rates of PTSD symptoms above a validated cut-off score (5.2%, 95% CI 3.9-6.8), and levels of functional impairment were also relatively low. Correlates of elevated symptom scores were female gender, lower caste and greater number of perceived needs. Residing in Gorkha and Sindhupalchowk districts and lower caste were also associated with greater perceived needs. Higher levels of impaired functioning were associated with greater odds of depression and anxiety symptoms; impaired functioning was less strongly associated with PTSD symptoms. Four months after the earthquakes in Nepal, one out of three adults experienced symptoms of depression and distressing levels of anger, one out of five engaged in hazardous drinking, and one out of ten had suicidal thoughts. However, posttraumatic stress symptoms and functional impairment were comparatively less frequent. Taken together, the findings suggest that there were significant levels of psychological distress but likely low levels of disorder. The findings highlight the importance of indicated prevention strategies to reduce the risk of distress progressing to disorder within post-disaster mental health systems of care.


Humming greatly increases nasal nitric oxide.
The paranasal sinuses are major producers of nitric oxide (NO). We hypothesized that oscillating airflow produced by humming would enhance sinus ventilation and thereby increase nasal NO levels. Ten healthy subjects took part in the study. Nasal NO was measured with a chemiluminescence technique during humming and quiet single-breath exhalations at a fixed flow rate. NO increased 15-fold during humming compared with quiet exhalation. In a two-compartment model of the nose and sinus, oscillating airflow caused a dramatic increase in gas exchange between the cavities. Obstruction of the sinus ostium is a central event in the pathogenesis of sinusitis. Nasal NO measurements during humming may be a useful noninvasive test of sinus NO production and ostial patency. In addition, any therapeutic effects of the improved sinus ventilation caused by humming should be investigated.


The relation between common sleep problems and emotional and behavioral problems among 2- and 3-year-olds in the context of known risk factors for psychopathology.
The contribution of sleep problems to emotional and behavioral problems among young children within the context of known risk factors for psychopathology was examined. Data on 2- and 3-year-olds, representative of Canadian children without a chronic illness, from three cross-sectional cohorts of the Canadian National Longitudinal Study of Child and Youth were analysed (n = 2996, 2822, and 3050). The person most knowledgeable (PMK), usually the mother, provided information about her child, herself, and her family. Predictors included: child health status and temperament; parenting and PMK depressive symptomatology; family demographics (e.g., marital status, income) and functioning. Child sleep problems included night waking and bedtime resistance. Both internalizing/emotional (i.e., anxiety) and externalizing/behavioral problems (i.e., hyperactivity, aggression) were examined. Adjusting for other known risk factors, child sleep problems accounted for a small, but significant, independent proportion of the variance in internalizing and externalizing problems. Structural equation models examining the pathways linking risk factors to sleep problems and emotional and behavioral problems were a good fit of the data. Results were replicated on two additional cross-sectional samples. The relation between sleep problems and emotional and behavioral problems is independent of other commonly identified risk factors. Among young children, sleep problems are as strong a correlate of child emotional and behavioral problems as PMK depressive symptomatology, a well-established risk factor for child psychopathology. Adverse parenting and PMK symptomatology, along with difficult temperament all contribute to both sleep problems and emotional and behavioral problems. Children's sleep problems appear to exacerbate emotional and behavioral problems.


Biogenic amines in cultured neuroblastoma and astrocytoma cells.
The presence of biogenic amines in cultured cells of mouse neuroblastoma C-1300 (clone NB-2a) was suggested by fluorescence-microscope histochemistry. Incubation in media containing L-[(14)C]tyrosine and L-[(14)C]tryptophan for 24 h, followed by high-voltage electrophoresis, radiochromatogram scanning, and scintillation counting, confirmed the presence of [(14)C]dopamine, [(14)C]norepinephrine, [(14)C]epinephrine, [(14)C]serotonin, [(14)C]tyramine, and [(14)C]octopamine. Dopamine, norepinephrine, epinephrine, and serotonin were demonstrated spectrophotofluorometrically in concentrations, expressed as micrograms amine per milligram protein, of 1.19, 0.027, 0.038, and 0.148, respectively, for cells in a stationary growth phase. Fluorescence-microscope histochemistry also suggested the presence of biogenic amines in cultured astrocytoma cells (cell line C6). Spectrophotofluorometric assay of cells in a stationary growth phase demonstrated intracellular dopamine, norepinephrine, epinephrine, and serotonin in concentrations significantly lower than those of neuroblastoma cells.