Evidence of West Nile Virus (WNV) Circulation in Wild Birds and WNV RNA Negativity in Mosquitoes of the Danube Delta Biosphere Reserve, Romania, 2016.
West Nile virus (WNV) is a zoonotic flavivirus whose transmission cycle in nature includes wild birds as amplifying hosts and ornithophilic mosquito vectors. Bridge vectors can transmit WNV to mammal species potentially causing West Nile Fever. Wild bird migration is a mode of WNV introduction into new areas. The Danube Delta Biosphere Reserve (DDBR) is a major stopover of wild birds migrating between Europe and Africa. The aim of this study was to investigate the presence of WNV in the DDBR during the 2016 transmission season in wild birds and mosquitoes. Blood from 68 wild birds (nine different species) trapped at four different locations was analyzed by competitive ELISA and Virus Neutralization Test (VNT), revealing positive results in 8/68 (11.8%) of the wild birds by ELISA of which six samples (three from juvenile birds) were confirmed seropositive by VNT. Mosquitoes (n = 6523, 5 genera) were trapped with CDC Mini Light traps at two locations and in one location resting mosquitoes were caught. The presence of WNV RNA was tested in 134 pools by reverse transcription quantitative PCR (RT-qPCR). None of the pools was positive for WNV-specific RNA. Based on the obtained results, WNV was circulating in the DDBR during 2016.


Selective Constraints on Coding Sequences of Nervous System Genes Are a Major Determinant of Duplicate Gene Retention in Vertebrates.
The evolutionary history of vertebrates is marked by three ancient whole-genome duplications: two successive rounds in the ancestor of vertebrates, and a third one specific to teleost fishes. Biased loss of most duplicates enriched the genome for specific genes, such as slow evolving genes, but this selective retention process is not well understood. To understand what drives the long-term preservation of duplicate genes, we characterized duplicated genes in terms of their expression patterns. We used a new method of expression enrichment analysis, TopAnat, applied to in situ hybridization data from thousands of genes from zebrafish and mouse. We showed that the presence of expression in the nervous system is a good predictor of a higher rate of retention of duplicate genes after whole-genome duplication. Further analyses suggest that purifying selection against the toxic effects of misfolded or misinteracting proteins, which is particularly strong in nonrenewing neural tissues, likely constrains the evolution of coding sequences of nervous system genes, leading indirectly to the preservation of duplicate genes after whole-genome duplication. Whole-genome duplications thus greatly contributed to the expansion of the toolkit of genes available for the evolution of profound novelties of the nervous system at the base of the vertebrate radiation.


Cascade of atopic dermatitis comorbidities in children after birth for 15 years.
Individuals with atopic dermatitis often develop other conditions. This study aimed to determine how atopic dermatitis comorbidities develop in children over time. This population-based administrative cohort study used national health insurance data. We traced individuals born in Korea between 2002 and 2003 to 2018. The date of initial atopic dermatitis diagnosis was set as the index date. Fifty-three childhood comorbidities of atopic dermatitis were identified as outcomes of interest by performing a comprehensive literature search and comparing the prevalence of diagnostic codes in children with and without atopic dermatitis. Four control children per individual in the atopic dermatitis group were randomly matched based on sex and index date. The association between atopic dermatitis and the development of each specified disease was assessed using proportional hazard assumption, followed by mapping of the temporal sequences of interconnected comorbidities. The atopic dermatitis and control groups contained 67,632 and 270,528 individuals, respectively. The median age at the index date was 10 months, whereas the median follow-up period was 15 years. Twenty diseases that were associated with a higher risk of atopic dermatitis were identified and a chain of interconnected conditions created. The progression began in childhood with febrile seizures, constipation, and asthma, and was later associated with the emergence of food allergy, allergic rhinitis, psychiatric disorders, and autoimmune diseases. Our study highlights the temporal nature of atopic dermatitis comorbidities in children, and indicates that an understanding of the comorbidities may inform its clinical management and treatment.


Knowledge and attitude toward waterpipe smoking: scale validation and correlates in the Lebanese adult population.
The World Health Organization declared waterpipe smoking as a growing public health concern. Our study aimed to assess waterpipe smoking knowledge and attitude among current waterpipe smokers in comparison with nonsmokers and to evaluate its association with current and cumulative waterpipe smoking. A cross-sectional study was carried between May and December 2018 using a multistage cluster sample across Lebanon. Nonsmokers or exclusive current waterpipe smokers were eligible to participate. A total of 4046 adult individuals participated in the study, 38.3% (n = 1550) being current waterpipe smokers. Current waterpipe smoking was positively associated with female gender [adjusted odds ratio (aOR) 1.79, 95%CI 1.40-2.28] and having a mother who is a regular smoker (aOR 1.47, 95%CI 1.15-1.88) while it was negatively associated with better knowledge about waterpipe smoking harm (aOR 0.95, 95%CI 0.93-0.97) and positive attitude toward waterpipe smoking ban (aOR 0.98, 95%CI 0.96-0.99). Cumulative waterpipe smoking was negatively associated with female gender (β = - 0.012), higher education level (β = - 0.28), and positive attitude toward WPS ban (β = - 0.06). Waterpipe smoking is largely present among the Lebanese population, with waterpipe smokers showing lower knowledge about harm and more intolerant attitude about banning. Health awareness supported by media campaigns and governmental initiatives should be conducted to educate people and change their perceptions regarding waterpipe smoking.


Coherence between low-frequency activation of the motor cortex and tremor in patients with essential tremor.
In healthy people, rhythmic activation of the motor cortex in the 15-30 Hz frequency range accompanies and contributes to voluntarily-generated postural contractions of contralateral muscle. In patients with Parkinson's disease, an abnormal low-frequency activation of the motor areas of the cortex occurs and has been directly linked to the characteristic 3-6 Hz rest tremor of this disease. We therefore investigated whether the motor cortex is involved in the transmission of the rhythmic motor drive responsible for generating essential tremor. Non-invasive recordings of activity from the hand area of the motor cortex were made from six patients with essential tremor by magnetoencephalography. The recordings were made simultaneously with the electromyogram recorded from contralateral finger muscles during periods of postural tremor. A statistical spectral analysis was done to determine at which frequencies the two signals were correlated. Spectral analysis of the electromyogram signals showed a significant low-frequency component at the frequency of the tremor bursts. However, there was no coherence between magnetoencephalogram and electromyogram recordings at the tremor frequency, indicating that no correlation existed between the tremor signal and low-frequency activity recorded from the primary motor cortex in individuals with essential tremor. Coherence at frequencies higher than the tremor frequency was similar to that in healthy individuals performing voluntary postural contractions. The absence of significant coherence between the magnetoencephalogram and electromyogram at tremor frequencies suggests that in essential tremor the tremor is imposed on the active muscle through descending pathways other than those originating in the primary motor cortex. These findings challenge the model widely used to explain the efficacy of neurosurgical treatment of essential tremor, are in contrast to those of previous studies of parkinsonian rest tremor, and highlight an important difference in the pathophysiology of essential and parkinsonian tremor.


Error analysis for Mueller matrix measurement.
The linear errors of Mueller matrix measurements are formulated for misalignment, depolarization, and incorrect retardation of the polarimetric components. The measured errors of a Mueller matrix depend not only on the imperfections of the measuring system but also on the Mueller matrix itself. The error matrices for different polarimetric systems are derived and also evaluated for the straight-through case. The error matrix for a polarizer-sample-analyzer system is much simpler than those for more complicated systems. The general error matrix is applied to null ellipsometry, and the obtained errors in ellipsometric parameters psi and delta are identical to the errors specifically derived for null ellipsometry with depolarization.


Biopositive effects of low-dose UVB on epidermis: coordinate upregulation of antimicrobial peptides and permeability barrier reinforcement.
Whereas high-dose ultraviolet B (UVB) is detrimental to the epidermal permeability barrier, suberythemal doses of UVB are used to treat atopic dermatitis (AD), which is characterized by defective permeability barrier and antimicrobial function. As epidermal permeability barrier and antimicrobial peptide (AMP) expression are coregulated and interdependent functions, we hypothesized that suberythemal doses of UVB exposure could regulate AMP expression in parallel with permeability barrier function. Hairless mice were exposed to 40 mJ cm(-2) UVB (about 1/2 minimal erythema dose) daily for 1 or 3 days. Twenty-four hours after the last exposure, epidermal barrier function was assessed and skin specimens were taken for western blotting, immunohistochemistry, and quantitative reverse transcription-PCR for mouse beta-defensin (mBD)-2, mBD3 and cathelin-related antimicrobial peptide (CRAMP). mRNA levels of the vitamin D receptor (VDR), 1alpha-hydroxylase and key epidermal lipid synthetic enzymes were also quantified. After 3 days of UVB exposure, acceleration of barrier recovery and augmentation in expression of epidermal differentiation markers (for example, involucrin and filaggrin) occurred in parallel with increased mBD2, mBD3, and CRAMP expression at both the mRNA and protein level. VDR, 1alpha-hydroxylase, and the major epidermal lipid synthetic enzymes were also upregulated. When an inhibitor of 1alpha, 25 dihydroxyvitamin D(3) formation, ketoconazole, was applied immediately after UVB exposure, the cutaneous vitamin D system was inhibited, which in turn blocked epidermal lipid synthesis, AMP expression, and permeability barrier homeostasis, suggesting that the beneficial effect of low-dose UVB depends, at least in part, on activation of the cutaneous vitamin D system. Our results provide new insights into the mechanisms whereby low-dose UVB comprises effective therapy for AD.


Selection of aquatic microbiota exposed to the herbicides flufenacet and metazachlor.
Herbicides are important, ubiquitous environmental contaminants, but little is known about their interaction with bacterial aquatic communities. Here, we sampled a protected natural freshwater habitat and characterised its microbiome in interaction with herbicides. We evolved the freshwater microbiomes in a microcosm assay of exposure (28 days) to flufenacet and metazachlor at environmental concentrations of 0.5, 5 and 50 μg L-1 . Inhibitory effects of herbicides were exemplarily assessed in cultured bacteria from the same pond (Pseudomonas alcaligenes, Paenibacillus amylolyticus and Microbacterium hominis). Findings were compared to long-term concentrations as provided by local authorities. Here, environmental concentrations reached up to 11 μg L-1 (flufenacet) and 76 μg L-1 (metazachlor). Bacteria were inhibited at minimum inhibitory concentrations far above these values; however, concentrations of 50 μg L-1 of flufenacet resulted in measurable growth impairment. While most herbicide-exposed microcosm assays did not differ from controls, Acidobacteria were selected at high environmental concentrations of herbicides. Alpha-diversity (e.g., taxonomic richness on phylum level) was reduced when aquatic microbiomes were exposed to 50 μg metazachlor or flufenacet. One environmental strain of P. alcaligenes showed resistance to high concentrations of flufenacet (50 g L-1 ). In total, this study reveals that ecologic imbalance due to herbicide use significantly impacts aquatic microbiomes.


[Features of homeostasis in surgeons].
The article deals with results of studies concerning homeostasis system in surgeons working in major industrial city. Findings are disorders of elements balance, that manifest through lower content of some essential elements and accumulation of toxic ones. The authors defined correlations between adaptational level, immunologic and elemental states of the body.


Gender dependence of serum soluble Klotho in acromegaly.
In acromegaly, disease activity is biochemically assessed by growth hormone (GH) and insulin-like growth factor-1 (IGF-1) levels. However, they are often discrepant, as several factors including gender influence their relationship. We recently found excessively high serum levels of soluble Klotho (sKl) in acromegalic patients, which depended on GH to a comparable extent as IGF-1. To further elucidate the relationship between GH and sKl, we examined the effect of gender on sKl in patients with untreated acromegaly. We determined GH, IGF-1 and sKl in sera of 62 consecutive patients with newly diagnosed acromegaly (31 females/31 males, aged 20-85 years). For their given GH excess at presentation with acromegaly, females had lower IGF-1 (490 ± 33 vs 604 ± 33 ng/ml, P = 0·02), but higher sKl [5171 ± 590 vs 3439 ± 431 pg/ml (mean ± SE), P = 0·02] levels than males. In multiple regression analysis, IGF-1 was closely associated with logGH (estimate 139, SE 47, P = 0·005) and BMI (estimate 14·2, SE 4·8, P = 0·005). sKl was closely associated with logGH (estimate 3088, SE 652, P = 0·0001) and gender (estimate 2034, SE 612, P = 0·002), and to a lesser extent with BMI (estimate 174, SE 66, P = 0·01). For a given GH status, sKl concentrations are higher and IGF-1 concentrations are lower in women than in men. GH is the strongest predictor for both sKl and IGF-1, but gender needs to be considered when using these parameters for monitoring acromegalic patients.


Altered crystal violet permeability and lytic behavior in antibiotic-resistant and -sensitive mutants of Neisseria gonorrhoeae.
Wild-type, antibiotic-resistant and hypersensitive isogenic strains of Neisseria gonorrhoeae were studied for uptake of crystal violet, rates of autolysis, and response to lysozyme. Total uptake of crystal violet was similar in all strains at 0 C but varied significantly at 37 C. Mutation at the nonspecific resistance locus ery resulted in relative impermeability to crystal violet at 37 C, as compared to wild type. The penetration barrier to crystal violet at 37 C was overcome by addition of 5 mM ethylenediaminetetraacetic acid. Mutation at ery also resulted in reduced rates of autolysis and reduced sensitivity to high concentrations of lysozyme under conditions of divalent cation (Mg2+) depletion. In contrast, mutation at the nonspecific drug hypersensitivity locus env resulted in increased uptake of crystal violet at 37 C, due to increased binding of dye to crude envelope as well as increased penetration into cytoplasm. The env mutants were also more rapidly autolytic and more sensitive to lysozyme than wild type in the absence of Mg2+. These results suggest that the cell envelopes of ery mutants are more stable and less permeable and those of env mutants are less stable and more permeable than wild-type strains.


Major deletions in the gene encoding the p53 tumor antigen cause lack of p53 expression in HL-60 cells.
The tumor antigen p53 is overproduced in transformed cells of various species, including man. HL-60 is an exceptional human tumor cell line that does not express this protein. Hybridization of polyadenylylated mRNA of these cells with a human p53 cDNA probe (p53-H14), which we cloned, had indicated a total absence of the mature-size (3.0 kilobases) or any aberrant p53 mRNA species. Analysis of the genomic HL-60 DNA indicated that the p53 gene in these cells was significantly altered. Most of the gene was deleted, and the residual p53 sequences of these cells, which show weak homology, mapped to the corresponding 5' region of the p53 gene. In agreement with previously documented results, we found that HL-60 cells have an amplified c-myc gene. We suggest that the deficiency of the p53 protein in HL-60 cells could have been overcome by using an alternative metabolic pathway. The c-myc product is a candidate for such an alternative protein.


What the Hel: recent advances in understanding rifampicin resistance in bacteria.
Rifampicin is a clinically important antibiotic that binds to, and blocks the DNA/RNA channel of bacterial RNA polymerase (RNAP). Stalled, nonfunctional RNAPs can be removed from DNA by HelD proteins; this is important for maintenance of genome integrity. Recently, it was reported that HelD proteins from high G+C Actinobacteria, called HelR, are able to dissociate rifampicin-stalled RNAPs from DNA and provide rifampicin resistance. This is achieved by the ability of HelR proteins to dissociate rifampicin from RNAP. The HelR-mediated mechanism of rifampicin resistance is discussed here, and the roles of HelD/HelR in the transcriptional cycle are outlined. Moreover, the possibility that the structurally similar HelD proteins from low G+C Firmicutes may be also involved in rifampicin resistance is explored. Finally, the discovery of the involvement of HelR in rifampicin resistance provides a blueprint for analogous studies to reveal novel mechanisms of bacterial antibiotic resistance.


Informal Caregiving and Network Turnover Among Older Adults.
Personal network turnover, a combination of lost and added network contacts, is suggested to affect health as well as moderate access to social support and resources. This article tests whether the caregiving process is associated with network turnover in later life and whether the process is different for men and women. Network turnover was assessed using two waves of personal network data from the National Social Life, Health, and Aging Project. Network contacts were uniquely identified in each wave making it possible to document contact loss and addition. Rates of change were modeled using Poisson regression. Respondents transitioning into caregiving lost and added network contacts at higher rates than non-caregivers. Conversely, respondents providing care during both waves and respondents transitioning out of the role saw no significant levels of network turnover. The analysis provided minimal evidence of gender differences. Findings suggest that the initial shift into the caregiving role is associated with notable personal network change. This is an important consideration given that long-term network instability may lead to poor health and limited access to social resources whereas adaptive network change tends to elicit more positive outcomes.


Clonal variability in CD95 expression is the major determinant in Fas-medicated, but not chemotherapy-medicated apoptosis in the RPMI 8226 multiple myeloma cell line.
CD95 (Fas/APO-1) is a member of the TNFR superfamily that induces apoptosis following cross-linking with its cognate ligand, CD95L (FasL/APO-1L) or agonist antibody. The human myeloma cell line, RPMI 8226, has limited sensitivity to CD95-mediated apoptosis, with a maximum of 65% of the population responding. To determine the source of the limited sensitivity to CD95-mediated apoptosis, we isolated multiple clones from the RPMI-8226 cell line by limiting dilution. Analysis of these clones demonstrated that sensitivity to CD95-mediated cell death directly correlated with CD95 expression. Clones with high levels of CD95 expression had greater than 90% cell death, whereas cells with low levels of expression had less than 10% cell death. In contrast, no correlative differences were identified for other members of the DISC complex, or for members of the anti-apoptotic Bcl-2 family. We further examined the sensitivity of the 8226 clones to various cytotoxic agents. Although modest clonal variability was demonstrated in response to the chemotherapeutic drugs, doxorubicin, etoposide (VP-16), and vincristine, there was no correlation between CD95 function and sensitivity to chemotherapeutic drugs. These results indicate that in this cell line, receptor expression is rate limiting in CD95-mediated apoptosis, whereas CD95 expression was not a determinant in drug-induced programmed cell death.


Isolation of an FMRP-associated messenger ribonucleoprotein particle and identification of nucleolin and the fragile X-related proteins as components of the complex.
The loss of FMR1 expression due to trinucleotide repeat expansion leads to fragile X syndrome, a cause of mental retardation. The encoded protein, FMRP, is a member of a gene family that also contains the fragile X-related proteins, FXR1P and FXR2P. FMRP has been shown to be a nucleocytoplasmic shuttling protein that selectively binds a subset of mRNAs, forms messenger ribonucleoprotein (mRNP) complexes, and associates with translating ribosomes. Here we describe a cell culture system from which we can isolate epitope-tagged FMRP along with mRNA, including its own message, and at least six other proteins. We identify two of these proteins as FXR1P and FXR2P by using specific antisera and identify a third protein as nucleolin by using mass spectrometry. The presence of nucleolin is confirmed by both reactivity with a specific antiserum as well as reverse coimmunoprecipitation where antinucleolin antiserum immunoprecipitates endogenous FMRP from both cultured cells and mouse brain. The identification of nucleolin, a known component of other mRNPs, adds a new dimension to the analysis of FMRP function, and the approach described should also allow the identification of the remaining unknown proteins of this FMRP-associated mRNP as well as the other bound mRNAs.


mRNA trans-splicing dual AAV vectors for (epi)genome editing and gene therapy.
Large genes including several CRISPR-Cas modules like gene activators (CRISPRa) require dual adeno-associated viral (AAV) vectors for an efficient in vivo delivery and expression. Current dual AAV vector approaches have important limitations, e.g., low reconstitution efficiency, production of alien proteins, or low flexibility in split site selection. Here, we present a dual AAV vector technology based on reconstitution via mRNA trans-splicing (REVeRT). REVeRT is flexible in split site selection and can efficiently reconstitute different split genes in numerous in vitro models, in human organoids, and in vivo. Furthermore, REVeRT can functionally reconstitute a CRISPRa module targeting genes in various mouse tissues and organs in single or multiplexed approaches upon different routes of administration. Finally, REVeRT enabled the reconstitution of full-length ABCA4 after intravitreal injection in a mouse model of Stargardt disease. Due to its flexibility and efficiency REVeRT harbors great potential for basic research and clinical applications.


Plasma somatostatin levels in patients with chronic idiopathic intestinal pseudo-obstruction.
Somatostatin participates in the control of gut motility. Recently, somatostatin analogs have been proposed as therapeutic agents for chronic intestinal pseudo-obstruction, although the endogenous somatostatin pattern has never been assessed in this syndrome. We aimed to evaluate fasting and postprandial plasma somatostatin levels in patients with chronic idiopathic intestinal pseudo-obstruction (CIIP). We studied eight patients with CIIP and 10 healthy volunteers. Blood samples were taken at regular intervals while patients and subjects fasted and during the 3 h after a standard solid/liquid meal (550 kcal) had been eaten. Somatostatin was measured by radioimmunoassay. Fasting somatostatin levels were normal, whereas postprandial peptide responses were markedly impaired or even absent in patients with CIIP. An impaired postprandial somatostatin response in patients with CIIP seems to be characteristic of this heterogenous disorder. Whether the lack of somatostatin response to a meal identifies patients with severe gut dysmotility for whom treatment with somatostatin analogs would be useful remains to be verified.


A microfluidic platform for evaporation-based salt screening of pharmaceutical parent compounds.
We describe a microfluidic platform to screen for salt forms of pharmaceutical compounds (PCs) via controlled evaporation. The platform enables on-chip combinatorial mixing of PC and salt former solutions in a 24-well array (~200 nL/well), which is a drastic reduction in the amount of PC needed per condition screened compared to traditional screening approaches that require ~100 μL/well. The reduced sample needs enable salt screening at a much earlier stage in the drug development process, when only limited quantities of PCs are available. Compatibility with (i) solvents commonly used in the pharmaceutical industry, and (ii) Raman spectroscopy for solid form identification was ensured by using a hybrid microfluidic platform. A thin layer of elastomeric PDMS was utilized to retain pneumatic valving capabilities. This layer is sandwiched between layers of cyclic-olefin copolymer, a material with low air and solvent permeability and low Raman background to yield a physically rigid and Raman compatible chip. A solvent-impermeable thiolene layer patterned with evaporation channels permits control over the rate of solvent evaporation. Control over the rate of solvent evaporation (2-15 nL h(-1)) results in consistent, known rates of increase in the supersaturation levels attained on-chip, and increases the probability for crystalline solids to form. The modular nature of the platform enables on-chip Raman and birefringence analysis of the solid forms. Model compounds, tamoxifen and ephedrine, were used to validate the platform's ability to screen for salts. On-chip Raman analysis helped to identify six different salts each of tamoxifen and ephedrine.


Evaluating Classifier Confidence for Surface EMG Pattern Recognition.
Surface electromyogram (EMG) can be employed as an interface signal for various devices and software via pattern recognition. In EMG-based pattern recognition, the classifier should not only be accurate, but also output an appropriate confidence (i.e., probability of correctness) for its prediction. If the confidence accurately reflects the likelihood of true correctness, then it will be useful in various application tasks, such as motion rejection and online adaptation. The aim of this paper is to identify the types of classifiers that provide higher accuracy and better confidence in EMG pattern recognition. We evaluate the performance of various discriminative and generative classifiers on four EMG datasets, both visually and quantitatively. The analysis results show that while a discriminative classifier based on a deep neural network exhibits high accuracy, it outputs a confidence that differs from true probabilities. By contrast, a scale mixture model-based classifier, which is a generative classifier that can account for uncertainty in EMG variance, exhibits superior performance in terms of both accuracy and confidence.


Frequency of Giardia lamblia among children in Dohuk, northern Iraq.
Out of 1261 stool specimens collected from children in Dohuk city, northern Iraq, the prevalence of Giardia lamblia infection was 38.5%. The highest rate of infection was in orphan care centres (48.1%) and the lowest in the paediatric hospital (31.3%). The age group 10-12 years had the highest rate (81.2%) and 7-9 years the lowest (22.9%); boys had a higher rate than girls. Some infected samples (70/486) showed double or triple infections and G. lamblia was combined with Hymenolepis nana, Blastocystis hominis, Entamoeba histolytica and Iodamoeba buetschlii.


A comparison of patient teaching outcomes among postoperative coronary artery bypass graft (CABG) patients.
Shortened hospitalizations following cardiac surgery necessitate re-evaluation of how pertinent information for self-care management and reduction of coronary artery disease risk factors can be incorporated into an effective inpatient cardiac patient teaching program. This study investigated the effect of three different teaching approaches (i.e., an inpatient teaching program, a postdischarge telephone follow-up program, and a postdischarge group teaching program) among 90 patients who had undergone coronary artery bypass graft surgery. Teaching outcomes were evaluated in this study by use of the Heart Disease Management Questionnaire and Cardiac Surgical Patient Teaching Satisfaction Inventory. Analyses of the data revealed similar patient teaching outcomes regardless of the type of teaching intervention the participant received. Findings supported the effectiveness of the inpatient teaching protocol which focused on "survival skills" for self-care management postdischarge. The findings are important in the redesign of teaching programs which are efficient yet meaningful to the patient within today's health care environment. Additional findings indicated that patients who had longer lengths of stay and those with more vessels bypassed were the least satisfied with their teaching. Further implications for nurses involved in patient teaching are discussed.


Study of the effects of salicylic acid on soybean mitochondrial lipids and respiratory properties using the alternative oxidase as a stress-reporter protein.
Biotic and abiotic stresses can lead to modifications in the lipid composition of cell membranes. Although mitochondria appear to be implicated in stress responses, little is known about the membrane lipid changes that occur in these organelles in plants. Besides cytochrome c oxidase, plant mitochondria have an alternative oxidase (AOX) that accepts electrons directly from ubiquinol, dissipating energy as heat. AOX upregulation occurs under a variety of stresses and its induction by salicylic acid (SA) has been observed in different plant species. AOX was also suggested to be used as a functional marker for cell reprogramming under stress. In the present study, we have used etiolated soybean (Glycine max (L.) Merr. cv Cresir) seedlings to study the effects of SA treatment on the lipid composition and the respiratory properties of hypocotyl mitochondria. AOX expression was studied in detail, as a reporter protein, to evaluate whether modifications in mitochondrial energy metabolism were occurring. In mitochondria extracted from SA-treated seedlings, AOX capacity and protein contents increased. Both AOX1 and AOX2b transcripts accumulated in response to SA, but with different kinetics. A reduction in external NADH oxidation capacity was observed, whereas succinate respiration remained unchanged. The phospholipid composition of mitochondria remained similar in control and SA-treated plants, but a reduction in the relative amount of linolenic acid was observed in phosphatidylcholine, phosphatidylethanolamine and cardiolipin. The possible causes of the fatty acid modifications observed, and the implications for mitochondrial metabolism are discussed.


Immunomodulation of acute experimental spinal cord injury with human immunoglobulin G.
Immunomodulation of acute spinal cord injury may inhibit the activity of specific inflammatory cascades and result in recovery of motor function. In this study, evaluation of the protective effect of a well-known anti-inflammatory immunomodulator, immunoglobulin G (IgG), was conducted in rats after a 50 g/cm contusion spinal cord injury. Following injury, 400 mg/kg of IgG was administered to the treatment group. Twenty-four hours later, animals were assessed functionally via an inclined plane and the Basso-Beattie-Bresnahan motor scale and compared to controls. Tissue was reviewed for myeloperoxidase activiy (MPO) and lipid peroxidation (LPO), and electron microscopy was conducted to assess tissue ultrastructure. Significant functional preservation was observed in the IgG treatment group. In addition, biochemical assays revealed decreased MPO activity, and electron microscopic views of tissue showed preserved ultrastructure. IgG treatment following acute contusion injury to the rat spinal cord confers functional and structural neuroprotection.


Physicochemical studies on turnip-yellow-mosaic virus. Homogeneity, relative molecular masses, hydrodynamic radii and concentration-dependence of parameters in non-dissociating solvents.
Turnip-yellow-mosaic virus, with its stable, highly spherical and monodisperse character, was chosen as a suitable model substance with which to test hydrodynamic theories of transport. Sedimentation coefficients, diffusion coefficients (obtained through photon correlation spectroscopy) and viscosities were measured accurately as a function of concentration in well-defined and nearly neutral buffer systems. Ancillary information was also obtained from very-low-speed sedimentation-equilibrium experiments. The coefficients expressing the variation in sedimentation and diffusion coefficients with weight concentration were obtained, and by combination with other data it was possible to avoid assumptions concerning solvation and transform such regression coefficients into the form appropriate to volume fractions. Some measure of support for Batchelor's [(1972) J. Fluid Mech. 52, 245-268] calculations was thus obtained, but over most of the pH range the coefficients were significantly smaller than those calculated from his theory. It seems likely that electrostatic interactions are responsible for the discrepancies. Hydrodynamic radii (from diffusion coefficients) were in very fair agreement with those calculated from the thermodynamic excluded-volume term, but were higher than indicated by electron microscopy and X-ray diffraction, a discrepancy ascribable to solvation.


Transcatheter Aortic Valve Implantation Under Angiographic Guidance With and Without Adjunctive Transesophageal Echocardiography.
Although transcatheter aortic valve implantation (TAVI) is still currently guided by transesophageal echocardiography (TEE) in a considerable number of hospitals, exclusive angiographic (Angio) guidance seems a reasonable approach in this setting. To date, however, no studies have directly compared the outcomes of TAVI according to the imaging modality used for procedural guidance. We, therefore, used data from a large multicenter data repository to compare the outcomes of TAVI guided exclusively by Angio and ATEE. All consecutive patients with severe aortic stenosis who underwent TAVI with the CoreValve Revalving System (CRS) in 9 Italian centers from September 2007 to March 2014, dichotomized according to the imaging support used to guide the procedure (ATEE and Angio), were included. Thirty-day and 12-month clinical outcomes were evaluated. Propensity matching analysis was performed to adjust for baseline differences. A total of 625 patients were included (256 and 369 patients were included in the ATEE and Angio groups, respectively). Patients from the ATEE more frequently underwent TAVI under general anesthesia compared with Angio group (37.9% vs 22.8%, respectively, p <0.001). Importantly, ∼80% of the patients experienced mild or even less aortic regurgitation as assessed by angiography after the procedure, without between-group differences. Postdilation and valve-in-valve rates were equivalent (24.7% vs 25%, p = 0.934 and 5.5% vs 3.4%, respectively, p = 0.217). No differences were revealed in the rates of death, cardiovascular death, and stroke or transient ischemic attack at 12-month follow-up. These results were sustained after propensity matching analysis. In conclusion, as long as a comprehensive procedural planning is performed, TAVI with CRS may be performed exclusively under angiographic guidance without the need for associated TEE.


Ischemic nephropathy: more than a simple renal artery narrowing.
Renal artery stenosis in elderly patients is mainly caused by atherosclerosis. The prevalence of this disorder in patients with chronic kidney diseases is reported to be 0.5% to 5.5%. However, because the patients with atherosclerotic renal artery disease are mostly asymptomatic, the true prevalence is expected to be higher. Renovascular hypertension and ischemic nephropathy are two main consequences of this disease, but it is difficult to determine in which patient the progress of stenosis may cause these syndromes. The big challenge in renal artery stenosis is how to manage the patients. In the past 70 years, it has been believed that simply maintaining of kidney perfusion by opening the stenosis could control blood pressure and preserve kidney function. Nowadays, the blood pressure can be controlled well by medical treatment without the need for revascularization; however, management of ischemic nephropathy remains a dilemma. With advancements in understanding the pathophysiology of changes in the parenchyma of the kidney after stenosis, it is now generally accepted that only a minority of patients with ischemic nephropathy will benefit from revascularization. Nonetheless, finding these patients is critical and need more randomized trials to show who mostly benefit from revascularization and when it may save the kidney.


A statistical framework to assess cross-frequency coupling while accounting for confounding analysis effects.
Cross frequency coupling (CFC) is emerging as a fundamental feature of brain activity, correlated with brain function and dysfunction. Many different types of CFC have been identified through application of numerous data analysis methods, each developed to characterize a specific CFC type. Choosing an inappropriate method weakens statistical power and introduces opportunities for confounding effects. To address this, we propose a statistical modeling framework to estimate high frequency amplitude as a function of both the low frequency amplitude and low frequency phase; the result is a measure of phase-amplitude coupling that accounts for changes in the low frequency amplitude. We show in simulations that the proposed method successfully detects CFC between the low frequency phase or amplitude and the high frequency amplitude, and outperforms an existing method in biologically-motivated examples. Applying the method to in vivo data, we illustrate examples of CFC during a seizure and in response to electrical stimuli.


Scalable high-fidelity growth of semiconductor nanorod arrays with controlled geometry for photovoltaic devices using block copolymers.
Controlled density semiconducting oxide arrays are highly desirable for matching nanometer length scales specific to emerging applications. This work demonstrates a facile one-step method for templating hydrothermal growth which provides arrays with high-fidelity tuning of nanorod spacing and diameter. This solution-based method leverages the selective swelling of block copolymer micelle templates, which can be rationally designed by tuning molecular weight and volume fraction.


The von Hippel-Lindau tumor suppressor gene inhibits hepatocyte growth factor/scatter factor-induced invasion and branching morphogenesis in renal carcinoma cells.
Loss of function in the von Hippel-Lindau (VHL) tumor suppressor gene occurs in familial and most sporadic renal cell carcinomas (RCCs). VHL has been linked to the regulation of cell cycle cessation (G(0)) and to control of expression of various mRNAs such as for vascular endothelial growth factor. RCC cells express the Met receptor tyrosine kinase, and Met mediates invasion and branching morphogenesis in many cell types in response to hepatocyte growth factor/scatter factor (HGF/SF). We examined the HGF/SF responsiveness of RCC cells containing endogenous mutated (mut) forms of the VHL protein (VHL-negative RCC) with that of isogenic cells expressing exogenous wild-type (wt) VHL (VHL-positive RCC). We found that VHL-negative 786-0 and UOK-101 RCC cells were highly invasive through growth factor-reduced (GFR) Matrigel-coated filters and exhibited an extensive branching morphogenesis phenotype in response to HGF/SF in the three-dimensional (3D) GFR Matrigel cultures. In contrast, the phenotypes of A498 VHL-negative RCC cells were weaker, and isogenic RCC cells ectopically expressing wt VHL did not respond at all. We found that all VHL-negative RCC cells expressed reduced levels of tissue inhibitor of metalloproteinase 2 (TIMP-2) relative to the wt VHL-positive cells, implicating VHL in the regulation of this molecule. However, consistent with the more invasive phenotype of the 786-0 and UOK-101 VHL-negative RCC cells, the levels of TIMP-1 and TIMP-2 were reduced and levels of the matrix metalloproteinases 2 and 9 were elevated compared to the noninvasive VHL-positive RCC cells. Moreover, recombinant TIMPs completely blocked HGF/SF-mediated branching morphogenesis, while neutralizing antibodies to the TIMPs stimulated HGF/SF-mediated invasion in vitro. Thus, the loss of the VHL tumor suppressor gene is central to changes that control tissue invasiveness, and a more invasive phenotype requires additional genetic changes seen in some but not all RCC lines. These studies also demonstrate a synergy between the loss of VHL function and Met signaling.


TNNI3K mutation in familial syndrome of conduction system disease, atrial tachyarrhythmia and dilated cardiomyopathy.
Locus mapping has uncovered diverse etiologies for familial atrial fibrillation (AF), dilated cardiomyopathy (DCM), and mixed cardiac phenotype syndromes, yet the molecular basis for these disorders remains idiopathic in most cases. Whole-exome sequencing (WES) provides a powerful new tool for familial disease gene discovery. Here, synergistic application of these genomic strategies identified the pathogenic mutation in a familial syndrome of atrial tachyarrhythmia, conduction system disease (CSD), and DCM vulnerability. Seven members of a three-generation family exhibited the variably expressed phenotype, three of whom manifested CSD and clinically significant arrhythmia in childhood. Genome-wide linkage analysis mapped two equally plausible loci to chromosomes 1p3 and 13q12. Variants from WES of two affected cousins were filtered for rare, predicted-deleterious, positional variants, revealing an unreported heterozygous missense mutation disrupting the highly conserved kinase domain in TNNI3K. The G526D substitution in troponin I interacting kinase, with the most deleterious SIFT and Polyphen2 scores possible, resulted in abnormal peptide aggregation in vitro and in silico docking models predicted altered yet energetically favorable wild-type mutant dimerization. Ventricular tissue from a mutation carrier displayed histopathological hallmarks of DCM and reduced TNNI3K protein staining with unique amorphous nuclear and sarcoplasmic inclusions. In conclusion, mutation of TNNI3K, encoding a heart-specific kinase previously shown to modulate cardiac conduction and myocardial function in mice, underlies a familial syndrome of electrical and myopathic heart disease. The identified substitution causes a TNNI3K aggregation defect and protein deficiency, implicating a dominant-negative loss of function disease mechanism.


TET1 deficiency attenuates the DNA damage response and promotes resistance to DNA damaging agents.
Recent studies have shown that loss of TET1 may play a significant role in the formation of tumors. Because genomic instability is a hallmark of cancer, we examined the potential involvement of 10-11 translocation 1 (TET1) in the DNA damage response (DDR). Here we demonstrate that, in response to clinically relevant doses of ionizing radiation (IR), human glial cells made TET1-deficient with lentiviral vectors displayed greater numbers of colony forming units and lower levels of apoptotic markers compared with glial cells transduced with control vectors; yet, they harbored greater DNA strand breaks. The G2/M check point and expression of cyclin B1 were greatly diminished in TET1-deficient cells, and TET1-deficient cells displayed lower levels of γH2A.x following exposure to IR. Levels of DNA-PKcs, which are DNA-PK complex members, were lower in TET1-deficient cells compared with control cell lines. However, levels of ATM were similar in both cell lines. Cyclin B1, DNA-PKcs, and γH2A.x levels were each rescued by reintroduction of the TET1 catalytic domain. Finally, cytosine methylation within intron 1 of PRKDC, the gene encoding DNA-PKcs, was significantly higher upon depletion of TET1. Taken together, this study illustrates the involvement of TET1 in the different arms of the DDR and suggests its loss results in the continued survival of cells with genomic instability.


X chromosome evolution in the suni and eland antelope: detection of homologous regions by fluorescence in situ hybridization and G-banding.
Comparative cytogenetics and fluorescence in situ hybridization (FISH) were used to track structural rearrangements in the X chromosomes of two African antelope species, the eland (Taurotragus oryx; tribe Tragelaphini) and the suni (Neotragus moschatus; tribe Neotragini). Using two microdissected cattle chromosome painting probes (one specific for Xp-containing sequences corresponding to Xp24-->p12 of the cattle X, and one specific for Xq-containing sequences corresponding to Xq12-->qter), we show that intrachromosomal rearrangements distinguish the X chromosomes of these species. Furthermore, there is clear evidence that, superimposed on this background of intrachromosomal evolutionary change, the sequences contained in the cattle Xp painting probe appear to have moved as a complete unit during the repatterning of the bovid X. Although we are unable to infer the order of the rearranged chromosomal segments, given the large regions of homology detected by the chromosome paints, we nonetheless believe that this approach (combining conventional and molecular cytogenetic studies on the X chromosome) will prove useful for inferring phylogenetic relationships in the family Bovidae, particularly as more probes become available.


[Ten-year activities of the Faculty for Training of Scientific and Pedagogical Personnel, I.M. Sechenov Moscow Medical Academy].
With the assistance and direct participation of the Presidium of the Russian Academy of Medical Sciences, the I. M. Sechenov Moscow Medical Academy has developed and introduced an unique elitist educational system to train of scientific and pedagogical staff at the pre- and postdiploma training stages on the basis of their close relationships and continuity, within whose framework a specially designed faculty has been successfully operating for 10 years. The basic principle that ensures its effective activity and solution of many didactic problems is the individualization and differentiation of an educational process by using a great diversity of reserves and organizational forms. The created system is unique in Russian and foreign practice. It improves the quality of scientific and pedagogical staff training, promotes more effective creation of staff reserves for the departments of higher educational establishments and research institutes of the Russian Academy of Medical Sciences and the Ministry of Health of the Russian Federation.


EEG correlated functional MRI and postoperative outcome in focal epilepsy.
The main challenge in assessing patients with epilepsy for resective surgery is localising seizure onset. Frequently, identification of the irritative and seizure onset zones requires invasive EEG. EEG correlated functional MRI (EEG-fMRI) is a novel imaging technique which may provide localising information with regard to these regions. In patients with focal epilepsy, interictal epileptiform discharge (IED) correlated blood oxygen dependent level (BOLD) signal changes were observed in approximately 50% of patients in whom IEDs are recorded. In 70%, these are concordant with expected seizure onset defined by non-invasive electroclinical information. Assessment of clinical validity requires post-surgical outcome studies which have, to date, been limited to case reports of correlation with intracranial EEG. The value of EEG-fMRI was assessed in patients with focal epilepsy who subsequently underwent epilepsy surgery, and IED correlated fMRI signal changes were related to the resection area and clinical outcome. Simultaneous EEG-fMRI was recorded in 76 patients undergoing presurgical evaluation and the locations of IED correlated preoperative BOLD signal change were compared with the resected area and postoperative outcome. 21 patients had activations with epileptic activity on EEG-fMRI and 10 underwent surgical resection. Seven of 10 patients were seizure free following surgery and the area of maximal BOLD signal change was concordant with resection in six of seven patients. In the remaining three patients, with reduced seizure frequency post-surgically, areas of significant IED correlated BOLD signal change lay outside the resection. 42 of 55 patients who had no IED related activation underwent resection. These results show the potential value of EEG-fMRI in presurgical evaluation.


Cerebellar parameters in developing 15 day old rat pups treated with propylthiouracil in comparison with 5 and 24 day old.
To investigate and quantify structural parameters in the developing cerebellum during hypothyroidism in pre and postnatal stages in 15 day old rat pups in comparison with 5 and 24 day old. Propylthiouracil (PTU) was fed to rat dams during mating, pregnancy and nursing and their pups in drinking water. Consequently hypothyroxinemia was induced in the dams and the developing foeti during prenatal period and maintained in the dams and pups. The number of treated and control dams was five in each group. The treated and control pups were eight and eleven respectively. The whole cerebellum was dissected out and routinely processed for histological and morphometric analysis. Structural changes in cerebellum were estimated using "design based" stereological methods. The total volume of cerebellum, intracerebellar nuclei and cerebellar compartments were estimated using Cavalieri Principle. Numerical density of cells was estimated using the disector method and the total cell number was then calculated. In the 15 day pups there was significant reduction (P<0.05) in the mean volumes of cerebellum, internal granular layer, molecular layer, cerebellar cortex, mean ratio of the total volume of intracerebellar nuclei to the cerebellar volume and increased mean volume of external granular layer in treated pup group compared with control. The mean volumes of intracerebellar nuclei and white matter and the mean numerical densities and total numbers of neurons and Purkinje cells in intracerebellar nuclei and cerebellum respectively were nearly equal in control and treated groups. Significant increase (P<0.05) in the mean numerical density and total number of glial cells in treated pups compared with control was observed. There was significant decrease (P<0.05) in the mean neuron/neuroglia ratio in the intracerebellar nuclei, mean numerical density and mean total number of granule cells and reduction in the mean ratio of total number of granule/Purkinje cells in the treated group compared with control. The linear regression comparison for the total volume of the intracerebellar nuclei to total volume of the cerebellum in 5, 15 and 24 day control and treated pups and for the total number of glial cells on the total volume of intracerebellar nuclei in the same were significantly different (P<0.05). Total numbers of neurons and glial cells in the intracerebellar nuclei showed peak values in 15 day pups. Thus PTU-induced hypothyroidism causes variation in quantitative structural parameters in developing cerebellum and disrupts progressive cellular developmental processes. Maintenance of normal T4 and T3 levels during growth and maturation of cerebellum is absolutely essential.


Working in London hospitals: perceptions of place in nursing students' employment considerations.
During the past decade, a distinct body of research has started to investigate the dynamics between nursing and place. However, despite attention being paid to a wide-range of nursing subjects, few studies have engaged with the important topic of labour force recruitment. In this context, this study uses a combined questionnaire (n=650), interview (n=30) and focus group (n=7) survey of London-based students, and investigates the complex mix of experiences and perceptions that result in hospitals having varying degrees of popularity as potential workplaces. The findings suggest experiences and perceptions of institutions-often gained on clinical placements-to be important, particularly relating to feeling valued, the quality of patient care, clinical and educational opportunities and team cohesion. These are often combined with experiences and perceptions of locality, relating to factors such as cost of living, travel considerations and sense of personnel safety. The study demonstrates that place is relevant to employment decision-making on multiple scales from wards to regions. Furthermore, that perceptions of potential workplaces result from engagements with complex mixes of cultural, economic and physical features, many of which are the consequences of management. It is argued that in order to effectively unpack workplaces, geographical research of nursing labour may benefit from researching simultaneously both 'inside' institutions, focusing on their dominant cultures of production and sub-cultures, and 'outside', focusing on their local urban or rural contexts.


High-performance ion-pair chromatography method for simultaneous analysis of alliin, deoxyalliin, allicin and dipeptide precursors in garlic products using multiple mass spectrometry and UV detection.
The quality of garlic and garlic products is usually related to their alliin content and allicin release potential. Until now no analytical method was able to quantify simultaneously allicin, its direct precursor alliin (S-allyl-L-cysteine sulfoxide), SAC (S-allyl-L-cysteine) as well as various dipeptides that apparently serve as storage compounds in garlic. It is well known that all these intermediates are involved in the allicin biosynthetic pathway. A simple and rapid HPLC method suitable for routine analysis was developed using eluents containing an ion-pairing reagent. Particularly, heptanesulfonate as ion-pairing reagent guarantees a sufficient separation between alliin and the more retained dipeptides at very low pH. Allicin was eluted after 18 min on a 150 x 3 mm column. Synthetic reference compounds were characterized by the same chromatographic method using a diode-array UV detector and an ion trap mass spectrometer (electrospray ionization) in the multiple MS mode. In routine analysis of garlic bulbs, powders and other products, the diode-array detector is sufficient for a relevant quantification. Our method has been used in studies to improve the quality of garlic and its derived products.


PDZ scaffolds regulate extracellular vesicle production, composition, and uptake.
Extracellular vesicles (EVs) are membrane-limited organelles mediating cell-to-cell communication in health and disease. EVs are of high medical interest, but their rational use for diagnostics or therapies is restricted by our limited understanding of the molecular mechanisms governing EV biology. Here, we tested whether PDZ proteins, molecular scaffolds that support the formation, transport, and function of signal transduction complexes and that coevolved with multicellularity, may represent important EV regulators. We reveal that the PDZ proteome (ca. 150 proteins in human) establishes a discrete number of direct interactions with the tetraspanins CD9, CD63, and CD81, well-known EV constituents. Strikingly, PDZ proteins interact more extensively with syndecans (SDCs), ubiquitous membrane proteins for which we previously demonstrated an important role in EV biogenesis, loading, and turnover. Nine PDZ proteins were tested in loss-of-function studies. We document that these PDZ proteins regulate both tetraspanins and SDCs, differentially affecting their steady-state levels, subcellular localizations, metabolism, endosomal budding, and accumulations in EVs. Importantly, we also show that PDZ proteins control the levels of heparan sulfate at the cell surface that functions in EV capture. In conclusion, our study establishes that the extensive networking of SDCs, tetraspanins, and PDZ proteins contributes to EV heterogeneity and turnover, highlighting an important piece of the molecular framework governing intracellular trafficking and intercellular communication.


Diagnostic Potential of Multimodal MRI Markers in Atypical Parkinsonian Disorders.
The diagnostic potential of multimodal MRI approaches to discriminate among progressive supranuclear palsy (PSP), Parkinson variant of multiple system atrophy (MSA-P) and Parkinson's disease (PD) has not been well investigated. To identify disease-specific neurodegenerative patterns and evaluate the diagnostic accuracy of dedicated MRI, iron concentration (R2*), microstructural integrity (mean diffusivity; MD and fractional anisotropy; FA) as well as volumes were analyzed in patients with PSP, MSA-P and PD. 3T MRI of 18 PSP and 16 MSA-P patients were compared with 16 PD patients matched for age and disease duration as well as 21 healthy controls. Statistical parametric mapping (SPM) was applied to objectively identify focal MRI changes throughout the whole-brain. Following dimensionality reduction of significant and multiple comparison-corrected SPM clusters through principal component analysis (PCA), stepwise receiver-operating characteristic curve analysis (ROC) was applied to determine the diagnostic potential of multimodal MRI parameters. PCA revealed two components involving multiple regions identified from SPM analysis. The first component was primarily composed of the mean MD value of the thalamus and the mean MD and FA values of the dentatorubrothalamic tract and the corpus callosum. The second component mainly consisted of mean MD and FA values of the middle cerebellar peduncle. ROC analysis showed 92% of PSP patients were differentiated correctly from MSA-P and PD and 80% of MSA-P patients could be distinguished from PD. Multimodal MRI improved the detection of disease-specific neurodegenerative patterns in PSP and MSA-P and highlights its potential to improve the diagnostic accuracy of atypical parkinsonian disorders.


Probability of an Autism Diagnosis by Gestational Age.
Early preterm infants (EPT) (<33 6/7 weeks) are at increased risk for autism spectrum disorders (ASDs) but prevalence estimates vary widely across studies. Furthermore, there are very few studies addressing the association between late preterm (LPT) births (34-36 6/7 weeks) and ASDs. To address the question of whether LPT infants carry the same risk for ASDs as full-term infants, this study aimed to estimate the relative probability of an ASD diagnosis using Bayes rule. A retrospective cohort analysis of 406 children was undertaken to look at gestational age, ASDs, and birth history. The application of Bayes rule was used, given that there is not sufficient information about the joint probabilities related to prematurity and autism. Using the estimated gestational age proportions within ASD diagnosis, plus national estimates of ASDs, probabilities for ASDs within a given gestational age were calculated. Among these 406 children with ASDs, 6.7% were EPT and 10.6% were LPT. In comparison to full term, EPT children are at 1.9 multiplicative increase in risk (95% CI [1.3, 2.5]). While the probability of ASDs for LPT children was higher than that for term, the estimated relative risk of the LPT infants was not statistically significant (95% CI [0.9, 1.5]). EPT infants were significantly more likely to be diagnosed with ASDs compared to their term peers. While the relative probability of ASD diagnosis among children born LPT was not statistically significant in this limited sample, the results indicate a possible elevated risk. A larger cohort is needed to adequately estimate this risk.


Neural activity associated with the passive prediction of ambiguity and risk for aversive events.
In economic decision making, outcomes are described in terms of risk (uncertain outcomes with certain probabilities) and ambiguity (uncertain outcomes with uncertain probabilities). Humans are more averse to ambiguity than to risk, with a distinct neural system suggested as mediating this effect. However, there has been no clear disambiguation of activity related to decisions themselves from perceptual processing of ambiguity. In a functional magnetic resonance imaging (fMRI) experiment, we contrasted ambiguity, defined as a lack of information about outcome probabilities, to risk, where outcome probabilities are known, or ignorance, where outcomes are completely unknown and unknowable. We modified previously learned pavlovian CS+ stimuli such that they became an ambiguous cue and contrasted evoked brain activity both with an unmodified predictive CS+ (risky cue), and a cue that conveyed no information about outcome probabilities (ignorance cue). Compared with risk, ambiguous cues elicited activity in posterior inferior frontal gyrus and posterior parietal cortex during outcome anticipation. Furthermore, a similar set of regions was activated when ambiguous cues were compared with ignorance cues. Thus, regions previously shown to be engaged by decisions about ambiguous rewarding outcomes are also engaged by ambiguous outcome prediction in the context of aversive outcomes. Moreover, activation in these regions was seen even when no actual decision is made. Our findings suggest that these regions subserve a general function of contextual analysis when search for hidden information during outcome anticipation is both necessary and meaningful.


Stereophysicochemical variability plots highlight conserved antigenic areas in Flaviviruses.
Flaviviruses, which include Dengue (DV) and West Nile (WN), mutate in response to immune system pressure. Identifying escape mutants, variant progeny that replicate in the presence of neutralizing antibodies, is a common way to identify functionally important residues of viral proteins. However, the mutations typically occur at variable positions on the viral surface that are not essential for viral replication. Methods are needed to determine the true targets of the neutralizing antibodies. Stereophysicochemical variability plots (SVPs), 3-D images of protein structures colored according to variability, as determined by our PCPMer program, were used to visualize residues conserved in their physical chemical properties (PCPs) near escape mutant positions. The analysis showed 1) that escape mutations in the flavivirus envelope protein are variable residues by our criteria and 2) two escape mutants found at the same position in many flaviviruses sit above clusters of conserved residues from different regions of the linear sequence. Conservation patterns in T-cell epitopes in the NS3- protease suggest a similar mechanism of immune system evasion. The SVPs add another dimension to structurally defining the binding sites of neutralizing antibodies. They provide a useful aid for determining antigenically important regions and designing vaccines.


Symptom differences in moderate to severe IBS patients based on predominant bowel habit.
We sought to determine if irritable bowel syndrome (IBS) patients with different bowel habit predominance differ in self-reported viscerosensory symptoms related to the upper and lower gastrointestinal (GI) tract, somatosensory symptoms, and constitutional functions. Six hundred and twenty-five Rome criteria-positive IBS patients completed a bowel symptom questionnaire (BSQ), psychological symptom checklist (SCL-90), and health status (SF-36). Bowel habit predominance for IBS patients was determined using the Rome criteria for functional constipation (IBS-C; n = 140) and functional diarrhea (IBS-D; n = 216). The BSQ included questions about viscerosensory symptoms of the upper (chest pressure, bloating, fullness, early satiety, nausea) and lower GI tract (bloating, pain, incomplete evacuation), somatosensory symptoms related to the musculoskeletal system (pain in neck/shoulders, lower back/hip, muscles/joints), and constitutional functions (sleep, appetite, libido). Analysis was further conducted between the IBS-C and IBS-D patients, controlling for gender and quality of sleep, and using the Bonferroni correction to control for multiple comparisons. Female gender was more prevalent among IBS-C than IBS-D (77% vs 56.1%, p < 0.01), whereas age did not differ (40.2 +/- 1.2 yr vs 39.5 +/- 1.0 yr). Symptoms referred to the upper GI were more prevalent in IBS-C than IBS-D: early satiety (56.7% vs 33.9%, p < 0.004), fullness (63.2% vs 38.5%, p < 0.05), and a trend for upper bloating (80.3 vs 62.6%). IBS-C patients reported higher severity ratings for lower GI bloating (p < 0.001). IBS-C more commonly reported musculoskeletal symptoms (92.2% vs 75.4%, p < 0.001), as well as impairment in sleep (31.3 vs 17.5%, p < 0.009), appetite (35.0% vs 18.4%, p < 0.015) and sexual function (45.2% vs 33.1%, p < 0.0021). There were no differences in SCL-90 and SF-36 scores. Compared with the IBS-D group, the IBS-C patients show greater prevalence of a wide range of symptoms referred to the upper and lower abdomen, musculoskeletal, and constitutional functions. These findings may be related to differences in autonomic or perceptual responses to visceral and somatic stimuli, and are likely to have implications for treatment responses in the two subgroups.


Cortex-based independent component analysis of fMRI time series.
The cerebral cortex is the main target of analysis in many functional magnetic resonance imaging (fMRI) studies. Since only about 20% of the voxels of a typical fMRI data set lie within the cortex, statistical analysis can be restricted to the subset of the voxels obtained after cortex segmentation. While such restriction does not influence conventional univariate statistical tests, it may have a substantial effect on the performance of multivariate methods. Here, we describe a novel approach for data-driven analysis of single-subject fMRI time series that combines techniques for the segmentation and reconstruction of the cortical surface of the brain and the spatial independent component analysis (sICA) of the functional time courses (TCs). We use the mesh of the white matter/gray matter boundary, automatically reconstructed from high-spatial-resolution anatomical MR images, to limit the sICA decomposition of a coregistered functional time series to those voxels which are within a specified region with respect to the cortical sheet (cortex-based ICA, or cbICA). We illustrate our analysis method in the context of fMRI blocked and event-related experimental designs and in an fMRI experiment with perceptually ambiguous stimulation, in which an a priori specification of the stimulation protocol is not possible. A comparison between cbICA and conventional hypothesis-driven statistical methods shows that cortical surface maps and component TCs blindly obtained with cbICA reliably reflect task-related spatiotemporal activation patterns. Furthermore, the advantages of using cbICA when the specification of a temporal model of the expected hemodynamic response is not straightforward are illustrated and discussed. A comparison between cbICA and anatomically unconstrained ICA reveals that--beside reducing computational demand--the cortex-based approach improves the fitting of the ICA model in the gray matter voxels, the separation of cortical components and the estimation of their TCs, particularly in the case of fMRI data sets with a complex spatiotemporal statistical structure.


Federal audit findings in E/M services: a top 10 survival guide.
Physician practices are under intense scrutiny, led by several aggressive and pervasive federal auditing initiatives. One of the primary targets for these initiatives is the most common physician service provided: the E/M service--with the most prevalent E/M service being the basic office visit. Physicians and practice managers should be aware of the risk areas when documenting, coding, and billing E/M services. This article presents the top 10 areas of risk exposure for such services, and throws in an additional eleventh area for an added preventive measure.


Three-dimensional structure of a Streptomyces sviceus GNAT acetyltransferase with similarity to the C-terminal domain of the human GH84 O-GlcNAcase.
The mammalian O-GlcNAc hydrolysing enzyme O-GlcNAcase (OGA) is a multi-domain protein with glycoside hydrolase activity in the N-terminus and with a C-terminal domain that has low sequence similarity to known acetyltransferases, prompting speculation, albeit controversial, that the C-terminal domain may function as a histone acetyltransferase (HAT). There are currently scarce data available regarding the structure and function of this C-terminal region. Here, a bacterial homologue of the human OGA C-terminal domain, an acetyltransferase protein (accession No. ZP_05014886) from Streptomyces sviceus (SsAT), was cloned and its crystal structure was solved to high resolution. The structure reveals a conserved protein core that has considerable structural homology to the acetyl-CoA (AcCoA) binding site of GCN5-related acetyltransferases (GNATs). Calorimetric data further confirm that SsAT is indeed able to bind AcCoA in solution with micromolar affinity. Detailed structural analysis provided insight into the binding of AcCoA. An acceptor-binding cavity was identified, indicating that the physiological substrate of SsAT may be a small molecule. Consistent with recently published work, the SsAT structure further questions a HAT function for the human OGA domain.


Treatment Outcomes and Lessons Learned From Transilluminated Powered Phlebectomy for Varicose Veins in 1034 Patients.
Transilluminated powered phlebectomy (TIPP) is a minimally invasive technique of varicose vein removal, which combines irrigated illumination with tumescent anesthesia for ablation of superficial varicosities and endoscopic-powered venous resection. The objective of this study was to analyze treatment outcomes of this treatment modality. A retrospective evaluation of prospectively collected data from all patients undergoing TIPP procedure for symptomatic varicose veins during a recent 12-year period was performed. Pertinent patient demographics, disease classification, perioperative complications, quality of life, and treatment outcomes were collected and analyzed. A total of 1167 limbs in 1034 patients (mean age, 52.4 years) were treated during the study period. The mean procedure time was 18.4 ± 8.9 minutes (range, 6.0-82.0 minutes). The mean number of incisions for TIPP procedure was 6.3 ± 3.6. All TIPP procedures were technically successful, and no patient required conversion to hook stab phlebectomy. Fifteen (1.5%) patients developed residual or recurrent varicosities, which were treated with sclerotherapy during the follow-up period. Postoperative complications included hematoma at 2 weeks (5.8%), ecchymosis at 2 weeks (32.9%), saphenous neuropathy (0.3%), cellulitis (1.0%), and skin pigmentation (1.9%). There was no postoperative deep vein thrombosis or mortality. Transilluminated powered phlebectomy is an effective method for varicose vein removal and is associated with high clinical success and excellent cosmetic results. Meticulous technical steps are critical in achieving successful outcomes while minimizing complications. Technical considerations and lessons learned from our experiences are discussed in this report.


Genotype and phenotype patterns of human immunodeficiency virus type 1 resistance to enfuvirtide during long-term treatment.
The human immunodeficiency virus type 1 (HIV-1) fusion inhibitor enfuvirtide has recently been introduced into clinical practice and has exhibited efficient anti-HIV-1 activity in combination with other antiretroviral agents. In the present study, we addressed the effect of long-term treatment with enfuvirtide on the intrahost evolution of HIV-1. The genotype and phenotype patterns and the relative replication capacity (rRC) of enfuvirtide-resistant HIV-1 mutants were evaluated in samples from 11 subjects (7 virological nonresponders and 4 responders) who received the compound for more than 1 year in combination with different regimens. Selection of one or more mutations clustering in a sequence (amino acids 36 to 45) of the gp41 N-terminal heptad repeat was observed in samples from the seven virological nonresponders but not in those from responders. In two subjects who discontinued enfuvirtide, reversion of the resistant genotype was detected within 3 months. Recombinant clones bearing mutated gp41 sequences displayed reduced susceptibilities to enfuvirtide, with the 50% inhibitory concentrations (IC(50)s) ranging from 0.6 to 12.8 microg/ml, whereas the IC(50) for isolates with baseline sequences was 0.013 +/- 0.010 microg/ml. Interestingly, long-term monitoring of resistant variants provided evidence that ongoing adaptation to the drug is paralleled by phenotypic changes. A limited drop in the rRC in the absence of drug was observed for clones from four of the seven nonresponders bearing mutations associated with resistance. Overall, the data indicate that the different genotype patterns associated with a detectable degree of HIV-1 resistance to enfuvirtide generated during long-term treatments are characterized by a substantially low genetic barrier, possible ongoing adaptation with increased degrees of resistance, and limited influence on the viral rRC.


Methotrexate and 5-fluorouracil following tamoxifen and premarin in advanced breast cancer.
Complete remissions in patients with advanced breast cancer using either endocrine therapy or chemotherapy are infrequent. Breast tumors are known to be heterogeneous with respect to estrogen receptor (ER) status, and the low complete remission rate may be secondary to this biochemical heterogeneity. Laboratory experiments using breast cancer cells in long-term tissue culture revealed that tamoxifen is cytotoxic, estrogen stimulates the growth of ER-positive cells and can rescue cells from tamoxifen's effect, and sequential MTX/5-FU is synergistic in rapidly growing breast cancer cells. Based on this, a phase II protocol was designed using tamoxifen, 10 mg p.o. b.i.d. for days 1 to 10. This was followed by Premarin, 0.625 mg p.o. b.i.d., on days 11 to 14. On day 14 the patients were given MTX, 200 mg/m2 i.v., followed in 1 hr by 5-FU, 600 mg/m2 i.v. The patients were rescued with leucovorin, 10 mg/m2 24 hr later. This dose of leucovorin was repeated every 6 hr for six doses. The cycle is repeated every 18 days. Thus far, 35 patients have been entered and 32 are currently evaluable for response. The median age is 57 years. The median Karnofsky performance status was 90. Of the 35 patients, 63% were ER-positive, and 23% had received prior endocrine therapy or chemotherapy; 71% were postmenopausal. One third of the patients had visceral-dominant disease. The overall response rate was 69%; 15 of 32 (47%) attained a complete remission, and 7 of 32 (22%) had a partial remission. Toxicity was minimal. Median nadir white blood cell counts were 5600/cu mm, and median nadir platelet counts were 252,000/cu mm. Nausea and vomiting occurred in 25% of patients, but there was no hair loss, rash, or diarrhea. Mucositis was rare. In summary, this combination hormonal-chemotherapy regimen is highly effective with a high complete remission rate and minimal toxicity.


Calorimetric measurements of energetics of defect interactions in fluorite oxides.
Direct measurement by oxide melt solution calorimetry of energetics of mixing in rare earth and yttrium doped zirconia, hafnia, and ceria systems provides support for spectroscopic and computational studies of the location and clustering of vacancies in these systems. Strongly negative heats of mixing are seen when the vacancy is transferred from being nearest neighbor to Y or RE in the sesquioxide to being nearest neighbor to Zr or Hf in the cubic solid solution. In the absence of such redistribution, small positive enthalpies of mixing are seen in CeO2-YO1.5 and CeO2-REO.15 systems. Strongly positive enthalpies of mixing are seen in CeO2-ZrO2, which has a large difference in cation sizes and no vacancy formation. The system Ce0.8Y0.2O1.9-Zr0.8Y0.2O1.9 shows small positive heats of formation with less destabilization in the Ce-rich region, suggestive of "scavenging" of oxygen vacancies by Zr. The calorimetric data obtained in these studies offer direct comparison with the results of computations on defect clusters and their binding energies.


The Application of ATR-FTIR Spectroscopy and the Reversible DNA Conformation as a Sensor to Test the Effectiveness of Platinum(II) Anticancer Drugs.
Platinum(II) complexes have been found to be effective against cancer cells. Cisplatin curbs cell replication by interacting with the deoxyribonucleic acid (DNA), reducing cell proliferation and eventually leading to cell death. In order to investigate the ability of platinum complexes to affect cancer cells, two examples from the class of polyfluorophenylorganoamidoplatinum(II) complexes were synthesised and tested on isolated DNA. The two compounds trans-[N,N'-bis(2,3,5,6-tetrafluorophenyl)ethane-1,2-diaminato(1-)](2,3,4,5,6-pentafluorobenzoato)(pyridine)platinum(II) (PFB) and trans-[N,N'-bis(2,3,5,6-tetrafluorophenyl)ethane-1,2-diaminato(1-)](2,4,6-trimethylbenzoato)(pyridine)platinum(II) (TMB) were compared with cisplatin through their reaction with DNA. Attenuated Total Reflection Fourier Transform Infrared (ATR-FTIR) spectroscopy was applied to analyse the interaction of the Pt(II) complexes with DNA in the hydrated, dehydrated and rehydrated states. These were compared with control DNA in acetone/water (PFB, TMB) and isotonic saline (cisplatin) under the same conditions. Principle Component Analysis (PCA) was applied to compare the ATR-FTIR spectra of the untreated control DNA with spectra of PFB and TMB treated DNA samples. Disruptions in the conformation of DNA treated with the Pt(II) complexes upon rehydration were mainly observed by monitoring the position of the IR-band around 1711 cm-1 assigned to the DNA base-stacking vibration. Furthermore, other intensity changes in the phosphodiester bands of DNA at ~1234 cm-1 and 1225 cm-1 and shifts in the dianionic phosphodiester vibration at 966 cm-1 were observed. The isolated double stranded DNA (dsDNA) or single stranded DNA (ssDNA) showed different structural changes when incubated with the studied compounds. PCA confirmed PFB had the most dramatic effect by denaturing both dsDNA and ssDNA. Both compounds, along with cisplatin, induced changes in DNA bands at 1711, 1088, 1051 and 966 cm-1 indicative of DNA conformation changes. The ability to monitor conformational change with infrared spectroscopy paves the way for a sensor to screen for new anticancer therapeutic agents.


Obstructing leiomyoma of the common hepatic duct bifurcation simulating a Klatskin tumor.
The first reported case of a leiomyoma of the proximal bile duct simulating a Klatskin tumor and causing obstructive cholestasis is presented. Diagnosis was made at the time of exploratory surgery. Surgical management included complete excision with Roux-en-Y hepatic-ojejunostomy.


A multicolour flow cytometry identifying defined leukocyte subsets of rainbow trout (Oncorhynchus mykiss).
The investigation of the cellular immune response in fish species has been for a long time hampered by absence of appropriate monoclonal antibodies (MAbs) recognising subset specific surface markers. Consequently, the majority of immunological studies still focus on the changes in total leukocyte numbers or describe gene pattern in lymphoid organs without any information about their cellular composition. Flow cytometric techniques are routinely used for the evaluation of the leukocyte composition in numerous vertebrate species and contributed significantly to the current knowledge of immune system. In rainbow trout is so far only a limited number of MAbs against characterised (IgM and IgT, CD8α) or unknown lineage markers on thrombocytes, myeloid cells or T cells available. By combination of several MAbs, we developed a rapid, simple, accurate and high throughput method for reliable discrimination of major leukocyte subpopulations from 10 μl of peripheral blood. Additionally, by a consecutive gating, this mixture enables the evaluation of the proportion between CD8α(+) and CD8α(-) population and provides for the first time valuable information about the kinetic of CD4(+) cells in rainbow trout. Furthermore, the combination of all antibodies within one sample reduced the hands-on time down to 90 min allowing fast and accurate estimation of cell kinetics in a high number of individuals. Thus presented findings enable the precise evaluation of the cellular components of immune system during both pathological and physiological responses and have therefore an immense potential for future applications in the development of vaccines and better understanding of fish immune system.


Polo-like kinase-1 is required for bipolar spindle formation but is dispensable for anaphase promoting complex/Cdc20 activation and initiation of cytokinesis.
Polo-like kinase-1 (Plk1) performs multiple essential functions during the cell cycle. Here we show that human Plk1-deficient cells are unable to separate their centrosomes, fail to form a bipolar spindle, and undergo a Mad2/BubR1-dependent prometaphase arrest. However, electron microscopy demonstrates that kinetochore-microtubule interactions can be established in cells lacking Plk1. In addition, co-depletion of Plk1 and survivin allows mitotic exit. This indicates that Plk1 depletion does not prevent microtubule attachment, but specifically interferes with the generation of tension, as a consequence of a failure to form a bipolar spindle. Moreover, we find that after silencing of the spindle assembly checkpoint, degradation of cyclin B1 is unaffected in cells lacking Plk1. These data indicate that activation of the anaphase promoting complex or cyclosome (APC/C)-Cdc20 complex that is under control of the spindle assembly checkpoint does not require Plk1 activity. Finally, we find that translocation of chromosome passengers and initiation of cleavage furrow ingression is unaffected in cells depleted of Plk1. Thus, our data confirm an important role of Plk1 in bipolar spindle formation, and also demonstrate that Plk1 is dispensable for APC/C-Cdc20 activation and the initiation of cytokinesis.


Postoperative cognitive dysfunction versus complaints: a discrepancy in long-term findings.
This review describes the discrepancy in findings between postoperative cognitive performance and postoperative cognitive complaints long time after an operation under general anesthesia. Shortly (from 6 hr to 1 week) after an operation a decline in cognitive performance is reported in most studies. However, long time (from 3 weeks to 1-2 years) after an operation this is rarely found although some patients are still reporting cognitive complaints. In general this kind of research is suffering from severe methodological problems (use of insensitive tests, lack of control groups, lack of parallel tests, different definitions of cognitive decline). However, these problems cannot totally explain the discrepancy in findings in the long term. Thus, there are patients who have persistent cognitive complaints long time after an operation, that cannot be measured with cognitive tests. More psychological factors such as fixation on short-term cognitive dysfunction, mood, coping style, and personality are possible explanations for these cognitive complaints in the long term. As a consequence, these factors should be a topic in future research elucidating the persistence of these cognitive complaints long time after an operation under general anesthesia.


[Agreement between measured and calculated by predictive formulas resting energy expenditure in severe and morbid obese women].
To compare measured resting energy expenditure (REE) with that predicted by formulas derived from populations with normal weight or obesity and from women with severe and morbid obesity. 66 women (aged 35.6 +/- 10.3 y and BMI of 44.7 +/- 4.9 kg/m2) were evaluated by indirect calorimetry with a metabolic monitor Deltatrac (Datex Inst., Finland), before undergoing gastric bypass. REE was calculated with the following equations: Harris-Benedict's with both actual and adjusted weight, Ireton-Jones', Mifflin's, and Carrasco's Fast Estimation, which corresponds to 16.2 kcal x kg actual weight. (mean +/- sd). Measured REE was 1797 +/- 239 kcal/day. All formulas, except Harris-Benedict's with adjusted weight, overestimated REE. The Ireton-Jones' equation presented the greater overestimation (689 +/- 329 kcal/day), whereas Mifflin's equation overestimated REE only by 6 +/- 202 kcal/day. No significant differences were detected between measured and calculated REE by Mifflin's and Carrasco's Fast Estimation. Accuracy (defined as difference between calculated and measured REE within +/- 10%) was greater with Mifflin's equation (68%), followed by Harris-Benedict's with actual weight (64%) and Carrasco's Fast Estimation (61%). By using the Bland-Altman analysis, significant correlations were observed between calculated-measured REE and mean REE (calculated + measured/2) with all equations except Carrasco's Fast Estimation. This means that all but one formula underestimate or overestimate REE depending on the level of measured REE. In severe and morbid obese women, Mifflin's and Carrasco's Fast Estimation equations provided the best performance to estimate REE. Before recommending an equation in an a subset of individuals it is necessary to make previous validation studies to determine that equation with the best predictive power for this particular group of patients.


Severe pertussis infection in infants less than 6 months of age: Clinical manifestations and molecular characterization.
We conducted a study to determine the main traits of pertussis among unimmunized infants less than 6 months of age. From August 2012 to March 2015, 141 nasopharyngeal aspirates (NPAs) were collected from infants with respiratory symptoms attending 2 major hospitals in Rome. Clinical data were recorded and analyzed. Lab-confirmation was performed by culture and realtime PCR. B. pertussis virulence-associated genes (ptxP, ptxA and prn), together with multilocus variable-number tandem repeat analysis (MLVA), were also investigated by the sequence-based analysis on the DNAs extracted from positive samples. Antibiotic susceptibility with Etest was defined on 18 viable B. pertussis isolates. Samples from 73 infants resulted positives for B. pertussis. The median age of the patients was 45 d (range 7-165); 21 infants were treated with macrolides before hospital admission. Cough was reported for a median of 10 d before admission and 18 d after hospital discharge among infected infants, 84% of whom showed paroxysmal cough. No resistance to macrolides was detected. Molecular analysis identified MT27 as the predominant MLVA profile, combined with ptxP3-ptxA1-prn2 associated virulence genes. Although our data may not be generalized to the whole country, they provide evidence of disease severity among infants not vaccinated against pertussis. Moreover, genetically related B. pertussis strains, comprising allelic variants of virulence associated genes, were identified.


Grandparenthood and risk of mortality: Findings from the Health and Retirement Study.
Grandparenthood constitutes a significant role for older adults and may have important health implications. Our study examines the grandparenthood-mortality nexus, controlling for an array of potentially confounding variables. Longitudinal survey data from the Health and Retirement Study (HRS) were used, comprising twelve biennial waves from 1992 to 2014 with linked data on vital status derived from the National Death Index. The sample included 27,463 participants aged ≥51 years with at least one child. Cox proportional hazard models tested the association between grandparenthood and mortality risk with adjustment for socio-demographic variables, for social variables including characteristics of and contact with children, and for health variables, including measures of general, functional and mental health. Grandparenthood overall was unassociated with mortality risk in both women and men. However, the subpopulation of younger, partnered grandmothers with a larger number of grandchildren tended to exhibit a substantial increase in mortality risk as compared to women without grandchildren.


FSH inhibits AMH to support ovarian estradiol synthesis in infantile mice.
Anti-Müllerian hormone (AMH) regulates ovarian function in cyclic females, notably by preventing premature follicle-stimulating hormone (FSH)-mediated follicular growth and steroidogenesis. Its expression in growing follicles is controlled by FSH and by estradiol (E2). In infantile females, there is a transient increase in the activity of the gonadotrope axis, as reflected by elevated levels of both gonadotropins and E2. We previously demonstrated in mice that elevated FSH concentrations are necessary to induce E2 production by preantral/early antral follicles through the stimulation of aromatase expression without supporting their growth. However, whether this action of FSH could involve AMH is unknown. Here, we show that Amh mRNA and protein abundance and serum AMH levels are elevated in infantile mouse females, compared with those in adults. By experimentally manipulating FSH and E2 levels in infantile mice, we demonstrate that high FSH concentrations lower Amh expression specifically in preantral/early antral follicles, whereas E2 has no effect. Importantly, treatment of infantile ovaries in organotypic cultures with AMH decreases FSH-mediated expression of Cyp19a1 aromatase, but it does not alter the expression of cyclin D2-mediating granulosa cell proliferation. Overall, our data indicate that the infantile elevation in FSH levels suppresses Amh expression in preantral/early antral follicles, thereby favoring Cyp19a1 aromatase expression and E2 production. Together with recent discoveries that AMH can act on both the hypothalamus and the pituitary to increase gonadotropin levels, this work suggests that AMH is a critical regulator of the gonadotrope axis during the infantile period, thereby contributing to adult reproductive function programming.


Mycobacterium leprae is identified in the oral mucosa from paucibacillary and multibacillary leprosy patients.
In leprosy, the nasal mucosa is considered as the principal route of transmission for the bacillus Mycobacterium leprae. The objective of this study was to identify M. leprae in the oral mucosa of 50 untreated leprosy patients, including 21 paucibacillary (PB) and 29 multibacillary (MB) patients, using immunohistochemistry (IHC), with antibodies against bacillus Calmette-Guérin (BCG) and phenolic glycolipid antigen-1 (PGL-1), and polymerase chain reaction (PCR), with MntH-specific primers for M. leprae, and to compare the results. The material was represented by 163 paraffin blocks containing biopsy samples obtained from clinically normal sites (including the tongue, buccal mucosa and soft palate) and visible lesions anywhere in the oral mucosa. All patients and 158 available samples were included for IHC study. Among the 161 available samples for PCR, 110 had viable DNA. There was viable DNA in at least one area of the oral mucosa for 47 patients. M. leprae was detected in 70% and 78% of patients using IHC and PCR, respectively, and in 94% of the patients by at least one of the two diagnostic methods. There were no differences in detection of M. leprae between MB and PB patients. Similar results were obtained using anti-BCG and anti-PGL-1 antibodies, and immunoreactivity occurred predominantly on free-living bacteria on the epithelial surface, with a predilection for the tongue. Conversely, there was no area of predilection according to the PCR results. M. leprae is present in the oral mucosa at a high frequency, implicating this site as a potential means of leprosy transmission.


Automated Mobile Phone-Based Mental Health Resource for Homeless Youth: Pilot Study Assessing Feasibility and Acceptability.
Youth experiencing housing instability have higher rates of mental health problems than their housed peers. Few studies have evaluated technological resources for homeless youth to determine how to effectively engage and reach them. The primary aims of this pilot study were to establish the feasibility (as measured by phone retention rates) and acceptability (ie, participant ratings of resources) of delivering automated mental health resources via smartphone technology. Youth aged 16 to 25 years (N=100) were recruited through homeless shelter agencies in the Chicago metropolitan area. Eligible participants completed a baseline assessment and received a smartphone with a 3-month data plan. The phone was preloaded with several apps designed to promote mental health wellness and provide real-time resources. One app specifically designed for this study, Pocket Helper 2.0, sent participants daily surveys and tips via push notification. The tips focused on coping and motivation, and the surveys assessed mood. This app also included an automated self-help system with brief cognitive behavioral interventions (5-10 min) and access to several interactive mobile tools, including a crisis text line, a telephone hotline, a crowd-based emotional support tool, and an app providing up-to-date information on social service and mental health resources for homeless youth in Chicago. Participants completed assessments at 3 and 6 months. Some individuals (23%, 23/100) experienced problems with the phones (eg, theft, loss, and technological issues) throughout the study. Participant retention at the midpoint was moderate, with 48% (48/100) of youth responding to the 3-month surveys. At 6 months, only 19% (19/100) of the total sample responded to the end point survey. Overall, 63% (30/48) to 68% (13/19) of respondents at both time points reported benefiting from the intervention; however, participant usage and satisfaction varied with the different features. At both time points, participants reported receiving the most benefit from the daily tips and daily surveys. Daily tips that were most preferred by participants involved motivational tips related to overcoming struggles and making progress in life. Aside from the tips and surveys, the most used features were the app providing up-to-date resources and the automated self-help system. Interactive features, including the telephone hotline and crowd-based emotional support tool, were the least used features and were rated as the least beneficial. Automated mental health interventions seem to be an acceptable way to engage homeless youth in mental health support. The participants preferred fully automated features and brief interventions over features requiring interaction with others or more engagement. Future research should explore ways to retain homeless youth in interventions and evaluate the clinical impact of automated technology-based interventions for improving mental health. ClinicalTrials.gov NCT03776422; https://clinicaltrials.gov/ct2/show/NCT03776422.


Novel H-Bonded Synthons in Copper Supramolecular Frameworks with Aminoethylpiperazine-Based Ligands. Synthesis, Structure and Catalytic Activity.
New Schiff base complexes [Cu2(HL1)(L1)(N3)3]∙2H2O (1) and [Cu2L2(N3)2]∙H2O (2) were synthesized. The crystal structures of 1 and 2 were determined by single-crystal X-ray diffraction analysis. The HL1 ligand results from the condensation of salicylaldehyde and 1-(2-aminoethyl)piperazine, while a new organic ligand, H2L2, was formed by the dimerization of HL1 via a coupling of two piperazine rings of HL1 on a carbon atom coming from DMF solvent. The dinuclear building units in 1 and 2 are linked into complex supramolecular networks through hydrogen and coordination bondings, resulting in 2D and 1D architectures, respectively. Single-point and broken-symmetry DFT calculations disclosed negligible singlet-triplet splittings within the dinuclear copper fragments in 1 and 2. Catalytic studies showed a remarkable activity of 1 and 2 towards cyclohexane oxidation with H2O2 in the presence of nitric acid and pyridine as promoters and under mild conditions (yield of products up to 21%). Coordination compound 1 also acts as an active catalyst in the intermolecular coupling of cyclohexane with benzamide using di-tert-butyl peroxide (tBuOOtBu) as a terminal oxidant. Conversion of benzamide at 55% was observed after 24 h reaction time. By-product patterns and plausible reaction mechanisms are discussed.