Prognostic signature associated with radioresistance in head and neck cancer via transcriptomic and bioinformatic analyses.
Radiotherapy is an indispensable treatment modality in head and neck cancer (HNC), while radioresistance is the major cause of treatment failure. The aim of this study is to identify a prognostic molecular signature associated with radio-resistance in HNC for further clinical applications. Affymetrix cDNA microarrays were used to globally survey different transcriptomes between HNC cell lines and isogenic radioresistant sublines. The KEGG and Partek bioinformatic analytical methods were used to assess functional pathways associated with radioresistance. The SurvExpress web tool was applied to study the clinical association between gene expression profiles and patient survival using The Cancer Genome Atlas (TCGA)-head and neck squamous cell carcinoma (HNSCC) dataset (n = 283). The Kaplan-Meier survival analyses were further validated after retrieving clinical data from the TCGA-HNSCC dataset (n = 502) via the Genomic Data Commons (GDC)-Data-Portal of National Cancer Institute. A panel maker molecule was generated to assess the efficacy of prognostic prediction for radiotherapy in HNC patients. In total, the expression of 255 molecules was found to be significantly altered in the radioresistant cell sublines, with 155 molecules up-regulated 100 down-regulated. Four core functional pathways were identified to enrich the up-regulated genes and were significantly associated with a worse prognosis in HNC patients, as the modulation of cellular focal adhesion, the PI3K-Akt signaling pathway, the HIF-1 signaling pathway, and the regulation of stem cell pluripotency. Total of 16 up-regulated genes in the 4 core pathways were defined, and 11 over-expressed molecules showed correlated with poor survival (TCGA-HNSCC dataset, n = 283). Among these, 4 molecules were independently validated as key molecules associated with poor survival in HNC patients receiving radiotherapy (TCGA-HNSCC dataset, n = 502), as IGF1R (p = 0.0454, HR = 1.43), LAMC2 (p = 0.0235, HR = 1.50), ITGB1 (p = 0.0336, HR = 1.46), and IL-6 (p = 0.0033, HR = 1.68). Furthermore, the combined use of these 4 markers product an excellent result to predict worse radiotherapeutic outcome in HNC (p < 0.0001, HR = 2.44). Four core functional pathways and 4 key molecular markers significantly contributed to radioresistance in HNC. These molecular signatures may be used as a predictive biomarker panel, which can be further applied in personalized radiotherapy or as radio-sensitizing targets to treat refractory HNC.


Virgin Camellia Seed Oil Improves Glycolipid Metabolism in the Kidney of High Fat-Fed Rats through AMPK-SREBP Pathway.
Camellia seed oil (CO) is used as edible oil in southern China because of its excellent fatty acid composition and abundant bioactive compounds. Chronic kidney disease (CKD) is one of the most common chronic degenerative diseases in China, and active compounds in vegetable oil, like virgin olive oil, have been demonstrated to be efficacious in the management of CKD. In this study, virgin CO was refined using a standard process. The refining had minimal impact on the fatty acid composition, but significantly reduced the presence of bioactive compounds like polyphenols in CO. Sprague-Dawley (SD) rats fed with high fat diet (Group G) were treated with either virgin (Group Z) or refined CO (Group R). The oral administration of CO alleviated lipid accumulation and decreased body and kidney weight gain. Furthermore, treatment with virgin CO increased the renal ATP content. The renal expression levels of AMPK and key enzymes involved in fatty acid oxidation (CPT-1 and ACOX1) and glycolysis (HK, PFK, PK and GAPDH) were up-regulated in Group Z, thereby enhancing the ATP production. Virgin CO treatment downregulated the expression level of SREBP2 and its downstream target genes, such as ACC, FAS, and HMGCR, which reduced lipid synthesis. These findings indicate that virgin CO improves glycolipid metabolism and restores energy homeostasis in the kidneys of rats fed with a high-fat diet by modulating the AMPK-SREBP-signaling pathway, suggesting the potential of active compounds in virgin CO for managing the renal failure associated with glycolipid dysmetabolism.


Hamstring graft size prediction: a prospective clinical evaluation.
Recently we retrospectively collected clinical data to predict hamstring graft diameter. Prospective data collection will improve and further define prediction of hamstring graft size. Clinical anthropometric data can be used to predict hamstring graft size. Cohort study (prevalence); Level of evidence, 1. Fifty consecutive patients with anterior cruciate ligament deficiency scheduled for reconstruction using hamstring autograft were prospectively evaluated. Preoperatively we recorded height, weight, body mass index, age, gender, leg length, thigh length, shank length, bilateral thigh circumference, and Tegner score. Intraoperative measurements of both the gracilis and semitendinosus tendons were made, including absolute length before fashioning the graft and final diameter of the quadrupled graft using sizing tubes calibrated to 0.5 mm. Bivariate correlation coefficients (Pearson r) were calculated to identify relationships among clinical data and intraoperatively measured hamstring graft length and diameter. Strongest correlations for graft lengths were height and leg length measurements. Shorter persons with shorter leg, thigh, and shank lengths tended to have shorter gracilis and semitendinosus grafts. Likewise, the strongest correlations for graft diameter were weight and thigh circumference. Self-reported activity level and age did not correlate. Gender comparison revealed that women who were shorter, lighter, and had smaller body mass indices were more likely to have smaller graft diameters and shorter graft lengths. Patients weighing less than 50 kg, less than 140 cm in height, with less than 37 cm thigh circumference, and with body mass index less than 18 should be considered at high risk for having a quadrupled hamstring graft diameter less than 7 mm. When separated by gender, small graft diameters are most likely in older, short, female subjects with small thigh circumferences or young, skinny, male subjects with small thigh circumferences and low body mass index. Common clinical measurements can be used for preoperative identification of patients at risk for insufficient graft tissue and would be useful for patient counseling and alternative graft source planning.


Cell cycle control as a basis for cancer drug development (Review).
Normal cell cycle progression relies on the cell's ability to translate extracellular signals such as mitogenic stimuli and intact extracellular matrices in order to efficiently replicate DNA and divide. Cyclin dependent kinases (cdks) respond to these signals and are largely responsible for positively pushing cells through the cell cycle. Due to their pivotal role in cell division, nature has evolved elaborate mechanisms to regulate the kinase activity of cdks. Cyclins are cdk binding partners which are required for kinase activity and their protein levels are intimately linked to the cell cycle stage. A variety of other cdk regulators such as phosphorylation events, natural inhibitors and complex stability are discussed. Phosphorylation of various cdk substrates results in diverse outcomes such as changes in gene expression, formation of prereplicative complexes and breakdown of the nuclear envelope. Cancer cells evolve in part by over-riding normal cell cycle regulation. Abnormal cdk activity is accomplished by cyclin amplification, cdk or substrate mutation as well as inactivation of inhibitors. The selective growth advantage of cancer cells also stems from amplification of positive growth signals, mutation of checkpoint and surveillance genes as well as deregulation of programmed cell death or apoptosis. The full potential of cancer therapies, such as small molecule inhibitors and gene therapy among others, focusing on our knowledge of cell cycle regulation has yet to be reached.


Impact of Tourette Syndrome on Education.
Previous studies have shown that Tourette syndrome (TS) has an impact on academic achievements. The aim of this study was to investigate the association between the severity of tics and comorbidities and educational outcomes. From 2005 to 2007, 395 participants were included in a large cohort (314 with TS and 81 controls) and the mean age was 12.60 ± 2.64 years. The cohort was re-examined after 4 to 8 years (median 5.6) where n = 276 participants (223 with TS and 53 controls) were included with a mean age of 18.52 ± 2.73 years. At both time points, severity of tics and the presence and severity of psychiatric comorbidity were assessed. Educational achievements were assessed through structured interviews. Children with TS had a lower passing rate at lower secondary and high school compared to healthy controls. More severe vocal tics were associated with fewer passing lower secondary school at a prospective level. At a cross-sectional level, more severe motor tics were associated with fewer passing high school. Tic severity only influenced children with TS without comorbidity. The severity of comorbidity was found to be associated with the educational level at a longitudinal view, but not cross-sectional. Overall, children with TS had a lower passing rate at lower secondary school and high school compared to healthy controls. We found that this difference was more likely driven by the severity of comorbidities than tic severity. It is important to be aware of academic achievement in children with TS in order to give them the right support and thereby optimize educational opportunities.


Heavy carbon nanodots 2: plasmon amplification in Quanta Plate™ wells and the correlation with the synchronous scattering spectrum.
Brominated carbon nanodots are a new carbon nanostructure that exhibits strong phosphorescence without fixation. Herein we report plasmonic amplification of this phosphorescence in silver-coated Quanta Plate™ wells, a technique called metal-enhanced phosphorescence (MEP). Subsequently we correlate the excitation and emission components of brominated carbon nanodots to their respective enhancement values. These properties are then discussed in relation to the synchronous scattering spectrum of the plasmonic substrate, in the first report of its kind for MEP. These results set the foundation for expanded application of carbon nanodots, as the photophysical characteristics of phosphorescence are improved, and augment the growing understanding of MEP.


Resistant "candidal intertrigo": could inverse psoriasis be the true culprit?
Inverse psoriasis is a disorder of intertriginous areas of the skin that can easily masquerade as candidal intertrigo. Candidal rashes are commonly encountered in primary care and typically respond promptly to therapy. When treatment fails, nonadherence to treatment and medication resistance often are suspected; however, the possibility of an incorrect diagnosis should also be entertained. This article presents the case of a patient with inverse psoriasis who was misdiagnosed with recurrent candidal intertrigo multiple times. The diagnosis and treatment of inverse psoriasis is reviewed, and other conditions that may be confused with Candida and inverse psoriasis, including bacterial intertrigo, tinea, and seborrheic dermatitis, are discussed. When confronted with a case of "resistant Candida," consideration of inverse psoriasis and other Candida mimics can allow physicians to diagnose and treat these conditions more effectively, avoiding the frustration experienced by our patient.


A novel percutaneous system for bone graft delivery and containment for elevation and stabilization of vertebral compression fractures. Technical note.
Vertebroplasty and kyphoplasty are minimally invasive procedures used to treat persistently symptomatic vertebral compression fractures (VCFs). Both interventions usually involve injection of polymethyl methacrylate (PMMA). The purpose of this technical note was to review the theory and surgical technique for a novel percutaneous system for fracture reduction and stabilization of VCFs by using bone graft. This technical note highlights the Optimesh system as an alternative method of minimally invasive VCF reduction and stabilization with the delivery of a bone graft containment device. Instead of using PMMA as in vertebroplasty or kyphoplasty, this system allows the delivery of allograft and/or autograft bone, with its osteoinductive, osteoconductive, and osteogenic properties. This system allows for restoration of sagittal alignment of the spine with direct control of bone graft delivery by using a mesh graft containment device that allows for ingrowth of new bone and vascular tissue.


Partial Tmem106b reduction does not correct abnormalities due to progranulin haploinsufficiency.
Loss of function mutations in progranulin (GRN) are a major cause of frontotemporal dementia (FTD). Progranulin is a secreted glycoprotein that localizes to lysosomes and is critical for proper lysosomal function. Heterozygous GRN mutation carriers develop FTD with TDP-43 pathology and exhibit signs of lysosomal dysfunction in the brain, with increased levels of lysosomal proteins and lipofuscin accumulation. Homozygous GRN mutation carriers develop neuronal ceroid lipofuscinosis (NCL), an earlier-onset lysosomal storage disorder caused by severe lysosomal dysfunction. Multiple genome-wide association studies have shown that risk of FTD in GRN mutation carriers is modified by polymorphisms in TMEM106B, which encodes a lysosomal membrane protein. Risk alleles of TMEM106B may increase TMEM106B levels through a variety of mechanisms. Brains from FTD patients with GRN mutations exhibit increased TMEM106B expression, and protective TMEM106B polymorphisms are associated with decreased TMEM106B expression. Together, these data raise the possibility that reduction of TMEM106B levels may protect against the pathogenic effects of progranulin haploinsufficiency. We crossed Tmem106b +/- mice with Grn +/- mice, which model the progranulin haploinsufficiency of GRN mutation carriers and develop age-dependent social deficits and lysosomal abnormalities in the brain. We tested whether partial Tmem106b reduction could normalize the social deficits and lysosomal abnormalities of Grn +/- mice. Partial reduction of Tmem106b levels did not correct the social deficits of Grn +/- mice. Tmem106b reduction also failed to normalize most lysosomal abnormalities of Grn +/- mice, except for β-glucuronidase activity, which was suppressed by Tmem106b reduction and increased by progranulin insufficiency. These data do not support the hypothesis that Tmem106b reduction protects against the pathogenic effects of progranulin haploinsufficiency, but do show that Tmem106b reduction normalizes some lysosomal phenotypes in Grn +/- mice.


Oxidative stress in atherosclerosis-prone mouse is due to low antioxidant capacity of mitochondria.
Atherosclerotic disease remains a leading cause of death in westernized societies, and reactive oxygen species (ROS) play a pivotal role in atherogenesis. Mitochondria are the main intracellular sites of ROS generation and are also targets for oxidative damage. Here, we show that mitochondria from atherosclerosis-prone, hypercholesterolemic low-density lipoprotein (LDL) receptor knockout mice have oxidative phosphorylation efficiency similar to that from control mice but have a higher net production of ROS and susceptibility to develop membrane permeability transition. Increased ROS production was observed in mitochondria isolated from several tissues, including liver, heart, and brain, and in intact mononuclear cells from spleen. In contrast to control mitochondria, knockout mouse mitochondria did not sustain a reduced state of matrix NADPH, the main source of antioxidant defense against ROS. Experiments in vivo showed faster liver secretion rates and de novo synthesis of triglycerides and cholesterol in knockout than in control mice, suggesting that increased lipogenesis depleted the reducing equivalents from NADPH and generated a state of oxidative stress in hypercholesterolemic knockout mice. These data provide the first evidence of how oxidative stress is generated in LDL receptor defective cells and could explain the increased LDL oxidation, cell death, and atherogenesis seen in familiar hypercholesterolemia.


A theory-based multi-component intervention to increase reactive balance measurement by physiotherapists in three rehabilitation hospitals: an uncontrolled single group study.
Most implementation interventions in rehabilitation, including physiotherapy, have used passive, non-theoretical approaches without demonstrated effectiveness. The goal of this study was to improve an important domain of physiotherapy practice - reactive balance measurement - with a targeted theory-based multi-component intervention developed using the Theoretical Domains Framework. The primary objective was to determine documented reactive balance measure use in a 12-month baseline, during, and for three months post- intervention. An uncontrolled before-and-after study was completed with physiotherapists at three urban adult rehabilitation hospitals in Ontario, Canada. The 12-month intervention included group meetings, local champions, and health record modifications for a validated reactive balance measure. The primary outcome was the proportion of records with a documented reactive balance measure when balance was assessed pre-, during- and post-intervention. Secondary outcomes were changes in use, knowledge, and confidence post-intervention, differences across sites, and intervention satisfaction. Reactive balance was not measured in any of 211 eligible pre-intervention records. Thirty-three physiotherapists enrolled and 28 completed the study. Reactive balance was measured in 31% of 300 eligible records during-intervention, and in 19% of 90 eligible records post-intervention (p < 0.04). Knowledge and confidence significantly increased post-intervention (all p < 0.05). There were significant site differences in use during- and post-intervention (all p < 0.05). Most participants reported satisfaction with intervention content (71%) and delivery (68%). Reactive balance measurement was greater among participants during-intervention relative to the baseline, and use was partially sustained post-intervention. Continued study of intervention influences on clinical reasoning and exploration of site differences is warranted.


[Akatinol memantin in Alzheimer's disease: clinico-immunological correlates].
16 patients with Alzheimer's disease were treated with akatinol memantine. The levels of production of interleukin-1 (IL-1) in lymphocyte culture were evaluated both before and after the treatment that lasted for 2-4.5 months. Before the therapy a significant (p < 0.05) increase in the production of IL-1 was observed in patients with mild and moderate dementia, with early onset of the disease and with the duration of the disease up to 9 years as compared with a control group (24 healthy individuals). After the treatment the production of IL-1 was decreased in patients with moderate dementia, with early onset, and with hereditary form of the disease. Decrease of the level of IL-1 production after the treatment corresponded to improvement of the cognitive functions of the patient. In cases of unsatisfactory treatment (without any improvement of a clinical state) IL-1 production was at higher levels (p < 0.05) as compared with the control values. It is suggested that initial high level of IL-1 production could serve as a predictor of efficiency of therapy with akatinol memantine in patients with early onset of Alzheimer's disease.


The MetabolomeExpress Project: enabling web-based processing, analysis and transparent dissemination of GC/MS metabolomics datasets.
Standardization of analytical approaches and reporting methods via community-wide collaboration can work synergistically with web-tool development to result in rapid community-driven expansion of online data repositories suitable for data mining and meta-analysis. In metabolomics, the inter-laboratory reproducibility of gas-chromatography/mass-spectrometry (GC/MS) makes it an obvious target for such development. While a number of web-tools offer access to datasets and/or tools for raw data processing and statistical analysis, none of these systems are currently set up to act as a public repository by easily accepting, processing and presenting publicly submitted GC/MS metabolomics datasets for public re-analysis. Here, we present MetabolomeExpress, a new File Transfer Protocol (FTP) server and web-tool for the online storage, processing, visualisation and statistical re-analysis of publicly submitted GC/MS metabolomics datasets. Users may search a quality-controlled database of metabolite response statistics from publicly submitted datasets by a number of parameters (eg. metabolite, species, organ/biofluid etc.). Users may also perform meta-analysis comparisons of multiple independent experiments or re-analyse public primary datasets via user-friendly tools for t-test, principal components analysis, hierarchical cluster analysis and correlation analysis. They may interact with chromatograms, mass spectra and peak detection results via an integrated raw data viewer. Researchers who register for a free account may upload (via FTP) their own data to the server for online processing via a novel raw data processing pipeline. MetabolomeExpress https://www.metabolome-express.org provides a new opportunity for the general metabolomics community to transparently present online the raw and processed GC/MS data underlying their metabolomics publications. Transparent sharing of these data will allow researchers to assess data quality and draw their own insights from published metabolomics datasets.


Variation in Drosophila sensory bristle number at 'Evolution Canyon'.
'Evolution Canyon' on Mount Carmel, Israel, displays highly contrasting physical and biotic environments on a micro-geographic scale, and is a natural laboratory for investigating genetic responses to variable and extreme environments across species. Samples of Drosophila melanogaster and D. simulans were collected from three sites each on the north- and south-facing slopes of the canyon along altitudinal transects, and one site on the valley floor. Numbers of abdominal and sternopleural sensory bristles were recorded for each of these subpopulations in three thermal environments. In D. simulans, sternopleural bristle number exhibited micro-geographic differentiation between the north- and south-facing slopes, while abdominal bristle number was stable across subpopulations. In D. melanogaster, the magnitudes of the difference in mean sternopleural bristle number between the north- and south-facing slopes and of mean abdominal bristle number along the altitudinal gradients were both conditional on rearing temperature. Thus, the pattern of genetic variation between sites was consistent with underlying heterogeneity of genetic mechanisms for response to the same environmental gradients between traits and sibling species. In contrast, the genetic architecture of bristle number at the level of variation within populations was very similar between species for the same bristle trait, although the two traits differed in the relative contribution of genotype by temperature and genotype by sex interaction.


Spectroscopic determination of acid dissociation constants of N-substituted-6-acylbenzothiazolone derivatives.
The acid dissociation constants of twelve novel drug precursor N-substituted-6-acylbenzothiazolone derivatives were determined by using the UV-vis spectroscopic technique. The protonation and deprotonation behaviors of the investigated molecules were researched from the super basic to super acid regions (i.e., 8 mol·L(-1) KOH to 98% H(2)SO(4)) including the pH region. It is observed that all of the molecules are protonated in the super acidic region. The calculated relative stability values of possible tautomer structures indicate that the keto form of investigated molecules is favored over the enol form. It was predicted that protonation occurs at the amide (oxo) group found in the keto form.


The impact of neighborhood quality, perceived stress, and social support on depressive symptoms during pregnancy in African American women.
Living in a lower-quality neighborhood is associated with higher levels of depressive symptoms in the general population as well as among pregnant and postpartum women. However, little is known of the important pathways by which this association occurs. We proposed a model in which perceived stress and social support mediated the effects of neighborhood quality on depressive symptoms during pregnancy (measured by the 20-item Center for Epidemiologic Studies-Depression, CES-D, scale) in a sample of 1383 African American women from the Detroit metropolitan area interviewed during their delivery hospitalization. Using structural equation modeling (SEM), we built a latent variable of neighborhood quality using 4 measures (neighborhood disorder, neighborhood safety/danger, walking environment, overall rating). We then tested two SEM mediation models. We found that lower neighborhood quality was associated with higher prevalence of depressive symptoms during pregnancy (standardized total effect = .16, p = .011). We found that perceived stress partially mediated the neighborhood quality association with depressive symptoms. Although the association of social support with depressive symptoms was negligible, social support mediated associations of neighborhood quality with perceived stress [standardized path coefficient = .38 (.02), p = .009]. Our results point to the need for public health, health care, as well as non-health related interventions (e.g. crime prevention programs) to decrease overall exposure to stressors, as well as stress levels of women living in poor quality neighborhoods. Interventions that increase the levels of social support of women during pregnancy are also needed for their potential to decrease stress and ultimately improve mental health at this important time in the life course.


Perioperative chemotherapy with bevacizumab and liver resection for colorectal cancer liver metastasis.
Surgery remains the only curative option for patients with colorectal cancer liver metastases (CRLM). Perioperative chemotherapeutic strategies have become increasingly popular in the treatment of CRLM. Although the role of bevacizumab (Bev) in this setting remains unclear, its widespread use has raised concerns about the use of Bev as part of perioperative chemotherapy. We retrospectively reviewed all patients who received Bev and underwent liver resection between July 2004 and July 2008 at the McGill University Health Center. Chemotherapy-related toxicity, response to chemotherapy, surgical morbidity and mortality, liver function and survival data were assessed. A total of 35 patients were identified. Of these, 26 (74.3%) patients received oxaliplatin-based cytotoxic chemotherapy, six (17.1%) received irinotecan-based therapy and the remainder received both agents. A total of 17 patients (48.6%) underwent portal vein embolization prior to resection and 12 (34.3%) underwent staged resection for extensive bilobar disease. A median of six cycles of preoperative Bev were administered. Nine patients (25.7%) experienced grade 3 or higher chemotherapy-related toxicities. Four events were deemed to be related to Bev. The overall response rate was 65.7% (complete and partial response). One patient progressed on therapy, but this did not prevent R0 resection. The incidence of postoperative morbidity was 42.3%. A total of 21.7% of complications were Clavien grade 3 or higher. There were no perioperative mortalities. There were no cases of severe sinusoidal injury or steatohepatitis. The Kaplan-Meier estimate of 4-year survival was 52.5%. These data confirm the safety of chemotherapy regimens which include Bev in the perioperative setting and demonstrate that such perioperative chemotherapy in patients with CRLM does not adversely affect patient outcome. There was no increase in perioperative morbidity compared with published rates. The addition of Bev to standard chemotherapy may improve response rates, which may, in turn, impact favourably on patient survival.


The contribution of linear perspective cues and texture gradients in the perceptual rescaling of stimuli inside a Ponzo illusion corridor.
We examined the influence of linear perspective cues and texture gradients in the perceptual rescaling of stimuli over a highly-salient Ponzo illusion of a corridor. We performed two experiments using the Method of Constant Stimuli where participants judged the size of one of two rings. In experiment 1, one ring was presented in the upper visual-field at the end of the corridor and the other in the lower visual-field at the front of the corridor. The perceived size of the top and bottom rings changed as a function of the availability of linear perspective and textures. In experiment 2, only one ring was presented either at the top or the bottom of the image. The perceived size of the top but not the bottom ring changed as a function of the availability of linear perspective and textures. In both experiments, the effects of the cues were additive. Perceptual rescaling was also stronger for the top compared to the bottom ring. Additional eye-tracking revealed that participants tended to gaze more in the upper than the lower visual-field. These findings indicate that top-down mechanisms provide an important contribution to the Ponzo illusion. Nonetheless, additional maximum likelihood estimation analyses revealed that linear perspective fulfilled a greater contribution in experiment 2, which is suggestive of a bottom-up mechanism. We conclude that both top-down and bottom-up mechanisms play important roles. However, the former seems to fulfil a more prominent role when both stimuli are presented in the image.


New generation of nitric oxide-releasing porous materials: Assessment of their potential to regulate biological functions.
Nitric oxide (NO) presents innumerable biological roles, and its exogenous supplementation for therapeutic purposes has become a necessity. Some nanoporous materials proved to be potential vehicles for NO with high storage capacity. However, there is still a lack of information about their efficiency to release controlled NO and if they are biocompatible and biologically stable. In this work, we address this knowledge gap starting by evaluating the NO release and stability under biological conditions and their toxicity with primary keratinocyte cells. Titanosilicates (ETS-4 and ETS-10 types) and clay-based materials were the materials under study, which have shown in previous studies suitable NO gas adsorption/release rates. ETS-4 proved to be the most promising material, combining good biocompatibility at 180 μg/mL, stability and slower NO release. ETS-10 and ETAS-10 showed the best biocompatibility at the same concentration and, in the case of clay-based materials, CoOS is the least toxic of those tested and the one that releases the highest NO amount. The potentiality of these new NO donors to regulate biological functions was assessed next by controlling the mitochondrial respiration and the cell migration. NO-loaded ETS-4 regulates O2 consumption and cell migration in a dose-dependent manner. For cell migration, a biphasic effect was observed in a narrow range of ETS-4 concentration, with a stimulatory effect becoming inhibitory just by doubling ETS-4 concentration. For the other materials, no effective regulation was achieved, which highlights the relevance of the new assessment presented in this work for nanoporous NO carriers that will pave the way for further developments.


Development of a database of amino acid sequences for human colon carcinoma proteins separated by two-dimensional polyacrylamide gel electrophoresis.
The tandem use of preparative two-dimensional polyacrylamide gel electrophoresis (2-DE) and electroblotting onto polyvinylidene difluoride membranes has been employed to rapidly isolate a number of proteins from a crude cell extract of a human colon carcinoma cell line (LIM 1863). The immobilized proteins were located by staining with Coomassie Brilliant Blue R-250, and selected protein spots were excised and subjected to Edman degradation. Our results demonstrate that overall sequence yields in the 3-20 pmol range can be achieved on protein spots from four identical 2-DE gels; approximately 150-200 micrograms of total protein was applied to a single 2-DE gel. An approximate two-fold increase in sensitivity of phenylthiohydantoin-amino acid detection (subpicomole range) was achieved by fitting our commercial sequencers with a simple sample transfer device which permitted the analysis of the total phenylthiohydantoin-amino acid derivative. N-Terminal amino acid sequence data was obtained for thirteen electroblotted proteins. All of these sequences positively matched those of proteins of known structure listed in the available protein sequence databases. Approximately 40% of the electroblotted proteins did not yield N-terminal sequence information, presumably because they had blocked N-termini (either naturally or artifactually). Internal amino acid sequence information was obtained from three proteins isolated by preparative 2-DE. This was achieved by in situ digestion of the proteins in the gel matrix with Staphylococcus aureus V8 protease, electrophoresis of the generated peptides in a one-dimensional gel, electrotransfer of the peptides to a polyvinylidene difluoride membrane and microsequence analysis of the electroblotted peptides.


Multiple solenoidal microcoil probes for high-sensitivity, high-throughput nuclear magnetic resonance spectroscopy.
Two designs for incorporating multiple solenoidal microcoils into a single probe head are presented to increase the throughput of high-resolution NMR. Through a combination of radio frequency switches and low-noise amplifiers, multiple NMR spectra can be acquired in the same time as a single spectrum from a conventional probe consisting of one coil. Since this method does not compromise sensitivity with regard to the individual microcoils, throughput increases linearly with the number of coils. Only one receiver is needed, and data acquisition parameters can be optimized for each sample. Specifically, a four-coil system has been implemented for proton NMR at 250 MHz using a wide-bore magnet, with an observe volume of 28 nL for each microcoil. Signal cross-contamination was approximately 0.2% between individual coils, and simultaneous one- and two-dimensional spectra have been obtained from samples of fructose, galactose, adenosine triphosphate, and chloroquine (7 nmol of each compound). A more compact two-coil configuration has also been designed for operation at 500 MHz, with observe volumes of 5 and 31 nL for the two coils. One- and two-dimensional spectra were acquired from samples of 1-butanol (55 nmol) and ethylbenzene (250 nmol).


Post-translational regulation of plant immunity.
Plants have evolved multi-layered molecular defense strategies to protect against pathogens. Plant immune signaling largely relies on post-translational modifications (PTMs) to induce rapid alterations of signaling pathways to achieve a response that is appropriate to the type of pathogen and infection pressure. In host cells, dynamic PTMs have emerged as powerful regulatory mechanisms that cells use to adjust their immune response. PTM is also a virulence strategy used by pathogens to subvert host immunity through the activities of effector proteins secreted into the host cell. Recent studies focusing on deciphering post-translational mechanisms underlying plant immunity have offered an in-depth view of how PTMs facilitate efficient immune responses and have provided a more dynamic and holistic view of plant immunity.


JNK signaling regulates E-cadherin junctions in germline cysts and determines primordial follicle formation in mice.
Physiologically, the size of the primordial follicle pool determines the reproductive lifespan of female mammals, while its establishment largely depends on a process of germline cyst breakdown during the perinatal period. The mechanisms regulating this process are poorly understood. Here we demonstrate that c-Jun amino-terminal kinase (JNK) signaling is crucial for germline cyst breakdown and primordial follicle formation. JNK was specifically localized in oocytes and its activity increased as germline cyst breakdown progressed. Importantly, disruption of JNK signaling with a specific inhibitor (SP600125) or knockdown technology (Lenti-JNK-shRNAs) resulted in significantly suppressed cyst breakdown and primordial follicle formation in cultured mouse ovaries. Our results show that E-cadherin is intensely expressed in germline cysts, and that its decline is necessary for oocyte release from the cyst. However, inhibition of JNK signaling leads to aberrantly enhanced localization of E-cadherin at oocyte-oocyte contact sites. WNT4 expression is upregulated after SP600125 treatment. Additionally, similar to the effect of SP600125 treatment, WNT4 overexpression delays cyst breakdown and is accompanied by abnormal E-cadherin expression patterns. In conclusion, our results suggest that JNK signaling, which is inversely correlated with WNT4, plays an important role in perinatal germline cyst breakdown and primordial follicle formation by regulating E-cadherin junctions between oocytes in mouse ovaries.


Academic skills of chronically unemployed drug-addicted adults.
The strong association between unemployment and drug addiction suggests that employment interventions are an important and needed focus of drug-addiction treatment. The increasing necessity of possessing basic academic skills to function in the workplace may require that some individuals receive educational training along with vocational training. This study investigated the academic skills of drug-addicted and chronically-unemployed adults (N = 559) who were enrolled in one of six studies conducted at the Center for Learning and Health in Baltimore, MD. Upon study enrollment, academic skills in math, spelling, and reading were examined using the Wide Range Achievement Test (WRAT-3 or WRAT-4) and educational history was examined using the Addiction Severity Index-Lite. Although participants completed an average of 11 years of education, actual academic skill level was at or below the seventh grade level for 81% of participants in math, 61% in spelling, and 43% in reading, and most participants were classified as Low Average or below based on age group norms. Despite the fact that participants in this analysis were studied across several years and were from diverse populations, rates of high school completion and academic skill levels were remarkably similar. Programs designed to improve the long-term employment status of drug-addicted individuals may benefit from the inclusion of basic adult education; future research on the topic is needed. Although establishing basic skills does not directly address chronic unemployment, it may help individuals obtain the jobs they desire and function effectively in those jobs.


Localization of 16 exons to a 450-kb region involved in the autoimmune polyglandular disease type I (APECED) on human chromosome 21q22.3.
As a step toward identifying the pathogenic genes for autoimmune polyglandular disease type I (APECED) and other disorders mapped to the PFKL locus on chromosome 21q22.3, we have constructed a cosmid/BAC (bacterial artificial chromosome) contig of 450 kb covering markers D21S1460-D21S25-PFKL-D21S154 and performed exon trapping. We isolated 22 distinct exons including 6 exons derived from two known genes (PFKL and EHOC-1). Among 16 novel exons, 2 exons matched with human expressed sequence tags (EST) and 7 exons showed homology at predicted amino acid sequence level with proteins from other species. These 16 exons were mapped back to the cosmid contigs, 12 of which were confirmed for their expression by polymerase chain reaction (PCR) screening of human cDNA libraries of various tissues. These exon sequences and a transcript map will aid for isolation of corresponding genes which will be identified as candidate genes involved in the pathogenesis of disorders mapped to the 21q22.3 region.


TUBULOINTERSTITIAL NEPHRITIS WITH UVEITIS (TINU SYNDROME). A CASE REPORT.
In this case report, we describe the case of a 50-year-old woman referred by her general practitioner to a pulmonologist in order to investigate persistent fever and elevation of C-reactive protein despite antibiotic treatment following a respiratory infection. The patient was examined extensively, during which rheumatology, gastroenterology, nephrology, ophthalmology, laboratory and imaging tests were performed. Due to a rapid progression of renal insufficiency with active urinary sediment, the patient was referred for a renal biopsy, which confirmed tubulointerstitial nephritis, followed by a diagnosis of bilateral anterior uveitis two months later - genetic testing was also conducted, which confirmed the diagnosis of tubulointerstitial nephritis with uveitis syndrome. Steroid treatment brought about a gradual reduction of proteinuria and a stabilisation of renal function.


Costing essential services package provided by a non-governmental organization network in Bangladesh.
The health profile of Bangladesh has improved remarkably, yet gaps in delivering quality health care remain. In response to the need for evidence to quantify resources for providing health services in Bangladesh, this study estimates unit costs of providing the essential services package (ESP) in the not-for-profit sector. This study used a stratified sampling approach to select 18 static clinics, which had fixed facilities, from 330 non-profit clinics under Smiling Sun network in Bangladesh. Costs were estimated from the providers' perspective, using both top-down and bottom-up methods, from July 2014 to June 2015. In total, there were 1115 observations (clients) for the 13 primary care services analysed. The estimated 2015 average costs per visit were: antenatal care ($7.03), postnatal care ($4.57), control of diarrheal diseases ($1.32), acute respiratory infection ($1.53), integrated management of child illness ($2.02), sexually transmitted infections ($4.70), reproductive tract infections ($3.56), tuberculosis ($41.65), limited curative care ($4.30), immunization ($2.23), family planning ($0.72), births by normal delivery ($29.45) and C-section ($114.83). Unit costs varied widely for each service, both between individual patients and among clinic level means. The coefficient of variation for the 13 services averaged 66%, implying potential inefficiencies. In addition, 32.9% of clients were not offered any lab test during the first antenatal visit. The unit cost of essential services differed by the type and location of clinics. Ultra clinics, on average, incurred 37% higher costs than vital (outpatient type) clinics, and urban clinics spent 40% more than rural clinics to deliver a unit of service. The study suggests that inefficiency and quality concerns exist in health service delivery in some facilities. Increasing the volume of clients through demand-side mechanisms and standardization of services would help address those concerns. Unit costs of services provide essential information for estimating resource needs for scaling up the ESPs.


Implementation of holistic nursing interventions based on fast track surgery concept in patients with lower extremity arterial occlusive disease.
Lower Extremity Arterial Occlusive Disease (LEAOD) is a prevalent condition affecting many patients worldwide, which requires careful management and patient cooperation. This study aimed to evaluate the effectiveness of holistic nursing interventions based on the Fast Track Surgery (FTS) concept in patients with LEAOD. A retrospective analysis of 92 LEAOD patients, randomized into control and experimental groups, was performed. Conventional rehabilitation nursing interventions were applied to the control group, while the experimental group received holistic rehabilitation nursing interventions based on the FTS concept. Patient adherence was assessed before and after the intervention using a hospital survey, and sleep quality was evaluated using the Pittsburgh Sleep Quality Index on days 3, 7, and 15 post-interventions. Post-intervention, the experimental group exhibited significantly improved adherence to balanced diet, regular exercise, timely medication, and regular review visits compared to the control group (P < .05). Further, Pittsburgh Sleep Quality Index scores indicated significantly better sleep quality over time in the experimental group than in the control group (P < .05). The implementation of holistic nursing interventions based on the FTS concept significantly improved patient adherence and sleep quality in LEAOD patients. These findings highlight the potential benefits of integrating such interventions in the management of LEAOD patients, potentially enhancing postoperative recovery and overall health outcomes.


Live long and prosper? Childhood living conditions, marital status, social class in adulthood and mortality during mid-life: a cohort study.
The aim of the present study was to investigate the impact of childhood living conditions, marital status, and social class in adulthood on the risk of mortality during mid-life. Two questions were addressed: Is there an effect of childhood living conditions on mortality risk during mid-life and if so, is the effect mediated or modified by social class and/or marital status in adulthood? A nationally representative, Swedish, level of living survey from 1968 was used as baseline. The study included those aged 25-69 at baseline (n = 4082). Social conditions in childhood and adulthood were assessed using self-reports. These individuals were then followed for 39 years using registry data on mortality. The results showed associations between childhood living conditions, marital status, social class in adulthood and mortality during mid life. Social class and familial conditions during childhood as well as marital status and social class in adulthood all contributed to the risk of mortality during mid-life. Individuals whose father's were manual workers, who grew up in broken homes, who were unmarried, and/or were manual workers in adulthood had an increased risk of mortality during mid life. The effects of childhood conditions were, in part, both mediated and modified by social class in adulthood. The findings of this study suggest that there are structural, social conditions experienced at different stages of the life course that affect the risk of mortality during mid-life.


[Experience with the clinical use of lithium oxybutyrate in patients with cardiac rhythm disorders].
An antiarrhythmic effect of lithium oxybutyrate was demonstrated in 81% of patients with ventricular extrasystole in the presence of affective disturbances, those with hypersympathicotonus in particular. In arrhythmic cases, lithium oxybutyrate was shown to have a hypokinetic effect on central hemodynamics and contribute to peripheral vasodilatation without causing clinically apparent circulatory disorders. The drug inhibits platelet aggregation and moderately increases blood procoagulation activity in the presence of greater erythrocyte deformability and improved bulbar conjunctival microcirculation.


Dietary compositions and their seasonal shifts in Japanese resident birds, estimated from the analysis of volunteer monitoring data.
Determining the composition of a bird's diet and its seasonal shifts are fundamental for understanding the ecology and ecological functions of a species. Various methods have been used to estimate the dietary compositions of birds, which have their own advantages and disadvantages. In this study, we examined the possibility of using long-term volunteer monitoring data as the source of dietary information for 15 resident bird species in Kanagawa Prefecture, Japan. The data were collected from field observations reported by volunteers of regional naturalist groups. Based on these monitoring data, we calculated the monthly dietary composition of each bird species directly, and we also estimated unidentified items within the reported foraging episodes using Bayesian models that contained additional information regarding foraging locations. Next, to examine the validity of the estimated dietary compositions, we compared them with the dietary information for focal birds based on stomach analysis methods, collected from past literatures. The dietary trends estimated from the monitoring data were largely consistent with the general food habits determined from the previous studies of focal birds. Thus, the estimates based on the volunteer monitoring data successfully detected noticeable seasonal shifts in many of the birds from plant materials to animal diets during spring-summer. Comparisons with stomach analysis data supported the qualitative validity of the monitoring-based dietary information and the effectiveness of the Bayesian models for improving the estimates. This comparison suggests that one advantage of using monitoring data is its ability to detect dietary items such as fleshy fruits, flower nectar, and vertebrates. These results emphasize the potential importance of observation data collecting and mining by citizens, especially free descriptive observation data, for use in bird ecology studies.


Radiation dose estimation by tooth enamel EPR dosimetry for residents of Dolon and Bodene.
The method of electron paramagnetic resonance (EPR) dosimetry was applied to the enamel of the teeth extracted from the residents of the Dolon and Bodene settlements of the Beskaragay district, which is the area adjacent to the radioactive fallout of the most contaminating nuclear test of 1949. The individual accidental radiation doses due to the fallout were obtained from the amplitude of the radiation induced EPR signal from the CO2- radical using the calibration method, after determining the parameters of EPR measurements to obtain the best reproducibility of the signal intensities. It was shown that after subtracting the natural background dose from the total absorbed dose obtained by EPR the residents of Dolon and Bodene received accidental radiation doses up to 356 mGy with an average value of 74.1+/-45.5 mGy before 1949 while the younger population received up to about 100 mGy with an average value of 11.5+/-37.7 mGy.


Vaccines for preventing cholera: killed whole cell or other subunit vaccines (injected).
Injected cholera vaccines are rarely used today, although they may have some benefit. It is valuable to summarize the evidence for effectiveness of injected cholera vaccines for comparison with newer oral vaccines (subject of a separate Cochrane Review). To evaluate killed whole cell (KWC) cholera vaccines and other inactive subunit vaccines (administered by injection) for preventing cholera and death, and to evaluate the adverse effects. In September 2008, we searched the Cochrane Infectious Diseases Group Specialized Register, CENTRAL (The Cochrane Library 2008, Issue 3), EMBASE, and LILACS. We also searched reference lists and handsearched the journal Vaccine up to 1997. Randomized and quasi-randomized controlled trials comparing injected cholera vaccines (KWC or other inactive subunit) with placebo, control vaccines, or no intervention in adults and children irrespective of immune status or special risk category. Two authors extracted data and assessed trial methodological quality independently. Dichotomous data were reported using the risk ratio (RR) with 95% confidence intervals (CI). Vaccine efficacies were also calculated (% vaccine efficacy = (1-RR) x 100%). Sixteen trials, involving over one million adults, children and infants, fulfilled the inclusion criteria. Twenty-four comparisons reported on vaccine efficacy (cholera cases and/or deaths) and 11 comparisons considered adverse effects (nine reported on both). Compared to placebo, vaccinees had a reduced risk of death from cholera (RR 0.49, 95% CI 0.25 to 0.93; 837,442 participants) and a reduced risk of contracting cholera at 12 months (RR 0.52, 95% CI 0.42 to 0.65, random-effects model; 1,512,573 participants). This translates to an efficacy of 48%, 95% confidence interval 35% to 58%. Significant protection lasted for two years, even after only a single dose, and for three years with an annual booster. Children over five years and adults were protected for up to three years, while children under five years were protected for up to a year. Injected cholera vaccines were associated with more systemic and local adverse effects compared to placebo, but these were not severe or life-threatening. Injected cholera vaccines appear to be safe and relatively more effective than usually realized. Protection against cholera persists for up to two years following a single dose of vaccine, and for three years with an annual booster. However, they have been superseded by oral vaccines.


Zinc transporter genes are coordinately expressed in men and women independently of dietary or plasma zinc.
The zinc transporter (ZnT; SLC30) and Zrt- and Irt-like protein (Zip, SLC39) zinc transporter families are integral to the maintenance of intracellular zinc concentrations. Few studies have examined the expression patterns of zinc transporter genes in human primary tissues. This study investigated the expression levels of a range of zinc transporter mRNA in the peripheral blood mononuclear cells of healthy men and women (n = 40) using quantitative real-time PCR. It also explored the relationships among zinc transporter expression levels, plasma zinc concentrations, and dietary zinc intake. The relative expression of the zinc transporter mRNA varied considerably, with ZnT7, ZnT1, and Zip1 being the most abundantly expressed. ZnT1 and Zip1 mRNA were highly correlated with one another (r = 0.9; P < 0.001) and with ZnT5, ZnT7, Zip3, and Zip10 (P < 0.001). When analyzed by gender, a correlation between the mRNA of ZnT7 and Zip3 (r = 0.6; P < 0.01) was demonstrated only in women. Zip10 mRNA was correlated with ZnT1 and Zip1 (r = 0.9; P < 0.001) in men only. In a regression analysis, plasma zinc variability was not significantly explained by dietary zinc intake, gender, age, or any individual or combination of zinc transporters. This study expands what is known about both the levels of zinc transporter gene transcription in humans and the extent of its variation in healthy men and women. The positive association between the mRNA of ZnT1 and Zip1, which have reciprocal roles in zinc transport across the plasma membrane, provides insight into the coordinated control of zinc homeostasis in humans.


The morphology and syntax of individuals who use AAC: research review and implications for effective practice.
A research review of 31 studies pertaining to the morphology and syntax of individuals who use augmentative and alternative communication (AAC) and who had severe speech and physical impairments (SSPI) was completed. Results suggest that, although many individuals who use AAC exhibit comprehension and expression of a wide range of grammatical structures, these individuals are at risk for experiencing grammar deficits. A wide range of individual differences was noted, both within and across many of the investigations. One of the more robust findings was that individuals tended to produce shorter utterances when they used graphic symbol-based AAC systems than would be expected, based on participant profiles; however, there appeared to be a developmental trend toward using longer utterances. There has been a dearth of research pertaining to the development and evaluation of appropriate assessment tools and intervention techniques to enhance grammar acquisition for individuals who use AAC. Considerations for developing such tools and techniques are discussed.


Photo-irradiated caffeic acid exhibits antimicrobial activity against Streptococcus mutans biofilms via hydroxyl radical formation.
An antimicrobial technique based on photo-oxidation of caffeic acid (CA) has recently been developed, but its effect on biofilm-forming bacteria is unknown. The present study aimed to evaluate the effect of photo-irradiated CA against Streptococcus mutans (cariogenic bacteria) biofilm as it relates to hydroxyl radical formation. S. mutans biofilms grown on hydroxyapatite disks were immersed in CA solution (0-2 mg/mL) and irradiated with LED light at wavelengths of 365, 385, and 400 nm and at irradiances of 500, 1000, and 2000 mW/cm2 for 4 min. Biofilm viable bacterial counts were determined by colony counting. The yield of hydroxyl radicals generated by the LED irradiation of CA solution was quantified by electron spin resonance analysis. Of the conditions tested, the highest bactericidal effect, with a > 5-log reduction in viable bacterial counts, was obtained by irradiation of a 1 mg/mL CA solution with 385 nm LED and at an irradiance of 2000 mW/cm2. Hydroxyl radical formation was related to this bactericidal effect. The present study suggests that the antimicrobial technique based on the 385 nm LED irradiation of CA is effective against cariogenic biofilms and can be applied as an adjunctive chemotherapy for dental caries.


Is severity of motor coordination difficulties related to co-morbidity in children at risk for developmental coordination disorder?
Aim of the study was to investigate whether 7-9 year old children with severe motor difficulties are more at risk of additional difficulties in activities in daily living, academic skills, attention and social skills than children with moderate motor difficulties. Children (N=6959) from a population based cohort, the Avon Longitudinal Study of Parents and Children (ALSPAC), were divided into three groups based on their scores on the ALSPAC Coordination Test at age 7: control children (scores above 15th centile; N=5719 [82.1%]); children with moderate (between 5th and 15th centile; N=951 [13.7%]); and children with severe motor difficulties (below 5th centile N=289 [4.2%]). Children with neurological disorders or an IQ<70 were excluded. Logistic regression was used to compare children with moderate and severe motor coordination difficulties with each other and with control children regarding their risk of co-morbidity defined as significant (<10th centile) difficulties with activities of daily living (ADL); academic skills (reading, spelling and handwriting); attention; social skills (social cognition and nonverbal skills). Children with severe motor difficulties demonstrated a higher risk of difficulties in ADL, handwriting, attention, reading, and social cognition than children with moderate motor difficulties, who in turn had a higher risk of difficulties than control children in five out of seven domains. Screening and intervention of co-morbid problems is recommended for children with both moderate and severe motor difficulties.


Mouthwash use and associated head and neck cancer risk.
All studies with questionnaire items on mouthwash use in the International Head and Neck Cancer Epidemiology Consortium (INHANCE). Pooled analysis data from case controlled studies using Individual Patient Data (IPD) meta-analysis methods. Logistic regression was used to assess the association of mouthwash use with cancers of the oral cavity, oropharynx, hypopharynx and larynx, adjusting for study, age, sex, pack-years of tobacco smoking, number of alcoholic drinks/day and education. Eight thousand, nine hundred and eighty-one cases of head and neck cancer and 10,090 controls from 12 case-control studies with comparable information on mouthwash use were included in the analysis. Compared with never users of mouthwash, the odds ratio (OR) of all head and neck cancers was 1.01 [95% confidence interval (CI): 0.94-1.08] for ever users, based on 12 studies. The corresponding ORs of cancer of the oral cavity and oropharynx were 1.11 (95% CI: 1.00-1.23) and 1.28 (95% CI: 1.06-1.56), respectively. Although limited by the retrospective nature of the study and the limited ability to assess risks of mouthwash use in nonusers of tobacco and alcohol, this large investigation shows potential risks for head and neck cancer subsites and in long-term and frequent users of mouthwash.


MexY-promoted aminoglycoside resistance in Pseudomonas aeruginosa: involvement of a putative proximal binding pocket in aminoglycoside recognition.
The resistance-nodulation-division (RND) family multidrug efflux system MexXY-OprM is a major determinant of aminoglycoside resistance in Pseudomonas aeruginosa, although the details of aminoglycoside recognition and export by MexY, the substrate-binding RND component of this efflux system, have not been elucidated. To identify regions/residues of MexY important for aminoglycoside resistance, plasmid-borne mexY was mutagenized and mutations that impaired MexY-promoted aminoglycoside (streptomycin) resistance were identified in a ΔmexY strain of P. aeruginosa. Sixty-one streptomycin-sensitive mexY mutants were recovered; among these, 7 unique mutations that yielded wild-type levels of MexY expression were identified. These mutations compromised resistance to additional aminoglycosides and to other antimicrobials and occurred in both the transmembrane and periplasmic regions of the protein. Mapping of the mutated residues onto a 3-dimensional structure of MexY modeled on Escherichia coli AcrB revealed that these tended to occur in regions implicated in general pump operation (transmembrane domain) and MexY trimer assembly (docking domain) and, thus, did not provide insights into aminoglycoside recognition. A region corresponding to a proximal binding pocket connected to a periplasm-linked cleft, part of a drug export pathway of AcrB, was identified in MexY and proposed to play a role in aminoglycoside recognition. To test this, selected residues (K79, D133, and Y613) within this pocket were mutagenized and the impact on aminoglycoside resistance was assessed. Mutations of D133 and Y613 compromised aminoglycoside resistance, while, surprisingly, the K79 mutation enhanced aminoglycoside resistance, confirming a role for this putative proximal binding pocket in aminoglycoside recognition and export. IMPORTANCE Bacterial RND pumps do not typically accommodate highly hydrophilic agents such as aminoglycosides, and it is unclear how those, such as MexY, which accommodate these unique substrates, do so. The results presented here indicate that aminoglycosides are likely not captured and exported by this RND pump component in a unique manner but rather utilize a previously defined export pathway that involves a proximal drug-binding pocket that is also implicated in the export of nonaminoglycosides. The observation, too, that a mutation in this pocket enhances MexY-mediated aminoglycoside resistance (K79A), an indication that it is not optimally designed to accommodate these agents, lends further support to earlier proposals that antimicrobials are not the intended pump substrates.


[p-Sulfotetrafluorophenyl hydrophilic activated esters of amino acids in peptide synthesis].
Highly reactive hydrophilic (i.e., water-soluble) p-sulfotetrafluorophenyl esters (Tfs esters) are proposed for peptide synthesis in aqueous and aqueous-organic media, as well as for protein and peptide partial synthesis in an aqueous medium. These esters can serve as a basis for creating a series of protein modifying reagents. As they are analogs of the widely used pentafluorophenyl esters, the Tfs esters possess a high reactivity coupled with good stability during storage. The expression for the reaction rate (for substrates AA1 and AA2) is shown to be v = k[Boc-AA1-OTfs][H-AA2-NH2]0.5 for both water and DMF, i.e., the reaction is not a simple second-order reaction. The reaction rate in water is only slightly lower than that in DMF.


Primary liver cell carcinoma associated with infective liver disease.
Three patients, who had primary liver cell carcinoma combined with infective disorders (one each with pyogenic liver abscess, hepatic tuberculosis and amoebic liver abscess), are presented. The importance of further investigation, particularly liver biopsy and/or peritoneoscopy, in patients with liver diseases is highlighted.


Fucoxanthin-Rich Brown Algae Extract Improves Male Reproductive Function on Streptozotocin-Nicotinamide-Induced Diabetic Rat Model.
Hypogonadism and oxidative stress are occurring commonly in men with diabetes and associated male infertility. This study aimed to investigate the capability of anti-oxidative and anti-inflammatory properties of fucoxanthin as well as to evaluate its protective effects on male reproduction in diabetic rats. The RAW 264.7 macrophage cells were used to evaluate the anti-oxidative and anti-inflammatory activity. Thirty male Sprague-Dawley rats were induced by streptozotocin-nicotinamide for a diabetes model and fed either with three different doses of fucoxanthin (13, 26, and 65 mg/kg) or rosiglitazone (0.571 mg/kg) for four weeks. The fucoxanthin significantly inhibited nitric oxide production and reduced reactive oxygen species level in lipopolysaccharide-induced RAW 264.7 cells. In the animal study, fucoxanthin administration improved insulin resistance, restored sperm motility, decreased abnormal sperm number, and inhibited lipid peroxidation. Moreover, it restored GPR54 and SOCS-3 mRNA expression in the hypothalamus and recovered luteinizing hormone level, as well as the testosterone level. In conclusion, fucoxanthin not only possessed antioxidant and anti-inflammatory properties but also decreased the diabetes signs and symptoms as well as improved spermatogenesis and male reproductive function.


The implications of urine drug testing in pain management.
In the treatment of pain management, physicians employ a variety of drugs, ranging from low-impact to highly potent, and to maximize patient health, urine toxicology analyses can significantly improve the delivery of pain treatment. Drugs such as opioids that are used for pain management are peculiar in that they provide effective pain relief and have a high risk of addiction. The use of illicit drugs in the general population has been on the rise; however, self-reporting and close monitoring of patient behavior are insufficient means to detect drug abuse and confirm compliance. Therefore, in order to create more effective drug treatment plans, physicians must understand and account for the implications of patient drug use history. Urine toxicology analysis is an important tool for pain physicians because it is more sensitive than most alternative blood tests, more efficient and cost-effective. Urine testing in addition to improving patient pain management also has forensic and legal implications. There are however limitations to urine toxicology methods as they can produce false-positive and false-negative results and are prone to human error and sample contamination There is also a need for more specific and rapid urine drug testing. Healthcare professionals should therefore be familiar with the limitations of various urine drug testing methods, and possess skills necessary to properly interpret these results. This review suggests that the overall benefits incurred by both the patient's short-term and long-term health support the routine integration of urine toxicology analysis in routine clinical care. In addition to improving health care and patient health, it has a strong potential to improve patient-physician relationships and protects the interest of involved healthcare practitioners.


Identification and preliminary characterization of external membrane-bound nuclease activities in Mycoplasma pulmonis.
Mycoplasma pulmonis has substantial DNase activity exposed on the cell surface. At least part of this activity is attributable to an endonuclease. The activity is destroyed at 56 degrees C and inhibited by either 5 mM EDTA or 10 mM zinc chloride. It can also be eliminated by treatment of intact organisms with trypsin and is regenerated by incubation of the treated organisms in a medium that supports protein synthesis. DNase exposed at the cell surface constitutes 20% of the total DNase activity present in M. pulmonis extracts.


Diversity in Infection Specificity between the Bloom-forming Microalga Heterosigma akashiwo and Its dsDNA Virus, Heterosigma akashiwo Virus.
Heterosigma akashiwo virus (HaV) is a dsDNA virus that infects the bloom-forming raphidoflagellate Heterosigma akashiwo. Both the host and its virus are phenotypically diverse in terms of infection specificity. Their relationships have been examined based on the occurrence or absence of algal lysis following virus inoculation; however, variations in the strain-level host-virus relationship regarding infectivity and lysis rates remain unclear. Therefore, we performed a series of cross-infectivity tests using 60 H. akashiwo and 22 HaV strains isolated from the coastal waters of western Japan. The host strains were divided into 5 different groups and viruses into 4 groups. Using a representative strain from each group, algal lysis was observed in 14 of the (5×4=) 20 host-virus combinations; the concentration of infectious units in each HaV suspension was then assessed using the most probable number (MPN) assay on the five host strains. Virus titers ranged between 1.1×101 and 2.1×107 infectious units mL-1; the titer of each viral lysate was differently estimated using distinct H. akashiwo strains as hosts. These results suggest that (1) a clonal viral lysate comprises virions with different intraspecific infection specificities and/or (2) the efficiency and error rates of each intracellular replication process vary in each host-virus combination.


Controlled trial of continuous inflating pressure for hyaline membrane disease.
A controlled trial of elective intervention with continuous inflating pressure (CIP) was performed in infants with severe hyaline membrane disease who weighed more than 1000 g at birth. Infants entered the trial if their arterial oxygen tension (PaO2) fell below 60 mmHg while breathing a fractional inspired oxygen concentration (F1O2) greater than 0-95. 11 out of 12 infants in the CIP-treated group and 10 out of 12 in the control group survived. 7 treated and 6 control infants required mechanical ventilation. When CIP was started the Pao2 of the treated infants increased, and they breathed high concentrations of oxygen for a significantly shorter period than the control infants. During the 31-month duration of the trial 107 other infants with severe hyaline membrane disease were admitted who did not meet the criteria for entry to the trial. 37 survived after breathing high concentrations of oxygen (F1O2 greater than 0-60) spontaneously without any ventilatory assistance, and the remaining 70 infants were already being ventilated on their arrival in the unit, usually because they had required mechanical ventilation during transfer from other hospitals. The neonatal survival rate for those infants born in this hospital during the study period was 88% (50 out of 57 infants) and for those referred from other hospitals it was 69% (51 out of 74 infants). The maximum further increase in overall survival rate that might have been achieved in our population of infants if CIP had been initiated very early in the course of the illness was 5%--i.e. from 77% (101/131) to 82% (107/131).


IGF1 stimulates greater muscle hypertrophy in the absence of myostatin in male mice.
Insulin-like growth factors (IGFs) and myostatin have opposing roles in regulating the growth and size of skeletal muscle, with IGF1 stimulating, and myostatin inhibiting, growth. However, it remains unclear whether these proteins have mutually dependent, or independent, roles. To clarify this issue, we crossed myostatin null (Mstn-/-) mice with mice overexpressing Igf1 in skeletal muscle (Igf1+) to generate six genotypes of male mice; wild type (Mstn+/+ ), Mstn+/-, Mstn-/-, Mstn+/+:Igf1+, Mstn+/-:Igf1+ and Mstn-/-:Igf1+ Overexpression of Igf1 increased the mass of mixed fibre type muscles (e.g. Quadriceps femoris) by 19% over Mstn+/+ , 33% over Mstn+/- and 49% over Mstn-/- (P < 0.001). By contrast, the mass of the gonadal fat pad was correspondingly reduced with the removal of Mstn and addition of Igf1 Myostatin regulated the number, while IGF1 regulated the size of myofibres, and the deletion of Mstn and Igf1+ independently increased the proportion of fast type IIB myosin heavy chain isoforms in T. anterior (up to 10% each, P < 0.001). The abundance of AKT and rpS6 was increased in muscles of Mstn-/-mice, while phosphorylation of AKTS473 was increased in Igf1+mice (Mstn+/+:Igf1+, Mstn+/-:Igf1+ and Mstn-/-:Igf1+). Our results demonstrate that a greater than additive effect is observed on the growth of skeletal muscle and in the reduction of body fat when myostatin is absent and IGF1 is in excess. Finally, we show that myostatin and IGF1 regulate skeletal muscle size, myofibre type and gonadal fat through distinct mechanisms that involve increasing the total abundance and phosphorylation status of AKT and rpS6.


The physiological function of lower urinary tract smooth muscle.
The lower urinary tract is a muscular system composed of the urinary bladder and the outflow tract. During filling with urine the bladder is relaxed and the outflow tract offers a high resistance; during emptying the outflow resistance falls and the bladder wall generates a high wall tension to raise intravesical pressure. The coordination of these responses is organized in the brainstem and sacral spinal cord to control the activity of autonomic and somatic efferents to the smooth muscle of the bladder (detrusor) and the smooth and skeletal muscle of the bladder base and urethra. Detrusor contraction is predominantly controlled by parasympathetic fibres releasing acetylcholine and ATP; the outflow tract is controlled by parasympathetic and sympathetic fibres to the bladder base (trigone) and urethral smooth muscle (including a nitregic component) and somatic fibres to the urethral rhabdosphincter. The smooth muscles also develop spontaneous contractions that determine the tone of the musculature. The cellular signaling pathways that evoke contraction due to neurotransmitter release and the origin of spontaneous activity are discussed, as well as the electrical properties of the smooth muscle relevant to the propagation of electrical signals. Finally the interaction of muscle cells with other cell types (epithelium and interstitial cells) is considered, relevant to their ability to regulate muscle contractility. Throughout, the basic physiological processes are considered in relation to pathological developments that are prevalent in the human lower urinary tract, in particular the overactive bladder and urinary incontinence, and the identification of drug targets to manage these conditions.


Stochasticity in physiologically based kinetics models: implications for cancer risk assessment.
In case of low-dose exposure to a substance, its concentration in cells is likely to be stochastic. Assessing the consequences of this stochasticity in toxicological risk assessment requires the coupling of macroscopic dynamics models describing whole-body kinetics with microscopic tools designed to simulate stochasticity. In this article, we propose an approach to approximate stochastic cell concentration of butadiene in the cells of diverse organs. We adapted the dynamics equations of a physiologically based pharmacokinetic (PBPK) model and used a stochastic simulator for the system of equations that we derived. We then coupled kinetics simulations with a deterministic hockey stick model of carcinogenicity. Stochasticity induced substantial modifications relative to dose-response curve, compared with the deterministic situation. In particular, there was nonlinearity in the response and the stochastic apparent threshold was lower than the deterministic one. The approach that we developed could easily be extended to other biological studies to assess the influence of stochasticity at macroscopic scale for compound dynamics at the cell level.


[Choice of treatment in patients with urolithiasis of anomalous kidneys].
We studied efficacy of transcutaneous nephrolithotripsy (TNLT) in patients with urolithiasis and anomalous upper urinary tract. For two years we performed TNLT in 148 patients with complicated forms of urolithiasis and anomalous kidneys (age 18-86, 76 females and 72 males). The examination for urolithiasis has found different anomalies of the urinary tract in these patients: double pelvis and ureter (9 cases--6.1%), lumbar dystopy of the kideney (59 cases--39.9%), horseshoe kidney (12 cases--8.1%), solitary cyst of the kidney (43 cases--29.0%), parapelvic cysts (25 cases--16.9%). Extracorporeal shock-wave lithotripsy (ESWL) was conducted as first-line treatment in the other 139 patients with nephroliths under 1.5-2 cm in size who had the following anomalies of the urinary system: a solitary kidney (n=14), double pelvis and ureter (n=26), anomalous position of the kidney (n=46), anomalous shape of the kidney (n=20), simple and parapelvic cysts (n=33). A total of 154 TNLT were performed in 148 patients. Transcutaneous interventions were most frequent in the treatment of coral (31.3%) and pelvoureteral (14.6%) concrements, one-stage operation was made in 143 patients. Transcutaneous puncture nephrostomy as the first stage was made in 5 patients. The concrements were removed by one operative intervention in 132 (89.2%) out of 148 patients. Exacerbation of pyelonephritis as a postoperative complication was treated conservatively in 12.5% patients. Residual fragments were diagnosed in 16 patients. Of them, 11 patients had multiple and 5 patients had coral concrements. Residual fragments were removed in 14 patients either by renephroscopy or by ESWL. Treatment of 139 patients with concrements less than 1.5-3 cm in size consisted in 197 ESWL sessions. In combination of anomaly and urolithiasis the number of lithotripsy sessions per one patient was 1.8 +/- 0.2. The least number of ESWL sessions per one patient was conducted in patients with lumbar dystopy of the kidney (1.3 +/- 0.45), the greatest number in horseshoe kidney (3.8 +/- 0.8). Evacuation of the concrements after ESWL depended on the type of anomaly and concrement location. It was 61-69% in patients with anomaly of the shape and position of the kidney, in cystic anomalies is was 89-91%. Open surgery was indicated in 7 patients who needed plastic surgery of hydronephrosis (4) and obstructive megaureter (3). Thus, low-invasive operations provide high efficacy of urolith removal in patients with anomalies of the urinary tract and can replace traumatic open operations.


Immunochemical characterization and antifertility effect of an androgen independent antigen of spermatozoa identified by a monoclonal antibody (D7G3).
Monoclonal antibody, (mAb) D7G3, directed to human spermatozoa and cross-reactive with mouse and rat spermatozoa was found to be a sperm agglutinating antibody. It reacted with antigen present on the post acrosomal region of human spermatozoa and acrosomal region of mouse spermatozoa. The antigen reacting with mAb D7G3 was localized in mouse testicular germ cells and Sertoli cells. It was also localized in the epithelium and spermatozoa of the caput, corpus and cauda epididymides. In addition, the antigen was detected in the epithelial cells and secretions of the prostate and seminal vesicle. The mAb D7G3 reacted with a band of molecular mass 16 kDa in testicular and epididymal sperm and protein preparations of ventral prostate. In the seminal vesicle, an additional band of molecular mass 36 kDa was also identified. The effect of castration on the expression of proteins was studied in the rat. Immunohistochemical localization and Western blotting experiments showed that the antigens identified by the mAb D7G3 was detectable in the epididymis, ventral prostate and seminal vesicle after two weeks of castration. In the seminal vesicle, three additional bands were identified by mAb D7G3 following castration. The expression of this antigen throughout spermatogenesis and sperm maturation indicated that it may have an important biological function. A polyclonal antiserum directed to the 16 kDa mouse epididymal sperm protein was raised in rabbits. Passive immunization of female mice with this antibody caused a significant reduction in fertility only if administered 24 hours prior to mating, indicating that the antibody inhibited a pre-fertilization event by inhibiting sperm function.


Characterization of piRNAs across postnatal development in mouse brain.
PIWI-interacting RNAs (piRNAs) are responsible for maintaining the genome stability by silencing retrotransposons in germline tissues- where piRNAs were first discovered and thought to be restricted. Recently, novel functions were reported for piRNAs in germline and somatic cells. Using deep sequencing of small RNAs and CAGE of postnatal development of mouse brain, we identified piRNAs only in adult mouse brain. These piRNAs have similar sequence length as those of MILI-bound piRNAs. In addition, we predicted novel candidate regulators and putative targets of adult brain piRNAs.


Prior depth information can bias motion perception.
Previous studies of the motion aperture problem have shown that the direction of grating motion can be biased by using binocular disparity to designate borders of the aperture as intrinsic (belonging to the grating) or extrinsic (resulting from occlusion of the grating). Observers report motion in the direction of the extrinsic border, as if the grating was extended and moving underneath an occluding surface. Here we investigate whether prior information about depth ordering, given by structure-from-motion, can bias the perceived motion direction of a subsequent moving grating in a similar manner. We presented an aperture stimulus that rotated about its vertical and horizontal axes, revealing the depth relationships (intrinsic and extrinsic) of the aperture borders. The grating then translated within the aperture and observers reported the direction of perceived motion. The test stimulus contained no information about the depth ordering of the scene. We found that observers' reported motion shifted toward the direction of the occluding edges, consistent with the intrinsic-extrinsic border predictions. These results indicate that prior scene information, not just depth information explicitly defined in the test stimulus, is used to help solve the motion aperture problem.


PEGylation is effective in reducing immunogenicity, immunotoxicity, and hepatotoxicity of α-momorcharin in vivo.
α-momorcharin (α-MMC), a type I ribosome-inactivating protein (RIP) from Momordica charantia, is well known for its antitumor and antivirus activities. However, the immunotoxicity and hepatotoxicity hampers its potential therapeutic usage. In order to reduce its toxicity, we had modified the α-MMC with polyethylene glycol (PEG), and detected the toxicity of the PEGylated α-MMC conjugates (α-MMC-PEG) in vivo. After α-MMC purified from bitter melon seeds, α-MMC-PEG was constructed with a branched 20 kDa (mPEG) 2-Lys-NHS, the tests of immunogenicity, immunotoxicity, and general toxicity of α-MMC-PEG were conducted in guinea pig and rat. The titer of specific IgG in rats, immunized by α-MMC-PEG, were approximately one-third of those that by α-MMC, all the guinea pigs treated with α-MMC died of anaphylaxis shock within 5 min, while no animals treated with α-MMC-PEG died in the active systemic anaphylaxis (ASA) test. The passive cutaneous anaphylaxis (PCA) reaction of α-MMC-PEG challenge in rats was significantly smaller than that of the α-MMC. The liver damage was greatly released, such as the change of globulin (GLB), aspartate aminotransferase (AST), total bilirubin (TBIL) cholesterol (CHOL), albumin (ALB), and the degree of hepatocyte necrosis in repeated toxicity study. PEGylation is effective in reducing the immunogenicity, immunotoxicity, and hepatotoxicity of α-MMC in vivo.


Quality care in transverse myelitis: a responsive protocol.
This study was conducted to aid in the development of a multidisciplinary care center for patients with transverse myelitis. We surveyed the parents of 20 children diagnosed with transverse myelitis between the ages of 0.5 and 21 years to understand their experiences in navigating the health care system. We analyzed acute care events and long-term follow-up in relation to patient satisfaction. Results showed satisfactory ratings in the vicinity of 50% in key areas such as the articulation of a treatment plan. A significant disparity was found in the patients' desire for specialty care and their ability to procure such care. In all, 90% of patients expressed a desire to consult with a psychiatrist, but only 25% were successful in making a visit, a 64% deficit; 70% of respondents also desired to see a gastroenterologist, with only 25% actually doing so, leaving a 43% gap. Recommendations and patient opinions regarding the creation of a collaborative care environment are noted. Research with a larger sample will further elucidate the needs in transverse myelitis patient care.


Early systemic microvascular damage in pigs with atherogenic diabetes mellitus coincides with renal angiopoietin dysbalance.
Diabetes mellitus (DM) is associated with a range of microvascular complications including diabetic nephropathy (DN). Microvascular abnormalities in the kidneys are common histopathologic findings in DN, which represent one manifestation of ongoing systemic microvascular damage. Recently, sidestream dark-field (SDF) imaging has emerged as a noninvasive tool that enables one to visualize the microcirculation. In this study, we investigated whether changes in the systemic microvasculature induced by DM and an atherogenic diet correlated spatiotemporally with renal damage. Atherosclerotic lesion development was triggered in streptozotocin-induced DM pigs (140 mg/kg body weight) by administering an atherogenic diet for approximately 11 months. Fifteen months following induction of DM, microvascular morphology was visualized in control pigs (n = 7), non-diabetic pigs fed an atherogenic diet (ATH, n = 5), and DM pigs fed an atherogenic diet (DM+ATH, n = 5) using SDF imaging of oral mucosal tissue. Subsequently, kidneys were harvested from anethesized pigs and the expression levels of well-established markers for microvascular integrity, such as Angiopoietin-1 (Angpt1) and Angiopoietin-2 (Angpt2) were determined immunohistochemically, while endothelial cell (EC) abundance was determined by immunostaining for von Willebrand factor (vWF). Our study revealed an increase in the capillary tortuosity index in DM+ATH pigs (2.31±0.17) as compared to the control groups (Controls 0.89±0.08 and ATH 1.55±0.11; p<0.05). Kidney biopsies showed marked glomerular lesions consisting of mesangial expansion and podocyte lesions. Furthermore, we observed a disturbed Angpt2/Angpt1 balance in the cortex of the kidney, as evidenced by increased expression of Angpt2 in DM+ATH pigs as compared to Control pigs (p<0.05). In the setting of DM, atherogenesis leads to the augmentation of mucosal capillary tortuosity, indicative of systemic microvascular damage. Concomitantly, a dysbalance in renal angiopoietins was correlated with the development of diabetic nephropathy. As such, our studies strongly suggest that defects in the systemic microvasculature mirror the accumulation of microvascular damage in the kidney.


[Dietary assessment of adults in three villages in Warsaw, Radom and Biała Podlaska districts].
The objectives of this study was the assessment of dietary patterns of adults individuals living in three villages: Celestynów (Warsaw district), Jasionka (Biała podlaska district) and Babin (Radom district) in Poland. The study was conducted in winter 1994/95 among 71 men and 77 women aged 18 years or more. The information on food consumption during one weekday was obtained by 24-hour dietary recall method. On the average the percentage of total energy intake from proteins and fats was above the recommended limits whereas the proportion of energy derived from carbohydrates was slightly deficient. For women the average daily intake of most nutrients under study (with exception of animal proteins, fats, sodium, phosphorus, manganese, vitamin E, B2 and niacin) was below recommended levels. The consumption of carbohydrates (also dietary fiber), magnesium, zinc, copper, calcium was 29-35% below recommendations and case of vitamin C-51%. In general men had higher mean intake of energy and nutrients than women. Their average daily diet did not meet recommendation only for carbohydrates, dietary fiber, calcium magnesium, copper, vitamin C and A. Men consumed too much animal proteins, sodium, phosphorus and cholesterol. Daily consumption of minerals and vitamins from diet and supplements together for those subjects who used supplements (41% of respondents) met or exceed recommended dietary intake, with exception of potassium for women. Taking consideration number of faults founded in the nutrition patterns of adults from rural area more nutritional study in this subpopulation group should be conducted.


Anxiety symptoms and risk of cognitive decline in older community-dwelling men.
Previous research regarding anxiety as a predictor of future cognitive decline in older adults is limited and inconsistent. We examined the independent relationship between anxiety symptoms and subsequent cognitive decline. We included 2,818 community-dwelling older men (mean age = 76.1, SD ±5.3 years) who were followed on an average for 3.4 years. We assessed anxiety symptoms at baseline using the Goldberg Anxiety Scale (GAS; range = 0-9). We assessed cognitive function at baseline and at two subsequent visits using the Modified Mini-Mental State Examination (3MS; global cognition) and the Trails B test (executive function). At baseline, there were 690 (24%) men with mild anxiety symptoms (GAS 1-4) and 226 (8%) men with moderate/severe symptoms (GAS 5-9). Men with anxiety symptoms were more likely to have depressed mood, poor sleep, more chronic medical conditions, and more impairment in activities of daily living compared to those with no anxiety symptoms. Compared to those with no anxiety symptoms at baseline, men with any anxiety symptoms were more likely to have substantial worsening in Trails B completion time (OR = 1.56, 95% CI 1.19, 2.05). The association was attenuated after adjusting for potential confounders, including depression and poor sleep, but remained significant (OR = 1.40, 95% CI 1.04, 1.88). In cognitively healthy older men, mild anxiety symptoms may potentially predict future decline in executive functioning. Anxiety is likely a manifestation of an underlying neurodegenerative process rather than a cause.


Diatom-driven recolonization of microbial mat-dominated siliciclastic tidal flat sediments.
Modern microbial mats and biofilms play a paramount role in sediment biostabilization. When sporadic storms affect tidal flats of Bahía Blanca Estuary, the underlying siliciclastic sediment is exposed by physical disruption of the mat, and in a few weeks' lapse, a microbial community re-establishes. With the objective of studying colonization patterns and the ecological succession of microorganisms at the scale of these erosional structures, these were experimentally made and their biological recolonization followed for 8 weeks, with replication in winter and spring. Motile pennate diatoms led the initial colonization following two distinct patterns: a dominance by Cylindrotheca closterium in winter and by naviculoid and nitzschioid diatoms in spring. During the first 7 days, cell numbers increased 2- to 17-fold. Cell densities further increased exhibiting sigmoidal community growth, reaching 2.9-8.9 × 106 cells cm-3 maxima around day 30; centric diatoms maintained low densities throughout. In 56 days after removal of the original mat, filamentous cyanobacteria that dominate mature mats did not establish a significant biomass, leading to the rejection of the hypothesis that cyanobacteria would drive the colonization. The observed dominance of pennate diatoms is attributed to extrinsic factors determined by tidal flooding, and intrinsic ones, e.g. motility, nutrient affinity and high growth rate.


Regional European differences in allele and genotype frequencies of low density lipoprotein receptor-related protein 1 polymorphism in Alzheimer's disease.
The low density lipoprotein receptor-related protein 1 (LRP1 gene) is a candidate gene for Alzheimer's disease (AD), because it is a ligand for proteins involved in AD pathogenesis, such as apolipoprotein E (APOE), alpha2-macroglobulin (A2M), amyloid precursor protein (APP), and is located on chromosome 12, within a region linked with AD. An association between a silent polymorphism (C/T) in exon 3 and late onset AD has been reported, with an increased frequency of the C allele, although with conflicting results. We examined this polymorphism in a cohort of 166 sporadic AD patients and 225 sex- and age-matched nondemented controls from Southern Italy. No statistically significant differences were found in LRP1 genotype and allele frequencies between the whole AD sample and controls, nor in early- and late-onset subsets of AD patients. No statistically significant differences in frequencies between LRP1 alleles and AD among APOE allele, age, or gender strata were found. Finally, comparing our results with the findings from other European populations, the LRP1 C allele frequency showed a statistically significant decreasing trend from Northern to Southern regions of Europe, with a concomitant increase in LRP1 T allele frequency, but in AD patients only. Finally, in the AD sample, a decreasing geographical trend from North to South of Europe was found for LRP1 CC genotype, and an inverse trend for LRP1 CT genotype frequency. We suggest that these regional variations in LRP1 genotype and allele frequencies in AD could be related to the different patterns of association between this polymorphism and the disease in various European studies.


Needle-free delivery of measles virus vaccine to the lower respiratory tract of non-human primates elicits optimal immunity and protection.
Needle-free measles virus vaccination by aerosol inhalation has many potential benefits. The current standard route of vaccination is subcutaneous injection, whereas measles virus is an airborne pathogen. However, the target cells that support replication of live-attenuated measles virus vaccines in the respiratory tract are largely unknown. The aims of this study were to assess the in vivo tropism of live-attenuated measles virus and determine whether respiratory measles virus vaccination should target the upper or lower respiratory tract. Four groups of twelve cynomolgus macaques were immunized with 104 TCID50 of recombinant measles virus vaccine strain Edmonston-Zagreb expressing enhanced green fluorescent protein. The vaccine virus was grown in MRC-5 cells and formulated with identical stabilizers and excipients as used in the commercial MVEZ vaccine produced by the Serum Institute of India. Animals were immunized by hypodermic injection, intra-tracheal inoculation, intra-nasal instillation, or aerosol inhalation. In each group six animals were euthanized at early time points post-vaccination, whereas the other six were followed for 14 months to assess immunogenicity and protection from challenge infection with wild-type measles virus. At early time-points, enhanced green fluorescent protein-positive measles virus-infected cells were detected locally in the muscle, nasal tissues, lungs, and draining lymph nodes. Systemic vaccine virus replication and viremia were virtually absent. Infected macrophages, dendritic cells and tissue-resident lymphocytes predominated. Exclusive delivery of vaccine virus to the lower respiratory tract resulted in highest immunogenicity and protection. This study sheds light on the tropism of a live-attenuated measles virus vaccine and identifies the alveolar spaces as the optimal site for respiratory delivery of measles virus vaccine.


The manifold nature of interpersonal relations: the quest for a common mechanism.
It has been proposed that the capacity to code the 'like me' analogy between self and others constitutes a basic prerequisite and a starting point for social cognition. It is by means of this self/other equivalence that meaningful social bonds can be established, that we can recognize others as similar to us, and that imitation can take place. In this article I discuss recent neurophysiological and brain imaging data on monkeys and humans, showing that the 'like me' analogy may rest upon a series of 'mirror-matching' mechanisms. A new conceptual tool able to capture the richness of the experiences we share with others is introduced: the shared manifold of intersubjectivity. I propose that all kinds of interpersonal relations (imitation, empathy and the attribution of intentions) depend, at a basic level, on the constitution of a shared manifold space. This shared space is functionally characterized by automatic, unconscious embodied simulation routines.