The metabolic syndrome of omega3-depleted rats. I. Liver data.
Second-generation rats depleted in long-chain polyunsaturated omega3 fatty acids were recently proposed as a novel animal model for the metabolic syndrome. In the present study, a dietary deprivation of omega3 acids for 3-7 months was found sufficient to provoke in 6-week-old normal rats the same alteration of the fatty acid content and profile of liver phospholipids and triglycerides as that otherwise prevailing in the second-generation omega3-depleted rats, with emphasis on a severe decrease in their omega3 fatty acid content, alterations in the relative contribution of and ratio between selected long-chain polyunsaturated omega6 fatty acids, saturated and monodesaturated fatty acids and precursors of nervonic acid, and liver steatosis. When the omega3-depleted rats were exposed, after the first 7 months of the present experiments and for 2-4 weeks to a diet supplemented with 5% (w/w) flaxseed oil, most of these hepatic variables returned towards or beyond control values. In both the omega3-depleted rats and control animals, however, the eventual exposure to the flaxseed oil-enriched diet failed to suppress liver steatosis and, on the contrary, provoked a further increase in liver triglyceride content. It is proposed, therefore, that the present approach represents a simple and realistic animal model to study the consequences of omega3-depletion. Moreover, the results suggest that to oppose such consequences, e.g. liver steatosis, it may be necessary to combine the dietary supply of omega3 acids with a suitable control of food intake, in both qualitative and quantitative terms.


Cleavage of p75 neurotrophin receptor by alpha-secretase and gamma-secretase requires specific receptor domains.
The p75 neurotrophin receptor (p75(NTR)), a member of the tumor necrosis factor superfamily of receptors, undergoes multiple proteolytic cleavage events. These events are initiated by an alpha-secretase-mediated release of the extracellular domain followed by a gamma-secretase-mediated intramembrane cleavage. However, the specific determinants of p75(NTR) cleavage events are unknown. Many other substrates of gamma-secretase cleavage have been identified, including Notch, amyloid precursor protein, and ErbB4, indicating there is broad substrate recognition by gamma-secretase. Using a series of deletion mutations and chimeric receptors of p75(NTR) and the related Fas receptor, we have identified domains that are essential for p75(NTR) proteolysis. The initial alpha-secretase cleavage was extracellular to the transmembrane domain. Unfortunately, deletion mutants were not capable of defining the requirements of ectodomain shedding. Although this cleavage is promiscuous with respect to amino acid sequence, its position with respect to the transmembrane domain is invariant. The generation of chimeric receptors exchanging different domains of noncleavable Fas receptor with p75(NTR), however, revealed that a discrete domain above the membrane is sufficient for efficient cleavage of p75(NTR). Mass spectrometric analysis confirmed the cleavage can occur with a truncated p75(NTR) displaying only 15 extracellular amino acids in the stalk region.


The ACTA PORT-score for predicting perioperative risk of blood transfusion for adult cardiac surgery.
A simple and accurate scoring system to predict risk of transfusion for patients undergoing cardiac surgery is lacking. We identified independent risk factors associated with transfusion by performing univariate analysis, followed by logistic regression. We then simplified the score to an integer-based system and tested it using the area under the receiver operator characteristic (AUC) statistic with a Hosmer-Lemeshow goodness-of-fit test. Finally, the scoring system was applied to the external validation dataset and the same statistical methods applied to test the accuracy of the ACTA-PORT score. Several factors were independently associated with risk of transfusion, including age, sex, body surface area, logistic EuroSCORE, preoperative haemoglobin and creatinine, and type of surgery. In our primary dataset, the score accurately predicted risk of perioperative transfusion in cardiac surgery patients with an AUC of 0.76. The external validation confirmed accuracy of the scoring method with an AUC of 0.84 and good agreement across all scores, with a minor tendency to under-estimate transfusion risk in very high-risk patients. The ACTA-PORT score is a reliable, validated tool for predicting risk of transfusion for patients undergoing cardiac surgery. This and other scores can be used in research studies for risk adjustment when assessing outcomes, and might also be incorporated into a Patient Blood Management programme.


Promoting Antenatal Care Attendance Through a Text Messaging Intervention in Samoa: Quasi-Experimental Study.
Antenatal care (ANC) has the potential to improve maternal health, but it remains underutilized and unevenly implemented in many low- and middle-income countries. Increasingly, text messaging programs for pregnant women show evidence that they can improve the utilization of ANC during pregnancy; however, gaps remain regarding how implementation affects outcomes. This study aimed to assess facilitators and barriers to implementation of an SMS text messaging intervention for pregnant women in Samoa and to assess its impact on ANC attendance. This study took place in Upolu, Samoa, from March to August 2014 and employed a quasi-experimental design. Half (n=3) of the public antenatal clinics on the island offered adult pregnant women the SMS text messaging intervention, with 552 women registering for the messages. At the comparison clinics (n=3), 255 women registered and received usual care. The intervention consisted of unidirectional text messages containing health tips and appointment reminders. The outcome of interest was the number of attended antenatal visits. Implementation data were also collected through a survey of the participating midwives (n=7) and implementation notes. Data analysis included a comparison of women's baseline characteristics between the two groups, followed by the use of negative binomial regressions to test for associations between participation in the intervention and increased ANC attendance, controlling for individual characteristics and accounting for the clustering of women within clinics. The comparison of ANC attendance rates found that women receiving the SMS text messaging intervention attended 15% fewer ANC visits than the comparison group (P=.004), controlling for individual characteristics and clustering. Data analysis of the implementation process suggests that barriers to successful implementation include women registering very late in pregnancy, sharing their phone with others, and inconsistent explanation of the intervention to women. These results suggest that unidirectional text messages do not encourage, and might even discourage, ANC attendance in Samoa. Interpreted with other evidence in the literature, these results suggest that SMS text messaging interventions are more effective when they facilitate better communication between patients and health workers. This study is an important contribution to our understanding of when SMS text messaging interventions are and are not effective in improving maternal health care utilization.


Signal transduction by the CD2 antigen in T cells and natural killer cells: requirement for expression of a functional T cell receptor or binding of antibody Fc to the Fc receptor, Fc gamma RIIIA (CD16).
Crosslinking of CD2 antigen on T lymphocytes and natural killer (NK) cells leads to a rise in cytoplasmic-free Ca2+ concentration ([Ca2+]i). However, CD2 seems unlikely to interact directly with the second messenger pathways since signaling via CD2 is poor in T cells that lack the T cell receptor (TCR) and is absent in L cells or insect cells that express CD2. In contrast, NK cells that are also TCR- can be triggered via CD2, but it is unclear as to whether the CD16 Fc receptor (FcR) may facilitate this effect. The CD16 transmembrane molecule is expressed in a complex with the zeta homodimer or the zeta/gamma heterodimer and these dimers are also associated with the TCR complex. Thus, it seemed that zeta chains may provide the link between signaling on NK cells and T cells. This could be tested on TCR- cells since when CD16 is transfected into T cells it is expressed in a complex with TCR zeta homodimer or the zeta/gamma heterodimer. At first, potentiation of CD2 signaling was seen on TCR- Jurkat cells expressing CD16, but this was found to be dependent on trace levels (1%) of IgG in F(ab')2 antibody preparations. With pure F(ab')2, the effect was lost. Signaling on a rat NK cell line was also re-examined with F(ab')2 antibodies that had no IgG contamination, and again no signal transduction via CD2 was seen. We thus conclude that there is no clear evidence for potent signaling via CD2 on cells that lack a TCR complex and that TCR zeta chain expressed at the cell surface is not sufficient to potentiate signaling via CD2 as measured by an increase in [Ca2+]i.


Differences in self-rated health among older immigrants--a comparison between older Finland-Swedes and Finns in Sweden.
Research has shown a tendency among immigrants and ethnic minorities to have a lower health status compared with the majority population. This applies to Finnish immigrants in Sweden. This group, however, also consists of persons who belong to a small ethnic minority in Finland, the Finland-Swedes, who speak Swedish as their mother tongue. In Finland, this minority has been shown to have better health and longer lives than the majority of Finnish-speaking people. Most of the previous research has studied the objective health of immigrants and minorities, while less is known about their subjective health. The aim of this study was to describe and compare self-rated health in older Finland-Swedes and Finns living as immigrants in Sweden. The study was carried out as a sample-based cross-sectional study. Data was collected by a postal structured questionnaire. The response rate among the Finland-Swedes was 47% (n = 169) and among the Finns was 54% (n = 643). Data was analyzed descriptively and tested with Pearson's chi-square test. The results showed significant differences between the language groups in self-rated health, age of retirement and causes for retirement. The Finland-Swede immigrants rated their health as better than the Finnish-speaking ones. They had retired at an older age and less frequently because of health problems. The results indicate that there may be significant differences in health, at least in subjective health, between immigrant groups. Due to the low response rate, the results cannot be generalized.


Immunolocalization of androgen receptors and aromatase enzyme in the adult musk shrew brain.
In the brain and other tissues, estrogens are produced by aromatization of androgens. Biochemical data suggest that aromatase enzyme is regulated by the androgen receptor (AR). Neurons that contain either AR or aromatase (AROM) enzyme reside in many of the same brain regions. In this report, we examined the codistribution of AR- and AROM-enzyme-immunoreactive (-ir) neurons in several regions of the adult male and female musk shrew brain. Data were collected from the intermediate nucleus of the lateral septum (LS), medial anterior (BNSTMA) and medial posterointerior (BNSTMP) divisions of the bed nucleus of the stria terminalis, medial preoptic area (mPOA), ventromedial nucleus of the hypothalamus (VMN), medial (MeA), cortical and central nuclei of the amygdala. Males had significantly more AR-ir neurons in the BNSTMP, mPOA, VMN and LS as compared to females. With the exception of the BNSTMA and LS, males had more AROM-ir neurons in each region than females. Furthermore, males had significantly more double-labeled neurons than females in the BNSTMP, mPOA, VMN, LS and MeA. The percentage of AROM-ir neurons that also contained AR immunoreactivity ranged from 13 to 82% depending on sex and region. The highest percentage of dual-labeled neurons (79% in females and 82% in males) was found in the VMN. Taken together, these data show that there is extensive cellular colocalization of AR and AROM enzyme in specific regions of the musk shrew brain. We propose that in both sexes, androgen receptors may act as transcription factors to regulate AROM enzyme.


The C2H2-type transcription factor, FlbC, is involved in the transcriptional regulation of Aspergillus oryzae glucoamylase and protease genes specifically expressed in solid-state culture.
Aspergillus oryzae produces a large amount of secreted proteins in solid-state culture, and some proteins such as glucoamylase (GlaB) and acid protease (PepA) are specifically produced in solid-state culture, but rarely in submerged culture. From the disruption mutant library of A. oryzae transcriptional regulators, we successfully identified a disruption mutant showing an extremely low production level of GlaB but a normal level of α-amylase production. This strain was a disruption mutant of the C2H2-type transcription factor, FlbC, which is reported to be involved in the regulation of conidiospore development. Disruption mutants of other upstream regulators comprising a conidiation regulatory network had no apparent effect on GlaB production in solid-state culture. In addition to GlaB, the production of acid protease in solid-state culture was also markedly decreased by flbC disruption. Northern blot analyses revealed that transcripts of glaB and pepA were significantly decreased in the flbC disruption strain. These results suggested that FlbC is involved in the transcriptional regulation of genes specifically expressed under solid-state cultivation conditions, possibly independent of the conidiation regulatory network.


Targeting of inflammatory pathways with R2CHOP in high-risk DLBCL.
Diffuse large B-cell lymphoma (DLBCL) is the most common lymphoma, and front line therapies have not improved overall outcomes since the advent of immunochemotherapy. By pairing DNA and gene expression data with clinical response data, we identified a high-risk subset of non-GCB DLBCL patients characterized by genomic alterations and expression signatures capable of sustaining an inflammatory environment. These mutational alterations (PIM1, SPEN, and MYD88 [L265P]) and expression signatures (NF-κB, IRF4, and JAK-STAT engagement) were associated with proliferative signaling, and were found to be enriched in patients treated with RCHOP that experienced unfavorable outcomes. However, patients with these high-risk mutations had more favorable outcomes when the immunomodulatory agent lenalidomide was added to RCHOP (R2CHOP). We are the first to report the genomic validation of a high-risk phenotype with a preferential response towards R2CHOP therapy in non-GCB DLBCL patients. These conclusions could be translated to a clinical setting to identify the ~38% of non-GCB patients that could be considered high-risk, and would benefit from alternative therapies to standard RCHOP based on personalized genomic data.


Purification and characterization of the plasmodial phosphatase that hydrolyses the phosphorylated light chain of Physarum myosin II from Physarum polycephalum.
A phosphatase was purified through a combination of ion-exchange and hydrophobic chromatography followed by native PAGE from Physarum plasmodia. Recently, we demonstrated that this phosphatase isoform has a hydrolytic activity towards the PMLC (phosphorylated light chain of Physarum myosin II) at pH 7.6. The apparent molecular mass of the purified enzyme was estimated at approximately 50 kDa by means of analytical gel filtration. The enzyme was purified 340-fold to a final phosphatase activity of 400 pkat/mg of protein. Among the phosphorylated compounds tested for hydrolytic activity at pH 7.6, the enzyme showed no activity towards nucleotides. At pH 7.6, hydrolytic activity of the enzyme against PMLC was detected; at pH 5.0, however, no hydrolytic activity towards PMLC was observed. The Km of the enzyme for PMLC was 10 microM, and the V(max) was 1.17 nkat/mg of protein. Ca(2+) (10 microM) inhibited the activity of the enzyme, and Mg(2+) (8.5 microM) activated the dephosphorylation of PMLC. Mn(2+) (1.6 microM) highly stimulated the enzyme's activity. Based on these results, we concluded that the enzyme is likely to be a phosphatase with hydrolytic activity towards PMLC.


Edge effects in 3D dosimetry: characterisation and correction of the non-uniform dose response of PRESAGE®.
Previous work has shown that PRESAGE® can be used successfully to perform 3D dosimetric measurements of complex radiotherapy treatments. However, measurements near the sample edges are known to be difficult to achieve. This is an issue when the doses at air-material interfaces are of interest, for example when investigating the electron return effect (ERE) present in treatments delivered by magnetic resonance (MR)-linac systems. To study this effect, a set of 3.5 cm-diameter cylindrical PRESAGE® samples was uniformly irradiated with multiple dose fractions, using either a conventional linac or an MR-linac. The samples were imaged between fractions using an optical-CT, to read out the corresponding accumulated doses. A calibration between TPS-predicted dose and optical-CT pixel value was determined for individual dosimeters as a function of radial distance from the axis of rotation. This data was used to develop a correction that was applied to four additional samples of PRESAGE® of the same formulation, irradiated with 3D-CRT and IMRT treatment plans, to recover significantly improved 3D measurements of dose. An alternative strategy was also tested, in which the outer surface of the sample was physically removed prior to irradiation. Results show that for the formulation studied here, PRESAGE® samples have a central region that responds uniformly and an edge region of 6-7 mm where there is gradual increase in dosimeter response, rising to an over-response of 24%-36% at the outer boundary. This non-uniform dose response increases in both extent and magnitude over time. Both mitigation strategies investigated were successful. In our four exemplar studies, we show how discrepancies at edges are reduced from 13%-37% of the maximum dose to between 2 and 8%. Quantitative analysis shows that the 3D gamma passing rates rise from 90.4, 69.3, 63.7 and 43.6% to 97.3, 99.9, 96.7 and 98.9% respectively.


Genetic heterogeneity and clinical variability in musculocontractural Ehlers-Danlos syndrome caused by impaired dermatan sulfate biosynthesis.
Bi-allelic variants in CHST14, encoding dermatan 4-O-sulfotransferase-1 (D4ST1), cause musculocontractural Ehlers-Danlos syndrome (MC-EDS), a recessive disorder characterized by connective tissue fragility, craniofacial abnormalities, congenital contractures, and developmental anomalies. Recently, the identification of bi-allelic variants in DSE, encoding dermatan sulfate epimerase-1 (DS-epi1), in a child with MC-EDS features, suggested locus heterogeneity for this condition. DS-epi1 and D4ST1 are crucial for biosynthesis of dermatan sulfate (DS) moieties in the hybrid chondroitin sulfate (CS)/DS glycosaminoglycans (GAGs). Here, we report four novel families with severe MC-EDS caused by unique homozygous CHST14 variants and the second family with a homozygous DSE missense variant, presenting a somewhat milder MC-EDS phenotype. The glycanation of the dermal DS proteoglycan decorin is impaired in fibroblasts from D4ST1- as well as DS-epi1-deficient patients. However, in D4ST1-deficiency, the decorin GAG is completely replaced by CS, whereas in DS-epi1-deficiency, still some DS moieties are present. The multisystemic abnormalities observed in our patients support a tight spatiotemporal control of the balance between CS and DS, which is crucial for multiple processes including cell differentiation, organ development, cell migration, coagulation, and connective tissue integrity.


Comparison of mouse and human keratin 18: a component of intermediate filaments expressed prior to implantation.
Keratin 18 is a type-I keratin that is found in a variety of simple epithelial tissues. In mice, the corresponding protein, called Endo B, is expressed at the 4- to 8-cell stage of mouse development and may be one of the first intermediate-filament proteins synthesized after fertilization. A cDNA clone for keratin 18, designated pK18, was isolated from a human placental cDNA library by hybridization with the mouse Endo-B probe. It was characterized by hybridization selection of RNA, translation, immunoprecipitation, Northern blotting, and sequence analysis. Synthetic T7 polymerase transcripts of the cDNA were indistinguishable in size from keratin-18 mRNA, suggesting that pK18 represents a full-length copy of the RNA. The cDNA insert is 1,428 nucleotides long and contains a single open reading frame of 1,342 nucleotides coding for 429 amino acids. The deduced amino acid sequence is 89.7% identical with that of Endo B. The only extensive difference between the two sequences is due to 9 additional amino acids being present in the last half of the N-terminal domain of keratin 18. The 38-nucleotide-long 3' noncoding region of the cDNA is 75% identical with the corresponding portion of Endo B. The 5' noncoding regions are 59% identical. The expression of keratin-18 mRNA was found to vary more than tenfold when HeLa cells and BeWo trophoblastic cells were compared.


Sex Differences in Psychiatric Hospitalizations of Individuals With Psychotic Disorders.
We aimed to evaluate the association between sex and hospitalization characteristics in psychotic disorders. We identified all acute hospitalizations, between 2010 and 2013, for psychotic disorders in patients younger than 45 and older than 55 years (n = 5411) in the hospital's database. In addition, we identified patients who were prescribed with intramuscular risperidone (n = 280) or clozapine (n = 192) at discharge. The results showed that women younger than 45 years had lower proportions of hospitalizations (33.52% vs. 66.47%) and involuntary hospitalizations (33.85% vs. 45.55%) than did men in the same age group. Women older than 55 years had higher proportions of hospitalizations (57.22% vs. 42.77%) and similar proportion of involuntary hospitalizations. Women younger than 45 years were prescribed similar doses of intramuscular risperidone and lower doses of clozapine (345.8 vs. 380.2 mg) and women older than 55 years were prescribed higher doses of intramuscular risperidone (44.8 vs. 34.4 mg/2 weeks) and clozapine (164.32 vs. 154.5 mg) than were men in the same age group. Women in the reproductive years have better hospitalization characteristics than do men on these measures.


belg: A Tool for Calculating Boltzmann Entropy of Landscape Gradients.
Entropy is a fundamental concept in thermodynamics that is important in many fields, including image processing, neurobiology, urban planning, and sustainability. As of recently, the application of Boltzmann entropy for landscape patterns was mostly limited to the conceptual discussion. However, in the last several years, a number of methods for calculating Boltzmann entropy for landscape mosaics and gradients were proposed. We developed an R package belg as an open source tool for calculating Boltzmann entropy of landscape gradients. The package contains functions to calculate relative and absolute Boltzmann entropy using the hierarchy-based and the aggregation-based methods. It also supports input raster with missing (NA) values, allowing for calculations on real data. In this study, we explain ideas behind implemented methods, describe the core functionality of the software, and present three examples of its use. The examples show the basic functions in this package, how to adjust Boltzmann entropy values for data with missing values, and how to use the belg package in larger workflows. We expect that the belg package will be a useful tool in the discussion of using entropy for a description of landscape patterns and facilitate a thermodynamic understanding of landscape dynamics.


Regulatory peptides of the gut and brain.
The concept of the gut-brain axis has emerged from the recent demonstration, mainly by radioimmunoassay and immunohistochemistry, that many peptides are common to both gut and brain cells. The general term regulatory peptide is at present the most appropriate for these peptides until more is known about their precise hormonal, neurotransmitter and neuromodulator activities. Consideration of possible local endocrine (paracrine) and neuroendocrine secretion necessitates re-evaluation of the importance of peripheral regulatory peptide concentrations. Central and peripheral administration of these peptides is a useful way to establish their potential regulatory roles in endocrine and nervous pathways. The concept of the gut-brain axis demonstrates the intimate relations between endocrinology and neuroendocrinology.


Genetics of Meesmann corneal dystrophy: a novel mutation in the keratin 3 gene in an asymptomatic family suggests genotype-phenotype correlation.
Juvenile epithelial corneal dystrophy of Meesmann (MCD, OMIM 122100) is a dominantly inherited disorder characterized by fragility of the anterior corneal epithelium and intraepithelial microcyst formation. Although the disease is generally mild and affected individuals are often asymptomatic, some suffer from recurrent erosions leading to lacrimation, photophobia, and deterioration in visual acuity. MCD is caused by mutations in keratin 3 (KRT3) or keratin 12 (KRT12) genes, which encode cornea-specific cytoskeletal proteins. Seventeen mutations in KRT12 and two in KRT3 have been described so far. The purpose of this study was to investigate the genetic background of MCD in a Polish family. We report on a three-generation family with MCD. Epithelial lesions characteristic for MCD were visualized with slit-lamp examination and confirmed by in vivo confocal microscopy. Using genomic DNA as a template, all coding regions of KRT3 and KRT12 were amplified and sequenced. Presence of the mutation was verified with restriction endonuclease digestion. In the proband, direct sequencing of the polymerase chain reaction (PCR) product from amplified coding regions of KRT3 and KRT12 revealed a novel 1493A>T heterozygous missense mutation in exon 7 of KRT3, which predicts the substitution of glutamic acid for valine at codon 498 (E498V). Using PCR-Restriction Fragment Length Polymorphism (RFLP) analysis, the mutation was demonstrated to segregate with the disease (four affected members, three non-affected) and to be absent in 100 controls from the Polish population, indicating that it is not a common polymorphism. Location of the E498V mutation emphasizes the functional relevance of the highly conserved boundary motifs at the COOH-terminus of the alpha-helical rod domain in keratin 3 (K3).


Group index and group velocity dispersion in silicon-on-insulator photonic wires.
We determine group index and group velocity dispersion (GVD) of SOI single-mode strip waveguides (photonic wires) with 525x226nm cross-section over the entire telecommunication bandwidth by employing an integrated Mach-Zehnder interferometer. The measured GVD yields 4400 ps/(nm*km) at 1550 nm and exceeds that of standard single-mode fibers by almost three orders of magnitude. In the photonic wires the GVD is mainly determined by strong light confinement rather than by material dispersion. Our results indicate that despite this high GVD, dispersion-induced signal impairment is negligible in photonic circuits for data rates up to 100-Gb/s and total waveguide lengths as long as about 1 meter. The measured group index and GVD are used as benchmarks to compare model calculations originating from four different theoretical methods.


Theory of photoionization-induced blueshift of ultrashort solitons in gas-filled hollow-core photonic crystal fibers.
We show theoretically that the photoionization process in a hollow-core photonic crystal fiber filled with a Raman-inactive noble gas leads to a constant acceleration of solitons in the time domain with a continuous shift to higher frequencies, limited only by ionization loss. This phenomenon is opposite to the well-known Raman self-frequency redshift of solitons in solid-core glass fibers. We also predict the existence of unconventional long-range nonlocal soliton interactions leading to spectral and temporal soliton clustering. Furthermore, if the core is filled with a Raman-active molecular gas, spectral transformations between redshifted, blueshifted, and stabilized solitons can take place in the same fiber.


Coping strategies for antidepressant side effects: an Internet survey.
Patients' coping methods to palliate side effects of antidepressants have not been reported in the literature. Through an Internet survey, 856 participants who were diagnosed with depression and receiving antidepressants were recruited to report on the methods of coping with side effects. They were asked which side effect(s) they experienced and to write freely about the way they tried to counteract these effects. We classified active coping methods into the following sub-types: adjustment of prescriptions, additional medication, complementary therapy, consultation with physicians, and daily relief. The prevalence of active coping differed across side effects (from 26.7%, sexual dysfunction, to 89.5%, dry mouth). Events with a lower percentage of active coping were more likely to be managed with "adjustment of prescriptions": (sexual dysfunction, 41.9%; fatigue, 36.8%; sweating, 20.0%; tremor, 42.5%; and somnolence, 31.8%). Further, a strong negative correlation was found between the percentage of participants reporting an adjustment of prescription and that reporting an active coping (r=-0.907, p<0.001). The "daily relief" sub-type contained a variety of strategies, including negative methods such as vomiting for nausea and weight gain and drinking alcohol for insomnia. Sampling of subjects were biased due to an Internet survey and diagnosis of depression and experience of side effects were self-reported. Patients with depression use various ways in alleviating antidepressants side effects. Some effects such as sexual dysfunction and fatigue may not be amenable to subjective coping efforts and others are sometimes managed inappropriately, which warrants a prudent attention.


Understanding the role of frustrated Lewis pairs as ligands in transition metal-catalyzed reactions.
The role of frustrated Lewis pairs (FLPs) as ligands in gold(i) catalyzed-reactions has been computationally investigated by using state-of-the-art density functional theory calculations. To this end, the nature of (P,B)-FLP-transition metal interactions in different gold(i)-complexes has been first explored in detail with the help of the energy decomposition analysis method, which allowed us to accurately quantify the so far poorly understood AuB interactions present in these species. The impact of such interactions on the catalytic activity of gold(i)-complexes has been then evaluated by performing the Au(i)-catalyzed hydroarylation reaction of phenylacetylene with mesitylene. With the help of the activation strain model of reactivity, the factors governing the higher activity of Au(i)-complexes having a FLP as a ligand as compared to that of the parent PPh3 system have also been quantitatively identified.


I Keep a Close Watch on This Heart of Mine: Increased Interoception in Insomnia.
Whereas both insomnia and altered interoception are core symptoms in affective disorders, their neural mechanisms remain insufficiently understood and have not previously been linked. Insomnia Disorder (ID) is characterized by sensory hypersensitivity during wakefulness and sleep. Previous studies on sensory processing in ID addressed external stimuli only, but not interoception. Interoceptive sensitivity can be studied quantitatively by measuring the cerebral cortical response to one's heartbeat (heartbeat-evoked potential, HEP). We here investigated whether insomnia is associated with increased interoceptive sensitivity as indexed by the HEP amplitude. Sixty-four participants aged 21-70 years were recruited through www.sleepregistry.nl including 32 people suffering from ID and 32 age- and sex-matched controls without sleep complaints. HEPs were obtained from resting-state high-density electroencephalography (HD-EEG) recorded during evening wakeful rest in eyes-open (EO) and eyes-closed (EC) conditions of 5-minute duration each. Significance of group differences in HEP amplitude and their topographical distribution over the scalp were assessed by means of cluster-based permutation tests. In particular during EC, and to a lesser extent during EO, people with ID had a larger amplitude late HEP component than controls at frontal electrodes 376-500 ms after the R-wave peak. Source localization suggested increased neural activity time-locked to heartbeats in people with ID mainly in anterior cingulate/medial frontal cortices. People with insomnia show insufficient adaptation of their brain responses to the ever-present heartbeats. Abnormalities in the neural circuits involved in interoceptive awareness including the salience network may be of key importance to the pathophysiology of insomnia.


[Inhibition of Bacillus coagulans growth in laboratory media and in fruit purees].
The growth of two strains of B. coagulans was inhibited in laboratory media at pH < or = 4.5, and at water activity (aw) levels of 0.96 for B. coagulans NRS 609 and 0.95 for B. coagulans ATCC 803. The growth of both strains was also inhibited in apple and strawberry purees (pH = 3.5) stored at 37 degrees C for over two months. B. coagulans was able to grow in banana puree (pH approximately equal to 5.0) but acidification of the puree at pH = 3.5 was enough to prevent growth. The addition of up to 3,000 ppm vainillin ("natural" preservative) or 1,000 ppm potassium sorbate (traditional preservative) at pH higher than the inhibitory level previously determined could not prevent growth of B. coagulans in laboratory or in fruits, but 100 ppm lysozyme retarded growth in laboratory media at different pH levels (from 4.5 to 6.7) and in banana puree. As lysozyme showed to be effective at pH < or = 6.7, it might be used to prevent growth of B. coagulans at an eventual increment of pH during storage.


Germ Cell Responses to Stress: The Role of RNP Granules.
The ability to respond to stress is critical to survival for animals. While stress responses have been studied at both organismal and cellular levels, less attention has been given to the effect of stress on the germ line. Effective germ line adaptations to stress are essential to the propagation of a species. Recent studies suggest that germ cells share some cellular responses to stress with somatic cells, including the assembly of RNP granules, but may also have unique requirements. One fundamental difference between oocytes and sperm, as well as most somatic cells, is the long lifespan of oocytes. Since women are born with all of their eggs, oocytes must maintain their cellular quality over decades prior to fertilization. This prolonged meiotic arrest is one type of stress that eventually contributes to decreased fertility in older women. Germ cell responses to nutritional stress and heat stress have also been well-characterized using model systems. Here we review our current understanding of how germ cells respond to stress, with an emphasis on the dynamic assembly of RNP granules that may be adaptive. We compare and contrast stress responses of male gametes with those of female gametes, and discuss how the dynamic cellular remodeling of the germ line can impact the regulation of gene expression. We also discuss the implications of recent in vitro studies of ribonucleoprotein granule assembly on our understanding of germ line responses to stress, and the gaps that remain in our understanding of the function of RNP granules during stress.


Nasal swab samples and real-time polymerase chain reaction assays in community-based, longitudinal studies of respiratory viruses: the importance of sample integrity and quality control.
Carefully conducted, community-based, longitudinal studies are required to gain further understanding of the nature and timing of respiratory viruses causing infections in the population. However, such studies pose unique challenges for field specimen collection, including as we have observed the appearance of mould in some nasal swab specimens. We therefore investigated the impact of sample collection quality and the presence of visible mould in samples upon respiratory virus detection by real-time polymerase chain reaction (PCR) assays. Anterior nasal swab samples were collected from infants participating in an ongoing community-based, longitudinal, dynamic birth cohort study. The samples were first collected from each infant shortly after birth and weekly thereafter. They were then mailed to the laboratory where they were catalogued, stored at -80°C and later screened by PCR for 17 respiratory viruses. The quality of specimen collection was assessed by screening for human deoxyribonucleic acid (DNA) using endogenous retrovirus 3 (ERV3). The impact of ERV3 load upon respiratory virus detection and the impact of visible mould observed in a subset of swabs reaching the laboratory upon both ERV3 loads and respiratory virus detection was determined. In total, 4933 nasal swabs were received in the laboratory. ERV3 load in nasal swabs was associated with respiratory virus detection. Reduced respiratory virus detection (odds ratio 0.35; 95% confidence interval 0.27-0.44) was observed in samples where the ERV3 could not be identified. Mould was associated with increased time of samples reaching the laboratory and reduced ERV3 loads and respiratory virus detection. Suboptimal sample collection and high levels of visible mould can impact negatively upon sample quality. Quality control measures, including monitoring human DNA loads using ERV3 as a marker for epithelial cell components in samples should be undertaken to optimize the validity of real-time PCR results for respiratory virus investigations in community-based studies.


Preparation and characterization of amphiphilic poly-N-vinylpyrrolidone nanoparticles containing indomethacin.
Amphiphilic poly-N-vinylpyrrolidone derivatives (Amph-PVP) with different molecular weight of hydrophilic PVP fragment and one terminal hydrophobic n-alkyl fragment of different length were synthesized for preparation of nano-scaled particles in aqueous media. To estimate novel polymer efficiency and perspective as basis for drug delivery systems, the polymeric micelle-like particles were prepared by dialysis and solvent evaporation methods. Indomethacin was incorporated into hydrophobic inner core of these nanoparticles as a typical model drug. From the dynamic light-scattering measurements, the size of particles formed was less than 200 nm with narrow monodisperse size distribution and nanoparticles size slightly increased with the amount of indomethacin encapsulated into inner core of Amph-PVP particles. The critical aggregation concentration values for prepared polymer samples determined by fluorescence spectroscopy were in micromole range which is lower than it is for common low molecular weight surfactants. As the hydrophobic fragment of amphiphilic polymer increased, the critical aggregation concentration values decreased. An increase of polymer hydrophilic fragment molecular weight produced larger nanoaggregates. In vitro release experiments using indomethacin-loaded Amph-PVP nanoparticles exhibited the sustained release behavior without any burst effect for most polymer samples.


[Prevalence and cervical human papilloma virus associated factors in patients with rheumatoid arthritis].
Nevertheless its association with cervicouterine cancer, there is no information about cervical human papillomavirus infection prevalence in patients with rheumatoid arthritis. To evaluate human papillomavirus infection prevalence through molecular biology tests, and to analyze this infection related factors in patients with rheumatoid arthritis. Analytic, transversal study to 250 patients: 61 women with rheumatoid arthritis selected from a rheumatologic external consult of a second level hospital, and 189 healthy women, with cervical cytology, of a first level hospital. They were polled to find infection risk factors. They were exfoliated to get cervix cells to extract its DNA and detect human papillomavirus (chain reaction of polymerase with specific consensus markers), and identification of restriction enzyme in high and low risks viruses. Prevalence was calculated, and adjusted factors analysis was performed through logistic regression with odds ratio and confidence intervals of 95%. Prevalence of papillomavirus infection in patients with rheumatoid arthritis was 30%, and in control group was 24%, with an odds ratio of 0.8 (CI 95% 0.42-1.6, p = 0.5). Ninety-four percent of the most frequent viral types in women with rheumatoid arthritis were high risk (mainly types 16, 58, and 18). Factors associated with higher human papillomavirus adjusted to rheumatoid arthritis were: more than one sexual partner (OR = 5.8 CI 95% 1.1-31.1, p = 0.04), more than one sexual intercourse weekly (OR = 6.7, CI 95% 0.9-51.6, p = 0.06), circumcised sexual partner (OR = 9.0, CI 95% 1.2-64.4, p = 0.02). Patients and controls had same values of marital status. Seventy-four percent of controls worked, compared to 44% of women with rheumatoid arthritis (p < 0.01). One out of three women with rheumatoid arthritis has human papillomavirus infection and 94% has the high-risk viral type. Infection associated factors mainly includes sexual partner ones; due to high risk of cervical dysplasia, it is necessary the early detection of the infection and surveillance.


Molecular underpinnings of ssDNA specificity by Rep HUH-endonucleases and implications for HUH-tag multiplexing and engineering.
Replication initiator proteins (Reps) from the HUH-endonuclease superfamily process specific single-stranded DNA (ssDNA) sequences to initiate rolling circle/hairpin replication in viruses, such as crop ravaging geminiviruses and human disease causing parvoviruses. In biotechnology contexts, Reps are the basis for HUH-tag bioconjugation and a critical adeno-associated virus genome integration tool. We solved the first co-crystal structures of Reps complexed to ssDNA, revealing a key motif for conferring sequence specificity and for anchoring a bent DNA architecture. In combination, we developed a deep sequencing cleavage assay, termed HUH-seq, to interrogate subtleties in Rep specificity and demonstrate how differences can be exploited for multiplexed HUH-tagging. Together, our insights allowed engineering of only four amino acids in a Rep chimera to predictably alter sequence specificity. These results have important implications for modulating viral infections, developing Rep-based genomic integration tools, and enabling massively parallel HUH-tag barcoding and bioconjugation applications.


Digital image registration using projections.
In many application fields (e.g., aerospace and biomedical image processing), one has to deal with a sequence of images whose observation is made difficult by unpredictable relative movements of the camera and the scene. In order to obtain a stable display, the displacement of each image in the sequence with respect to one chosen as a reference must be preliminarily determined. This would usually require two-dimensional algorithms, involving a considerable computational effort. This work discusses a new algorithm for image registration, which requires only one-dimensional Fourier transformations. Preliminary experimental results are reported.


Early life exposures contributing to accelerated lung function decline in adulthood - a follow-up study of 11,000 adults from the general population.
We aimed to assess whether exposure to risk factors in early life from conception to puberty continue to contribute to lung function decline later in life by using a pooled cohort comprising approx. 11,000 adults followed for more than 20 years and with up to three lung function measurements. Participants (20-68 years) in the ECRHS and NFBC1966 cohort studies followed in the periods 1991-2013 and 1997-2013, respectively, were included. Mean annual decline in maximum forced expired volume in 1 s (FEV1) and forced vital capacity (FVC) were main outcomes. Associations between early life risk factors and change in lung function were estimated using mixed effects linear models adjusted for sex, age, FEV1, FVC and height at baseline, accounting for personal smoking. Decline in lung function was accelerated in participants with mothers that smoked during pregnancy (FEV1 2.3 ml/year; 95% CI: 0.7, 3.8) (FVC 2.2 ml/year; 0.2, 4.2), with asthmatic mothers (FEV1 2.6 ml/year; 0.9, 4.4) (FEV1/FVC 0.04 per year; 0.04, 0.7) and asthmatic fathers (FVC 2.7 ml/year; 0.5, 5.0), and in women with early menarche (FVC 2.4 ml/year; 0.4, 4.4). Personal smoking of 10 pack-years contributed to a decline of 2.1 ml/year for FEV1 (1.8, 2.4) and 1.7 ml/year for FVC (1.3, 2.1). Severe respiratory infections in early childhood were associated with accelerated decline among ever-smokers. No effect-modification by personal smoking, asthma symptoms, sex or cohort was found. Mothers' smoking during pregnancy, parental asthma and early menarche may contribute to a decline of FEV1 and FVC later in life comparable to smoking 10 pack-years. European Union's Horizon 2020; Research Council of Norway; Academy of Finland; University Hospital Oulu; European Regional Development Fund; Spanish Ministry of Science and Innovation; Generalitat de Catalunya.


Mitogenic effect of muscle on the neuroepithelium of the developing spinal cord.
A previous study revealed that segments of bowel grafted between the neural tube and somites of a younger chick host embryo would induce a unilateral increase in cellularity of the host's neural tube. The current experiments were done to test the hypotheses that muscle tissue in the wall of the gut is responsible for this growth-promoting effect and that the spinal cord enlargement is the result of a mitogenic action on the neuroepithelium. Fragments of skeletal (E8-15) or cardiac muscle (E4-14) were removed from quail embryos and grafted between the neural tube and somites of chick host embryos (E2). Both skeletal and cardiac muscle grafts mimicked the effect of bowel and induced an increase in cell number as well as a unilateral enlargement of the region of the host's neural tube immediately adjacent to the grafts. The growth-promoting effect of muscle-containing grafts was restricted to the neural tube itself and was not seen in proximate dorsal root or sympathetic ganglia. The action of the grafts of muscle was neither species- nor class-specific, since enlargement of the neural tube was observed following implantation of fetal mouse skeletal muscle into quail hosts. Grafts of skeletal muscle or gut increased the number of cells taking up [3H]thymidine in the host's neuroepithelium as early as 9 h following implantation of a graft. The increase in the number of cells entering the S phase of the cell cycle preceded the increase in cell number. These observations demonstrate that muscle-containing tissues can increase the rate of proliferation of neuroepithelial cells when these tissues are experimentally placed together.


Normal and impaired reflexive orienting of attention after central nonpredictive cues.
Recent studies suggest that stimuli with directional meaning can trigger lateral shifts of visuospatial attention when centrally presented as noninformative cues. We investigated covert orienting in healthy participants and in a group of 17 right brain-damaged patients (9 with hemispatial neglect) comparing arrows, eye gaze, and digits as central nonpredictive cues in a detection task. Orienting effects elicited by arrows and eye gaze were overall consistent in healthy participants and in right brain-damaged patients, whereas digit cues were ineffective. Moreover, patients with neglect showed, at the shortest delay between cue and target, a disengage deficit for arrow cueing whose magnitude was predicted by neglect severity. We conclude that the peculiar form of attentional orienting triggered by the directional meaning of arrow cues presents some features previously thought to characterize only the stimulus-driven (exogenous) orienting to noninformative peripheral cues.


Cooperative light-induced breathing of soft porous crystals via azobenzene buckling.
Although light is a prominent stimulus for smart materials, the application of photoswitches as light-responsive triggers for phase transitions of porous materials remains poorly explored. Here we incorporate an azobenzene photoswitch in the backbone of a metal-organic framework producing light-induced structural contraction of the porous network in parallel to gas adsorption. Light-stimulation enables non-invasive spatiotemporal control over the mechanical properties of the framework, which ultimately leads to pore contraction and subsequent guest release via negative gas adsorption. The complex mechanism of light-gated breathing is established by a series of in situ diffraction and spectroscopic experiments, supported by quantum mechanical and molecular dynamic simulations. Unexpectedly, this study identifies a novel light-induced deformation mechanism of constrained azobenzene photoswitches relevant to the future design of light-responsive materials.


Direct-write polymer nanolithography in ultra-high vacuum.
Polymer nanostructures were directly written onto substrates in ultra-high vacuum. The polymer ink was coated onto atomic force microscope (AFM) probes that could be heated to control the ink viscosity. Then, the ink-coated probes were placed into an ultra-high vacuum (UHV) AFM and used to write polymer nanostructures on surfaces, including surfaces cleaned in UHV. Controlling the writing speed of the tip enabled the control over the number of monolayers of the polymer ink deposited on the surface from a single to tens of monolayers, with higher writing speeds generating thinner polymer nanostructures. Deposition onto silicon oxide-terminated substrates led to polymer chains standing upright on the surface, whereas deposition onto vacuum reconstructed silicon yielded polymer chains aligned along the surface.


Recognition and management of acute suppurative parotitis.
Acute suppurative parotitis (ASP) is a rare infectious process occurring after surgery or in debilitated patients. This case describes a 41-year-old man who was hospitalized initially for acute bacterial endocarditis. Late in the hospital course his recovery was complicated by the development of ASP. Subsequent cultures of blood and parotid-fluid samples grew Pseudomonas aeruginosa. Despite early empiric antibiotic therapy followed by more specific therapy with an aminoglycoside, the patient continued to experience severe pain, swelling, and facial-nerve palsy. Surgical incision and drainage of the parotid gland was accomplished on the ninth day after onset of ASP, and the patient experienced a rapid resolution of the disease process. Acute suppurative parotitis was originally attributed to infection with gram-positive organisms, primarily Staphylococcus aureus and Streptococcus species. More recently, reports of ASP caused by gram-negative and anaerobic bacteria have been published. This review describes the etiology, pathogenesis, microbiology, and clinical features of ASP. Nonpharmacologic, pharmacologic, and surgical management of the disease is discussed. Although ASP is a rare occurrence in the hospital setting, early recognition and appropriate management are important in avoiding serious morbidity and mortality.


Positive allosteric modulation of A1 adenosine receptors as a novel and promising therapeutic strategy for anxiety.
Activation of A1 adenosine receptors (ARs) has been associated with anxiolytic-like effects in different behavioral tests, but development of A1AR agonists for therapeutic use has been hampered, most likely due to the presence of side effects. With the aim to identify a safer approach for the treatment of anxiety, we investigated, in mice, the anxiolytic-like properties of a novel A1AR positive allosteric modulator, TRR469. Acute administration of TRR469 (0.3-3 mg/kg) resulted in robust anxiolytic-like effects in the elevated plus maze, the dark/light box, the open field and the marble burying tests. The magnitude of the anxiolytic action of TRR469 was comparable to that obtained with benzodiazepine diazepam (1 mg/kg). The use of the A1AR antagonist DPCPX (3 mg/kg) suggested that the effects of TRR469 were mediated by this receptor subtype. In contrast to diazepam, the novel positive allosteric modulator did not potentiate the sedative effect of ethanol (3.5 g/kg) evaluated by the loss of righting reflex. While diazepam produced motor coordination impairment in the rotarod test, this effect being enhanced by the presence of ethanol (1.5 g/kg), TRR469 did not elicit locomotor disturbances either when administered alone or in the presence of ethanol. In vitro, TRR469 was able to increase the number of A1AR recognizable by the agonist radioligand [3H]-CCPA in mouse brain regions involved in emotional processes. TRR469 markedly increased the affinity of the agonist CCPA, suggesting the capability, in vivo, to increase the affinity of endogenous adenosine. Taken together, these findings indicate that the positive allosteric modulation of A1AR may represent a promising approach for the treatment of anxiety-related disorders.


Differences in human alpha-, beta- and delta-globin gene expression in monkey kidney cells.
We have compared the function of the human alpha-, beta- and delta-globin genes using various plasmid expression vectors derived from pBR322. Amplification of recombinants occurred after their introduction, by calcium-phosphate-mediated DNA transfer, into monkey kidney cells that constitutively produce T antigen (COS cells). The human alpha-globin gene promoter functioned independently, but the beta-globin gene promoter was nearly totally dependent on the enhancing activity of the 72 bp direct repeats from the SV40 genome. Furthermore, when the human alpha- and beta-globin genes were linked in the same vector, the alpha promoter was active but the beta promoter was not. Function of the delta-globin gene promoter also depended on the enhancer element. In vectors containing the 72 bp repeats and the beta- or delta-globin gene, the activity of the beta-globin gene was approximately 50 times greater than that of the delta-globin gene, approximating the ratio of beta and delta mRNA observed in normal human bone marrow cells.


Posttraumatic Stress Disorder, Adverse Childhood Events, and Buccal Cell Telomere Length in Elderly Swiss Former Indentured Child Laborers.
Posttraumatic stress disorder (PTSD) is associated with increased risk for age-related diseases and early mortality. Accelerated biological aging could contribute to this elevated risk. The aim of the present study was to assess buccal cell telomere length (BTL) - a proposed marker of biological age - in men and women with and without PTSD. The role of childhood trauma was assessed as a potential additional risk factor for shorter telomere length. The sample included 62 former indentured Swiss child laborers (age: M = 76.19, SD = 6.18) and 58 healthy controls (age: M = 71.85, SD = 5.97). Structured clinical interviews were conducted to screen for PTSD and other psychiatric disorders. The Childhood Trauma Questionnaire (CTQ) was used to assess childhood trauma exposure. Quantitative polymerase chain reaction was used to measure BTL. Covariates include age, sex, years of education, self-evaluated financial situation, depression, and mental and physical functioning. Forty-eight (77.42%) of the former indentured child laborers screened positive for childhood trauma, and 21 (33.87%) had partial or full-blown PTSD. Results did not support our hypotheses that PTSD and childhood trauma would be associated with shorter BTL. In fact, results revealed a trend toward longer BTL in participants with partial or full PTSD [F(2,109) = 3.27, p = 0.04, η(2) = 0.06], and longer BTL was marginally associated with higher CTQ scores (age adjusted: β = 0.17 [95% CI: -0.01 to 0.35], t = 1.90, p = 0.06). Furthermore, within-group analyses indicated no significant association between BTL and CTQ scores. To the best of our knowledge, this is the first study exploring the association between childhood trauma and BTL in older individuals with and without PTSD. Contrary to predictions, there were no significant differences in BTL between participants with and without PTSD in our adjusted analyses, and childhood adversity was not associated with BTL. Possible explanations and future research possibilities are discussed.


Factors associated with ADL and IADL dependency among Korean centenarians: reaching the 100-year-old life transition.
This study aims to examine the functional status of Korean centenarians, who have crossed the relatively rare but increasingly common life transition of living for a century. As functional health is one of the essential components of healthy aging, our primary objective is to identify the correlates of two aspects of functioning, activities of daily living (ADL) and instrumental activities of daily living (IADL). Using a census survey conducted by the Korean National Statistical Office (KNSO) in 2005, we documented ADL and IADL limitations and analyzed their relationship to sociodemographic factors, health behaviors, social connectedness, and presence of diseases. The study participants were 796 adults aged 100 years or older. The mean age was 101.5 +/- 1.8, and females were 89.9% of the sample. Both ADL and IADL dependences were more common among females, those who did less physical activity, those who had more diseases, and those who did not participate in social activities. Intervention programs designed to address life-stage issues such as focusing on initiating healthy behaviors from youth, managing chronic diseases in mid-life, and fostering social participation in later life are recommended ways to improve functional independence and promote healthy aging among current and future generations of Korean centenarians.


Current strategies in nitrite detection and their application to field analysis.
The various analytical protocols that have been developed to aid the measurement of nitrite within environmental samples have been critically appraised and their applicability to field measurement assessed. The present communication presents a perspective on current techniques and technologies encompassing spectroscopic, electrochemical and chromatographic methodologies and highlights those that are liable to emerge in the near future. Commercial devices have been included and where appropriate the advantages and limitations posed by their operation within field contexts have been described.


Mutational and functional analysis in human Ras/MAP kinase genetic syndromes.
The Ras/mitogen-activated protein kinase (MAPK) pathway is essential in regulation of the cell cycle, cell differentiation, growth, and cell senescence, each of which are critical to normal development. A class of developmental disorders, the "RASopathies," is caused by germline mutations in genes that encode protein components of the Ras/MAPK pathway which result in dysregulation of the pathway and profound deleterious effects on development. One of these syndromes, cardiofaciocutaneous (CFC) syndrome, is caused by germline mutations in BRAF, MAP2K1 (MEK1) and MAP2K2 (MEK2), and possibly KRAS genes. Here, we describe the laboratory protocols and methods that we used to identify mutations in BRAF and MEK1/2 genes as causative for CFC syndrome. In addition, we present the techniques used to determine the effect these mutations have on activity of the Ras/MAPK pathway through Western blot analysis of the phosphorylation of endogenous ERK1/2, as well as through the use of an in vitro kinase assay that measures the phosphorylation of Elk-1.


Early life stress aggravates disease pathogenesis in mice with experimental autoimmune encephalomyelitis: Support for a two-hit hypothesis of multiple sclerosis etiology.
Vision problems are one of the earliest diagnosed symptoms of multiple sclerosis (MS). The onset and progression of vision loss and the underlying pathogenesis in MS may be influenced by cumulative psychophysiological stress. Here, we used a two-hit model of stress in female mice to determine if early life stress (ELS, the first hit) influences the response to an immunization that induces experimental autoimmune encephalomyelitis (EAE, the second hit) later in life. We hypothesized that ELS caused by animal transportation from a vendor during early postnatal development represents a co-factor which can exacerbate the clinical severity of EAE. Indeed, adult EAE mice with a history of ELS displayed more severe clinical signs and delayed recovery compared to non-stressed EAE mice. ELS also diminished visual acuity measured by optokinetic responses, as well as locomotion and exploratory behaviours in EAE mice. Notably, ELS accelerated vision loss and caused earlier onset of visual impairments in EAE. Exacerbated functional impairments in stressed EAE mice were highly correlated with circulating corticosterone levels. The findings show that the progression of induced EAE in adulthood can be significantly impacted by adverse early life experiences. These observations emphasize the importance of comprehensive behavioural testing, including non-motor functions, to enhance the translational value of preclinical animal models of MS. Moreover, shipment stress of laboratory animals should be considered a necessary variable in preclinical MS research. The consideration of cumulative lifetime stresses provides a new perspective of MS pathogenesis within a personalized medicine framework.


Computational fluid dynamics study on the influence of an alternate ventilation configuration on the possible flow path of infectious cough aerosols in a mock airborne infection isolation room.
When infectious epidemics occur, they can be perpetuated within health care settings, potentially resulting in severe health care workforce absenteeism, morbidity, mortality, and economic losses. The ventilation system configuration of an airborne infection isolation room is one factor that can play a role in protecting health care workers from infectious patient bioaerosols. Though commonly associated with airborne infectious diseases, the airborne infection isolation room design can also impact other transmission routes such as short-range airborne as well as fomite and contact transmission routes that are impacted by contagion concentration and recirculation. This article presents a computational fluid dynamics study on the influence of the ventilation configuration on the possible flow path of bioaerosol dispersal behavior in a mock airborne infection isolation room. At first, a mock airborne infection isolation room was modeled that has the room geometry and layout, ventilation parameters, and pressurization corresponding to that of a traditional ceiling-mounted ventilation arrangement observed in existing hospitals. An alternate ventilation configuration was then modeled to retain the linear supply diffuser in the original mock airborne infection isolation room but interchanging the square supply and exhaust locations to place the exhaust closer to the patient source and allow clean air from supply vents to flow in clean-to-dirty flow paths, originating in uncontaminated parts of the room prior to entering the contaminated patient's air space. The modeled alternate airborne infection isolation room ventilation rate was 12 air changes per hour. Two human breathing models were used to simulate a source patient and a receiving health care worker. A patient cough cycle was introduced into the simulation, and the airborne infection dispersal was tracked in time using a multi-phase flow simulation approach. The results from the alternate configuration revealed that the cough aerosols were pulled by the exhaust vent without encountering the health care worker by 0.93 s after patient coughs and the particles were controlled as the aerosols' flow path was uninterrupted by an air particle streamline from patient to the ceiling exhaust venting out cough aerosols. However, not all the aerosols were vented out of the room. The remaining cough aerosols entered the health care worker's breathing zone by 0.98 s. This resulted in one of the critical stages in terms of the health care worker's exposure to airborne virus and presented the opportunity for the health care worker to suffer adverse health effects from the inhalation of cough aerosols. Within 2 s, the cough aerosols reentered and recirculated within the patient and health care worker's surroundings resulting in pockets of old contaminated air. By this time, coalescence losses decreased as the aerosol were no longer in very close proximity and their movement was primarily influenced by the airborne infection isolation room airflow patterns. In the patient and health care worker's area away from the supply, the fresh air supply failed to reach this part of the room to quickly dilute the cough aerosol concentration. The exhaust was also found to have minimal effect upon cough aerosol removal, except for those areas with high exhaust velocities, very close to the exhaust grill. Within 5-20 s after a patient's cough, the aerosols tended to break up to form smaller sized aerosols of less than one micron diameter. They remained airborne and entrained back into the supply air stream, spreading into the entire room. The suspended aerosols resulted in the floating time of more than 21 s in the room due to one cough cycle. The duration of airborne contagion in the room and its prolonged exposure to the health care worker is likely to happen due to successive coughing cycles. Hence, the evaluated alternate airborne infection isolation room is not effective in removing at least 38% particles exposed to health care worker within the first second of a patient's cough.


Attraction of Lutzomyia longipalpis to synthetic sex-aggregation pheromone: Effect of release rate and proximity of adjacent pheromone sources.
In South America, the Protist parasite that causes visceral leishmaniasis, a potentially fatal human disease, is transmitted by blood-feeding female Lutzomyia longipalpis sand flies. A synthetic copy of the male produced sex-aggregation pheromone offers new opportunities for vector control applications. We have previously shown that the pheromone placed in plastic sachets (lures) can attract both females and males to insecticide treated sites for up to 3 months. To use the pheromone lure in a control program we need to understand how the application of lures in the field can be optimised. In this study we investigated the effect of increasing the number of lures and their proximity to each other on their ability to attract Lu. longipalpis. Also for the first time we applied a Bayesian log-linear model rather than a classic simple (deterministic) log-linear model to fully exploit the field-collected data. We found that sand fly response to pheromone is significantly related to the quantity of pheromone and is not influenced by the proximity of other pheromone sources. Thus sand flies are attracted to the pheromone source at a non-linear rate determined by the amount of pheromone being released. This rate is independent of the proximity of other pheromone releasing traps and indicates the role of the pheromone in aggregation formation. These results have important implications for optimisation of the pheromone as a vector control tool and indicate that multiple lures placed in relatively close proximity to each other (5 m apart) are unlikely to interfere with one another.


Hyperoxia toxicity in septic shock patients according to the Sepsis-3 criteria: a post hoc analysis of the HYPER2S trial.
Criteria for the Sepsis-3 definition of septic shock include vasopressor treatment to maintain a mean arterial pressure > 65 mmHg and a lactate concentration > 2 mmol/L. The impact of hyperoxia in patients with septic shock using these criteria is unknown. A post hoc analysis was performed of the HYPER2S trial assessing hyperoxia versus normoxia in septic patients requiring vasopressor therapy, in whom a plasma lactate value was available at study inclusion. Mortality was compared between patients fulfilling the Sepsis-3 septic shock criteria and patients requiring vasopressors for hypotension only (i.e., with lactate ≤ 2 mmol/L). Of the 434 patients enrolled, 397 had available data for lactate at inclusion. 230 had lactate > 2 mmol/L and 167 ≤ 2 mmol/L. Among patients with lactate > 2 mmol/L, 108 and 122 were "hyperoxia"- and "normoxia"-treated, respectively. Patients with lactate > 2 mmol/L had significantly less COPD more cirrhosis and required surgery more frequently. They also had higher illness severity (SOFA 10.6 ± 2.8 vs. 9.5 ± 2.5, p = 0.0001), required more renal replacement therapy (RRT), and received vasopressor and mechanical ventilation for longer time. Mortality rate at day 28 was higher in the "hyperoxia"-treated patients with lactate > 2 mmol/L as compared to "normoxia"-treated patients (57.4% vs. 44.3%, p = 0.054), despite similar RRT requirements as well as vasopressor and mechanical ventilation-free days. A multivariate analysis showed an independent association between hyperoxia and mortality at day 28 and 90. In patients with lactate ≤ 2 mmol/L, hyperoxia had no effect on mortality nor on other outcomes. Our results suggest that hyperoxia may be associated with a higher mortality rate in patients with septic shock using the Sepsis-3 criteria, but not in patients with hypotension alone.


Can fluoride-oxalate and sodium citrate stabilise homocysteine levels after blood collection?
The concentration of homocysteine (Hcy) rises rapidly after the collection of blood. This feature requires blood to be collected into the anticoagulants EDTA or heparin and the plasma to then be immediately separated; alternatively, the blood may be kept on ice and centrifuged within 1 hour. The use of chemical preservatives has been proposed as a means of stabilising Hcy levels in whole blood after collection. The objective of this study was to determine whether the commonly available fluoride-oxalate (Fl-Ox) and sodium citrate (Na-Cit) containers could stabilise Hcy levels in blood. Our results showed that when blood was collected into potassium EDTA (K-EDTA) tubes, Hcy levels rose from initial levels, on standing at room temperature (approximately 25 degrees C), by an average of 21% after 3 hours and 32% after 5 hours. The initial Hcy levels of blood collected into Fl-Ox and Na-Cit containers, however, were lower, at averages of 89% and 91%, respectively, compared to that of the same samples when collected into K-EDTA tubes. Hcy in these samples subsequently rose on standing, and after 5 hours was, on the average, 10 and 13% higher, respectively, compared with the initial levels in K-EDTA tubes. We conclude that Fl-Ox and Na-Cit do not stabilise Hcy in blood after collection and should not be used as preservatives.


Advanced REACH Tool (ART): overview of version 1.0 and research needs.
This paper provides an outline of the Advanced REACH Tool (ART) version 1.0 and a discussion of how it could be further developed. ART is a higher tier exposure assessment tool that combines mechanistically modelled inhalation exposure predictions with available exposure data using a Bayesian approach. ART assesses exposure for scenarios across different plants and sites. Estimates are provided for different percentiles of the exposure distribution and confidence intervals around the estimate. It also produces exposure estimates in the absence of data, but uncertainty of the estimates will decrease when results of exposure measurements are included. The tool has been calibrated using a broad range of exposure data and provides estimates for exposure to vapours, mists, and dusts. ART has a robust and stable conceptual basis but will be refined in the future and should therefore be considered an evolving system. High-priority areas for future research are identified in this paper and include the integration of partially analogous measurement series, inclusion of company and site-specific assessments, user decision strategies linked to ART predictions, evaluation of validity and reliability of ART, exploring the possibilities for incorporating the dermal route and integration of ART predictions with tools for modelling internal dose. ART is initially developed in the scope of REACH but is equally useful for exposure assessment in other areas.


Isolation and characterization of a cDNA from the rat brain that encodes hemoprotein heme oxygenase-3.
Two isozymes of heme oxygenase (HO), HO-1 or HSP32 and the constitutive form HO-2, have been characterized to date. We report the discovery of a third protein species and refer to it as HO-3. HO-3 is the product of a single transcript of approximately 2.4 kb and can encode a protein of approximately 33 kDa. The HO-3 transcript is found in the spleen, liver, thymus, prostate, heart, kidney, brain and testis and is the product of a single-copy gene. The predicted amino acid structure of HO-3 differs from both HO-1 (HSP32) and HO-2 but is closely related to HO-2 (approximately 90%). Escherichia coli expressed and purified HO-3 protein does not cross react with polyclonal antibodies to either rat HO-1 or HO-2, is a poor heme catalyst, and displays hemoprotein spectral characteristics. The predicted protein has two heme regulatory motifs that may be involved in heme binding. These motifs and the hemoprotein nature of HO-3 suggest a potential regulatory role for the protein in cellular processes which are heme-dependent.


A pilot study of occupational and environmental risk factors for Parkinson's disease.
Increasingly, the etiology of Parkinson's disease (PD) has been linked to exposures to environmental toxicants. This epidemiologic pilot study used a self-administered questionnaire among 34 PD cases and 22 other neurology clinic control patients. All subjects were at least 40 years old. Risk factors investigated included occupation, well-water use, pesticide use, metal exposures, medical history, smoking, alcohol consumption, and drug use. Twenty-six percent of the male PD cases reported having been employed in farming versus eleven percent for male controls (OR = 3.1, 95% C.I. = 0.3 to 35). Sixteen percent of male cases versus none of the controls reported employment as welders. No clear trends involving exposure to either occupational or home pesticides emerged. In assessing occupational exposures to metals, aluminum and copper exposures tended to be more common among male cases than male controls. Additionally, as reported in other studies, smoking showed an inverse relationship with PD. Although the findings reported here are provocative, these results are statistically imprecise and must be interpreted cautiously because of the small number of subjects included in the study.


The financial status of departments of family medicine at US medical schools.
This report examined the financial health of departments of family medicine in US allopathic medical schools. We conducted a survey of departments of family medicine at US medical schools, using academic year 1997-1998 as the index year. A total of 52 (46%) of medical schools that have a department of family medicine responded to the survey. The survey examined sources of revenue and categories of expenditures. Analysis assessed the overall financial status of departments at that period of time. Responding departments of family medicine received 32% of their funding from state or university sources and an additional 32% of funding from clinical services. Grants and hospital support comprised another 17% each. Departments in public institutions received higher levels of support from hospitals (22% of revenue versus 8% for private schools). The overall balance sheets for departments of family medicine showed that 56% of departments have financial reserves, while 19% had no reserves but no debt. Twenty-five percent of all departments were in debt, including 2% with debt exceeding $1 million. The majority of departments of family medicine remain fiscally healthy, but these departments are dependent on funds from state and medical school sources. A substantial proportion of departments are in debt. Lower levels of grant support and the difficulty in increasing clinical revenue may create future funding problems for primary care faculty as medical schools increase dependence on these sources of income.


Purification and microRNA profiling of exosomes derived from blood and culture media.
Stable miRNAs are present in all body fluids and some circulating miRNAs are protected from degradation by sequestration in small vesicles called exosomes. Exosomes can fuse with the plasma membrane resulting in the transfer of RNA and proteins to the target cell. Their biological functions include immune response, antigen presentation, and intracellular communication. Delivery of miRNAs that can regulate gene expression in the recipient cells via blood has opened novel avenues for target intervention. In addition to offering a strategy for delivery of drugs or RNA therapeutic agents, exosomal contents can serve as biomarkers that can aid in diagnosis, determining treatment options and prognosis. Here we will describe the procedure for quantitatively analyzing miRNAs and messenger RNAs (mRNA) from exosomes secreted in blood and cell culture media. Purified exosomes will be characterized using western blot analysis for exosomal markers and PCR for mRNAs of interest. Transmission electron microscopy (TEM) and immunogold labeling will be used to validate exosomal morphology and integrity. Total RNA will be purified from these exosomes to ensure that we can study both mRNA and miRNA from the same sample. After validating RNA integrity by Bioanalyzer, we will perform a medium throughput quantitative real time PCR (qPCR) to identify the exosomal miRNA using Taqman Low Density Array (TLDA) cards and gene expression studies for transcripts of interest. These protocols can be used to quantify changes in exosomal miRNAs in patients, rodent models and cell culture media before and after pharmacological intervention. Exosomal contents vary due to the source of origin and the physiological conditions of cells that secrete exosomes. These variations can provide insight on how cells and systems cope with stress or physiological perturbations. Our representative data show variations in miRNAs present in exosomes purified from mouse blood, human blood and human cell culture media. Here we will describe the procedure for quantitatively analyzing miRNAs and messenger RNAs (mRNA) from exosomes secreted in blood and cell culture media. Purified exosomes will be characterized using western blot analysis for exosomal markers and PCR for mRNAs of interest. Transmission electron microscopy (TEM) and immunogold labeling will be used to validate exosomal morphology and integrity. Total RNA will be purified from these exosomes to ensure that we can study both mRNA and miRNA from the same sample. After validating RNA integrity by Bioanalyzer, we will perform a medium throughput quantitative real time PCR (qPCR) to identify the exosomal miRNA using Taqman Low Density Array (TLDA) cards and gene expression studies for transcripts of interest. These protocols can be used to quantify changes in exosomal miRNAs in patients, rodent models and cell culture media before and after pharmacological intervention. Exosomal contents vary due to the source of origin and the physiological conditions of cells that secrete exosomes. These variations can provide insight on how cells and systems cope with stress or physiological perturbations. Our representative data show variations in miRNAs present in exosomes purified from mouse blood, human blood and human cell culture media.


Seawater Desalination by Modified Membrane Distillation: Effect of Hydrophilic Surface Modifying Macromolecules Addition into PVDF Hollow Fiber Membrane.
Hollow fiber membranes of polyvinylidene fluoride (PVDF) were prepared by incorporating varying concentrations of hydrophilic surface-modifying macromolecules (LSMM) and a constant amount of polyethylene glycol (PEG) additives. The membranes were fabricated by the dry-wet spinning technique. The prepared hollow fiber membranes were dip-coated by hydrophobic surface-modifying macromolecules (BSMM) as the final step fabrication. The additives combination is aimed to produce hollow fiber membranes with high flux permeation and high salt rejection in the matter of seawater desalination application. This study prepares hollow fiber membranes from the formulation of 18 wt. % of PVDF mixed with 5 wt. % of PEG and 3, 4, and 5 wt. % of LSMM. The membranes are then dip-coated with 1 wt. % of BSMM. The effect of LSMM loading on hydrophobicity, morphology, average pore size, surface porosity, and membrane performance is investigated. Coating modification on LSMM membranes showed an increase in contact angle up to 57% of pure, unmodified PVDF/PEG membranes, which made the fabricated membranes at least passable when hydrophobicity was considered as one main characteristic. Furthermore, The PVDF/PEG/4LSMM-BSMM membrane exhibits 161 °C of melting point as characterized by the DSC. This value indicates an improvement of thermal behavior shows so as the fabricated membranes are desirable for membrane distillation operation conditions range. Based on the results, it can be concluded that PVDF/PEG membranes with the use of LSMM and BSMM combination could enhance the permeate flux up to 81.32 kg·m-2·h-1 at the maximum, with stable salt rejection around 99.9%, and these are found to be potential for seawater desalination application.


B cell Sirt1 deacetylates histone and non-histone proteins for epigenetic modulation of AID expression and the antibody response.
Activation-induced cytidine deaminase (AID) mediates immunoglobulin class switch DNA recombination (CSR) and somatic hypermutation (SHM), critical processes for maturation of the antibody response. Epigenetic factors, such as histone deacetylases (HDACs), would underpin B cell differentiation stage-specific AID expression. Here, we showed that NAD+-dependent class III HDAC sirtuin 1 (Sirt1) is highly expressed in resting B cells and down-regulated by stimuli inducing AID. B cell Sirt1 down-regulation, deprivation of NAD+ cofactor, or genetic Sirt1 deletion reduced deacetylation of Aicda promoter histones, Dnmt1, and nuclear factor-κB (NF-κB) p65 and increased AID expression. This promoted class-switched and hypermutated T-dependent and T-independent antibody responses or led to generation of autoantibodies. Genetic Sirt1 overexpression, Sirt1 boost by NAD+, or allosteric Sirt1 enhancement by SRT1720 repressed AID expression and CSR/SHM. By deacetylating histone and nonhistone proteins (Dnmt1 and NF-κB p65), Sirt1 transduces metabolic cues into epigenetic changes to play an important B cell-intrinsic role in modulating antibody and autoantibody responses.


CD19-CAR engineered NK-92 cells are sufficient to overcome NK cell resistance in B-cell malignancies.
Many B-cell acute and chronic leukaemias tend to be resistant to killing by natural killer (NK) cells. The introduction of chimeric antigen receptors (CAR) into T cells or NK cells could potentially overcome this resistance. Here, we extend our previous observations on the resistance of malignant lymphoblasts to NK-92 cells, a continuously growing NK cell line, showing that anti-CD19-CAR (αCD19-CAR) engineered NK-92 cells can regain significant cytotoxicity against CD19 positive leukaemic cell lines and primary leukaemia cells that are resistant to cytolytic activity of parental NK-92 cells. The 'first generation' CAR was generated from a scFv (CD19) antibody fragment, coupled to a flexible hinge region, the CD3ζ chain and a Myc-tag and cloned into a retrovirus backbone. No difference in cytotoxic activity of NK-92 and transduced αCD19-CAR NK-92 cells towards CD19 negative targets was found. However, αCD19-CAR NK-92 cells specifically and efficiently lysed CD19 expressing B-precursor leukaemia cell lines as well as lymphoblasts from leukaemia patients. Since NK-92 cells can be easily expanded to clinical grade numbers under current Good Manufactoring Practice (cGMP) conditions and its safety has been documented in several phase I clinical studies, treatment with CAR modified NK-92 should be considered a treatment option for patients with lymphoid malignancies.


24-hour blood pressure variability and treatment effect of intravenous alteplase in acute ischaemic stroke.
To assess the association between 24 h blood pressure variability (BPV) on functional outcome and treatment effect of intravenous alteplase in acute ischaemic stroke. In all patients with acute ischaemic stroke of unknown onset randomised in the WAKE-UP (Efficacy and Safety of magnetic resonance imaging [MRI]-based Thrombolysis in Wake-Up Stroke) trial, blood pressure (BP) was measured before randomisation and after initiation of treatment at regular intervals up to 24 hours. Individual BPV was measured by coefficient of variation (CV) of all BP values. Primary outcome measure was favourable outcome defined by a modified Rankin Scale (mRS) score 0 or 1 at 90 days after stroke. BP measurements were available for 498 of 503 patients randomised (177 women [35.5%], mean age [SD] of 65.2 [11.5] years). Systolic BPV was not associated with the treatment effect of thrombolysis (test for interaction, p = 0.46). The adjusted odds ratio (aOR) for favourable outcome with alteplase, adjusted for age, stroke severity and baseline BP on admission, did not show an association across the quintiles of increasing systolic BPV with an aOR 1.89 (95% confidence interval [CI], 0.76-4.70) in the lowest quintile to aOR 1.05 (95% CI, 0.43-2.56) in the highest quintile. Higher mean systolic BP was associated with a smaller treatment effect of thrombolysis with a significant interaction (p = 0.033). The aOR for favourable outcome with alteplase decreased with quintiles of increasing mean systolic BP from aOR 3.16 (95% CI, 1.26-7.93) in the lowest quintile to aOR 0.84 (95% CI, 0.34-2.10) in in the highest quintile. There was a significant interaction between mean systolic BP and treatment effect of thrombolysis with higher mean systolic BP being associated with poorer outcome. BPV was not associated with outcome after thrombolysis.ClinicalTrials.gov identifier NCT01525290.


Effect of photodynamic treatment of human endothelial cells on cell volume and cell viability.
Photodynamic therapy (PDT) has yielded promising results in the treatment of malignant tumors. However, the mechanisms leading to tumor destruction during PDT are still not completely understood. In addition to effects on the microcirculation, damage to cellular structures has been observed following exposure of cells to PDT. A phenomenon preceding these events might possibly be cell swelling. We therefore studied the influence of treatment with Photofrin(R) (PF) and laser light on volume changes and cell viability of endothelial cells. Endothelial cells were obtained from human umbilical cord veins (HUVEC) by an adaption of the method of Maruyama. After subcultivation the cells were harvested and transferred as a cell suspension into a specially designed incubation chamber. Cells received either PF in concentrations of 1.5 or 3.0 mu g/ml and laser illumination 60 min post incubation (630 nm; 40 mW/cm(2), 4 Joule), PF alone, or laser treatment only. Following start of PF incubation and after phototreatment cell samples were taken for volume measurements using flow cytometry, and for studies of cellular morphology using scanning electron microscopy. Simultaneously, cell viability was monitored by the trypan blue exclusion test and the colorimetric MTT assay. Both control groups, HUVEC receiving PF or laser treatment alone, revealed constant cell volumes and cell viability during the entire course of the experiment. After PDT (60 min post-incubation) with 1.5 and 3.0 mu g PF/ml cell volume of HUVEC was increased at 15 min to 122%+/-6% and 140%+/-10% of baseline (100%), at 60 min to 152%+/-9% and 134%+/-18%, respectively (p<0.01). The number of viable cells was significantly reduced of samples treated with 1.5 and 3.0 mu g PF/ml at 15 min after PDT to 81%+/-3% and 76%+/-10% of baseline (100%), at 60 min after PDT to 32%+/-14% and 20%+/-15%, respectively (p<0.01). Scanning electron microscopy of cells exposed to PDT following 60 min incubation with Photofrin (3.0 mu g/ml) revealed significant cell damage. At the highest PF concentration HUVEC showed loss of microvilli and formation of blebs on the cellular surface. Our study demonstrates that PDT induces a significant increase in endothelial cell volume and a loss of cell viability. We suggest that swelling and damage of endothelial cells following PDT is a primary event finally contributing to cessation of blood flow and subsequent necrosis of tumors.


Pathologic stage of prostatic ductal adenocarcinoma at radical prostatectomy: effect of percentage of the ductal component and associated grade of acinar adenocarcinoma.
It is unknown whether ductal adenocarcinomas are more aggressive when matched for Gleason score (assigning the ductal component as Gleason pattern 4). Moreover, little is known whether a certain percentage of the ductal component is needed to account for its more aggressive behavior. Of 18,552 radical prostatectomies performed from 1995 to 2008, 93 cases with a ductal adenocarcinoma component were identified. Cases were classified based on their ductal/acinar ratio (<10%; ≥10% and <50%; ≥50%). There was no difference in the distribution of Gleason score 3+4=7 versus 4+3=7 between ductal and nonductal tumors, such that cases were combined as Gleason score 7. There was no age, race, and serum prostate-specific antigen difference between patients with and without ductal adenocarcinoma. Cases with ductal adenocarcinoma were less likely to be organ confined (36.6% vs 65.6%) and more likely to show seminal vesicle invasion (SVI) (19.3% vs 5.3%), P<0.0001. There was no difference in lymph node metastases or positive margins between cases with and without ductal features. An increasing percentage of the ductal component correlated with an increased risk of extraprostatic extension (P=0.04) and SVI (P<0.0001). To account for overall different Gleason scores between ductal and nonductal cases, and the effect of differing percentages of ductal features as well, the following analysis was carried out. For Gleason score 7 cases and ≥10% ductal differentiation, cases with ductal features were more likely to have nonfocal extraprostatic extension (64.0%) versus cases without ductal features (34.7%), P=0.002. In this group, there was no statistically significant difference in SVI or lymph node involvement between Gleason score 7 ductal and nonductal tumors. For Gleason score 7 cases with <10% ductal features, there was no difference in pathologic stage versus nonductal cases. There was no difference in pathologic stage between ductal and nonductal cases for Gleason score 8 to 10 cases, regardless of the percentage of the ductal component. This study shows that ductal adenocarcinoma admixed with Gleason pattern 3 is more aggressive than Gleason score 7 acinar cancer, as long as the ductal component is ≥10%. In cases with a very minor ductal component, these differences are lost. In addition, Gleason score 8 to 10 tumors with ductal features are not significantly more aggressive that acinar Gleason score 8 to 10 cancers in which the pure high-grade tumor, regardless of ductal features, determines the behavior.


[The effect of hydrocortisone on the impulse activity of cat cerebral cortex neurons].
The pulse activity of the neurons (50 in all) was studied in the brain somatic cortex by glass microelectrodes in acute experiments on 22 cats after an intravenous injection of hydrocortisone-hemisuccinate in doses of 0.03-8.0 mg/kg. 36 of 50 neurons changes their activity after the injection. Three types of reactions prevailed: an increase of the pulse frequency, its rarefaction, and a two-phase reaction in which the activation was followed by depression. As a rule the reaction occurred 10 to 60 sec after cortisone injection; this suggests a possibility of the direct action of the hormone on the brain cortex. A possible mechanism of variable neuronal reactions to hydrocortisone is discussed.


[The feasibility study of using the dual stylet method as a bedside measure in facilitating insertion of the spiral distal end nasal-enteral feeding tube in critically ill patients].
To evaluate the feasibility of using the dual stylet method as a bedside measure after unsuccessful of the spiral distal end nasal-enteral feeding tubes into the duodenum in critically ill patients. Spiral distal end nasal-enteral feeding tubes were introduced into the stomach of 50 critically ill patients but unable to pass through the pylorus from July 2005 to March 2007. Under electrocardiographic monitoring, the dual stylet method was used as a bedside measure to facilitate the passage. The duration, successful ratio, and complication of the procedure were recorded. This procedure took an average time of (24.5+/-4.9) minutes. The success rate of passing through the pylorus was 82.0% (41/50). The success rate of the latter 25 cases treated from July 2006 to March 2007 was significantly higher than that of the former 25 cases treated from July 2005 to July 2006 [96.0% (24/25) vs. 68.0% (17/25), P<0.05]. The average insertion distance of the 41 successful cases was (85.3+/-2.9)cm. Heart rate(HR) during the procedure was (116.7+/-18.5) beats per minute, that before insertion was (107.6+/-14.2) beats per minute (P<0.01), respiratory rate (RR) was (22.4+/-4.6)breaths per minute during the procedure and (21.3+/-3.9)breaths per minute (P<0.01) before the procedure and mean arterial pressure (MAP) (86.7+/-10.7) mm Hg during and (82.0+/-7.7)mm Hg (1 mm Hg=0.133 kPa, P<0.01) before the procedure. But there was no change in arterial oxygen saturation (SaO(2)). No severe complication was noted. The dual stylet method can be used effectively and safely in critically ill patients as a bedside measure after placement of the spiral distal end nasal-enteral feeding tubes.


A direct comparison of the electrophysiological effects of transcranial direct and alternating current stimulation in healthy subjects.
Despite the clinical effectiveness of transcranial direct current stimulation (tDCS) and transcranial alternating current stimulation (tACS), the comparability of these interventions in neurophysiological aspects have not been thoroughly investigated. Thus, we aimed to directly compare the electrophysiological effects of single-session tDCS and gamma-tACS in healthy subjects, matching the intervention protocol as closely as possible. This was a randomized, double-blinded, and active-controlled study. Sixty healthy college students were enrolled in the study. Both tACS with, at 40 Hz frequency, and tDCS have the same current intensity (2 mA, 30 min) within the same target area (right and left dorsolateral prefrontal cortex). Resting-state electroencephalography (EEG) was recorded before and after single-session stimulation. Significant differences in theta, alpha, low-beta, and gamma frequencies were found between tDCS, tACS, and the sham groups. Low-beta source activity of the middle temporal gyrus was decreased only after an intervention with tACS. The present study indicates that tDCS and tACS resulted in an increased range of frequency activity, including slow- and fast-wave bands. Specifically, tDCS modulates the frontal region, while tACS modulates neural oscillations at the fronto-central, parietal, and temporal areas. The tACS also decreased low-beta source activity in the middle temporal gyrus. Identifying the common and unique EEG patterns of tDCS and tACS may help shed light on their potential clinical benefits and distinctive neuropathology in various clinical symptoms.


Light-Switchable Membrane Permeability in Giant Unilamellar Vesicles.
In this work, giant unilamellar vesicles (GUVs) were synthesized by blending the natural phospholipid 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) with a photoswitchable amphiphile (1) that undergoes photoisomerization upon irradiation with UV-A (E to Z) and blue (Z to E) light. The mixed vesicles showed marked changes in behavior in response to UV light, including changes in morphology and the opening of pores. The fine control of membrane permeability with consequent cargo release could be attained by modulating either the UV irradiation intensity or the membrane composition. As a proof of concept, the photocontrolled release of sucrose from mixed GUVs is demonstrated using microscopy (phase contrast) and confocal studies. The permeability of the GUVs to sucrose could be increased to ~4 × 10-2 μm/s when the system was illuminated by UV light. With respect to previously reported systems (entirely composed of synthetic amphiphiles), our findings demonstrate the potential of photosensitive GUVs that are mainly composed of natural lipids to be used in medical and biomedical applications, such as targeted drug delivery and localized topical treatments.