Mandibular deviations in TMD and non-TMD groups related to eye dominance and head posture.
This study was designed to determine whether eye-dominance affects head posture (rotation) and in turn, whether head posture is associated with mandibular frenum midline deviation, in both TMJ and control subjects. Eye dominance was determined using three tests: Porta, Hole, Point tests. Natural head posture was evaluated using the Arthrodial protractor. Mandibular frenum deviation was recorded as left, right or no deviation. Fifty female subjects were included in the study, 25 TMJ patients attending the Gelb Craniomandibular Pain Center and 25 non-TMJ control subjects. The findings indicate that eye dominance and direction of head rotation are strongly associated in both TMJ and control subjects. Further, in TMJ subjects mandibular deviation occurred in greater frequency than in controls and tends to occur in the contra lateral direction of head rotation.


Azithromycin in Pseudomonas aeruginosa biofilms: bactericidal activity and selection of nfxB mutants.
Azithromycin (AZM) has shown promising results in the treatment of Pseudomonas aeruginosa chronic lung infections such as those occurring in cystic fibrosis (CF) patients. We evaluated the effect of hypermutation and alginate hyperproduction on the bactericidal activity and resistance development to AZM in P. aeruginosa biofilms. Strains PAO1, its microcA mutant (PAOMA), and their respective mutS-deficient hypermutable derivatives (PAOMS and PAOMSA) were used. Biofilms were incubated with several AZM concentrations for 1, 2, 4, or 7 days, and the numbers of viable cells were determined. During the first 2 days, AZM showed bactericidal activity for all the strains, but in extended AZM incubation for strain PAOMS and especially strain PAOMSA, a marked increased in the number of viable cells was observed, particularly at 4 microg/ml. Biofilms formed by the lineages recovered from the 7-day experiments showed enhanced AZM resistance. Furthermore, most of the independent lineages studied, including those obtained from biofilms treated with AZM concentrations as low as 0.5 microg/ml, showed MexCD-OprJ hyperexpression and mutations in nfxB. The role of nfxB mutation in AZM resistance was further confirmed through the characterization of nfxB and mexD knockout mutants. Results from this work show that, although AZM exhibits bactericidal activity against P. aeruginosa biofilms, resistant mutants are readily selected and that, furthermore, they frequently show cross-resistance to other unrelated antipseudomonal agents such as ciprofloxacin or cefepime but hypersusceptibility to others such as imipenem or tobramycin. Therefore, these results should help guide the selection of appropriate antipseudomonal therapies in CF patients under AZM maintenance treatment.


MAPK-mediated phosphorylation of GATA-1 promotes Bcl-XL expression and cell survival.
In the interleukin 3-dependent hematopoietic cell line Ba/F3, inhibition of mitogen-activated protein kinase, a member of the MAPK/c-Jun N-terminal kinase/stress-activated protein kinase kinase family that plays an important role in cell growth and death control, rapidly leads to severe apoptosis. However, most of the antiapoptotic substrates of MAPK remain to be identified. Here we report that, upon interleukin-3 stimulation of Ba/F3 cells, the transcription factor GATA-1 is strongly phosphorylated at residue serine 26 by a MAPK-dependent pathway. Phosphorylation of GATA-1 increases GATA-1-mediated transcription of the E4bp4 survival gene without significantly changing the DNA-binding affinity of GATA-1. Further characterization of GATA-1 phosphorylation site mutants revealed that the antiapoptotic function of GATA-1 is strongly dependent upon its phosphorylation at the Ser-26 position and is probably mediated through its up-regulation of Bcl-X(L) expression. Taken together, our data demonstrate that MAPK-dependent GATA-1 phosphorylation is important for its transactivation of the E4bp4 gene, Bcl-X(L) expression and cell survival. Therefore, GATA-1 may represent a novel MAPK substrate that plays an essential role in a cytokine-mediated antiapoptotic response.


A sensitive TLRH targeted imaging technique for ultrasonic molecular imaging.
The primary goals of ultrasound molecular imaging are the detection and imaging of ultrasound contrast agents (microbubbles), which are bound to specific vascular surface receptors. Imaging methods that can sensitively and selectively detect and distinguish bound microbubbles from freely circulating microbubbles (free microbubbles) and surrounding tissue are critically important for the practical application of ultrasound contrast molecular imaging. Microbubbles excited by low-frequency acoustic pulses emit wide-band echoes with a bandwidth extending beyond 20 MHz; we refer to this technique as transmission at a low frequency and reception at a high frequency (TLRH). Using this wideband, transient echo, we have developed and implemented a targeted imaging technique incorporating a multifrequency colinear array and the Siemens Antares imaging system. The multifrequency colinear array integrates a center 5.4-MHz array, used to receive echoes and produce radiation force, and 2 outer 1.5-MHz arrays used to transmit low-frequency incident pulses. The targeted imaging technique makes use of an acoustic radiation force subsequence to enhance accumulation and a TLRH imaging subsequence to detect bound microbubbles. The radiofrequency (RF) data obtained from the TLRH imaging subsequence are processed to separate echo signatures between tissue, free microbubbles, and bound microbubbles. By imaging biotin-coated microbubbles targeted to avidin-coated cellulose tubes, we demonstrate that the proposed method has a high contrast-to-tissue ratio (up to 34 dB) and a high sensitivity to bound microbubbles (with the ratio of echoes from bound microbubbles versus free microbubbles extending up to 23 dB). The effects of the imaging pulse acoustic pressure, the radiation force subsequence, and the use of various slow-time filters on the targeted imaging quality are studied. The TLRH targeted imaging method is demonstrated in this study to provide sensitive and selective detection of bound microbubbles for ultrasound molecularly targeted imaging.


Heterozygote advantage at MHC DRB may influence response to infectious disease epizootics.
The effect of MHC polymorphism on individual fitness variation in the wild remains equivocal; however, much evidence suggests that heterozygote advantage is a major determinant. To understand the contribution of MHC polymorphism to individual disease resistance or susceptibility in natural populations, we investigated two MHC class II B loci, DQB and DRB, in the New Zealand sea lion (NZSL, Phocarctos hookeri). The NZSL is a threatened species which is unusually susceptible to death by bacterial infection at an early age; it has suffered three bacterial induced epizootics resulting in high mortality levels of young pups since 1997. The MHC DQB and DRB haplotypes of dead NZSL pups with known cause of death (bacteria, enteritis or trauma) were sequenced and reconstructed, compared to pups that survived beyond 2 months of age, and distinct MHC DRB allele frequency and genotype differences were identified. Two findings were striking: (i) one DRB allele was present only in dead pups, and (ii) one heterozygous DRB genotype, common in live pups, was absent from dead pups. These results are consistent with some functional relationship with these variants and suggest heterozygote advantage is operating at DRB. We found no association between heterozygosity and fitness at 17 microsatellite loci, indicating that general heterozygosity is not responsible for the effect on fitness detected here. This result may be a consequence of recurrent selection by multiple pathogen assault over recent years and highlights the importance of heterozygote advantage at MHC as a potential mechanism for fitness differences in wild populations.


Rate and pattern of neuroretinal rim area decrease in ocular hypertension and glaucoma.
The neuroretinal rim areas of 123 eyes (from five normal subjects, 75 patients with ocular hypertension, and 43 patients with glaucoma) were measured to determine the rate and pattern of rim area change during 5 to 15 years (mean, 10 years) of follow-up. Fifty-seven percent of the patients with ocular hypertension and 79% of those with glaucoma showed a statistically significant slope of rim area decrease, a high rate of loss being associated with a high initial rim area. Ninety percent of the variation of rim area loss was accounted for by variables other than the ones measured herein, however (age, disc area, initial rim area, and intraocular pressure). The yearly loss of rim area was 0.23% of the initial area in normal subjects, 0.47% and 2.75% in the patients with stable and deteriorating ocular hypertension, respectively, and 3.47% in the patients with deteriorating glaucoma. The pattern of rim area change was linear in 49% of the patients with a statistically significant rim area decrease, episodic in 22%, and curvilinear in 29%.


Attachment and biofilm forming capabilities of Staphylococcus epidermidis strains isolated from preterm infants.
Staphylococcus epidermidis, a human commensal, is an important opportunistic, biofilm-forming pathogen and the main cause of late onset sepsis in preterm infants, worldwide. In this study we describe the characteristics of S. epidermidis strains causing late onset (>72 h) bloodstream infection in preterm infants and skin isolates from healthy newborns. Attachment and biofilm formation capability were analyzed in microtiter plates and with transmission electron microscopy (TEM). Clonal relationship among strains was studied with pulsed-field gel electrophoresis. Antimicrobial susceptibility testing was performed, as well as the detection of biofilm-associated genes and of the invasiveness marker IS256 with polymerase chain reaction. Blood and skin isolates had similar attachment and biofilm-forming capabilities and biofilm formation was not related to the presence of specific genes. Filament-like membrane structures were seen by TEM early in the attachment close to the device surface, both in blood and skin strains. Nine of the ten blood isolates contained the IS256 and were also resistant to methicillin and gentamicin in contrast to skin strains. S. epidermidis strains causing bloodstream infection in preterm infants exhibit higher antibiotic resistance and are provided with an invasive genetic equipment compared to skin commensal strains. Adhesion capability to a device surface seems to involve bacterial membrane filaments.


An Ensemble of Transfer Learning Models for the Prediction of Skin Cancers with Conditional Generative Adversarial Networks.
Skin cancer is one of the most severe forms of the disease, and it can spread to other parts of the body if not detected early. Therefore, diagnosing and treating skin cancer patients at an early stage is crucial. Since a manual skin cancer diagnosis is both time-consuming and expensive, an incorrect diagnosis is made due to the high similarity between the various skin cancers. Improved categorization of multiclass skin cancers requires the development of automated diagnostic systems. Herein, we propose a fully automatic method for classifying several skin cancers by fine-tuning the deep learning models VGG16, ResNet50, and ResNet101. Prior to model creation, the training dataset should undergo data augmentation using traditional image transformation techniques and Generative Adversarial Networks (GANs) to prevent class imbalance issues that may lead to model overfitting. In this study, we investigate the feasibility of creating dermoscopic images that have a realistic appearance using Conditional Generative Adversarial Network (CGAN) techniques. Thereafter, the traditional augmentation methods are used to augment our existing training set to improve the performance of pre-trained deep models on the skin cancer classification task. This improved performance is then compared to the models developed using the unbalanced dataset. In addition, we formed an ensemble of finely tuned transfer learning models, which we trained on balanced and unbalanced datasets. These models were used to make predictions about the data. With appropriate data augmentation, the proposed models attained an accuracy of 92% for VGG16, 92% for ResNet50, and 92.25% for ResNet101, respectively. The ensemble of these models increased the accuracy to 93.5%. A comprehensive discussion on the performance of the models concluded that using this method possibly leads to enhanced performance in skin cancer categorization compared to the efforts made in the past.


O-acetylation of a cell-surface carbohydrate creates discrete molecular patterns during neural development.
The cell-surface antigen detected by the monoclonal antibody JONES is expressed in the retina and a number of other central nervous system regions of the rat during the latter part of embryonic development and the early postnatal period. In addition to the expression on certain neuroblast populations it is found on some but not all axons and is also expressed at high levels on the end feet of radial glia in regions through which axons actively grow. In the perinatal rat retina, almost all the antigenic activity was carried on a ganglioside migrating between GM1 and GM2. The epitope recognized by antibody JONES was base labile and treatment with 0.1 M sodium carbonate or ammonia vapor converted the antigen into GD3. Resistance to oxidation by sodium periodate and reformation of the epitope by chemical acetylation of base-treated gangliosides with N-acetylimidazole identify the antigen as 9-O-acetyl GD3. The acetylation of GD3 seems to be regulated independently from GD3 expression itself since acetylated and nonacetylated GD3 do not have identical immunocytochemical distributions in the developing central nervous system. In addition, five independent human melanoma cell lines varied substantially in their expression of 9-O-acetyl GD3, even though they all expressed high levels of GD3. Acetylation of ganglioside-linked sialic acid provides a mechanism for generating unique patterns of surface carbohydrates, which may influence cell interactions in development.


Activin B is produced early in antral follicular development and suppresses thecal androgen production.
Little is known about the role of activin B during folliculogenesis. This study investigated the expression levels of activin/inhibin subunits (βA, βB, and α), steroid enzyme, and gonadotrophin receptors in theca (TC) and granulosa cells (GC) by QPCR and activin A and B and inhibin A protein levels in follicular fluid (FF) of developing sheep follicles during estrus and anestrus. The effect of activin B on androgen production from primary TC cultures in vitro was also assessed. During folliculogenesis, in anestrus and estrus, FF activin B concentrations and thecal and GC activin βB mRNA levels decreased as follicle diameter increased from 1-3 to >6 mm regardless of estrogenic status. Estrogenic preovulatory follicles had reduced concentrations of FF activins B and A, and TC and GCs expressed higher levels of activin βA mRNA at 3-4 mm, and TCs more inhibin α mRNA at >4 mm stages of development compared with nonestrogenic follicles. Activin B decreased androstenedione production from primary TCs in vitro, an effect blocked by inhibin A. Thus, sheep follicles 1-3 mm in diameter contained high FF levels of activin B, which decreased as the follicle size increased, and, like activin A, suppressed thecal androgen production in vitro, an effect blocked by inhibin. Furthermore, the theca of large estrogenic follicles expressed high levels of inhibin α and activin βA mRNA suggesting local thecal derived inhibin A production. This would inhibit the negative effects of thecal activins B and A ensuring maximum androgen production for enhanced estradiol production by the preovulatory follicle(s).


Inhibition of mammary carcinogenesis by systemic interleukin 12 or p185neu DNA vaccination in Her-2/neu transgenic BALB/c mice.
Because BALB/c mice transgenic for the rat Her-2/neu oncogene develop multifocal carcinomas in all mammary glands by week 33, they constitute an aggressive model for investigation of treatments designed to oppose mammary carcinogenesis. Nonspecific immune reaction elicited by systemic interleukin (IL)-12 both delayed the appearance of the first tumor and reduced the number of glands affected. However, only 5% of mice were tumor free at week 33. On the other hand, specific vaccination with plasmids encoding for the rat p185neu resulted in a further delay, so much so that 58% of mice were tumor free at week 33. No CTL response was evoked in either IL-12-treated or DNA-vaccinated mice, whereas an anti-rat p185neu antibody response was evident in the latter. Pathological examinations showed that in both IL-12-treated and DNA-vaccinated mice, the tumor growth area was infiltrated by reactive cells associated with expression of endothelial adhesion molecules and antiangiogenic proinflammatory cytokines. In the vaccinated mice, reduction of the number of cells expressing rat p185neu was combined with down-regulation of its membrane expression and even a marked inhibition in development of the terminal ductal lobular units. The reactive infiltrate in vaccinated mice contained numerous granulocytes that likely played an antiangiogenic and angiodestructive role and also joined other cells in the antibody-mediated killing of the r-p185neu+ cells. These results suggest that the elicitation of nonspecific and specific immunity could be beneficially used in individuals with a high risk of developing tumors.


Two distinct actin-binding sites of smooth muscle calponin.
Amino acid residues 145-163 of calponin have been proposed as a putative actin-binding site [Mezgueldi, M., Mendre, C., Calas, B., Kassab, R. & Fattoum, A. (1995) J. Biol. Chem. 270, 8867-8876]. Our previous work demonstrated that a fragment of calponin, which corresponded to the first repeated region of calponin and contained the preferred site of phosphorylation by protein kinase C [Nakamura, F., Mino, T., Yamamoto, J., Naka, M. & Tanaka, T. (1993) J. Biol. Chem. 268, 6194-6201] enhanced the Ca2+-induced contraction of permeabilized smooth muscle [Itoh, T., Suzuki, A., Watanabe, Y., Mino, T., Naka, M. & Tanaka, T. (1995) J. Biol. Chem. 270, 20400-20403]. In the present study, we compared the interactions with actin of a synthetic peptide (Lys172-His187) that encompassed the first repeated region with those of three other synthetic peptides. Lys172-His187 inhibited the binding of calponin to F-actin in a concentration-dependent manner but not the binding of caldesmon. Gly141-Gly160, including the above-mentioned putative actin-binding site, also competed with intact calponin to the same extent as Lys172-His187. Inhibition of actomyosin MgATPase activity was observed only with Gly141-Gly160. Lys172-His187 and other tested peptides had no effect. However, Gly141-Gly160 and Lys172-His187 reduced the fluorescence intensity of pyrene-labeled F-actin with approximately equal potency. Moreover, Lys172-His187 was able to reverse the inhibition of actomyosin MgATPase activity by calponin. Lys172-His187 was phosphorylated stoichiometrically by protein kinase C and phosphorylation of this peptide decreased its actin-binding activity. These observations suggest the direct involvement of two distinct actin-binding sites, with different regulatory functions, in the interactions of calponin with actin.


Evaluation of transmission route and replication efficiency of H9N2 avian influenza virus.
A/Chicken/Beijing/1/94 (Ck/BJ/1/94) avian influenza virus (AIV), a prototype of the H9N2 subtype, is phylogenetically similar in its hemagglutinin (HA) and neuraminidase (NA) genes to A/Chicken/Shanghai/F/98 (Ck/SH/F/98; H9N2) AIV, a natural reassortant between different sublineages. To understand the role of HA and NA genes in the airborne transmission of H9N2 AIV, we compared the transmission route and the relative replication efficiency of these strains in specific-pathogen-free chickens. Three recombinant viruses were generated by reverse genetics, containing the HA and NA genes (or both) from A/Chicken/Guangdong/SS/94 (Ck/GD/SS/94), in a background of internal genes derived from Ck/SH/F/98. Inoculated chickens were kept in either direct or indirect contact with uninoculated chickens, and viral shedding and titers were monitored. The results showed that Ck/GD/SS/94 lacks the ability to be transmitted through indirect contact, while Ck/SH/F/98 could be transmitted indirectly. Recombinant virus (RF/SSHA), containing the internal genes of Ck/SH/F/98 and the HA gene of Ck/GD/ SS/94, resulted in decreased viral titers in lung tissue as compared to the parental strain. Interestingly, substituting the NA gene, or both the NA and HA genes, of Ck/SH/F/98 with that of Ck/GD/SS/94 completely abolished the airborne transmission of the recombinant RF/SSNA and RF/SSHA/SSNA. In conclusion, Ck/SH/F/98 acquired the ability of airborne transmission and replicated with a higher efficiency in the respiratory tract of the chickens. Our data indicated that the NA gene of Ck/SH/F/98 can affect virus replication and, therefore, indirectly affect the transmission for the gene constellations of these viruses.


mzStudio: A Dynamic Digital Canvas for User-Driven Interrogation of Mass Spectrometry Data.
Although not yet truly 'comprehensive', modern mass spectrometry-based experiments can generate quantitative data for a meaningful fraction of the human proteome. Importantly for large-scale protein expression analysis, robust data pipelines are in place for identification of un-modified peptide sequences and aggregation of these data to protein-level quantification. However, interoperable software tools that enable scientists to computationally explore and document novel hypotheses for peptide sequence, modification status, or fragmentation behavior are not well-developed. Here, we introduce mzStudio, an open-source Python module built on our multiplierz project. This desktop application provides a highly-interactive graphical user interface (GUI) through which scientists can examine and annotate spectral features, re-search existing PSMs to test different modifications or new spectral matching algorithms, share results with colleagues, integrate other domain-specific software tools, and finally create publication-quality graphics. mzStudio leverages our common application programming interface (mzAPI) for access to native data files from multiple instrument platforms, including ion trap, quadrupole time-of-flight, Orbitrap, matrix-assisted laser desorption ionization, and triple quadrupole mass spectrometers and is compatible with several popular search engines including Mascot, Proteome Discoverer, X!Tandem, and Comet. The mzStudio toolkit enables researchers to create a digital provenance of data analytics and other evidence that support specific peptide sequence assignments.


Do male and female soccer players differ in helping? A study on prosocial behavior among young players.
Acting prosocially can be quite challenging in one of the most salient intergroup contexts in contemporary society: Soccer. When winning is the ultimate goal, balancing self-interest with helping a fellow player in distress can be a tough decision; yet it happens. To date, we know little about what motivates soccer players to offer such help in the heat of the game. We propose that sex and what is at stake will matter in such prosocial dilemma situations. A pilot study (N = 107) indicated that female players may be more likely to help than male players, but this difference was only observed when the players are close to scoring position rather than far away from the goal (midfield). The main study (N = 366) finds that young soccer players show elevated inclinations to help in low-stakes situations, for example when their team is winning or when the outcome of the game seems pretty much decided. Contrariwise, helping intentions decline in high-stakes situations, for example when one's own team is losing, when one is close to a scoring position in the offense (rather than at the midfield), or when the outcome of the game is still uncertain. Furthermore, female players show somewhat greater inclinations to help than their male counterparts. The current data point at some differences for male and female soccer players, albeit small in effect size. In contrast, we conclude that especially quick cost-benefit judgments regarding the stakes can play a major role in decisions to help or not to help another player on the soccer field.


Liver disease in infancy: histological features and relationship to alpha-antitrypsin phenotype.
Sixty-nine specimens of liver tissue from 53 infants with neonatal hepatitis or its sequelae were examined without knowledge of the alpha1-antitrypsin phenotype. Distinctive, diastase-resistant, PAS-positive, pure magenta-coloured, sharply defined globules, 2-20 microns in diameter were found in periportal and paraseptal hepatocytes in all liver biopsies from eight alpha1-antitrypsin deficient (PiZZ) infants biopsied after the age of 12 weeks. Such globules were not seen in biopsies from PiZZ individuals aged less than 12 weeks nor in individuals of normal alpha1-antitrypsin phenotype (PiMM). No other specific histological abnormality was found in PiZZ individuals, but in them giant-cell transformation was infrequent and liver damage was more severe, three of 14 cases developing cirrhosis in contrast to four of 27 PiMM subjects. The pathogenesis of liver disease in PiZZ individuals is discussed.


Rituximab Monotherapy for Compressive Optic Neuropathy With Giant Ocular Adnexal Mucosa-Associated Lymphoid Tissue lymphoma.
A rare case of compressive optic neuropathy due to giant mucosa-associated lymphoid tissue lymphoma in the orbit was presented. A 87-year-old woman was aware of a slow progressive left ocular proptosis for 10 years and presented after becoming aware of a sudden progression of the proptosis accompanying visual disturbance over the previous 2 months. Orbital imaging and a biopsy of the tumor revealed a mucosa-associated lymphoid tissue lymphoma occupying her left orbit compressing and stretching the left optic nerve. Considering her age and the additional adverse effects of external beam radiation therapy to her damaged optic nerve, rituximab monotherapy was utilized. The intervention resulted in almost complete regression without any serious adverse effect, with left eye best-corrected visual acuity improving from 12/200 to 16/20. Rituximab monotherapy can be one of the first-choice treatment options for mucosa-associated lymphoid tissue lymphoma, especially in cases with the critical damage in the optic nerve.


Inflammation and autoimmunity caused by a SHP1 mutation depend on IL-1, MyD88, and a microbial trigger.
A recessive phenotype called spin (spontaneous inflammation) was induced by N-ethyl-N-nitrosourea (ENU) mutagenesis in C57BL/6J mice. Homozygotes display chronic inflammatory lesions affecting the feet, salivary glands and lungs, and antichromatin antibodies. They are immunocompetent and show enhanced resistance to infection by Listeria monocytogenes. TLR-induced TNF and IL-1 production are normal in macrophages derived from spin mice. The autoinflammatory phenotype of spin mice is fully suppressed by compound homozygosity for Myd88(poc), Irak4(otiose), and Il1r1-null mutations, but not Ticam1(Lps2), Stat1(m1Btlr), or Tnf-null mutations. Both autoimmune and autoinflammatory phenotypes are suppressed when spin homozygotes are derived into a germ-free environment. The spin phenotype was ascribed to a viable hypomorphic allele of Ptpn6, which encodes the tyrosine phosphatase SHP1, mutated in mice with the classical motheaten alleles me and me-v. Inflammation and autoimmunity caused by SHP1 deficiency are thus conditional. The SHP1-deficient phenotype is driven by microbes, which activate TLR signaling pathways to elicit IL-1 production. IL-1 signaling via MyD88 elicits inflammatory disease.


Background severity of asthma in children discharged from the emergency department.
Attendance at an Emergency Department (ED) with an acute attack of asthma may be indicative of undertreatment of persistent disease. However, many presentations are in children with infrequent episodic asthma. The aim of this study was to characterize the pattern of asthma of children discharged from ED to determine whether there was potential to improve underlying disease control. This was a cohort study. Three hundred and ten parental caretakers of 1 to 15-year-old children, attended and discharged from an ED with asthma, completed an asthma control questionnaire, an asthma knowledge questionnaire and a caregiver's quality of life questionnaire. Background severity of asthma was classified and medication history was assessed. Also included were those with their first attack of asthma. One hundred and thirty-two (43%) children had infrequent episodic asthma, 105 (34%) frequent episodic, 40 (13%) persistent asthma and 33 (11%) first attack asthma. Thirty-nine per cent of children were not receiving preventer therapy and this seemed appropriate; 14% of children with frequent episodic and persistent asthma were not receiving appropriate preventer therapy; and a further 34% had frequent symptoms despite receiving preventer therapy. We observed deficiencies in use of preventer medications, use of written asthma management plans and lack of parental knowledge in some children with established asthma who presented to an ED. There was also a large number of children who did not have frequent background symptoms or who presented with their first episode.


Influence of low-level stimulus features, task dependent factors, and spatial biases on overt visual attention.
Visual attention is thought to be driven by the interplay between low-level visual features and task dependent information content of local image regions, as well as by spatial viewing biases. Though dependent on experimental paradigms and model assumptions, this idea has given rise to varying claims that either bottom-up or top-down mechanisms dominate visual attention. To contribute toward a resolution of this discussion, here we quantify the influence of these factors and their relative importance in a set of classification tasks. Our stimuli consist of individual image patches (bubbles). For each bubble we derive three measures: a measure of salience based on low-level stimulus features, a measure of salience based on the task dependent information content derived from our subjects' classification responses and a measure of salience based on spatial viewing biases. Furthermore, we measure the empirical salience of each bubble based on our subjects' measured eye gazes thus characterizing the overt visual attention each bubble receives. A multivariate linear model relates the three salience measures to overt visual attention. It reveals that all three salience measures contribute significantly. The effect of spatial viewing biases is highest and rather constant in different tasks. The contribution of task dependent information is a close runner-up. Specifically, in a standardized task of judging facial expressions it scores highly. The contribution of low-level features is, on average, somewhat lower. However, in a prototypical search task, without an available template, it makes a strong contribution on par with the two other measures. Finally, the contributions of the three factors are only slightly redundant, and the semi-partial correlation coefficients are only slightly lower than the coefficients for full correlations. These data provide evidence that all three measures make significant and independent contributions and that none can be neglected in a model of human overt visual attention.


Characterization of a developmental root response caused by external ammonium supply in Lotus japonicus.
Plants respond to changes of nutrient availability in the soil by modulating their root system developmental plan. This response is mediated by systemic changes of the nutritional status and/or by local perception of specific signals. The effect of nitrate on Arabidopsis (Arabidopsis thaliana) root development represents a paradigm of these responses, and nitrate transporters are involved both in local and systemic control. Ammonium (NH(4)(+)) represents an important nitrogen (N) source for plants, although toxicity symptoms are often associated with high NH(4)(+) concentration when this is present as the only N source. The reason for these effects is still controversial, and mechanisms associating ammonium supply and plant developmental programs are completely unknown. We determined in Lotus japonicus the range of ammonium concentration that significantly inhibits the elongation of primary and lateral roots without affecting the biomass of the shoot. The comparison of the growth phenotypes in different N conditions indicated the specificity of the ammonium effect, suggesting that this was not mediated by assimilatory negative feedback mechanisms. In the range of inhibitory NH(4)(+) conditions, only the LjAMT1;3 gene, among the members of the LjAMT1 family, showed a strong increased transcription that was reflected by an enlarged topology of expression. Remarkably, the short-root phenotype was phenocopied in transgenic lines by LjAMT1;3 overexpression independently of ammonium supply, and the same phenotype was not induced by another AMT1 member. These data describe a new plant mechanism to cope with environmental changes, giving preliminary information on putative actors involved in this specific ammonium-induced response.


Rapid detection of rifampicin- and isoniazid-resistant Mycobacterium tuberculosis by high-resolution melting analysis.
We have developed a high-resolution melting (HRM) assay to scan for mutations in the rpoB, inhA, ahpC, and katG genes and/or promoter regions for the detection of rifampin and isoniazid resistance in Mycobacterium tuberculosis. For assay development, 23 drug-resistant isolates of M. tuberculosis having 29 different mutations, together with 40 drug-susceptible isolates, were utilized. All 29 mutations were accurately detected by our assay. We further validated the assay with a series of 59 samples tested in a blind manner. All sequence alterations that were within the regions targeted by the HRM assay were correctly identified. Compared against results of DNA sequencing, the sensitivity and specificity of our HRM assay were 100%. For the blinded samples, the specificities and sensitivities were 89.3% and 100%, respectively, for detecting rifampin resistance and 98.1% and 83.3%, respectively, for detecting isoniazid resistance, as isolates with mutations in regions not encompassed by our assay were not detected. A C-to-T sequence alteration at position -15 of the ahpC regulatory region, which was previously reported to be associated with isoniazid resistance, may possibly be a polymorphism, as it was detected in an isoniazid-susceptible M. tuberculosis isolate. HRM is a rapid, accurate, simple, closed-tube, and low-cost method. It is thus an ideal assay to be used in countries with a high prevalence of drug-resistant M. tuberculosis and where cost-effectiveness is essential. As a mutation-scanning assay for detecting drug-resistant M. tuberculosis, it can potentially lead to better treatment outcomes resulting from earlier treatment with the appropriate antibiotics.


Layered SnSe nano-plates with excellent in-plane anisotropic properties of Raman spectrum and photo-response.
In-plane anisotropy in optical, electronic and thermal properties of two-dimensional (2D) materials has attracted significant interest because of the huge potential applications for developing novel devices. In this work, outstanding angle-dependent Raman property of layered SnSe nano-plates is obtained via polarized Raman system and it is confirmed that the Raman polarization directions of two Ag modes (130 cm-1 and 150 cm-1) are consistent with specific crystalline directions (zigzag direction or armchair direction) of SnSe flakes under parallel polarization configuration at home temperature and low temperature. Furthermore, the SnSe nano-plate devices show excellent angle-resolved photo-response at home temperature and low temperature (150 K) with a 90° cycle period and the polarized directions are also along zigzag direction and armchair direction, which is ascribed to the unique in-plane asymmetric crystal structure. These prominent in-plane anisotropic properties provide a precise and rapid method to determine the crystal orientation of SnSe nano-flakes and open up the new applications of 2D asymmetric structure materials.


Dendritic cells endocytose Bacillus anthracis spores: implications for anthrax pathogenesis.
Phagocytosis of inhaled Bacillus anthracis spores and subsequent trafficking to lymph nodes are decisive events in the progression of inhalational anthrax because they initiate germination and dissemination of spores. Found in high frequency throughout the respiratory track, dendritic cells (DCs) routinely take up foreign particles and migrate to lymph nodes. However, the participation of DCs in phagocytosis and dissemination of spores has not been investigated previously. We found that human DCs readily engulfed fully pathogenic Ames and attenuated B. anthracis spores predominately by coiling phagocytosis. Spores provoked a loss of tissue-retaining chemokine receptors (CCR2, CCR5) with a concurrent increase in lymph node homing receptors (CCR7, CD11c) on the membrane of DCs. After spore infection, immature DCs displayed a mature phenotype (CD83(bright), HLA-DR(bright), CD80(bright), CD86(bright), CD40(bright)) and enhanced costimulatory activity. Surprisingly, spores activated the MAPK cascade (ERK, p38) within 30 min and stimulated expression of several inflammatory response genes by 2 h. MAPK signaling was extinguished by 6 h infection, and there was a dramatic reduction of secreted TNF-alpha, IL-6, and IL-8 in the absence of DC death. This corresponded temporally with enzymatic cleavage of proximal MAPK signaling proteins (MEK-1, MEK-3, and MAP kinase kinase-4) and may indicate activity of anthrax lethal toxin. Taken together, these results suggest that B. anthracis may exploit DCs to facilitate infection.


Random walks based multi-image segmentation: Quasiconvexity results and GPU-based solutions.
We recast the Cosegmentation problem using Random Walker (RW) segmentation as the core segmentation algorithm, rather than the traditional MRF approach adopted in the literature so far. Our formulation is similar to previous approaches in the sense that it also permits Cosegmentation constraints (which impose consistency between the extracted objects from ≥ 2 images) using a nonparametric model. However, several previous nonparametric cosegmentation methods have the serious limitation that they require adding one auxiliary node (or variable) for every pair of pixels that are similar (which effectively limits such methods to describing only those objects that have high entropy appearance models). In contrast, our proposed model completely eliminates this restrictive dependence -the resulting improvements are quite significant. Our model further allows an optimization scheme exploiting quasiconvexity for model-based segmentation with no dependence on the scale of the segmented foreground. Finally, we show that the optimization can be expressed in terms of linear algebra operations on sparse matrices which are easily mapped to GPU architecture. We provide a highly specialized CUDA library for Cosegmentation exploiting this special structure, and report experimental results showing these advantages.


Clinical outcome of autologous mononuclear cells transplantation for spinal cord injury: a systematic review and meta-analysis.
Cell-based therapies can be used to treat neurological diseases and spinal cord injuries. The aim of this study was to assess the clinical outcome of bone marrow derived mononuclear cells (BM-MNCs) transplantation in patients with spinal cord injuries. Following a systematic review to detect clinical intervention studies, a meta-analysis was done for pooling data to estimate the outcome of BM-MNCs transplantation. The percentage of the patients with improved ASIA scale from one grade to a higher grade was defined as the main outcome. By considering the study design and outcome measurement, two reviewers independently extracted the data. Eight relevant primary studies were found; seven qualified studies, with a combined total of 328 patients were assessed by meta-analysis, including 314 ASIA-A, 13 ASIA-B, 94 cervical, 227 thoracic and 60 acute injuries. The percentage of the patients' improvement was tested by meta-analysis through random and fixed models. The overall percentage of all patients' improved ASIA scale after a one- year follow-up (95% CIs) was 43 (0.27-0.59). Data from published trials revealed that encouraging results were achieved by autologous BMMNCs for the treatment of spinal cord injury. However, the number of clinical trials included in the systematic review was too limited to reach a definite conclusion. More qualified clinical trials with standardized methods are needed to truly justify the outcome of this therapeutic modality in SCI patients.


A vesicular Warburg effect: Aerobic glycolysis occurs on axonal vesicles for local NAD+ recycling and transport.
In neurons, fast axonal transport (FAT) of vesicles occurs over long distances and requires constant and local energy supply for molecular motors in the form of adenosine triphosphate (ATP). FAT is independent of mitochondrial metabolism. Indeed, the glycolytic machinery is present on vesicles and locally produces ATP, as well as nicotinamide adenine dinucleotide bonded with hydrogen (NADH) and pyruvate, using glucose as a substrate. It remains unclear whether pyruvate is transferred to mitochondria from the vesicles as well as how NADH is recycled into NAD+ on vesicles for continuous glycolysis activity. The optimization of a glycolytic activity test for subcellular compartments allowed the evaluation of the kinetics of vesicular glycolysis in the brain. This revealed that glycolysis is more efficient on vesicles than in the cytosol. We also found that lactate dehydrogenase (LDH) enzymatic activity is required for effective vesicular ATP production. Indeed, inhibition of LDH or the forced degradation of pyruvate inhibited ATP production from axonal vesicles. We found LDHA rather than the B isoform to be enriched on axonal vesicles suggesting a preferential transformation of pyruvate to lactate and a concomitant recycling of NADH into NAD+ on vesicles. Finally, we found that LDHA inhibition dramatically reduces the FAT of both dense-core vesicles and synaptic vesicle precursors in a reconstituted cortico-striatal circuit on-a-chip. Together, this shows that aerobic glycolysis is required to supply energy for vesicular transport in neurons, similar to the Warburg effect.


Cancer-associated mutations in healthy individuals: assessing the risk of carcinogenesis.
Mutations associated with hematopoietic malignancies have been repeatedly identified in healthy individuals. For certain cases, such as the t(14;18) translocation and monoclonal B-cell lymphocytosis, no clear link between the presence of aberrant cells and the later development of cancer has been established. Intriguingly, longitudinal studies suggest that these abnormalities persist for long periods of time in some individuals, but in others are transient in which they disappear completely. Here, we present a mathematical model, based on cellular replication limits, that provides a possible explanation for these seemingly contradictory findings. It proposes that the transient and persistent nature of the phenotypes depends on the stage in the differentiation pathway of a given lineage in which the mutation originates. Our work suggests that cellular replication limits may not only prevent cancer by aborting clonal expansion of cells, but also by influencing the fate of altered but nonneoplastic cells in healthy tissue.


Prevalence and clinical relevance of diffusion-weighted imaging lesions: The Rotterdam study.
To examine the association of diffusion-weighted image (DWI) lesions with markers of cerebrovascular disease, neurodegeneration, and cognitive functioning and to further explore the evolution of these DWI lesions and their link to risk of dementia, stroke, or TIA in the Rotterdam Study. Two thousand one hundred seventy-six participants with baseline MRI scans (assessed between January 2009 and December 2013) and data on incident clinical outcomes (until January 2016) were included. DWIs were inspected for presence of acute or subacute lesions. Markers of cerebrovascular disease, brain tissue segmentation, and microstructural integrity were collected. Cognition was assessed with a detailed neuropsychological test. Evolution of DWI lesions was evaluated on follow-up scans. Thirty-three individuals (1.5%) had ≥1 DWI lesions. Persons with lacunes, white matter hyperintensities (WMHs), and reduced white matter microstructural integrity were more likely to have DWI lesions. Persons with DWI lesions performed worse on Stroop test 1. For 17 of 33 persons, follow-up scans were available to determine lesion evolution. During a mean follow-up of 4.7 years, 58.8% of DWI lesions appeared as WMHs, 17.6% developed cavitation, 5.9% changed into cortical cerebral microinfarcts, and 17.6% disappeared. People with DWI lesions at baseline were at increased risk of strokes (hazard ratio 3.72, 95% confidence interval 1.35-10.27). Asymptomatic DWI lesions in community-dwelling persons are associated with markers of cerebral small vessel disease, reduced microstructural integrity, and worse cognition. Presence of DWI lesions increases the risk of further strokes. Future investigations will have to show whether screening and treating persons with DWI lesions can effectively reduce the burden of stroke.


Family carers' experiences and perceived roles in interprofessional collaborative practice in primary care: A constructivist grounded theory study.
Chronic conditions can lead to physical, cognitive and social decline; thus, increasing an individual's dependence on family who assist with activities of daily living. Interprofessional collaborative practice (IPCP), involving two or more health professionals working with the patient and their family, is one model of care for the high-quality management of individuals with chronic conditions in primary care. Nevertheless, family carers have reported a disconnect between themselves and healthcare providers in previous research. This study aimed to explore the experiences and perspectives of family carers for individuals with chronic conditions, regarding their involvement in IPCP. Aspects of constructivist grounded theory methodology were used. Family carers of individuals with chronic conditions were invited to participate in a one-on-one, semistructured interview about their experiences with IPCP in the care of their loved one. Interview transcripts were analysed using Charmaz's four-step iterative process: (1) line-by-line coding, (2) focused coding, (3) categorisation of codes and (4) potential theme and subtheme development with memo writing to support each phase of analysis. The research team collaborated on reflexivity exercises, the conceptualisation of categories and the development of themes. Constructivist data analysis of interviews (average 40 min) with 10 family carers resulted in two themes. (1) Stepping in for my loved one represents the notion that carers take on external roles on behalf of their loved ones (subthemes: working with interprofessional teams, supporting independence and learning as I go). (2) Taking on the carer role, represents the internal factors that influence the external roles described in theme 1 (subthemes: feeling obligated to be involved and changing relationship dynamics). This study outlines the external actions and internal influences on family carer involvement in an interprofessional team. The required knowledge and support to care for their loved ones is currently learned in an ad hoc manner, and carers' resources should be better promoted by health professionals. Additionally, the relationship dynamics between a carer and their loved one change as the carer becomes more involved in IPCP and influences how and the extent health professionals involve family carers. Carers were the study population involved in this qualitative study. Patient advocates who have chronic conditions, and are informal family carers, were involved in the creation and design of this study, including a review of the research question, participant information sheet and the interview guide.


Highly specific and reliable in vitro diagnostic analysis of memory T and B lymphocytes in a Swiss cohort of COVID-19 patients.
The SARS-CoV-2 pandemic has claimed many lives and disrupted the quality of life of most individuals. Diagnostic tests not only serve to confirm past exposure but can provide information crucial for guiding healthcare options for patients. Current diagnostic tests for the presence of the SARS-CoV-2 virus or anti-spike protein antibodies do not address the question whether longer lasting cellular immunity is mounted in most individuals. Using an activation marker flow cytometric immunoassay (SARS-CoV-2 lymphocytes analysis), we showed that both CD4+/CD8+ T cell and B cell activation differ between naïve and infected individuals up to 11 months after infection. On the basis of the specificity of this diagnostic tool for detecting both SARS-CoV-2-experienced T and B cells, we propose that this assay could benefit immunocompromised individuals who are unable to mount sustained antibody responses, by determining cellular immunity as possible partial protection, and for studying immune correlates of protection - thereby increasing knowledge of COVID-19 in a wider range of patient groups.


Protocol for a multi-site pilot and feasibility randomised controlled trial: Surgery versus PhysiothErapist-leD exercise for traumatic tears of the rotator cuff (the SPeEDy study).
Clinically, a distinction is made between types of rotator cuff tear, traumatic and non-traumatic, and this sub-classification currently informs the treatment pathway. It is currently recommended that patients with traumatic rotator cuff tears are fast tracked for surgical opinion. However, there is uncertainty about the most clinically and cost-effective intervention for patients with traumatic rotator cuff tears and further research is required. SPeEDy will assess the feasibility of a fully powered, multi-centre randomised controlled trial (RCT) to test the hypothesis that, compared to surgical repair (and usual post-operative rehabilitation), a programme of physiotherapist-led exercise is not clinically inferior, but is more cost-effective for patients with traumatic rotator cuff tears. SPeEDy is a two-arm, multi-centre pilot and feasibility RCT with integrated Quintet Recruitment Intervention (QRI) and further qualitative investigation of patient experience. A total of 76 patients with traumatic rotator cuff tears will be recruited from approximately eight UK NHS hospitals and randomly allocated to either surgical repair and usual post-operative rehabilitation or a programme of physiotherapist-led exercise. The QRI is a mixed-methods approach that includes data collection and analysis of screening logs, audio recordings of recruitment consultations, interviews with patients and clinicians involved in recruitment, and review of study documentation as a basis for developing action plans to address identified difficulties whilst recruitment to the RCT is underway. A further sample of patient participants will be purposively sampled from both intervention groups and interviewed to explore reasons for initial participation, treatment acceptability, reasons for non-completion of treatment, where relevant, and any reasons for treatment crossover. Research to date suggests that there is uncertainty regarding the most clinically and cost-effective interventions for patients with traumatic rotator cuff tears. There is a clear need for a high-quality, fully powered, RCT to better inform clinical practice. Prior to this, we first need to undertake a pilot and feasibility RCT to address current uncertainties about recruitment, retention and number of and reasons for treatment crossover. ClinicalTrials.gov ( NCT04027205 ) - Registered on 19 July 2019. Available via.


Why do multiple traits determine mating success? Differential use in female choice and male competition in a water boatman.
Mating success is often determined by multiple traits, but why this occurs is largely unknown. Much attention has been paid to female preferences for multiple traits, but surprisingly few researchers have addressed the possibility that multiple traits are important because they serve different functions in female choice and male-male competition. Differential trait function could result from a conflict of interest between the sexes or from constraints forcing the sexes to pay attention to different traits. I show that traits determined at distinct life-history stages differ in their importance in female choice and male-male competition in a water boatman Sigara falleni. Juvenile conditions determined body and foreleg pala size and were the main determinants of mating success under female choice, whereas adult conditions determined body mass and influenced mating success when male competition was included. This differential use of condition-dependent traits under the two selection regimes appeared to arise partly from a conflict between the sexes, since the two selection forces (female choice and male competition) conflict for selection on pala size, and partly from constraints, as females appeared unable to assess adult condition.


Enhanced targeted DNA methylation of the CMV and endogenous promoters with dCas9-DNMT3A3L entails distinct subsequent histone modification changes in CHO cells.
With the emergence of new CRISPR/dCas9 tools that enable site specific modulation of DNA methylation and histone modifications, more detailed investigations of the contribution of epigenetic regulation to the precise phenotype of cells in culture, including recombinant production subclones, is now possible. These also allow a wide range of applications in metabolic engineering once the impact of such epigenetic modifications on the chromatin state is available. In this study, enhanced DNA methylation tools were targeted to a recombinant viral promoter (CMV), an endogenous promoter that is silenced in its native state in CHO cells, but had been reactivated previously (β-galactoside α-2,6-sialyltransferase 1) and an active endogenous promoter (α-1,6-fucosyltransferase), respectively. Comparative ChIP-analysis of histone modifications revealed a general loss of active promoter histone marks and the acquisition of distinct repressive heterochromatin marks after targeted methylation. On the other hand, targeted demethylation resulted in autologous acquisition of active promoter histone marks and loss of repressive heterochromatin marks. These data suggest that DNA methylation directs the removal or deposition of specific histone marks associated with either active, poised or silenced chromatin. Moreover, we show that de novo methylation of the CMV promoter results in reduced transgene expression in CHO cells. Although targeted DNA methylation is not efficient, the transgene is repressed, thus offering an explanation for seemingly conflicting reports about the source of CMV promoter instability in CHO cells. Importantly, modulation of epigenetic marks enables to nudge the cell into a specific gene expression pattern or phenotype, which is stabilized in the cell by autologous addition of further epigenetic marks. Such engineering strategies have the added advantage of being reversible and potentially tunable to not only turn on or off a targeted gene, but also to achieve the setting of a desirable expression level.


Association of provider-patient visit frequency and patient outcomes on hemodialysis.
In January of 2004, the Centers for Medicare & Medicaid Services tied provider reimbursement for outpatient hemodialysis services to the number of provider-patient visits per month. We aimed to determine whether greater visit frequency associated with lower mortality and hospitalization rates among incident hemodialysis patients in a large, nationally representative contemporary cohort. Using US Renal Data System data for 130,892 patients who initiated in-center hemodialysis between October 1, 2003 and September 30, 2006, we determined associations between the frequency of provider visits and mortality, first hospitalization, multiple hospitalizations, and cause-specific hospitalizations. Our primary analysis used Cox proportional hazards models, but we also performed time-varying Cox proportional hazards and instrumental variable analyses. In the primary analysis, we did not detect a significance difference in mortality among patients with four provider visits per month compared with those patients with fewer provider visits (adjusted HR=0.98; 95% CI=0.96-1.01), but the risk for first hospitalization was 4% lower among those patients with more frequent visits (adjusted HR=0.96; 95% CI=0.95-0.97). The time-varying Cox analysis produced similar results. The fully adjusted instrumental variable analysis showed a 0.07% higher risk for death that was not statistically significant (P=0.88) but a significant 2.3% lower risk for first hospitalization (P=0.001) for patients with four provider visits per month. In summary, greater frequency of provider visits to hemodialysis patients associates with a small but significant reduction in hospitalizations, but it does not consistently associate with lower risk for death.


Ionic Liquid Dispersive LiquidLiquid Microextraction Method for the Determination of Irinotecan, an Anticancer Drug, in Water and Urine Samples Using UV-Vis Spectrophotometry.
A new, simple, efficient, and environmentally friendly ionic liquid dispersive liquid–liquid microextractionmethod was developed for the determination of irinotecan, an anticancer drug, in water and urine samples using UV-Vis spectrophotometry. The ionic liquid 1-hexyl-3-methylimidazolium hexafluorophosphate was used as the extraction solvent, and ethanol was used as the disperser solvent. The main parameters affecting the extraction efficiency, including sample pH, volume of the ionic liquid, choice of the dispersive solvent and its volume, concentration of NaCl, and extraction and centrifugation times, were investigated and optimized. The effect of interfering species on the recovery of irinotecan was also examined. Under optimal conditions, the LOD (3σ) was 48.7 μg/L without any preconcentration. Because the urine sample was diluted 10-fold, the LOD for urine would be 487 μg/L. However, this could be improved 16-fold if preconcentration using a 40 mL aliquot of the sample is used. The proposed method was successfully applied to the determination of irinotecan in tap water, river water, and urine samples spiked with 10.20 mg/L for the water samples and 8.32 mg/L for the urine sample. The average recovery values of irinotecan determined were 99.1% for tap water, 109.4% for river water, and 96.1% for urine.


Concurrent validation of Hutt's Bender Gestalt screening method for schizophrenia, depression, and brain damage.
The effectiveness of Hutt's Bender Gestalt scoring system in screening for schizophrenia, depression, and brain damage was investigated in a sample of mixed, psychiatric inpatients. Fifteen patients represented each diagnostic category. After an examination of each patient's Bender Gestalt protocol according to Hutt's criteria, it was found that the true positive and true negative success rates of the screening procedures for depression and brain damage were significantly higher than corresponding base rates. The Lacks Bender Gestalt screening method for brain damage was no more successful than the Hutt method in identifying true positives and true negatives. With the brain-damaged patients excluded, Hutt's screening procedures for both depression and schizophrenia resulted in significantly high rates of successful identification of true positives and true negatives. Moreover, these rates did not significantly differ from those determined by patients' Minnesota Multiphasic Personality Inventory (MMPI) T scores on corresponding scales.


Utility of a Nurse-Led Pathway for Patients with Acute Venous Thromboembolism Discharged on Rivaroxaban: A Prospective Cohort Study.
The highest risk of adverse events for patients with acute venous thromboembolism (VTE) is during the early anticoagulation period. However, no established model exists for early clinical monitoring of patients treated with non-vitamin K antagonist oral anticoagulants (NOACs). The authors' aim was to evaluate the utility of a nurse-led pathway to minimize adverse events in acute VTE patients starting on rivaroxaban. The rivaroxaban VTE treatment pathway is a prospective cohort study of consecutive patients with objectively confirmed VTE between July 2015 and May 2017. Primary outcome was the proportion of patients identified at major risk of adverse events (bleeding or recurrent VTE). Secondary outcomes were rates of interventions, major or clinically relevant nonmajor bleeding (CRNMB), recurrent VTE, and all-cause mortality at 90 days. Among 304 participants, 5% (n = 15) were identified to be at major and 9% (n = 28) at possible risk for adverse events. Appropriate interventions to prevent harm were required in 40 patients. Rates of major bleeding, CRNMB, recurrence, and all-cause mortality were 0.3% (95% confidence interval [CI]: 0.1-1.8), 7.2% (95% CI: 4.8-10.7), 1.0 (95% CI: 0.3-2.9), and 1.6% (95% CI: 0.7-3.8), respectively. In conclusion, following discharge of acute VTE patients, a nurse-led pathway identified one in seven (14%) patients at major or possible risk of adverse events. Preemptive interventions to reduce harm translated into the low rates of bleeding and recurrence. The authors' experience highlights the feasibility and importance of a structured clinical surveillance pathway for acute VTE patients initiating NOAC therapy.


Diagnostic, demographic, memory quality, and cognitive variables associated with acute stress disorder in children and adolescents.
To date, no studies have investigated factors associated with acute stress disorder (ASD) in children and adolescents. Relationships between ASD and a number of demographic, trauma, cognitive, and trauma memory variables were therefore investigated in a sample (N=93) of children and adolescents involved in assaults and motor vehicle accidents. Several cognitive variables and the quality of trauma memories, but not demographic or trauma variables, were correlated with ASD and also mediated the relationship between peritraumatic threat and ASD. Finally, nosological analyses comparing ASD with indexes of posttraumatic stress disorder in the month posttrauma revealed little support for the dissociation mandate that uniquely characterizes ASD. The results are discussed with respect to assessment and treatment for the acute traumatic stress responses of children and young people.


The treatment of progressive pigmented purpura with ascorbic acid and a bioflavonoid rutoside.
Progressive pigmented purpura (PPP) is a group of dermatoses that are benign and usually self-limited. However, they may persist for months or years with frequent recurrences. Numerous treatments have been tried, but no effective therapy has yet proven to be successful. This report documents the treatment of a patient with one subtype of PPP using ascorbic acid and a bioflavonoid rutoside given orally. A 42-year-old male with biopsy-proven Eczematoid-like Purpura of Doucas and Kapetanakis was treated with ascorbic acid 500 mg twice a day and a bioflavonoid rutoside 50 mg twice a day. Treatment was successful, with quick defervescence of the rash and no recurrence to date.


Proteomic analysis of intestinal mucosa responses to Salmonella enterica serovar typhimurium in naturally infected pig.
Salmonella enterica serovar typhimurium (S. typhimurium) is one of the most frequent Salmonella serotypes isolated from European pigs. Despite the advances in understanding the mechanisms involved in host-pathogen interactions and host cell responses to S. typhimurium, the global change that occurs in naturally exposed populations has been poorly characterized. Here, we present a proteomics study on intestinal mucosa of pigs naturally infected with S. typhimurium, in order to better understand the pathogenesis of salmonellosis and the pathways which might be affected after infection. Samples were analyzed by 2D-DIGE and 44 different proteins exhibited statistically significant differences. The data set was analyzed by employing the Ingenuity Pathway Analysis and the physiological function most significantly perturbed were immunological and infectious disease, cellular assembly and organization and metabolism. The pathways implicated in the porcine immune response to S. typhimurium were gluconeogenesis and Rho GDI/RhoA signaling, and our results suggest that keratins and the intermediate filaments could play an important role in the damage of the mucosa and in the success of infection. The role of these findings in salmonellosis has been discussed, as well as the importance of analyzing naturally infected animals to have a complete picture of the infection. Also, we compared the results found in this work with those obtained in a similar study using experimentally infected animals.


Pulmonary rehabilitation: An overview.
The burden of chronic obstructive pulmonary disease (COPD) has increased recently in developing countries. On the other hand, structured or non-structured rehabilitation services for COPD patients are not routinely available in these countries. We, therefore, planned this review to re-emphasize the emerging benefits of pulmonary rehabilitation in COPD population. Aim of this review is to stimulate pulmonary physicians in India and other resource-poor areas of the world so that they start using pulmonary rehabilitation or its components more often. The search included standard english literature PubMed citation of relevant original articles, review articles and practice guidelines. The articles and reviews were searched including standard MeSH terms - Rehabilitation (TIAB) and pulmonary disease, chronic obstructive/therapy (MAJOR) and guidelines (TIAB). Available 58 articles in English including 23 reviews from July 2001 to October 2010 were screened for evidence-based benefits regarding respiratory rehabilitation as a whole or its different components. The cross references and current citations relating to primary articles were also included for description. No attempt was done to make a systematic analysis because our purpose was not to derive evidence-based recommendations from database and because sufficient evidence is already available for benefits of selected components of pulmonary rehabilitation in COPD patients. Pulmonary rehabilitation has emerged as an important modality as an adjunct to other therapies in patients of COPD. Limited and more cost-effective protocols are to be developed and executed by healthcare providers, especially in developing countries like India.


Error performances of a model-based biplane fluoroscopic system for tracking knee prosthesis during treadmill gait task.
Roentgen stereophotogrammetry analysis technique allows an accurate measurement of knee joint prosthesis position and orientation using two X-ray images. Although this technique is used generally during static procedure, it is possible to use it with a biplane fluoroscopic system to measure the prosthesis kinematics during functional tasks (e.g., gait, squat, jump) performed in a laboratory environment. However, the performance of the system in terms of errors for the measurements and the model-based matching algorithm are not well known for dynamic tasks such as walking. The goal of this study was to estimate the static and dynamic errors of a model-based biplane fluoroscopic system for a treadmill gait task and analyze the error performance according to the speed and location of the knee joint prosthesis relative to X-ray sources. The results show a static maximum error (RMSE) of 0.13° for orientation and 0.06 mm for position for prosthesis components. The dynamic errors were different for each axis of the acquisition system and each prosthesis component. The largest dynamic error was along the vertical axis for the position (RMSE = 2.42 mm) and along the medio-lateral axis (perpendicular to movement) for the orientation (RMSE = 0.95°). As expected, the error depends on the distance between the prosthesis and the source in the acquisition system as well as the linear and angular velocity of the movement. The most accurate dynamic measure was around the centroid of the acquisition system, while kinematics measurements close to the X-rays detectors gave the worst errors.


Characterization of the influenza virus M2 integral membrane protein and expression at the infected-cell surface from cloned cDNA.
An investigation of properties of the influenza A virus M2 protein indicated that it is synthesized by 2 h postinfection together with other viral polypeptides and is transported to the infected-cell surface with a half-time of approximately 30 to 40 min. The available evidence suggests that M2 is not N-glycosylated even though it contains a potential glycosylation site, and the intracellular pattern of protein distribution includes localization to the Golgi apparatus. Proteolysis of intracellular microsome vesicles followed by immunoprecipitation with antiserum to a synthetic oligopeptide indicated that the M2 protein contains an extensive region of COOH-terminal amino acids exposed on the cytoplasmic side of the infected-cell membrane. A cDNA clone of the M2 mRNA was obtained and expressed in an SV40 recombinant vector. The M2 protein expressed by the vector became associated with the Golgi complex and was found on the surface of vector-infected cells. M2 is antigenically conserved among all strains of influenza virus both in regions exposed on the cell surface and intracellularly.


Definition of a large region of RAG1 that is important for coimmunoprecipitation of RAG2.
Interaction between the RAG1 and RAG2 proteins is probably critical for V(D)J recombination. Using a coimmunoprecipitation assay, we define a large region of RAG1 (amino acids 504-1008) that is sufficient for interaction with RAG2. This region comprises the C-terminal half of the RAG1 protein, and is within the region defined as the recombinationally active core. Deletion of either of two regions of RAG1 (amino acids 504-570 or 850-1008) causes a loss of interaction with RAG2. Loss of coimmunoprecipitation is also seen with RAG1 core proteins containing deletions of smaller stretches of amino acids (amino acids 506-511 or 545-550), emphasizing the importance of this region of RAG1 in forming a complex with RAG2. A variety of other small deletion mutations within the amino acid region 504-1008 also decrease coimmunoprecipitation of RAG2 with RAG1, indicating that much or all of this region is important for complex formation.


[Evaluation of a rapid test (ImmunoCard STAT! HpSA) for Helicobacter pylori detection in stools].
The use of a rapid diagnostic technique may be extremely useful for the management of Helicobacter pylori infection. A new immunochromatographic in-office test (ImmunoCard STAT! HpSA, Meridian Diagnosis Inc, Cincinnati, Ohio, USA) for the detection of H. pylori in feces has recently become available. The aim of the present study was to evaluate the diagnostic reliability and reproducibility of the ImmunoCard STAT! HpSA test in patients with dyspepsia. Sixty-three dyspeptic patients were enrolled. H. pylori status was determined by CLO-test and Giemsa staining of antral biopsy. Patients with a positive result for both tests were considered infected and those with a negative result for both tests were considered not infected. Fecal H. pylori antigen was tested twice by ImmunoCard STAT! HpSA. The sensitivity, specificity, positive predictive value and negative predictive value of each determination were calculated. The concordance between the two determinations was evaluated using Kappa statistics. Of the 63 patients, 46 were infected by H. pylori. Sensitivity, specificity and positive and negative predictive values were 89-91%, 86-93%, 96-98% and 72-75%, respectively. The correlation coefficient between determinations was 0.845. The new ImmunoCard STAT! HpSA test shows good sensitivity and reproducibility. Therefore, it could be highly useful in the management of H. pylori infection.


Mouse models to study dengue virus immunology and pathogenesis.
The development of a compelling murine model of dengue virus (DENV) infection has been challenging, because DENV clinical isolates do not readily replicate or cause pathology in immunocompetent mice. However, research using immunocompromised mice and/or mouse-adapted viruses allows investigation of questions that may be impossible to address in human studies. In this review, we discuss the potential strengths and limitations of existing mouse models of dengue disease. Human studies are descriptive by nature; moreover, the strain, time, and sequence of infection are often unknown. In contrast, in mice, the conditions of infection are well defined and a large number of experimental parameters can be varied at will. Therefore, mouse models offer an opportunity to experimentally test hypotheses that are based on epidemiological observations. In particular, gain-of-function or loss-of-function models can be established to assess how different components of the immune system (either alone or in combination) contribute to protection or pathogenesis during secondary infections or after vaccination. In addition, mouse models have been used for pre-clinical testing of anti-viral drugs or for vaccine development studies. Conclusions based on mouse experiments must be extrapolated to DENV-infection in humans with caution due to the inherent limitations of animal models. However, research in mouse models is a useful complement to in vitro and epidemiological data, and may delineate new areas that deserve attention during future human studies.


The Adsorption Mechanism of Hydrogen on FeO Crystal Surfaces: A Density Functional Theory Study.
The hydrogen-based direct reduction of iron ores is a disruptive routine used to mitigate the large amount of CO2 emissions produced by the steel industry. The reduction of iron oxides by H2 involves a variety of physicochemical phenomena from macroscopic to atomistic scales. Particularly at the atomistic scale, the underlying mechanisms of the interaction of hydrogen and iron oxides is not yet fully understood. In this study, density functional theory (DFT) was employed to investigate the adsorption behavior of hydrogen atoms and H2 on different crystal FeO surfaces to gain a fundamental understanding of the associated interfacial adsorption mechanisms. It was found that H2 molecules tend to be physically adsorbed on the top site of Fe atoms, while Fe atoms on the FeO surface act as active sites to catalyze H2 dissociation. The dissociated H atoms were found to prefer to be chemically bonded with surface O atoms. These results provide a new insight into the catalytic effect of the studied FeO surfaces, by showing that both Fe (catalytic site) and O (binding site) atoms contribute to the interaction between H2 and FeO surfaces.


How the brain reacts to social stress (exclusion) - A scoping review.
The Cyberball paradigm is often used to study social stress by exclusion/rejection. We aimed to review the existing neuroimaging literatures in order to provide an overview of the neurophysiological mechanisms of social exclusion. Literature search was conducted to identify neurophysiological studies that investigated effects of social exclusion on neural activity using the Cyberball game and the relevant influential factors on these effects. In total, 42 studies using different neuroimaging methods were considered. Regions of the insula, anterior cingulate cortex, temporal and prefrontal cortex were activated to social exclusion. These neural activities were pronounced at latencies ranging from 200 to 400ms, and between 400 and 900ms. Influential factors were identified and categorized as intrinsic and extrinsic factors. An integrated framework describing neural activities under social exclusion in terms of both, temporal and spatial processes is provided. Furthermore, the summary of influential intrinsic and extrinsic factors may help us to understand the diversity of the processes and may guide clinical therapy of stress related disorders.


Implementation, Mechanisms and Context of the MAMAACT Intervention to Reduce Ethnic and Social Disparity in Stillbirth and Infant Health.
The MAMAACT intervention aimed to address ethnic and social disparity in stillbirth and infant health by improving management of pregnancy complications. This process evaluation of the intervention was guided by the British Medical Research Council's framework. We examined implementation through dose, reach, and fidelity, important mechanisms and the influence of contextual factors. The intervention included a six-hour training session for antenatal care (ANC) midwives in intercultural communication and cultural competence, two follow-up dialogue meetings, and health education materials (leaflet and app) on warning signs of severe pregnancy complications and how to respond for pregnant women. A mixed-methods approach was applied. Cross-sectional survey data and administrative data were used to assess intervention reach and dose. Qualitative data (records from dialogue meetings with midwives, participant observations and field notes from ANC visits, focus group interviews with midwives, and individual interviews with non-Western immigrant women) evaluated intervention fidelity, mechanisms, and contextual barriers. More than 80% of women received the MAMAACT leaflet and many found the content useful. The app was used more selectively. Midwives described being more aware and reflective in their communication with women from various cultural backgrounds. Organizational factors in ANC (time pressure, lack of flexibility in visits, poor interpreter services), barriers in women's everyday life (lack of social network, previous negative experiences/lack of trust and domestic responsibilities), and habitual interaction patterns among midwives served as contextual barriers. The reach of the intervention was high and it was evaluated positively by both pregnant women and midwives. Organizational factors hindered changes towards more needs-based communication in ANC potentially hindering the intended mechanisms of the intervention. When interpreting the intervention effects, attention should be drawn to both organizational and interpersonal factors in the clinic as well as the pregnant women's life situations.


Ornithine decarboxylase, the rate-limiting enzyme of polyamine synthesis, modifies brain pathology in a mouse model of tuberous sclerosis complex.
Tuberous sclerosis complex (TSC) is a rare autosomal dominant neurodevelopmental disorder characterized by variable expressivity. TSC results from inactivating variants within the TSC1 or TSC2 genes, leading to constitutive activation of mechanistic target of rapamycin complex 1 signaling. Using a mouse model of TSC (Tsc2-RG) in which the Tsc2 gene is deleted in radial glial precursors and their neuronal and glial descendants, we observed increased ornithine decarboxylase (ODC) enzymatic activity and concentration of its product, putrescine. To test if increased ODC activity and dysregulated polyamine metabolism contribute to the neurodevelopmental defects of Tsc2-RG mice, we used pharmacologic and genetic approaches to reduce ODC activity in Tsc2-RG mice, followed by histologic assessment of brain development. We observed that decreasing ODC activity and putrescine levels in Tsc2-RG mice worsened many of the neurodevelopmental phenotypes, including brain growth and neuronal migration defects, astrogliosis and oxidative stress. These data suggest a protective effect of increased ODC activity and elevated putrescine that modify the phenotype in this developmental Tsc2-RG model.


School-based interventions promoting both physical activity and healthy eating in Europe: a systematic review within the HOPE project.
It is the purpose of this study to systematically review the evidence of school-based interventions targeting dietary and physical activity behaviour in primary (6-12 years old) and secondary school (12-18 years old) children in Europe. Eleven studies (reported in 27 articles) met the inclusion criteria, six in primary school and five in secondary school children. Interventions were evaluated in terms of behavioural determinants, behaviour (diet and physical activity) and weight-related outcomes (body mass index [BMI] or other indicators of obesity). The results suggest that combining educational and environmental components that focus on both sides of the energy balance give better and more relevant effects. Furthermore, computer-tailored personalized education in the classroom showed better results than a generic classroom curriculum. Environmental interventions might include organized physical activities during breaks, or before and after school; improved availability of physical activity opportunities in and around the school environment; increased physical education lesson time; improved availability or accessibility of healthy food options; and restricted availability and accessibility of unhealthy food options. More high-quality studies are needed to assess obesity-related interventions in Europe.


Post-stroke fatigue: epidemiology, clinical characteristics and treatment.
Fatigue is a disabling and persistent symptom affecting many stroke survivors and is a predictor for death after stroke onset. Post-stroke fatigue is a multidimensional motor-perceptive, emotional and cognitive experience that has become of interest for stroke researchers only in the recent past. More studies are still needed to understand the pathophysiology, clinical characteristics, associated factors and best treatment strategy. The aim of this narrative review was to provide a comprehensive knowledge, from current literature, regarding the epidemiology, clinical characteristics and treatment of fatigue in order to provide physicians with better tools for treating this debilitating symptom that worsens outcome.


Skeletal muscle cytochrome c and myoglobin, endurance, and frequency of training.
This study was undertaken to evaluate the effects of various training frequencies on performance capacity, the mitochondrial marker cytochrome c, and myoglobin, which is responsible for storage and transport of O2, in the three types of skeletal muscle. Female rats were trained by treadmill running up to 120 min/day, either 2, 4, or 6 days/wk for 14 wk. As a result of training, exercise time to exhaustion was increased in proportion to the number of training sessions per week. Cytochrome c concentration increased (range 20-90%) as a linear function of the number of exercises per week in the fast-twitch red vastus lateralis (FTR), the slow-twitch soleus (STR), and the mixed plantaris muscles. However, the concentration of cytochrome c in fast-twitch white vastus lateralis (FTW) muscles increased to approximately the same extent (40-50%) in all training groups. The increases in myoglobin concentration (13-45%) with training were significantly related to frequency in FTR muscle but not in STR muscle. Myoglobin levels in FTW muscle remained unchanged, regardless of training group. These results provide evidence that the capacity to perform endurance exercise and the mitochondrial content of the red skeletal muscle types are directly affected by training frequency.


Takotsubo syndrome after liver transplantation: An association with intraoperatively administered epinephrine and fentanyl.
Takotsubo syndrome (TTS) can develop after liver transplant (LT), but its predisposing factors are poorly understood. In this study, we aimed to determine if perioperative factors were associated with posttransplant TTS. Adult patients who underwent primary LT between 2006 and 2018 were included. Patients with and without TTS were identified and matched by propensity scores. Of 2181 LT patients, 38 developed postoperative TTS with a mean left ventricular ejection fraction of 25.5% (±7.8%). Multivariable logistic regression revealed two preoperative risk factors (alcoholic cirrhosis and model for end-stage liver disease-sodium scores) for TTS. Post-propensity match analyses showed that TTS patients had significantly higher doses of epinephrine and lower doses of fentanyl during LT compared with non-TTS patients. A higher dose of epinephrine and a lower dose of fentanyl was associated with a higher predicted probability of TTS. All TTS patients had full recovery of cardiac function and had comparable 1-year survival. In conclusion, TTS occurred at a rate of 1.7% after LT and was associated with two pretransplant risk factors. The higher doses of epinephrine and lower doses of fentanyl administered during LT were associated with posttransplant TTS. More studies on the relationship between intraoperative medications and TTS are warranted.


Global CNS transduction of adult mice by intravenously delivered rAAVrh.8 and rAAVrh.10 and nonhuman primates by rAAVrh.10.
Some recombinant adeno-associated viruses (rAAVs) can cross the neonatal blood-brain barrier (BBB) and efficiently transduce cells of the central nervous system (CNS). However, in the adult CNS, transduction levels by systemically delivered rAAVs are significantly reduced, limiting their potential for CNS gene therapy. Here, we characterized 12 different rAAVEGFPs in the adult mouse CNS following intravenous delivery. We show that the capability of crossing the adult BBB and achieving widespread CNS transduction is a common character of AAV serotypes tested. Of note, rAAVrh.8 is the leading vector for robust global transduction of glial and neuronal cell types in regions of clinical importance such as cortex, caudate-putamen, hippocampus, corpus callosum, and substantia nigra. It also displays reduced peripheral tissue tropism compared to other leading vectors. Additionally, we evaluated rAAVrh.10 with and without microRNA (miRNA)-regulated expressional detargeting from peripheral tissues for systemic gene delivery to the CNS in marmosets. Our results indicate that rAAVrh.8, along with rh.10 and 9, hold the best promise for developing novel therapeutic strategies to treat neurological diseases in the adult patient population. Additionally, systemically delivered rAAVrh.10 can transduce the CNS efficiently, and its transgene expression can be limited in the periphery by endogenous miRNAs in adult marmosets.


Role of antibody and complement in the immune clearance and destruction of erythrocytes. II. Molecular nature of IgG and IgM complement-fixing sites and effects of their interaction with serum.
A model for the immune clearance and destruction of homologous erythrocytes has been further explored. In this model, every IgM anti-erythrocyte antibody molecule in an antibody preparation was shown to fix Cl. About 2000 IgG antibody molecules were required to form a Cl-fixing site on the guinea pig erythrocyte surface. 60 IgM complement-fixing sites per erythrocyte were required for the immune clearance of IgM-sensitized erythrocytes. This number of sites could be detected by a direct agglutination test. 1.4 complement-fixing sites were required for immune clearance of IgG-sensitized cells, a number of molecules which could not be detected by direct agglutination. This number could, however, be detected with the use of a Coombs antiglobulin reagent. Depletion of the late components of complement (C3-9) with cobra venom was associated with the loss of ability to clear IgM-sensitized cells and a marked deficit in the ability to clear IgG-coated cells. Thus, late (C3-9) components of complement as well as an early component (C4) were required for normal clearance of sensitized erythrocytes. There was no evidence that activation of the alternate pathway of complement action could lead to accelerated erythrocyte clearance. In vitro incubation of IgG and IgM-sensitized erythrocytes in fresh serum led to deposition of C3 and C4 on the erythrocyte surface. IgM-sensitized cells treated in this way had a normal survival. IgM-sensitized cells also were shown to remain Coombs positive after their release from the liver. The evidence suggests that the interaction of an IgM site with fresh serum in vitro and in vivo leads to formation of a site which allows for sequestration of cells in the liver. With continued exposure to serum components, this site is destroyed or inactivated. This serum-dependent inactivation is complement-dependent as shown by the use of EDTA-treated and C4-deficient serum. IgG complement-fixing sites are only partially inactivated by incubation in fresh serum, further emphasizing the differences in the biologic activity of IgM and IgG antibodies.


Deficiency of melanoma differentiation-associated protein 5 results in exacerbated chronic postviral lung inflammation.
Respiratory viral infections can result in the establishment of chronic lung diseases. Understanding the early innate immune mechanisms that participate in the development of chronic postviral lung disease may reveal new targets for therapeutic intervention. The intracellular viral sensor protein melanoma differentiation-associated protein 5 (MDA5) sustains the acute immune response to Sendai virus, a mouse pathogen that causes chronic lung inflammation, but its role in the development of postviral chronic lung disease is unknown. To establish the role of MDA5 in the development of chronic lung disease. MDA5-deficient or control mice were infected with Sendai virus. The acute inflammatory response was evaluated by profiling chemokine and cytokine expression and by characterizing the composition of the cellular infiltrate. The impact of MDA5 on chronic lung pathology and function was evaluated through histological studies, degree of oxygen saturation, and responsiveness to carbachol. MDA5 deficiency resulted in normal virus replication and in a distinct profile of chemokines and cytokines that associated with acute lung neutropenia and enhanced accumulation of alternatively activated macrophages. Diminished expression of neutrophil-recruiting chemokines was also observed in cells infected with influenza virus, suggesting a key role of MDA5 in driving the early accumulation of neutrophils at the infection site. The biased acute inflammatory response of MDA5-deficient mice led to an enhanced chronic lung inflammation, epithelial cell hyperplasia, airway hyperreactivity, and diminished blood oxygen saturation. MDA5 modulates the development of chronic lung inflammation by regulating the early inflammatory response in the lung.


Energy metabolism in shock.
Shock produces a state of low-energy production secondary to lack of oxygen. Only 1/5 of the energy produced under aerobic conditions can be produced in shock and less so if shock continues and loss of nucleotide bases and substrate occurs. Carbohydrates and their intermediate compounds are responsible for the energy produced during shock. As cellular derangements and loss of energy supplying compounds are becoming better understood, their contributions to a condition of irreversible shock are receiving more consideration. Studies utilizing energy substrates, prevention of nucleotide loss, and supplying high-energy compounds have been done and show promise as a therapeutic modality.


PDGF-dependent regulation of regulator of G protein signaling-5 expression and vascular smooth muscle cell functionality.
Regulator of G protein signaling (RGS) proteins, and notably members of the RGS-R4 subfamily, control vasocontractility by accelerating the inactivation of Gα-dependent signaling. RGS5 is the most highly and differently expressed RGS-R4 subfamily member in arterial smooth muscle. Expression of RGS5 first appears in pericytes during development of the afferent vascular tree, suggesting that RGS5 is a good candidate for a regulator of arterial contractility and, perhaps, for determining the mass of the smooth muscle coats required to regulate blood flow in the branches of the arterial tree. Consistent with this hypothesis, using cultured vascular smooth muscle cells (VSMCs), we demonstrate RGS5 overexpression inhibits G protein-coupled receptor (GPCR)-mediated hypertrophic responses. The next objective was to determine which physiological agonists directly control RGS5 expression in VSMCs. GPCR agonists failed to directly regulate RGS5 mRNA expression; however, platelet-derived growth factor (PDGF) acutely represses expression. Downregulation of RGS5 results in the induction of migration and the activation of the GPCR-mediated signaling pathways. This stimulation leads to the activation of mitogen-activated protein kinases directly downstream of receptor stimulation, and ultimately VSMC hypertrophy. These results demonstrate that RGS5 expression is a critical mediator of both VSMC contraction and potentially, arterial remodeling.


Endoplasmic reticulum stress in nonalcoholic (metabolic associated) fatty liver disease (NAFLD/MAFLD).
Nonalcoholic fatty liver disease (NAFLD) is characterized by hepatic fat accumulation in the absence of excessive alcohol consumption and is strongly associated with obesity, type 2 diabetes (T2DM) and other metabolic syndrome features. NAFLD is becoming increasingly prevalent and currently constitutes the leading cause of hepatocellular carcinoma (HCC). Recently, the term metabolic (dysfunction) associated fatty liver disease (MAFLD) has been proposed reflecting more accurately the underlying pathogenesis and the cardiometabolic disorders associated to NAFLD/MAFLD. Given the vital metabolic functions of the liver to maintain the body homeostasis, an extended endoplasmic reticulum (ER) network is mandatory in hepatocytes to retain its capacity to adapt to the multiple extracellular and intracellular signals mediating metabolic changes. Dysfunction of hepatocyte ER homeostasis and disturbance of its interaction with mitochondria have been recognized to be involved in the NAFLD pathophysiology. Apart from hepatocytes, hepatic stellate cells, and Kupffer cells have been shown to play an important role in the occurrence of NAFLD and progression to nonalcoholic steatohepatitis (NASH) with possibly different roles in the different stages of the NAFLD spectrum. Furthermore, excess lipid accumulation in the liver causes lipotoxicity which interacts with ER stress and culminates in inflammation and hepatocellular damage, mechanisms crucially implicated in NASH pathogenesis. Finally, the circadian clock machinery regulates ER stress-related pathways and vice versa, thus controlling the homeostasis of the liver metabolism and being implicated in the NAFLD progression. This review presents a comprehensive overview of the current knowledge supporting the impact of ER stress signaling on NAFLD, whilst summarizing potential therapeutic interventions targeting this process.


Sources of meaning in the acquisition of complex syntax: the sample case of causality.
The study reported here is concerned with how children acquire complex sentences for expressing their beliefs about causally related events, in the transition in language development from simple to complex syntax. Subjects were three girls and four boys, observed longitudinally from 26 to 38 months of age in their homes. Data analysis began with those observations in which each child began to produce causally related propositions without syntactic connectives, and continued until the children were about 3 years old. Two broad categories of causal meaning were expressed in the children's causal statements. Objective meaning concerned means-end and consequence relations that were evidential and fixed in the physical world. Subjective meaning expressed causal connections concerned with personal, affective, or sociocultural beliefs. While most of the children's statements expressed subjective meaning overall, the acquisition of syntactic connectives was associated with objective meaning. These results are discussed in terms of the development of these children's understanding of causality and the acquisition of increasingly complex language.


Evaluating a Novel Hemodialysis Central Venous Catheter Cap in Reducing Bloodstream Infections: A Quality Improvement Initiative.
Central line-associated bloodstream infection (CLABSI) is the second leading cause of death in hemodialysis patients. Patients dialyzed via central venous catheters (CVCs) are more susceptible to vascular access-related bloodstream infection (VRBSI), sepsis, and mortality when compared to patients with other dialysis accesses. A retrospective observational data analysis was conducted from 13 outpatient dialysis clinics in the United States to compare novel chlorhexidine-coated end caps to standard needlefree connectors for differences in CLABSI rates when utilizing CVCs for hemodialysis. There were two periods in this study: in the first study period over a 5-month period (May 2018 to September 2018), data were evaluated from a group of patients undergoing hemodialysis using chlorhexidine end-caps ('chlorhexidine group') as well as a group using standard needlefree connectors ('standard group'). An initial assessment found that a substantial CLABSI rate reduction was seen with use of chlorhexidine-coated end caps; therefore, most patients were switched to chlorhexidine by February 2019 and data continued to be collected till June 2019. The second study period spanned 9 months from October 2018 to June 2019. Across 13 dialysis centers, anonymized health records of 5934 patients who were dialyzed via CVCs between May 2018 and June 2019 were analyzed. The mean age was 61.3 and 47.1% of all patients were female. Study period one included 967 patients with chlorhexidine and 1044 patients with standard end caps, while there were 3647 chlorhexidine and 276 standard patients in the second period. The combined CLABSI rate in the chlorhexidine group was 0.09/1000 CVC days versus 0.63/1000 CVC days in the standard group (p<0.0001). Chlorhexidine-coated CVC caps may provide a therapeutic improvement in CVC hemodialysis management.