Stages of change in adults who have failed an online hearing screening.
Hearing screening has been proposed to promote help-seeking and rehabilitation in adults with hearing impairment. However, some longitudinal studies point to low help-seeking and subsequent rehabilitation after a failed hearing screening (positive screening result). Some barriers to help-seeking and rehabilitation could be intrinsic to the profiles and needs of people who have failed a hearing screening. Theories of health behavior change could help to understand this population. One of these theories is the transtheoretical (stages-of-change) model of health behavior change, which describes profiles and needs of people facing behavior changes such as seeking help and taking up rehabilitation. According to this model, people go through distinct stages toward health behavior change: precontemplation, contemplation, action, and finally, maintenance. The present study describes the psychometric properties (construct validity) of the stages of change in adults who have failed an online hearing screening. Stages of change were measured with the University of Rhode Island Change Assessment (URICA). Principal component analysis is presented, along with cluster analysis. Internal consistency was investigated. Finally, relationships between URICA scores and speech-in-noise recognition threshold, self-reported hearing disability, and self-reported duration of hearing disability are presented. In total, 224 adults who had failed a Swedish online hearing screening test (measure of speech-in-noise recognition) completed further questionnaires online, including the URICA. A principal component analysis identified the stages of precontemplation, contemplation, and action, plus an additional stage, termed preparation (between contemplation and action). According to the URICA, half (50%) of the participants were in the preparation stage of change. The contemplation stage was represented by 38% of participants, while 9% were in the precontemplation stage. Finally, the action stage was represented by approximately 3% of the participants. Cluster analysis identified four stages-of-change clusters: they were named decision making (44% of sample), participation (28% of sample), indecision (16% of sample), and reluctance (12% of sample). The construct validity of the model was good. Participants who reported a more advanced stage of change had significantly greater self-reported hearing disability. However, participants who reported a more advanced stage of change did not have a significantly worse speech-in-noise recognition threshold or reported a significantly longer duration of hearing impairment. The additional stage this study uncovered, and which other studies have also uncovered, preparation, highlights the need for adequate guidance for adults who are yet to seek help for their hearing. The fact that very few people were in the action stage (approximately 3% of the sample) signals that screening alone is unlikely to be enough to improve help-seeking and rehabilitation rates. As expected, people in the later stages of change reported significantly greater hearing disability. The lack of significant relationships between stages-of-change measures and speech-in-noise recognition threshold and self-reported duration of hearing disability highlights the complex interplay between impairment, disability, and behaviors in adults who have failed an online hearing screening and who are yet to seek help.


Virtual reality demonstration of surgical specimens, including links to histologic features.
The demonstration of surgical specimens, whether using 35-mm slides or digital images, tends to consist of the sequential presentation of images. Current digital technology permits a more flexible and effective way of communication, with the opportunity to more easily "navigate" between different aspects of specimens. We demonstrate a "virtual reality" method, based on QuickTime VR technology, that permits the interactive review of a complete profile of surgical specimens in the horizontal plane. Specimens were placed individually on a circular rotating platform. Thirty-six images of each specimen were captured using a digital camera, with rotation of the platform at 10 degrees intervals. The images were transferred to a computer and processed using specialized software (VRWorx). Histologic images were separately captured from tissue sections on glass slides using a digital camera mounted on a microscope. The final product is viewed using the QuickTime Viewer software application. A 360 degrees horizontal view of the specimens is achieved, with the capacity to actively rotate the specimen and to zoom in for closer review. Additionally, the user/presenter can click in predetermined "hot spots," which will open histologic images linked to those spots. This methodology, which uses readily available computer technology, helps provide a better three-dimensional understanding of surgical specimens and also a better correlation between gross and microscopic features.


Effects of partial hepatectomy on the distal ileum in rats.
hepatectomy, both partial resection and transplant, has greatly developed and advanced during the last years as a result of a better understanding of surgical anatomy and the progress of technological means; it has also allowed a widening of surgical indications, including living-donor liver transplantation. The aims of our study was to assess the morphological impact of partial hepatectomy on the distal ileum, since the liver and intestine behave as a unit from an anatomical, functional, and metabolic point of view. twenty-four Wistar rats were used; they were divided into two groups, a control and an experimental group (30, 90, and 180 days). We studied changes occurred in the distal ileum after a 70% liver resection, taking 4 parameters into account. an important drop in total thickness occurred at the ileum wall in the experimental group (p < 0.001). There were no important differences in villus height with regard to the control group. Thickness at the villi fell significantly (p < 0.01), as did the depth of the crypts, which diminished significantly in relation to the control group (p < 0.001). a 70% hepatectomy induces trophic changes on the distal ileum that remain in both the short and longer term, and causes atrophy of the ileum wall and a drop in villus thickness.


The effect of non-antihypertensive doses of angiotensin converting enzyme inhibitor on myocardial necrosis and hypertrophy in young rats with renovascular hypertension.
In renovascular hypertensive rats, low doses of angiotensin converting enzyme (ACE) inhibitors have been found to prevent myocardial hypertrophy independent of blood pressure level. This finding would suggest humoral rather than mechanical control of myocyte growth. The aim of this study was to examine the effect of nonantihypertensive doses of ACE inhibitor on myocardial hypertrophy and necrosis in hypertensive rats. Renovascular hypertension (RHT) was induced in four-week-old Wistar rats. Twenty-eight animals were treated for four weeks with three doses of ramipril (0.01, 0.1 or 1. 0 mg/kg/day, which are unable to lower blood pressure. Fourteen animals were not treated (RHT group). A sham operated, age/sex-matched group was used as control (n = 10). Myocardial histology was analysed in 3 microm thick sections of the ventricle stained with either haematoxylin-eosin, reticulin silver stain or Masson's trichrome. There was a significant correlation between systolic blood pressure and left ventricular to body weight ratio in both sets of animals: untreated plus controls and ramipril-treated rats. ACE inhibition prevented myocyte and perivascular necrosis and fibrosis in a dose-dependent manner. We conclude that myocardial hypertrophy in rats with renovascular hypertension is directly related to arterial pressure, and that this relationship is not affected by nonantihypertensive doses of ACE inhibitor. Myocardial necrosis/fibrosis and coronary artery damage induced by angiotensin II are prevented by ACE inhibitor in a dose-dependent manner, despite the presence of arterial hypertension.


Liquid-phase electron microscopy imaging of cellular and biomolecular systems.
The ongoing development of liquid-phase electron microscopy methods-in which specimens are kept fully solvated in the microscope by encapsulation in transparent, vacuum-tight chambers-is making it possible to investigate a wide variety of nanoscopic dynamic phenomena at the single-particle level, and with nanometer to atomic resolution. As such, there has been growing motivation to make liquid-phase electron microscopy tools applicable not only to inorganic materials, like metals, semiconductors, and ceramics, but also to "soft" materials such as biomolecules and cells, whose nanoscale dynamics and organization are intricately tied to their functionality. Here we review efforts toward making this an experimental reality, summarizing recent liquid-phase electron microscopy studies of whole cells, assembling peptides, and even individual proteins. Successes and challenges are discussed, as well as strategies to maximize the amount of accessible information and minimize the impact of the electron beam. We conclude with an outlook on the potential of liquid-phase electron microscopy to provide new insight into the rich and functional dynamics occurring in biological systems at the microscopic to molecular level.


Clinical profile and long-term outcome of the first seizures in children.
Seizures are one of the most common causes of pediatric admissions to hospitals in children. This study aims to identify the clinical profile and outcome of first seizures in children. Children who presented to the pediatric neurology outpatient clinic and pediatric emergency service with a first-time seizure and aged one month through 18 years old were enrolled to the study. At the time of the study, enrolled children were categorized into three study groups according to seizure characteristics: febrile seizure, nonfebrile-provoked seizure and, unprovoked seizure. The study group consisted of 138 children. Of the 138 patients, 60 (43%) had febrile first seizures, 23 (17%) had nonfebrile-provoked first seizures, and 55 (40%) had unprovoked first seizures. The patients did not experience the recurrence of a seizure by the treatment of underlying cause at the eighteenth month and the eighth year follow-up in the nonfebrile-provoked seizure group. Among the children admitted for unprovoked first seizures, 33 (60%) patients had seizure recurrence during 18 month follow-up and 36 (82%) patients had seizure recurrence during eight year follow-up. Seizure recurrence rate was statistically higher in patients with abnormal EEG and cranial MRI findings in the unprovoked seizure group (p < .05). The patients with provoked first seizure did not develop epilepsy during eight year follow-up. However, 36 patients with unprovoked seizures were diagnosed with epilepsy during eight year follow-up. It is essential to determine the causes of the seizures and treat the condition.


AL amyloidosis associated with IgM paraproteinemia: clinical profile and treatment outcome.
AL amyloidosis associated with immunoglobulin M (IgM) paraproteinemia is rare. We report 103 consecutive such patients evaluated at the National Amyloidosis Centre (London, United Kingdom) between 1988 and 2006. Renal, cardiac, and lymph node amyloid was present in 53%, 35%, and 21% of patients, respectively, at presentation and 2 or more organs were involved in 54%. Seventy-three percent had an abnormal bone marrow infiltrate (lymphoid in 87%). The median IgM paraprotein was 8 g/L and serum free light chain (FLC) ratio was abnormal in 77 (88%) of 87. The abnormal FLC component was more than 100 mg/L in only 31% cases. Thirty-two percent achieved a partial hematologic response to treatment with no complete responders, and there appeared to be a greater response to combination regimens than single-agent oral alkylators (59% vs 20%, respectively; P = .003). Four achieved amyloidotic organ responses; organ function remained stable in 68%. None with lymph node involvement showed nodal improvement. Median overall survival was 49 months. AL amyloidosis with IgM paraproteinemia represents a distinctive subset of patients with AL amyloidosis who have a wider variety of underlying clonal disorders (often lymphoid) than AL in general, have low-level FLC abnormality, and should be treated with appropriately tailored chemotherapeutic regimens for the underlying clonal disorder.


A Closed-Form Solution to Retinex with Nonlocal Texture Constraints.
We propose a method for intrinsic image decomposition based on retinex theory and texture analysis. While most previous methods approach this problem by analyzing local gradient properties, our technique additionally identifies distant pixels with the same reflectance through texture analysis, and uses these nonlocal reflectance constraints to significantly reduce ambiguity in decomposition. We formulate the decomposition problem as the minimization of a quadratic function which incorporates both the retinex constraint and our nonlocal texture constraint. This optimization can be solved in closed form with the standard conjugate gradient algorithm. Extensive experimentation with comparisons to previous techniques validate our method in terms of both decomposition accuracy and runtime efficiency.


Dorsal column stimulation: optimization of application.
Results are reviewed of dorsal column electrical stimulation for chronic pain in a series of 80 patients. In the light of the experience gained from this series, indications and patient suitability for this form of therapy are discussed. Complications occurring after implantation are tabulated.


The relationship between clients' depression etiological beliefs and psychotherapy orientation preferences, expectations, and credibility beliefs.
The purpose of this study was to examine the relationship between clients' etiological beliefs for depression and treatment preferences, credibility beliefs, and outcome expectations for five different depression treatments-behavioral activation, cognitive therapy, interpersonal psychotherapy, pharmacotherapy, and psychodynamic psychotherapy. Adult psychotherapy clients (N = 98) were asked to complete an online survey that included the Reasons for Depression Questionnaire, a brief description of each of the five treatment options, and credibility, expectancy, and preference questions for each option. On average, the participating clients rated pharmacotherapy as significantly less credible, having a lower likelihood of success, and being less preferred than the four types of psychotherapy. In general, interpersonal psychotherapy was also rated more negatively than the other types of psychotherapy. However, these findings depended somewhat on whether the participating client was personally experiencing depression. Credibility beliefs, outcome expectations, and preferences for pharmacotherapy were positively associated with biological beliefs for depression; however, the other hypothesized relationships between etiological beliefs and treatment attitudes were not supported. Although the study is limited based on the specific sample and treatment descriptions that were used, the results may still have implications for psychotherapy research, training, and practice. (PsycINFO Database Record


Fluoride selectively stimulates Na-dependent phosphate transport in osteoblast-like cells.
The influence of fluoride (F) on the transport of Pi was investigated in the osteoblast-like cell line UMR-106. Exposure of cells to F induced a dose-related stimulation of the Na-coupled Pi transport. Pi transport was significantly increased 6 h after 1 mM F incubation, with maximal response observed at 24 h (F 38.0 +/- 2.3, vehicle 19.8 +/- 1.2 pmol.micrograms DNA-1.4 min-1; P less than 0.001). Na-dependent alanine transport was not changed by F. The selective effect of F on Pi transport was not associated with changes in adenosine 3',5'-cyclic monophosphate, cell proliferation, or alkaline phosphatase activity. However, it was completely blunted by inhibiting translational processes with cycloheximide. Furthermore, F enhanced the stimulatory effect on Pi transport of various mitogens such as fetal calf serum, insulin, and insulin-like growth factor I. In conclusion, F can selectively enhance the activity of the Pi transport system present in the plasma membrane of UMR 106 osteoblast-like cells by a mechanism that probably involves newly synthetized proteins.


Development of a contagious ecthyma vaccine for goats.
To identify a strain of contagious ecthyma virus from goats that possesses the appropriate characteristics for an effective vaccine for goats. 25 goat kids used for vaccine development and 100 goat kids used for evaluation of vaccine efficacy. 5 strains of contagious ecthyma virus were tested in a vaccination-challenge study to identify the best strain to be the seed strain for a contagious ecthyma vaccine. The vaccine derived from the chosen viral stain was tested at 2 concentrations for efficacy in a vaccination-challenge study. 2 of 5 viral strains induced moderate to severe scabs following infection, and 3 viral strains protected the goats from wild-type virus challenge following vaccination. Viral strain 47CE was selected as the seed source for the production of a contagious ecthyma vaccine because of the larger vaccine-to-challenge scab formation ratio. Vaccine 47CE protected all goat kids (48/48) following challenge with the wild-type contagious ecthyma virus; all goat kids (32/32) in the control group had scab formation following challenge with the wild-type contagious ecthyma virus, which indicated no protection following administration of vaccine diluent. A vaccine containing a caprine strain of contagious ecthyma virus used in goats appeared to provide the characteristics needed for an effective vaccine, including good scab production and protection from wild-type infection. This vaccine may potentially provide better protection for goats from contagious ecthyma than currently available vaccines labeled for sheep.


Neutral and selection-driven decay of sexual traits in asexual stick insects.
Environmental shifts and lifestyle changes may result in formerly adaptive traits becoming non-functional or maladaptive. The subsequent decay of such traits highlights the importance of natural selection for adaptations, yet its causes have rarely been investigated. To study the fate of formerly adaptive traits after lifestyle changes, we evaluated sexual traits in five independently derived asexual lineages, including traits that are specific to males and therefore not exposed to selection. At least four of the asexual lineages retained the capacity to produce males that display normal courtship behaviours and are able to fertilize eggs of females from related sexual species. The maintenance of male traits may stem from pleiotropy, or from these traits only regressing via drift, which may require millions of years to generate phenotypic effects. By contrast, we found parallel decay of sexual traits in females. Asexual females produced altered airborne and contact signals, had modified sperm storage organs, and lost the ability to fertilize their eggs, impeding reversals to sexual reproduction. Female sexual traits were decayed even in recently derived asexuals, suggesting that trait changes following the evolution of asexuality, when they occur, proceed rapidly and are driven by selective processes rather than drift.


Herbivores increase the global availability of nutrients over millions of years.
Can the presence of herbivores increase global nutrient availability? Animals disperse vital nutrients through ecosystems, increasing the spatial availability of these nutrients. Large herbivores are especially important for the dispersal of vital nutrients due to their long food passage times and day ranges, and large herbivores from past periods (the Pleistocene) may have increased nutrient concentrations on the continental scale. However, such results have been demonstrated theoretically but not yet empirically. Models suggest that the Pennsylvanian subperiod (323-299 million years ago), with no tetrapod terrestrial herbivores, would have had fewer, less-well-distributed nutrients than the Cretaceous period (145-66 million years ago), with the largest terrestrial herbivores ever-the sauropods. Here, I show that these models are supported empirically by remnant plant material (coal deposits) from the Cretaceous (N = 680), which had significantly (P < 0.00001) increased concentrations (136%) and decreased spatial heterogeneity (22%) of plant-important rock-derived nutrients compared with the Pennsylvanian subperiod (N = 4,996). Non-biotic physical processes, such as weathering rates, cannot account for such differences, because aluminium-a nutrient not important for plants and animals, but weathered in a similar manner to the above elements-showed no significant difference between the two periods, suggesting that these large changes were driven by plant-herbivore interactions. Populations of large wild herbivores are currently at historical lows; therefore, we are potentially losing a key ecosystem service.


Complete Genome Sequences of Isolates of Enterococcus faecium Sequence Type 117, a Globally Disseminated Multidrug-Resistant Clone.
The emergence of nosocomial infections by multidrug-resistant sequence type 117 (ST117) Enterococcus faecium has been reported in several European countries. ST117 has been detected in Spanish hospitals as one of the main causes of bloodstream infections. We analyzed genome variations of ST117 strains isolated in Madrid and describe the first ST117 closed genome sequences.


Gold-Embedded Hollow Silica Nanogolf Balls for Imaging and Photothermal Therapy.
Hybrid nanocarriers with multifunctional properties have wide therapeutic and diagnostic applications. We have constructed hollow silica nanogolf balls (HGBs) and gold-embedded hollow silica nanogolf balls (Au@SiO2 HGBs) using the layer-by-layer approach on a symmetric polystyrene (PS) Janus template; the template consists of smaller PS spheres attached to an oppositely charged large PS core. ζ Potential measurement supports the electric force-based template-assisted synthesis mechanism. Electron microscopy, UV-vis, and near-infrared (NIR) spectroscopy show that HGBs or Au@SiO2 HGBs are composed of a porous silica shell with an optional dense layer of gold nanoparticles embedded in the silica shell. To visualize their cellular uptake and imaging potential, Au@SiO2 HGBs were loaded with quantum dots (QDs). Confocal fluorescent microscopy and atomic force microscopy imaging show reliable endocytosis of QD-loaded Au@SiO2 HGBs in adherent HeLa cells and circulating red blood cells (RBCs). Surface-enhanced Raman spectroscopy of Au@SiO2 HGBs in RBC cells show enhanced intensity of the Raman signal specific to the RBCs' membrane specific spectral markers. Au@SiO2 HGBs show localized surface plasmon resonance and heat-induced HeLa cell death in the NIR range. These hybrid golf ball nanocarriers would have broad applications in personalized nanomedicine ranging from in vivo imaging to photothermal therapy.


A synthetic analysis of the Saccharomyces cerevisiae stress sensor Mid2p, and identification of a Mid2p-interacting protein, Zeo1p, that modulates the PKC1-MPK1 cell integrity pathway.
Mid2p is a plasma membrane protein that functions in Saccharomyces cerevisiae as a sensor of cell wall stress, activating the PKC1-MPK1 cell integrity pathway via the small GTPase Rho1p during exposure to mating pheromone, calcofluor white, and heat. To examine Mid2p signalling, a global synthetic interaction analysis of a mid2 mutant was performed; this identified 11 interacting genes. These include WSC1 and ROM2, upstream elements in cell integrity pathway signalling, and FKS1 and SMI1, required for 1,3-beta-glucan synthesis. These synthetic interactions indicate that the Wsc1p sensor acts through Rom2p to activate the Fks1p glucan synthase in a Mid2p-independent way. To further explore Mid2p signalling a two-hybrid screen was done using the cytoplasmic tail of Mid2p; this identified ZEO1 (YOL109w), encoding a 12 kDa peripheral membrane protein that localizes to the plasma membrane. Disruption of ZEO1 leads to resistance to calcofluor white and to a Mid2p-dependent constitutive phosphorylation of Mpk1p, supporting a role for Zeo1p in the cell integrity pathway. Consistent with this, zeo1-deficient cells suppress the growth defect of mutants in the Rho1p GDP-GTP exchange factor Rom2p, while exacerbating the growth defect of sac7delta mutants at 37 degrees C. In contrast, mid2delta mutants have opposing effects to zeo1delta mutants, being synthetically lethal with rom2delta, and suppressing an 18 degrees C growth defect of sac7delta, while overexpression of MID2 rescues a rom2delta 37 degrees C growth defect. Thus, MID2 and ZEO1 appear to play reciprocal roles in the modulation of the yeast PKC1-MPK1 cell integrity pathway.


Effects of Ligustrazine on Airway Inflammation in A Mouse Model of Neutrophilic Asthma.
To investigate the effects of ligustrazine (LTZ) on airway inflammation in a mouse model of neutrophilic asthma (NA). Forty healthy C57BL/6 female mice were randomly divided into 4 groups using a random number table, including the normal control, NA, LTZ and dexamethasone (DXM) groups, with 10 rats in each group. The NA mice model was established by the method of ovalbumin combined with lipopolysaccharide sensitization. At 0.5 h before each challenge, LTZ and DXM groups were intraperitoneally injected with LTZ (80 mg/kg) or DXM (0.5 mg/kg) for 14 d, respectively, while the other two groups were given the equal volume of normal saline. After last challenge for 24 h, the aerosol inhalation of methacholine was performed and the airway reactivity was measured. The bronchoalveolar lavage fluid (BALF) was collected. The Wright-Giemsa staining was used for total white blood cells and differential counts. The levels of cytokines interleukin (IL)-17 and IL-10 were detected by enzyme-linked immunosorbent assay. The pathological change of lung tissue was observed by hematoxylin eosin staining. The airway responsiveness of the NA group was signifificantly higher than the normal control group (P<0.05), while those in the LTZ and DXM groups were signifificantly lower than the NA group (P<0.05). The neutrophil and eosinophil counts in the LTZ and DXM groups were signifificantly lower than the NA group (P<0.05), and those in the LTZ group were signifificantly lower than the DXM group (P<0.05). There were a large number of peribronchiolar and perivascular inflammatory cells in fifiltration in the NA group. The airway inflflammation in the LTZ and DXM groups were signifificantly alleviated than the NA group. The infifiltration in the LTZ group was signifificantly reduced than the DXM group. Compared with the normal control group, the IL-17 level in BALF was signifificantly increased and the IL-10 level in BALF was signifificantly decreased in the NA group (P<0.05). LTZ and DXM treatment signifificantly decreased IL-17 levels and increased IL-10 levels compared with the NA group (P<0.05), and the changes in the above indices were more signifificant in the LTZ group (P<0.05). LTZ could alleviate the airway inflflammation in the NA mice model through increasing the IL-10 level and decreasing the IL-17 level.


Utility of the harmonic scalpel for laparoscopic partial nephrectomy.
Laparoscopic partial nephrectomy (LPN) remains a technically challenging procedure largely because of the lack of methods for obtaining consistent parenchymal hemostasis. The objective of this study was to determine if the extent of resection influences the ability of the harmonic scalpel to achieve hemostasis and to define the cases in which the harmonic scalpel is appropriate for LPN. Thirty LPNs were performed in a 25-kg domestic pig model. The blunt blade of the laparoscopic harmonic scalpel (LaparoSonic Coagulating Shears; Ethicon Endo-Surgery, Cincinnati, OH) at power level 5 was used to divide the parenchyma. Control of the renal hilar vessels was not obtained. Three standardized types of resections were performed: I = peripheral wedge biopsy; II = upper- or lower-pole nephrectomy; and III = heminephrectomy. Bleeding was graded on a scale from 0 to 4: 0 = no hemostasis; 1 = steady bleeding; 2 = moderate bleeding; 3 = parenchymal oozing; and 4 = dry. Hemostasis grades of 2 or less were clinically significant bleeding necessitating supplemental coagulation. The mean hemostasis scores showed a significant (P < 0.02) trend toward inadequate hemostasis with increasing extent of resection: 3.3 for Type I, 3.0 for Type II, and 2.4 for Type III. The percent of kidneys with grade 2 bleeding or worse was 9% for Type I surgery, 25% for Type II, and 57% for Type III. Successful hemostasis with the harmonic scalpel correlates with the extent of parenchymal resection in the porcine model. Most wedge excisions can be done with the harmonic scalpel alone, whereas larger resections necessitate supplemental coagulation. On the basis of this study, heminephrectomies with the harmonic scalpel are not recommended because of the high incidence of significant hemorrhage.


[Hormonal regulation of mucoprotein secretion by salivary gland cells of Drosophila melanogaster larvae during metamorphosis].
The secretion of mucoproteins by the Drosophila melanogaster salivary gland cells into the gland lumen is associated with the high ecdysone content. Mucoproteins are synthesized but not discharged by these cells in ecdysone-deficient homozygous 1(2)gl larvae, as well as in normal larvae devoid of ecdysone by ligatures and transplantation of salivary gland into the imago abdomen. Ecdysone injections stimulate the mucoprotein secretion into the gland lumen.


Towards an understanding of equine pleuropneumonia: factors relevant for control.
To review relevant literature on factors associated with the development of equine pleuropneumonia. A review of the literature using a range of databases including Current Contents, Medline, ChemAbstracts, Biological Abstracts and CAB and a comprehensive search strategy which involved use of keywords, author and subject category searches. Additional sources included review of articles cited by key accumulated references. Since the early years of this century, many of the "gaps" in our knowledge of the pathogenesis of this disease have been filled. We now know that equine pleuropneumonia results from contamination of the lower respiratory tract with bacteria similar to the normal oropharyngeal microbiota of the horse and that transportation of any mode, especially over long distances (and consequently with no or short rest periods), is the single most important predisposing factor for this disease. This is associated with restraint of horses such that they are unable to lower their heads, which leads to increased opportunity for lower respiratory tract contamination and a reduced opportunity for clearance. Strenuous exercise also results in lower respiratory tract contamination and exercise subsequent to transportation exerts additive detrimental effects on the defenses of the lower respiratory tract. While modern veterinary medicine and surgery have significantly reduced the death rate from pleuropneumonia, horses that develop the disease have a high probability of not returning to their prior use. This underscores the importance of developing the most effective strategies for its prevention.


Deficient cell cycle control in myeloid cells of patients with newly diagnosed chronic myeloid leukemia.
Cell cycle control subsequent to gamma irradiation or growth factor starvation has been studied in immative hematopoietic cells of 19 previously untreated chronic myeloid leukemia (CML) patients in chronic phase compared with 18 normal controls. CD34-positive cells were cultured for seven days in the presence of optimal concentrations of appropriate growth factors. At day 7 of culture both S-phase fraction and differentiation were identical in normal and leukemic cells. In normal cells the proportion of S-phase cells was reduced by irradiation with 500 rad from 40 +/- 3% to 16 +/- 2%. In contrast, in CML cells a reduction of S-phase cells from 35 +/- 2% to 25 +/- 3% was observed. Moreover, irradiated CML cells arrested at a smaller number of cells in G2. Similarly, a significantly higher proportion of CML cells remained in S phase after withdrawal of growth factors. Semiquantitative PCR of p21 (waf1/cip1) induction by gamma irradiation provided no evidence for a major functional deficiency of p53 response to irradiation in these cells. Our results demonstrate an abnormal cell cycle arrest in chronic-phase CML cells both after gamma irradiation and after growth factor removal. This observation might have important implications for understanding the pathogenesis of both hyperplasia of chronic phase and the development of blast crisis in CML. The molecular mechanisms underlying these abnormalities in bcr-abl-positive cells remain to be clarified.


Detection of infectious bronchitis virus.
The detection methods for infectious bronchitis virus (IBV) are reviewed. Advantages and disadvantages of available techniques of IBV detection by virus isolation, antigen or genome detection, and serology are discussed. Factors of influence on the level of success in detection of IBV after a disease outbreak are discussed, as are the possibilities and dangers of strain classification by protectotyping, serotyping, epitope-typing and genotyping.


Visualisation of dCas9 target search in vivo using an open-microscopy framework.
CRISPR-Cas9 is widely used in genomic editing, but the kinetics of target search and its relation to the cellular concentration of Cas9 have remained elusive. Effective target search requires constant screening of the protospacer adjacent motif (PAM) and a 30 ms upper limit for screening was recently found. To further quantify the rapid switching between DNA-bound and freely-diffusing states of dCas9, we developed an open-microscopy framework, the miCube, and introduce Monte-Carlo diffusion distribution analysis (MC-DDA). Our analysis reveals that dCas9 is screening PAMs 40% of the time in Gram-positive Lactoccous lactis, averaging 17 ± 4 ms per binding event. Using heterogeneous dCas9 expression, we determine the number of cellular target-containing plasmids and derive the copy number dependent Cas9 cleavage. Furthermore, we show that dCas9 is not irreversibly bound to target sites but can still interfere with plasmid replication. Taken together, our quantitative data facilitates further optimization of the CRISPR-Cas toolbox.


Improved Microbial Community Characterization of 16S rRNA via Metagenome Hybridization Capture Enrichment.
Environmental microbial diversity is often investigated from a molecular perspective using 16S ribosomal RNA (rRNA) gene amplicons and shotgun metagenomics. While amplicon methods are fast, low-cost, and have curated reference databases, they can suffer from amplification bias and are limited in genomic scope. In contrast, shotgun metagenomic methods sample more genomic regions with fewer sequence acquisition biases, but are much more expensive (even with moderate sequencing depth) and computationally challenging. Here, we develop a set of 16S rRNA sequence capture baits that offer a potential middle ground with the advantages from both approaches for investigating microbial communities. These baits cover the diversity of all 16S rRNA sequences available in the Greengenes (v. 13.5) database, with no sequence having <78% sequence identity to at least one bait for all segments of 16S. The use of our baits provide comparable results to 16S amplicon libraries and shotgun metagenomic libraries when assigning taxonomic units from 16S sequences within the metagenomic reads. We demonstrate that 16S rRNA capture baits can be used on a range of microbial samples (i.e., mock communities and rodent fecal samples) to increase the proportion of 16S rRNA sequences (average > 400-fold) and decrease analysis time to obtain consistent community assessments. Furthermore, our study reveals that bioinformatic methods used to analyze sequencing data may have a greater influence on estimates of community composition than library preparation method used, likely due in part to the extent and curation of the reference databases considered. Thus, enriching existing aliquots of shotgun metagenomic libraries and obtaining modest numbers of reads from them offers an efficient orthogonal method for assessment of bacterial community composition.


Enzyme-instructed assembly of a cholesterol conjugate promotes pro-inflammatory macrophages and induces apoptosis of cancer cells.
The combination of immunotherapy and chemotherapy has shown great potential for treating solid tumors. Although various combination therapy regimens have been demonstrated to be effective, their implementation in practice always needs separate drugs or modalities, which in turn requires specific and complicated operation procedures. Here, we describe the use of enzyme-instructed self-assembly (EISA) of a phosphotyrosine-cholesterol (PTC) conjugate to combine immune microenvironment modulation and chemotherapy. The enzymatic dephosphorylation of PTC results in the assembly of its derivatives inside and outside the cells, which leads to repolarization of the macrophage phenotype and direct death of cancer cells. The generation of reactive oxygen species and actin disturbance induced by the assemblies has been verified relative to the macrophage repolarization. We also demonstrate its dual repolarization and inhibition effects in vivo using an ovarian cancer bearing immunocompetent murine model. The EISA of cholesterol conjugates is an effective yet simple approach for the combination of tumor microenvironment immunomodulation and chemotherapy.


Taming quantum interference in single molecule junctions: induction and resonance are key.
We have joined two fundamental concepts of organic chemistry to provide a deep, yet intuitive, understanding of how side groups influence destructive quantum interference (DQI) in the transport through conjugated molecules. Using density functional theory combined with Green's function techniques, and employing tight-binding models in which all the π-systems are considered, we elucidate the separate roles of bond-resonance and induction in tuning DQI. We show that the position of the anti-resonances produced by DQI is sensitive to the number of side groups, but not in a simple additive way. Instead, addition of multiple groups results in a weaker overall contribution per group, and this can be understood using a straight forward graphical analysis. Furthermore, we show that additional fine tuning of DQI is possible via attachment of a chain of atoms to a second site around the ring. DQI is controlled by modifying the length of the chain, thus providing exquisite control over the anti-resonance position. This insight provides chemists with a large number of options to tune DQI for unprecedented device optimization.


Aztreonam versus gentamicin for short-term prophylaxis in biliary and gastric surgery.
Short-term antibiotic prophylaxis was studied in 80 patients undergoing biliary or gastric surgery. The patients were randomized to receive 1 g of aztreonam or 80 mg of gentamicin intravenously 30 minutes before surgery and 8 and 16 hours after surgery. Of samples taken from the abdominal cavity for bacteriologic study, 53% were culture positive. Wound infections developed in two (4.5%) of 44 patients receiving aztreonam and in seven (19.4%) of 36 patients treated with gentamicin. Staphylococcus epidermidis and Enterobacter species were isolated from sites of wound infection in the aztreonam group; Escherichia coli (two isolates), Pseudomonas aeruginosa (two isolates), Enterobacter species, Klebsiella species, Enterococcus faecalis, and Aeromonas hydrophila were isolated from the gentamicin group. Our data indicate that aztreonam is safe and effective for the prevention of infections following biliary and gastric surgery.


Uptake of copper ion by activated sludge and its bacterial community variation analyzed by 16S rDNA.
The effect and uptake of copper ion on SBR (sequence batch reactor) biological treatment system was studied. Special nutrient and powder activated carbon (PAC) additive were tested as uptake stimulation technique. Results showed that copper ion had higher effect on unacclimated activated sludge system than on acclimated one. The special nutrient adding could enhance the uptake of copper significantly, while PAC adding could improve the sludge settling and decrease the turbidity of effluent. The variation of bacterial community analyzed by 16S rDNA method showed the acclimation of copper could increase copper resistance species, and excess accumulation could cause some species diminish. It was confirmed that acclimation could improve the resistance and uptake ability of microorganism to heavy metal.


Exercise protects against myocardial ischemia-reperfusion injury via stimulation of β(3)-adrenergic receptors and increased nitric oxide signaling: role of nitrite and nitrosothiols.
Exercise training confers sustainable protection against ischemia-reperfusion injury in animal models and has been associated with improved survival following a heart attack in humans. It is still unclear how exercise protects the heart, but it is apparent that endothelial nitric oxide synthase (eNOS) and nitric oxide (NO) play a role. To determine the role of β(3)-adrenergic receptors (β(3)-ARs), eNOS activation, and NO metabolites (nitrite and nitrosothiols) in the sustained cardioprotective effects of exercise. Here we show that voluntary exercise reduces myocardial injury in mice following a 4-week training period and that these protective effects can be sustained for at least 1 week following the cessation of the training. The sustained cardioprotective effects of exercise are mediated by alterations in the phosphorylation status of eNOS (increase in serine 1177 and decrease in threonine 495), leading to an increase in NO generation and storage of NO metabolites (nitrite and nitrosothiols) in the heart. Further evidence revealed that the alterations in eNOS phosphorylation status and NO generation were mediated by β(3)-AR stimulation and that in response to exercise a deficiency of β(3)-ARs leads to an exacerbation of myocardial infarction following ischemia-reperfusion injury. Our findings clearly demonstrate that exercise protects the heart against myocardial ischemia-reperfusion injury by stimulation of β(3)-ARs and increased cardiac storage of nitric oxide metabolites (ie, nitrite and nitrosothiols).


The Vulvovaginal Symptoms Questionnaire: a questionnaire for measuring vulvovaginal symptoms in postmenopausal women.
This study aims to develop a Vulvovaginal Symptoms Questionnaire (VSQ) to determine the symptoms, emotions, life impact, and sexual impact of vulvovaginal symptoms on postmenopausal women. We developed a questionnaire that focuses on vulvovaginal symptoms based on modifications to Skindex-16, a validated questionnaire for measuring the impact of skin disease. We recruited postmenopausal women seeking routine gynecologic care to test the psychometric properties of the VSQ. Test-retest reliability was assessed 2 to 4 weeks after their initial recruitment and measured using intraclass coefficients. Four distinct a priori scales of the VSQ were developed: symptoms, emotions, life impact, and sexual impact. Confirmatory factor analysis was performed to verify the four a priori scales by evaluating the goodness of fit of a final confirmatory factor analysis model. The internal consistency of the scales was assessed by calculating Cronbach's α coefficients. The VSQ is a 21-item written questionnaire with four scales: symptoms, emotions, life impact, and sexual impact. One hundred twenty postmenopausal women participated in the psychometric validation of the VSQ. The test-retest reliability measured by intraclass coefficients was 0.75, 0.60, 0.55, and 0.65 for the symptoms, emotions, life impact, and sexual impact scales, respectively. The goodness of fit of the confirmatory factor response model was confirmed. The Cronbach's α coefficients were 0.76, 0.87, 0.83, and 0.82 for the respective scales. The VSQ is a reliable and internally consistent instrument for measuring vulvovaginal symptoms in postmenopausal women.


Intestinal secretory defects and dwarfism in mice lacking cGMP-dependent protein kinase II.
Cyclic guanosine 3',5'-monophosphate (cGMP)-dependent protein kinases (cGKs) mediate cellular signaling induced by nitric oxide and cGMP. Mice deficient in the type II cGK were resistant to Escherichia coli STa, an enterotoxin that stimulates cGMP accumulation and intestinal fluid secretion. The cGKII-deficient mice also developed dwarfism that was caused by a severe defect in endochondral ossification at the growth plates. These results indicate that cGKII plays a central role in diverse physiological processes.


Association of symptomatic benign prostatic hyperplasia and prostate cancer: results from the prostate cancer prevention trial.
This study examined the association between symptomatic benign prostatic hyperplasia (BPH) and prostate cancer risk in 5,068 placebo-arm participants enrolled in the Prostate Cancer Prevention Trial (1993-2003). These data include 1,225 men whose cancer was detected during the 7-year trial--556 detected for cause (following abnormal prostate-specific antigen or digital rectal examination) and 669 detected not for cause (without indication), as well as 3,843 men who had biopsy-proven absence of prostate cancer at the trial end. Symptomatic BPH was assessed hierarchically as self-report of surgical or medical treatment, moderately severe symptoms (International Prostate Symptom Score >14), or physician diagnosis, and analyses were completed by BPH status at baseline (prevalent) or BPH prior to cancer diagnosis or study end (prevalent plus incident). Controlled for age, race, and body mass index, neither prevalent (risk ratio = 1.03, 95% confidence interval: 0.92, 1.14) nor prevalent plus incident (risk ratio = 0.96, 95% confidence interval: 0.87, 1.06) symptomatic BPH was associated with prostate cancer risk. This lack of association was consistent across subgroups defined by type of BPH-defining event (treatment, symptoms, or physician diagnosis), prompt for prostate cancer diagnosis, and prostate cancer grade. This study provides the strongest evidence to date that BPH does not increase the risk of prostate cancer.


Targeting ticagrelor: a novel therapy for emergency reversal.
Newer P2Y12 inhibitors are prescribed in place of clopidogrel for patients with acute coronary syndrome (ACS) and are associated with significant bleeding risks. Currently, limited options exist for the management of life-threatening bleeding or acute reversal for patients on P2Y12 inhibitor therapy, specifically ticagrelor. Various interventions, including platelet transfusion and desmopressin, have been studied for ticagrelor reversal demonstrating limited success. PB2452 is a novel monoclonal antibody which binds to both ticagrelor and its active metabolite resulting in a rapid return of platelet aggregation. PB2452 has been studied in animal models and, most recently, in a Phase I trial in healthy volunteers. In animal models, PB2452 displayed rapid reversal of ticagrelor and its metabolites and return to near normal levels of platelet aggregation within 60 min. In healthy human volunteers, cohorts that received higher dose bolus and infusions of PB2452 over 12-16 h resulted in maximal and sustained reversal of ticagrelor inhibition of platelet aggregation. While it is currently not US Food and Drug Administration approved, future Phase 2 and 3 studies are currently underway that may lead to new directions for patients on ticagrelor therapy who require urgent reversal.


Conceptual analysis of compliance.
Compliance has been studied from a wide range of scientific perspectives including medicine, nursing, psychology and health economics. There is no agreement regarding a commonly accepted definition. Lack of consistency in the definition and measurement of compliance is a major problem in research which becomes more complicated in an international study. The response to the confusion over the term compliance has been to suggest and use alternative terms such as adherence, co-operation, mutuality and therapeutic alliance. These terms are ill-defined and often are used as synonyms. The purpose of this paper is to analyse definitions of the concept of compliance. Abstracts from MEDLINE have been analysed in order to identify the types of compliance research that have been carried out.


Inhibition of microsomal epoxide hydrolases by ureas, amides, and amines.
The microsomal epoxide hydrolase (mEH) plays a significant role in the metabolism of xenobiotics such as polyaromatic toxicants. Additionally, polymorphism studies have underlined a potential role of this enzyme in relation to several diseases, such as emphysema, spontaneous abortion, and several forms of cancer. To provide new tools for studying the function of mEH, inhibition of this enzyme was investigated. Inhibition of recombinant rat and human mEH was achieved using primary ureas, amides, and amines. Several of these compounds are more potent than previously published inhibitors. Elaidamide, the most potent inhibitor that is obtained, has a K(i) of 70 nM for recombinant rat mEH. This compound interacts with the enzyme forming a noncovalent complex, and blocks substrate turnover through an apparent mix of competitive and noncompetitive inhibition kinetics. Furthermore, in insect cell cultures expressing rat mEH, elaidamide enhances the toxicity effects of epoxide-containing xenobiotics. These inhibitors could be valuable tools for investigating the physiological and toxicological roles of mEH.


High fluoroquinolone resistance proportions among multidrug-resistant tuberculosis driven by dominant L2 Mycobacterium tuberculosis clones in the Mumbai Metropolitan Region.
Multidrug-resistant (MDR) Mycobacterium tuberculosis complex (MTBC) strains are a serious health problem in India, also contributing to one-fourth of the global MDR tuberculosis (TB) burden. About 36% of the MDR MTBC strains are reported fluoroquinolone (FQ) resistant leading to high pre-extensively drug-resistant (pre-XDR) and XDR-TB (further resistance against bedaquiline and/or linezolid) rates. Still, factors driving the MDR/pre-XDR epidemic in India are not well defined. In a retrospective study, we analyzed 1852 consecutive MTBC strains obtained from patients from a tertiary care hospital laboratory in Mumbai by whole genome sequencing (WGS). Univariate and multivariate statistics was used to investigate factors associated with pre-XDR. Core genome multi locus sequence typing, time scaled haplotypic density (THD) method and homoplasy analysis were used to analyze epidemiological success, and positive selection in different strain groups, respectively. In total, 1016 MTBC strains were MDR, out of which 703 (69.2%) were pre-XDR and 45 (4.4%) were XDR. Cluster rates were high among MDR (57.8%) and pre-XDR/XDR (79%) strains with three dominant L2 (Beijing) strain clusters (Cl 1-3) representing half of the pre-XDR and 40% of the XDR-TB cases. L2 strains were associated with pre-XDR/XDR-TB (P < 0.001) and, particularly Cl 1-3 strains, had high first-line and FQ resistance rates (81.6-90.6%). Epidemic success analysis using THD showed that L2 strains outperformed L1, L3, and L4 strains in short- and long-term time scales. More importantly, L2 MDR and MDR + strains had higher THD success indices than their not-MDR counterparts. Overall, compensatory mutation rates were highest in L2 strains and positive selection was detected in genes of L2 strains associated with drug tolerance (prpB and ppsA) and virulence (Rv2828c). Compensatory mutations in L2 strains were associated with a threefold increase of THD indices, suggesting improved transmissibility. Our data indicate a drastic increase of FQ resistance, as well as emerging bedaquiline resistance which endangers the success of newly endorsed MDR-TB treatment regimens. Rapid changes in treatment and control strategies are required to contain transmission of highly successful pre-XDR L2 strains in the Mumbai Metropolitan region but presumably also India-wide.


High rates of lateral gene transfer are not due to false diagnosis of gene absence.
Methods for assessing gene presence and absence have been widely used to study bacterial genome evolution. A recent report by Zhaxybayeva et al. [Zhaxybayeva, O., Nesbo, C. L., and Doolittle, W. F., 2007. Systematic overestimation of gene gain through false diagnosis of gene absence. Genome. Biol. 8, 402] suggests that false diagnosis of gene absence or the presence of undetected truncated genes leads to a systematic overestimation of gene gain. Here (1) we argue that these annotation errors can cause more complicated effects and are not necessarily systematic, (2) we argue that current annotations (supplemented with BLAST searches) are the best way to consistently score gene presence/absence and (3) that genome wide estimates of gene gain/loss are not strongly affected by small differences in gene annotations but that the number of related gene families is strongly affected. We have estimated the rates of gene insertions/deletions using a variety of cutoff thresholds and match lengths as a way in which to alter the recognition of genes and gene fragments. The results reveal that different cutoffs for match length only cause a small variation of the estimated insertion/deletion rates. The rates of gene insertions/deletions on recent branches remain relatively high regardless of the thresholds for match length. Lastly (4), the dynamic process of gene truncation needs to be further considered in genome comparison studies. The data presented suggest that gene truncation tends to take place preferentially in recently transferred genes, which supports a fast turnover of recent laterally transferred genes. The presence of truncated genes or false diagnosis of gene absence therefore does not significantly affect the estimation of gene insertions/deletions rates, but there are several other factors that bias the results toward an under-estimation of the rate of gene insertion/deletion. All of these factors need to be considered.


L19-IL2 Immunocytokine in Combination with the Anti-Syndecan-1 46F2SIP Antibody Format: A New Targeted Treatment Approach in an Ovarian Carcinoma Model.
Epithelial ovarian cancer (EOC) is the fifth most common cancer affecting the female population. At present, different targeted treatment approaches may improve currently employed therapies leading either to the delay of tumor recurrence or to disease stabilization. In this study we show that syndecan-1 (SDC1) and tumor angiogenic-associated B-fibronectin isoform (B-FN) are involved in EOC progression and we describe the prominent role of SDC1 in the vasculogenic mimicry (VM) process. We also investigate a possible employment of L19-IL2, an immunocytokine specific for B-FN, and anti-SDC1 46F2SIP (small immuno protein) antibody in combination therapy in a human ovarian carcinoma model. A tumor growth reduction of 78% was obtained in the 46F2SIP/L19-IL2-treated group compared to the control group. We observed that combined treatment was effective in modulation of epithelial-mesenchymal transition (EMT) markers, loss of stemness properties of tumor cells, and in alleviating hypoxia. These effects correlated with reduction of VM structures in tumors from treated mice. Interestingly, the improved pericyte coverage in vascular structures suggested that combined therapy could be efficacious in induction of vessel normalization. These data could pave the way for a possible use of L19-IL2 combined with 46F2SIP antibody as a novel therapeutic strategy in EOC.


[The verification possibilities of immunochips under low content of antibodies to Core-antigen of virus of hepatitis C].
The detection of antibodies to Core-antigen of virus of hepatitis C in test-systems for solid-phase immune-enzyme analysis with low optical density can be a result not only of true availability of antibodies but an effect of nonspecific reaction of blood serum. The diagnostic possibilities of immunochips to be used in immune-enzyme analysis for verification of availability of markers of viral hepatitis C were investigated in conditions of low positive reaction of blood serum to core-antigen. It is established that immunochips and immunoblots have similar specificity concerning detection of antibodies to Core-antigen. At that, in immunochips antibodies to nonstructural antigens of virus of hepatitis C were additionally detected in more than 90% of samples.


Predicting the influence of multiple components on microbial inhibition using a logistic response model--a novel approach.
There are several synergistic methods available. However, there is a vast discrepancy in the interpretation of the synergistic results. Also, these synergistic methods do not assess the influence the tested components (drugs, plant and natural extracts), have upon one another, when more than two components are combined. A modified checkerboard method was used to evaluate the synergistic potential of Heteropyxis natalensis, Melaleuca alternifolia, Mentha piperita and the green tea extract known as TEAVIGO™. The synergistic combination was tested against the oral pathogens, Streptococcus mutans, Prevotella intermedia and Candida albicans. Inhibition data obtained from the checkerboard method, in the form of binary code, was used to compute a logistic response model with statistically significant results (p < 0.05). This information was used to construct a novel predictive inhibition model. Based on the predictive inhibition model for each microorganism, the oral pathogens tested were successfully inhibited (at 100% probability) with their respective synergistic combinations. The predictive inhibition model also provided information on the influence that different components have upon one another, and on the overall probability of inhibition. Using the logistic response model negates the need to 'calculate' synergism as the results are statistically significant. In successfully determining the influence multiple components have upon one another and their effect on microbial inhibition, a novel predictive model was established. This ability to screen multiple components may have far reaching effects in ethnopharmacology, agriculture and pharmaceuticals.


Loss-of-function mutations in MC4R are very rare in the Greek severely obese adult population.
Melanocortin-4 receptor (MC4R) loss-of-function mutations are the commonest genetic cause of human monogenic obesity, so far. The contribution of MC4R coding mutations to severe obesity in the high-obesity prone Greek population has not been investigated to date. We determined the MC4R coding sequence of 510 obese and 469 lean control subjects of Greek origin, and we estimated the prevalence and the penetrance on obesity of MC4R loss-of-function mutations. The functional impact of novel nonsynonymous variants detected was investigated in vitro. We found two novel synonymous mutations (L23L and I102I), four nonsynonymous mutations (T112M, S127L, N274S, and S295L), and two polymorphisms (V103I and I251L) previously described in literature. We also detected a novel mutation (L207V) in a severely obese 69-year-old female patient, although the mutation did not cosegregate with obesity in the corresponding pedigree and had no functional consequences on MC4R protein function. Loss-of-function mutations represented 75% of all nonsynonymous rare mutations identified among lean carriers and only 25% among obese subjects (P = 0.0001). The prevalence of loss-of-function mutations was lower in the obese group than in lean control subjects (0.20 vs. 0.64%) but this difference was not significant. Therefore, the estimated penetrance of deleterious MC4R mutations was very low (6.3%) in heterozygous Greek carriers of MC4R loss-of-function mutations. Our data suggest that MC4R loss-of-function mutations are rare in the Greek population. MC4R genetic deficiency is unlikely to explain the high propensity to develop severe obesity in this specific population.


Seeking refuge in the present moment: A qualitatively refined model of dispositional mindfulness, minority stress, and psychosocial health among Latino/a sexual minorities and their families.
Racial/ethnic and sexual minorities may experience excess stress from social prejudice and structural disadvantage, which is associated with family relationship problems and poorer psychosocial health. There is increasing evidence that certain positive psychological processes can attenuate these negative outcomes at the individual and interpersonal levels; however, the mechanisms of these effects remain poorly understood. Based on previous research and two extant conceptual frameworks, we constructed a preliminary model of how dispositional mindfulness operated in the context of minority stress among Latino/a sexual minority young adults and their families. We then conducted semi-structured interviews in a life history format with 21 Latino/a sexual minority young adults and 15 family members to test our preliminary model and refine it. We mixed content analysis and grounded theory techniques, whereby we initially used deductive coding with pre-established codes based on our preliminary model, and subsequently used inductive coding to account for novel categories in the interview data that could not be explained by the preliminary model. The refined model revealed pathways by which five constructs of dispositional mindfulness (act with awareness, observe, describe, non-judge, non-react) buffered negative effects of minority stress on psychosocial health in young adults, and were qualitatively associated with compassion, acceptance of sexual minorities, and closeness among family members. Young adults reporting deficiencies in mindfulness reported greater suffering and negative coping related to minority stress. Our model provides a framework to explore modes of resilience and adaptation to minority stress, and offers a basis for further refinement in other affected populations.


Prevalence rates of youth gambling problems: are the current rates inflated?
While there is a general consensus in the literature that it is common for youth to gamble, considerable variability in the reported prevalence rates of youth problem gambling has been found. More recently, issues concerning the possible overestimation of these rates have been raised. Arguments underlying the proposition that problem gambling rates for youth are inflated are examined. It is acknowledged that more rigorous research is required, including the need for the development and refinement of current adolescent instruments and screening tools, agreement upon a gold standard criterion for adolescent problem gambling, and clarity of nomenclature issues. The advancement of scientific knowledge concerning the underlying risk factors associated with the onset and course of youth gambling involvement and the role of effective adolescent prevention and treatment programs will require these fundamental research questions to be addressed.


Non-Cationic Proteins Are Associated with HIV Neutralizing Activity in Genital Secretions of Female Sex Workers.
Cationic proteins found in cervicovaginal secretions (CVS) are known to contribute to the early antiviral immune response against HIV-infection in vitro. We here aimed to define additional antiviral factors that are over-expressed in CVS from female sex workers at high risk of infection. CVS were collected from Kenyan HIV-seronegative (n = 34) and HIV-seropositive (n = 12) female sex workers, and were compared with those from HIV-seronegative low-risk women (n = 12). The highly exposed seronegative (HESN) sex workers were further divided into those with less (n = 22) or more (n = 12) than three years of documented sex work. Cationic protein-depleted CVS were assessed for HIV-neutralizing activity by a PBMC-based HIV-neutralizing assay, and then characterized by proteomics. HIV neutralizing activity was detected in all unprocessed CVS, however only CVS from the female sex worker groups maintained its HIV neutralizing activity after cationic protein-depletion. Differentially abundant proteins were identified in the cationic protein-depleted secretions including 26, 42, and 11 in the HESN>3 yr, HESN<3 yr, and HIV-positive groups, respectively. Gene ontology placed these proteins into functional categories including proteolysis, oxidation-reduction, and epidermal development. The proteins identified in this study include proteins previously associated with the HESN phenotype in other cohorts as well as novel proteins not yet associated with anti-HIV activities. While cationic proteins appear to contribute to the majority of the intrinsic HIV neutralizing activity in the CVS of low-risk women, a broader range of non-cationic proteins were associated with HIV neutralizing activity in HESN and HIV-positive female sex workers. These results indicate that novel protein factors found in CVS of women with high-risk sexual practices may have inherent antiviral activity, or are involved in other aspects of anti-HIV host defense, and warrant further exploration into their mode of action.


Pr2FeCrO6: A Type I Multiferroic.
We synthesized double perovskite Pr2FeCrO6 by solid-state method. Analysis of its X-ray powder diffraction shows that the compound crystallizes in a centrosymmetric structure with space group Pbnm. Our X-ray photoelectron spectroscopy (XPS) studies show that all the cations are present in +3 oxidation state. Magnetization studies of Pr2FeCrO6 show that the material is paramagnetic at room temperature and undergoes a magnetic transition below TCM = 250 K. We observe clear magnetic hysteresis loop, for example, below 150 K. A low remnant magnetization Mr, ∼0.05 μB/f. u., is inferred from the observed magnetic hysteresis loop. 57Fe Mössbauer study at 25 K shows a high hyperfine magnetic field of ∼53 T at the Fe nucleus, which corresponds to a magnetic moment of ∼6-7 μB/Fe. These two results together suggest a ferrimagnetic (nearly compensated or canted) ordering of the Fe moments. Mössbauer studies close to the ferrimagnetic ordering temperature suggest interesting magnetic relaxation effects. A dielectric anomaly observed at TCE = 453 K signals a ferroelectric ↔ paraelectric phase transition. We observe at room temperature a clear and well-defined ferroelectric hysteresis loop, PS = 1.04 μC/cm2, establishing ferroelectricity in the material. From these results, we conclude that Pr2FeCrO6 is a type I multiferroic (TCE > TCM).


Antibodies against citrullinated proteins enhance tissue injury in experimental autoimmune arthritis.
Antibodies against citrullinated proteins are specific and predictive markers for rheumatoid arthritis although the pathologic relevance of these antibodies remains unclear. To investigate the significance of these autoantibodies, collagen-induced arthritis (CIA) in mice was used to establish an animal model of antibody reactivity to citrullinated proteins. DBA/1J mice were immunized with bovine type II collagen (CII) at days 0 and 21, and serum was collected every 7 days for analysis. Antibodies against both CII and cyclic citrullinated peptide, one such citrullinated antigen, appeared early after immunization, before joint swelling was observed. Further, these antibodies demonstrated specific binding to citrullinated filaggrin in rat esophagus by indirect immunofluorescence and citrullinated fibrinogen by Western blot. To evaluate the role of immune responses to citrullinated proteins in CIA, mice were tolerized with a citrulline-containing peptide, followed by antigen challenge with CII. Tolerized mice demonstrated significantly reduced disease severity and incidence compared with controls. We also identified novel murine monoclonal antibodies specific to citrullinated fibrinogen that enhanced arthritis when coadministered with a submaximal dose of anti-CII antibodies and bound targets within the inflamed synovium of mice with CIA. These results demonstrate that antibodies against citrullinated proteins are centrally involved in the pathogenesis of autoimmune arthritis.


Optimizing 802.15.4 Outdoor IoT Sensor Networks for Aerial Data Collection.
Rural IoT sensor networks, prevalent in environmental monitoring and precision agriculture, commonly operate over some variant of the IEEE 802.15.4 standard. Data collection from these networks is often challenging, as they may be deployed in remote regions where existing backhaul infrastructure is expensive or absent. With the commercial and industrial success of Unmanned Aircraft Systems (UAS), there is understandable interest in using UASs for delay tolerant data collection from 802.15.4 IoT sensor networks. In this study, we investigate how to optimize 802.15.4 networks for aerial data collection, which, unlike other wireless standards, has not received rigorous evaluation for three-dimensional aerial communication. We analyze experimental measurements from an outdoor aerial testbed, examining how factors, such as antenna orientation, altitude, antenna placement, and obstruction, affect signal strength and packet reception rate. In our analysis, we model and predict the quality of service for aerial data collection, based on these network configuration variables, and contrast that with the Received Signal Strength Indication (RSSI)-a commonly used signal strength metric. We find that network configuration plays a significant role in network quality, which RSSI, a mediator variable, struggles to account for in the presence of high packet loss. We conclude with a discussion of strategies for optimizing sensor network configuration for aerial data collection, in light of our results.


Multiplex PCR as a tool for validating plasmid content of Borrelia burgdorferi.
Borrelia burgdorferi has an unusual genomic structure containing 21 plasmids. These plasmids carry genes that are essential for infectivity and survival of the spirochetes in vivo. Several plasmids are lost during cultivation in vitro, which might lead to a heterogeneous population after multiple passages and loss of infectivity in laboratory animals. Herein, we present a simple and inexpensive multiplex PCR method that detects the complete plasmid profile of B. burgdorferi B31 in just two PCR tubes.


Fetal hemoglobin in umbilical cord blood in preeclamptic and normotensive pregnancies: A cross-sectional comparative study.
Preeclampsia (PE) is associated with increased fetal hemoglobin (HbF) in the maternal circulation but its source is unknown. To investigate whether excessive HbF is produced in the placenta or the fetus, the concentration of HbF (cHbF) in the arterial and venous umbilical cord blood (UCB) was compared in 15825 normotensive and 444 PE pregnancies. The effect of fetal gender on cHbF was also evaluated in both groups. Arterial and venous UCB sampled immediately after birth at 36-42 weeks of gestation were analyzed for total Hb concentration (ctHb) (g/L) and HbF% using a Radiometer blood gas analyzer. Non-parametric tests were used for statistical comparison and P values < 0.05 were considered significant. Our results indicated higher cHbF in venous compared to arterial UCB in both normotensive (118.90 vs 117.30) and PE (126.75 vs 120.12) groups. In PE compared to normotensive pregnancies, a significant increase was observed in arterial and venous ctHb (171.00 vs 166.00 and 168.00 vs 163.00, respectively) while cHbF was only significantly increased in venous UCB (126.75 vs 118.90). The pattern was similar in both genders. These results indicate a substantial placental contribution to HbF levels in UCB, which increases in PE and is independent of fetal gender, suggesting the elevated cHbF evident in PE results from placental dysfunction.


Clavulanate combinations with mecillinam, cefixime or cefpodoxime against ESBL-producing Enterobacterales frequently associated with blaOXA-1 in a paediatric population with febrile urinary tract infections.
Oral treatment of febrile urinary tract infections (FUTIs) can be impaired by MDR Enterobacterales often combining ESBL and inhibitor-resistant genes. We studied the impact of β-lactamases and Enterobacterales' genotypes on the cefixime, cefpodoxime and mecillinam ± amoxicillin/clavulanate MICs. In this multicentric study, we included 251 previously whole-genome-sequenced ESBL-producing Enterobacterales, isolated in French children with FUTIs. The MICs of cefixime, cefpodoxime, mecillinam alone and combined with amoxicillin/clavulanate were determined and analysed with respect to genomic data. We focused especially on the isolates' ST and their type of β-lactamases. Clinical outcomes of patients who received cefixime + amoxicillin/clavulanate were also analysed. All isolates were cefixime and cefpodoxime resistant. Disparities depending on blaCTX-M variants were observed for cefixime. The addition of amoxicillin/clavulanate restored susceptibility for cefixime and cefpodoxime in 97.2% (MIC50/90 of 0.38/0.75 mg/L) and 55.4% (MIC50/90 of 1/2 mg/L) of isolates, respectively, whatever the ST, the blaCTX-M variants or the association with inhibitor-resistant β-lactamases (34.2%). All isolates were susceptible to mecillinam + amoxicillin/clavulanate with MIC50/90 of 0.19/0.25 mg/L, respectively. Neither therapeutic failure nor any subsequent positive control urine culture were reported for patients who received cefixime + amoxicillin/clavulanate as an oral relay therapy (n = 54). Despite the frequent association of ESBL genes with inhibitor-resistant β-lactamases, the cefixime + amoxicillin/clavulanate MICs remain low. The in vivo efficacy of this combination was satisfying even when first-line treatment was ineffective. Considering the MIC distributions and pharmacokinetic parameters, mecillinam + amoxicillin/clavulanate should also be an alternative to consider when treating FUTIs in children.


Sesquiterpene lactone content and overall quality of fresh-cut witloof chicory (Cichorium intybus L. var. foliosum Hegi) as affected by different washing procedures.
Fresh-cut chicory was produced by four different processing lines testing cold and warm water (4/45 °C, 120 s) and cold water containing cysteine (0.5 mM) against an unwashed control. Levels of sesquiterpene lactones (SLs), sensory, and color attributes as well as O2 and CO2 levels in the modified atmosphere of the fresh-cut produce stored in consumer-sized bags (4 °C, 10 days) were monitored. All washing procedures applied significantly reduced the SL contents and bitterness in fresh-cut chicory. Warm water washing was most effective, reducing the total SL levels by 60.9-64.5%. Sensory and color attributes were also retained best by warm water washing, suggesting such treatments to be suitable measures to improve the quality and reduce the bitterness of fresh-cut chicory. SL profiles changed substantially in all samples during storage, revealing lactucopicrin to be the major SL directly after processing, whereas 11(S),13-dihydrolactuin prevailed at the end of storage.


Sexual Dysfunction in Epilepsy and the Role of Anti-Epileptic Drugs.
Sexual dysfunction is very common in patients with epilepsy. We sought to review the published literature around sexual dysfunction in epilepsy, and particularly the role played by anticonvulsant drugs. We searched all related articles on PubMed using the search terms sexual dysfunction, various AEDs and epilepsy, and restricted the search to English language articles. The most common manifestations of sexual dysfunction in patients with epilepsy are hyposexuality and erectile dysfunction. The cause of this dysfunction is multifactorial and overlapping, and includes changes in the levels of sex hormones, anti-convulsants, the epilepsy itself and psychosocial factors. Traditional anti-convulsants which induce the cytochrome P450 enzyme system have the highest rates of sexual dysfunction, which is primarily mediated by changes in sex hormone levels. Sexual dysfunction associated with newer anti-convulsants is likely to occur through different mechanisms. Some anti-convulsants such as oxcarbazepine and lamotrigine may improve sexual function, but can also rarely be associated with sexual dysfunction. Management of sexual dysfunction thought to be caused by anti-convulsants should include the cessation of the offending drug, and consideration of switching to alternative anti-convulsants that have been reported to improve sexual function such as oxcarbazepine, and lamotrigine.


Non-lethal detection of DNA from Cichlidogyrus spp. (Monogenea, Ancyrocephalinae) in gill mucus of the Nile tilapia Oreochromis niloticus.
Infection of Nile tilapia Oreochromis niloticus by monogeneans of the genus Cichlidogyrus is harmful. Currently, diagnosis of this infection is based on invasive techniques and the identification of isolated parasites by their morphology. To facilitate diagnosis, we have developed a non-lethal polymerase chain reaction (PCR) test for detection of Cichlidogyrus spp. DNA in the gill mucus of O. niloticus, using 5 pairs of specific primers based on Cichlidogyrus sclerosus 28S rRNA (Cicly 1 to Cicly 5) which generate fragments of approximately 188, 180, 150, 159 and 189 bp, respectively. PCR specificity was tested using genomic DNA extracted individually from 175 isolated Cichlidogyrus spp., 75 Gyrodactylus cichlidarum and 75 endopararasitic Enterogyrus spp., as well as from 75 protozoans Trichodina spp. The Cicly primers were used to detect Cichlidogyrus spp. DNA in mucus from the gills of 23 Nile tilapia confirmed to be infected with the parasite. Negative controls consisted of 45 uninfected Nile tilapia. The limit of sensitivity of the assay was 1.2 ng of purified parasite DNA. The Cicly primers did not amplify DNA from the mucus of non-infected Nile tilapia, G. cichlidarum, Trichodina spp. or Enterogyrus spp. In all cases, the sensitivity and specificity of the test were 100%. The sequences of all the amplified fragments showed a high similarity to that of the 28S rRNA region of C. sclerosus (93 to 100% identical to GenBank Accession No. DQ157660.1). We provide evidence for a safe and non-invasive DNA-based diagnostic method for the presence of Cichlidogyrus in the gill mucus of O. niloticus.


Parvocellular and magnocellular contributions to the initial generators of the visual evoked potential: high-density electrical mapping of the "C1" component.
The C1 component of the VEP is considered to index initial afference of retinotopic regions of human visual cortex (V1 and V2). C1 onsets over central parieto-occipital scalp between 45 and 60 ms, peaks between 70 and 100 ms, and then resolves into the following P1 component. By exploiting isoluminant and low-contrast luminance stimuli, we assessed the relative contributions of the Magnocellular (M) and Parvocellular (P) pathways to generation of C1. C1 was maximal at 88 ms in a 100% luminance contrast condition (which stimulates both P and M pathways) and at 115 ms in an isoluminant chromatic condition (which isolates contributions of the P pathway). However, in a 4% luminance contrast condition (which isolates the M pathway), where the stimuli were still clearly perceived, C1 was completely absent. Absence of C1 in this low contrast condition is unlikely to be attributable to lack of stimulus energy since a robust P1-N1 complex was evoked. These data therefore imply that C1 may be primarily parvocellular in origin. The data do not, however, rule out some contribution from the M system at higher contrast levels. Nonetheless, that the amplitude of C1 to P-isolating isoluminant chromatic stimuli is equivalent to that evoked by 100% contrast stimuli suggests that even at high contrast levels, the P system is the largest contributor. These data are related to intracranial recordings in macaque monkeys that have also suggested that the initial current sink in layer IV may not propagate effectively to the scalp surface when M-biased stimuli are used. We also discuss how this finding has implications for a long tradition of attention research that has used C1 as a metric of initial V1 afference in humans. C1 has been repeatedly interrogated for potential selective attentional modulations, particularly in spatial attentional designs, under the premise that modulation of this component, or lack thereof, would be evidence for or against selection at the initial inputs to visual cortex. Given the findings here, we would urge that in interpreting C1 effects, a consideration of the dominant cellular contributions will be necessary. For example, it is plausible that spatial attention mechanisms could operate primarily through the M system and that as such C1 may not always represent an adequate dependent measure in such studies.


The effect of prenatal and postnatal caffeine exposure on pentylentetrazole induced seizures in the non-epileptic and epileptic offsprings.
Caffeine, a central nervous system stimulant, has been reported to modulate seizure activity in various studies. In this study the effects of caffeine exposure on the pentylenetetrazole (PTZ) induced seizure thresholds and seizure stages in the Wistar and genetic absence epilepsy model offsprings were examined. Adult female and male Wistar rats and genetic absence epilepsy rats from Strasbourg (GAERS) consumed caffeine dissolved in water (0.3 g/L) before conception, during the gestational periods and lactation period whereas control groups of each strain received tap water. All offsprings at postnatal day 30 (PN30) subjected to 70 mg/kg of PTZ were evaluated in terms of overall seizure stages, the latency to the first generalized seizure and the c-Fos protein activity in the brain regions of somatosensorial cortex (SSCx), reticular thalamic nucleus (Rt), ventrobasal thalamus (VB), centromedial nucleus (CM) and lateral geniculate nucleus (LGN). The Wistar caffeine group had significantly shorter latency to the first generalized seizure (1.53 ± 0.49 min) comparing to the Wistar control offsprings (3.40 ± 0.68 min). GAERS caffeine group (6.52 ± 2.48 min) showed significantly longer latency comparing to Wistar caffeine group (1.53 ± 0.49 min). Although statistically not significant, GAERS caffeine group showed a longer latency comparing to the GAERS control group (4.71 ± 1.82 min). In all regions of SSCx, Rt, VB, CM and LGN, GAERS caffeine group had lower c-Fos protein expression comparing to the GAERS control group (p < 0.05). Wistar caffeine rats had lower expression of c-Fos protein comparing to the Wistar control group only in SSCx. In CM, GAERS rats expressed lower c-Fos protein comparing to the Wistar control (p < 0.05). In conclusion differential effects of caffeine in the seizure modulation may involve c-Fos protein activity-dependent protection mechanisms.


Cumulative and irreversible cardiac mitochondrial dysfunction induced by doxorubicin.
Interference with mitochondrial calcium regulation is proposed to be a primary causative event in the mechanism of doxorubicin-induced cardiotoxicity. We previously reported disruption of mitochondrial calcium homeostasis after chronic doxorubicin administration (Solen et al. Toxicol. Appl. Pharmacol, 129: 214-222, 1994). The present study was designed to characterize the dose-dependent and cumulative interference with mitochondrial calcium regulation and to assess the reversibility of this functional lesion. Sprague Dawley rats were treated with 2 mg/kg/week doxorubicin s.c. for 4-8 weeks. With succinate as substrate, cardiac mitochondria isolated from rats after 4 weeks of treatment with doxorubicin expressed a lower calcium loading capacity compared with control. This suppression of calcium loading capacity increased with successive doses to 8 weeks of treatment (P < 0.05) and persisted for 5 weeks after the last doxorubicin injection, and was corroborated by dose-dependent and irreversible histopathological changes. Preincubation of mitochondria with tamoxifen, DTT, or monobromobimane did not reverse the diminished calcium loading capacity caused by doxorubicin. In contrast, incubation with cyclosporin A abolished any discernible difference in mitochondrial calcium loading capacity between doxorubicin-treated and saline-treated rats. The decrease in cardiac mitochondrial calcium loading capacity was not attributable to bioenergetic changes in the electron transport chain, because the mitochondrial coupling efficiency was not altered by doxorubicin treatment. However, the ADP/ATP translocase content was significantly lower in mitochondria from rats that received 8 weeks of doxorubicin treatment. These data indicate that doxorubicin treatment in vivo causes a dose-dependent and irreversible decrease in mitochondrial calcium loading capacity. Suppression of adenine nucleotide translocase content may be a key factor altering the calcium-dependent regulation of the mitochondrial permeability transition pore, which may account for the cumulative and irreversible loss of myocardial function in patients receiving doxorubicin chemotherapy.


Sulindac increases the expression of APC mRNA in malignant colonic epithelial cells: an in vitro study.
Sulindac is a non-steroidal anti-inflammatory drug which induces regression of colonic polyps in patients with familial adenomatous polyposis. Animal and in vitro studies have shown that both the sulphide metabolite of sulindac, which is able to inhibit cyclo-oxygenase, and the sulphone metabolite, which lacks this ability, are able to inhibit the growth of colonic carcinoma cells. The exact mechanism by which these effects occurs is not known. To examine the effect of sulindac sulphide and sulindac sulphone on the expression of APC messenger RNA (mRNA), and on the proliferation of colonic carcinoma cells in vitro. The colonic carcinoma cell line LIM 1215 was treated with sulindac sulphide and sulindac sulphone (10 microM or 100 microM) for 24 hours. Total RNA was extracted and APC mRNA was quantitated using competitive reverse transcription polymerase chain reaction. Measurements of cell number, cell proliferation, and prostaglandin E2 concentrations were also made. A significant increase in APC mRNA was observed after treatment with 10 microM of both sulindac sulphide and sulindac sulphone (control: 37.2 (19.7); 10 microM sulindac sulphide: 129 (112.8); 10 microM sulindac sulphone: 207.7 (102.9) pg/(g total RNA) (p < 0.05). Prostaglandin E2 concentrations were significantly reduced after treatment with sulindac sulphide, but not after sulindac sulphone. Both agents produced a dose dependent reduction in cell numbers and cell proliferation, which was more noticeable after treatment with sulindac sulphide. Both sulindac sulphide and sulindac sulphone inhibit the growth of carcinoma cells in vitro and cause an increase in APC mRNA. The effect of these agents on colonic carcinogenesis is not mediated entirely by means of an inhibition of prostaglandin biosynthesis.


Comparison of cell cultures for rapid isolation of enteroviruses.
Cell culture isolation is still the most reliable method for the detection of enteroviruses from clinical specimens. Rapid diagnosis of enterovirus infection affects patient management. To increase yield and enhance the rapidity of enterovirus isolation in cell cultures, we used Buffalo green monkey kidney (BGM) cells and subpassages of primary human embryonic kidney (HEK) cells in addition to the human diploid fibroblast (MRC-5) cells and primary cynomolgus or rhesus monkey kidney (MK) cells routinely used for enterovirus culturing. Growth characteristics of enteroviruses from 421 specimens were studied. All specimens were cultured in MRC-5, MK, and BGM cells, and 204 of these specimens were also cultured in HEK cells. Forty-two percent of the enteroviruses became positive within 3 days, and 85% did so within 7 days. MRC-5 cells provided the highest yield of enteroviruses overall and were the best cell type for the recovery of poliovirus and echovirus. MK cells provided the second best yield but were more useful than MRC-5 cells for coxsackievirus. BGM cells supported the growth of additional isolates of coxsackievirus and enhanced the speed of isolation. HEK cells supported the growth of additional isolates of both coxsackievirus and echovirus, but subculturing was always required for definite enterovirus cytopathic effects. The recovery rate increased 11% when two additional cell lines were used. The use of two tubes of MK cells significantly increased the yield of all enterovirus types. We conclude that the use of multiple appropriate cell lines increases yield and enhances the rapidity of enterovirus isolation.


CXCL1 but not IL-6 is required for recurrent herpetic stromal keratitis.
Herpetic stromal keratitis (HSK) is characterized by an inflammatory response that includes neutrophils, macrophages, NK cells, and T cells. The factors that are responsible for this inflammation are proinflammatory cytokines and chemokines. Many of these factors have been defined for primary disease, but relatively few have been investigated during recurrent HSK. The present study was designed to determine the role that two of these factors, IL-6 and CXCL1, play during recurrent HSK. Results clearly indicate that unlike primary disease, IL-6 plays no role in recurrent HSK. However, the presence of CXCL1 is required for recurrent HSK as evidenced by the lack of corneal disease in mice treated with anti-CXCL1 Ab. This was confirmed using mice lacking the primary receptor for CXCL1, CXCR2. Corneal disease in this strain was significantly reduced compared with wild-type C57BL/6 controls. Unexpectedly, lack of disease occurs even though CXCL1 knockout mice display increased viral shedding at the cornea. The primary mechanism that CXCL1 plays during disease is its ability to stimulate neutrophils to infiltrate the cornea following reactivation. This paper provides further evidence that primary HSK and recurrent HSK possess overlapping yet distinct disease mechanisms.


The risk functions incorporated in Riscard 2002: a software for the prediction of cardiovascular risk in the general population based on Italian data.
The purpose of this analysis was to produce risk functions for the prediction of cardiovascular diseases based on Italian epidemiological data and suitable for the use in a PC program dedicated to the estimate of risk. Three studies were used for the purpose: the Italian Rural Areas of the Seven Countries Study, the Gubbio Population Study and the ECCIS study, for a total of 9771 men and women aged 35 to 74 years and followed for a period lasting 5 to 6 years. The risk factors used for the prediction of cardiovascular events were sex, age, body mass index (derived from height and weight), mean blood pressure (derived from systolic and diastolic blood pressures), non-HDL cholesterol (derived from total and HDL cholesterol), HDL cholesterol, diabetes (yes-no), heart rate, and daily cigarette consumption. The endpoints were the first major coronary event, the first major cerebrovascular event, and the first major cardiovascular event (either one between the previous two plus major peripheral artery diseases). The model employed for the analysis was the accelerated failure time model. Having excluded those already presenting with a cardiovascular disease and those with missing values, a total of 9089 subjects were included in the models. In a period lasting 5 or 6 years, a total of 211 coronary, 64 cerebrovascular and 269 cardiovascular events occurred and were considered for analysis. Coefficients from the coronary model suggested a significant association of all risk factors except body mass index and diabetes (marginal significance). Coefficients from the cerebrovascular model suggested a significant association limited to age and mean blood pressure. Coefficients from the cardiovascular model suggested a significant association of all risk factors except body mass index. The discrimination between cases and non-cases was satisfactory with proportions of 37.0, 52.3 and 37.8% of observed cases in decile 10 of the distribution of the estimated risk for the three endpoints respectively. The three models were used as a mathematical core for the construction of a PC software for the prediction of major cardiovascular events in Italy.


Unexpected patterns of fisheries collapse in the world's oceans.
Understanding which species are most vulnerable to human impacts is a prerequisite for designing effective conservation strategies. Surveys of terrestrial species have suggested that large-bodied species and top predators are the most at risk, and it is commonly assumed that such patterns also apply in the ocean. However, there has been no global test of this hypothesis in the sea. We analyzed two fisheries datasets (stock assessments and landings) to determine the life-history traits of species that have suffered dramatic population collapses. Contrary to expectations, our data suggest that up to twice as many fisheries for small, low trophic-level species have collapsed compared with those for large predators. These patterns contrast with those on land, suggesting fundamental differences in the ways that industrial fisheries and land conversion affect natural communities. Even temporary collapses of small, low trophic-level fishes can have ecosystem-wide impacts by reducing food supply to larger fish, seabirds, and marine mammals.


Comparison of patients with different pathologies in terms of shoulder protraction and scapular asymmetry.
[Purpose] The aim of this study was to compare cases with different shoulder and cervical pathologies in terms of shoulder protraction and scapular asymmetry. [Methods] A total of 216 patients, aged between 30-70 years, were included, 108 of which were in the patient group (subacromial impingement, rotator cuff problems, adhesive capsulitis, disc herniations) and 108 of which were in the control group. The control group consisted of cases with no prior neck and shoulder problems or pain. Pain was evaluated using the visual analogue scale (VAS); the asymmetry of scapula was evaluated using the Lateral Scapular Slide Test (LSST) with two additional positions; and the protraction of the scapula was evaluated using the shoulder protraction test. [Results] According to the data obtained, the affected side scapular asymmetry and protraction in the patient group were significantly greater than in the control group. When the patient groups were compared in terms of different pathologies, there were no differences between scapular asymmetry and shoulder protraction. [Conclusion] In conclusion, the pathologies of the neck and shoulder were found to cause scapular asymmetry and shoulder protraction. However, patients with different pathologies had similar scapular asymmetry and shoulder protraction.


Human sarcopenia reveals an increase in SOCS-3 and myostatin and a reduced efficiency of Akt phosphorylation.
Age-related skeletal muscle sarcopenia is linked with increases in falls, fractures, and death and therefore has important socioeconomic consequences. The molecular mechanisms controlling age-related muscle loss in humans are not well understood, but are likely to involve multiple signaling pathways. This study investigated the regulation of several genes and proteins involved in the activation of key signaling pathways promoting muscle hypertrophy, including GH/STAT5, IGF-1/Akt/GSK-3beta/4E-BP1, and muscle atrophy, including TNFalpha/SOCS-3 and Akt/FKHR/atrogene, in muscle biopsies from 13 young (20 +/- 0.2 years) and 16 older (70 +/- 0.3 years) males. In the older males compared to the young subjects, muscle fiber cross-sectional area was reduced by 40-45% in the type II muscle fibers. TNFalpha and SOCS-3 were increased by 2.8 and 1.5 fold, respectively. Growth hormone receptor protein (GHR) and IGF-1 mRNA were decreased by 45%. Total Akt, but not phosphorylated Akt, was increased by 2.5 fold, which corresponded to a 30% reduction in the efficiency of Akt phosphorylation in the older subjects. Phosphorylated and total GSK-3beta were increased by 1.5 and 1.8 fold, respectively, while 4E-BP1 levels were not changed. Nuclear FKHR and FKHRL1 were decreased by 73 and 50%, respectively, with no changes in their atrophy target genes, atrogin-1 and MuRF1. Myostatin mRNA and protein levels were significantly elevated by 2 and 1.4 fold. Human sarcopenia may be linked to a reduction in the activity or sensitivity of anabolic signaling proteins such as GHR, IGF-1, and Akt. TNFalpha, SOCS-3, and myostatin are potential candidates influencing this anabolic perturbation.


Chronic treatment with caffeine and its withdrawal modify the antidepressant-like activity of selective serotonin reuptake inhibitors in the forced swim and tail suspension tests in mice. Effects on Comt, Slc6a15 and Adora1 gene expression.
Recent preclinical and clinical data suggest that low dose of caffeine enhances the effects of common antidepressants. Here we investigated the effects of chronic administration of caffeine (5mg/kg, twice daily for 14days) and its withdrawal on day 15th on the activity of per se ineffective doses of fluoxetine (5mg/kg) and escitalopram (2mg/kg) given on day 15th. We found decreased immobility time in the forced swim and tail suspension tests in mice in which caffeine was administered simultaneously with antidepressants on day 15th following a 14-day caffeine treatment and no alterations in the spontaneous locomotor activity. A decrease in the level of escitalopram and an increase in the level of caffeine in serum were observed after concomitant administration of these compounds, while the joint administration of caffeine and fluoxetine was not associated with changes in their levels in serum or brain. Caffeine withdrawal caused a decrease in Adora1 mRNA level in the cerebral cortex (Cx). Administration of escitalopram or fluoxetine followed by caffeine withdrawal caused an increase in this gene expression, whereas administration of escitalopram, but not fluoxetine, on day 15th together with caffeine caused a decrease in Adora1 mRNA level in the Cx. Furthermore, antidepressant-like activity observed after joint administration of the tested drugs with caffeine was associated with decreased Slc6a15 mRNA level in the Cx. The results show that withdrawal of caffeine after its chronic intake may change activity of antidepressants with concomitant alterations within monoamine, adenosine and glutamate systems.