Immersion exposure of rainbow trout (Oncorhynchus mykiss) fry to wildtype Flavobacterium psychrophilum induces no mortality, but protects against later intraperitoneal challenge.
Flavobacterium psychrophilum, the causative agent of RTFS or rainbow trout fry syndrome, causes high mortality among hatchery reared rainbow trout (Oncorhynchus mykiss) fry in Europe and the USA. Despite several attempts, no efficient vaccines have yet been developed, the main obstacle being that the fry have to be vaccinated very early, i.e. around 0.2-0.5 g, where RTFS usually starts to give problems in the fish farms. Consequently, only oral or bath vaccines are relevant. Immersion of fry in inactivated or attenuated bacteria has resulted in RPS values of less than 50%. However, the results are biased by the fact that the fish have been challenged by intraperitoneal (ip) or subcutaneous (sc) injection against which an immersion/oral vaccine may not protect. Therefore, the present study was undertaken in order to investigate whether the presumably most potent immersion immunization, i.e. bathing in high titres of non-attenuated isolates of F. psychrophilum, was able to induce immunity to a subsequent ip challenge. Immersion in live bacteria for 30 or 50 min caused no mortality and protected a major fraction of the fry against challenges 26 and 47 days later with RPS values of 88.2 and 60.3%, respectively. Increased specific antibody titres suggested that adaptive immune mechanisms were involved in the protection.


Fractionation of the more active extracts of Geranium molle L.: a relationship between their phenolic profile and biological activity.
Geranium molle L., commonly known as Dove's-foot Crane's-bill or Dovesfoot Geranium, is an herbaceous plant belonging to the Geraniaceae family. Contrary to many other Geranium species, the bioactivity and the phytochemical composition of G. molle seem not to have attracted attention until a recent study from our group regarding the bioactivity of several aqueous and organic extracts of the plant. In particular, we assessed the cytotoxic activity of these extracts against several human tumor cell lines (breast, lung, cervical and hepatocellular carcinomas) and a non-tumor porcine liver primary cell line, inspired by an ethnopharmacological report describing the traditional use of this medicinal plant in some regions of Northeast Portugal for the treatment of cancer. Following this preliminary evaluation, the most active extracts (acetone and methanol) were fractionated by column chromatography and the resulting fractions were evaluated for their antioxidant activity and cytotoxicity against the same cell lines. The bio-guided fractionation of the extracts resulted in several fractions exhibiting improved bioactivity in comparison with the corresponding crude extracts. The fractions obtained from the acetone extract consistently displayed the lowest EC50 and GI50 values and presented the highest content of total phenolic compounds. The phytochemical composition of the most bioactive fractions of the acetone and methanol extracts was also determined and about thirty compounds, mainly flavonoids and phenolic acids, could be identified for the first time in G. molle.


Blood pressure lowering efficacy of beta-blockers as second-line therapy for primary hypertension.
Beta-blockers are one of the more commonly prescribed classes of anti-hypertensive drugs, both as first-line and second-line. To quantify the effect on systolic blood pressure (SBP), diastolic blood pressure (DBP), heart rate and withdrawals due to adverse effects of beta-blocker therapy when given as a second-line drug in adult patients with primary hypertension. CENTRAL (The Cochrane Library 2009, Issue 2), MEDLINE (1966-Aug 2009), EMBASE (1988-Aug 2009) and bibliographic citations of articles and reviews were searched. Double-blind, randomized controlled trials comparing a beta-blocker in combination with a drug from another class of anti-hypertensive drugs compared with that drug alone for a duration of 3 to 12 weeks in patients with primary hypertension were included. Two reviewers independently extracted the data and assessed trial quality of each included study. 20 double-blind RCTs evaluated the BP lowering efficacy of beta-blockers as second-line drug in 3744 hypertensive patients (baseline BP of 158/102 mmHg; mean duration of 7 weeks). The BP reduction from adding a beta-blocker as the second drug was estimated by comparing the difference in BP reduction between the combination and monotherapy groups. A reduction in BP was seen with adding a beta-blocker to thiazide diuretics or calcium channel blockers at doses as low as 0.25 times the manufacturer's recommended starting dose. The BP lowering efficacy of beta-blockers as a second drug was 6/4 mmHg at 1 times the starting dose and 8/6 mmHg at 2 times the starting dose. Beta-blockers reduced heart rate by 10 beats/min at 1 to 2 times the starting dose. Beta-blockers did not statistically significantly increase withdrawals due to adverse effects but this was likely due to the lack of reporting of this outcome in 35% of the included RCTs. Addition of a beta-blocker to diuretics or calcium-channel blockers reduces BP by 6/4mmHg at 1 times the starting dose and by 8/6 mmHg at 2 times the starting dose. When the blood pressure lowering effect of beta-blockers from this review was compared to that of thiazide diuretics from our previous review (Chen 2009), second-line beta-blockers reduce systolic BP to the same extent as second-line thiazide diuretics, but reduce diastolic BP to a greater degree. The different effect on diastolic BP means that beta-blockers have little or no effect on pulse pressure whereas thiazides cause a significant dose-related decrease in pulse pressure. This difference in the pattern of BP lowering with beta-blockers as compared to thiazides might be the explanation for the fact that beta-blockers appear to be less effective at reducing adverse cardiovascular outcomes than thiazide diuretics, particularly in older individuals.


European citizens' use of E-health services: a study of seven countries.
European citizens are increasingly being offered Internet health services. This study investigated patterns of health-related Internet use, its consequences, and citizens' expectations about their doctors' provision of e-health services. Representative samples were obtained from the general populations in Norway, Denmark, Germany, Greece, Poland, Portugal and Latvia. The total sample consisted of 7934 respondents. Interviews were conducted by telephone. 44 % of the total sample, 71 % of the Internet users, had used the Internet for health purposes. Factors that positively affected the use of Internet for health purposes were youth, higher education, white-collar or no paid job, visits to the GP during the past year, long-term illness or disabilities, and a subjective assessment of one's own health as good. Women were the most active health users among those who were online. One in four of the respondents used the Internet to prepare for or follow up doctors' appointments. Feeling reassured after using the Internet for health purposes was twice as common as experiencing anxieties. When choosing a new doctor, more than a third of the sample rated the provision of e-health services as important. The users of Internet health services differ from the general population when it comes to health and demographic variables. The most common way to use the Internet in health matters is to read information, second comes using the net to decide whether to see a doctor and to prepare for and follow up on doctors' appointments. Hence, health-related use of the Internet does affect patients' use of other health services, but it would appear to supplement rather than to replace other health services.


Neighbourhood social capital: measurement issues and associations with health outcomes.
We compared ecometric neighbourhood scores of social capital (contextual variation) to mean neighbourhood scores (individual and contextual variation), using several health-related outcomes (i.e. self-rated health, weight status and obesity-related behaviours). Data were analysed from 5,900 participants in the European SPOTLIGHT survey. Factor analysis of the 13-item social capital scale revealed two social capital constructs: social networks and social cohesion. The associations of ecometric and mean neighbourhood-level scores of these constructs with self-rated health, weight status and obesity-related behaviours were analysed using multilevel regression analyses, adjusted for key covariates. Analyses using ecometric and mean neighbourhood scores, but not mean neighbourhood scores adjusted for individual scores, yielded similar regression coefficients. Higher levels of social network and social cohesion were not only associated with better self-rated health, lower odds of obesity and higher fruit consumption, but also with prolonged sitting and less transport-related physical activity. Only associations with transport-related physical activity and sedentary behaviours were associated with mean neighbourhood scores adjusted for individual scores. As analyses using ecometric scores generated the same results as using mean neighbourhood scores, but different results when using mean neighbourhood scores adjusted for individual scores, this suggests that the theoretical advantage of the ecometric approach (i.e. teasing out individual and contextual variation) may not be achieved in practice. The different operationalisations of social network and social cohesion were associated with several health outcomes, but the constructs that appeared to represent the contextual variation best were only associated with two of the outcomes.


How children die: classifying child deaths.
To validate a descriptive tool for the causes of child death, which was designed to circumvent problems posed by the analysis of a confidential enquiry. 3 participants from different healthcare backgrounds used clinical data, including the entries on the medical certificate of the cause of death, to classify the root cause of 783 deaths from the Confidential Enquiry into Maternal and Child Health child death review. A bespoke hierarchical system was used. Unanimity of allocation within categories and inter-rater and intra-rater agreement were assessed. Two methods for treating disagreements were compared by assessing their effect upon the apparent incidence of different causes of death. The participants were most consistent in grouping deaths due to trauma, malignancy and sudden infant death. Each was highly consistent in allocating cases to groups (κ 0.85-0.99), but the agreement between participants, although "good", was worse (κ 0.66-0.78). The greatest number of discrepancies was between diseases identified as congenital by the doctor and as chronic medical conditions by others. The method for treating disagreement between participants does not affect the commonest cause of death (trauma) but alters the ranking of the subordinate causes. Agreement within diagnostic categories might be improved by greater training of assessors in the use of the technique. This level of performance compares well with that of other coding systems upon their target groups.


Treg-mediated immunosuppression involves activation of the Notch-HES1 axis by membrane-bound TGF-beta.
Studies in humans and mice show an important role for Tregs in the control of immunological disorders. The mechanisms underlying the immunosuppressive functions of Tregs are not well understood. Here, we show that CD4+ T cells expressing Foxp3 and membrane-bound TGF-beta (TGF-beta(m+)Foxp3+), previously shown to be immunosuppressive in both allergic and autoimmune diseases, activate the Notch1-hairy and enhancer of split 1 (Notch1-HES1) axis in target cells. Soluble TGF-beta and cells secreting similar levels of soluble TGF-beta were unable to trigger Notch1 activation. Inhibition of Notch1 activation in vivo reversed the immunosuppressive functions of TGF-beta(m+)Foxp3+ cells, resulting in severe allergic airway inflammation. Integration of the TGF-beta and Notch1 pathways may be an important mechanism for the maintenance of immune homeostasis in the periphery.


Effects of Anti-Smoking Public Service Announcements on the Attitudes of Korean College Students toward Smoking.
This study aimed to identify the effects of anti-smoking public service announcements on the attitudes of Korean college students toward smoking. This study involved students via convenience sampling from seven universities who were randomly assigned to four groups. All groups completed a preliminary questionnaire, before being shown a public service announcement twice, and then completed a post viewing questionnaire. For announcements with positive messages, the proportion of changes in beliefs and attitudes were 39.1% and 19.8%, respectively, whereas those with negative messages showed a greater proportion of changes in the beliefs (59.7%) and attitudes (40.3%). After adjusting for sex and change in belief, the message types and smoking status were identified as factors affecting the change in the participants attitudes. A negative message resulted in a greater change in attitudes (odds ratio [OR], 3.047; 95% confidence interval [CI], 1.847-5.053). Ever-smokers including current smokers showed a greater positive change in attitude than never-smokers (OR, 6.965; 95% CI, 4.107-11.812). This study found that positive anti-smoking public service announcements were more effective on attitude change than negative messages. Additionally these announcements were more effective among viewers who were current smokers or had a prior smoking experience.


Vector competence of biting midges and mosquitoes for Shuni virus.
Shuni virus (SHUV) is an orthobunyavirus that belongs to the Simbu serogroup. SHUV was isolated from diverse species of domesticated animals and wildlife, and is associated with neurological disease, abortions, and congenital malformations. Recently, SHUV caused outbreaks among ruminants in Israel, representing the first incursions outside the African continent. The isolation of SHUV from a febrile child in Nigeria and seroprevalence among veterinarians in South Africa suggests that the virus may have zoonotic potential as well. The high pathogenicity, extremely broad tropism, potential transmission via both biting midges and mosquitoes, and zoonotic features warrants prioritization of SHUV for further research. Additional knowledge is essential to accurately determine the risk for animal and human health, and to assess the risk of future epizootics and epidemics. To gain first insights into the potential involvement of arthropod vectors in SHUV transmission, we have investigated the ability of SHUV to infect and disseminate in laboratory-reared biting midges and mosquitoes. Culicoides nubeculosus, C. sonorensis, Culex pipiens pipiens, and Aedes aegypti were orally exposed to SHUV by providing an infectious blood meal. Biting midges showed high infection rates of approximately 40-60%, whereas infection rates of mosquitoes were lower than 2%. SHUV successfully disseminated in both species of biting midges, but no evidence of transmission in orally exposed mosquitoes was found. The results of this study show that different species of Culicoides biting midges are susceptible to infection and dissemination of SHUV, whereas the two mosquito species tested were found not to be susceptible.


Polymorphisms of two fucosyltransferase genes (Lewis and Secretor genes) involving type I Lewis antigens are associated with the presence of anti-Helicobacter pylori IgG antibody.
Helicobacter pylori attach to the gastric mucosa with adhesin, which binds to Lewis b (Le(b)) or H type I carbohydrate structures. The Secretor (Se) gene and Lewis (Le) gene are involved in type I Le antigen synthesis. The present study was performed to investigate the possibility that Se and Le gene polymorphisms alter the risk of H. pylori infection. Two hundred thirty-nine participants were genotyped for Se and Le and tested for the presence of anti-H. pylori IgG antibodies. Using the normal gastric mucosa from 60 gastric cancer patients, we assessed immunohistochemically whether type I Le antigen expression depended on the Se and Le genotypes. The H. pylori infection rate was positively associated with the number of Se alleles (se/se group, 45.1%; Se/se group, 64.6%; and Se/Se group, 73.3%) and negatively associated with the number of Le alleles (le/le group, 76.4%; Le/le group, 68.3%; and Le/Le group, 55.6%). When the subjects were classified into three groups [low risk, (se/se, Le/Le) genotype; high risk, (Se/Se, le/le), (Se/Se, Le/le), and (Se/se, le/le) genotypes; moderate risk, other than low- or high-risk group], the odds ratio relative to the low-risk group was 3.30 (95% confidence interval, 1.40-7.78) for the moderate-risk group and 10.33 (95% confidence interval, 3.16-33.8) for the high-risk group. Immunohistochemical analysis supported the finding that Se and Le genotypes affected the expression of H. pylori adhesin ligands. We conclude that Se and Le genotypes affect susceptibility to H. pylori infection.


Uncovering signal transduction networks from high-throughput data by integer linear programming.
Signal transduction is an important process that transmits signals from the outside of a cell to the inside to mediate sophisticated biological responses. Effective computational models to unravel such a process by taking advantage of high-throughput genomic and proteomic data are needed to understand the essential mechanisms underlying the signaling pathways. In this article, we propose a novel method for uncovering signal transduction networks (STNs) by integrating protein interaction with gene expression data. Specifically, we formulate STN identification problem as an integer linear programming (ILP) model, which can be actually solved by a relaxed linear programming algorithm and is flexible for handling various prior information without any restriction on the network structures. The numerical results on yeast MAPK signaling pathways demonstrate that the proposed ILP model is able to uncover STNs or pathways in an efficient and accurate manner. In particular, the prediction results are found to be in high agreement with current biological knowledge and available information in literature. In addition, the proposed model is simple to be interpreted and easy to be implemented even for a large-scale system.


Pathological characteristics of the large renal mass: potential implication for clinical role of renal biopsy.
To assess whether patients with a large renal mass, treated by radical nephrectomy (RN), could have benefited from preoperative renal mass biopsy (RMB). The decision to perform partial nephrectomy (PN) for an organ-confined > 4 cm renal mass can be complex. Albeit often feasible, oncologic safety of PN in this cohort is debated. Yet, a significant portion of large renal masses that undergo RN prove benign or indolent, indicating a potential role for RMB to guide nephron preservation. We queried prospectively maintained databases from three institutions to identify patients who underwent RN for localized > 4 cm renal mass. We excluded patients with nodal or distant metastases. Multivariable analysis assessed how clinicopathologic variables, mass anatomic complexity, and patient comorbidities related to the likelihood of harboring an indolent neoplasm. A total of 702 patients underwent RN for localized > 4 cm renal mass (median tumor size 7.0 cm (IQR 5.5-9.2); 12.8% (n = 90) of patients were diagnosed with oncocytoma/oncocytic neoplasm (n = 27, 3.8%) or chromophobe RCC (n = 63, 9.0%). When stratified by tumor size, indolent tumors comprised 10.1% of 4-7 cm masses, 15.6% of ≥ 7-10 cm masses, and 17.3% of ≥ 10 cm tumors. Upon multivariate analysis, younger age was associated with indolent tumors (p = 0.04, OR 0.97, 95% CI 0.94-0.99). Approximately 1 in 8 patients with a renal mass > 4 cm harbored benign or low risk indolent potential lesions and were associated with younger age. As such, patients with large renal masses for whom risk trade-offs between PN and RN are unclear, present a unique opportunity for greater utilization of RMB.


Allorecognition of purified major histocompatibility complex glycoproteins by cytotoxic T lymphocytes.
To investigate how T cells recognize allogeneic class I proteins encoded by the major histocompatibility complex (MHC), we examined the human cytotoxic T lymphocytes (CTL) elicited in a mixed lymphocyte reaction against a lymphoblastoid B-cell line (JY) whose MHC-class I proteins are HLA-A2 and -B7. By panning the responding T cells on plates that were coated with purified HLA-A2, an essentially pure population of CD8+ anti-HLA-A2 CTL was isolated in a single step and established as a cell line designated A2p. In addition to lysing HLA-A2+ target cells, the A2p cells lysed HLA-A2- cells, including mouse cells (P815), when purified native HLA-A2 was attached to them, but not when denatured HLA-A2 was attached. Thus, contrary to the general rule that T cells recognize sequential antigenic determinants in denatured protein antigens, the alloreactive CTLs appear to recognize determinants that depend upon the native configuration of HLA-A2; however, the possibility that these T cells recognize a peptide adduct persistently associated with purified, soluble HLA-A2 has not been ruled out.


Removal of pedestals and directional ambiguity of optical anemometer signals.
Laser Doppler anemometry permits, in principle, the measurement of both magnitude and direction of components of a particle's velocity vector. Most exiting anemometers, however, permit measurements only with a directional ambiguity of 180 degrees , resulting in errors in certain flow fields. Available methods of eliminating the directional ambiguity of Laser Doppler anemometers are reviewed, covering frequency shifting of the incident and scattered light beams, the use of beams with different polarization properties, and employment of multicolor laser beams. The advantages and disadvantages of existing methods are summarized, and suggestions for alterations are made. Different techniques used to remove the pedestal of laser Doppler anemometer signals are also reviewed. Optical techniques should be employed in any advanced optical anemometer system to avoid dynamic range limitations by electronic bandpass filters. Suggestions are made for advanced optical anemometers employing multielement avalanche photodiodes that can be used for simultaneous measurements of two velocity components. These anemometers incorporate devices to sense the direction of the velocity components and to eliminate optically the pedestal of laser Doppler signals.


Do weather changes influence physical activity level among older adults? - The Generation 100 study.
Understanding how individual and environmental factors impact physical activity (PA) level is important when building strategies to improve PA of older adults. No studies have examined how hour-to-hour weather changes influence PA in older adults or how the association between weather and PA eventually is related to cardiorespiratory fitness (CRF) measured as peak oxygen uptake. The aim of this study was therefore to examine how hour-to-hour changes in weather effects hour-to-hour PA in a cohort of Norwegian older adults across CRF levels, gender and seasons. PA was assessed objectively in 1219 older adults (70-77 years, 51% females) using the Actigraph GT3X+ accelerometer, and quantified as counts·min-1 (CPM). Weather (Norwegian meteorological Institute) and CRF (MetaMax II) were measured objectively. Panel data analysis added a longitudinal dimension when 110.888 hours of weather- and PA data were analyzed. Older adults had a higher PA level in warmer (597 CPM) than colder months (556 CPM) (p<0.01). Fixed effects regression-models revealed that increasing temperatures (per hour) influenced PA positively in both colder and warmer months (all, p<0.01), with greater influence in fitter vs. less fit participants (p<0.01). In warmer months, increasing precipitation negatively influenced PA in both unfit females and unfit males (p<0.01). In colder months, increasing precipitation positively influenced PA for moderately fit to fit males (p<0.01), but not for females and unfit males. To the best of our knowledge, this is the first study to examine the association between weather conditions and objectively-measured-PA among Norwegian older adults. Our findings demonstrates that unfit older adults will be less likely to participate in PA when the weather is unpleasant, compared to those highly fit. The data suggests that the impact of weather should not be ignored when planning public health strategies for increasing PA among older adults.


Effects of nilotinib on leukaemia cells using vibrational microspectroscopy and cell cloning.
Over the last few years, both synchrotron-based FTIR (S-FTIR) and Raman microspectroscopies have helped to better understand the effects of drugs on cancer cells. However, cancer is a mixture of cells with different sensitivity/resistance to drugs. Furthermore, the effects of drugs on cells produce both chemical and morphological changes, the latter could affect the spectra of cells incubated with drugs. Here, we successfully cloned sensitive and resistant leukaemia cells to nilotinib, a drug used in the management of leukaemia. This allowed both the study of a more uniform population and the study of sensitive and resistant cells prior to the addition of the drug with both S-FTIR and Raman microspectroscopies. The incubation with nilotinib produced changes in the S-FTIR and Raman spectra of both sensitive and resistant clones to nilotinib. Principal component analysis was able to distinguish between cells incubated in the absence or presence of the drug, even in the case of resistant clones. The latter would confirm that the spectral differences between the so-called resistant clonal cells prior to and after adding a drug might reside on those more or less sensitive cells that have been able to remain alive when they were collected to be studied with S-FTIR or Raman microspectroscopies. The data presented here indicate that the methodology of cell cloning can be applied to different types of malignant cells. This should facilitate the identification of spectral biomarkers of sensitivity/resistance to drugs. The next step would be a better assessment of sensitivity/resistance of leukaemia cells from patients which could guide clinicians to better tailor treatments to each individual patient.


AICD nuclear signaling and its possible contribution to Alzheimer's disease.
Altered proteolytic processing of the β-amyloid precursor protein (APP) is a central event in familial and sporadic Alzheimer's disease (AD). In a process termed regulated intramembrane proteolysis (RIP), APP first undergoes ectodomain shedding executed either by α- secretases at the plasma membrane or by β-secretase in the endosomal compartment. The remaining membrane-anchored stubs are cleaved within the membrane plane by the γ-secretase complex, releasing the APP intracellular domain (AICD) into the cytosol and leading to the generation of the Aβ peptide in the amyloidogenic pathway that is initiated by β-secretase. The Aβ peptides aggregate to form soluble oligomers and finally deposit into amyloid plaques that are a hallmark of AD. Recent evidence indicates a role for Aβ oligomers in regulating synaptic plasticity with excess amounts of oligomers disrupting synaptic function. The amyloid cascade hypothesis of AD is centered on the Aβ peptide, the APP fragment that has been most intensely studied, while other cleavage products have been largely neglected. The secreted ectodomain generated after α-cleavage in the non-amyloidogenic pathway has neurotrophic and neuroproliferative activities, thus opposing the neurotoxicity observed with high concentrations of Aβ. Further, in analogy to many other membrane proteins that are subject to RIP, AICD can translocate to the nucleus to regulate transcription. Many RIP substrates are localized to the synapse and thus could convey a direct signal from the synapse to the nucleus upon cleavage. Evidence indicates that only the amyloidogenic pathway generates AICD capable of nuclear signaling, due to the subcellular compartmentalization of APP processing. In aging and sporadic AD there is an increase in β-secretase levels and activity generating more Aβ peptides and concomitantly leading to an increase in AICD nuclear signaling. In this review, I summarize the current knowledge on AICD nuclear signaling and propose mechanisms to explain how this physiological function of APP might impact the pathology seen in AD.


What role does multisensory integration play in the visuotactile perception of texture?
Previous research has shown that vision and touch are both effective at many roughness discrimination tasks; however, there is no evidence that using both senses simultaneously improves discrimination performance. We investigated the nature of this failure to integrate multisensory inputs, using three varieties of forced-choice discrimination tasks. In Experiment 1, visual, tactile and bimodal roughness discriminations were made between pairs of fabric stimuli. Bimodal discriminations were typically performed with a sensitivity somewhere between that observed for the unimodal presentations. In Experiment 2, a similar design was used except that during the stimulus presentation, one interval contained a unimodal (vision or touch) stimulus, the other interval a bimodal stimulus presentation. Bias toward the bimodal interval would indicate an increase in the magnitude of perceived roughness for such presentations. No such bias was found. In Experiment 3, participants made single-interval, bimodal discriminations, determining whether a rough stimulus was presented to touch, to vision, to both modalities, or to neither modality. The improved performance seen for the dual-target vs. single-target presentations was best modelled as arising from a trialwise division of attention between vision and touch. Overall, these results suggest that vision and touch act as independent sources of roughness information, where the necessity to divide attention across both modalities reduces the discriminative ability in (or information available from) each of these individual modalities.


Efficient dengue virus (DENV) infection of human muscle satellite cells upregulates type I interferon response genes and differentially modulates MHC I expression on bystander and DENV-infected cells.
Dengue virus (DENV) is a mosquito-borne flavivirus that causes an acute febrile disease in humans, characterized by musculoskeletal pain, headache, rash and leukopenia. The cause of myalgia during DENV infection is still unknown. To determine whether DENV can infect primary muscle cells, human muscle satellite cells were exposed to DENV in vitro. The results demonstrated for the first time high-efficiency infection and replication of DENV in human primary muscle satellite cells. Changes in global gene expression were also examined in these cells following DENV infection using Affymetrix GeneChip analysis. The differentially regulated genes belonged to two main functional categories: cell growth and development, and antiviral type I interferon (IFN) response genes. Increased expression of the type I IFN response genes for tumour necrosis factor-related apoptosis-inducing ligand (TRAIL), melanoma-derived antigen 5 (MDA-5), IFN-gamma-inducible protein 10 (IP-10), galectin 3 soluble binding protein (LGals3BP) and IFN response factor 7 (IRF7) was confirmed by quantitative RT-PCR. Furthermore, higher levels of cell-surface-bound intracellular adhesion molecule-1 (ICAM-1) and soluble ICAM-1 in the cell-culture medium were detected following DENV infection. However, DENV infection impaired the ability of the infected cells in the culture medium to upregulate cell-surface expression of MHC I molecules, suggesting a possible mechanism of immune evasion by DENV. The findings of this study warrant further clinical research to identify whether muscle cells are targets for DENV infection during the acute stage of the disease in vivo.


Parsing a cognitive task: a characterization of the mind's bottleneck.
Parsing a mental operation into components, characterizing the parallel or serial nature of this flow, and understanding what each process ultimately contributes to response time are fundamental questions in cognitive neuroscience. Here we show how a simple theoretical model leads to an extended set of predictions concerning the distribution of response time and its alteration by simultaneous performance of another task. The model provides a synthesis of psychological refractory period and random-walk models of response time. It merely assumes that a task consists of three consecutive stages-perception, decision based on noisy integration of evidence, and response-and that the perceptual and motor stages can operate simultaneously with stages of another task, while the central decision process constitutes a bottleneck. We designed a number-comparison task that provided a thorough test of the model by allowing independent variations in number notation, numerical distance, response complexity, and temporal asynchrony relative to an interfering probe task of tone discrimination. The results revealed a parsing of the comparison task in which each variable affects only one stage. Numerical distance affects the integration process, which is the only step that cannot proceed in parallel and has a major contribution to response time variability. The other stages, mapping the numeral to an internal quantity and executing the motor response, can be carried out in parallel with another task. Changing the duration of these processes has no significant effect on the variance.


The extended retrosigmoid approach: an alternative to radical cranial base approaches for posterior fossa lesions.
The extended retrosigmoid approach is presented as a simple and safe modification of the traditional retrosigmoid approach, with increased exposure resulting from a limited mastoidectomy and skeletonization of the sigmoid sinus. Patients with posterior fossa vascular lesions treated with the extended retrosigmoid approach between 1997 and 2003 were reviewed. A detailed description of the surgical approach, as well as case illustrations, is provided. We present a video narrated by the senior author in which a description of the technique is offered. Thirty-eight patients underwent this approach to manage 40 lesions, including 15 dural arteriovenous fistulae, 9 arteriovenous malformations, 10 cavernous malformations, and 6 aneurysms. The extended retrosigmoid approach differs from the traditional approach with its C-shaped skin incision, posterior mastoidectomy, and extensive dissection of the sigmoid sinus, craniotomy rather than craniectomy, and anterior mobilization of the sinus with the dural flap. The application of the extended retrosigmoid approach to a series of complex lesions in the posterior fossa demonstrates its applicability as an alternative to radical cranial base approaches. The extended retrosigmoid approach requires a fundamental change in the management of the sigmoid sinus. The neurosurgeon must be familiar with petrous bone anatomy, experienced dissecting through bone using a high-speed drill, and comfortable working directly over a major venous sinus. The technical modifications of the extended retrosigmoid approach can be incorporated into the neurosurgical repertoire and will enhance exposure of the cerebellopontine angle and deep vascular structures, thereby minimizing the need for brain retraction and other transpetrous approaches.


Histological changes in the pituitary gland and adenomas following radiotherapy.
To elucidate histological changes in the pituitary gland and adenomas following radiotherapy, two irradiated pituitary glands and seven irradiated non-functioning adenomas were studied. The latter included four cases with conventional radiation (CR) and three cases with radiosurgery: two with gamma knife radiosurgery (GKR) and one with stereotactic fractionated radiotherapy (SRT). The specimens were obtained 10 months to 10 years (mean 58 months) after the radiotherapy. Irradiated pituitary glands showed diffuse fibrosis in the adenohypophysis, whereas irradiated adenomas showed either mild or no fibrosis in five CR/SRT cases and diffuse thick hyaline deposits in two GKR cases. No necrosis was observed. Stellate-shaped S-100 protein-positive cells were greater in number in the irradiated pituitary glands than in the normal glands. Pituitary cells with dense granular reactivity for mitochondrial protein, cytochrome oxidase, and Mn-SOD, mimicking oncocytes, were greater in number in the irradiated adenohypophysis but did not show any change in cell size. Many irradiated pituitary cells and some irradiated adenoma cells were densely positive with anticytokeratin 1,5,10,14 antibody whereas non-irradiated counterparts were negative. In adenomas, MIB-1 labeling index remained unchanged after the radiation. The results may indicate that radiation-induced fibrosis was associated with an increased number of folliculo-stellate cells, mitochondrial dysfunction, and squamous metaplasia. These findings were prominent in irradiated pituitary cells and may participate in delayed pituitary hypofunction following radiotherapy. In irradiated adenoma cells, similar findings were observed but diffuse fibrosis was absent. The histological changes were more intensive in adenomas following GKR than those following CR.


Hepatitis C virus infects rhesus macaque hepatocytes and simianized mice.
At least 170 million people are chronically infected with hepatitis C virus (HCV). Owing to the narrow host range of HCV and restricted use of chimpanzees, there is currently no suitable animal model for HCV pathogenesis studies or the development of a HCV vaccine. To identify cellular determinants of interspecies transmission and establish a novel immunocompetent model system, we examined the ability of HCV to infect hepatocytes from a small nonhuman primate, the rhesus macaque (Macaca mulatta). We show that the rhesus orthologs of critical HCV entry factors support viral glycoprotein-dependent virion uptake. Primary hepatocytes from rhesus macaques are also permissive for HCV-RNA replication and particle production, which is enhanced when antiviral signaling is suppressed. We demonstrate that this may be owing to the diminished capacity of HCV to antagonize mitochondrial antiviral-signaling protein-dependent innate cellular defenses. To test the ability of HCV to establish persistent replication in vivo, we engrafted primary rhesus macaque hepatocytes into immunocompromised xenorecipients. Inoculation of resulting simian liver chimeric mice with either HCV genotype 1a or 2a resulted in HCV serum viremia for up to 10 weeks. Together, these data indicate that rhesus macaques may be a viable model for HCV and implicate host immunity as a potential species-specific barrier to HCV infection. We conclude that suppression of host immunity or further viral adaptation may allow robust HCV infection in rhesus macaques and creation of a new animal model for studies of HCV pathogenesis, lentivirus coinfection, and vaccine development.


Measurement of Subfoveal Choroidal Thickness in Pseudoexfoliation Syndrome Using Enhanced Depth Imaging Optical Coherence Tomography.
Pseudoexfoliation (PSX) syndrome is associated with blood flow disturbances; however, its exact effect on choroidal blood flow and thickness remains to be elucidated. This study compared subfoveal choroidal thickness in normal eyes and in eyes with PSX using enhanced depth imaging optical coherence tomography (EDI-OCT). This prospective, cross-sectional study included 35 eyes of 35 patients (20 males, 15 females) with unilateral or bilateral PSX and 26 eyes of 26 healthy volunteers (13 males, 13 females). Besides a comprehensive ocular and physical examination, all subjects underwent EDI-OCT examination using an Optovue RTVue OCT device (Optovue Inc., Fremont, Calif., USA). The mean choroidal thickness (CT) and ocular perfusion pressure (OPP) were lower in the PSX group than in the healthy controls (249.4 ± 46.3 vs. 282.5 ± 55.8 µm, p = 0.014 and 40.7 ± 5.8 vs. 44.3 ± 4.3 mm Hg, p = 0.007, respectively). In addition, both systolic blood pressure and diastolic blood pressure measurements were lower among the PSX patients. However, no correlation was found between CT and OPP. The findings of this study suggest that PSX is associated with an overall thinning of the subfoveal choroid and a significant decrease in OPP. Future studies are warranted to further examine these relations.


Mapping the electrophysiological marker of sleep depth reveals skill maturation in children and adolescents.
Electroencephalographically (EEG) recorded slow wave activity (SWA, 1-4.5Hz), reflecting the depth of sleep, is suggested to play a crucial role in synaptic plasticity. Mapping of SWA by means of high-density EEG reveals that cortical regions showing signs of maturational changes (structural and behavioral) during childhood and adolescence exhibit more SWA. Moreover, the maturation of specific skills is predicted by the topographical distribution of SWA. Thus, SWA topography may serve as a promising neuroimaging tool with prognostic potential. Finally, our data suggest that deep sleep SWA in humans is involved in cortical development that optimizes performance.


Efficacy of Astaxanthin for the Treatment of Atopic Dermatitis in a Murine Model.
Atopic dermatitis (AD) is a common chronic inflammatory skin disease associated with various factors, including immunological abnormalities and exposure to allergens. Astaxanthin (AST) is a xanthophyll carotenoid that has recently been demonstrated to have anti-inflammatory effects and to regulate the expression of inflammatory cytokines. Thus, we investigated whether AST could improve the dermatitis and pruritus in a murine model of AD using NC/Nga mice. In addition to a behavioral evaluation, the effects of AST on the AD were determined by the clinical skin severity score, serum IgE level, histological analyses of skin, and by reverse transcription-PCR and Western blotting analyses for the expression of inflammation-related factors. AST (100 mg/kg) or vehicle (olive oil) was orally administered once day and three times a week for 26 days. When compared with vehicle-treated group, the administration of AST significantly reduced the clinical skin severity score. In addition, the spontaneous scratching in AD model mice was reduced by AST administration. Moreover, the serum IgE level was markedly decreased by the oral administration of AST compared to that in vehicle-treated mice. The number of eosinophils, total and degranulated mast cells all significantly decreased in the skin of AST-treated mice compared with vehicle-treated mice. The mRNA and protein levels of eotaxin, MIF, IL-4, IL-5 and L-histidine decarboxylase were significantly decreased in the skin of AST-treated mice compared with vehicle-treated mice. These results suggest that AST improves the dermatitis and pruritus in AD via the regulation of the inflammatory effects and the expression of inflammatory cytokines.


Odor-Specific Daily Rhythms in the Olfactory Sensitivity and Behavior of Aedes aegypti Mosquitoes.
Many biological processes and behaviors in mosquitoes display rhythmic patterns, allowing for fine tuning to cyclic environmental conditions. In mosquitoes, vector-host interactions are primarily mediated by olfactory signals. Previous studies have established that, in the malaria vector Anopheles gambiae, rhythmic expression of odorant binding proteins and takeout proteins in the antenna resulted in a corresponding rhythm in olfactory sensitivity to relevant host odors. However, it remained unclear how rhythms observed in olfactory sensitivity affect or explain rhythms in behavioral output, which ultimately impacts disease transmission. In order to address this knowledge gap, we quantified and compared patterns in locomotor activity, olfactory sensitivity, and olfactory behaviors in adult female Aedes aegypti mosquitoes. Here, we demonstrate an odorant-specific modulation of olfactory sensitivity in Ae. aegypti, decoupled from rhythms in olfactory behavior. Additionally, behavioral assays performed herein represent the first evidence of a time-dependence of the olfactory activation of behavior in Ae. aegypti mosquitoes. Results suggest that olfactory behavior of Aedes mosquitoes is modulated at both the peripheral (antenna) and central levels. As such, this work serves as a foundation for future studies aimed at further understanding the neural and molecular mechanisms underlying behavioral plasticity.


Fluorinated carbohydrates as chemical probes for molecular recognition studies. Current status and perspectives.
This review provides an extensive summary of the effects of carbohydrate fluorination with regard to changes in physical, chemical and biological properties with respect to regular saccharides. The specific structural, conformational, stability, reactivity and interaction features of fluorinated sugars are described, as well as their applications as probes and in chemical biology.


A new side effect of synthetic cannabinoid use by the bucket (waterpipe) method: Acute respiratory distress syndrome (ARDS).
The use and content of synthetic canibinden (SCs) has been rapidly increased in the last decades. The complex content of these substances bring along a wide spectrum of side effects. In addition to the expected neuropsychological side effects of pleasure-inducing substances such as agitation, anxiety, panic attack and hallucinations, rare cases of cerebrovascular diseases, seizures, acute renal injury, myocardial infarction and chronic lung injury have also been previously reported. Here we report a 19-year -old male who was presented with acute respiratory distress syndrome (ARDS) within hours of inhaled SC use with the rarely preferred bucket method. There is limited information in the literature about pulmonary effects of SCs and we could not detect any other ARDS case that develoed within hours after consumption of SC with the bucket method.


Occupational Exposure to Polycyclic Aromatic Hydrocarbon of Wildland Firefighters at Prescribed and Wildland Fires.
Wildland firefighters suppressing wildland fires or conducting prescribed fires work long shifts during which they are exposed to high levels of wood smoke with no respiratory protection. Polycyclic aromatic hydrocarbons (PAHs) are hazardous air pollutants formed during incomplete combustion. Exposure to PAHs was measured for 21 wildland firefighters suppressing two wildland fires and 4 wildland firefighters conducting prescribed burns in California. Personal air samples were actively collected using XAD4-coated quartz fiber filters and XAD2 sorbent tubes. Samples were analyzed for 17 individual PAHs through extraction with dichloromethane and gas chromatograph-mass spectrometer analysis. Naphthalene, retene, and phenanthrene were consistently the highest measured PAHs. PAH concentrations were higher at wildland fires compared to prescribed fires and were highest for firefighters during job tasks that involve the most direct contact with smoke near an actively burning wildland fire. Although concentrations did not exceed current occupational exposure limits, wildland firefighters are exposed to PAHs not only on the fire line at wildland fires, but also while working prescribed burns and while off-duty. Characterization of occupational exposures from wildland firefighting is important to understand better any potential long-term health effects.


Magnetic resonance imaging findings in 25 dogs with inflammatory cerebrospinal fluid.
To describe the signs that may be associated with intracranial inflammatory conditions, magnetic resonance (MR) images of 25 dogs that had inflammatory cerebrospinal fluid (CSF) were mixed with those of a control group of 40 dogs that had CSF negative for inflammatory disease and reviewed without knowledge of the clinical signs or diagnosis. CSF was considered inflammatory if the protein level was > 0.25 g/l and the white cell count was > 5 mm(-3). Abnormalities were found by MR imaging in 19 (76%) dogs with inflammatory CSF. Two dogs had focal lesions, 10 had multifocal lesions, and seven had diffuse lesions. Lesions affected all divisions of the brain. Mass effect was identified in seven (28%) dogs, including one that had a choroid plexus carcinoma. Lesions were hyperintense in T2-weighted images in 18 dogs and hypointense in T1-weighted images in six dogs. Multifocal or diffuse intraaxial lesions that were hyperintense in T2-weighted images were observed in 17 (68%) dogs with inflammatory CSF. Administration of gadolinium resulted in enhancement of intraaxial lesions in nine (36%) dogs and enhancement of meninges in seven (28%) dogs. Six (24%) dogs with inflammatory CSF had images interpreted as normal.


Protein Misfolding Diseases and Therapeutic Approaches.
Protein folding is the process by which a polypeptide chain acquires its functional, native 3D structure. Protein misfolding, on the other hand, is a process in which protein fails to fold into its native functional conformation. This misfolding of proteins may lead to precipitation of a number of serious diseases such as Cystic Fibrosis (CF), Alzheimer's Disease (AD), Parkinson's Disease (PD), and Amyotrophic Lateral Sclerosis (ALS) etc. Protein Quality-control (PQC) systems, consisting of molecular chaperones, proteases and regulatory factors, help in protein folding and prevent its aggregation. At the same time, PQC systems also do sorting and removal of improperly folded polypeptides. Among the major types of PQC systems involved in protein homeostasis are cytosolic, Endoplasmic Reticulum (ER) and mitochondrial ones. The cytosol PQC system includes a large number of component chaperones, such as Nascent-polypeptide-associated Complex (NAC), Hsp40, Hsp70, prefoldin and T Complex Protein-1 (TCP-1) Ring Complex (TRiC). Protein misfolding diseases caused due to defective cytosolic PQC system include diseases involving keratin/collagen proteins, cardiomyopathies, phenylketonuria, PD and ALS. The components of PQC system of Endoplasmic Reticulum (ER) include Binding immunoglobulin Protein (BiP), Calnexin (CNX), Calreticulin (CRT), Glucose-regulated Protein GRP94, the thiol-disulphide oxidoreductases, Protein Disulphide Isomerase (PDI) and ERp57. ER-linked misfolding diseases include CF and Familial Neurohypophyseal Diabetes Insipidus (FNDI). The components of mitochondrial PQC system include mitochondrial chaperones such as the Hsp70, the Hsp60/Hsp10 and a set of proteases having AAA+ domains similar to the proteasome that are situated in the matrix or the inner membrane. Protein misfolding diseases caused due to defective mitochondrial PQC system include medium-chain acyl-CoA dehydrogenase (MCAD)/Short-chain Acyl-CoA Dehydrogenase (SCAD) deficiency diseases, hereditary spastic paraplegia. Among therapeutic approaches towards the treatment of various protein misfolding diseases, chaperones have been suggested as potential therapeutic molecules for target based treatment. Chaperones have been advantageous because of their efficient entry and distribution inside the cells, including specific cellular compartments, in therapeutic concentrations. Based on the chemical nature of the chaperones used for therapeutic purposes, molecular, chemical and pharmacological classes of chaperones have been discussed.


Use of basal insulin and the associated clinical outcomes among elderly nursing home residents with type 2 diabetes mellitus: a retrospective chart review study.
The management of type 2 diabetes mellitus in long-term care (LTC) settings can be complex as a result of age-related complications. Despite guideline recommendations, sliding scale insulin remains commonplace in the LTC setting and data on basal insulin use are lacking. This retrospective study used medical chart data and the Minimum Data Set from elderly LTC facility patients who received basal insulin (insulin glargine, insulin detemir, or neutral protamine Hagedorn insulin) for the treatment of diabetes, to investigate the practice patterns and associated clinical outcomes. A total of 2,096 elderly, insulin-treated patients in LTC were identified, with 59.5% of them (N=1,247) receiving basal insulin. Of these, more than 50% of patients received sliding scale insulin in co-administration with basal insulin. Despite its ease of use, insulin pen use was very low, at 14.6%. Significant differences were observed between the basal insulin groups for glycated hemoglobin level and dosing frequency. Hypoglycemia was uncommon -17.2% of patients experienced at least one event, and there was no significant difference in the prevalence of hypoglycemia between the groups. These data suggest the underutilization of basal insulin in the LTC setting and worryingly high combinational use with sliding scale insulin. Differences in glycated hemoglobin and dosing frequencies between types of basal insulin warrant further comparative effectiveness studies.


Comparison of fluoroscopy time during coronary angiography and interventions by radial and femoral routes- can we decrease the fluoroscopy time with increased experience? An observational study.
Radial route of access is increasingly being used for coronary angiograms and intervention. However, radiation exposure of operators was not known in our set up with either transfemoral or transradial procedures. The objective of the study was to compare related peripheral arterial route radiation exposure of operators by assessing fluoroscopy time. The secondary objective was to determine the relationship of operator experience with fluoroscopy time. This observational study was conducted in a tertiary care center - Cardiovascular Institute of Karachi (Pakistan) during the period of July 1(st) 2009 to September 30(th) 2009. We studied 1016 consecutive adult patients referred for coronary angiography (CA) or percutaneous coronary intervention (PCI). Patients who underwent right heart catheterization or for valvuloplasty were excluded from the study. Out of these 1016 patients, 928 were diagnostic CAs (734 via femoral route [f-CA] and 194 via radial route [r-CA]) and 88 were PCI (64 via femoral route [f-PCI] and 24 via radial route [r-PCI]). Fluoroscopy time was recorded as a surrogate of radiation exposure. Statistical analysis was performed using unpaired t, Mann-Whitney U, Chi-square and ANOVA tests. Mean fluoroscopy time was found to be significantly higher in patients who underwent r-CA (6.3±3.8 vs 4.0±2.9 min; p<0.001) and r-PCI (15.1±11.8 vs 10.3±7.4 min; p=0.02) as compared with those underwent f-CA and f-PCI. Mean fluoroscopy time of well experienced operators was also high in r-CAs (5.4±2.9 vs 4.2±3.5 min; p=0.004). Radial procedures are associated with longer fluoroscopy time that may result in high radiation exposure to radial operators. Even well experienced radial operators cannot minimize their fluoroscopy time to the level of well experienced femoral operators.


Maternal antibiotic-induced early changes in microbial colonization selectively modulate colonic permeability and inducible heat shock proteins, and digesta concentrations of alkaline phosphatase and TLR-stimulants in swine offspring.
Elevated intake of high energy diets is a risk factor for the development of metabolic diseases and obesity. High fat diets cause alterations in colonic microbiota composition and increase gut permeability to bacterial lipopolysaccharide, and subsequent low-grade chronic inflammation in mice. Chronic inflammatory bowel diseases are increasing worldwide and may involve alterations in microbiota-host dialog. Metabolic disorders appearing in later life are also suspected to reflect changes in early programming. However, how the latter affects the colon remains poorly studied. Here, we hypothesized that various components of colonic physiology, including permeability, ion exchange and protective inducible heat shock proteins (HSP) are influenced in the short- and long-terms by early disturbances in microbial colonization. The hypothesis was tested in a swine model. Offspring were born to control mothers (n = 12) or mothers treated with the antibiotic (ATB) amoxicillin around parturition (n = 11). Offspring were slaughtered between 14 and 42 days of age to study short-term effects. For long-term effects, young adult offspring from the same litters consumed a normal or a palm oil-enriched diet for 4 weeks between 140 and 169 days of age. ATB treatment transiently modified maternal fecal microbiota although the minor differences observed for offspring colonic microbiota were nonsignificant. In the short-term, consistently higher HSP27 and HSP70 levels and transiently increased horseradish peroxidase permeability in ATB offspring colon were observed. Importantly, long-term consequences included reduced colonic horseradish peroxidase permeability, and increased colonic digesta alkaline phosphatase (AP) and TLR2- and TLR4-stimulant concentrations in rectal digesta in adult ATB offspring. Inducible HSP27 and HSP70 did not change. Interactions between early ATB treatment and later diet were noted for paracellular permeability and concentrations of colonic digesta AP. In conclusion, our data suggest that early ATB-induced changes in bacterial colonization modulate important aspects of colonic physiology in the short- and long-terms.


Rapid localized crystallization of lysozyme by laser trapping.
Confining protein crystallization to a millimetre size was achieved within 0.5 h after stopping 1 h intense trapping laser irradiation, which shows excellent performance in spatial and temporal controllability compared to spontaneous nucleation. A continuous-wave near-infrared laser beam is tightly focused into a glass/solution interfacial layer of a supersaturated buffer solution of hen egg-white lysozyme (HEWL). The crystallization is not observed during laser trapping, but initiated by stopping the laser irradiation. The generated crystals are localized densely in a circular area with a diameter of a few millimetres around the focal spot and show specific directions of the optical axes of the HEWL crystals. To interpret this unique crystallization, we propose a mechanism that nucleation and the subsequent growth take place in a highly concentrated domain consisting of HEWL liquid-like clusters after turning off laser trapping.


Genetic analysis and identification of SSR markers associated with rice blast disease in a BC2F1 backcross population.
Rice (Oryza sativa L.) blast disease is one of the most destructive rice diseases in the world. The fungal pathogen, Magnaporthe oryzae, is the causal agent of rice blast disease. Development of resistant cultivars is the most preferred method to achieve sustainable rice production. However, the effectiveness of resistant cultivars is hindered by the genetic plasticity of the pathogen genome. Therefore, information on genetic resistance and virulence stability are vital to increase our understanding of the molecular basis of blast disease resistance. The present study set out to elucidate the resistance pattern and identify potential simple sequence repeat markers linked with rice blast disease. A backcross population (BC2F1), derived from crossing MR264 and Pongsu Seribu 2 (PS2), was developed using marker-assisted backcross breeding. Twelve microsatellite markers carrying the blast resistance gene clearly demonstrated a polymorphic pattern between both parental lines. Among these, two markers, RM206 and RM5961, located on chromosome 11 exhibited the expected 1:1 testcross ratio in the BC2F1 population. The 195 BC2F1 plants inoculated against M. oryzae pathotype P7.2 showed a significantly different distribution in the backcrossed generation and followed Mendelian segregation based on a single-gene model. This indicates that blast resistance in PS2 is governed by a single dominant gene, which is linked to RM206 and RM5961 on chromosome 11. The findings presented in this study could be useful for future blast resistance studies in rice breeding programs.


Structural studies of bacterioferritin B from Pseudomonas aeruginosa suggest a gating mechanism for iron uptake via the ferroxidase center .
The structure of recombinant Pseudomonas aeruginosa bacterioferritin B (Pa BfrB) has been determined from crystals grown from protein devoid of core mineral iron (as-isolated) and from protein mineralized with approximately 600 iron atoms (mineralized). Structures were also obtained from crystals grown from mineralized BfrB after they had been soaked in an FeSO(4) solution (Fe soak) and in separate experiments after they had been soaked in an FeSO(4) solution followed by a soak in a crystallization solution (double soak). Although the structures consist of a typical bacterioferritin fold comprised of a nearly spherical 24-mer assembly that binds 12 heme molecules, comparison of microenvironments observed in the distinct structures provided interesting insights. The ferroxidase center in the as-isolated, mineralized, and double-soak structures is empty. The ferroxidase ligands (except His130) are poised to bind iron with minimal conformational changes. The His130 side chain, on the other hand, must rotate toward the ferroxidase center to coordinate iron. In comparison, the structure obtained from crystals soaked in an FeSO(4) solution displays a fully occupied ferroxidase center and iron bound to the internal, Fe((in)), and external, Fe((out)), surfaces of Pa BfrB. The conformation of His130 in this structure is rotated toward the ferroxidase center and coordinates an iron ion. The structures also revealed a pore on the surface of Pa BfrB that likely serves as a port of entry for Fe(2+) to the ferroxidase center. On its opposite end, the pore is capped by the side chain of His130 when it adopts its "gate-closed" conformation that enables coordination to a ferroxidase iron. A change to its "gate-open", noncoordinative conformation creates a path for the translocation of iron from the ferroxidase center to the interior cavity. These structural observations, together with findings obtained from iron incorporation measurements in solution, suggest that the ferroxidase pore is the dominant entry route for the uptake of iron by Pa BfrB. These findings, which are clearly distinct from those made with Escherichia coli Bfr [Crow, A. C., Lawson, T. L., Lewin, A., Moore, G. R., and Le Brun, N. E. (2009) J. Am. Chem. Soc. 131, 6808-6813], indicate that not all bacterioferritins operate in the same manner.


Mechanically induced silyl ester cleavage under acidic conditions investigated by AFM-based single-molecule force spectroscopy in the force-ramp mode.
AFM-based dynamic single-molecule force spectroscopy was used to stretch carboxymethylated amylose (CMA) polymers, which have been covalently tethered between a silanized glass substrate and a silanized AFM tip via acid-catalyzed ester condensation at pH 2.0. Rupture forces were measured as a function of temperature and force loading rate in the force-ramp mode. The data exhibit significant statistical scattering, which is fitted with a maximum likelihood estimation (MLE) algorithm. Bond rupture is described with a Morse potential based Arrhenius kinetics model. The fit yields a bond dissociation energy De = 35 kJ mol(-1) and an Arrhenius pre-factor A = 6.6 × 10(4) s(-1). The bond dissociation energy is consistent with previous experiments under identical conditions, where the force-clamp mode was employed. However, the bi-exponential decay kinetics, which the force-clamp results unambiguously revealed, are not evident in the force-ramp data. While it is possible to fit the force-ramp data with a bi-exponential model, the fit parameters differ from the force-clamp experiments. Overall, single-molecule force spectroscopy in the force-ramp mode yields data whose information content is more limited than force-clamp data. It may, however, still be necessary and advantageous to perform force-ramp experiments. The number of successful events is often higher in the force-ramp mode, and competing reaction pathways may make force-clamp experiments impossible.


Evasive strategies in rabies virus infection.
Rabies virus (RABV) is a strictly neurotropic virus that slowly propagates in the nervous system (NS) of the infected host from the site of entry (usually due to a bite) up to the site of exit (salivary glands). Successful achievement of the virus cycle relies on the preservation of the neuronal network. Once RABV has entered the NS, its progression is not interrupted either by destruction of the infected neurons or by the immune response, which are major host mechanisms for combating viral infection. RABV has developed two main mechanisms to escape the host defenses: (1) its ability to kill protective migrating T cells and (2) its ability to sneak into the NS without triggering apoptosis of the infected neurons and preserving the integrity of neurites.


Evaluation of three commercially available real-time PCR based systems for detection of Cronobacter species.
In the last few years, various PCR based methods have been developed that enable detection of Cronobacter spp. to the genus and species level. Moreover, several real-time PCR based systems for detection of Cronobacter spp. are available, however, comparative evaluation studies are not available. The current study represents a comparative evaluation of three commercial diagnostic systems, namely the BAX® System PCR Assay Enterobacter sakazakii (DuPont, Qualicon, Wilmington, USA), the Assurance GDS™ Enterobacter sakazakii (BioControl, Bellvue, USA) and the foodproof® Enterobacter sakazakii Detection Kit (Biotecon Diagnostics, Potsdam, Germany) for the rapid identification of Cronobacter spp. Twenty-one target and non-target strains were included in the study and results were compared for specificity and convenience in performance. A specificity of 100% was observed for two of the three real time PCR systems tested, namely the Assurance GDS™ Enterobacter sakazakii and the foodproof® Enterobacter sakazakii Detection Kit for pure cultures as well as artificially contaminated powdered infant formula (PIF) samples.


A conformational switch in Escherichia coli 16S ribosomal RNA during decoding of messenger RNA.
Direct evidence is presented for a conformational switch in 16S ribosomal RNA (rRNA) that affects tRNA binding to the ribosome and decoding of messenger RNA (mRNA). These data support the hypothesis that dynamic changes in rRNA structure occur during translation. The switch involves two alternating base-paired arrangements apparently facilitated by ribosomal proteins S5 and S12, and produces significant changes in the rRNA structure. Chemical probing shows reciprocal enhancements or protections at sites in 16S rRNA that are at or very near sites that were previously crosslinked to mRNA. These data indicate that the switch affects codon-anticodon arrangement and proper selection of tRNA at the ribosomal A site, and that the switch is a fundamental mechanism in all ribosomes.


Temporal profiles of viral load in posterior oropharyngeal saliva samples and serum antibody responses during infection by SARS-CoV-2: an observational cohort study.
Coronavirus disease 2019 (COVID-19) causes severe community and nosocomial outbreaks. Comprehensive data for serial respiratory viral load and serum antibody responses from patients infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are not yet available. Nasopharyngeal and throat swabs are usually obtained for serial viral load monitoring of respiratory infections but gathering these specimens can cause discomfort for patients and put health-care workers at risk. We aimed to ascertain the serial respiratory viral load of SARS-CoV-2 in posterior oropharyngeal (deep throat) saliva samples from patients with COVID-19, and serum antibody responses. We did a cohort study at two hospitals in Hong Kong. We included patients with laboratory-confirmed COVID-19. We obtained samples of blood, urine, posterior oropharyngeal saliva, and rectal swabs. Serial viral load was ascertained by reverse transcriptase quantitative PCR (RT-qPCR). Antibody levels against the SARS-CoV-2 internal nucleoprotein (NP) and surface spike protein receptor binding domain (RBD) were measured using EIA. Whole-genome sequencing was done to identify possible mutations arising during infection. Between Jan 22, 2020, and Feb 12, 2020, 30 patients were screened for inclusion, of whom 23 were included (median age 62 years [range 37-75]). The median viral load in posterior oropharyngeal saliva or other respiratory specimens at presentation was 5·2 log10 copies per mL (IQR 4·1-7·0). Salivary viral load was highest during the first week after symptom onset and subsequently declined with time (slope -0·15, 95% CI -0·19 to -0·11; R2=0·71). In one patient, viral RNA was detected 25 days after symptom onset. Older age was correlated with higher viral load (Spearman's ρ=0·48, 95% CI 0·074-0·75; p=0·020). For 16 patients with serum samples available 14 days or longer after symptom onset, rates of seropositivity were 94% for anti-NP IgG (n=15), 88% for anti-NP IgM (n=14), 100% for anti-RBD IgG (n=16), and 94% for anti-RBD IgM (n=15). Anti-SARS-CoV-2-NP or anti-SARS-CoV-2-RBD IgG levels correlated with virus neutralisation titre (R2>0·9). No genome mutations were detected on serial samples. Posterior oropharyngeal saliva samples are a non-invasive specimen more acceptable to patients and health-care workers. Unlike severe acute respiratory syndrome, patients with COVID-19 had the highest viral load near presentation, which could account for the fast-spreading nature of this epidemic. This finding emphasises the importance of stringent infection control and early use of potent antiviral agents, alone or in combination, for high-risk individuals. Serological assay can complement RT-qPCR for diagnosis. Richard and Carol Yu, May Tam Mak Mei Yin, The Shaw Foundation Hong Kong, Michael Tong, Marina Lee, Government Consultancy Service, and Sanming Project of Medicine.


Acid-base adjustments and first evidence of denticle corrosion caused by ocean acidification conditions in a demersal shark species.
Global ocean acidification is expected to chronically lower the pH to 7.3 (>2200 µatm seawater pCO2) by the year 2300. Acute hypercapnia already occurs along the South African west and south coasts due to upwelling- and low-oxygen events, with increasing frequency. In the present project we investigated the impact of hypercapnia on the endemic demersal shark species Haploblepharus edwardsii. Specifically, we experimentally analysed acid-base regulation during acute and chronic hypercapnia, the effects of chronic hypercapnia on growth rates and on denticle structure- and composition. While H. edwardsii are physiologically well adapted to acute and chronic hypercapnia, we observed, for the first time, denticle corrosion as a result of chronic exposure. We conclude that denticle corrosion could increase denticle turnover and compromise hydrodynamics and skin protection.


Detecting and measuring small numbers of viable Coxiella burnetii.
Coxiella burnetii is an acidophilic, intracellular bacterium that causes the human disease Q fever. In some studies, it is important to distinguish between viable and nonviable C. burnetii. We compared four methods for detecting and measuring viable C. burnetii in biological samples as follows: growth in two different cell culture lines, infection of severe combined immunodeficient (SCID) mice (leading to death) and infection of SCID mice with detection of C. burnetii in their spleen (after euthanasia at day 50 postinfection). Two isolates of C. burnetii were used ('Henzerling' and 'Arandale'). Our in-house qPCR assay for C. burnetii DNA was used as a control. SCID mouse inoculation was more sensitive than cell culture. The assay that detected C. burnetii in SCID mouse spleens was slightly more sensitive than SCID mice deaths alone. Approximately one viable C. burnetii cell could be detected by this method, making it suitable for determining the viability of C. burnetii in a sample.


Lung donor selection criteria.
The criteria that define acceptable physiologic and social parameters for lung donation have remained constant since their empiric determination in the 1980s. These criteria include a donor age between 25-40, a arterial partial pressure of oxygen (PaO2)/FiO2 ratio greater than 350, no smoking history, a clear chest X-ray, clean bronchoscopy, and a minimal ischemic time. Due to the paucity of organ donors, and the increasing number of patients requiring lung transplant, finding a donor that meets all of these criteria is quite rare. As such, many transplants have been performed where the donor does not meet these stringent criteria. Over the last decade, numerous reports have been published examining the effects of individual acceptance criteria on lung transplant survival and graft function. These studies suggest that there is little impact of the historical criteria on either short or long term outcomes. For age, donors should be within 18 to 64 years old. Gender may relay benefit to all female recipients especially in male to female transplants, although results are mixed in these studies. Race matched donor/recipients have improved outcomes and African American donors convey worse prognosis. Smoking donors may decrease recipient survival post transplant, but provide a life saving opportunity for recipients that may otherwise remain on the transplant waiting list. No specific gram stain or bronchoscopic findings are reflected in recipient outcomes. Chest radiographs are a poor indicator of lung donor function and should not adversely affect organ usage aside for concerns over malignancy. Ischemic time greater than six hours has no documented adverse effects on recipient mortality and should not limit donor retrieval distances. Brain dead donors and deceased donors have equivalent prognosis. Initial PaO2/FiO2 ratios less than 300 should not dissuade donor organ usage, although recruitment techniques should be implemented with intent to transplant.


An Outbreak of Onion Center Rot Caused by Pantoea ananatis in Korea.
Symptoms typical of center rot of onion (Allium cepa L.) were observed in farmers' fields in spring of 2009 and 2010 in Hamyang, South Korea, at an incidence of 30 to 50% (five affected fields representing approximately 6 ha). The symptoms were identical to those reported from infected onions in Georgia in 1997 (1). Harvested bulbs of symptomatic plants had reddish, collapsed scales near the neck. Symptomatic bulb tissues were surface-sterilized by immersing sections of the tissue for 30 s in 1% NaOCl, then rinsing the sections with sterilized, distilled water. Tissues were then macerated in 1 ml of sterilized, distilled water in a 1.5 ml Eppendorf tube using a sterile scalpel. The macerated tissue was left to soak for 10 min, after which a 5 μl suspension from each section was streaked onto plates of nutrient agar (NA). Gram-negative, rod-shaped, yellow bacteria were consistently recovered on NA. Three bacterial isolates recovered were each facultative anaerobes and induced a hypersensitive reaction on tobacco leaves. The biochemical test, API 20E (Biomérieux, Marcy l'Etoile, France), was also used for identification. All three strains tested positive for β-D-galactosidase, utilization of citrate, and production of acetoin, catalase, and indole. All three strains tested negative for ornithine decarboxylase, lysine decarboxylase, urease, and oxidase. All produced acid from arabinose, glucose, mannitol, and sorbitol; while none produced acid from melibiose, inositol, and rhamnose. These characteristics are consistent with those of P. ananatis (1,2). Five bulbs were each surface-disinfested with 70% ethanol, dried, and injected with 50 μl of the appropriate bacterial suspension containing ~108 CFU/ml, using a syringe. The bulbs were placed in plastic boxes with four sheets of wet paper towel to maintain the relative humidity at 100%, and incubated at 25°C for 2 weeks. Three onion bulbs treated similarly but injected with sterilized, distilled water served as replicates of the control treatment. After 1 week of incubation, inoculated onion bulbs developed a brown discoloration and decay of the internal, fleshy scales matching those observed in the original farmers' fields. The control onion bulbs remained asymptomatic. Bacteria reisolated from lesions in the fleshy bulb scales of the inoculated bulbs had the same characteristics as the original isolates inoculated, proving Koch's postulates. Bacteria were not reisolated from any of the control bulbs. To confirm identity of the isolated bacteria, 16S rRNA and recA genes were amplified with primers 27mF: 5'-AGAGTTTGATCMTGGCTCAG-3' and 1492mR: 5'-GGYTACCTTGTTACGACTT-3', and PAGRECA21: 5'-GGTGAAGACCGCTCAATGGA-3' and PAGRECA621: 5'-CACCGATACGGCGGATATCA-3', respectively (3). Amplification of the 16S rRNA gene generated a 1,506-bp consensus sequence (GenBank Accession No. JQ762264), and amplification of the recA gene generated a consensus sequence of 601 bp (JQ762265). The 16S rRNA and recA gene sequences shared 99% nucleotide identity with those of a P. ananatis strain in GenBank (DQ195523 and AY219004, respectively). Based on symptoms, biochemical tests, and molecular analyses, the bacterium responsible for the onion symptoms in Korea was identified as P. ananatis. To our knowledge, this is the first report of center rot of onion caused by P. ananatis in Korea. References: (1) R. D. Gitaitis and J. D. Gay. Plant Dis. 81:1096, 1997. (2) H. G. Truper and L. de Clari. Int. J. Syst. Bacteriol. 47:908, 1997. (3) A. Wensing et al. Appl. Environ. Microbiol. 76:6248, 2010.


Associations between diet and disease activity in ulcerative colitis patients using a novel method of data analysis.
The relapsing nature and varying geographical prevalence of ulcerative colitis (UC) implicates environmental factors such as diet in its aetiology. In order to determine which foods might be related to disease activity in UC a new method of dietary analysis was developed and applied. Eighty-one UC patients were recruited at all stages of the disease process. Following completion of a 7 d diet diary, clinical assessment including a sigmoidoscopic examination (scale 0 (normal mucosa) to 6 (very active disease)) was conducted. Food weights for each person were adjusted (divided) by the person's calorific intake for the week. Each food consumed was given a food sigmoidoscopy score (FSS) calculated by summing the products of the (adjusted) weight of food consumed and sigmoidoscopy score for each patient and occurrence of food and dividing by the total (adjusted) weight of the food consumed by all 81 patients. Thus, foods eaten in large quantities by patients with very active disease have high FSSs and vice versa. Foods consumed by <10 people or weighing <1 kg for the whole group were excluded, leaving 75 foods. High FSS foods were characterized by high levels of the anti-thiamin additive sulfite (Mann-Whitney, p < 0.001), i.e. bitter, white wine, burgers, soft drinks from concentrates, sausages, lager and red wine. Caffeine also has anti-thiamin properties and decaffeinated coffee was associated with a better clinical state than the caffeine containing version. Beneficial foods (average intake per week) included pork (210 g), breakfast cereals (200 g), lettuce (110 g), apples and pears (390 g), milk (1250 ml), melon (350 g), bananas (350 g), bacon (120 g), beef and beef products (500 g), tomatoes (240 g), soup (700 g), citrus fruits (300 g), fish (290 g), yogurt (410 g), cheese (110 g), potatoes (710 g) and legumes (120 g). The dietary analysis method described provides a new tool for establishing relationships between diet and disease and indicates a potentially therapeutic diet for UC.


Solving Nonlinearly Separable Classifications in a Single-Layer Neural Network.
Since the work of Minsky and Papert ( 1969 ), it has been understood that single-layer neural networks cannot solve nonlinearly separable classifications (i.e., XOR). We describe and test a novel divergent autoassociative architecture capable of solving nonlinearly separable classifications with a single layer of weights. The proposed network consists of class-specific linear autoassociators. The power of the model comes from treating classification problems as within-class feature prediction rather than directly optimizing a discriminant function. We show unprecedented learning capabilities for a simple, single-layer network (i.e., solving XOR) and demonstrate that the famous limitation in acquiring nonlinearly separable problems is not just about the need for a hidden layer; it is about the choice between directly predicting classes or learning to classify indirectly by predicting features.


Incongruence Between Observers' and Observed Facial Muscle Activation Reduces Recognition of Emotional Facial Expressions From Video Stimuli.
According to embodied cognition accounts, viewing others' facial emotion can elicit the respective emotion representation in observers which entails simulations of sensory, motor, and contextual experiences. In line with that, published research found viewing others' facial emotion to elicit automatic matched facial muscle activation, which was further found to facilitate emotion recognition. Perhaps making congruent facial muscle activity explicit produces an even greater recognition advantage. If there is conflicting sensory information, i.e., incongruent facial muscle activity, this might impede recognition. The effects of actively manipulating facial muscle activity on facial emotion recognition from videos were investigated across three experimental conditions: (a) explicit imitation of viewed facial emotional expressions (stimulus-congruent condition), (b) pen-holding with the lips (stimulus-incongruent condition), and (c) passive viewing (control condition). It was hypothesised that (1) experimental condition (a) and (b) result in greater facial muscle activity than (c), (2) experimental condition (a) increases emotion recognition accuracy from others' faces compared to (c), (3) experimental condition (b) lowers recognition accuracy for expressions with a salient facial feature in the lower, but not the upper face area, compared to (c). Participants (42 males, 42 females) underwent a facial emotion recognition experiment (ADFES-BIV) while electromyography (EMG) was recorded from five facial muscle sites. The experimental conditions' order was counter-balanced. Pen-holding caused stimulus-incongruent facial muscle activity for expressions with facial feature saliency in the lower face region, which reduced recognition of lower face region emotions. Explicit imitation caused stimulus-congruent facial muscle activity without modulating recognition. Methodological implications are discussed.


Curcumin confers protection to irradiated THP-1 cells while its nanoformulation sensitizes these cells via apoptosis induction.
Protection against ionizing radiation (IR) and sensitization of cancer cells to IR are apparently contrasting phenomena. However, curcumin takes on these contrasting roles leading to either protection or enhanced apoptosis in different irradiated cells. Here we studied whether pretreatment with free curcumin or a novel dendrosomal nanoformulation of curcumin (DNC) could exert protective/sensitizing effects on irradiated THP-1 leukemia cells. We employed assays including MTT viability, clonogenic survival, DNA fragmentation, PI/Annexin V flow cytometry, antioxidant system (ROS, TBARS for lipid peroxidation, 8-OHdG and γH2AX for DNA damage, glutathione, CAT and GPx activity, enzymes gene expression), ELISA (NF-κB and Nrf2 binding, TNF-α release), caspase assay, siRNA silencing of caspase-3, and western blotting to illustrate the observed protective role of curcumin in comparison with the opposite sensitizing role of its nanoformulation at a similar 10 μM concentration. The in vivo relevance of this concentration was determined via intraperitoneal administration in mice. Curcumin significantly enhanced the antioxidant defense, while DNC induced apoptosis and reduced viability as well as survival of irradiated THP-1 cells. Nrf2 binding showed an early rise and fall in DNC-treated cells, despite a gradual increase in curcumin-treated cells. We also demonstrated that DNC induced apoptosis in THP-1 cells via caspase-3 activation; whereas in combination with radiation, DNC alternatively employed a caspase-independent apoptosis pathway involving cytochrome c release from mitochondria.


The protective mechanisms underlying Ginsenoside Rg1 effects on rat sciatic nerve injury.
Ginsenoside Rg1 (GsRg1), derived from the herb Ginseng, was found to exert protective effects in nerve injury; however, the mechanisms underlying these effects remain to be determined. Oxidant stress and apoptosis are known to be involved in sciatic nerve injury. Thus, the aim of this study was to examine whether GsRg1 was able to modify sciatic nerve injury in a rat model. The following parameters were measured: (1) number of spinal cord motoneurons by Nissl staining, (2) oxidation parameters including spinal cord malondialdehyde (MDA) levels and activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) as well as (3) involvement of apoptosis by determining caspase-3 and X-linked inhibitor of apoptosis protein (XIAP) by immunohistochemistry and Western blot. The number of spinal cord motoneurons was significantly reduced after sciatic nerve injury, while treatment with GsRg1 markedly elevated cell number. Sciatic nerve injury markedly increased spinal cord MDA content concomitant with reduced activities of SOD and GSH-Px. GsRg1 significantly decreased MDA content accompanied by elevated activities of SOD and GSH-Px. Further nerve injury significantly diminished protein expression levels of XIAP accompanied by elevated protein expression levels of caspase-3 in the spinal cord. GsRg1 markedly increased protein expression levels of XIAP, but significantly reduced protein expression levels of caspase-3. Data suggest that the protective effects of GsRg1 in sciatic nerve injury may be associated with reduced oxidative stress involving anti-apoptotic pathways.


Does minimally invasive lumbar disc surgery result in less muscle injury than conventional surgery? A randomized controlled trial.
The concept of minimally invasive lumbar disc surgery comprises reduced muscle injury. The aim of this study was to evaluate creatine phosphokinase (CPK) in serum and the cross-sectional area (CSA) of the multifidus muscle on magnetic resonance imaging as indicators of muscle injury. We present the results of a double-blind randomized trial on patients with lumbar disc herniation, in which tubular discectomy and conventional microdiscectomy were compared. In 216 patients, CPK was measured before surgery and at day 1 after surgery. In 140 patients, the CSA of the multifidus muscle was measured at the affected disc level before surgery and at 1 year after surgery. The ratios (i.e. post surgery/pre surgery) of CPK and CSA were used as outcome measures. The multifidus atrophy was classified into three grades ranging from 0 (normal) to 3 (severe atrophy), and the difference between post and pre surgery was used as an outcome. Patients' low-back pain scores on the visual analogue scale (VAS) were documented before surgery and at various moments during follow-up. Tubular discectomy compared with conventional microdiscectomy resulted in a nonsignificant difference in CPK ratio, although the CSA ratio was significantly lower in tubular discectomy. At 1 year, there was no difference in atrophy grade between both groups nor in the percentage of patients showing an increased atrophy grade (14% tubular vs. 18% conventional). The postoperative low-back pain scores on the VAS improved in both groups, although the 1-year between-group mean difference of improvement was 3.5 mm (95% CI; 1.4-5.7 mm) in favour of conventional microdiscectomy. In conclusion, tubular discectomy compared with conventional microdiscectomy did not result in reduced muscle injury. Postoperative evaluation of CPK and the multifidus muscle showed similar results in both groups, although patients who underwent tubular discectomy reported more low-back pain during the first year after surgery.


Systematic Review and Meta-Analysis of Peer-Led Self-Management Programs for Increasing Physical Activity.
Approximately 85 % of Canadians are not physically active enough to achieve health benefits. Peer-led self-management programs are becoming an increasingly popular strategy for modifying health behaviors, including physical activity. The purpose of this study was to systematically review and meta-analyze the effect of peer-led self-management interventions on physical activity. PubMed, MEDLINE, PsycINFO, EMBASE, CINAHL, and Cochrane Database of Systematic Reviews databases were systematically searched to identify all relevant randomized controlled trials that evaluated the effect of peer-led self-management on physical activity. The studies were described and effect size data were included in meta-analyses. Subgroup analyses were performed according to type of physical activity outcome (i.e., duration, frequency, other). Twenty-one studies were included in the review and 14 reported statistically significant improvements in physical activity. A meta-analysis of 17 studies showed a statistically significant moderate pooled effect (standardized mean difference (SMD) = 0.4, p < 0.001) of peer-led self-management programs on physical activity immediately post-intervention. The intervention had a large statistically significant effect based on the four studies that included follow-up measures (SMD = 1.5, p = 0.03). Meta-analysis of nine studies that used similar outcomes (i.e., minutes of physical activity) revealed a statistically significant small effect (SMD = 0.2, p < 0.001). Peer-led self-management programs appear to be effective at increasing weekly duration of physical activity in various populations, but the effect size is small. Training peers to encourage increased physical activity may provide an effective method for reaching various clinical and non-clinical populations. More research is needed using validated and consistent physical activity outcomes.


A quantitative study of the division cycle of Caulobacter crescentus stalked cells.
Progression of a cell through the division cycle is tightly controlled at different steps to ensure the integrity of genome replication and partitioning to daughter cells. From published experimental evidence, we propose a molecular mechanism for control of the cell division cycle in Caulobacter crescentus. The mechanism, which is based on the synthesis and degradation of three "master regulator" proteins (CtrA, GcrA, and DnaA), is converted into a quantitative model, in order to study the temporal dynamics of these and other cell cycle proteins. The model accounts for important details of the physiology, biochemistry, and genetics of cell cycle control in stalked C. crescentus cell. It reproduces protein time courses in wild-type cells, mimics correctly the phenotypes of many mutant strains, and predicts the phenotypes of currently uncharacterized mutants. Since many of the proteins involved in regulating the cell cycle of C. crescentus are conserved among many genera of alpha-proteobacteria, the proposed mechanism may be applicable to other species of importance in agriculture and medicine.


Internal fixation of a femur fracture in an American bullfrog.
An adult male American bullfrog (Rana catesbeiana) was evaluated by the North Carolina State University College of Veterinary Medicine's Turtle Rescue Team following vehicular trauma. A fracture of the left femur was suspected on examination and palpation of the hind limbs, but no other injuries or abnormalities were detected. While the bullfrog was sedated, whole-body radiographic views were obtained, which revealed a closed midshaft comminuted fracture of the left femur. The fracture was repaired by use of an internal fixation technique that included Kirschner wires, a positive-profile pin secured along the femur with encircling sutures, and polymethylmethacrylate molded around the entire apparatus. There were no major complications during the postoperative rehabilitation period. One year after surgery, radiography revealed complete fracture healing and the bullfrog was released back into the wild. Presently, there are no widely accepted methods for fracture fixation in amphibians. Factors associated with their aquatic environment and lengthy fracture healing time must be addressed when planning fracture fixation strategies. In the bullfrog of this report, the applied internal fixation method provided effective long-term stabilization of the femur, allowed for normal movement, and enabled the bullfrog to be housed in an aquatic environment immediately after surgery.


Tear exchange under hydrogel contact lenses: methodological considerations.
To characterize hydrogel lens tear exchange and to apply an optimized method to compare tear exchange of a marketed hydrogel lens to that measured with a prototype silicone hydrogel lens. Fluorophotometry and a nonpenetrating tracer (70-kDa FITC-dextran) were used with a single extended-wear soft contact lens (EWSCL) material on 11 subjects to characterize tear-exchange kinetics. Twenty to 30 measurements were obtained over a 30-minute period to allow accurate modeling and estimation of the several tear-exchange parameters. Calculated values included tear-replenishment rate (TRR), elimination rate (ER), and the time for 95% of the signal to be eliminated (T(95)). Major experiments were (1) comparison of ER under controlled and physiological conditions, (2) comparison of right and left eyes, (3) repeatability of ER and T(95) on five occasions, and (4) comparison of a marketed lens (oxygen permeability [Dk] 28 x 10(-9) [cm/sec][ml O(2)/ml mm Hg]) to a prototype silicone hydrogel lens (Dk 140 x 10(-9) [cm/sec][ml O(2)/ml mm Hg]). Tracer elimination behind a hydrogel contact lens (CL), up to 30 minutes after insertion, was optimally described by double-exponential kinetics. Physiological ER (5-30 minutes after CL insertion) was optimally described by single-exponential kinetics. Overall, physiological ER was 8.8% +/- 3.8% per minute, and T(95) was 31.0 +/- 16.1 minutes (n = 76 and 72 determinations, respectively). Differences between right and left eyes in ER and T(95) were not significant at the 0.05 level. No difference in ER or T(95) was found between habitual and controlled blinking. Mean TRR was 0.67% +/- 0.26% per blink (n = 11 determinations). No differences were shown between ER or T(95) measurements over time. A prototype highly oxygen-permeable silicone hydrogel lens showed higher ER than did a marketed hydrogel lens (P < 0.01). Estimates of postlens tear exchange using a slit lamp fluorophotometer are similar to previously reported rates using similar fluorophotometric techniques. Fluorescent decay behind a hydrogel lens is most precisely described using a double-exponential curve equation and tear exchange may be described using ER, TRR and T(95), although the T(95) may be the least reliable of these measures. The technique appears capable of discriminating between lens types.


New neural-network-based method to infer total ozone column amounts and cloud effects from multi-channel, moderate bandwidth filter instruments.
A new method is presented based on a radial basis function neural network (RBF-NN) to analyze data obtained by ultraviolet (UV) irradiance instruments. Application of the RBF-NN method to about three years of data obtained by a NILU-UV device, which is a multi-channel, moderate bandwidth filter instrument, revealed that compared to the traditional Look-up table (LUT) method, the RBF-NN method yielded better agreement with a 1% decrease in relative difference and an increase of 0.03 in correlation with total ozone column (TOC) values obtained from the Ozone Monitoring Instrument (OMI). Furthermore, the RBF-NN method retrieved more valid results (daily average values within a meaningful range (200-500 DU)) than the LUT method. Compared with RBF-NN retrievals, TOC values obtained from the OMI are underestimated under cloudy conditions. This finding agrees with conclusions reached by Anton and Loyola (2011).


Evaluation of a whole-genome amplification method based on adaptor-ligation PCR of randomly sheared genomic DNA.
High-throughput genetic studies often require large quantities of DNA for a variety of analyses. Developing and assessing a whole-genome amplification method is thus important, especially with the current desire for large-scale genotyping in previously collected samples for which limited DNA is available. The method we have developed, called PRSG, is based on an adaptor-ligation-mediated PCR of randomly sheared genomic DNA. An unbiased representation was evaluated by performing PCR on 2,607 exons of 367 genes, which are randomly distributed throughout the genome, on PRSG products of hundreds of individuals. An infrequent loss (<1%) of the exon sequence on the PRSG products was found. Out of 307 microsatellites on various chromosomes, 258 (84%) were amplified in both the PRSG product and an original DNA, whereas 49 (16%) microsatellites were lost only in the PRSG product. Array CGH analysis of 287 loci for measuring the relative gene copy number demonstrated that a low bias was detected. Moreover, this method was validated on 100-1,000 laser-captured cells from paraffin-embedded tissues. These data show that PRSG can provide a sufficient amount of genomic sequence for a variety of genetic analyses as well as for long-term storage for future work.


Complete genome sequence of the Variibacter gotjawalensis GJW-30(T) from soil of lava forest, Gotjawal.
Variibacter gotjawalensis GJW-30(T) is a gram-negative, strictly aerobic bacterium to form pleomorphic. Here we present the 4.5-Mb genome sequence of the type strain of V. gotjawalensis GJW-30(T), which consists a chromosome for the total 4,586,237bp with a G+C content of 62.2mol%. This is the first report of the full genome sequence of a species of the novel genus Variibacter isolated from Gotjawal, a unique area in Jeju, Republic of Korea.