Triggers of subarachnoid hemorrhage: role of physical exertion, smoking, and alcohol in the Australasian Cooperative Research on Subarachnoid Hemorrhage Study (ACROSS).
Unaccustomed strenuous physical exertion can trigger myocardial infarction, but little is known about the mechanisms precipitating subarachnoid hemorrhage (SAH). We identified all cases of first-ever SAH among the combined populations (2.8 million) of 4 urban centers in Australia and New Zealand. Information on the type, time, and intensity of exposures in the 26 hours before the onset of SAH was ascertained by structured interviews. We used the case-crossover technique to assess the risk of SAH associated with transient exposures of moderate to extreme physical exertion, heavy cigarette smoking, and binge alcohol consumption. We registered 432 first-ever cases of SAH (62% women; mean age, 56.5 years). A definite time of onset of SAH was established for 393 patients (91%), and information on the levels of physical activity in the preceding 26 hours was obtained in 338 (78%). Of these patients, 19% engaged in moderate to extreme exertion (>or=5 metabolic equivalents) in the 2 hours before SAH, which was associated with a tripling in the risk of SAH (odds ratio [OR], 2.7; 95% CI, 1.6 to 4.6). There was no evidence of any association between heavy cigarette smoking or binge drinking and risk of SAH in the subsequent 2 hours (OR, 1.1; 95% CI, 0.4 to 3.7; and OR, 0.41; 95% CI, - infinity to 5.3). Habitual exercise did not appear to alter the risk of SAH associated with moderate to extreme exertion. Moderate to extreme physical exertion tripled the risk of SAH, but there was no association between transient heavy smoking or binge drinking and risk of SAH. These data suggest that heavy physical activity may trigger SAH.


Overexpression of MazFsa in Staphylococcus aureus induces bacteriostasis by selectively targeting mRNAs for cleavage.
The role of chromosomally encoded toxin-antitoxin (TA) loci in bacterial physiology has been under debate, with the toxin proposed as either an inducer of bacteriostasis or a mediator of programmed cell death (PCD). We report here that ectopic expression of MazF(Sa), a toxin of the TA module from Staphylococcus aureus, led to a rapid decrease in CFU counts but most cells remained viable as determined by differential Syto 9 and propidium iodide staining after MazF(Sa) induction. This finding suggested that the toxin MazF(Sa) induced cell stasis rather than cell death. We also showed that MazF(Sa) selectively cleaves cellular mRNAs in vivo, avoiding "important" transcripts such as recA, gyrB, and sarA mRNAs in MazF(Sa)-induced cells, while these three mRNAs can be cleaved in vitro. The results of Northwestern blotting showed that both sarA and recA mRNAs bind strongly to a putative RNA-binding protein. These data suggest that S. aureus likely undergoes stasis by protecting selective mRNA with RNA-binding proteins upon the expression of MazF(Sa) in vivo.


Myocardial scar visualized by cardiovascular magnetic resonance imaging predicts major adverse events in patients with hypertrophic cardiomyopathy.
We sought to establish the prognostic value of a comprehensive cardiovascular magnetic resonance (CMR) examination in risk stratification of hypertrophic cardiomyopathy (HCM) patients. With annual mortality rates ranging between 1% and 5%, depending on patient selection, a small but significant number of HCM patients are at risk for an adverse event. Therefore, the identification of and prophylactic therapy (i.e., defibrillator placement) in patients with HCM who are at risk of dying are imperative. Two-hundred forty-three consecutive patients with HCM were prospectively enrolled. All patients underwent initial CMR, and 220 were available for clinical follow-up. The mean follow-up time was 1,090 days after CMR. End points were all-cause and cardiac mortality. During follow-up 20 of the 220 patients died, and 2 patients survived sudden cardiac death due to adequate implantable cardioverter-defibrillator discharge. Most events (n = 16) occurred for cardiac reasons; the remaining 6 events were related to cancer and accidents. Our data indicate that the presence of scar visualized by CMR yields an odds ratio of 5.47 for all-cause mortality and of 8.01 for cardiac mortality. This might be superior to classic clinical risk factors, because in our dataset the presence of 2 risk factors yields an odds ratio of 3.86 for all-cause and of 2.20 for cardiac mortality, respectively. Multivariable analysis also revealed the presence of late gadolinium enhancement as a good independent predictor of death in HCM patients. Among our population of largely low or asymptomatic HCM patients, the presence of scar indicated by CMR is a good independent predictor of all-cause and cardiac mortality.


T4-like Phages Reveal the Potential Role of Viruses in Soil Organic Matter Mineralization.
Viruses are the most abundant biological entities in the world, but their ecological functions in soil are virtually unknown. We hypothesized that greater abundance of T4-like phages will increase bacterial death and thereby suppress soil organic carbon (SOC) mineralization. A range of phage and bacterial abundances were established in sterilized soil by reinoculation with 10-3 and 10-6 dilutions of suspensions of unsterilized soil. The total and viable 16S rRNA gene abundance (a universal marker for bacteria) was measured by qPCR to determine bacterial abundance, with propidium monoazide (PMA) preapplication to eliminate DNA from non-viable cells. Abundance of the g23 marker gene was used to quantify T4-like phages. A close negative correlation between g23 abundance and viable 16S rRNA gene abundance was observed. High abundance of g23 led to lower viable ratios for bacteria, which suggested that phages drove microbial necromass production. The CO2 efflux from soil increased with bacterial abundance but decreased with higher abundance of T4-like phages. Elimination of extracellular DNA by PMA strengthened the relationship between CO2 efflux and bacterial abundance, suggesting that SOC mineralization by bacteria is strongly reduced by the T4-like phages. A random forest model revealed that abundance of T4-like phages and the abundance ratio of T4-like phages to bacteria are better predictors of SOC mineralization (measured as CO2 efflux) than bacterial abundance. Our study provides experimental evidence of phages' role in organic matter turnover in soil: they can retard SOC decomposition but accelerate bacterial turnover.


Cuing both positive and negative episodic foresight reduces delay discounting but does not affect risk-taking.
Humans frequently create mental models of the future, allowing outcomes to be inferred in advance of their occurrence. Recent evidence suggests that imagining positive future events reduces delay discounting (the devaluation of reward with time until its receipt), while imagining negative future events may increase it. Here, using a sample of 297 participants, we experimentally assess the effects of cued episodic simulation of positive and negative future scenarios on decision-making in the context of both delay discounting (monetary choice questionnaire) and risk-taking (balloon-analogue risk task). Participants discounted the future less when cued to imagine positive and negative future scenarios than they did when cued to engage in control neutral imagery. There were no effects of experimental condition on risk-taking. Thus, although these results replicate previous findings suggesting episodic future simulation can reduce delay discounting, they indicate that this effect is not dependent on the valence of the thoughts, and does not generalise to all other forms of "impulsive" decision-making. We discuss various interpretations of these results, and suggest avenues for further research on the role of prospection in decision-making.


Improving the evaluation and diagnosis of clinically significant prostate cancer.
This chapter describes important changes in the clinical practice and pathological interpretation of prostate cancer (PCa) that stratify PCa into insignificant and significant disease to allow for better patient management and treatment decisions. Among the most important changes over the last half a decade has been the change in PCa grading. A new grading system has been developed that is more patient-centric with more accurate stratification according to risk of disease progression. An additional advance is the recent recommendation to report percentage of pattern 4 in Gleason score 7 cancer that may help identify a subset of these men who have relatively insignificant prostate cancer. The definition of significant PCa at prostatectomy has also been extended from a dichotomized classification into a more tiered nuanced approach. Factors involved in classification of significant cancer at prostatectomy include grade, stage along with classification of extraprostatic extension into focal and nonfocal, and tumor volume. Of these variables, tumor volume appears to be least critical as an independent variable. Promising genetic discoveries may also help predict significant cancer, especially in the small subset of lower-grade cancers. Significant PCa is a spectrum of findings bearing different clinical significance with some nuanced definitions at biopsy and prostatectomy.


Heartbeat perception in panic disorder: a reanalysis.
This article describes a reanalysis of seven studies on heart beat perception (HBP) in panic disorder. The pooled sample had 709 participants from eight diagnostic categories. Accurate HBP was uncommon, but more prevalent among panic disorder patients than among healthy controls, depressed patients, patients with palpitations and individuals with infrequent panic attacks. No differences were found between panic disorder patients and patients with other anxiety disorders. Accurate perceivers had higher anxiety sensitivity scores than inaccurate perceivers. The data remain inconclusive as to whether perceived heart rate is correlated with anxiety in inaccurate perceivers. Physical exercise, distraction, variations in instructions and treatment each influenced HBP. However, the influence was different than previously thought. Finally, it is suggested that HBP may be understood in terms of schema-guided information processing.


How do cardiovascular diseases harm labor force participation? Evidence of nationally representative survey data from Japan, a super-aged society.
To evaluate how cardiovascular diseases harm labor force participation (LFP) among the Japanese population and verify the validity of plasma biomarkers as instrumental variables of cardiovascular diseases after adjusting for a broad set of confounders including dietary intake. Using nationally representative repeated cross-sectional surveys in Japan, the Comprehensive Survey of Living Conditions and National Health and Nutrition Survey, with plasma biomarkers as instrumental variables for quasi-randomization. Onset of cardiovascular diseases in those receiving regular treatment for hypertension, intracerebral hemorrhage, intracerebral infarction, angina pectoris, myocardial infarction, or other types of cardiovascular diseases. A total of 65,615 persons aged ≥ 20 years (35,037 women and 30,578 men) who completed a survey conducted every three years from 1995 through 2013. Respondent employment and weekly working hours during each survey year. Cardiovascular diseases significantly and remarkably reduced the probability of working by 15.4% (95% CI: -30.6% to -0.2%). The reduction in working probability was detected for women only. Respondents aged ≥ 40 years were less likely to work once diagnosed and the reduction was enlarged for those aged ≥ 65 years, while those aged < 40 years appeared to be unaffected. Probability of engaging in manual labor significantly decreased once diagnosed; however, no impact was found for cognitive occupations. Among employed respondents, the adverse effects of cardiovascular diseases decreased working hours by five hours per week. Validity of the biomarker instrumental variables was generally verified. A vicious circle is suggested between LFP and unfavorable health. However, the effects vary across age, sex, and occupation type, even after adjusting for causal effects, which could cause a downward bias in LFP impact. cardiovascular disease, labor force participation, instrumental variable method as quasi-randomization, plasma biomarker, Comprehensive Survey of Living Conditions, National Health and Nutrition Survey.


A novel design process for selection of attributes for inclusion in discrete choice experiments: case study exploring variation in clinical decision-making about thrombolysis in the treatment of acute ischaemic stroke.
A discrete choice experiment (DCE) is a method used to elicit participants' preferences and the relative importance of different attributes and levels within a decision-making process. DCEs have become popular in healthcare; however, approaches to identify the attributes/levels influencing a decision of interest and to selection methods for their inclusion in a DCE are under-reported. Our objectives were: to explore the development process used to select/present attributes/levels from the identified range that may be influential; to describe a systematic and rigorous development process for design of a DCE in the context of thrombolytic therapy for acute stroke; and, to discuss the advantages of our five-stage approach to enhance current guidance for developing DCEs. A five-stage DCE development process was undertaken. Methods employed included literature review, qualitative analysis of interview and ethnographic data, expert panel discussions, a quantitative structured prioritisation (ranking) exercise and pilot testing of the DCE using a 'think aloud' approach. The five-stage process reported helped to reduce the list of 22 initial patient-related factors to a final set of nine variable factors and six fixed factors for inclusion in a testable DCE using a vignette model of presentation. In order for the data and conclusions generated by DCEs to be deemed valid, it is crucial that the methods of design and development are documented and reported. This paper has detailed a rigorous and systematic approach to DCE development which may be useful to researchers seeking to establish methods for reducing and prioritising attributes for inclusion in future DCEs.


Clinical Manifestation, Imaging Features and Treatment Follow-up of 29 Cases with Hepatic Tubercolosis.
To understand the clinical and imaging manifestations and the treatment and follow-up of hepatic tuberculosis (HTB), we retrospectively analysed the clinical and imaging data of 29 patients with HTB who had been diagnosed clinically or by biopsy, and the clinical and imaging data had been summarised. Patient characteristics were followed up after anti-TB drug treatment. The median age of the 29 patients with HTB was 37 years, and most were male (58.6%). The patient's symptoms included fever (48.2%), respiratory symptoms (27.5%), abdominal pain (24.1%), and abdominal distension (10.3%). Elevated erythrocyte sedimentation rate (79.3%), elevated serum C-reactive protein (75.8%) and hypoalbuminemia (62.0%) were common features. Three patients were serologically positive for acquired human immunodeficiency syndrome, and two were serologically positive for hepatitis B surface antigen with normal tumour markers. The 29 patients with HTB included 17 with serous HTB, 9 with parenchymal HTB (8 with parenchymal nodular HTB and 1 with parenchymal miliary HTB), 1 with intrahepatic abscess type HTB, and 2 with hilar HTB. Approximately 86% of the patients also had pulmonary TB. Most of the serous HTB patients also had tuberculous peritonitis. Enhanced computerized tomography scans of the serous and parenchymal HTB cases showed the progressive development of lesions. Abnormal blood perfusion was observed in the hepatic artery, and the clearest evidence of TB was observed in the hepatic portal vein. Magnetic resonance imaging indicated that the lesions returned a high signal in the diffusion-weighted imaging sequence. However, the lesions' apparent diffusion coefficient values reflected high signals. The Xpert MTB/RIF test detected Mycobacterium TB complex in the liver biopsy fluid from 10 patients. Regarding histopathology, one patient showed granulomatous inflammation, and one patient's acid-fast bacillus (AFB) stain was positive. The treatment of two patients was stopped due to their adverse reactions to the drugs and the risk of creating drug-resistant TB. The remaining patients received anti-TB treatment, but one subsequently died, and two were unavailable for follow-up. The clinical symptoms of HTB are difficult to detect, and it has diverse manifestations by imaging, with no obvious specificity in terms of pathological results. Therefore, follow-up of liver lesions for checking anti-TB therapy is another method for diagnosing HTB. In addition, early active anti-TB treatment can achieve good curative results.


Temporal Segregation between Dung-Inhabiting Beetle and Fly Species.
The coexistence of ecologically similar species (i.e. species utilizing the same resource) is a major topic in ecology. Communities are assembled either through the biotic interactions of ecologically similar species, e.g. competition, or by the abiotic separation of species along gradients of environmental conditions. Here, we investigated the temporal segregation, succession and seasonality of dung-inhabiting Coleoptera and Diptera that utilize an identical resource in exactly the same way. The data were collected from two temperate pastures, one in the United Kingdom and the second in the Czech Republic. There was no evident temporal separation between ecologically similar coleopterous or dipterous taxa during succession. In contrast, these two orders were almost perfectly separated seasonally in both combined and site-specific datasets. Flies were most abundant in the summer, and beetles were more abundant in the spring and autumn. Ecologically similar beetles and flies also displayed seasonal separation in both combined and site-specific data. Analyses within site-specific data sets revealed such a separation at both the order and species level. Season is therefore the main temporal axis separating ecologically similar species of dung-inhabiting insects in temperate habitats, while succession aggregates species that may have similar environmental tolerances (to e.g. dung moisture). This separation between ecologically similar taxa of beetles and flies may be attributable to either competition-based niche separation or to temperature tolerance-based habitat filtering, since flies have peak activity in warmer months while beetles have peak activity in cooler months.


Investigating online psychological treatment for adolescents with a visible difference in the Dutch YP Face IT study: protocol of a randomised controlled trial.
This paper outlines the study protocol for the Dutch Young People (YP) Face IT Study. Adolescents with a visible difference (ie, disfigurement) often experience challenging social situations such as being stared at, receiving unwanted questions or being teased. As a consequence, some of these adolescents experience adverse psychosocial outcomes and appearance-related distress. To address this appearance-related distress, an online psychotherapeutic intervention, YP Face IT, has been developed. YP Face IT uses social interaction skills training and cognitive-behavioural therapy. The Dutch YP Face IT Study tests whether this intervention is effective in reducing social anxiety and improving body esteem. Participants are adolescents aged 12-18 with a visible difference and experiencing appearance-related distress. In this two-armed randomised controlled trial, 224 adolescents will be randomised to care as usual or YP Face IT. Adolescents will complete questionnaires at baseline, at 13 weeks and at 25 weeks. Primary outcomes are differences in social anxiety and body esteem between YP Face IT and care as usual. Secondary outcomes are differences in aspects of self-worth, perceived stigmatisation, health-related quality of life, life engagement, appearance-related distress and depressive symptoms between the two groups. Research ethics approval was obtained from the medical ethics review committee in Rotterdam (reference number MEC-2018-052/NL63955.078.18). Findings will be disseminated through academic peer-reviewed publications, conferences and newsletters to patient associations and participants of the study. The Netherlands Trial Register (NL7626).


Neuroendoscopic Technique for Recurrent Chronic Subdural Hematoma with Small Craniotomy.
To present a case series describing an endoscopic technique with a small craniotomy for recurrent chronic subdural hematoma (rCSDH) treatment. A total of 17 patients with rCSDH underwent neuroendoscopic hematoma removal with a small craniotomy under local or general anesthesia. The skin incision of the initial surgery on the convexity of the skull was extended, and a burr hole was created for a small craniotomy. After the removal of the outer membrane and hematoma through a small craniotomy, the hematoma was evacuated with a suction tube using the rigid endoscope. The entire hematoma cavity circumference was irrigated, while septations and trabeculae in the hematoma were cut. After hematoma evacuation, the inner membrane was incised and removed to allow brain expansion. Postoperative follow-up was performed for at least 6 months. The regrowth rate of rCSDH after the neuroendoscopy was 5.9%. One patient with recurrent chronic subdural hematoma regrowth required neuroendoscopy again, but no re-recurrence was observed for the next 6 months. All cases were successfully managed using this technique and the postoperative seizure rate was 23.5%. This neuroendoscopic technique with a small craniotomy could be useful for recurrent chronic subdural hematoma because the hematoma and septations can be visualized and evacuated along the entire circumference of the hematoma cavity, and the inner membrane can be torn to allow brain expansion.


Serum exosomal-annexin A2 is associated with African-American triple-negative breast cancer and promotes angiogenesis.
Limited information is available on biomarker(s) for triple-negative breast cancer (TNBC) that can address the higher incidence and aggressiveness of TNBC in African-American (AA) women. Our previous studies have demonstrated annexin A2 (AnxA2) association with exosomes which promotes angiogenesis and metastasis. Therefore, our goal was to examine the expression and function of exosomal-annexin A2 (exo-AnxA2) derived from the serum samples of breast cancer patients. The expression of serum exo-AnxA2 and its association with clinicopathological features of the breast cancer patients were determined. The role of serum exo-AnxA2 to promote angiogenesis was determined by an in vivo Matrigel plug assay. Our results show that the expression of serum exo-AnxA2 in breast cancer patients (n = 169; 83.33 ± 2.040 ng/mL, P < 0.0001) is high compared to non-cancer females (n = 68; 34.21 ± 2.238 ng/mL). High expression of exo-AnxA2 levels in breast cancer was significantly associated with tumor grade (P < 0.0001), poor overall survival (hazard ratio (HR) 2.802; 95% confidence intervals (CI) = 1.030-7.620; P = 0.0353), and poor disease-free survival (HR 7.934; 95% CI = 1.778-35.398; P = 0.0301). The expression of serum exo-AnxA2 levels was significantly elevated in TNBC (n = 68; 109.1 ± 2.905 ng/mL; P < 0.0001) in comparison to ER+ (n = 50; 57.35 ± 1.545 ng/mL), HER2+ (n = 59; 78.25 ± 1.146 ng/mL), and non-cancer females (n = 68; 34.21 ± 2.238 ng/mL). Exo-AnxA2 showed diagnostic values with a maximum AUC as 1.000 for TNBC, 0.8304 for ER+, and 0.9958 for HER2+ compared to non-cancer females. The expression of serum exo-AnxA2 was significantly elevated in AA women with TNBC (n = 29; 118.9 ± 4.086 ng/mL, P < 0.0001) in comparison to Caucasian-American TNBC (n = 27; 97.60 ± 3.298 ng/mL) patients. Our in vivo results suggest a role of serum exo-AnxA2 in angiogenesis and its association with aggressiveness of TNBC in AA women. Our results demonstrated that the expression of serum exo-AnxA2 is high in AA women with TNBC and promotes angiogenesis. These findings suggest that exo-AnxA2 holds promise as a potential prognosticator of TNBC and may lead to an effective therapeutic option.


The collateral effects of joint attention training on social initiations, positive affect, imitation, and spontaneous speech for young children with autism.
Joint attention may be a core deficit in autism which underlies the abnormal development of later emerging social-communication behaviors. Given this theory, researchers have suggested that teaching young children with autism to engage in joint attention may lead to collateral increases in other non-targeted social-communication behaviors. In this study, children with autism participated in a 10-week joint attention training program and collateral changes in non-targeted behaviors were assessed. Following participation in the intervention, positive collateral changes were observed in social initiations, positive affect, imitation, play, and spontaneous speech. Results support the hypothesis that teaching joint attention skills leads to improvement in a variety of related skills and have implications for the treatment of young children with autism.


COMETARY SCIENCE. Thermal and mechanical properties of the near-surface layers of comet 67P/Churyumov-Gerasimenko.
Thermal and mechanical material properties determine comet evolution and even solar system formation because comets are considered remnant volatile-rich planetesimals. Using data from the Multipurpose Sensors for Surface and Sub-Surface Science (MUPUS) instrument package gathered at the Philae landing site Abydos on comet 67P/Churyumov-Gerasimenko, we found the diurnal temperature to vary between 90 and 130 K. The surface emissivity was 0.97, and the local thermal inertia was 85 ± 35 J m(-2) K(-1)s(-1/2). The MUPUS thermal probe did not fully penetrate the near-surface layers, suggesting a local resistance of the ground to penetration of >4 megapascals, equivalent to >2 megapascal uniaxial compressive strength. A sintered near-surface microporous dust-ice layer with a porosity of 30 to 65% is consistent with the data.


A Mobile Phone Based Method to Assess Energy and Food Intake in Young Children: A Validation Study against the Doubly Labelled Water Method and 24 h Dietary Recalls.
Mobile phones are becoming important instruments for assessing diet and energy intake. We developed the Tool for Energy Balance in Children (TECH), which uses a mobile phone to assess energy and food intake in pre-school children. The aims of this study were: (a) to compare energy intake (EI) using TECH with total energy expenditure (TEE) measured via doubly labelled water (DLW); and (b) to compare intakes of fruits, vegetables, fruit juice, sweetened beverages, candy, ice cream, and bakery products using TECH with intakes acquired by 24 h dietary recalls. Participants were 39 healthy, Swedish children (5.5 ± 0.5 years) within the ongoing Mobile-based Intervention Intended to Stop Obesity in Preschoolers (MINISTOP) obesity prevention trial. Energy and food intakes were assessed during four days using TECH and 24 h telephone dietary recalls. Mean EI (TECH) was not statistically different from TEE (DLW) (5820 ± 820 kJ/24 h and 6040 ± 680 kJ/24 h, respectively). No significant differences in the average food intakes using TECH and 24 h dietary recalls were found. All food intakes were correlated between TECH and the 24 h dietary recalls (ρ = 0.665-0.896, p < 0.001). In conclusion, TECH accurately estimated the average intakes of energy and selected foods and thus has the potential to be a useful tool for dietary studies in pre-school children, for example obesity prevention trials.


Systematic review of clinical features of suspected prostate cancer in primary care.
To systematically review the literature and provide an update and integration of existing peer-reviewed guidelines with recent systematic reviews and with primary studies related to the early recognition and management of prostate cancer in primary care. We searched MEDLINE and EMBASE for relevant articles. The quality of the evidence to support existing guideline recommendations and the consistency of recommendations with updated evidence were assessed. Applicability in a Canadian primary care setting was also evaluated. All studies conducted in the primary care setting that provided information on clinical features predictive of prostate cancer were included. Also, studies that assessed the accuracy of nomograms to predict prostate cancer were reviewed. The findings suggest that lower urinary tract symptoms are not highly predictive of prostate cancer. However, evidence suggests that FPs might be good at discriminating between patients with and without prostate cancer using digital rectal examination and prostate-specific antigen testing. Nomograms might also be useful in assessing patients for aggressive prostate cancers. The results of this review can be used to inform recommendations for referral for suspected prostate cancer in the primary care setting. They could also inform development of prostate cancer diagnostic assessment programs.


Trappin-2 promotes early clearance of Pseudomonas aeruginosa through CD14-dependent macrophage activation and neutrophil recruitment.
Microaspiration of Pseudomonas aeruginosa contributes to the pathogenesis of nosocomial pneumonia. Trappin-2 is a host defense peptide that assists with the clearance of P. aeruginosa through undefined mechanisms. A model of macrophage interactions with replicating P. aeruginosa (strain PA01) in serum-free conditions was developed, and the influence of subantimicrobial concentrations of trappin-2 was subsequently studied. PA01 that was pre-incubated with trappin-2 (at concentrations that have no direct antimicrobial effects), but not control PA01, was cleared by alveolar and bone marrow-derived macrophages. However, trappin-2-enhanced clearance of PA01 was completely abrogated by CD14- null macrophages. Fluorescence microscopy demonstrated the presence of trappin-2 on the bacterial cell surface of trappin-2-treated PA01. In a murine model of early lung infection, trappin-2-treated PA01 was cleared more efficiently than control PA01 2 hours of intratracheal instillation. Furthermore, trappin-2-treated PA01 up-regulated the murine chemokine CXCL1/KC after 2 hours with a corresponding increase in neutrophil recruitment 1 hour later. These in vivo trappin-2-treated PA01 effects were absent in CD14-deficient mice. Trappin-2 appears to opsonize P. aeruginosa for more efficient, CD14-dependent clearance by macrophages and contributes to the induction of chemokines that promote neutrophil recruitment. Trappin-2 may therefore play an important role in innate recognition and clearance of pathogens during the very earliest stages of pulmonary infection.


Store-Operated Ca2+ Channels: Mechanism, Function, Pharmacology, and Therapeutic Targets.
Calcium (Ca2+) release-activated Ca2+ (CRAC) channels are a major route for Ca2+ entry in eukaryotic cells. These channels are store operated, opening when the endoplasmic reticulum (ER) is depleted of Ca2+, and are composed of the ER Ca2+ sensor protein STIM and the pore-forming plasma membrane subunit Orai. Recent years have heralded major strides in our understanding of the structure, gating, and function of the channels. Loss-of-function and gain-of-function mutants combined with RNAi knockdown strategies have revealed important roles for the channel in numerous human diseases, making the channel a clinically relevant target. Drugs targeting the channels generally lack specificity or exhibit poor efficacy in animal models. However, the landscape is changing, and CRAC channel blockers are now entering clinical trials. Here, we describe the key molecular and biological features of CRAC channels, consider various diseases associated with aberrant channel activity, and discuss targeting of the channels from a therapeutic perspective.


Emergence of a Novel Chimeric Gene Underlying Grain Number in Rice.
Grain number is an important factor in determining grain production of rice (Oryza sativa L.). The molecular genetic basis for grain number is complex. Discovering new genes involved in regulating rice grain number increases our knowledge regarding its molecular mechanisms and aids breeding programs. Here, we identified GRAINS NUMBER 2 (GN2), a novel gene that is responsible for rice grain number, from "Yuanjiang" common wild rice (O. rufipogon Griff.). Transgenic plants overexpressing GN2 showed less grain number, reduced plant height, and later heading date than control plants. Interestingly, GN2 arose through the insertion of a 1094-bp sequence from LOC_Os02g45150 into the third exon of LOC_Os02g56630, and the inserted sequence recruited its nearby sequence to generate the chimeric GN2 The gene structure and expression pattern of GN2 were distinct from those of LOC_Os02g45150 and LOC_Os02g56630 Sequence analysis showed that GN2 may be generated in the natural population of Yuanjiang common wild rice. In this study, we identified a novel functional chimeric gene and also provided information regarding the molecular mechanisms regulating rice grain number.


The p75 neurotrophin receptor in AgRP neurons is necessary for homeostatic feeding and food anticipation.
Networks of neurons control feeding and activity patterns by integrating internal metabolic signals of energy balance with external environmental cues such as time-of-day. Proper circadian alignment of feeding behavior is necessary to prevent metabolic disease, and thus it is imperative that molecular players that maintain neuronal coordination of energy homeostasis are identified. Here, we demonstrate that mice lacking the p75 neurotrophin receptor, p75NTR, decrease their feeding and food anticipatory behavior (FAA) in response to daytime, but not nighttime, restricted feeding. These effects lead to increased weight loss, but do not require p75NTR during development. Instead, p75NTR is required for fasting-induced activation of neurons within the arcuate hypothalamus. Indeed, p75NTR specifically in AgRP neurons is required for FAA in response to daytime restricted feeding. These findings establish p75NTR as a novel regulator gating behavioral response to food scarcity and time-of-day dependence of circadian food anticipation.


Physiological advantage of the mechanism of the tryptophan synthetase reaction.
Indole-accumulating and indole-utilizing tryptophan synthetase mutants of Saccharomyces cerevisiae were found to form complementing diploids with unusual growth properties. These strains grew at nearly the wild-type exponential rate, but only after an abnormally prolonged lag period of a duration that depended on the initial cell density. The exaggerated lag was reduced by adding low concentrations of indole in the medium. The growth properties were interpreted to be a consequence of the inability of these strains to synthesize tryptophan by the normal mechanism.


Current advances in sepsis and septic shock with particular emphasis on the role of insulin.
Sepsis and septic shock account for substantial morbidity and mortality in the intensive care units. NF-kappaB activation, and elevated concentrations of macrophage migration inhibitory factor (MIF), tumor necrosis factor-a (TNF-alpha), interleukin-1 (IL-1), IL-6, free radicals, inducible nitric oxide (iNO), and stress hyperglycemia are some of the factors that induce systemic inflammatory response and myocardial depression seen in sepsis. Conversely, adenosine, activated protein C, oxidized phospholipids, w-3 fatty acids, and insulin have beneficial effects in sepsis and septic shock. These molecules and in particular insulin have the ability to suppress synthesis of MIF, TNF-alpha, IL-1, IL-6, and free radicals, enhance endothelial NO production, and enhance the production of anti-inflammatory cytokines IL-10, and IL-4. In addition, insulin corrects stress hyperglycemia and improves myocardial function. Thus insulin, adenosine, activated protein C, oxidized phospholipids, and w-fatty acids show anti-inflammatory actions and explain why and how they are useful in sepsis and septic shock and possibly, other inflammatory conditions. Hence, their combined use may be of significant benefit in sepsis and septic shock.


Contribution of N-methyl-D-aspartate receptors to attention and episodic spatial memory during senescence.
A decrease in N-methyl-D-aspartate receptor (NMDAR) function is associated with age-related cognitive impairments. However, NMDAR antagonists are prescribed for cognitive decline associated with age-related neurodegenerative disease, raising questions as to the role of NMDAR activity in cognitive function during aging. The current studies examined effects of NMDAR blockade on cognitive task that are sensitive to aging. Young and middle-age rats were trained on the five-choice serial reaction time task (5-CSRTT) and challenged with MK-801 (0.025, 0.05, and 0.1mg/kg or vehicle). Attention deficits were apparent in middle-age and performance of young and middle-age rats was enhanced for low doses of MK-801 (0.025 and 0.05). The beneficial effects on attention were reversed by the highest dose of MK-801. Older animals exhibited a delay-dependent impairment of episodic spatial memory examined on a delayed-matching to place water maze task. Similarly, a low dose of MK-801 (0.05mg/kg) impaired performance with increasing delay and aged animals were more susceptible to disruption by NMDAR blockade. Despite MK-801 impairment of episodic spatial memory, MK-801 had minimal effects on spatial reference memory. Our results confirm that NMDARs contribute to rapidly acquired and flexible spatial memory and support the idea that a decline in NMDAR function contributes to the age-related impairments in cognition.


A window of opportunity: declining rates of hepatitis B virus infection among injection drug users in Rio de Janeiro, and prospects for targeted hepatitis B vaccination.
To measure hepatitis B virus (HBV) infection rates among injection drug users in Rio de Janeiro, Brazil, and to report their knowledge of and attitudes toward hepatitis and HBV vaccination. 609 injection drug users recruited in Rio de Janeiro between 1999 and 2001 answered a questionnaire and were tested for hepatitis B and other blood-borne infections. Questions covered sociodemographic information, alcohol and illicit drug consumption, drug injection and sexual practices, medical history, and knowledge about HIV, AIDS and viral hepatitis. The prevalence of HBV infection was 27.1%, with 3.4% of the sample positive for HbsAg (active infection) and 0.8% positive for anti-HBs (indicating previous HBV vaccination). Most interviewees (81.3%) were aware of at least one form of viral hepatitis and received information from many different sources. In agreement with laboratory findings, 96.7% of the interviewees stated they had never been vaccinated against hepatitis B, but almost all unvaccinated interviewees (97.8%) said they would volunteer to be vaccinated if HBV vaccination were available. Few of the injection drug users surveyed had ever been vaccinated against HBV. Although most were aware of the risks posed by viral hepatitis, this awareness seldom translated into consistent behavioral change. The participants' willingness to be vaccinated against HBV suggests that the implementation of vaccination for this population may help decrease rates of hepatitis B infection.


The programmed death-1 ligand 1:B7-1 pathway restrains diabetogenic effector T cells in vivo.
Programmed death-1 ligand 1 (PD-L1) is a coinhibitory molecule that negatively regulates multiple tolerance checkpoints. In the NOD mouse model, PD-L1 regulates the development of diabetes. PD-L1 has two binding partners, programmed death-1 and B7-1, but the significance of the PD-L1:B7-1 interaction in regulating self-reactive T cell responses is not yet clear. To investigate this issue in NOD mice, we have compared the effects of two anti-PD-L1 Abs that have different blocking activities. Anti-PD-L1 mAb 10F.2H11 sterically and functionally blocks only PD-L1:B7-1 interactions, whereas anti-PD-L1 mAb 10F.9G2 blocks both PD-L1:B7-1 and PD-L1:programmed death-1 interactions. Both Abs had potent, yet distinct effects in accelerating diabetes in NOD mice: the single-blocker 10F.2H11 mAb was more effective at precipitating diabetes in older (13-wk-old) than in younger (6- to 7-wk-old) mice, whereas the dual-blocker 10F.9G2 mAb rapidly induced diabetes in NOD mice of both ages. Similarly, 10F.2H11 accelerated diabetes in recipients of T cells from diabetic, but not prediabetic mice, whereas 10F.9G2 was effective in both settings. Both anti-PD-L1 mAbs precipitated diabetes in adoptive transfer models of CD4(+) and CD8(+) T cell-driven diabetes. Taken together, these data demonstrate that the PD-L1:B7-1 pathway inhibits potentially pathogenic self-reactive effector CD4(+) and CD8(+) T cell responses in vivo, and suggest that the immunoinhibitory functions of this pathway may be particularly important during the later phases of diabetogenesis.


Predictors of ceasing or reducing statin medication following a large increase in the consumer copayment for medications: a retrospective observational study.
Previous Australian research has shown that following the 21% increase in patient copayments for medications on the Pharmaceutical Benefits Scheme (PBS) in 2005, the use of lipid-lowering therapy declined by 5%. This study aimed to determine the demographic and clinical characteristics of individuals who continued, reduced or ceased their use of statin medication in 2005. Retrospective observational study using routinely collected administrative data. We used pharmaceutical claims, hospital separations and mortality records from 2000 to 2005 for the Western Australian population. The cohort comprised stable users of statin medication in 2004. Based on changes in statin use between 2004 and 2005, we identified individuals who: 1) continued using statins; 2) reduced their use by ≥20%; or 3) ceased therapy for at least the first 6 months in 2005. Multivariate logistic regression models were used to determine whether the demographic and clinical characteristics of the three groups differed. There were 205 924 statin users identified in Western Australia as of December 2004. After the January 2005 Pharmaceutical Benefits Scheme (PBS) copayment increase, 3.2% of users ceased their regular statin therapy, 12.9% reduced statin use and 83.9% continued statin use. This represented a 2.1% increase in statin users reducing or ceasing therapy compared to 2004. Predictors of cessation and reduction of statin therapy included younger age, greater socio-economic disadvantage, residing in very remote areas, having general beneficiary status, being a new statin user, having no prior history of ischaemic heart disease, having no prior history of a coronary artery revascularisation procedure, taking no other cardiovascular medication or diabetic medication, taking an increased number of medications, and having a lower level of adherence to statin medication in 2004. Compared to 2004, an additional 2.1% of statin users reduced or discontinued medication use in 2005, which may be attributed to an increase in the medication copayment. Individuals with general beneficiary status, and younger and healthier people were at particular risk of cessation or reduction in statin use in 2005.


Global sensitivity analysis for systems with independent and/or correlated inputs.
The objective of a global sensitivity analysis is to rank the importance of the system inputs considering their uncertainty and the influence they have upon the uncertainty of the system output, typically over a large region of input space. This paper introduces a new unified framework of global sensitivity analysis for systems whose input probability distributions are independent and/or correlated. The new treatment is based on covariance decomposition of the unconditional variance of the output. The treatment can be applied to mathematical models, as well as to measured laboratory and field data. When the input probability distribution is correlated, three sensitivity indices give a full description, respectively, of the total, structural (reflecting the system structure) and correlative (reflecting the correlated input probability distribution) contributions for an input or a subset of inputs. The magnitudes of all three indices need to be considered in order to quantitatively determine the relative importance of the inputs acting either independently or collectively. For independent inputs, these indices reduce to a single index consistent with previous variance-based methods. The estimation of the sensitivity indices is based on a meta-modeling approach, specifically on the random sampling-high dimensional model representation (RS-HDMR). This approach is especially useful for the treatment of laboratory and field data where the input sampling is often uncontrolled.


Frequency and clinical impact of CDKN2A/ARF/CDKN2B gene deletions as assessed by in-depth genetic analyses in adult T cell acute lymphoblastic leukemia.
Recurrent deletions of the CDKN2A/ARF/CDKN2B genes encoded at chromosome 9p21 have been described in both pediatric and adult acute lymphoblastic leukemia (ALL), but their prognostic value remains controversial, with limited data on adult T-ALL. Here, we investigated the presence of homozygous and heterozygous deletions of the CDKN2A/ARF and CDKN2B genes in 64 adult T-ALL patients enrolled in two consecutive trials from the Spanish PETHEMA group. Alterations in CDKN2A/ARF/CDKN2B were detected in 35/64 patients (55%). Most of them consisted of 9p21 losses involving homozygous deletions of the CDKNA/ARF gene (26/64), as confirmed by single nucleotide polymorphism (SNP) arrays and interphase fluorescence in situ hybridization (iFISH). Deletions involving the CDKN2A/ARF/CDKN2B locus correlated with a higher frequency of cortical T cell phenotype and a better clearance of minimal residual disease (MRD) after induction therapy. Moreover, the combination of an altered copy-number-value (CNV) involving the CDKN2A/ARF/CDKN2B gene locus and undetectable MRD (≤ 0.01%) values allowed the identification of a subset of T-ALL with better overall survival in the absence of hematopoietic stem cell transplantation.


Panel discussion review: session two--interpretation of observed associations between multiple ambient air pollutants and health effects in epidemiologic analyses.
Air pollution epidemiologic research has often utilized ambient air concentrations measured from centrally located monitors as a surrogate measure of exposure to these pollutants. Associations between these ambient concentrations and health outcomes such as lung function, hospital admissions, and mortality have been examined in short- and long-term cohort studies as well as in time-series and case-crossover studies. The issues related to interpreting the observed associations of ambient air pollutants with health outcomes were discussed at the US EPA sponsored workshop on December 13 and 14, 2006 in Chapel Hill, North Carolina, USA. The second session of this workshop focused on the following topics: (1) statistical methodology and study designs that may improve understanding of multipollutant health effects; (2) ambient concentrations as surrogate measures of pollutant mixtures; and (3) source-focused epidemiologic research. New methodology and approaches to better distinguish the effects of individual pollutants include multicity hierarchical modeling and the use of case-crossover analysis to control for copollutants. An alternative approach is to examine the mixture as a whole using principal component analysis. Another important consideration is to what extent the observed health associations are attributable to individual pollutants, which are often from common sources and are correlated, versus the pollutant mixtures that the pollutants are representing. For example, several ambient air concentrations, such as particulate matter mass, nitrogen dioxide, and carbon monoxide, may be serving as surrogate measures of motor vehicle exhaust. Source apportionment analysis is one method that may allow further advancement in understanding the source components that contribute to multipollutant health effects.


Patterns of recurrence following liver resection for colorectal metastases: effect of primary rectal tumor site.
Patients with rectal adenocarcinoma are at increased risk of locoregional recurrence compared with patients with colon cancer. This may affect the pattern of recurrence and survival rates following hepatic resection of liver metastases from rectal adenocarcinoma. Retrospective review of a prospectively collected cancer center database. From April 1, 1984, to December 31, 2005, 582 patients with liver metastases from a primary colorectal adenocarcinoma underwent hepatic resection. Clinical and pathological factors were analyzed using Cox regression analyses and log-rank tests. Of 582 patients, 141 (24.2%) had liver metastases from a primary rectal tumor site. Treatment of the primary rectal tumor most frequently included chemoradiation therapy (59.6%) and low anterior resection (63.1%). Most rectal tumors were pathological stage T3/T4 (85.8%) and N1 (68.1%). Treatment directed at the hepatic metastases included resection only (81.5%), resection plus radiofrequency ablation (17.8%), or radiofrequency ablation only (0.7%). With a median follow-up time of 30.7 months, 80 of 141 patients (56.7%) developed recurrence; 23 patients (16.3%) developed recurrence in the pelvis. Of 23 patients with pelvic recurrence, 56.5% also developed recurrence in the liver. The 3- and 5-year survival rates for all patients were 62.4% and 36.4%, respectively. Of 80 patients who had a recurrence following hepatic metastectomy, 23 (28.8%) underwent another operation. Following repeat metastectomy, 3- and 5-year survival rates were 76.7% and 38.6%, respectively. Following resection of hepatic rectal metastases, pelvic recurrence is relatively common, and most patients with pelvic recurrence will also develop recurrence in the liver. Surgery for recurrent disease following hepatic resection of rectal metastases is warranted among well-selected patients.


Understanding the distinctive elastic constants in an oxadiazole bent-core nematic liquid crystal.
The splay and bend elastic constants of the bent-core oxadiazole material [C5-Ph-ODBP-Ph-OC12] have been investigated as a function of temperature across the nematic phase. The bend constant K(33) is found to take values of ~3.0 pN and to be almost temperature independent, whereas, the splay constant K(11) increases monotonically from ~3.5 pN close to the isotropic phase transition to values of ~9 pN deep in the nematic phase. No pretransitional divergence is observed in either K(11) or K(33) at temperatures approaching the underlying phase. This behavior of the elastic constants is distinct from that observed in rodlike liquid crystal systems but appears to share characteristics with the few other bent-core nematic systems studied to date. We discuss the interdependence of the elastic constants, the birefringence, and the order parameter to allow a comparison of the observed behavior with theory. We show that calculations of the elastic constants via molecular-field theory and atomistic modeling are in excellent qualitative as well as good quantitative (within 2 pN) agreement with the measurements across the temperature range, offering a deeper understanding of the elasticity in bent-core nematic materials than has been, hitherto, available.


Spatial mRNA Expression and Response to Fasting and Refeeding of Neutral Amino Acid Transporters slc6a18 and slc6a19a in the Intestinal Epithelium of Mozambique tilapia.
The mRNA expressions of the epithelial neutral amino acid transporters slc6a18 and slc6a19a in the five segments (HL, PMC, GL, DMC, and TS) of the intestine of Mozambique tilapia, and their responses to fasting and refeeding were investigated for a better understanding of the functional and nutritional characteristics of slc6a18 and slc6a19a. Although both slc6a18 and slc6a19a were expressed mainly in the intestine, these genes showed opposing spatial distributions along the intestine. The slc6a18 was mainly expressed in the middle (GL) and posterior (DMC and TS) intestines, while slc6a19a was specifically expressed in the anterior intestine (HL and PMC). Large decreases of amino acid concentrations from the HL to GL imply that amino acids are mainly absorbed before reaching the GL, suggesting an important role of slc6a19a in the absorption. Moreover, substantial amounts of some neutral amino acids with the isoelectric point close to 6 remain in the GL. These are most likely the remaining unabsorbed amino acids or those from of amino acid antiporters which release neutral amino acids in exchange for uptake of its substrates. These amino acids were diminished in the TS, suggesting active absorption in the posterior intestine. This suggests that slc6a18 is essential to complete the absorption of neutral amino acids. At fasting, significant downregulation of slc6a19a expression was observed from the initial up to day 2 and became stable from day 4 to day 14 in the HL and PMC suggesting that slc6a19a expression reflects nutritional condition in the intestinal lumen. Refeeding stimulates slc6a19a expression, although expressions did not exceed the initial level within 3 days after refeeding. The slc6a18 expression was decreased during fasting in the GL but no significant change was observed in the DMC. Only a transient decrease was observed at day 2 in the TS. Refeeding did not stimulate slc6a18 expression. Results in this study suggest that Slc6a18 and Slc6a19 have different roles in the intestine, and that both of these contribute to establish the efficient neutral amino acid absorption system in the tilapia.


Intraoperative patient selection for tubeless percutaneous nephrolithotomy.
This study was conducted to report our experience of intraoperative patient selection for tubeless percutaneous nephrolithotomy (PCNL) based on a tentative decision-making algorithm. Thirty-four consecutive patients who were scheduled to undergo tubeless PCNL were included and medical records were obtained from a prospectively maintained database for these patients. After completion of PCNL, the nephrostomy site was observed with a safety guidewire in place. If there was no significant bleeding through the tract, tubeless PCNL was performed, and in cases with significant bleeding or other complications, nephrostomy catheter insertion was performed as usual. In 29 cases (85.3%), tubeless PCNL was performed according to our decision-making protocol. Mean stone size was 7.33 ± 9.35 cm(2). Mean hospital stay was 2.61 ± 1.01 days. The difference between preoperative and postoperative hemoglobin was 0.68 ± 1.22 g/dL (p > 0.05). Visual analog pain scale scores immediately post-operation, on postoperative day one and on the day of discharge were 4.62 ± 1.80, 3.25 ± 1.68 (postoperative day one vs. operative day; p = 0.001), and 1.87 ± 0.83 (the day of discharge vs. operative day; p = 0.001), respectively. The success rate with insignificant remnant stones was 85.2% and complete stone-free rate was 76.5%. In conclusion, tubeless PCNL was performed successfully with low complication rate and reduced pain score through our decision-making algorithm.


Indirect capillary electrophoresis immunoassay of membrane protein in extracellular vesicles.
Extracellular vesicles (EVs) exist in biological fluids such as blood, urine, and cerebrospinal fluid, and these have shown promise for use as biomarkers of cancers. Conventional methods for determination of EVs include direct detection via enzyme-linked immunosorbent assay and detection of their membrane proteins via western blotting. These techniques, however, have individual shortcomings in terms of the need for large sample consumption, processes that are time-consuming, and a lack of the capacity for quantification. In this study, we developed a method to determine the EV membrane protein, CD63, by coupling capillary electrophoresis immunoassay with laser-induced fluorescence (CEIA-LIF). In this process, the EVs were isolated from a culture medium and were subsequently reacted with a fluorescently labeled anti-CD63 antibody to form a CD63 complex localized on the surface of EVs. After removing the EVs containing the CD63 immune complex by centrifugation, the supernatant containing the free fluorescent antibody was injected into a capillary to serve as a sample. A decrease in the peak area of the free fluorescent antibody became apparent when the amount of EVs was increased while that of the fluorescent antibody remained constant. The peak areas were decreased proportionally against the increased amounts of EVs. The concentration of the CD63 could then be estimated based on the slope of the linear relationship. This study is the first to quantify CD63 immobilized on EVs via CEIA-LIF, which is a novel method with the potential to determine membrane proteins localized on the surface of EVs.


Depression, marital satisfaction and communication in couples: investigating gender differences.
The correlation between depression and dysfunctional marital interaction is well documented, but only a few studies have examined gender-related differences in marital interaction patterns of couples with a depressed partner. In this paper we examined differences in observed marital communication in a sample of 62 Swiss couples presenting for treatment of depression. There were 16 maritally distressed couples with a depressed wife, 21 maritally nondistressed couples with a depressed wife, 18 maritally distressed couples with a depressed husband, and 7 maritally nondistressed couples with a depressed husband. Marital interaction behavior was found to depend on gender, depression, marital distress, as well as gender of the depressed partner. Our results suggest the need for a gender-sensitive model of the link between marital interaction and depression.


Oriented aggregation of lepidocrocite and impact on surface charge development.
The impact of lepidocrocite (γ-FeOOH) nanoparticle aggregation on mineral surface charge development was resolved in aqueous solutions of NaCl and NaClO4. Synthetic rod-like particles exhibiting charged edge (100) and neutrally/low-charged (010) faces self-aggregated in salt-free solutions. Aggregation was notably imaged by high-resolution transmission electron microscopy, and inferred by decreases in N2(g)-B.E.T. specific surface area from 94 m(2)/g to 77 m(2)/g after 12 months, and to 66 m(2)/g after 33 months storage. Potential determining (H(+), OH(-)) ions loadings in the 4-11 pH range were unchanged only if the particles remained aggregated in NaCl but only if they were disaggregated in NaClO4. These differences, alongside molecular simulations and experimental ion loadings resolved in other studies from our group, point to important controls on background electrolyte ion identity on the aggregation and charge development in lepidocrocite. These results may apply further to other mineral surfaces of comparable surface (hydr)oxo populations.


High-Throughput Screening Approach for the Optoelectronic Properties of Conjugated Polymers.
We propose a general high-throughput virtual screening approach for the optical and electronic properties of conjugated polymers. This approach makes use of the recently developed xTB family of low-computational-cost density functional tight-binding methods from Grimme and co-workers, calibrated here to (Time-Dependent) Density Functional Theory ((TD)DFT) data computed for a representative diverse set of (co)polymers. Parameters drawn from the resulting calibration using a linear model can then be applied to the xTB derived results for new polymers, thus generating near DFT-quality data with orders of magnitude reduction in computational cost. As a result, after an initial computational investment for calibration, this approach can be used to quickly and accurately screen on the order of thousands of polymers for target applications. We also demonstrate that the (opto)electronic properties of the conjugated polymers show only a very minor variation when considering different conformers and that the results of high-throughput screening are therefore expected to be relatively insensitive with respect to the conformer search methodology applied.


The combined treatment of praziquantel with osteopontin immunoneutralization reduces liver damage in Schistosoma japonicum-infected mice.
The aim of this study was to evaluate the therapeutic effects of osteopontin neutralization treatment on schistosome-induced liver injury in BALB/C mice. We randomly divided 100 BALB/C mice into groups A, B, C, D and group E. Mice in all groups except group A were abdominally infected with schistosomal cercariae to induce a schistosomal hepatopathological model. Mice in group C, D and group E were respectively administered with praziquantel, praziquantel plus colchicine and praziquantel plus neutralizing osteopontin antibody. We extracted mouse liver tissues at 3 and 9 weeks after the 'stool-eggs-positive' day, observed liver histopathological changes by haematoxylin-eosin and Masson trichrome staining and detected the expression of osteopontin, alpha-smooth muscle actin (α-SMA) and transforming growth factor-beta (TGF-β1) by immunohistochemistry, RT-PCR and Western blot. We found that praziquantel plus neutralizing osteopontin antibody treatment significantly decreased the granuloma dimension, the percentage of collagen and the expression of osteopontin, α-SMA and TGF-β1 compared to praziquantel plus colchicine treatment in both the acute and chronic stage of schistosomal liver damage (P<0·05). So we believe that the combined regimen of osteopontin immunoneutralization and anti-helminthic treatment can reduce the granulomatous response and liver fibrosis during the schistosomal hepatopathologic course.


Effect of prolonged wakefulness on electroencephalographic oscillatory activity during sleep.
The human sleep electroencephalogram (EEG) is characterized by the occurrence of distinct oscillatory events such as delta waves, sleep spindles and alpha activity. We applied a previously proposed algorithm for the detection of such events and investigated their incidence and frequency in baseline and recovery sleep after 40 h of sustained wakefulness in 27 healthy young subjects. The changes in oscillatory events induced by sleep deprivation were compared to the corresponding spectral changes. Both approaches revealed, on average, an increase in low frequency activity and a decrease in spindle activity after sleep deprivation. However, the increase of oscillatory events in the delta range and decrease in the sigma range occurred in a more restricted frequency range compared to spectral changes. The mean relative power spectra showed a significant increase in theta and alpha activity after sleep deprivation while, on average, the event analysis showed only a weak effect in the theta band. The reason for this discrepancy is that the spectral analysis does not distinguish between diffuse activity and clearly visible temporally localized oscillations, while the event analysis would detect only the latter. Additionally, only a few individuals clearly showed activity in the theta or alpha frequency bands. Conversely, event analysis revealed that some individuals showed an increased rate of sleep spindles after sleep deprivation, a fact that was not evident in the relative power spectra due to a decrease in background activity. The two methods complement each other and facilitate the interpretation of distinct changes induced by prolonged wakefulness in sleep EEG.


Flexible work in call centres: Working hours, work-life conflict & health.
Call-centre workers encounter major psychosocial pressures, including high work intensity and undesirable working hours. Little is known, however, about whether these pressures vary with employment status and how they affect work-life conflict and health. Questionnaire data were collected from 179 telephone operators in Sydney, Australia, of whom 124 (69.3%) were female and 54 (30.2%) were male. Ninety-three (52%) were permanent full-time workers, 37 (20.7%) were permanent part-time, and 49 (27.4%) were casual employees. Hypothesised structural relationships between employment status, working hours and work organisation, work-life conflict and health were tested using partial least squares modelling in PLS (Chin, 1998). The final model demonstrated satisfactory fit. It supported important elements of the hypothesised structure, although four of the proposed paths failed to reach significance and the fit was enhanced by adding a path. The final model indicated that casual workers reported more variable working hours which were relatively weakly associated with greater dissatisfaction with hours. The interaction of schedule control and variability of hours also predicted dissatisfaction with hours. Conversely, permanent workers reported greater work intensity, which was associated with both lower work schedule control and greater work-life conflict. Greater work-life conflict was associated with more fatigue and psychological symptoms. Labour market factors and the undesirability of longer hours in a stressful, high-intensity work environment appear to have contributed to the results.


Analysis of a human V beta gene subfamily.
We have isolated and sequenced five germline V beta gene segments that are homologous to the V region of the YT35 cDNA encoding the beta chain of the T cell antigen receptor from the tumor MOLT-3. One of these gene segments is identical to the YT35 V segment, and therefore is the corresponding germline V beta gene segment encoding the YT35 cDNA. The other four V beta members exhibit 77-98% homology to the YT35 V gene segment. Two of these V beta gene segments are pseudogenes. Analyses of the coding region sequences reveal that, although the V beta segments are very diverse, they are mutating at a rate comparable to that observed in most eukaryotic genes. Analyses of the genomic clones show that the spacing distance between germline V beta gene segments ranges from 3 kb to greater than 30 kb, and the entire V beta 8 subfamily appears to be linked by a total of no more than 110 kb of DNA.


Adipocyte-macrophage interaction may mediate LPS-induced low-grade inflammation: potential link with metabolic complications.
Chronic low-grade infection has been suggested to be associated with metabolic disorder such as diabetes. However, the molecular mechanism underlying this important association is largely unknown. The only clue established so far is that many subjects exhibit elevated levels of C-reactive protein as measured by highly sensitive assay. Here, we hypothesized that adipocyte-macrophage interaction plays a key role in amplifying such low grade infection to the level of influencing metabolic disorders. The presence of macrophages in abdominal adipose tissues was investigated by immunohistochemistry. To see whether molecules associated with acute phase protein, LPS signaling, and persistent recruitment of monocytes, are produced at higher amounts in adipocytes co-cultured with macrophages stimulated with low concentration of LPS (1 ng/ml), we measured serum amyloid A (SAA), LPS binding protein (LBP), soluble CD14 (sCD14), and RANTES levels in culture supernatant of co-cultures. Lastly, we investigated in vivo effect of low-grade LPS infusion on the production of these molecules using obese model mice. The macrophages were certainly identified in abdominal adipose tissues. Investigated molecules, especially LBP, SAA, and RANTES were produced at higher amounts in co-cultures stimulated with LPS compared with the cells without LPS. The ob/ob, and high-fat diet-induced obesity mice produced higher amounts of LBP, SAA, and RANTES one day after LPS infusion (1 ng/ml/g body weight) compared with ob/- and normal-fat fed control mice. Thus, adipocytes and infiltrated macrophages, and their interaction with low endotoxin stimulation appear to play an important role in amplifying and maintaining LPS-induced low-grade inflammation.


Immunodeficiency virus uptake, turnover, and 2-phase transfer in human dendritic cells.
HIV-1 subverts antigen processing in dendritic cells (DCs) resulting in viral uptake, infection, and transfer to T cells. Although DCs bound monomeric gp120 and HIV-1 similarly, virus rarely colocalized with endolysosomal markers, unlike gp120, suggesting HIV-1 alters endolysosomal trafficking. Virus within DC intracellular compartments rapidly moved to DC-CD4+ lymphocyte synapses when introduced to CD4+ lymphocyte cultures. Although viral harboring and transfer from nonlysosomal compartments was transient, given DC-associated virus protein, nucleic acids, and infectious HIV-1 transfer to CD4+, lymphocytes decayed within 24 hours. However a second long-term transfer phase was apparent in immature DCs after 48 hours as a zidovudine-sensitive rise in proviral DNA. Therefore, DCs transfer HIV-1 to CD4+ lymphocytes in 2 distinct phases. Immature and mature DCs first divert virus from the endolysosomal pathway to the DC-T-cell synapse. Secondly, the later transfer phase from immature DCs is through de novo HIV-1 production. Thus, the controversy of DCs being infected or not infected for the mechanics of viral transfer to CD4+ lymphocytes can be addressed as a function of time.


Gender Differences in Weight-Related Attitudes and Behaviors Among Overweight and Obese Adults in the United States.
Few studies have used nationally representative data to focus specifically on gender differences in weight-related outcomes. This article examines gender differences in weight-related outcomes across the body mass index (BMI) spectrum in overweight and obese adults. Data from the National Health and Nutrition Examination Survey 2009-2010 was analyzed. Weight-related outcomes were accurate weight perception, weight dissatisfaction, attempted weight loss, successful weight loss, and weight loss strategies. Compared with women, overweight and obese men were less likely to have accurate weight perception (odds ratio [OR] = 0.36; 95% confidence interval [CI] = 0.30-0.44), weight dissatisfaction (OR = 0.39; 95% CI = 0.32-0.47), and attempted weight loss (OR = 0.55; 95% CI = 0.48-0.63). The modifying effect of gender on these associations decreased as BMI increased. By BMI 35, the mean probability of women and men to have accurate weight perception and weight dissatisfaction was 90%; attempted weight loss was 60% (women) and 50% (men). At lower BMIs, men had up to 40% less probability than women for these weight loss outcomes. Men who attempted weight loss were more likely than women to lose and maintain ≥10 lb over 1 year (OR = 1.41; 95% CI = 1.20-1.65) and increase exercise and eat less fat as weight loss strategies; women were more likely to join weight loss programs, take prescription diet pills, and follow special diets. A need exists for male-specific interventions to improve overweight and obese men's likelihood for accurate weight perception, attempted weight loss, and ultimately, successful weight loss.


Toxoplasma gondii infection in the mountain hare (Lepus timidus) and domestic rabbit (Oryctolagus cuniculus). II. Early immune reactions.
As already reported, the mountain hare is much more susceptible than the domestic rabbit to oral inoculation with Toxoplasma gondii, as judged by pathological changes and dissemination of parasites within the body. In the present paper, further interspecies variations are reported. Concentrations of the acute phase reactant haptoglobin were raised in hares but not in rabbits one week post-infection (pi), probably reflecting the severe tissue damage present. No difference in the early humoral immune response of hares and rabbits was found, both species producing IgM and IgG antibodies to T. gondii one week pi. Lymphocyte stimulation tests performed before and one week after inoculation showed a high proliferative response to the parasite in blood cell cultures from rabbits but not hares. The fatal outcome of T. gondii infection in the hares is probably due, at least in part, to the lack of cellular response.


Nile red: a selective fluorescent stain for intracellular lipid droplets.
We report that the dye nile red, 9-diethylamino-5H-benzo[alpha]phenoxazine-5-one, is an excellent vital stain for the detection of intracellular lipid droplets by fluorescence microscopy and flow cytofluorometry. The specificity of the dye for lipid droplets was assessed on cultured aortic smooth muscle cells and on cultured peritoneal macrophages that were incubated with acetylated low density lipoprotein to induce cytoplasmic lipid overloading. Better selectivity for cytoplasmic lipid droplets was obtained when the cells were viewed for yellow-gold fluorescence (excitation, 450-500 nm; emission, greater than 528 nm) rather than red fluorescence (excitation, 515-560 nm; emission, greater than 590 nm). Nile red-stained, lipid droplet-filled macrophages exhibited greater fluorescence intensity than did nile red-stained control macrophages, and the two cell populations could be differentiated and analyzed by flow cytofluorometry. Such analyses could be performed with either yellow-gold or red fluorescence, but when few lipid droplets per cell were present, the yellow-gold fluorescence was more discriminating. Nile red exhibits properties of a near-ideal lysochrome. It is strongly fluorescent, but only in the presence of a hydrophobic environment. The dye is very soluble in the lipids it is intended to show, and it does not interact with any tissue constituent except by solution. Nile red can be applied to cells in an aqueous medium, and it does not dissolve the lipids it is supposed to reveal.


Disruption of the presynaptic cytomatrix protein bassoon degrades ribbon anchorage, multiquantal release, and sound encoding at the hair cell afferent synapse.
Inner hair cells (IHCs) of the cochlea use ribbon synapses to transmit auditory information faithfully to spiral ganglion neurons (SGNs). In the present study, we used genetic disruption of the presynaptic scaffold protein bassoon in mice to manipulate the morphology and function of the IHC synapse. Although partial-deletion mutants lacking functional bassoon (Bsn(ΔEx4/5)) had a near-complete loss of ribbons from the synapses (up to 88% ribbonless synapses), gene-trap mutants (Bsn(gt)) showed weak residual expression of bassoon and 56% ribbonless synapses, whereas the remaining 44% had a loosely anchored ribbon. Patch-clamp recordings and synaptic CaV1.3 immunolabeling indicated a larger number of Ca(2+) channels for Bsn(gt) IHCs compared with Bsn(ΔEx4/5) IHCs and for Bsn(gt) ribbon-occupied versus Bsn(gt) ribbonless synapses. An intermediate phenotype of Bsn(gt) IHCs was also found by membrane capacitance measurements for sustained exocytosis, but not for the size of the readily releasable vesicle pool. The frequency and amplitude of EPSCs were reduced in Bsn(ΔEx4/5) mouse SGNs, whereas their postsynaptic AMPA receptor clusters were largely unaltered. Sound coding in SGN, assessed by recordings of single auditory nerve fibers and their population responses in vivo, was similarly affected in Bsn(gt) and Bsn(ΔEx4/5) mice. Both genotypes showed impaired sound onset coding and reduced evoked and spontaneous spike rates. In summary, reduced bassoon expression or complete lack of full-length bassoon impaired sound encoding to a similar extent, which is consistent with the comparable reduction of the readily releasable vesicle pool. This suggests that the remaining loosely anchored ribbons in Bsn(gt) IHCs were functionally inadequate or that ribbon independent mechanisms dominated the coding deficit.


Chromosome Painting Facilitates Anchoring Reference Genome Sequence to Chromosomes In Situ and Integrated Karyotyping in Banana (Musa Spp.).
Oligo painting FISH was established to identify all chromosomes in banana (Musa spp.) and to anchor pseudomolecules of reference genome sequence of Musa acuminata spp. malaccensis "DH Pahang" to individual chromosomes in situ. A total of 19 chromosome/chromosome-arm specific oligo painting probes were developed and were shown to be suitable for molecular cytogenetic studies in genus Musa. For the first time, molecular karyotypes of diploid M. acuminata spp. malaccensis (A genome), M. balbisiana (B genome), and M. schizocarpa (S genome) from the Eumusa section of Musa, which contributed to the evolution of edible banana cultivars, were established. This was achieved after a combined use of oligo painting probes and a set of previously developed banana cytogenetic markers. The density of oligo painting probes was sufficient to study chromosomal rearrangements on mitotic as well as on meiotic pachytene chromosomes. This advance will enable comparative FISH mapping and identification of chromosomal translocations which accompanied genome evolution and speciation in the family Musaceae.


S100A7-downregulation inhibits epidermal growth factor-induced signaling in breast cancer cells and blocks osteoclast formation.
S100A7 is a small calcium binding protein, which has been shown to be differentially expressed in psoriatic skin lesions, as well as in squamous cell tumors of the skin, lung and breast. Although its expression has been correlated to HER+ high-grade tumors and to a high risk of progression, the molecular mechanisms of these S100A7-mediated tumorigenic effects are not well known. Here, we showed for the first time that epidermal growth factor (EGF) induces S100A7 expression in both MCF-7 and MDA-MB-468 cell lines. We also observed a decrease in EGF-directed migration in shRNA-downregulated MDA-MB-468 cell lines. Furthermore, our signaling studies revealed that EGF induced simultaneous EGF receptor phosphorylation at Tyr1173 and HER2 phosphorylation at Tyr1248 in S100A7-downregulated cell lines as compared to the vector-transfected controls. In addition, reduced phosphorylation of Src at tyrosine 416 and p-SHP2 at tyrosine 542 was observed in these downregulated cell lines. Further studies revealed that S100A7-downregulated cells had reduced angiogenesis in vivo based on matrigel plug assays. Our results also showed decreased tumor-induced osteoclastic resorption in an intra-tibial bone injection model involving SCID mice. S100A7-downregulated cells had decreased osteoclast number and size as compared to the vector controls, and this decrease was associated with variations in IL-8 expression in in vitro cell cultures. This is a novel report on the role of S100A7 in EGF-induced signaling in breast cancer cells and in osteoclast formation.


Aboriginal and Non-Aboriginal Emergency Department Presentations Involving Suicide-Related Thoughts and Behaviors.
Background: Rates of hospital admission for suicide-related thoughts and behaviors (SRTBs) are elevated in the Northern Territory (NT) of Australia, especially by Aboriginal people, but very little is known about emergency department (ED) presentations. Aim: We aimed to profile ED presentations in the NT involving SRTBs by Indigenous status and compare discharge arrangements. Method: Logistic regression analyses were performed on data from electronic patient records of consecutive ED presentations involving SRTBs. Results: During the study period, 167 presentations were observed. Aboriginal patients were more likely to present from remote areas and to report substance misuse and family conflict or violence compared with non-Aboriginal patients. In both groups, males were more likely than females to be admitted as were persons presenting with self-harm compared with those who had suicidal thoughts only. No differences in discharge arrangements were identified by Indigenous status. Limitations: The small scale of the study and use of administrative records points to the need for further research to improve the quality of the evidence. Conclusion: While presentations by high-risk groups are more likely to be admitted for further care, the assessment of psychosocial risks and needs in EDs is vital to informing decisions for aftercare that support recovery in the community for Aboriginal patients and patients discharged from EDs.


Smoking trajectories among Koreans in Seoul and California: exemplifying a common error in age parameterization.
Immigration to a nation with a stronger anti-smoking environment has been hypothesized to make smoking less common. However, little is known about how environments influence risk of smoking across the lifecourse. Research suggested a linear decline in smoking over the lifecourse but these associations, in fact, might not be linear. This study assessed the possible nonlinear associations between age and smoking and examined how these associations differed by environment through comparing Koreans in Seoul, South Korea and Korean Americans in California, United States. Data were drawn from population based telephone surveys of Korean adults in Seoul (N=500) and California (N=2,830) from 2001-2002. Locally weighted scatterplot smoothing (lowess) was used to approximate the association between age and smoking with multivariable spline logistic regressions, including adjustment for confounds used to draw population inferences. Smoking differed across the lifecourse between Korean and Korean American men. The association between age and smoking peaked around 35 years among Korean and Korean American men. From 18 to 35 the probability of smoking was 57% higher (95%CI, 40 to 71) among Korean men versus 8% (95%CI, 3 to 19) higher among Korean American men. A similar difference in age after 35, from 40 to 57 years of age, was associated with a 2% (95%CI, 0 to 10) and 20% (95%CI, 16 to 25) lower probability of smoking among Korean and Korean American men. A nonlinear pattern was also observed among Korean American women. Social role transitions provide plausible explanations for the decline in smoking after 35. Investigators should be mindful of nonlinearities in age when attempting to understand tobacco use.


Cross talk between 2,4-diacetylphloroglucinol-producing biocontrol pseudomonads on wheat roots.
The performance of Pseudomonas biocontrol agents may be improved by applying mixtures of strains which are complementary in their capacity to suppress plant diseases. Here, we have chosen the combination of Pseudomonas fluorescens CHA0 with another well-characterized biocontrol agent, P. fluorescens Q2-87, as a model to study how these strains affect each other's expression of a biocontrol trait. In both strains, production of the antimicrobial compound 2,4-diacetylphloroglucinol (DAPG) is a crucial factor contributing to the suppression of root diseases. DAPG acts as a signaling compound inducing the expression of its own biosynthetic genes. Experimental setups were developed to investigate whether, when combining strains CHA0 and Q2-87, DAPG excreted by one strain may influence expression of DAPG-biosynthetic genes in the other strain in vitro and on the roots of wheat. DAPG production was monitored by observing the expression of lacZ fused to the biosynthetic gene phlA of the respective strain. Dual-culture assays in which the two strains were grown in liquid medium physically separated by a membrane revealed that Q2-87 but not its DAPG-negative mutant Q2-87::Tn5-1 strongly induced phlA expression in a DeltaphlA mutant of strain CHA0. In the same way, phlA expression in a Q2-87 background was induced by DAPG produced by CHA0. When coinoculated onto the roots of wheat seedlings grown under gnotobiotic conditions, strains Q2-87 and CHA0, but not their respective DAPG-negative mutants, were able to enhance phlA expression in each other. In summary, we have established that two nonrelated pseudomonads may stimulate each other in the expression of an antimicrobial compound important for biocontrol. This interpopulation communication occurs in the rhizosphere, i.e., at the site of pathogen inhibition, and is mediated by the antimicrobial compound itself acting as a signal exchanged between the two pseudomonads.


Reduced lymphocyte longevity and homeostatic proliferation in lamin B receptor-deficient mice results in profound and progressive lymphopenia.
The lamin B receptor (LBR) is a highly unusual inner nuclear membrane protein with multiple functions. Reduced levels are associated with decreased neutrophil lobularity, whereas complete absence of LBR results in severe skeletal dysplasia and in utero/perinatal lethality. We describe a mouse pedigree, Lym3, with normal bone marrow and thymic development but profound and progressive lymphopenia particularly within the T cell compartment. This defect arises from a point mutation within the Lbr gene with only trace mutant protein detectable in homozygotes, albeit sufficient for normal development. Reduced T cell homeostatic proliferative potential and life span in vivo were found to contribute to lymphopenia. To investigate the role of LBR in gene silencing in hematopoietic cells, we examined gene expression in wild-type and mutant lymph node CD8 T cells and bone marrow neutrophils. Although LBR deficiency had a very mild impact on gene expression overall, for common genes differentially expressed in both LBR-deficient CD8 T cells and neutrophils, gene upregulation prevailed, supporting a role for LBR in their suppression. In summary, this study demonstrates that LBR deficiency affects not only nuclear architecture but also proliferation, cell viability, and gene expression of hematopoietic cells.


Antibodies to influenza nucleoprotein cross-react with human hypocretin receptor 2.
The sleep disorder narcolepsy is linked to the HLA-DQB1*0602 haplotype and dysregulation of the hypocretin ligand-hypocretin receptor pathway. Narcolepsy was associated with Pandemrix vaccination (an adjuvanted, influenza pandemic vaccine) and also with infection by influenza virus during the 2009 A(H1N1) influenza pandemic. In contrast, very few cases were reported after Focetria vaccination (a differently manufactured adjuvanted influenza pandemic vaccine). We hypothesized that differences between these vaccines (which are derived from inactivated influenza viral proteins) explain the association of narcolepsy with Pandemrix-vaccinated subjects. A mimic peptide was identified from a surface-exposed region of influenza nucleoprotein A that shared protein residues in common with a fragment of the first extracellular domain of hypocretin receptor 2. A significant proportion of sera from HLA-DQB1*0602 haplotype-positive narcoleptic Finnish patients with a history of Pandemrix vaccination (vaccine-associated narcolepsy) contained antibodies to hypocretin receptor 2 compared to sera from nonnarcoleptic individuals with either 2009 A(H1N1) pandemic influenza infection or history of Focetria vaccination. Antibodies from vaccine-associated narcolepsy sera cross-reacted with both influenza nucleoprotein and hypocretin receptor 2, which was demonstrated by competitive binding using 21-mer peptide (containing the identified nucleoprotein mimic) and 55-mer recombinant peptide (first extracellular domain of hypocretin receptor 2) on cell lines expressing human hypocretin receptor 2. Mass spectrometry indicated that relative to Pandemrix, Focetria contained 72.7% less influenza nucleoprotein. In accord, no durable antibody responses to nucleoprotein were detected in sera from Focetria-vaccinated nonnarcoleptic subjects. Thus, differences in vaccine nucleoprotein content and respective immune response may explain the narcolepsy association with Pandemrix.


Dual Role of gnaA in Antibiotic Resistance and Virulence in Acinetobacter baumannii.
Acinetobacter baumannii is an important Gram-negative pathogen in hospital-related infections. However, treatment options for A. baumannii infections have become limited due to multidrug resistance. Bacterial virulence is often associated with capsule genes found in the K locus, many of which are essential for biosynthesis of the bacterial envelope. However, the roles of other genes in the K locus remain largely unknown. From an in vitro evolution experiment, we obtained an isolate of the virulent and multidrug-resistant A. baumannii strain MDR-ZJ06, called MDR-ZJ06M, which has an insertion by the ISAba16 transposon in gnaA (encoding UDP-N-acetylglucosamine C-6 dehydrogenase), a gene found in the K locus. The isolate showed an increased resistance toward tigecycline, whereas the MIC decreased in the case of carbapenems, cephalosporins, colistin, and minocycline. By using knockout and complementation experiments, we demonstrated that gnaA is important for the synthesis of lipooligosaccharide and capsular polysaccharide and that disruption of the gene affects the morphology, drug susceptibility, and virulence of the pathogen.


Anterior cage dislodgement in posterior lumbar interbody fusion: a review of 12 patients.
OBJECTIVE Interbody fusion cages are widely used to achieve initial fixation and secure spinal fusion; however, there are certain technique-related complications. Although anterior cage dislodgement can cause major vascular injury, the incidence is extremely rare. Here, the authors performed a review of anterior cage dislodgement following posterior lumbar interbody fusion (PLIF) surgery. METHODS The authors retrospectively reviewed the cases of 4625 patients who had undergone PLIF at 6 institutions between December 2007 and March 2015. They investigated the incidence and causes of surgery-related anterior cage dislodgement, salvage mechanisms, and postoperative courses. RESULTS Anterior cage dislodgement occurred in 12 cases (0.26%), all of which were caused by technical errors. In 9 cases, excessive cage impaction resulted in dislodgement. In 2 cases, when the cage on the ipsilateral side was inserted, it interacted and pushed out the other cage on the opposite side. In 1 case, the cage was positioned in an extreme lateral and anterior part of the intervertebral disc space, and it postoperatively dislodged. In 3 cases, the cage was removed in the same operative field. In the remaining 9 cases, CT angiography was performed postoperatively to assess the relationship between the dislodged cage and large vessels. Dislodged cages were conservatively observed in 2 cases. In 7 cases, the cage was removed because it was touching or compressing large vessels, and an additional anterior approach was selected. In 2 patients, there was significant bleeding from an injured inferior vena cava. There were no further complications or sequelae associated with the dislodged cages during the follow-up period. CONCLUSIONS Although rare, iatrogenic anterior cage dislodgement following a PLIF can occur. The authors found that technical errors made by experienced spine surgeons were the main causes of this complication. To prevent dislodgement, the surgeon should be cautious when inserting the cage, avoiding excessive cage impaction and ensuring cage control. Once dislodgement occurs, the surgeons must immediately address this difficult complication. First, the possibility of a large vessel injury should be considered. If the patient's vital signs are stable, the surgeon should continue with the surgery without cage removal and perform CT angiography postoperatively to assess the cage location. Blind maneuvers should be avoided when the surgical site cannot be clearly viewed. When the cage compresses or touches the aortic artery or vena cava, it is better to remove the cage to avoid late-onset injury to major vessels. When the cage does not compress or touch vessels, its removal is controversial. The risk factors associated with performing another surgery should be evaluated on a case-by-case basis.


Disulfide bridge conformers of mature BMP are inhibitors for heterotopic ossification.
Heterotopic ossification is a frequent complication in patients who have suffered head and neck traumas or undergone total hip replacement. Heterotopic ossification occurs when osteogenic precursor cells present at the ectopic site receive the necessary signal(s) to differentiate into osteoblasts. At the protein level, the key factors in differentiation of cells to the osteogenic lineage are BMPs. Stable BMP variants derived from the identical amino acid sequence but with different disulfide bridge configurations have been investigated and found to be capable of inhibiting ossification in vitro and in vivo in rodents. These findings provide a concept for the straightforward development of a novel class of BMP antagonists that could lead to new treatments for traumatically and genetically induced heterotopic ossification and also, possibly, for disorders in which other members of the TGF-beta superfamily are involved.


Binding preference of p62 towards LC3-ll during dopaminergic neurotoxin-induced impairment of autophagic flux.
Accumulating evidence has revealed that autophagy may be beneficial for treatment of neurodegenerative diseases through removal of abnormal protein aggregates. However, the critical autophagic events during neurodegeneration remain to be elucidated. Here, we investigated whether prototypic autophagic events occur in the MN9D dopaminergic neuronal cell line upon exposure to N-methyl-4-phenylpyridinium (MPP (+) ), a well-known dopaminergic neurotoxin. MPP (+) treatment induced both morphological and biochemical characteristics of autophagy, such as accumulation of autophagic vacuoles and LC3-II form and decreased p62 levels. Further investigation revealed that these phenomena were largely the consequences of blocked autophagic flux. Following MPP (+) treatment, levels of LC3-II formed and p62 dramatically increased in the Triton X-100-insoluble fraction. Levels of ubiquitinated proteins also increased in this fraction. Further colocalization analyses revealed that the punctated spots positive for both p62 and LC3 were more intense following MPP (+) treatment, suggesting drug-induced enrichment of these two proteins in the insoluble fraction. Intriguingly, reciprocal immunoprecipitation analysis revealed that p62 mainly precipitated with LC3-II form following MPP (+) treatment. Transient transfection of the mutant form of Atg4B, Atg4B (C74A) , which inhibits LC3 processing, dramatically decreased binding between p62 and LC3-II form. Taken together, our results indicate that p62 can be efficiently localized to autophagic compartments via preferential binding with LC3-II form. This colocalization may assist in removal of detergent-insoluble forms of damaged cellular proteins during dopaminergic neurotoxin-induced impairment of autophagic flux.


Novel SNPs in cytochrome P450 oxidoreductase.
Cytochrome P450 oxidoreductase (POR) is the single flavoprotein which donates electrons to the microsomal cytochrome P450 enzymes for oxidation of their substrates. In this study, we sequenced all 15 exons and the surrounding intronic sequences of POR in 100 human liver samples to identify novel and confirm known genetic polymorphisms in POR. Thirty-four single nucleotide polymorphisms (SNPs) were identified including 9 in the coding exons (5 synonymous and 4 nonsynonymous), 20 in the intronic regions, and 5 in the 3'-UTR. Of these, 9 were novel SNPs, including three nonsynonymous SNPs, SNH313003 (817733G>C; K49N), SNH313020 (848661C>A; L420M), and SNH313029 (849577T>C; L577P) with minor allele frequencies of 0.005, 0.045, and 0.020, respectively. We also confirmed a previously reported non-synonymous SNP rs1057868 (A503V) as well as five synonymous SNPs (G5G, T29T, P129P, S485S, and S572S) all with allele frequencies similar to those previously reported. Structurally, these polymorphisms occur in different regions: SNH313003 (K49N) in the amino-terminal tail, SNH313020 (L420M) in the connecting domain, SNH313029 (L577P) in the NADPH-binding domain, and rs1057868 (A503V) in the FAD binding domain.


[Relations between official and private veterinary services in epidemiology and the control of contagious diseases].
Growing budget restrictions in many countries have meant that official Veterinary Services cannot assume responsibility for any new activities. The natural reaction is to turn to private veterinary services to provide the support needed to strengthen the control and surveillance of priority diseases and thereby support the development of the livestock sector and the establishment of safe international trade. In this context, official Veterinary Services must work together with private veterinarians, delegating various technical animal health activities, so that they may focus their efforts on those tasks that cannot be delegated: standardisation, control, auditing, general system co-ordination, epidemiological surveillance, etc., as well as organising veterinary policy in order to make best use of budget resources. For these relations to be efficient, a dynamic, two-way epidemiological information mechanism must be created, whereby private veterinarians periodically keep governments informed, on the basis of an agreed methodology. Moreover, the official Veterinary Services must systematically transmit information on List A and B diseases of the OIE (World organisation for animal health), and perform detailed analyses of epidemiologically significant events. The article proposes the establishment of relations between public and private veterinary services as a way in which to provide the livestock sector with the health and hygiene conditions that are necessary for effective disease control, which in turn provides greater security for international trade and increased consumer protection.