Bacteraemia during tonsillectomy: a study of the factors involved and clinical implications.
Post-tonsillectomy bacteremia is a well-recognized aetiological factor in streptococcal endocarditis, and prophylactic penicillin has been recommended to reduce its incidence in susceptible patients undergoing tonsillectomy. Recent studies have shown a change in the microflora and an increase in the number of penicillin-resistant organisms in the tonsils of patients undergoing tonsillectomy. The aim of this study was to assess the incidence of post-tonsillectomy bacteraemia, to identify the micro-organisms associated with it and to review the suitability of penicillin in prophylactic regimens. The relationship between positive blood cultures and several clinical parameters such as fever, vomiting, pharyngeal discomfort, or dysphagia was also analysed. Of the 102 patients included in the study, 41 (40.1%) had positive post-tonsillectomy blood cultures. Haemophilus influenzae were isolated from 23 (56%) of the positive cultures and Streptococcus viridans in 15 (36.5%). Twenty-five per cent of H. influenzae produced beta-lactamase and only 30% of streptococci of the viridans group were penicillin-sensitive. Positivity of the blood cultures was not related to fever, discomfort, surgical technique, type of tonsil, or any of the parameters studied. Bacteraemia seems to be related to traction of the tonsil before dissection rather than direct spread of bacteria into the opened vessels. Using a beta-lactamase stable antibiotic instead of penicillin for prophylaxis would be more appropriate.


States of phage T3/T7 capsids: buoyant density centrifugation and cryo-EM.
Mature double-stranded DNA bacteriophages have capsids with symmetrical shells that typically resist disruption, as they must to survive in the wild. However, flexibility and associated dynamism assist function. We describe biochemistry-oriented procedures used to find previously obscure flexibility for capsids of the related phages, T3 and T7. The primary procedures are hydration-based buoyant density ultracentrifugation and purified particle-based cryo-electron microscopy (cryo-EM). We review the buoyant density centrifugation in detail. The mature, stable T3/T7 capsid is a shell flexibility-derived conversion product of an initially assembled procapsid (capsid I). During DNA packaging, capsid I expands and loses a scaffolding protein to form capsid II. The following are observations made with capsid II. (1) The in vivo DNA packaging of wild type T3 generates capsid II that has a slight (1.4%), cryo-EM-detected hyper-expansion relative to the mature phage capsid. (2) DNA packaging in some altered conditions generates more extensive hyper-expansion of capsid II, initially detected by hydration-based preparative buoyant density centrifugation in Nycodenz density gradients. (3) Capsid contraction sometimes occurs, e.g., during quantized leakage of DNA from mature T3 capsids without a tail.


[Post suckling prolactin and estradiol levels as predictors of the time of appearance of the first post partum menstruation].
The variability in the duration of lactational amenorrhoea (LA) lead to develop statistical multivariate models to predict the risk of the appearance of the first postpartum menstruation. To estimate the probability of recovering the first postpartum menstruation by means of a survival analysis, including hormonal levels and other parameters as predictor variables. Eighty one mothers in exclusive breastfeeding until the sixth postpartum month, in whom estradiol, basal and post suckling prolactin were measured at the third post partum month, were studied. The variables that better predict the appearance of the first menstruation between the 3rd and 12th postpartum months, were identified using a Cox model survival analysis. The median amenorrhea survival time (the lapse when the chance of recovering menstruation is 50%) was 209 days from delivery. Dichotomized estradiol and post suckling prolactin were the only significant variables that predicted the return of menstruation, with cutoff points of 190 pmol/ and 2,550 mIU/L, respectively. Post suckling prolantin and estradiol levels, measured at the third post partum month, are predictors for the time of appearance of the first postpartum menstruation.


Impulse oscillometry and nitrogen washout test in the assessment of small airway dysfunction in asthma: Correlation with quantitative computed tomography.
Small airway dysfunction (SAD) and airway remodeling influence the disease control and progression in asthma. We investigated whether impulse oscillometry (IOS) and single breath nitrogen washout (SBN2W) could be reliable tests in evaluating SAD and airway remodeling by correlating their data with radiological parameters derived from quantitative chest multidetector computed tomography (MDCT) imaging. Lung function tests were performed before and after bronchodilator. The MDCT lung scans were acquired at full inspiration and expiration using a portable spirometer to control the respiratory manoeuvres. Symptom control was assessed using the Asthma Control Test (ACT) questionnaire. Twenty six patients were enrolled. The bronchial lumen area (LA) measured with MDCT lung scan, correlated inversely with airway resistance (Raw, p < 0.001) and with total and large airway oscillometric resistance (R5, p = 0.002 and R20, p = 0.006, respectively). However these two last correlations became non-significant after Bonferroni correction for multiple comparisons. The radiological quantification of air trapping correlated with Raw (p < 0.001), residual volume (RV, p < 0.001), and the slope of phase III of SBN2W (DeltaN2, p < 0.001) whereas the correlation with small airway oscillometric resistance (R5-20) was non-significant after Bonferroni adjustment. Finally, air trapping was significantly higher in patients with a fixed bronchial obstruction in comparison to patients with reversible obstruction. Plethysmographic method remains the main tool to investigate SAD and airway remodeling in asthmatic patients. The integration with the SBN2W test proved useful to better evaluate the small airway involvement whereas IOS showed a weaker correlation with both radiological and clinical data.


Purification of two distinct proteins of approximate Mr 80,000 from human epithelial cells and identification as proper substrates for protein kinase C.
A Mr-80,000 acidic phosphoprotein ('80K protein') is a specific substrate for protein kinase C. We attempted to purify the 80K protein from a human squamous-cell carcinoma cell line, Ca9-22, by the sequential use of heat treatment, (NH4)2SO4 precipitation, Mono Q column chromatography, proRPC column chromatography and gel filtration. The 80K protein was assayed by phosphorylation in vitro by using partially purified human type III protein kinase C, and was fractionated into two distinct molecular species with slightly different Mr values, designated 80K-L and 80K-H proteins. Phosphorylation occurred mainly at serine residues of these proteins. Two-dimensional phosphopeptide maps after trypsin digestion and kinetic profiles of phosphorylation were different from each other. Ca2(+)- and phospholipid-dependency of the phosphorylation in vitro confirmed that both 80K-L and 80K-H proteins are true substrates for three subtypes of protein kinase C. The 80K-L protein was a preferential substrate for type III protein kinase C, and the 80K-H protein was phosphorylated more effectively by type I and type II protein kinase C. The possible roles of these two distinct 80K proteins in signal transduction are discussed.


Long-Term Limitation Effects of Se(VI), Zn(II), and Ni(II) on Start-Up of the Anammox Process Using Gel Carrier.
Anaerobic ammonia oxidation (anammox) bacteria are inhibited by heavy metals at high concentrations but require trace amounts of some heavy-metal elements for growth and activity maintenance. The present study evaluates the long-term limitation effects of Se(VI), Zn(II), and Ni(II) on the start-up period of an anammox reactor. To strictly limit the levels of heavy metals in the reactor, all tests used ultrapure water as the influent synthetic wastewater and all reactors were installed in a clean booth. The anammox biomass was maintained through the gel entrapment technique. In the absence of Se(VI) and Ni(II), the anammox reactor start-up was 18.9 kg-N (m3-carrier d)-1 (nitrogen conversion rate (NCR) per gel carriers), indicating that Se(VI) and Ni(II) are not required or need not be continuously added to maintain the anammox process. Under Zn(II) limitation, the anammox process failed to start-up and the NCR tended to decrease rapidly. After readdition of 0.005 mg L-1 of Zn(II), the NCR did not decline further and instead partially recovered at a very slow rate. The NCR was completely recovered after adding 0.020 mg L-1 of Zn(II). These results reveal that Zn(II) limitation seriously affects the start-up of the anammox process while Se(VI) and Ni(II) are not required or need not be continuously added to the anammox process.


Absolute and relative risks of kidney and urological complications in patients with inflammatory bowel disease.
The burden of kidney and urological complications in patients with inflammatory bowel disease (IBD) remains poorly characterized. We analyzed association between developing IBD (as a time-varying exposure) and relative risks of receiving diagnoses of chronic kidney disease (CKD), acute kidney injury (AKI), or kidney stones, and experiencing a clinically-relevant decline in estimated glomerular filtration rate (eGFR) (CKD progression; composite of kidney failure or an eGFR decline ≥30%) in 1,682,795 individuals seeking healthcare in Stockholm, Sweden, during 2006-2018. We quantified 5- and 10-year absolute risks of these complications in a parallel matched cohort of IBD cases and random controls matched (1:5) on sex, age, and eGFR. During median 9-years, 10,117 participants developed IBD. Incident IBD was associated with higher risks of kidney-related complications compared with non-IBD periods: HR (95% CI) was 1.24 (1.10-1.40) for receiving a CKD diagnosis, 1.11 (1.00-1.24) for CKD progression. For absolute risks, 11.8% IBD cases had a CKD event within 10-year. Of these, 6.4% received a CKD diagnosis, and 7.9% reached CKD progression. The risks of AKI (HR 1.97 [1.70-2.29]; 10-y absolute risk 3.6%), kidney stones (HR 1.69 [1.48-1.93]; 10-y absolute risk 5.6%) were also elevated. Risks were similar in Crohn's disease and ulcerative colitis. More than 10% of IBD patients developed CKD within 10-year from diagnosis, with many not being identified through diagnostic codes. This, together with their elevated AKI and kidney stone risks, highlights the need of established protocols for kidney function monitoring and referral to nephrological/urological care for IBD patients.


Classification images predict absolute efficiency.
How well do classification images characterize human observers' strategies in perceptual tasks? We show mathematically that from the classification image of a noisy linear observer, it is possible to recover the observer's absolute efficiency. If we could similarly predict human observers' performance from their classification images, this would suggest that the linear model that underlies use of the classification image method is adequate over the small range of stimuli typically encountered in a classification image experiment, and that a classification image captures most important aspects of human observers' performance over this range. In a contrast discrimination task and in a shape discrimination task, we found that observers' absolute efficiencies were generally well predicted by their classification images, although consistently slightly (approximately 13%) higher than predicted. We consider whether a number of plausible nonlinearities can account for the slight under prediction, and of these we find that only a form of phase uncertainty can account for the discrepancy.


Subdural hemorrhage in pediatric patients with enlargement of the subarachnoid spaces.
Enlargement of the subarachnoid spaces has been theorized as a risk factor for the development of subdural hemorrhage (SDH). As the finding of unexplained SDH in children often raises suspicion for nonaccidental trauma, the possibility of increased risk of SDH in children with enlargement of the subarachnoid spaces has important clinical, social, and legal implications. Therefore, the authors evaluated the frequency of SDH in a cohort of children with enlargement of the subarachnoid spaces. The authors identified children younger than 2 years of age who were diagnosed with enlargement of the subarachnoid spaces on MRI or CT scanning in a large primary care network between July 2001 and January 2008. The authors excluded children who had enlargement of the subarachnoid spaces diagnosed on imaging performed for trauma or developmental delay, as well as children with a history of prematurity, diagnosis of intracranial pathology, or metabolic or genetic disorders. Chart review recovered the following data: patient demographics, head circumference, history of head trauma, and head imaging results. For the subset of children with SDH, information regarding evaluation for other injuries, including skeletal survey, ophthalmological examination, and child protection team evaluation, was abstracted. There were 177 children with enlargement of the subarachnoid spaces who met the inclusion criteria. Subdural hemorrhage was identified in 4 (2.3%) of the 177 children. All of the children with SDH underwent evaluations for suspected nonaccidental trauma, which included consultation by the child protection team, skeletal survey, and ophthalmological examination. Additional injuries (healing rib fractures) were identified in 1 of 4 patients. None of the 4 children had retinal hemorrhages. Only the child with rib fractures was reported to child protective services due to concerns for abuse. Only a small minority of the patients with enlargement of the subarachnoid spaces had SDH. Evidence of additional injuries concerning for physical abuse were identified in a quarter of the children with enlargement of the subarachnoid spaces and SDH, suggesting that an evaluation for suspected nonaccidental trauma including occult injury screening should be performed in cases of SDH with enlargement of the subarachnoid spaces. In the absence of additional injuries, however, the presence of an unexplained SDH in the setting of enlargement of the subarachnoid spaces may be insufficient to support a diagnosis of nonaccidental trauma.


Intraocular lens implantation the Nigerian experience.
Between April 1989 and December 1990, 51 patients underwent cataract extractions with implantation of intraocular lenses. A preliminary report of encouraging results is given inspite of limitations of equipment for microsurgical procedures.


[The role of nitric oxide in ethanol postconditioning induced cardioprotection].
To investigate whether the release of nitric oxide (NO) was involved in the cardioprotection of ethanol postconditioning in isolated rat hearts. Hearts isolated from male SD rats were subjected to 30 min of regional ischemia (occlusion of left anterior descending artery) followed by 120 min of reperfusion. Ethanol postconditioning was fulfilled through perfusion of 50 mmol/L ethanol for 15 min (at the end of cardiac ischemia for 5 min and at the beginning of reperfusion for 10 min). The rats were divided into five groups: normal, ischemia and reperfusion, ethanol postconditioning, ethanol postconditioning + L-nitro-arginine-methylester (L-NAME) and ethanol postconditioning + atractyloside. The ventricular hemodynamic parameters and lactate dehydrogenase (LDH) release during reperfusion were measured. The infarct size was measured by TTC staining method and NO content was measured by nitric acid reductase method. The expressions of Bcl-2 and Bax mRNA were detected by RT-PCR analysis. In contrast to ischemia and reperfusion, ethanol postconditioning improved left ventricular developed pressure, rate pressure product during reperfusion, reduced LDH release and infarct size. NO content was decreased. The ratio of Bcl-2/Bax was increased. Administration of nitric oxide synthase inhibitor L-NAME or mitochondrial permeability transition pore opener atractyloside both attenuated the role of ethanol postconditioning, which inhibited the recovery of hemodynamic parameters, the decreases of LDH and infarct size. NO content was decreased further. The ratio of Bcl-2/Bax was decreased. The cardioprotection of ethanol postconditioning may be associated with reducing nitric oxide release, inhibiting the opening of mitochondrial permeability transition pore and decreasing the happening of apoptosis.


Geometry of human vascular system: is it an obstacle for quantifying antiangiogenic therapies?
It is now recognized that all human natural and diseased anatomic systems are characterized by irregular shapes and very complex behaviors. In geometrical terms, tumor vascularity (which is the result of a nonlinear dynamic process called angiogenesis) is an archetypal anatomic system that irregularly fills a 3-dimensional Euclidean space. This characteristic, together with the highly variable nature of vessel shapes and surfaces, leads to considerable spatial and temporal heterogeneity in the delivery of oxygen, nutrients, and drugs, and the removal of metabolites. Although these biologic features have been well established, the quantitative analysis of neovascularity in 2-dimensional histologic sections still fails to view its architecture as a non-Euclidean geometrical object, thus allowing errors in visual interpretation and discordant results concerning the same tumor from different laboratories. We discuss here the tumor-induced vascular system as a fractal object, and what changes this new way of observing may bring to the quantification of effective antiangiogenic therapies.


Expanding the genetic causes of small-fiber neuropathy: SCN genes and beyond.
Small-fiber neuropathy (SFN) is a disorder that exclusively affects the small nerve fibers, sparing the large nerve fibers. Thinly myelinated Aδ-fibers and unmyelinated C-fibers are damaged, leading to development of neuropathic pain, thermal dysfunction, sensory symptoms, and autonomic disturbances. Although many SFNs are secondary and due to immunological causes or metabolic disturbances, the etiology is unknown in up to half of the patients. Over the years, this proportion of "idiopathic SFN" has decreased, as familial and genetic causes have been discovered, thus shifting a proportion of once "idiopathic" cases to the genetic category. After the discovery of SCN9A-gene variants in 2012, SCN10A and SCN11A variants have been found to be pathogenic in SFN. With improved accessibility of SFN diagnostic tools and genetic tests, many non-SCN variants and genetically inherited systemic diseases involving the small nerve fibers have also been described, but only scattered throughout the literature. There are 80 SCN variants described as causing SFN, 8 genes causing hereditary sensory autonomic neuropathies (HSAN) described with pure SFN, and at least 7 genes involved in genetically inherited systemic diseases associated with SFN. This systematic review aims to consolidate and provide an updated overview on the genetic variants of SFN to date---SCN genes and beyond. Awareness of these genetic causes of SFN is imperative for providing treatment directions, prognostication, and management of expectations for patients and their health-care providers.


Study on musculoskeletal complaints involving the back, neck and upper limbs.
A mail questionnaire survey was conducted among designated factory doctors (DFDs) to determine the nature and extent of musculoskeletal aches/pains in patients attending their clinics over one working week. Information was recorded for all patients aged 15 years and above presenting with aches or pains in the back, neck or upper limbs, as the main complaint or as one of the presenting symptoms. For the 155 doctors participating in the survey, the total attendance of patients aged 15 years and above during the study period was 35,010. Of these, 3.9% presented with the symptoms studied and 1.8% had work-related complaints. The commonest site affected was the back (55.7%), followed by the neck (21.4%) and shoulders (19.2%). A higher proportion of males than females had back complaints with the reverse for complaints involving hands/wrists and arms/forearms. 82.3% of the affected were employed, 60.3% being production workers, compared to 33.3% professional/office workers and 6.4% service workers. Of the patients who were working, 51.3% had work-related symptoms and 54.4% were given medical leave. Production workers had the highest proportion with work-related symptoms while service workers had the highest proportion given medical leave. The "medical certificate rate" was highest for back symptoms -57.6%, while work-related symptoms was highest for complaints involving hands/wrists. The study findings are consistent with those of a 1993 morbidity survey of outpatients in Singapore and indicate that the prevalence of work-related musculoskeletal aches/pains is not high.


Biomarkers of oxidative stress study VI. Endogenous plasma antioxidants fail as useful biomarkers of endotoxin-induced oxidative stress.
This is the newest report in a series of publications aiming to identify a blood-based antioxidant biomarker that could serve as an in vivo indicator of oxidative stress. The goal of the study was to test whether acutely exposing Göttingen mini pigs to the endotoxin lipopolysaccharide (LPS) results in a loss of antioxidants from plasma. We set as a criterion that a significant effect should be measured in plasma and seen at both doses and at more than one time point. Animals were injected with two doses of LPS at 2.5 and 5 µg/kg iv. Control plasma was collected from each animal before the LPS injection. After the LPS injection, plasma samples were collected at 2, 16, 48, and 72 h. Compared with the controls at the same time point, statistically significant losses were not found for either dose at multiple time points in any of the following potential markers: ascorbic acid, tocopherols (α, δ, γ), ratios of GSH/GSSG and cysteine/cystine, mixed disulfides, and total antioxidant capacity. However, uric acid, total GSH, and total Cys were significantly increased, probably because LPS had a harmful effect on the liver. The leakage of substances from damaged cells into the plasma may have increased plasma antioxidant concentrations, making changes difficult to interpret. Although this study used a mini-pig animal model of LPS-induced oxidative stress, it confirmed our previous findings in different rat models that measurement of antioxidants in plasma is not useful for the assessment of oxidative damage in vivo.


Disease-Associated Streptococcus pneumoniae Genetic Variation.
Streptococcus pneumoniae is an opportunistic pathogen that causes substantial illness and death among children worldwide. The genetic backgrounds of pneumococci that cause infection versus asymptomatic carriage vary substantially. To determine the evolutionary mechanisms of opportunistic pathogenicity, we conducted a genomic surveillance study in China. We collected 783 S. pneumoniae isolates from infected and asymptomatic children. By using a 2-stage genomewide association study process, we compared genomic differences between infection and carriage isolates to address genomic variation associated with pathogenicity. We identified 8 consensus k-mers associated with adherence, antimicrobial resistance, and immune modulation, which were unevenly distributed in the infection isolates. Classification accuracy of the best k-mer predictor for S. pneumoniae infection was good, giving a simple target for predicting pathogenic isolates. Our findings suggest that S. pneumoniae pathogenicity is complex and multifactorial, and we provide genetic evidence for precise targeted interventions.


Functional neuroimaging in craniopharyngioma: a useful tool to better understand hypothalamic obesity?
To use functional magnetic resonance imaging (fMRI) in craniopharyngioma (CP) patients to examine the hypothesis that hypothalamic damage due to CP and its treatment results in enhanced perception of food reward and/or impaired central satiety processing. Pre- and post-meal responses to visual food cues in brain regions of interest (ROI; bilateral nucleus accumbens, bilateral insula, and medial orbitofrontal cortex) were assessed in 4 CP patients versus 4 age- and weight-matched controls. Stimuli consisted of images of high- ('fattening') and low-calorie ('non-fattening') foods in blocks, alternating with non-food object blocks. After the first fMRI scan, subjects drank a high-calorie test meal to suppress appetite, then completed a second fMRI scan. Within each ROI, we calculated mean z-scores for activation by fattening as compared to non-fattening food images. Following the test meal, controls showed suppression of activation by food cues while CP patients showed trends towards higher activation. These data, albeit in a small group of patients, support our hypothesis that perception of food cues may be altered in hypothalamic obesity (HO), especially after eating, i.e. in the satiated state. The fMRI approach is encouraging for performing future mechanistic studies of the brain response to food cues and satiety in patients with hypothalamic or other forms of childhood obesity.


[Overview of erythrocyte glucose-6-phosphate deficiency].
The cloning and sequencing of the G6PD gene has opened a new chapter in the characterization of the numerous G6PD variants described. Many which were thought to be different on the basis of their kinetic properties, are really the same whereas heterogeneity has been found in some which were thought to be homogeneous. We discuss with some detail variants A+, A- and the Mediterranean type.


A First Insight into the Microbial and Viral Communities of Comau Fjord-A Unique Human-Impacted Ecosystem in Patagonia (42∘ S).
While progress has been made in surveying the oceans to understand microbial and viral communities, the coastal ocean and, specifically, estuarine waters, where the effects of anthropogenic activity are greatest, remain partially understudied. The coastal waters of Northern Patagonia are of interest since this region experiences high-density salmon farming as well as other disturbances such as maritime transport of humans and cargo. Here, we hypothesized that viral and microbial communities from the Comau Fjord would be distinct from those collected in global surveys yet would have the distinctive features of microbes from coastal and temperate regions. We further hypothesized that microbial communities will be functionally enriched in antibiotic resistance genes (ARGs) in general and in those related to salmon farming in particular. Here, the analysis of metagenomes and viromes obtained for three surface water sites showed that the structure of the microbial communities was distinct in comparison to global surveys such as the Tara Ocean, though their composition converges with that of cosmopolitan marine microbes belonging to Proteobacteria, Bacteroidetes, and Actinobacteria. Similarly, viral communities were also divergent in structure and composition but matched known viral members from North America and the southern oceans. Microbial communities were functionally enriched in ARGs dominated by beta-lactams and tetracyclines, bacitracin, and the group macrolide-lincosamide-streptogramin (MLS) but were not different from other communities from the South Atlantic, South Pacific, and Southern Oceans. Similarly, viral communities were characterized by exhibiting protein clusters similar to those described globally (Tara Oceans Virome); however, Comau Fjord viromes displayed up to 50% uniqueness in their protein content. Altogether, our results indicate that microbial and viral communities from the Comau Fjord are a reservoir of untapped diversity and that, given the increasing anthropogenic impacts in the region, they warrant further study, specifically regarding resilience and resistance against antimicrobials and hydrocarbons.


Tension transients initiated by photogeneration of MgADP in skinned skeletal muscle fibers.
Addition of MgADP to skinned skeletal muscle fibers causes a rise in Ca(2+)-activated isometric tension. Mechanisms underlying this tension increase have been investigated by rapid photogeneration of ADP within skinned single fibers of rabbit psoas muscle. Photolysis of caged ADP (P2-1(2-nitrophenyl)ethyladenosine 5'-diphosphate) resulted in an exponential increase in isometric tension with an apparent rate constant, kADP, of 9.6 +/- 0.3 s-1 (mean +/- SE, n = 28) and an amplitude, PADP, of 4.9 +/- 0.3% Po under standard conditions (0.5 mM photoreleased MgADP, 4 mM MgATP, pH 7.0, pCa 4.5, 0.18 M ionic strength, 15 degrees C). PADP depended upon the concentration of photoreleased MgADP as well as the concentration of MgATP. A plot of 1/PADP vs. 1/[MgADP] at three MgATP concentrations was consistent with competition between MgADP and MgATP for the same site on the crossbridge. The rate of the transient, kADP, also depended upon the concentration of MgADP and MgATP. At both 4 and 1 mM MgATP, kADP was not significantly different after photorelease of 0.1-0.5 mM MgADP, but was reduced by 28-40% when 3.5 mM MgADP was added before photorelease of 0.5 mM MgADP. kADP was accelerated by about twofold when MgATP was varied from 0.5 to 8 mM MgATP. These effects of MgATP and MgADP were not readily accounted for by population of high force-producing states resulting from reversal of the ADP dissociation process. Rather, the results suggest that competition between MgADP and MgATP for crossbridges at the end of the cycle slows detachment leading to accumulation of force-generating crossbridges. Elevation of steady-state Pi concentration from 0.5 to 30 mM caused acceleration of kADP from 10.2 +/- 0.5 to 27.8 +/- 1.8 s-1, indicating that the tension rise involved crossbridge flux through the Pi dissociation step of the cycle.


The Virtual-Spine Platform-Acquiring, visualizing, and analyzing individual sitting behavior.
Back pain is a serious medical problem especially for those people sitting over long periods during their daily work. Here we present a system to help users monitoring and examining their sitting behavior. The Virtual-Spine Platform (VSP) is an integrated system consisting of a real-time body position monitoring module and a data visualization module to provide individualized, immediate, and accurate sitting behavior support. It provides a comprehensive spine movement analysis as well as accumulated data visualization to demonstrate behavior patterns within a certain period. The two modules are discussed in detail focusing on the design of the VSP system with adequate capacity for continuous monitoring and a web-based interactive data analysis method to visualize and compare the sitting behavior of different persons. The data was collected in an experiment with a small group of subjects. Using this method, the behavior of five subjects was evaluated over a working day, enabling inferences and suggestions for sitting improvements. The results from the accumulated data module were used to elucidate the basic function of body position recognition of the VSP. Finally, an expert user study was conducted to evaluate VSP and support future developments.


Psychometric Testing of the Control Attitudes Scale-Revised for Patients With a Left Ventricular Assist Device.
Perceived control, an indicator of the patient's ability to cope and thrive with a chronic illness, is a common target of nursing interventions. As of 2019, more than 25 000 patients had been implanted with a left ventricular assist device (LVAD) as a treatment of advanced heart failure. Patients with an LVAD experience significant life changes that affect anxiety, depression, health-related quality of life, and, presumably, perceived control. To adequately intervene and improve perceived control, a reliable and valid measure is needed. The objectives of this analysis were to (1) assess item discrimination and anticipated range of scores of the Control Attitudes Scale-Revised (CAS-R), (2) assess the internal consistency and validity of the CAS-R, and (3) examine perceived control in a sample of patients with an LVAD. Two cohorts of patients with an LVAD (n = 113) were combined to evaluate the psychometric qualities of the CAS-R. Correlations among patient-reported outcomes and perceived control were used to evaluate validity. Cronbach α was used to test internal consistency. Item response theory was used to measure item discrimination and anticipated scores. Descriptive statistics describe perceived control in the sample. Overall, the CAS-R demonstrated good internal consistency and convergent validity with other patient-reported outcomes. Using the item response theory, we saw that the CAS-R was a good predictor of lower-moderate scorers but was not good at differentiating high performers. There were several items that were poor discriminators and could be altered or discarded to create a more predictive instrument. The CAS-R is a valid and reliable instrument to measure perceived control in patients who have LVAD implants; however, more work could be done to improve item-level information.


Expression of cell surface antigens by suppressor T cell hybridomas. I. Comparison of phenotype and function.
The phenotypic expression of cell surface markers by T cell hybridomas that elaborate suppressor factors specific for the polymers L-glutamic acid60-L-alanine30-L-tyrosine10 (GAT) or L-glutamic acid50-L-tyrosine50 (GT) has been analyzed. We found that determinants encoded by the I-J subregion of the H-2 complex were borne on the surface of these hybrid cells and on the factors they secrete, whereas I-J determinants were not expressed by the AKR thymoma fusion parent, BW5147. The level of expression of I-J determinants fluctuated widely depending upon culture conditions, but I-J products and other cell surface markers of normal T cells could be quantitatively increased, or induced to appear, by treatment of the hybridomas with chemical agents, such as dimethyl sulfoxide (DMSO) or phorbol myristate acetate (PMA). In contrast, the surface expression of the viral product gp70 was decreased by the same treatment. Using chemical induction, we typed BW5147, a group of antigen-specific suppressor T cell hybridomas, and two control hybridomas for expression of I-J, Thy-1, Lyt, and H-2K alloantigens. Also, a haplotype-specific hybridoma that produces an antigen-nonspecific factor was analyzed. The results demonstrated that BW5147 failed to express I-J or Lyt alloantigens but expressed Thy-1.1 and H-2Kk gene products. The pattern of expression of these antigens by T cell hybridomas was very complex, but three conclusions could be drawn: 1) Good correlation exists between the expression of certain I-J determinants and the ability of T cell hybridomas to produce suppressor factor. 2) The expression of Thy-1, Lyt, or H-2Kk determinants is variable, and no correlation was found between expression of these antigens and the ability to produce active suppressor factors. 3) I-Jk products contributed by the AKR thymoma fusion partner are expressed by T cell hybridomas.


Mutagenicity and pausing of HIV reverse transcriptase during HIV plus-strand DNA synthesis.
The unusually high frequency of misincorporation by HIV-1 reverse transcriptase (HIV RT) is likely to be the major factor in the rapid accumulation of viral mutations in AIDS, especially in the env gene. To investigate the ability of HIV RT to copy the env gene, we subcloned an HIV env gene fragment into a single-stranded DNA vector and measured the progression of synthesis by HIV RT. We observed that HIV RT, but not RT from avian myeloblastosis virus, DNA polymerase-alpha or T7 DNA polymerase, pauses specifically at poly-deoxyadenosine stretches within the env gene. The frequency of bypassing the polyadenosine stretches by HIV RT is enhanced by increasing the ratio of enzyme to template. We measured the fidelity of DNA synthesis within a segment of the hypervariable region 1 of the env gene (V-1) containing a poly-deoxyadenosine sequence by repetitively copying the DNA by HIV RT, and then cloning and sequencing the copied fragments. We found that 27% of the errors identified in V-1 sequence were frameshift mutations opposite the poly-adenosine tract, a site where strong pausing was observed. Pausing of HIV RT at the polyadenosine tract could be enhanced by either distamycin A or netropsin, (A-T)-rich minor groove binding peptides. Moreover, netropsin increases the frequency of frameshift mutations in experiments in which HIV RT catalyzes gap filling synthesis within the lacZ gene in double-stranded circular M13mp2 DNA. These combined results suggest that the enhanced mutation frequency may be due to increased pausing at netropsin-modified polyadenosine tracts. Therefore, netropsin and related A-T binding chemicals may selectively enhance frameshift mutagenesis induced by HIV RT and yield predominantly non-viable virus.


Pharmacokinetic and pharmacodynamic activities of ciprofloxacin against strains of Streptococcus pneumoniae, Staphylococcus aureus, and Pseudomonas aeruginosa for which MICs are similar.
The serum bactericidal activity of ciprofloxacin against strains of Streptococcus pneumoniae, Staphylococcus aureus, and Pseudomonas aeruginosa for which MICs are similar (0.4 microgram/ml) was assessed with serum ultrafiltrates from five healthy volunteers receiving ciprofloxacin at 400 mg intravenously every 8 h. In addition, human serum was supplemented with ciprofloxacin to achieve a mean steady-state concentration (Css) that might be achieved in patients with renal failure, with total clearances of 3 to 4 liters/h (elimination rate constant, 0.08 h-1). The area under the inhibitory titer curve from 0 to 24 h (AUIC24) and the area under the bactericidal titer curve from 0 to 24 h (AUBC24) were both measured and predicted as the area under the concentration-time curve from 0 to 24 h (AUC24)/MIC and AUC24/MBC, respectively. We previously demonstrated that a breakpoint AUC24/MIC of 125 for ciprofloxacin had a significantly higher probability of treatment success than lower values, with 250 to 500 being optimal. Volunteer sera (mean Css, 1.55 to 2.48 micrograms/ml) achieved AUC24/MICs of 90 to 145. Supplemented serum (mean Css, 6.00 to 7.42 micrograms/ml) achieved AUC24/MICs of 350 to 450. Correlation coefficients for measured and predicted values of AUC24/MIC and AUC24/MBC were 0.826 and 0.941, respectively. The mean percent errors were not significantly different from zero for either AUIC24 or AUBC24 values (P > 0.1, P > 0.4). Time-kill curve studies were performed with low (1.55 to 2.48 micrograms/ml), intermediate (6.00 to 7.42 micrograms/ml), and high (15 to 25 micrograms/ml) concentrations of ciprofloxacin for the three organisms. At low concentrations (3 to 6 times the MIC) AUC24/MICs were <125 for two of five volunteers and the killing rates were considerably more rapid for P. aeruginosa than for S. pneumoniae or S.aureus. Intermediate concentrations (15 to 18 times the MIC) achieved optimal AUC24/MICs, and the killing rates were similar for the three organisms. A paradoxical decrease in the killing rate was seen at high concentrations (35 to 60 times the MIC). At clinically achievable concentrations, ciprofloxacin killed P. aeruginosa more rapidly than it did either S. pneumoniae or S. aureus.


Serum or cerebrospinal fluid levels of glyceraldehyde-derived advanced glycation end products (AGEs) may be a promising biomarker for early detection of Alzheimer's disease.
Alzheimer's disease (AD) is the most common cause of dementia in developed countries. AD is characterized pathologically by the presence of senile plaques and neurofibrillary tangles (NFTs), the major constituents of which are the amyloid beta protein (Abeta) and tau protein, respectively. Several epidemiological studies have reported moderately increased risks of AD in diabetic patients compared with general population. In diabetes mellitus, the formation and accumulation of advanced glycation end products (AGEs) progress. Recent understandings of this process have confirmed that AGEs - their receptor (RAGE) interactions may play a role in the pathogenesis of diabetic vascular complications and neurodegenerative disorders including AD. Indeed, it has been demonstrated that AGEs can be identified immunohistochemically to be present in both senile plaques and NFTs from patients with AD. Glycation of Abeta markedly enhances its aggregation in vitro, and the glycation of tau, in addition to hyperphosphorylation, appears to enhance the formation of paired helical filaments. Further, RAGE has been found a specific cell surface receptor for Abeta peptite, thus eliciting neuronal cell perturbation. The active participation of RAGE in the pathogenesis of AD has also been confirmed in RAGE-overexpressed transgenic mice. Moreover, we have recently found that glyceraldehyde-derived AGEs, one of the representative ligands for RAGE, exerted cytopathic effects on cultured neuronal cells and that neurotoxic effect of diabetic serum was completely blocked by neutralizing antibodies against glyceraldehydes-derived AGEs. These observations led us to hypothesize that serum or cerebrospinal fluid (CSF) levels of glyceraldehyde-derived AGEs could become a promising biomarker for early detection of AD. We also would like to propose the possible ways of testing our hypothesis. Are the concentrations of glyceraldehyde-derived AGEs in serum or CSF elevated early in the course of dementia? Are these levels correlated with disease severity and progression, especially in patients with diabetes? These clinical studies clarify whether use of serum or CSF levels of glyceraldehyde-derived AGEs as a biomarker for AD might enable more effective diagnosis and treatment of patients with this devastating disorder.


Sub-chronic exposure to arsenic and dichlorvos on erythrocyte antioxidant defense systems and lipid peroxidation in rats.
The effect of combined exposure to arsenic (25 ppm in drinking water) and dichlorvos (2.5 mg kg1, orally) for 56 days on biochemical variables, indicative of lipid peroxidation, antioxidant enzyme system and AChE activity in erythrocytes of rats, were examined. While arsenic caused a significant increase in AChE, DDVP produced marked depletion. Combined exposure to arsenic and DDVP produced no additional decrease in AChE activity, which was comparable to DDVP. Arsenic and DDVP also increased the levels of reactive oxygen species (ROS) and thiobarbituric acid reactive substances (TBARS), suggesting free radical generation. Interestingly, glutathione linked enzymes (GSH, GPx, GST and GR) significantly increased on arsenic and DDVP exposure. SOD activity also increased significantly in the individually exposed groups, while catalase activity remained unchanged. Blood arsenic level increased significantly on coexposure to arsenic alone and with DDVP exposed group. However, arsenic content in co-exposed group depleted marginally as compared to arsenic alone group, indicating possible arsenic redistribution. It might be concluded from the study that the combined exposure to arsenic and DDVP may lead to synergistic effects on certain biochemical indicators of oxidative stress like ROS, GSH and SOD, suggesting a more pronounced induction of lipid peroxidation in erythrocytes.


Radial Head and Neck Allograft for Comminute Irreparable Fracture-Dislocations of the Elbow.
Fracture-dislocations of the elbow can be difficult to treat, with unsatisfactory results in some cases. In general, it is preferable to preserve the fractured radial head when possible, but some patients present a unique treatment challenge because of extremely comminuted fractures and bone loss. In these cases, the only options available are radial head prosthesis or allograft. The authors present a case of a 45-year-old man with a fracture-dislocation of the left elbow that was treated with an allograft of the radial head and neck because of extreme comminution of the fracture. There have been a few reports about osteochondral allograft transplantation of the radial head, and they all included traumatic or posttraumatic cases treated with a frozen allograft. To the best of the authors' knowledge, this is the first report on the use of osteochondral allograft in the acute setting for the treatment of a comminuted fracture of the radius involving the whole head and neck. The clinical results were satisfactory at the final follow-up, although mild degenerative changes were present, the screws were coming loose, and the radial head had a slight valgus deformity. Radial head allograft can be an option in selected cases of acute fractures with severe comminution and bone loss that are not amenable to a stable internal fixation; for the young and active patient, who is not the best candidate for radial head resection; or in cases in which radial head arthroplasty is not feasible because of severe bone loss. [Orthopedics. 2016; 39(6):e1205-e1208.].


DUSP23 is over-expressed and linked to the expression of DNMTs in CD4+ T cells from systemic lupus erythematosus patients.
We evaluated the transcriptional expression of dual-specificity protein phosphatase 23 (DUSP23) in CD4+ T cells from 30 systemic lupus erythematosus (SLE) patients and 30 healthy controls. DUSP23 mRNA levels were considerably higher in the patient group: 1490 ± 1713 versus 294·1 ± 204·2. No association was found between DUSP23 mRNA expression and the presence of typical serological and clinical parameters associated with SLE. Meaningful statistical values were obtained in the patient group between the levels of DUSP23 and integrin subunit alpha L (ITGAL), perforin 1 (PRF1) and CD40L. Similarly, transcript levels of different DNA methylation-related enzymes [DNA methylation-related enzymes (DNMT1, DNMT3A, DNMT3B, MBD2, and MBD4)] were also correlated positively with the expression of DUSP23. In an attempt to counteract the hypomethylation status of the promoters of certain genes known to be over-expressed in SLE, it is possible that DUSP23 acts as a negative regulatory mechanism which ultimately silences the transcription of these epigenetically regulated genes by triggering an increase in the expression of different DNMTs.


Combined treatment with the histone deacetylase inhibitor LBH589 and a splice-switch antisense oligonucleotide enhances SMN2 splicing and SMN expression in Spinal Muscular Atrophy cells.
Spinal muscular atrophy (SMA) is a motor neuron disease caused by loss of function mutations in the Survival Motor Neuron 1 (SMN1) gene and reduced expression of the SMN protein, leading to spinal motor neuron death, muscle weakness and atrophy. Although humans harbour the highly homologous SMN2 gene, its defective splicing regulation yields a truncated and unstable SMN protein. The first therapy for SMA was recently approved by the Food and Drug Administration and consists of an antisense oligonucleotide (Nusinersen) rendering SMN2 functional and thus improving patients' motor activity and quality of life. Nevertheless, not all patients equally respond to this therapy and the long-term tolerability and safety of Nusinersen are still unknown. Herein, in vivo splicing assays indicated that the HDAC inhibitor LBH589 is particularly efficient in rescuing the SMN2 splicing defect in SMA fibroblasts and SMA type-I mice-derived neural stem cells. Western blot analyses showed that LBH589 also causes a significant increase in SMN protein expression in SMA cells. Moreover chromatin immunoprecipitation analyses revealed that LBH589 treatment induces widespread H4 acetylation of the entire SMN2 locus and selectively favors the inclusion of the disease-linked exon 7 in SMN2 mature mRNA. The combined treatment of SMA cells with sub-optimal doses of LBH589 and of an antisense oligonucleotide that mimic Nusinersen (ASO_ISSN1) elicits additive effects on SMN2 splicing and SMN protein expression. These findings suggest that HDAC inhibitors can potentiate the activity of Nusinersen and support the notion that 'SMN-plus' combinatorial therapeutic approaches might represent an enhanced opportunity in the scenario of SMA therapy.


Current challenges facing the determination of product bioequivalence in veterinary medicine.
Despite the pharmacological and statistical advances that have occurred since the early days of bioequivalence assessments, there remain many unresolved issues associated with the bioequivalence evaluation of human and veterinary pharmaceuticals. While many of these issues are common to both human and veterinary medicine, there are also challenges specific to veterinary drug products. Examples of complex problems that remain to be resolved include the assessment of drugs associated with complex kinetics (e.g., sustained release formulations that produce multiple peaks), the evaluation of intramammary formulations, uncertainty associated with conditions under which specific enantiomers of metabolites need to be factored into the bioequivalence evaluation, the study design for products and active pharmaceutical ingredients that exhibit highly variable kinetics, equivalence of biomass products, methods for evaluating topical formulations or formulations with very long duration of release, the evaluation of products where destructive sampling is necessary (e.g., aquaculture products), and the evaluation of bioequivalence for Type A medicated articles. This manuscript highlights many of the unresolved challenges currently impacting the evaluation of product bioequivalence in veterinary medicine, and provides a summary of the associated scientific complexities with each of these issues.


Why Do Some Sex Chromosomes Degenerate More Slowly Than Others? The Odd Case of Ratite Sex Chromosomes.
The hallmark of sex chromosome evolution is the progressive suppression of recombination which leads to subsequent degeneration of the non-recombining chromosome. In birds, species belonging to the two major clades, Palaeognathae (including tinamous and flightless ratites) and Neognathae (all remaining birds), show distinctive patterns of sex chromosome degeneration. Birds are female heterogametic, in which females have a Z and a W chromosome. In Neognathae, the highly-degenerated W chromosome seems to have followed the expected trajectory of sex chromosome evolution. In contrast, among Palaeognathae, sex chromosomes of ratite birds are largely recombining. The underlying reason for maintenance of recombination between sex chromosomes in ratites is not clear. Degeneration of the W chromosome might have halted or slowed down due to a multitude of reasons ranging from selective processes, such as a less pronounced effect of sexually antagonistic selection, to neutral processes, such as a slower rate of molecular evolution in ratites. The production of genome assemblies and gene expression data for species of Palaeognathae has made it possible, during recent years, to have a closer look at their sex chromosome evolution. Here, we critically evaluate the understanding of the maintenance of recombination in ratites in light of the current data. We conclude by highlighting certain aspects of sex chromosome evolution in ratites that require further research and can potentially increase power for the inference of the unique history of sex chromosome evolution in this lineage of birds.


Space-group and origin ambiguity in macromolecular structures with pseudo-symmetry and its treatment with the program Zanuda.
The presence of pseudo-symmetry in a macromolecular crystal and its interplay with twinning may lead to an incorrect space-group (SG) assignment. Moreover, if the pseudo-symmetry is very close to an exact crystallographic symmetry, the structure can be solved and partially refined in the wrong SG. Typically, in such incorrectly determined structures all or some of the pseudo-symmetry operations are, in effect, taken for crystallographic symmetry operations and vice versa. A mistake only becomes apparent when the R(free) ceases to decrease below 0.39 and further model rebuilding and refinement cannot improve the refinement statistics. If pseudo-symmetry includes pseudo-translation, the uncertainty in SG assignment may be associated with an incorrect choice of origin, as demonstrated by the series of examples provided here. The program Zanuda presented in this article was developed for the automation of SG validation. Zanuda runs a series of refinements in SGs compatible with the observed unit-cell parameters and chooses the model with the highest symmetry SG from a subset of models that have the best refinement statistics.


Tracking progress on health and climate change in Europe.
Left unabated, climate change will have catastrophic effects on the health of present and future generations. Such effects are already seen in Europe, through more frequent and severe extreme weather events, alterations to water and food systems, and changes in the environmental suitability for infectious diseases. As one of the largest current and historical contributors to greenhouse gases and the largest provider of financing for climate change mitigation and adaptation, Europe's response is crucial, for both human health and the planet. To ensure that health and wellbeing are protected in this response it is essential to build the capacity to understand, monitor, and quantify health impacts of climate change and the health co-benefits of accelerated action. Responding to this need, the Lancet Countdown in Europe is established as a transdisciplinary research collaboration for monitoring progress on health and climate change in Europe. With the wealth of data and academic expertise available in Europe, the collaboration will develop region-specific indicators to address the main challenges and opportunities of Europe's response to climate change for health. The indicators produced by the collaboration will provide information to health and climate policy decision making, and will also contribute to the European Observatory on Climate and Health.


Mortality Effects of Hypothetical Interventions on Physical Activity and TV Viewing.
Long-term effects of physical activity and television (TV) viewing on mortality have been inferred from observational studies. The associations observed do not allow for inferences about the effects of population interventions and could be subject to bias due to time-varying confounding. Using data from the Australian Diabetes, Obesity and Lifestyle Study, collected in 1999-2000 (T0), 2004-2005 (T1), and 2011-2012 (T2), we applied the parametric g-formula to estimate cumulative risks of death under hypothetical interventions on physical activity and/or TV viewing determined from self-report while adjusting for time-varying confounding. In the 6377 participants followed up for 13 yr from 2004 to 2005 to death or censoring in 2017, 781 participants died. The observed cumulative risk of death was 12.2%. The most effective hypothetical intervention was to increase weekly physical activity to >300 min (risk ratio (RR), 0.66 (0.46-0.86) compared with a "worst-case" scenario; RR, 0.83 (0.73-0.94) compared with no intervention). Reducing daily TV viewing to <2 h in addition to physical activity interventions did not show added survival benefits. Reducing TV viewing alone was least effective in reducing mortality (RR, 0.85 (0.60-1.10) compared with the worst-case scenario; RR, 1.06 (0.93-1.20) compared with no intervention). Our findings suggested that sustained interventions to increase physical activity could lower all-cause mortality over a 13-yr period, and there might be limited gain from intervening to reduce TV viewing time in a relatively healthy population.


Validation of a computerized use of time recall for activity measurement in advanced-aged adults.
The Multimedia Activity Recall for Children and Adults (MARCA) is a computerized recall instrument that records use of time during 24 hr the previous day and has been developed to address limitations of current self-report physical activity measures for those in advanced age. Test-retest reliability and convergent validity of the adult MARCA were assessed in a sample of 45 advanced-age adults (age 84.9 SD ± 1.62 yr) as a subsample of the Life and Living in Advanced-Age Cohort Study New Zealand (LiLACS NZ). Test-retest methods required participants to recall the previous day's activity using the MARCA twice within the same day. Convergent validity was assessed against accelerometry. Test-retest reliability was high, with ICCs greater than .99 for moderate to vigorous physical activity (MVPA) and physical activity level (PAL). Compared with accelerometry, the MARCA demonstrated validity comparable to other self-report instruments with Spearman's coefficients of .34 and .59 for time spent in nonsedentary physical activity and PAL. The MARCA is a valid and reliable self-report tool for physical activity behaviors in advanced-age adults.


Benchmark thermochemistry of chloramines, bromamines, and bromochloramines: halogen oxidants stabilized by electron correlation.
Chloramines, bromamines, and bromochloramines are halogen-containing oxidants that arise from the reaction of hypohalous acids with ammonia in water. Although relevant to both water disinfection chemistry and biochemistry, these molecules are difficult to study in the laboratory, and their thermochemical properties remain poorly established. We developed a benchmark level ab initio calculation protocol, termed TA14, adapted from the Weizmann theory and Feller-Peterson-Dixon approaches to determine the molecular structures and thermochemical properties of these compounds. We find that the halamine molecules are bound largely, and in some cases entirely, by electron correlation forces. This presumably explains their high reactivity as electrophilic oxidants. We provide computed heats of formation at 0 K (ΔfH(0)(0 K)) and at 298 K (ΔfH(0)(298 K)) and Gibbs free energies of formation at 298 K (ΔfG(0)(298 K)) for the 9 inorganic chloramines, bromamines, bromochloramines in gas phase. Based on comparisons to previous theoretical and experimental data for a set of 11 small molecules containing N, O, H, Cl, and Br, we propose uncertainties ranging from 1 to 3 kJ mol(-1) for computed thermodynamic properties of the halamines. Reported thermochemical data enable the determination of equilibrium constants for reactions involving halamines, opening possibilities for more quantitative studies of the chemistry of these poorly understood compounds.


Intraarrest rhythms and rhythm conversion in asphyxial cardiac arrest.
The objective was to analyze the cardiac arrest rhythms presenting during asphyxial cardiac arrest (ACA). Asphyxial cardiac arrest was induced in 30 Landrace large white piglets, aged 12 to 15 weeks and with a mean (±SD) weight of 20 (±2) kg. After the onset of cardiac arrest, the animals were left untreated for 4 minutes, after which cardiopulmonary resuscitation was commenced. Heart rhythms were monitored from the onset of asphyxia until return of spontaneous circulation or death. After endotracheal tube clamping and prior to cardiac arrest, normal sinus rhythm was noted in 14 animals, atrial fibrillation in two animals, Mobitz II atrioventricular block in 10 animals, and third-degree atrioventricular block in four animals. At the onset of cardiac arrest, seven animals had ventricular fibrillation (VF), two had asystole, and 21 had pulseless electrical activity (PEA). During the 4-minute period of untreated arrest, however, significant changes in the monitored rhythm were noted; at the end of the fourth minute, 19 animals had VF, two animals had asystole, and nine animals had PEA. The most common rhythm after 4 minutes of untreated ACA was VF, while in 57% of animals, PEA was spontaneously converted to VF during the cardiac arrest interval.


Impact of the Hole Transport Layer on the Charge Extraction of Ruddlesden-Popper Perovskite Solar Cells.
Recent works demonstrate that polyelectrolytes as a hole transport layer (HTL) offers superior performance in Ruddlesden-Popper perovskite solar cells (RPPSCs) compared to poly(3,4-ethylenedioxythiophene) polystyrene sulfonate (PEDOT:PSS). The factors contributing to such improvement need to be systematically investigated. To achieve this, we have systematically investigated how the two HTLs affect the morphology, crystallinity, and orientation of the Ruddlesden-Popper perovskite (RPP) films as well as the charge extraction of the RPPSCs. PEDOT:PSS as a HTL leads to RPP films of low crystallinity and with a number of large pinholes. These factors lead to poor charge carrier extraction and significant charge recombination in the RPPSCs. Conversely, a PCP-Na HTL gives rise to highly crystalline and pinhole-free RPPSC films. Moreover, a PCP-Na HTL provides a better energy alignment at the perovskite/HTL interface because of its higher work function compared to PEDOT:PSS. Consequently, devices using PCP-Na as HTLs are more efficient in extracting charge carriers.


Layered double hydroxide nanoparticles as cellular delivery vectors of supercoiled plasmid DNA.
We prepared stable homogeneous suspensions with layered double hydroxide (LDH) nanoparticles for in vitro gene delivery tests. The viability of HEK 293T cells in the presence of LDH nanoparticles at different concentrations was investigated. This revealed 50% cell viability at 500 microg/mL of LDH nanoparticles that is much higher than 50-100 microg/mL used for the delivery tests. The supercoiled pEF-eGFP plasmid (ca. 6100 base pairs) was mixed with LDH nanoparticle suspensions for anion exchange at a weight ratio of DNA/LDH between 1:25 and 1:100. In vitro experiments show that GFP expression in HEK 293T cells starts in the first day, reaches the maximum levels by the second day and continues in the third day. The GFP expression generally increases with the increase in DNA loading in DNA-LDH nanohybrids. However, the delivery efficiency with LDH nanoparticles as the agent is low. For example, the relative efficiency is 7%-15% of that of the commercial agent FuGENE 6. Three to 6% of total cells expressed GFP in an amount detectable by the FACS cytometry 2 days after transfection at 1 microg/mL of plasmid DNA with 25 microg/mL of LDH nanomaterial. The lower delivery efficiency could be attributed to the aggregation of LDH nanoparticles caused by the long-chain plasmid DNA.


Association of Hearing Loss with Physical, Social, and Mental Activity Engagement.
This article aims to evaluate a hearing loss intervention versus an aging education intervention on activity engagement in the Aging and Cognitive Health Evaluation in Elders pilot (ACHIEVE-P). Forty adults (70-84 years) with hearing loss recruited from the Atherosclerosis Risk in Communities Study and de novo participated. Participants were randomized 1:1 to a best practices hearing intervention or a successful aging intervention. Hearing was measured with pure-tone audiometry. The Community Healthy Activities Model Program for Seniors questionnaire measured self-reported time engaging in activities at baseline and 6-month follow-up. At baseline, greater hearing loss was associated with reduced time per week on mental activities (-3.0 hours per 10 dB of hearing loss, 95% confidence interval: -5.8, -0.2). Mental activity engagement increased (mean: +1.3 hours, SD = 6.6) for the hearing intervention group but decreased (mean: -1.1 hours, SD = 4.8) for the aging education group (Cohen's d : 0.41). Hearing loss may be associated with reduced engagement in mental activities. Whether hearing loss treatment impacts activity will be studied in the full-scale ACHIEVE trial.


MCL-1 is a prognostic indicator and drug target in breast cancer.
Analysis of publicly available genomic and gene expression data demonstrates that MCL1 expression is frequently elevated in breast cancer. Distinct from other pro-survival Bcl-2 family members, the short half-life of MCL-1 protein led us to investigate MCL-1 protein expression in a breast cancer tissue microarray and correlate this with clinical data. Here, we report associations between high MCL-1 and poor prognosis in specific subtypes of breast cancer including triple-negative breast cancer, an aggressive form that lacks targeted treatment options. Deletion of MCL-1 in the mammary epithelium of genetically engineered mice revealed an absolute requirement for MCL-1 in breast tumorigenesis. The clinical applicability of these findings was tested through a combination of approaches including knock-down or inhibition of MCL-1 to show triple-negative breast cancer cell line dependence on MCL-1 in vitro and in vivo. Our data demonstrate that high MCL-1 protein expression is associated with poor outcome in breast cancer and support the therapeutic targeting of MCL-1 in this disease.


Dilution and non-Fermi-liquid effects in the CePtIn Kondo lattice.
Measurements of electrical resistivity (ρ(T)), magnetoresistivity (MR), magnetic susceptibility (χ(T)) and heat capacity (C(P)(T)) are presented for the (Ce(1-x)La(x))PtIn alloy system of which the CePtIn parent is a known dense Kondo compound that does not order magnetically down to 50 mK. χ(T) for alloys 0≤x≤0.8 exhibits Curie-Weiss behaviour. ρ(T) results indicate a transition from a dense Kondo behaviour for 0≤x≤0.2 to a single-ion Kondo region (0.3≤x≤0.8). The Kondo energy scale as given by T(K) values calculated from MR studies and by the temperature T(max)(ρ(mag)) where the magnetic contribution to ρ(T) exhibits a maximum value, is compared with theoretical models. It is shown that the experimental results not only depend on a volume effect as given by the compressible Kondo lattice model of Lavagna but in addition confirm the more complex behaviour recently presented by Burdin and Fulde for a Kondo alloy system in which the magnetic (Ce) and non-magnetic (La) atoms are distributed randomly. Non-Fermi-liquid behaviour is predicted by Burdin and Fulde at certain critical concentrations of the alloy system and experimental evidence for this is presented through χ(T), ρ(T) and C(P)(T) measurements.


What demographers can learn from fruit fly actuarial models and biology.
Historically demographers have viewed the results of actuarial studies of nonhuman species, particularly those on invertebrates such as fruit flies, as largely irrelevant to investigations on human populations. In this paper I present life table data from large scale studies on the Mediterranean fruit fly, and show that they provide important insights into fundamental aspects of mortality relevant to human populations: the trajectory of mortality at older ages, sex mortality differentials, the concept of maximal life span, and demographic heterogeneity and selection. An overriding theme of the paper is the need for demographers to acquire a heightened awareness of new developments in biology including areas such as evolutionary ecology, experimental demography, and molecular medicine.


Genome-wide overexpression screen for activators of antioxidant gene transcription.
Genome-wide screens have proven powerful in associating gene products with certain phenotypes or signal transduction pathways, and thus are valuable tools to ascribe gene function. These genomic screens can be extended to discover genes/proteins that attenuate oxidative stress-induced damage, which is implicated in aging, neurodegenerative disorders, and other diseases. One mechanism by which humans protect themselves from oxidative stress is through an endogenous stress response that leads to transcriptional activation of the antioxidant response element (ARE). The ARE is located in the 59-flanking regions of many phase II detoxification and antioxidant enzymes such as NAD(P)H:quinone oxidoreductase 1 (NQO1) and regulates the expression of these genes. Increasing the levels of antioxidant enzymes without causing oxidative stress can potentially counteract degeneration and may be therapeutically useful. On a genomic scale, ARE activators can be identified by screening expression cDNA libraries in a high-throughput amenable reporter gene assay. Further validation of putative hits requires testing of cDNAs for their ability to upregulate the expression of endogenous ARE-regulated genes on the transcript and protein levels, and their ability to protect cells from oxidative insults. General screening procedure and subsequent hit validation are discussed in detail.


Determining the general circulation of the oceans: a preliminary discussion.
The classical oceanographic problem of deducing the unknown constant in the dynamic method-the problem of the "level of no motion"-may be treated as a geophysical inverse problem. The unknown "barotropic" velocity may be chosen to satisfy an arbitrary number of conservation laws, subject to perfect geostrophic balance and with explicit use made of the relative errors in the observations. The solution obtained is one of minimum energy. A western North Atlantic region is used to demonstrate the power of the method.


Chronic benzodiazepine agonist treatment produces functional uncoupling of the gamma-aminobutyric acid-benzodiazepine receptor ionophore complex in cortical neurons.
We have investigated the effect of chronic flurazepam HCl treatment on the gamma-aminobutyric acid (GABA)A receptor complex in cultured mammalian cortical neurons. Chronic flurazepam (1-5 microM, for 1-10 days) treatment did not produce any changes in the morphological appearance or the cell protein content of cortical neurons. The basal binding of [3H]flunitrazepam, [3H] Ro15-1788, and [3H]Ro15-4513 was also not altered after the chronic treatment. However, chronic flurazepam treatment produced uncoupling between GABA and pentobarbital sites and the [3H]flunitrazepam binding site. The EC50 values of GABA and pentobarbital were not significantly altered after chronic flurazepam treatment; however, their Emax values were decreased by approximately 50%. The effect of chronic flurazepam treatment on the observed uncoupling was both time and concentration dependent. Furthermore, the binding of [3H]GABA and t-butylbicyclophosphoro[35S]thionate was also not altered by chronic flurazepam treatment. The effect of GABA on 36Cl influx was not altered after chronic flurazepam treatment; however, treatment significantly attenuated the ability of diazepam to enhance GABA-induced 36Cl influx. Chronic flurazepam-induced uncoupling and decreased diazepam efficacy were reversed by the concomitant presence of the benzodiazepine antagonist Ro15-1788, suggesting that these events are mediated via the benzodiazepine receptor site. Taken together, these results suggest that chronic benzodiazepine treatment produces uncoupling of GABA and pentobarbital sites from the benzodiazepine site and decreased coupling between the benzodiazepine site and GABA receptor-gated Cl- channels. The uncoupling and decreased efficacy may be due to an alteration in the levels of various alpha subunits and may be responsible for the tolerance associated with chronic benzodiazepine agonist treatment.


Pulsed high-intensity focused ultrasound enhances systemic administration of naked DNA in squamous cell carcinoma model: initial experience.
To determine whether exposures to pulsed high-intensity focused ultrasound can enhance local delivery and expression of a reporter gene, administered with systemic injection of naked DNA, in tumors in mice. The study was performed according to an approved animal protocol and in compliance with guidelines of the institutional animal care and use committee. Squamous cell carcinoma (SCC7) tumors were induced subcutaneously in both flanks of female C3H mice (n = 3) and allowed to grow to average size of 0.4 cm(3). In each mouse, one tumor was exposed to pulsed high-intensity focused ultrasound while a second tumor served as a control. Immediately after ultrasound exposure, a solution containing a cytomegalovirus-green fluorescent protein (GFP) reporter gene construct was injected intravenously via the tail vein. The mouse was sacrificed 24 hours later. Tissue specimens were viewed with fluorescence microscopy to determine the presence of GFP expression, and Western blot analysis was performed, at which signal intensities of expressed GFP were quantitated. A paired Student t test was used to compare mean values in controls with those in treated tumors. Histologic analyses were performed with specific techniques (hematoxylin-eosin staining, terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling) to determine whether tumor cells had been damaged by ultrasound exposure. GFP expression was present in all sections of tumors that received ultrasound exposure but not in control tumors. Results of signal intensity measurement at Western blot analysis showed expressed GFP to be nine times greater in ultrasound-exposed tumors (160.2 +/- 24.5 [standard deviation]) than in controls (17.4 +/- 11.8) (P = .004, paired Student t test). Comparison of histologic sections from treated tumors with those from controls revealed no destructive effects from ultrasound exposure. Local exposure to pulsed high-intensity focused ultrasound in tumors can enhance the delivery and expression of systemically injected naked DNA.


Management of macular puckers associated with retinal angiomas.
Monocular visual loss in four patients was caused by epiretinal macular membranes associated with peripheral retinal capillary angiomas or angioma-like lesions. Three patients had solitary vascular lesions and one had two discrete vascular lesions. All patients were in good health, with no evidence of the von Hippel-Lindau syndrome. In three patients, spontaneous peeling of the macular membranes and visual improvement occurred 2 to 6 months after obliteration of the angioma-like lesions with cryotherapy (two patients), or cryotherapy combined with argon laser (one patient). The fourth patient underwent a vitrectomy 8 months after the macular pucker had failed to respond to treatment of the vascular lesion.


M2 tumor-associated macrophage promoted DNA methylation in lung cancer metastasis via intensifying EZH2.
This study aimed to explore the interaction between the tumor-associated macrophage (TAM) and enhancer of zeste homolog 2 (EZH2) in tumor microenvironment of lung cancer are obscure. M2 type of TAM was induced by interleukin-4 (IL-4) and interleukin-13 (IL-13) in RAW264.7 cells. Subsequently, the co-culture system of the M2 RAW264.7 treating LLC-1 cells were constructed to evaluate the cell proliferation, migration and invasion abilities. On top of that, the M2 RAW264.7 was injected into the LLC-1 cells-bearing mice. Tumor growth and the number of metastatic nodes were observed. Moreover, DNA methylation, EZH2 expression, target genes of EZH2 and the M2 type TAM-related markers were detected in vivo and in vitro. Further experiments of EZH2 function in lung cancer were carried out by the addition of EZH2 inhibitor (GSK126) and si-EZH2. M2 type of TAM was induced with IL-4 and IL-13 with increased expression of CD206, CD68, CD163 and Arg1. Following co-culture with M2 type TAM, the proliferative, invasive, migrative abilities, tumor growth and metastasis, and the DNA methylation, EZH2 level were strengthened whereas the target genes of EZH2, including p21, CDKN2A, CDKN2B were reduced in LLC-1 cells and LLC-1 cell-bearing mice. Of note, GSK126 and si-EZH2 offset the M2 type TAM's effects, and inhibited the LLC-1 cell metastasis, DNA methylation and tumor growth. M2 type TAM promoted DNA methylation in LLC-1 cells and LLC-1 cell-bearing mice, which is related to the intensified EZH2.


A comparison of 2 influenza vaccine schedules in 6- to 23-month-old children.
Trivalent inactivated influenza vaccine (TIV) is recommended for all children ages 6 to 23 months. Delivering 2 doses of TIV at least 4 weeks apart to young children receiving this vaccine for the first time is challenging. We compared the immunogenicity and reactogenicity of the standard 2-dose regimen of TIV administered in the fall with an early schedule of a single spring dose followed by a fall dose of the same vaccine in healthy toddlers 6 to 23 months of age. Children were recruited in the spring to be randomized into either the standard or early schedule. An additional group was also enrolled in the fall as part of a nonrandomized standard comparison group. The 2002-2003 licensed TIV was administered in the spring; the fall 2003-2004 vaccine contained the same 3 antigenic components. Reactogenicity was assessed by parental diaries and telephone surveillance. Blood was obtained after the second dose of TIV for all children. The primary outcome measure was antibody response to influenza A/H1N1, A/H3N2, and B after 2 doses of vaccine, as determined by hemagglutination-inhibition titers > or =1:32 and geometric mean titer (GMT). Two hundred nineteen children were randomized to receive either the standard or early TIV schedule; 40 additional children were enrolled in the fall in the nonrandomized standard group. Response rates in the combined standard versus early groups were similar overall: 78% (GMT: 48) vs 76% (GMT: 57) to H1N1, 89% (GMT: 115) vs 88% (GMT: 129) to H3N2, and 52% (GMT: 24) vs 60% (GMT: 28) to B. Reactogenicity after TIV in both groups of children was minimal and did not differ by dose, age, or time between doses. Reaction rates were higher in those receiving TIV and concomitant vaccines compared with those receiving TIV alone. Overall rates of fever >38 degrees C axillary and injection-site pain, redness, or swelling were 5.4%, 3.1%, 0.9%, and 1.1%, respectively. When the spring and fall influenza vaccines had the same 3 antigenic components, the early vaccine schedule resulted in similar immunogenicity and reactogenicity compared with the standard schedule. When the vaccine components do not change between years, initiating influenza vaccine in the spring at the time of routine office visits would facilitate full immunization of children against influenza earlier in the season.


Differential Effects of Aripiprazole and Amisulpride on Negative and Cognitive Symptoms in Patients With First-Episode Psychoses.
Aripiprazole is hypothesized to have an effect on negative and cognitive symptoms in schizophrenia. Likewise, amisulpride is one of the only second-generation antipsychotics with which an effect on negative symptoms is reported. In the present study, we compare the effect of aripiprazole and amisulpride in initially antipsychotic-naïve patients with first-episode psychoses. Psychopathology and cognitive measures from two consecutive cohorts of antipsychotic-naïve first episode psychotic patients were obtained before and after 6 weeks of antipsychotic monotherapy with either aripiprazole or amisulpride. Matched healthy controls were included to account for retest effects on the cognitive measures. Analyses of variance (repeated-measures ANOVA) were performed to detect effect of time and possible cohort*time interactions. Longitudinal data was obtained from 47 and 48 patients treated for 6 weeks with amisulpride or aripiprazole, respectively. For the Wallwork negative symptom dimension, there was a cohort*time interaction [F (1, 93) = 4.29, p = 0.041] and a significant effect of time [F (1, 93) = 6.03, p = 0.016], which was driven by an improvement in patients treated with aripiprazole [t (47) = 4.1, p < 0.001] and not observed in patients treated with amisulpride (p > 0.5). For the eight cognitive measures, no cohort*time interaction was found and neither was cognitive improvement in any of the cohorts when accounting for retest effect. Patients treated with aripiprazole improved on negative symptoms, which was not the case for patients treated with amisulpride. This may point to a general effect of a partial D2 receptor agonist on negative symptoms in patients with first-episode psychoses. There was, however, no improvement in cognitive functions.


Finding the face in the crowd: an anger superiority effect.
Facial gestures have been given an increasingly critical role in models of emotion. The biological significance of interindividual transmission of emotional signals is a pivotal assumption for placing the face in a central position in these models. This assumption invited a logical corollary, examined in this article: Face-processing should be highly efficient. Three experiments documented an asymmetry in the processing of emotionally discrepant faces embedded in crowds. The results suggested that threatening faces pop out of crowds, perhaps as a result of a preattentive, parallel search for signals of direct threat.


Systems modeling and simulation applications for critical care medicine.
Critical care delivery is a complex, expensive, error prone, medical specialty and remains the focal point of major improvement efforts in healthcare delivery. Various modeling and simulation techniques offer unique opportunities to better understand the interactions between clinical physiology and care delivery. The novel insights gained from the systems perspective can then be used to develop and test new treatment strategies and make critical care delivery more efficient and effective. However, modeling and simulation applications in critical care remain underutilized. This article provides an overview of major computer-based simulation techniques as applied to critical care medicine. We provide three application examples of different simulation techniques, including a) pathophysiological model of acute lung injury, b) process modeling of critical care delivery, and c) an agent-based model to study interaction between pathophysiology and healthcare delivery. Finally, we identify certain challenges to, and opportunities for, future research in the area.


Genomic estimates of aneuploid content in glioblastoma multiforme and improved classification.
Accurate classification of glioblastoma multiforme (GBM) is crucial for understanding its biologic diversity and informing diagnosis and treatment. The Cancer Genome Atlas (TCGA) project identified four GBM classes using gene expression data and separately identified three classes using methylation data. We sought to integrate multiple data types in GBM classification, understand biologic features of the newly defined subtypes, and reconcile with prior studies. We used allele-specific copy number data to estimate the aneuploid content of each tumor and incorporated this measure of intratumor heterogeneity in class discovery. We estimated the potential cell of origin of individual subtypes and the euploid and aneuploid fractions using reference datasets of known neuronal cell types. There exists an unexpected correlation between aneuploid content and the observed among-tumor diversity of expression patterns. Joint use of DNA and mRNA data in ab initio class discovery revealed a distinct group that resembles the Proneural subtype described in a separate study and the glioma-CpG island methylator phenotype (G-CIMP+) class based on methylation data. Three additional subtypes, Classical, Proliferative, and Mesenchymal, were also identified and revised the assignment for many samples. The revision showed stronger differences in patient outcome and clearer cell type-specific signatures. Mesenchymal GBMs had higher euploid content, potentially contributed by microglia/macrophage infiltration. We clarified the confusion about the "Proneural" subtype that was defined differently in different prior studies. The ability to infer within-tumor heterogeneity improved class discovery, leading to new subtypes that are closer to the fundamental biology of GBM.


Characterization of the Zebrafish Glycine Receptor Family Reveals Insights Into Glycine Receptor Structure Function and Stoichiometry.
To study characterization of zebrafish glycine receptors (zGlyRs), we assessed expression and function of five α- and two ß-subunit encoding GlyR in zebrafish. Our qPCR analysis revealed variable expression during development, while in situ hybridizations uncovered expression in the hindbrain and spinal cord; a finding consistent with the reported expression of GlyR subunits in these tissues from other organisms. Electrophysiological recordings using Xenopus oocytes revealed that all five α subunits form homomeric receptors activated by glycine, and inhibited by strychnine and picrotoxin. In contrast, ß subunits only formed functional heteromeric receptors when co-expressed with α subunits. Curiously, the second transmembranes of both ß subunits were found to lack a phenylalanine at the sixth position that is commonly associated with conferring picrotoxin resistance to heteromeric receptors. Consistent with the absence of phenylalanines at the sixth position, heteromeric zGlyRs often lacked significant picrotoxin resistance. Subsequent efforts revealed that resistance to picrotoxin in both zebrafish and human heteromeric GlyRs involves known residues within transmembrane 2, as well as previously unknown residues within transmembrane 3. We also found that a dominant mutation in human GlyRα1 that gives rise to hyperekplexia, and recessive mutations in zebrafish GlyRßb that underlie the bandoneon family of motor mutants, result in reduced receptor function. Lastly, through the use of a concatenated construct we demonstrate that zebrafish heteromeric receptors assemble with a stoichiometry of 3α:2ß. Collectively, our findings have furthered our knowledge regarding the assembly of heteromeric receptors, and the molecular basis of ß subunit-conferred picrotoxin resistance. These results should aid in future investigations of glycinergic signaling in zebrafish and mammals.


The impact of volume overload on technique failure in incident peritoneal dialysis patients.
Technique failure in peritoneal dialysis (PD) can be due to patient- and procedure-related factors. With this analysis, we investigated the association of volume overload at the start and during the early phase of PD and technique failure. In this observational, international cohort study with longitudinal follow-up of incident PD patients, technique failure was defined as either transfer to haemodialysis or death, and transplantation was considered as a competing risk. We explored parameters at baseline or within the first 6 months and the association with technique failure between 6 and 18 months, using a competing risk model. Out of 1092 patients of the complete cohort, 719 met specific inclusion and exclusion criteria for this analysis. Being volume overloaded, either at baseline or Month 6, or at both time points, was associated with an increased risk of technique failure compared with the patient group that was euvolaemic at both time points. Undergoing treatment at a centre with a high proportion of PD patients was associated with a lower risk of technique failure. Volume overload at start of PD and/or at 6 months was associated with a higher risk of technique failure in the subsequent year. The risk was modified by centre characteristics, which varied among regions.


Exercise plasma boosts memory and dampens brain inflammation via clusterin.
Physical exercise is generally beneficial to all aspects of human and animal health, slowing cognitive ageing and neurodegeneration1. The cognitive benefits of physical exercise are tied to an increased plasticity and reduced inflammation within the hippocampus2-4, yet little is known about the factors and mechanisms that mediate these effects. Here we show that 'runner plasma', collected from voluntarily running mice and infused into sedentary mice, reduces baseline neuroinflammatory gene expression and experimentally induced brain inflammation. Plasma proteomic analysis revealed a concerted increase in complement cascade inhibitors including clusterin (CLU). Intravenously injected CLU binds to brain endothelial cells and reduces neuroinflammatory gene expression in a mouse model of acute brain inflammation and a mouse model of Alzheimer's disease. Patients with cognitive impairment who participated in structured exercise for 6 months had higher plasma levels of CLU. These findings demonstrate the existence of anti-inflammatory exercise factors that are transferrable, target the cerebrovasculature and benefit the brain, and are present in humans who engage in exercise.


Sleep loss impairs short and novel language tasks having a prefrontal focus.
Most cognitive tests administered during sleep loss are well rehearsed to remove practice effects. This can introduce tedium and a loss of novelty, which may be the key to the test's subsequent sensitivity to sleep loss, and why it may need only a few minutes administration before sleep loss effects are apparent. There is little evidence to show that any of these tests are actually affected by sleep loss is given de novo, without practice, but using a non-sleep deprived control group. Although the sleep deprivation literature advocates that short, novel and stimulating tests would not be expected to be sensitive to sleep loss, recent sleep loss findings using neuropsychological tests focussing on the prefrontal cortex, indicate that such tests may challenge this maxim. Twenty healthy young adults were randomly assigned to two groups: nil sleep deprivation (control). and 36h continuous sleep deprivation (SD). Two, novel, interesting and short (6 min) language tests, known (by brain imaging) to have predominantly a PFC focus, were given, once, towards the end of SD: (i) the Haylings test--which measures the capacity to inhibit strong associations in favour of novel responses, and (ii) a variant of the word fluency test--innovation in a verb-to-noun association. Subjects were exhorted to do their best. Compared with control subjects both tasks were significantly impaired by SD. As a check on the effects on the Haylings test, a repeat study was undertaken with 30 more subjects randomly divided as before. The outcome was similar. Linguistically, sleep loss appears to interfere with novel responses and the ability to suppress routine answers.


Cloning and characterization of the goadsporin biosynthetic gene cluster from Streptomyces sp. TP-A0584.
The biosynthetic gene cluster of goadsporin, a polypeptide antibiotic containing thiazole and oxazole rings, was cloned from Streptomyces sp. TP-A0584. The cluster contains a structural gene, godA, and nine god (goadsporin) genes involved in post-translational modification, immunity and transcriptional regulation. Although the gene organization is similar to typical bacteriocin biosynthetic gene clusters, each goadsporin biosynthetic gene shows low homology to these genes. Goadsporin biosynthesis is initiated by the translation of godA, and the subsequent cyclization, dehydration and acetylation are probably catalysed by godD, godE, godF, godG and godH gene products. godI shows high similarity to the 54 kDa subunit of the signal recognition particle and plays an important role in goadsporin immunity. Furthermore, four goadsporin analogues were produced by site-directed mutagenesis of godA, suggesting that this biosynthesis machinery is used for the heterocyclization of peptides.


Knowledge engineering for health: a new discipline required to bridge the "ICT gap" between research and healthcare.
Despite vast amount of money and research being channeled toward biomedical research, relatively little impact has been made on routine clinical practice. At the heart of this failure is the information and communication technology "chasm" that exists between research and healthcare. A new focus on "knowledge engineering for health" is needed to facilitate knowledge transmission across the research-healthcare gap. This discipline is required to engineer the bidirectional flow of data: processing research data and knowledge to identify clinically relevant advances and delivering these into healthcare use; conversely, making outcomes from the practice of medicine suitably available for use by the research community. This system will be able to self-optimize in that outcomes for patients treated by decisions that were based on the latest research knowledge will be fed back to the research world. A series of meetings, culminating in the "I-Health 2011" workshop, have brought together interdisciplinary experts to map the challenges and requirements for such a system. Here, we describe the main conclusions from these meetings. An "I4Health" interdisciplinary network of experts now exists to promote the key aims and objectives, namely "integrating and interpreting information for individualized healthcare," by developing the "knowledge engineering for health" domain.


Complex attentional control settings.
The visual system prioritizes information through a variety of mechanisms, including "attentional control settings" that specify features (e.g., colour) that are relevant to current goals. Recent work shows that these control settings may be more complex than previously thought, such that participants can monitor for independent features at different locations (Adamo, Pun, Pratt, & Ferber, 2008). However, this result leaves unclear whether these control settings affect early attentional selection or later target processing. We dissociated between these possibilities in two ways. In Experiment 1, participants were asked to determine whether a target object, which was preceded by an uninformative cue, matched one of two target templates (e.g., a blue vertical object or a green horizontal object). Participants monitored for independent features in the same location, but in different objects, which should reduce the effectiveness of the control setting if it is due to early attentional selection, but not if it is due to later target processing. In Experiment 2, we removed the ability of the cue to prime the target identity, which makes the opposite prediction. Together, the results suggest that complex attentional control settings primarily affect later target identity processing, and not early attentional selection.


Experience with fully 3D PET and implications for future high-resolution 3D tomographs.
The aim of this paper is to report on experience with 3D positron emission tomography (PET) in our institute where we have three 3D scanners, of which two operate exclusively in 3D mode (ECAT ART, EXACT 3D). Fully 3D PET requires attention to a number of factors which are not as problematic in 2D PET. Firstly, 3D tomographs designed for whole-body acquisition suffer from a large single-photon field of view, extending well beyond the coincidence field of view. Single photons from outside the coincidence field of view increase the dead time and random coincidence rates, and contribute scattered events. For brain studies, we have extended the lead side shielding at the ends of the tomograph to mitigate against these effects, and this has dramatically improved the count rate performance. This approach is not as effective for whole-body scanning. In addition, operating in 3D without septa necessitates new approaches to transmission scanning, as measurements using positron emitters such as 68Ge/68Ga have the unfavourable characteristics of high dead time and high scatter. Both of our fully 3D scanners use 137Cs for single-photon transmission measurements, although the data are treated differently. On the ECAT ART, a combination of physical and electronic collimation effectively reduces transmission scatter to acceptable levels. On the EXACT 3D physical collimation is not as readily implemented and therefore segmentation and reassignment of the histogrammed attenuation (mu) values is employed to produce unbiased attenuation correction factors in 3D. Many of the lessons learnt with these BGO (bismuth germanate) based tomographs will be applicable to the next generation of systems using faster detectors such as lutetium oxyorthosilicate (LSO).


Serum proteome profiling identifies novel and powerful markers of cystic fibrosis liver disease.
Cystic Fibrosis associated liver disease (CFLD) develops in approximately 30% of CF patients. However, routine sensitive diagnostic tools for CFLD are lacking. Within this study, we aimed to identify new experimental biomarkers for the detection of CFLD. 45 CF patients were included in the study and received transient elastography. Differential regulation of 220 different serum proteins was assessed in a subgroup of patients with and without CFLD. Most interesting candidate proteins were further quantified and validated by ELISA in the whole patient cohort. To assess a potential relation of biomarker expression to the degree of hepatic fibrosis, serum biomarkers were further determined in 18 HCV patients where liver histology was available. 43 serum proteins differed at least 2-fold in patients with CFLD compared to those without liver disease as identified in proteome profiling. In ELISA quantifications, TIMP-4 and Endoglin were significantly up-regulated in patients with CFLD as diagnosed by clinical guidelines or increased liver stiffness. Pentraxin-3 was significantly decreased in patients with CFLD. Serum TIMP-4 and Endoglin showed highest values in HCV patients with liver cirrhosis compared to those with fibrosis but without cirrhosis. At a cut-off value of 6.3 kPa, transient elastography compassed a very high diagnostic accuracy and specificity for the detection of CFLD. Among the biomarkers, TIMP-4 and Endoglin exhibited a high diagnostic accuracy for CFLD. Diagnostic sensitivities and negative predictive values were increased when elastography and TIMP-4 and Endoglin were combined for the detection of CFLD. Serum TIMP-4 and Endoglin are increased in CFLD and their expression correlates with hepatic staging. Determination of TIMP-4 and Endoglin together with transient elastography can increase the sensitivity for the non-invasive diagnosis of CFLD.


[Cellular regulation of human lymphocyte mitogenic factor release in vitro].
Production of the mitogenic facotr by human lymphocytes stimulated with phytohemagglutinin (PHA) was investigated. Anti-PHA antibodies were used for studying the culture medium mitogenic activity. The mitogenic factor production markedly increased after lymphocyte irradiation. When macrophages were eliminated, using iron powder, mitogenic factor generation was also increased. It was demonstrated that lymphocyte irradiation and macrophage elimination stimulated the mitogenic factor production throgh different mechanisms.