Characterization of the neuroanatomical distribution of agouti-related protein immunoreactivity in the rhesus monkey and the rat.
Agouti-related protein (AGRP) is a recently described homolog of the skin agouti protein. AGRP is transcribed primarily in the adrenal and hypothalamus and is a high affinity antagonist of the neural melanocortin-3 and melanocortin-4 receptors. The perikarya expressing AGRP messenger RNA are found in the arcuate nucleus of the rat and rhesus monkey. Using a polyclonal antibody against the pharmacologically active domain of AGRP (amino acids 83-132), we have also characterized the distribution of AGRP-immunoreactive neurons in both species. The major fiber tracts are conserved in both species, with dense projections originating in the arcuate nucleus and proceeding along the third ventricle. Dense fiber bundles are also visible in the paraventricular, dorsomedial, and posterior nuclei in the hypothalamus, in the bed nucleus of the stria terminalis, and in the lateral septal nucleus of the septal region. AGRP-containing neurons are not visualized in a number of areas, including portions of the amygdala, thalamus, and brain stem, that express MC3-R and MC4-R messenger RNA and receive innervation from POMC neurons that serve as the source of melanocortin agonists. Thus, AGRP is most likely to be involved in modulating a conserved subset of the physiological functions of central melanocortin peptides. Based on the particular distribution of AGRP neurons, those functions are likely to include the central control of energy homeostasis.


Time-of-flight-secondary ion mass spectrometry method development for high-sensitivity analysis of acid dyes in nylon fibers.
A minimally destructive technique for the determination of dyes in finished fibers provides an important tool for crime scene and other forensic investigations. The analytical power and the minimal sample consumption of time-of-flight-secondary ion mass spectrometric (TOF-SIMS) analysis provides the ability to obtain definitive molecular and elemental information relevant to fiber identification, including identification of dyes, from a very small volume of sample. For both fiber surface analysis and, with the aid of cryomicrotomy, fiber cross-section analysis, TOF-SIMS was used to identify various dyes in finished textile fibers. The analysis of C.I. Acid Blue 25 in nylon is presented as a representative example. The molecular ion of C.I. Acid Blue 25 with lower than 3% on weight-of-fiber (owf) dye loading cannot be identified on dyed nylon surfaces by TOF-SIMS using a Bi(3)(+) primary ion beam. Sputtering with C(60)(+) provided the ability to remove surface contamination as well as at least partially remove Bi-induced damage, resulting in a greatly improved signal-to-noise ratio for the Acid Blue 25 molecular ion. The use of C(60)(+) for damage removal in a cyclic manner along with Bi for data acquisition provided the ability to unambiguously identify Acid Blue 25 via its molecular ion at a concentration of 0.1% owf from both fiber surfaces and cross sections.


The cost-effectiveness of risedronate in the treatment of osteoporosis: an international perspective.
Risedronate, a bisphosphonate for treatment and prevention of osteoporosis, has been shown in several clinical trials to reduce the risk of fractures in postmenopausal women with osteoporosis. The cost-effectiveness of risedronate treatment has previously been evaluated within different country settings using different model and analysis approaches. The objective of this study was to assess the cost-effectiveness of risedronate in postmenopausal women in four European countries -- Sweden, Finland, Spain, and Belgium -- by making use of the same modelling framework and analysis setup. A previously developed Markov cohort model for the evaluation of osteoporosis treatments was used to estimate the cost-effectiveness of risedronate treatment. For each country, the model was populated with local mortality, fracture incidence, and cost data. Hip fractures, clinical vertebral fractures, and wrist fractures were included in the model. The incremental cost per quality-adjusted life years (QALY) gained from a 5-year intervention with risedronate compared to "no intervention" in 70-year-old women at the threshold of osteoporosis [T-score = -2.5 based on National Health and Nutrition Examination Survey (NHANES) III data] and previous vertebral fracture was estimated to be euro 860, euro 19,532, euro 11,782, and euro 32,515 in Sweden, Finland, Belgium, and Spain, respectively. Among 70-year-old women at the threshold of osteoporosis without previous fracture the estimated cost per QALY gained ranged from euro 21,148 (Sweden) to euro 80,100 (Spain). The differences in cost-effectiveness between countries are mainly explained by different costs (fracture and treatment costs), fracture risks, and discount rates. Based on cost per QALY gained threshold values found in the literature, the study results indicated risedronate to be cost effective in the treatment of elderly women with established osteoporosis in all the included countries. At a hypothetical threshold value of euro 40,000 per QALY gained, the results in this study indicate that risedronate is a cost-effective treatment in elderly women at the threshold of osteoporosis (i.e., a T-score of -2.5) with prevalent vertebral fractures in Sweden, Finland, Belgium, and Spain.


The role of post-mortem investigations in determining the cause of sudden unexpected death in infancy.
Several autopsy protocols have been suggested for investigating sudden unexpected deaths in infancy (SUDI). The aim of this study is to provide data on the utility of such post-mortem investigations from a large paediatric autopsy series to inform future policy. Retrospective analysis of >1500 consecutive post-mortem examinations carried out by specialist paediatric pathologists at a single centre during a 10-year period according to a common autopsy protocol that included the use of detailed ancillary investigations. SUDI was defined as the sudden unexpected death of an infant aged from 7 to 365 days. All data capture and cause of death classification were carried out according to defined criteria. Of 1516 paediatric post-mortem examinations, 546 presented as SUDI. In 202 infants (37%), death was explained by the autopsy findings. The other 344 cases (63%) remained unexplained. Of the explained deaths, over half (58%) were infective, most commonly due to pneumonia (22%). The component of the post-mortem examination that primarily determined the final cause of death was histological examination in 92 infants (46%), macroscopic examination in 61 (30%), microbiological investigations in 38 (19%) and clinical history in 10 (5%). This constitutes the largest single-institution autopsy study of SUDI. Ten years on from the Confidential Enquiry into Stillbirths and Deaths in Infancy (CESDI) SUDI studies, the ascertainment of a cause of death at autopsy has improved. However, with almost two thirds of SUDI remaining unexplained, alternative and/or additional diagnostic techniques are required to improve detection rates of identifiable causes of death at autopsy.


Do upper-body and centralized adiposity measure different aspects of regional body-fat distribution? Relationship to non-insulin-dependent diabetes mellitus, lipids, and lipoproteins.
Both central and upper-body adiposity are associated with high rates of type II non-insulin-dependent diabetes mellitus (NIDDM), high triglyceride levels, and low high-density lipoprotein (HDL) cholesterol levels. Previous data have also suggested that central and upper-body adiposity are relatively uncorrelated and hence may measure different aspects of regional body fat distribution. We assessed body mass index (BMI), the ratio of subscapular-to-triceps skinfold (STR), the ratio of waist-to-hip circumference (WHR), lipids, lipoproteins, and glucose tolerance in 738 Mexican Americans (ages 25-64 yr), who participated in the San Antonio Heart Study, a population-based study of diabetes and cardiovascular risk factors. NIDDM was diagnosed according to National Diabetes Data Group criteria. In general, STR and WHR were associated with high NIDDM rates, low HDL cholesterol levels, and high triglyceride levels, although WHR was somewhat more predictive of these than STR. In females, BMI, WHR, and STR all made independent contributions to prediction of NIDDM and HDL cholesterol; in males, WHR and STR both made independent contributions to prediction of triglyceride levels. This suggests that both indices may measure different aspects of body-fat distribution. Investigators should consider measuring both of these indicators of body-fat distribution in studies of diabetes and other cardiovascular risk factors, although if only a single measure is feasible, WHR appears to be preferable.


The effectiveness of intensive training for residents in interviewing. A randomized, controlled study.
Interviewing and the physician-patient relationship are crucial elements of medical care, but residencies provide little formal instruction in these areas. To determine the effects of a training program in interviewing on 1) residents' attitudes toward and skills in interviewing and 2) patients' physical and psychosocial well-being and satisfaction with care. Randomized, controlled study. Two university-based primary care residencies. 63 primary care residents in postgraduate year 1. A 1-month, full-time rotation in interviewing and related psychosocial topics. Residents and their patients were assessed before and after the 1-month rotation. Questionnaires were used to assess residents' commitment to interviewing and psychosocial medicine, estimate of the importance of such care, and confidence in their ability to provide such care. Knowledge of interviewing and psychosocial medicine was assessed with a multiple-choice test. Audiotaped interviews with real patients and videotaped interviews with simulated patients were rated for specific interviewing behaviors. Patients' anxiety, depression, and social dysfunction; role limitations; somatic symptom status; and levels of satisfaction with medical visits were assessed by questionnaires and telephone interviews. Trained residents were superior to untrained residents in knowledge (difference in adjusted post-test mean scores, 15.7% [95% CI, 11% to 20%]); attitudes, such as confidence in psychological sensitivity (difference, 0.61 points on a 7-point scale [CI, 0.32 to 0.91 points]); somatization management (difference, 0.99 points [CI, 0.64 to 1.35 points]); interviewing of real patients (difference, 1.39 points on an 11-point scale [CI, 0.32 to 2.45 points]); and interviewing (data gathering) of simulated patients (difference, 2.67 points [CI, 1.77 to 3.56 points]). Mean differences between the study groups were consistently in the appropriate direction for patient satisfaction and patient well-being, but effect sizes were too small to be considered meaningful. An intensive 1-month training rotation in interviewing improved residents' knowledge about, attitudes toward, and skills in interviewing.


Can Chunk Size Differences Explain Developmental Changes in Lexical Learning?
In three experiments, we investigated Hebb repetition learning (HRL) differences between children and adults, as a function of the type of item (lexical vs. sub-lexical) and the level of item-overlap between sequences. In a first experiment, it was shown that when non-repeating and repeating (Hebb) sequences of words were all permutations of the same words, HRL was slower than when the sequences shared no words. This item-overlap effect was observed in both children and adults. In a second experiment, we used syllable sequences and we observed reduced HRL due to item-overlap only in children. The findings are explained within a chunking account of the HRL effect on the basis of which we hypothesize that children, compared with adults, chunk syllable sequences in smaller units. By hypothesis, small chunks are more prone to interference from anagram representations included in the filler sequences, potentially explaining the item-overlap effect in children. This hypothesis was tested in a third experiment with adults where we experimentally manipulated the chunk size by embedding pauses in the syllable sequences. Interestingly, we showed that imposing a small chunk size caused adults to show the same behavioral effects as those observed in children. Departing from the analogy between verbal HRL and lexical development, the results are discussed in light of the less-is-more hypothesis of age-related differences in language acquisition.


Breathing control in neurological diseases.
Control of ventilation depends on a brainstem neuronal network that controls activity of the motor neurons innervating the respiratory muscles. This network includes the pontine respiratory group and the dorsal and ventral respiratory groups in the medulla, which contain neurons that fire primarily during inspiration, post-inspiration, or expiration. The ventral respiratory group includes the pre-Bötzinger complex, which contains neurokinin-1 receptor immunoreactive neurons critical for respiratory rhythmogenesis. Structural and degenerative disorders affecting this network produce abnormalities of respiration, including sleep apnea and various patterns of dysrhythmic breathing, not infrequently associated with disturbances of cardiovagal and sympathetic vasomotor control. This emphasizes the important interactions between the respiratory and cardiovascular control networks in the medulla. Common disorders associated with impaired cardiorespiratory control include brainstem stroke or compression, syringobulbia, Chiari malformation, high cervical spinal cord injuries, and multiple system atrophy. This review focuses on the functional organization of the respiratory control network and common causes of impaired control of respiration.


Process evaluation of the data-driven quality improvement in primary care (DQIP) trial: active and less active ingredients of a multi-component complex intervention to reduce high-risk primary care prescribing.
Two to 4% of emergency hospital admissions are caused by preventable adverse drug events. The estimated costs of such avoidable admissions in England were £530 million in 2015. The data-driven quality improvement in primary care (DQIP) intervention was designed to prompt review of patients vulnerable from currently prescribed non-steroidal anti-inflammatory drugs (NSAIDs) and anti-platelets and was found to be effective at reducing this prescribing. A process evaluation was conducted parallel to the trial, and this paper reports the analysis which aimed to explore response to the intervention delivered to clusters in relation to participants' perceptions about which intervention elements were active in changing their practice. Data generation was by in-depth interview with key staff exploring participant's perceptions of the intervention components. Analysis was iterative using the framework technique and drawing on normalisation process theory. All the primary components of the intervention were perceived as active, but at different stages of implementation: financial incentives primarily supported recruitment; education motivated the GPs to initiate implementation; the informatics tool facilitated sustained implementation. Participants perceived the primary components as interdependent. Intervention subcomponents also varied in whether and when they were active. For example, run charts providing feedback of change in prescribing over time were ignored in the informatics tool, but were motivating in some practices in the regular e-mailed newsletter. The high-risk NSAID and anti-platelet prescribing targeted was accepted as important by all interviewees, and this shared understanding was a key wider context underlying intervention effectiveness. This was a novel use of process evaluation data which examined whether and how the individual intervention components were effective from the perspective of the professionals delivering changed care to patients. These findings are important for reproducibility and roll-out of the intervention. ClinicalTrials.gov, NCT01425502 .


Development of a pilot project on data sharing among partners of the Italian Hub of Population Biobanks (HIBP): association between lipid profile and socio-demographic variables.
The Italian Hub of Population Biobanks (HIBP) includes both ongoing and completed studies that are heterogeneous in both their purpose and in the specimens collected. The heterogeneity in starting conditions makes sharing study data very difficult because of technical, ethical, and collection rights issues that hamper collaboration and synergy. With the aim of overcoming these difficulties and establishing the "proof-of-concept" that sharing studies is achievable among Italian collections, a data-sharing pilot project has been agreed to by HIBP members. Participants agreed to the general methodology and signed a shared Data Transfer Agreement. The biobanks involved were: EURAC (Micros study), CIG (GEHA project), CNESPS (FINE, MATISS, MONICA, OEC1998, ITR (Italian Twin Register), and IPREA studies, and MOLIBANK (Moli-Sani project). Biobank data were uploaded into a common database using a dedicated informatics infrastructure. Demographic data, and anthropometric and hematochemical parameters were shared for each record. Each biobank uploaded into the common database a dataset with a minimum of 1000 subjects, for a total of 5071 records. After a harmonization process, the final dataset included 3882 records. Subjects were grouped into three main geographic areas of Italy (North, Center, and South) and separate analyses were performed for men and women. The 3882 records were analyzed through multivariate logistic regression analysis. Results were expressed as odds ratios with 95% confidence interval. Results show several geographical differences in the lipidemic pattern, mostly regarding cholesterol-HDL, which represents a strong basis for further, deeper sample-based studies. This HIBP pilot study aimed to prove the feasibility of such collaborations and it provides a methodological prototype for future studies based on the participation in the partnership of well-established quality collections.


Sustained dysfunctional information processing in patients with Internet gaming disorder: 6-month follow-up ERP study.
Internet gaming disorder (IGD), defined as an inability to control Internet-based game play, leads to serious impairment in psychological and social functioning, but few studies have identified the neurophysiological characteristics of patients with IGD. The aim of this study was to determine neurophysiological markers of P300 components associated with changes in symptoms after outpatient management with pharmacotherapy in patients with IGD. The present prospective longitudinal study included 18 patients with IGD and 29 healthy controls. The patients with IGD completed a 6-month outpatient management program including selective serotonin reuptake inhibitor-based pharmacotherapy. Event-related potentials (ERPs) were acquired during the auditory oddball task. ERPs of the patients with IGD were recorded before and after treatment. Between-group differences and the pre-to-post treatment differences in P300 components were investigated using repeated-measures analysis of variance. The primary treatment outcome was a change in score on Young Internet Addiction Test between before and after treatment. At baseline assessments, the IGD group showed significantly reduced P300 amplitudes and delayed latencies at the midline centro-parietal site compared with those in the healthy controls. No significant changes in the P300 indices were observed between pre and post-treatment in the patients with IGD after 6 months of treatment, even though the patients with IGD exhibited significant improvements in their IGD symptoms. Furthermore, no significant difference in ERPs was observed between responders and nonresponders to a 6-month treatment in patients with IGD. These results suggest that reduced P300 amplitudes and delayed latencies are candidate endophenotypes in the pathophysiology of IGD.


Interoperable Readiness to Use Tourniquets by One's Familiarity With Different Models.
We investigated interoperability for a first aid provider to perform simulated use of three tourniquet models of maximal, moderate, and minimal familiarity. The experiment was focused on the tourniquets used by an expert who rendered aid on a manikin by using three models of tourniquet with different extents of familiarity: The familiarity with Combat Application Tourniquet (C-A-T) was maximal; that for Special Operations Forces Tactical Tourniquet (SOFTT) was moderate, and that for Military Emergency Tourniquet (MET) was minimal. Each model had a band-and-rod design. Interoperability changes as intermodel differences were beneficial or costly in that performances were improved or impaired in units of time, ease, blood, and pressure. Each model had 10 tests, and test order was randomized by model. The HapMed Leg Tourniquet Trainer simulated a limb amputation. In comparison of interoperability burdens, sums of 10 test durations by model for C-A-T, SOFTT, and MET were 38, 77, and 64 minutes, respectively; C-A-T was fastest (p ≤ .002, both). The sums of times to stop bleeding for C-A-T, SOFTT, and MET were 334, 953, and 826 seconds, respectively; C-A-T was fastest (p ≤ .0013, both). The sums of blood losses for C-A-T, SOFTT, and MET were 2105, 3287, and 4256mL, respectively; that for C-A-T was least (p ≤ .0005, both). The mean ease of use differed, with C-A-T being easiest (p ≤ .0046, both). The mean pressure differed, with C-A-T being higher than SOFTT (p = .0073). Timesaving strongly favored the model with which the user had maximal familiarity. In theory and simulation, interoperability bears costs in successfully attaining it, in maintaining it, and in failing either. The user's familiarity with tourniquet model was associated with improved interoperability as seen by improved performances. If multiple models are fielded, then organizations may plan on extra spending, supplying, training, and managing.


Growth factor-dependent phosphoinositide signalling.
A wide variety of messages, in the form of diffusible growth factors, hormones and cytokines, are carried throughout multicellular organisms to coordinate important physiological properties of target cells, such as proliferation, differentiation, migration, apoptosis and metabolism. Most messengers bind to cognate receptors on target cells, which initiate a characteristic cascade of reactions within the cell, ultimately leading to the desired response. The cellular response is defined by the combination of signalling components whose individual activity depends upon the number and type of surface receptors. Consequently the responses of different cell types to one or more stimuli can be quite disparate. A molecular understanding of the signalling pathways employed by each type of receptor therefore underlies the ability to rationalize many cellular functions and to correct disfunctions. As a well studied example of the primary signalling events that take place on the cytoplasmic leaflet of the plasma membrane following receptor activation, we will discuss how the widely expressed receptor for epidermal growth factor (EGF) causes the phosphorylation and hydrolysis of a signalling precursor, the membrane lipid phosphatidylinositol. This paradigm will be used to illustrate certain general principles of signalling, including formation of multienzyme complexes, compartmentation of second messengers and intermediates, and cross-talk between different signalling pathways.


Do shapes have feelings? Social attribution in children with autism spectrum disorder and attention-deficit/hyperactivity disorder.
Theory of mind (ToM) deficits are common in children with neurodevelopmental disorders (NDDs), such as autism spectrum disorder (ASD) and attention-deficit/hyperactivity disorder (ADHD), which contribute to their social and cognitive difficulties. The social attribution task (SAT) involves geometrical shapes moving in patterns that depict social interactions and is known to recruit brain regions from the classic ToM network. To better understand ToM in ASD and ADHD children, we examined the neural correlates using the SAT and functional magnetic resonance imaging (fMRI) in a cohort of 200 children: ASD (N = 76), ADHD (N = 74) and typically developing (TD; N = 50) (4-19 years). In the scanner, participants were presented with SAT videos corresponding to social help, social threat, and random conditions. Contrasting social vs. random, the ASD compared with TD children showed atypical activation in ToM brain areas-the middle temporal and anterior cingulate gyri. In the social help vs. social threat condition, atypical activation of the bilateral middle cingulate and right supramarginal and superior temporal gyri was shared across the NDD children, with between-diagnosis differences only being observed in the right fusiform. Data-driven subgrouping identified two distinct subgroups spanning all groups that differed in both their clinical characteristics and brain-behaviour relations with ToM ability.


Endovascular abdominal aortic aneurysm repair in 144 patients: correlation of aneurysm size, proximal aortic neck length, and procedure-related complications.
During endovascular abdominal aortic aneurysm (AAA) repair, larger aneurysms often present formidable anatomic challenges to the insertion of the delivery catheter and graft deployment. The authors sought to evaluate whether large-diameter aneurysms and those with short proximal aortic necks might be associated with a higher frequency of insertion-related and short-term complications. From October 1999 to August 2000, 144 patients underwent elective endovascular graft placement for infrarenal AAA disease at the authors' institution. These patients were treated with use of the AneuRx bifurcated endoprosthesis. AAA size (maximum aneurysm diameter) and proximal aortic neck length were compared to estimated blood loss, operative time, accuracy of graft placement, presence of endoleak, intraoperative and postoperative complications (such as limb occlusion or vascular injury), length of hospital stay, and mortality. Statistical methods included correlation analysis and logistic regression. There were 121 men and 23 women whose aneurysms ranged in size from 3 cm to 9.8 cm (mean, 5.6 cm; 95% CI, 5.4-5.8 cm). Endograft insertion was successful in all cases. There were three deaths within 30 days (2.1%) and seven deaths overall (4.9%). There were 43 intraoperative complications (29.9%) in 31 patients (21.5%), most of them minor. Patients with major intraoperative complications had significantly longer procedure times than those without complications (337 vs. 149 min; P <.0001). In the postoperative period (within 30 days), 31 complications (21.5%) occurred in 28 patients (19.4%), again most of them minor. AAA size was unrelated in any way to the rate of complications, but short proximal aortic neck length was associated with more serious intraoperative and postoperative complications (P =.0404 and P =.0230, respectively), and decreased 30-day and overall survival (P =.0240 and P =.0152, respectively). Endovascular repair of large AAAs can be challenging; however, the size of the AAA does not influence the rate of complications. A short proximal aortic neck is the only significant risk factor for more serious complications.


Microcolinearity and genome evolution in the AdhA region of diploid and polyploid cotton (Gossypium).
Genome sizes vary by several orders of magnitude, driven by mechanisms such as illegitimate recombination and transposable element proliferation. Prior analysis of the CesA region in two cotton genomes that diverged 5-10 million years ago (Ma), and acquired a twofold difference in genome size, revealed extensive local conservation of genic and intergenic regions, with no evidence of the global genome size difference. The present study extends the comparison to include BAC sequences surrounding the gene encoding alcohol dehydrogenase A (AdhA) from four cotton genomes: the two co-resident genomes (A(T) and D(T)) of the allotetraploid, Gossypium hirsutum, as well as the model diploid progenitors, Gossypium arboreum (A) and Gossypium raimondii (D). In contrast to earlier work, evolution in the AdhA region reflects, in a microcosm, the overall difference in genome size, with a nearly twofold difference in aligned sequence length. Most size differences may be attributed to differential accumulation of retroelements during divergence of the genome diploids from their common ancestor, but in addition there has been a biased accumulation of small deletions, such that those in the smaller D genome are on average twice as large as those in the larger A genome. The data also provide evidence for the global phenomenon of 'genomic downsizing' in polyploids shortly after formation. This in part reflects a higher frequency of small deletions post-polyploidization, and increased illegitimate recombination. In conjunction with previous work, the data here confirm the conclusion that genome size evolution reflects many forces that collectively operate heterogeneously among genomic regions.


Community-associated methicillin-resistant Staphylococcus aureus immune evasion and virulence.
Staphylococcus aureus is a significant cause of human infections globally. Methicillin-resistant S. aureus (MRSA) emerged in the early 1960s and is now endemic in most healthcare facilities. Although healthcare-associated MRSA infections remain a major problem in most industrialized countries, those caused by community-associated MRSA (CA-MRSA) are now the most abundant cause of bacterial infections in the community in some parts of the world, such as the United States. The basis for the emergence and subsequent success of CA-MRSA is incompletely defined. However, the ability of the pathogen to cause disease in otherwise healthy individuals is likely attributed, in part, to its ability to circumvent killing by the innate immune system, which includes survival after phagocytosis by neutrophils. In this review, we discuss the role of neutrophils in host defense against S. aureus and highlight progress made toward understanding mechanisms of CA-MRSA virulence and pathogenesis.


Multicellularity Drives the Evolution of Sexual Traits.
From the male peacock's tail plumage to the floral displays of flowering plants, traits related to sexual reproduction are often complex and exaggerated. Why has sexual reproduction become so complicated? Why have such exaggerated sexual traits evolved? Early work posited a connection between multicellularity and sexual traits such as anisogamy (i.e., the evolution of small sperm and large eggs). Anisogamy then drives the evolution of other forms of sexual dimorphism. Yet the relationship between multicellularity and the evolution of sexual traits has not been empirically tested. Given their extensive variation in both multicellular complexity and sexual systems, the volvocine green algae offer a tractable system for understanding the interrelationship of multicellular complexity and sex. Here we show that species with greater multicellular complexity have a significantly larger number of derived sexual traits, including anisogamy, internal fertilization, and secondary sexual dimorphism. Our results demonstrate that anisogamy repeatedly evolved from isogamous multicellular ancestors and that anisogamous species are larger and produce larger zygotes than isogamous species. In the volvocine algae, the evolution of multicellularity likely drives the evolution of anisogamy, and anisogamy subsequently drives secondary sexual dimorphism. Multicellularity may set the stage for the overall diversity of sexual complexity throughout the Tree of Life.


Incidence and outcome of BRCA mutations in unselected patients with triple receptor-negative breast cancer.
To investigate the incidence of germline and somatic BRCA1/2 mutations in unselected patients with triple-negative breast cancer (TNBC) and determine the prognostic significance of carrying a mutation. DNA was obtained from 77 TNBC and normal tissues. BRCA1/2 exons/flanking regions were sequenced from tumor and patients classified as mutant or wild type (WT). Sequencing was repeated from normal tissue to identify germline and somatic mutations. Patient characteristics were compared with chi-square. Survival was estimated by Kaplan-Meier method and compared with log-rank. Cox proportional hazards models were fit to determine the independent association of mutation status with outcome. Median age was 51 years (27-83 years). Fifteen patients (19.5%) had BRCA mutations: 12 (15.6%) in BRCA1 (one somatic), and 3 (3.9%) in BRCA2. Patients with BRCA mutations tended to be younger than WT, (P = 0.005). Grade, histology, and stage were not associated with mutation status. At a median follow-up of 43 months (7-214 months), there were 33 (42.9%) recurrences and 35 (45.5%) deaths. Five-year recurrence-free survival estimates were 51.7% for WT versus 86.2% for patients with mutations, (P = 0.031); and 5-year overall survival estimates were 52.8% for WT versus 73.3% for patients with mutations (P = 0.225). After adjustment, patients with BRCA mutations had a significantly better RFS (HR: 0.19, 95% CI: 0.045-0.79, P = 0.016) compared with WT. In this unselected cohort of TNBC, we found a 19.5% incidence of BRCA mutations. Genetic testing should be discussed with patients with TNBC. Patients with TNBC with BRCA mutations had a significantly lower risk of relapse.


N1-Benzoyl-N2-[1-(1-naphthyl)ethyl]-trans-1,2-diaminocyclohexanes: Development of 4-chlorophenylcarboxamide (calhex 231) as a new calcium sensing receptor ligand demonstrating potent calcilytic activity.
A structure-activity relationship (SAR) study was performed principally at the N1 position of N1-arylsulfonyl-N2-[1-(1-naphthyl)ethyl]-trans-1,2-diaminocyclohexanes, a new family of calcilytics acting at the calcium sensing receptor (CaSR). The most active compound in this series was the 4-(trifluoromethoxy)benzenesulfonyl derivative 7e, which displayed an IC50 of 5.4 +/- 0.5 microM with respect to the inhibition of calcium-induced tritiated inositol phosphate ([3H]IP) accumulation in Chinese hamster ovarian (CHO) cells expressing the CaSR. Replacement of the sulfonamide linkage of this compound by a carboxamide led to a 6-fold increase in activity (7m, IC50 = 0.9 +/- 0.2 microM). Among the carboxamides synthesized, one of the most active compounds was the 4-chlorophenylcarboxamide (1S,2S,1'R)-7n (Calhex 231, IC50 = 0.33 +/- 0.02 microM). The absolute configuration of (1S,2S,1'R)-7n was deduced from an X-ray crystallographic study of one of the diastereomers of compound 7d. The stereochemical preference for the (1S,2S,1'R)-isomers can be rationalized on the basis of a three-dimensional model of the calcilytic binding pocket of the CaSR. Removal of the C-1' methyl group or replacement of the 1-naphthyl group by a 2-naphthyl or biphenyl moiety led to appreciable loss of calcilytic activity. Compounds 7e, 7m, and Calhex 231 did not stimulate [3H]IP accumulation in CHO cells expressing or not expressing the CaSR.


Annexin A1 Deficiency does not Affect Myofiber Repair but Delays Regeneration of Injured Muscles.
Repair and regeneration of the injured skeletal myofiber involves fusion of intracellular vesicles with sarcolemma and fusion of the muscle progenitor cells respectively. In vitro experiments have identified involvement of Annexin A1 (Anx A1) in both these fusion processes. To determine if Anx A1 contributes to these processes during muscle repair in vivo, we have assessed muscle growth and repair in Anx A1-deficient mouse (AnxA1-/-). We found that the lack of Anx A1 does not affect the muscle size and repair of myofibers following focal sarcolemmal injury and lengthening contraction injury. However, the lack of Anx A1 delayed muscle regeneration after notexin-induced injury. This delay in muscle regeneration was not caused by a slowdown in proliferation and differentiation of satellite cells. Instead, lack of Anx A1 lowered the proportion of differentiating myoblasts that managed to fuse with the injured myofibers by days 5 and 7 after notexin injury as compared to the wild type (w.t.) mice. Despite this early slowdown in fusion of Anx A1-/- myoblasts, regeneration caught up at later times post injury. These results establish in vivo role of Anx A1 in cell fusion required for myofiber regeneration and not in intracellular vesicle fusion needed for repair of myofiber sarcolemma.


Vascular endothelial growth factor-mediated angiogenesis inhibition and postoperative wound healing in rats.
Vascular endothelial growth factor (VEGF) is an angiogenic factor that acts by binding to specific high-affinity tyrosine kinase receptors. SU5416 is an antiangiogenic agent that acts as a potent and selective inhibitor of the VEGF Flk-1/KDR receptor tyrosine kinase. SU5416 has been shown to inhibit VEGF-dependent mitogenesis of human endothelial cells and to decrease the growth of xenografts of melanoma, lung carcinoma, ovarian carcinoma, and gliomas. The effect of pre- or perioperative use of this drug on angiogenesis and wound healing in the postoperative setting has not been shown. We sought to analyze the efficacy and safety with respect to functional dosing of SU5416 in the setting of wound healing. This represents an important step forward in the use of this and similar drugs in the perioperative setting of treatment for multiple types of cancers. The use of an inhibitor of VEGF receptors such as SU5416 is distinct and it is likely complementary to other agents in the treatment of such cancers. We injected 8-week-old male Sprague-Dawley rats with SU5416 (8 or 12 mg/kg) or dimethyl sulfoxide intraperitoneally, daily for 14 days. We then performed a right pulmonary lobectomy and 6-mm full-thickness punch biopsies of the back. Tissue perfusion measured via laser Doppler on Postoperative Day 2 was 1.65, 1.22, and 1.14 perfusion units (P < 0.0004) for control, 8 mg/kg, and 12 mg/kg groups, respectively. We successfully treated a murine model with functional doses of the anti-VEGF drug SU5416 so as to achieve decreased vascularity and blood flow in postoperative wounds. There was no effect on gross wound healing or infection in either control or treatment groups. Also, no drug-related impairment of histologic healing or decrease in wound tensile strength was demonstrated at either 6 or 14 days. Preoperative therapy with functional dosing of SU5416 does not appear to have any major effect on postoperative morbidity or mortality in rats. We additionally conclude that preoperative therapy with SU5416 should be investigated further with careful attention to wound integrity.


Femoral subchondral bone properties of patients with cam-type femoroacetabular impingement.
Morphological deformities of the hip, such as femoroacetabular impingement (FAI) may be responsible for up to 80% of hip osteoarthritis. In cam type FAI, the pathomechanism has been attributed to repeated abnormal contact between the femur and the antero-superior acetabular rim, resulting in cartilage and labrum degeneration. Subchondral bone stiffness likely plays a major role in the process, but little is known of the mechanical properties of the cam deformity. The purpose of this study was to determine tissue modulus and the trabecular micro-architecture of the subchondral bone of the cam deformity of patients undergoing resection surgery as well as comparing these parameters to healthy aged matched controls. Twelve osteochondral bone biopsies were obtained from symptomatic FAI patients and ten osteochondral control specimens were harvested from cadaveric femurs. A combination of mechanical testing, micro-CT and finite element (FE) analysis were used to determine tissue modulus, bone volume fraction, trabecular thickness, trabecular and spacing, and trabecular number. The mean tissue modulus of the cam-type FAI deformities (E = 5.4 GPa) was significantly higher than normal controls (E = 2.75 GPa, P = 0.038), but no statistically significant differences were found in bone micro-architectural parameters. The data suggests that subchondral bone of the cam deformity consists of older secondary mineralized bone. This supports the notion that the cam deformity is a primary malformation with intrinsic biomechanical abnormalities rather than a secondary deformity as part of the degenerative process of the covering cartilage or remodeling due to repeated impingement.


New insights into changes in corneal thickness in healthy mountaineers during a very-high-altitude climb to Mount Muztagh Ata.
To investigate the effect of very high altitude and different ascent profiles on central corneal thickness (CCT). Twenty-eight healthy mountaineers were randomly assigned to 2 different ascent profiles during a medical research expedition to Mount Muztagh Ata (7546 m) in western China. Group 1 was allotted a shorter acclimatization time prior to ascent to 6265 m. The main outcome measure was CCT. Secondary outcome measures were oxygen saturation (SpO(2)) and symptom assessments of acute mountain sickness (cerebral acute mountain sickness score). Examinations were performed at 490, 4497, 5533, and 6265 m. Central corneal thickness increased in both groups with increasing altitude and decreased after descent. In group 1 (with the shorter acclimatization), mean CCT increased from 537 to 572 microm. Mean CCT in group 2 increased from 534 to 563 microm (P = .048). The amount of decrease in SpO(2) paralleled the increase in CCT. There was no significant decrease in visual acuity. There was a significant correlation between CCT and cerebral acute mountain sickness score when controlled for SpO(2) and age. Corneal swelling during high-altitude climbs is promoted by low SpO(2). Systemic delivery of oxygen to the anterior chamber seems to play a greater role in corneal oxygenation than previously thought. Adhering to a slower ascent profile results in less corneal edema. Visual acuity in healthy corneas is not adversely affected by edema at altitudes of up to 6300 m. Individuals with more acute mountain sickness-related symptoms had thicker corneas, possibly due to their higher overall susceptibility to hypoxia.


Efficient cryopreservation of testicular tissue: effect of age, sample state, and concentration of cryoprotectant.
To study the effect of age and sample state on cryopreservation of testicular tissue, evaluate toxicity of commonly used cryoprotectants (CPs), and determine their optimal concentration for use. Prospective experimental study. Academic research unit. Patients with prostate carcinoma undergoing orchidectomy. We also studied immature and adult male Holtzman rats. Toxicity of CPs before freezing, morphology, and relative viability after freezing were evaluated for rat testicular cell suspensions (CS) and tubular fragments (TUB). Relative viability of adult human testicular CS and TUB after thaw was evaluated. Human TUB were cultured after thaw for 48 hours in medium containing epidermal growth factor (EGF), and effects on viability, morphology, and gene expression were determined. Viability and ploidy were measured with flow cytometry, postthaw cryodamage of immature rat tissue was studied by transmission electron microscopy, cell proliferation and differentiation were evaluated by immunohistochemistry and by real-time polymerase chain reaction. Immature testicular tissue was more susceptible to toxic assault by CP than adult tissue and displayed cell-specific sensitivity to CP, with glycerol, dimethyl sulfoxide and ethylene glycol being effective in protecting spermatid (1N), spermatogonia (2N) and spermatocyte (4N) populations respectively. Preservation as TUB may be preferred over CS and DMSO is an effective CP for immature and ethylene glycol for adult testicular tissue. Differential sensitivity of immature testicular tissue to CPs warrants judicious selection of CP on the basis of end application for prepubertal tissue.


Development of search strategies for systematic reviews: validation showed the noninferiority of the objective approach.
Different approaches can be adopted for the development of search strategies of systematic reviews. The objective approach draws on already established text analysis methods for developing search filters. Our aim was to determine whether the objective approach for the development of search strategies was noninferior to the conceptual approach commonly used in Cochrane reviews (CRs). We conducted a search for CRs published in the Cochrane Library. The studies included in the CRs were searched for in MEDLINE and represented the total set. We then tested whether references previously removed could be identified via the objective approach. We also reconstructed the original search strategies from the CRs to determine why references could not be identified by the objective approach. As we performed the validation of the search strategies without study filters, we used only sensitivity as a quality measure and did not calculate precision. The objective approach yielded a mean sensitivity of 96% based on 13 searches. The noninferiority test showed that this approach was noninferior to the conceptual approach used in the CRs (P < 0.002). An additional descriptive analysis showed that the original MEDLINE strategies could identify only 86% of all references; however, this lower sensitivity was largely due to one CR. To the best of our knowledge, our findings indicate for the first time that the objective approach for the development of search strategies is noninferior to the conceptual approach.


Interferon regulatory factor 7 mediates activation of Tap-2 by Epstein-Barr virus latent membrane protein 1.
Transporter associated with antigen processing 2 (Tap-2) is responsible for ATP-dependent transport of peptides from the cytosol to the endoplasmic reticulum, where peptides bind to newly synthesized human leukocyte antigen (HLA) class I molecules, which are essential for cellular immune responses. Epstein-Barr virus (EBV) latent membrane protein 1 (LMP-1) has been shown to induce the expression of Tap-2. In this study, the induction of endogenous Tap-2 by LMP-1 is shown to be associated with and requires the expression of interferon regulatory factor 7 (IRF-7). In DG75 Burkitt's lymphoma (BL) cells, in which LMP-1 induces the expression of IRF-7, LMP-1 induced endogenous Tap-2, and ectopic expression of IRF-7 could enhance the induction. In Akata BL cells, in which LMP-1 could not induce IRF-7, LMP-1 could not induce Tap-2. Addition of IRF-7, which complements the defect in Akata cells, could stimulate the expression of Tap-2. Furthermore, LMP-1 and IRF-7A but not other IRF-7 splicing variants could activate endogenous Tap-2. A Tap-2 promoter reporter construct could be activated by the overexpression of IRF-7A. The activation could be specifically enhanced by LMP-1 and was dependent on an intact interferon-stimulated response element (ISRE) present in the Tap-2 promoter. Also, IRF-7 can bind to the Tap-2 promoter under physiological conditions in vivo, as shown by formaldehyde cross-linking, as well as to the Tap-2 ISRE in vitro, as shown by gel mobility shift assays. Furthermore, LMP-1 facilitates the phosphorylation and nuclear translocation of IRF-7. These data point to the role of IRF-7 as a secondary mediator of LMP-1-activated signal transduction for Tap-2 as follows: LMP-1 stimulates the expression of IRF-7 and facilitates its phosphorylation and nuclear translocation, and then the activated IRF-7 mediates the activation of the cellular Tap-2 gene. The induction of Tap-2 by IRF-7 and LMP-1 may have an important implication for the immune response to EBV and its persistence in vivo.


Response Evaluation Criteria in Cancer of the Liver (RECICL) proposed by the Liver Cancer Study Group of Japan (2009 Revised Version).
The World Health Organization (WHO) criteria and Response Evaluation Criteria in Solid Tumors (RECIST) are inappropriate to assess the direct effects of treatment on the hepatocellular carcinoma (HCC) by locoreginal therapies such as radiofrequency ablation (RFA) and transcatheter arterial chemoembolization (TACE). Therefore, establishment of response evaluation criteria solely devoted for HCC is needed urgently in the clinical practice as well as in the clinical trials of HCC treatment, such as molecular targeted therapies, which cause necrosis of the tumor. Response Evaluation Criteria in Cancer of the Liver (RECICL) was revised in 2009 by Liver Cancer Study Group of Japan based on the 2004 version of RECICL, which was commonly used in Japan. Major revised points of the RECICL 2009 is to provide TE4a (Complete response with enough ablative margin) and TE4b (complete response without enough ablative margin) for local ablation therapy. Second revised point is that setting the timing at which the overall treatment effects are assessed. Third point is that emergence of new lesion in the liver is regarded as progressive disease, different from 2004 version. Finally, 3 tumor markers including alpha-fetoprotein (AFP) and AFP-L3 and des-gamma-carboxy protein (DCP) were also added for the overall treatment response. We hope this new treatment response criteria, RECICL, proposed by Liver Cancer Study Group of Japan will benefit the HCC treatment response evaluation in the setting of the daily clinical practice and clinical trials as well not only in Japan, but also internationally.


Identification of prohormones and pituitary neuropeptides in the African cichlid, Astatotilapia burtoni.
Cichlid fishes have evolved remarkably diverse reproductive, social, and feeding behaviors. Cell-to-cell signaling molecules, notably neuropeptides and peptide hormones, are known to regulate these behaviors across vertebrates. This class of signaling molecules derives from prohormone genes that have undergone multiple duplications and losses in fishes. Whether and how subfunctionalization, neofunctionalization, or losses of neuropeptides and peptide hormones have contributed to fish behavioral diversity is largely unknown. Information on fish prohormones has been limited and is complicated by the whole genome duplication of the teleost ancestor. We combined bioinformatics, mass spectrometry-enabled peptidomics, and molecular techniques to identify the suite of neuropeptide prohormones and pituitary peptide products in Astatotilapia burtoni, a well-studied member of the diverse African cichlid clade. Utilizing the A. burtoni genome, we identified 148 prohormone genes, with 21 identified as a single copy and 39 with at least 2 duplicated copies. Retention of prohormone duplicates was therefore 41 %, which is markedly above previous reports for the genome-wide average in teleosts. Beyond the expected whole genome duplication, differences between cichlids and mammals can be attributed to gene loss in tetrapods and additional duplication after divergence. Mass spectrometric analysis of the pituitary identified 620 unique peptide sequences that were matched to 120 unique proteins. Finally, we used in situ hybridization to localize the expression of galanin, a prohormone with exceptional sequence divergence in cichlids, as well as the expression of a proopiomelanocortin, prohormone that has undergone an additional duplication in some bony fish lineages. We characterized the A. burtoni prohormone complement. Two thirds of prohormone families contain duplications either from the teleost whole genome duplication or a more recent duplication. Our bioinformatic and mass spectrometric findings provide information on a major vertebrate clade that will further our understanding of the functional ramifications of these prohormone losses, duplications, and sequence changes across vertebrate evolution. In the context of the cichlid radiation, these findings will also facilitate the exploration of neuropeptide and peptide hormone function in behavioral diversity both within A. burtoni and across cichlid and other fish species.


Chemotherapy Education: An Interprofessional Approach to Standardizing Processes and Improving Nurse and Patient Satisfaction.
A lack of standardization in chemotherapy patient education practices can lead to decreased efficiency and satisfaction for nurse educators and uncertainty and anxiety for patients. The goal was to determine whether standardizing chemotherapy education practices improved nurse and patient satisfaction. An interprofessional team was formed to standardize the chemotherapy education process and reduce variation in teaching. Anonymous, self-administered questionnaires assessed satisfaction in chemotherapy education among nurses and patients pre- and postimplementation. Significant improvement in nursing staff satisfaction postimplementation was observed across all individual construct measures, with the average overall score increasing from 3.4 to 4.3. Patient satisfaction scores were high in the pre- and postimplementation phases (average overall score of 4.3 and 4.1, respectively).


Warfarin pharmacogenetics: polymorphisms of the CYP2C9, CYP4F2, and VKORC1 loci in a genetically admixed Omani population.
This is the first study to evaluate the spectrum and prevalence of dose-predictive genetic polymorphisms of the CYP2C9, CYP4F2 and VKORC1 loci together, in a geographically defined, ethnically admixed healthy adult Omani population sharing common lifestyle/environmental factors. Since the present-day Omani population is the result of an admixture of Caucasian, African and Asian ancestries, we compared the pharmacogenetic profile of these three loci in this population. Interestingly, the Omani pharmacogenetic profile, in terms of allele and genotype distribution, has values that are intermediate between Caucasians and African Americans, the African admixture further substantiated by the presence of the CYP2C9*8 allele. However, limitations and usefulness of such comparisons warrant caution, as the data from pharmacogenetic literature do not always represent bona fide population categories. Furthermore, definition of study population based on microgeographical scale would be more appropriate in pharmacogenetic research rather than the flawed racial, ethnic, or social categorizations since pharmacogenetic variation is clinal, and genetic influences will be further altered by lifestyle and environmental factors.


Total parenteral nutrition-associated alterations in hepatobiliary function and histology in rats: is light exposure a clue?
Light exposure of amino acids, especially in the presence of photosensitizers such as riboflavin, has been shown to result in photoproducts that are toxic both in vivo and in vitro. Provision of photooxidized single amino acids has been shown to result in hepatic dysfunction in both gerbils and rats. However, studies of the effects of light exposure of complete nutrient mixtures (glucose-amino acid-vitamins) on hepatobiliary responses are limited. These are important because, in clinical practice, total parenteral nutrition (TPN) solutions are continually exposed to light and because hepatic dysfunction is the most common metabolic aberration associated with TPN. Studies were conducted to compare the effects of TPN that had been exposed to light (+L) or protected from light (-L) on hepatobiliary function of rats. TPN solutions were either exposed to light or protected from light for 24 h and then infused into rats for 10 d. Data for enterally fed rats are also shown for comparison with parenterally fed animals. +L animals lost more weight and had lower bile flow, higher taurocholate output in bile, higher biliary osmolarity, and higher inorganic phosphate in bile. Hepatic histology demonstrated scattered foci of necrosis in eight of the eight +L animals and in only one of eight -L animals. These data demonstrate that protection of TPN solutions from light minimizes TPN-associated alterations in hepatobiliary function and histology. These histologic changes observed in the +L animals are in contrast to previous reports of TPN-induced histologic changes, suggesting a different mechanism. Our observations suggest that consideration be given to the implications of exposing TPN solutions to light.


Regulation of the angiopoietin-Tie ligand-receptor system with a novel splice variant of Tie1 reduces the severity of murine arthritis.
To determine the function of the angiopoietin (Ang)-Tie ligand-receptor system, and to assess the effect of Tie1-751, a naturally occurring extracellular domain of the Tie1 receptor derived by alternative splicing, in an in vivo model of RA. In the murine CIA model, expression of endogenous Ang1, Ang2, Tie1 and Tie2 in whole paws was analysed by quantitative RT-PCR. To assess the effect of inhibition of the Ang-Tie axis, Tie1-751 was expressed and fused to the Fc fragment of human IgG1. The effect of Tie1-751-Fc on human umbilical vein endothelial cell (HUVEC) cytoprotection and migration in response to Ang1, either alone or in combination with VEGF, was investigated. Furthermore, an in vitro angiogenesis assay was used to determine the effect of Tie1-751-Fc on Ang1-mediated angiogenesis. Finally, Tie1-751-Fc was administered in CIA, and the effects on clinical disease and joint architecture of hind foot specimens were determined. Gene expression levels of Ang1, Ang2, and receptors Tie1 and Tie2 in whole paws were significantly increased during the progression of arthritis. Tie1-751-Fc significantly inhibited HUVEC cytoprotection and migration in response to Ang1 alone, or Ang1 in combination with VEGF. Tie1-751-Fc also significantly inhibited angiogenesis induced by a combination of Ang1 plus VEGF. Finally, Tie1-751-Fc, when administered intra-peritoneally to arthritic mice, reduced clinical signs of arthritis, damage to joint architecture and infiltration of blood vessels into the synovium. Our data demonstrate that the Ang-Tie ligand-receptor system is dysregulated in CIA. Tie1-751, a novel splice variant of the Tie1 receptor, inhibits Ang1/VEGF signalling, suggesting that Ang inhibition may be of therapeutic benefit in inflammatory arthritis.


Multivariate Pattern Analysis of Volumetric Neuroimaging Data and Its Relationship With Cognitive Function in Treated HIV Disease.
Accurate prediction of longitudinal changes in cognitive function would potentially allow for targeted intervention in those at greatest risk of cognitive decline. We sought to build a multivariate model using volumetric neuroimaging data alone to accurately predict cognitive function. Volumetric T1-weighted neuroimaging data from virally suppressed HIV-positive individuals from the CHARTER cohort (n = 139) were segmented into gray and white matter and spatially normalized before entering into machine learning models. Prediction of cognitive function at baseline and longitudinally was determined using leave-one-out cross-validation. In addition, a multivariate model of brain aging was used to measure the deviation of apparent brain age from chronological age and assess its relationship with cognitive function. Cognitive impairment, defined using the global deficit score, was present in 37.4%. However, it was generally mild and occurred more commonly in those with confounding comorbidities (P < 0.001). Although multivariate prediction of cognitive impairment as a dichotomous variable at baseline was poor (area under the receiver operator curve 0.59), prediction of the global T-score was better than a comparable linear model (adjusted R = 0.08, P < 0.01 vs. adjusted R = 0.01, P = 0.14). Accurate prediction of longitudinal changes in cognitive function was not possible (P = 0.82). Brain-predicted age exceeded chronological age by mean (95% confidence interval) 1.17 (-0.14 to 2.53) years but was greatest in those with confounding comorbidities [5.87 (1.74 to 9.99) years] and prior AIDS [3.03 (0.00 to 6.06) years]. Accurate prediction of cognitive impairment using multivariate models using only T1-weighted data was not achievable, which may reflect the small sample size, heterogeneity of the data, or that impairment was usually mild.


A mechanosensitive caveolae-invadosome interplay drives matrix remodelling for cancer cell invasion.
Invadosomes and caveolae are mechanosensitive structures that are implicated in metastasis. Here, we describe a unique juxtaposition of caveola clusters and matrix degradative invadosomes at contact sites between the plasma membrane of cancer cells and constricting fibrils both in 2D and 3D type I collagen matrix environments. Preferential association between caveolae and straight segments of the fibrils, and between invadosomes and bent segments of the fibrils, was observed along with matrix remodelling. Caveola recruitment precedes and is required for invadosome formation and activity. Reciprocally, invadosome disruption results in the accumulation of fibril-associated caveolae. Moreover, caveolae and the collagen receptor β1 integrin co-localize at contact sites with the fibrils, and integrins control caveola recruitment to fibrils. In turn, caveolae mediate the clearance of β1 integrin and collagen uptake in an invadosome-dependent and collagen-cleavage-dependent mechanism. Our data reveal a reciprocal interplay between caveolae and invadosomes that coordinates adhesion to and proteolytic remodelling of confining fibrils to support tumour cell dissemination.


MYB is essential for mammary tumorigenesis.
MYB oncogene upregulation is associated with estrogen receptor (ER)-positive breast cancer, but disease requirements for MYB function in vivo have not been explored. In this study, we provide evidence of a critical requirement for MYB functions in models of human and murine breast cancer. In human breast cancer, we found that MYB expression was critical for tumor cell growth both in vitro and in vivo in xenograft settings. In transgenic knockout mice, tissue-specific deletion of the murine MYB gene caused a transient defect in mammary gland development that was reflected in delayed ductal branching and defective apical bud formation. In mouse mammary tumor virus (MMTV)-NEU mice where tumors are initiated by activation of HER2, MYB deletion was sufficient to abolish tumor formation. In the more aggressive MMTV-PyMT model system, MYB deletion delayed tumorigenesis significantly. Together, the findings in these transgenic knockout models implied that MYB was critical during an early window in mammary development when it was essential for tumor initiation, even though MYB loss did not exert a lasting impact upon normal mammary function. Two important MYB-target genes that promote cell survival, BCL2 and GRP78/BIP, were each elevated compared with nontransformed mammary epithelial cells, thereby promoting survival as confirmed in colony formation assays in vitro. Taken together, our findings establish a role for MYB at the hub of ER- and HER2-dependent pathways in mammary carcinogenesis.


[Current role of protatic specific antigen (PSA) and its by-products in the diagnosis of prostate cancer].
Incorporation of prostatic specific antigen (PSA) to clinical practice was a revolution in the diagnosis and modified the epidemiology of prostate cancer (PCa). Although it lacks of many characteristics of an ideal tumor marker, it is the marker most used for diagnosis and follow up of any kind of cancer. It represents the best clinical tool we have available today for screening and staging of PCa. On the contrary, its greatest limitation is the lack of tumor specificity. The use of PSA by-products and molecular isoforms tries to solve, at least partially, its limitations. Indeed, the use of FreePSA ratio (%fPSA) ad PSA density (PSAD) increase significantly the specificity of the diagnostic test and, the use of derivatives that evaluate time kinetics of PSA (PSA velocity (PSAV) and PSA doubling time (PSADT) represents a very useful tool for prognosis estimation during treatment and follow up of the disease. The greatest advance over the last years comes from the analysis of the predecessor isoform (-2) pPSA and the phi Index. Both markers have demonstrated to improve the sensitivity and specificity results obtained to date, resulting in a decrease of unnecessary biopsies. Probably, with the ongoing development of new markers for PCa , the role of PSA on disease diagnosis and staging would be modified in a few years.


Density functional theory study of NO adsorbed in A-zeolite.
Density functional theory was employed to investigate the adsorption site and hyperfine interactions of nitric oxide adsorbed in Na-LTA (previous name NaA) zeolite. Three different cluster models of increasing complexity were used to represent the zeolite network: (1) a six-membered ring terminated by hydrogen atoms with one sodium ion above the ring, (2) as model 1 with the addition of three sodium ions located at the centers of three imagined four-membered rings adjacent to the six-membered ring, and (3) as model 2 with the addition of the three four-membered rings adjacent to the six-membered ring. Calculations on the largest system (model 3) showed very good agreement with measured electronic Zeeman interaction couplings, 14N hyperfine coupling tensors, and 23Na hyperfine and nuclear quadruple coupling tensors of the S = 1/2 Na+...N-O adsorption complex when the position of the sodium ion was relaxed. The optimized geometry of the complex agreed nicely with that estimated experimentally, except for the Na-N distance, where the present results indicate that the distance deduced from previous ENDOR experiments may be underestimated by as much as 0.5 angstroms.


PML-nuclear bodies are involved in cellular serum response.
Serum stimulation leads to the activation of various signal transduction pathways in cells, and the resultant signals are integrated into the serum response factor (SRF)-dependent transcription of immediate-early genes such as c-fos. To further characterize this response, we investigated the mechanism which controls serum response transcription in cultured human cells. Frequency of PML (promyelocytic leukaemia)-nuclear bodies (NBs) formation increases shortly after serum stimulation, probably facilitating the interaction of SRF and CBP acetyltransferase at the NBs. PML modulates SRF-mediated c-fos promoter activities upon addition of serum to cells or expression of constitutively active Rho family GTPases. We mapped the region in the SRF that interacts with PML to the C-terminal transactivation domain. An SRF mutant deleted of the transactivation domain neither co-localizes with CBP in NBs nor fulfills its transcriptional role. Under conditions of serum stimulation, the formation of NBs coincides with the immediate-early expression of the endogenous c-fos gene in fibroblasts and in all-trans retinoic acid-treated acute promyelocytic leukaemia NB4 cells. These data provide an insight into the involvement of NBs in modulating the transcription of serum-induced immediate-early genes.


Improved controlled atmosphere high temperature scanning probe microscope.
To locally access electrochemical active surfaces and interfaces in operando at the sub-micron scale at high temperatures in a reactive gas atmosphere is of great importance to understand the basic mechanisms in new functional materials, for instance, for energy technologies, such as solid oxide fuel cells and electrolyzer cells. Here, we report on advanced improvements of our original controlled atmosphere high temperature scanning probe microscope, CAHT-SPM. The new microscope can employ a broad range of the scanning probe techniques including tapping mode, scanning tunneling microscopy, scanning tunneling spectroscopy, conductive atomic force microscopy, and Kelvin probe force microscopy. The temperature of the sample can be as high as 850 °C. Both reducing and oxidizing gases such as oxygen, hydrogen, and nitrogen can be added in the sample chamber and the oxygen partial pressure (pO2) is monitored by an oxygen sensor. We present here some examples of its capabilities demonstrated by high temperature topography with simultaneously ac electrical conductance measurements during atmosphere changes, electrochemical impedance spectroscopy at various temperatures, and measurements of the surface potential. The improved CAHT-SPM, therefore, holds a great potential for local sub-micron analysis of high-temperature and gas induced changes of a wide range of materials.


Long-term effectiveness of adjuvant goserelin in premenopausal women with early breast cancer.
Systematic reviews have found that luteinizing hormone-releasing hormone (LHRH) agonists are effective in treating premenopausal women with early breast cancer. We conducted long-term follow-up (median 12 years) of 2706 women in the Zoladex In Premenopausal Patients (ZIPP), which evaluated the LHRH agonist goserelin (3.6 mg injection every 4 weeks) and tamoxifen (20 or 40 mg daily), given for 2 years. Women were randomly assigned to receive each therapy alone, both, or neither, after primary therapy (surgery with or without radiotherapy/chemotherapy). Hazard ratios and absolute risk differences were used to assess the effect of goserelin treatment on event-free survival (breast cancer recurrence, new tumor or death), overall survival, risk of recurrence of breast cancer, and risk of dying from breast cancer, in the presence or absence of tamoxifen. Fifteen years after the initiation of treatment, for every 100 women not given tamoxifen, there were 13.9 (95% confidence interval [CI] = 17.5 to 19.4) fewer events among those who were treated with goserelin compared with those who were not treated with goserelin. However, among women who did take tamoxifen, there were 2.8 fewer events (95% CI = 7.7 fewer to 2.0 more) per 100 women treated with goserelin compared with those not treated with goserelin. The risk of dying from breast cancer was also reduced at 15 years: For every 100 women given goserelin, the number of breast cancer deaths was lower by 2.6 (95% CI = 6.6 fewer to 2.1 more) and 8.5 (95% CI = 2.2 to 13.7) in those who did and did not take tamoxifen, respectively, although in the former group the difference was not statistically significant. Two years of goserelin treatment was as effective as 2 years of tamoxifen treatment 15 years after starting therapy. In women who did not take tamoxifen, there was a large benefit of goserelin treatment on survival and recurrence, and in women who did take tamoxifen, there was a marginal potential benefit on these outcomes when goserelin was added.


Evidence that tenascin and thrombospondin-1 modulate sprouting of endothelial cells.
Cultured endothelial cells undergo a reversible transition from a resting (cobblestone) phenotype to an angiogenic (sprouting) phenotype. This transition mimics the early events of angiogenesis. We have previously reported that the addition of exogenous xylosides inhibits endothelial cel sprouting and modifies the extracellular matrix (ECM) synthesised by the cells. We have now investigated whether endothelial sprouting is mediated by the nature of the extracellular matrix in contact with the cells. Accordingly, cell-free matrices deposited by bovine aortic endothelial cells (BAEC) were isolated. These matrices were produced under conditions in which the formation of the sprouting phenotype was permitted (controls) or inhibited (by the addition of exogenous xylosides). BAEC were then plated on these matrices and grown under conditions which promote sprouting. Sprouting proceeded normally on control matrices, whereas it was inhibited when the cells were grown on matrices deposited in the presence of xylosides. The composition of the permissive and inhibitory matrices was then analysed. Inhibitory matrices contained reduced levels of tenascin and increased levels of thrombospondin-1 by comparison to the permissive matrices. In contrast, no differences were detected in the relative levels of laminin. The roles of tenascin and thrombospondin-1 in endothelial sprouting were confirmed using specific antibodies. Immunolocalisation studies revealed the presence of both proteins in sprouting cells. Antibodies to tenascin inhibited the formation of sprouting cells on permissive matrices and on gelatin-coated dishes without affecting cell growth. Tenascin synthesis was increased when sprouting cells were present in the cultures. Antibodies to thrombospondin-1 stimulated sprouting on inhibitory matrices. These results suggest that the transition from a resting to a sprouting phenotype is promoted by tenascin and inhibited by thrombospondin-1.


Low intensity blood flow restriction training: a meta-analysis.
The primary objective of this investigation was to quantitatively identify which training variables result in the greatest strength and hypertrophy outcomes with lower body low intensity training with blood flow restriction (LI-BFR). Searches were performed for published studies with certain criteria. First, the primary focus of the study must have compared the effects of low intensity endurance or resistance training alone to low intensity exercise with some form of blood flow restriction. Second, subject populations had to have similar baseline characteristics so that valid outcome measures could be made. Finally, outcome measures had to include at least one measure of muscle hypertrophy. All studies included in the analysis utilized MRI except for two which reported changes via ultrasound. The mean overall effect size (ES) for muscle strength for LI-BFR was 0.58 [95% CI: 0.40, 0.76], and 0.00 [95% CI: -0.18, 0.17] for low intensity training. The mean overall ES for muscle hypertrophy for LI-BFR training was 0.39 [95% CI: 0.35, 0.43], and -0.01 [95% CI: -0.05, 0.03] for low intensity training. Blood flow restriction resulted in significantly greater gains in strength and hypertrophy when performed with resistance training than with walking. In addition, performing LI-BFR 2-3 days per week resulted in the greatest ES compared to 4-5 days per week. Significant correlations were found between ES for strength development and weeks of duration, but not for muscle hypertrophy. This meta-analysis provides insight into the impact of different variables on muscular strength and hypertrophy to LI-BFR training.


Pharmacoresistant partial-onset epilepsy misdiagnosed as panic disorder: a case report.
The clinical differentiation between simple partial epileptic seizures of temporal lobe origin and panic attacks is often difficult on clinical grounds alone, because both conditions are characterized by common symptomatology which includes the feeling of fear, autonomic system dysfunction, disorientation and alternation of the level of consciousness when these conditions evolve clinically. The symptoms stem from common pathophysiologic and anatomic substrates of these two conditions, localized in the limbic system, especially the amygdala. We present the case of a young woman who had a febrile seizure in childhood and subsequent episodes of fear accompanied by tremor and possible alteration of consciousness followed by headache. These spells were diagnosed as panic attacks during her teenage years and she was given Clobazam in order to suppress them. The patient responded well for several years without attacks but her symptoms reappeared following discontinuation of her medication in order to conceive. At that time a detailed history was taken from her spouse and further clinical evaluation raised the suspicion of seizures especially due to the fact that her spells were characterized by alteration of consciousness; she was therefore referred for additional investigations which included admission to a monitoring unit for epilepsy. Long-term video-EEG recording revealed the presence of simple partial seizures with secondary generalization confirming the clinical impression. She was subsequently treated with antiepileptic medications; however the patient's condition worsened to the point where she became pharmacoresistant having failed several antiepileptic drug trials in monotherapy or combination. An MRI scan of the brain revealed the presence of right-sided mesial temporal sclerosis, a known consequence of febrile seizures. Her seizures were nocturnal tonic-clonic and gradually worsened to the point of occurring during most nights. She was therefore referred for a presurgical evaluation which confirmed that the epileptic focus was associated with the area of mesial temporal sclerosis. The epileptic focus was successfully removed from the right anterior temporal lobe and since then she remained free of seizures whereas, in addition, the presumed symptoms of panic attacks also resolved. This case indicates the occasional difficulty in diagnosing simple partial seizures and how it may be confused with psychiatric conditions. Therefore, the treating physician, especially the psychiatrist, should remain vigilant when treating cases of panic attacks, especially when they present with either atypical symptomatology, such as the case described, or when they do not respond to appropriately chosen treatment; such cases may warrant referral for further investigation.


Efficacy of brodalumab in the treatment of scalp and nail psoriasis: results from three phase 3 trials.
We evaluated the efficacy of brodalumab in patients with nail or scalp psoriasis in three phase 3 studies (AMAGINE-1/-2/-3). In AMAGINE-1, scalp clearance, measured by the psoriasis scalp severity index (PSSI), was reported for patients who received brodalumab 210 mg every 2 weeks (Q2W) or placebo through 12 weeks. In AMAGINE-2/-3, nail clearance, measured by the nail psoriasis severity index (NAPSI), was reported for patients receiving either brodalumab 210 mg Q2W or ustekinumab continuously through 52 weeks. At week 12, significantly more patients receiving brodalumab achieved 75% and 100% improvement rates from baseline PSSI and had lower mean PSSI across 12 weeks compared with placebo, with significant improvement in PSSI evident with brodalumab 210 mg Q2W vs placebo by week 2. Across 52 weeks, patients receiving brodalumab achieved significantly greater complete clearance of nail psoriasis (NAPSI 0), lower mean NAPSI, and higher mean percent improvement rates from baseline NAPSI than patients receiving ustekinumab. At week 52, 63.8% of patients receiving brodalumab achieved NAPSI 0 vs 39.1% of patients receiving ustekinumab. Brodalumab was associated with clearance of scalp psoriasis through 12 weeks and improvements in nail psoriasis, including complete nail clearance, through 52 weeks.


Recent advances in Barrett's esophagus.
Barrett's esophagus (BE) is the only known precursor of esophageal adenocarcinoma, one of the few cancers with increasing incidence in developed countries. The pathogenesis of BE is unclear with regard to either the cellular origin of this metaplastic epithelium or the manner in which malignant transformation occurs, although recent data indicate a possible junctional origin of stem cells for BE. Treatment of BE may be achieved using endoscopic eradication therapy; however, there is a lack of discriminatory tools to identify individuals at sufficient risk for cancer development in whom intervention is warranted. Reduction in gastroesophageal reflux of gastric contents including acid is mandatory to achieve remission from BE after endoscopic ablation, and can be achieved using medical or nonmedical interventions. Research topics of greatest interest include the mechanism of BE development and transformation to cancer, risk stratification methods to identify individuals who may benefit from ablation of BE, optimization of eradication therapy, and surveillance methods to ensure that remission is maintained after eradication is achieved.


A cost of illness study of hypoglycaemic events in insulin-treated diabetes in the Netherlands.
Patients with diabetes mellitus are at a risk for hypoglycaemia. Besides the burden of hypoglycaemia for patients, hypoglycaemia poses an economic burden to society. The aim of this study was to calculate the per patient societal costs of hypoglycaemia among patients with type1 diabetes (T1DM) and type 2 diabetes (T2DM) on insulin therapy in the Netherlands. To calculate the costs of hypoglycaemia, data from the Global Hypoglycaemia Assessment Tool (HAT) study were used. Dutch patients were selected from the HAT study database and data regarding healthcare resource use, informal care use and productivity losses were combined with Dutch unit costs to calculate the per patient 4-week costs of patients experiencing hypoglycaemia. Besides these 4-week costs, costs per hypoglycaemic event were calculated by dividing the study population total 4-week costs by the total number of events in this period. Mean 4-week total costs of hypoglycaemia amounted to €163 (SD, €870) in T1DM and €134 (SD, €364) in T2DM. While productivity costs were the most important cost driver of hypoglycaemia in patients with T1DM (accounting for 72% of the total costs), costs of hypoglycaemia in patients with T2DM were almost entirely driven by costs within the healthcare sector (accounting for 98% of the total costs). Mean costs of a severe hypoglycaemic event were €828 and €508 in T1DM and T2DM, respectively, whereas mean costs of a non-severe event were almost zero. This study showed that the economic burden of severe hypoglycaemia is substantial. The prevention of hypoglycaemia could therefore not only reduce the burden for patients, but also the economic burden to society.


A Comparison of Mixed-Method Cooling Interventions on Preloaded Running Performance in the Heat.
Stevens, CJ, Bennett, KJM, Sculley, DV, Callister, R, Taylor, L, and Dascombe, BJ. A comparison of mixed-method cooling interventions on preloaded running performance in the heat. J Strength Cond Res 31(3): 620-629, 2017-The purpose of this investigation was to assess the effect of combining practical methods to cool the body on endurance running performance and physiology in the heat. Eleven trained male runners completed 4 randomized, preloaded running time trials (20 minutes at 70% V[Combining Dot Above]O2max and a 3 km time trial) on a nonmotorized treadmill in the heat (33° C). Trials consisted of precooling by combined cold-water immersion and ice slurry ingestion (PRE), midcooling by combined facial water spray and menthol mouth rinse (MID), a combination of all methods (ALL), and control (CON). Performance time was significantly faster in MID (13.7 ± 1.2 minutes; p < 0.01) and ALL (13.7 ± 1.4 minutes; p = 0.04) but not PRE (13.9 ± 1.4 minutes; p = 0.24) when compared with CON (14.2 ± 1.2 minutes). Precooling significantly reduced rectal temperature (initially by 0.5 ± 0.2° C), mean skin temperature, heart rate and sweat rate, and increased iEMG activity, whereas midcooling significantly increased expired air volume and respiratory exchange ratio compared with control. Significant decreases in forehead temperature, thermal sensation, and postexercise blood prolactin concentration were observed in all conditions compared with control. Performance was improved with midcooling, whereas precooling had little or no influence. Midcooling may have improved performance through an attenuated inhibitory psychophysiological and endocrine response to the heat.


Perforin deficiency attenuates collagen-induced arthritis.
Collagen-induced arthritis (CIA), an approved animal model for rheumatoid arthritis, is thought to be a T cell-dependent disease. There is evidence that CD8+ T cells are a major subset controlling the pathogenesis of CIA. They probably contribute to certain features of disease, namely tissue destruction and synovial hyperplasia. In this study we examined the role of perforin (pfp), a key molecule of the cytotoxic death pathway that is expressed mainly in CD8+ T cells, for the pathogenesis of CIA. We generated DBA/1J mice suffering from mutations of the pfp molecule, DBA/1J-pfp-/-, and studied their susceptibility to arthritis. As a result, pfp-deficient mice showed a reduced incidence (DBA/1J-pfp+/+, 64%; DBA/1J-pfp-/-, 54%), a slightly delayed onset (onset of disease: DBA/1J-pfp+/+, 53 +/- 3.6; DBA/1J-pfp-/-, 59 +/- 4.9 (mean +/- SEM), and milder form of the disease (maximum disease score: DBA/1J-pfp+/+, 7.3 +/- 1.1; DBA/1J-pfp-/-, 3.4 +/- 1.4 (mean +/- SEM); P < 0.05). Concomitantly, peripheral T cell proliferation in response to the specific antigen bovine collagen II was increased in pfp-/- mice compared with pfp+/+ mice, arguing for an impaired killing of autoreactive T cells caused by pfp deficiency. Thus, pfp-mediated cytotoxicity is involved in the initiation of tissue damage in arthritis, but pfp-independent cytotoxic death pathways might also contribute to CIA.


Demographic and genetic structures of two partially isolated communities of Santa Catarina Island, southern Brazil.
The objectives of this study were to analyze the population structure and genetic variability of two communities, Costa da Lagoa (CLG) and São João do Rio Vermelho (SJRV), located on Santa Catarina Island in southern Brazil. The two populations descend from Azores Archipelago immigrants (Portuguese), with a minor contribution of sub-Saharan Africans and Amerindians. To estimate the relative contribution of the different ethnic groups to the current gene pool of the two communities, values of admixture were obtained using the weighted least-squares method based on allelic frequencies of the loci ABO, RHD-RHCE, GPA-GPB (MNSs), HBB, HP, TF, CP, AK, and ACP1. The origins of the studied populations can be quantified as follows: for CLG, sub-Saharan Africans (A) = 17.3%, Iberian Europeans (P) = 75.0%, and Southern Amerindians (I) = 7.7%; for SJRV, A = 48.8%, P = 44.5%, and I = 6.7%. Because haplotype frequencies of the GPA-GPB loci in SJRV were unusual, possibly as a consequence of random genetic drift, the values of admixture were recalculated after exclusion of GPA-GPB, as follows: A = 28.0%; P = 53.3%, and I = 18.7%. The total diversity (HT) was estimated as 42.29%, of which 99.6% can be attributed to the intrapopulational variability (HS). The interpopulational genetic variation (or standard distance, DST) corresponds to 0.19%, while the gene differentiation coefficient is 0.28%, indicative of low genetic difference. These results led to the conclusion that random genetic drift may have had an important effect on the Costa da Lagoa community, while presently gene flow might be the predominant evolutionary factor potentially capable of changing allele frequencies in SJRV.


Mass-density compensation can improve the performance of a range of different detectors under non-equilibrium conditions.
Dosimeters often consist of several components whose mass densities differ substantially from water. These components cause small-field correction factors to vary significantly as lateral electronic equilibrium breaks down. Even amongst instruments designed for small-field dosimetry, inter-detector variation in the correction factors associated with very small (∼0.5 cm) fields can amount to tens of per cent. For a given dosimeter, small-field correction factors vary not only with field size but also with detector azimuthal angle and position within the field. Furthermore the accurate determination of these factors typically requires time-intensive Monte Carlo simulations. Thus, if achievable, 'correction factor free' small-field dosimetry would be highly desirable. This study demonstrates that a new generation of mass-density compensated detectors could take us towards this goal. Using a 6 MV beam model, it shows that 'mass-density compensation' can be utilized to improve the performance of a range of different detectors under small-field conditions. Non-sensitive material of appropriate mass-density is incorporated into detector designs in order to make the instruments behave as if consisting only of water. The dosimeter perturbative effects are then reduced to those associated with volume averaging. An even better solution-which modifies detectors to obtain profiles that look like those measured by a point-like water structure-is also considered. Provided that adequate sensitivity can be achieved for a small measurement volume, this study shows that it may be possible to use mass-density compensation (and Monte Carlo-driven design) to produce a solid-state dosimeter/ionization chamber with a near-perfect non-equilibrium response.


Evaluation of High-Temperature Hydrogen Sensors Based on BaCe0.6Zr0.3Y0.1O3-α and Sr(Ce0.9Zr0.1)0.95Yb0.05O3-α Perovskites for Industrial Applications.
In many industrial fields, there is a need to design and characterize on-line and on-board hydrogen monitoring tools able to operate under extreme conditions. One of these applications is in future nuclear fusion reactors, which will use hydrogen isotopes as a plasma fuel. In this context, the measurement of the concentration of these hydrogen isotopes will be of interest to ensure the correct operating conditions for such reactors. Hydrogen sensors based on solid-state electrolytes will be the first step in the development of new analytical tools able to quantify deuterium and tritium in aggressive environments. In the present work, amperometric hydrogen sensors were constructed and evaluated using two solid-state electrolytes, BaCe0.6Zr0.3Y0.1O3-α and Sr(Ce0.9Zr0.1)0.95Yb0.05O3-α. Prototype sensors were built in order to study their sensitivity in on-line measurements. The experiments were performed in a reactor with a hydrogen-controlled environment. The sensors were evaluated at 500 and 600 °C in amperometric mode by applying 2 and 4 V voltages between electrodes. Both sensors showed increases in sensitivity when the temperature or voltage were increased.


The antidiabetic and anticholinergic effects of chrysin on cyclophosphamide-induced multiple organ toxicity in rats: Pharmacological evaluation of some metabolic enzyme activities.
Chrysin (CH) or 5,7-dihydroxyflavone is a flavonoid present in various plants, bee propolis, and honey. Cyclophosphamide (CYP) is a chemotherapeutic drug, which is extensively used in the treatment of multiple human malignancies. In our study, we aimed to investigate the effects of CYP and CH on some metabolic enzymes including carbonic anhydrase, aldose reductase, paraoxonase-1, α-glycosidase, acetylcholinesterase, and butyrylcholinesterase enzyme activities in the brain, heart, testis, liver, and kidney tissues of rats. Thirty-five adult male Wistar rats were used. The animals were pretreated with CH (25 and 50 mg/kg b.w.) for seven days before administering a single dose of CYP (200 mg/kg b.w.) on the seventh day. In all the tissues, the treatment of CH significantly regulated these enzyme activities in CYP-induced rats. These results showed that CH exhibited an ameliorative effect against CYP-induced brain, heart, liver, testis, and kidney toxicity.


A pragmatic, multicentre, randomised controlled trial comparing stapled haemorrhoidopexy to traditional excisional surgery for haemorrhoidal disease (eTHoS): study protocol for a randomised controlled trial.
Current interventions for haemorrhoidal disease include traditional haemorrhoidectomy (TH) and stapled haemorrhoidopexy (SH) surgery. However, uncertainty remains as to how they compare from a clinical, quality of life (QoL) and economic perspective. The study is therefore designed to determine whether SH is more effective and more cost-effective, compared with TH. eTHoS (either Traditional Haemorrhoidectomy or Stapled Haemorrhoidopexy for Haemorrhoidal Disease) is a pragmatic, multicentre, randomised controlled trial. Currently, 29 secondary care centres are open to recruitment. Patients, aged 18 year or older, with circumferential haemorrhoids grade II to IV, are eligible to take part. The primary clinical and economic outcomes are QoL profile (area under the curve derived from the EuroQol Group's 5 Dimension Health Status Questionnaire (EQ-5D) at all assessment points) and incremental cost per quality adjusted life year (QALY) based on the responses to the EQ-5D at 24 months. The secondary outcomes include a comparison of the SF-36 scores, pain and symptoms sub-domains, disease recurrence, complication rates and direct and indirect costs to the National Health Service (NHS). A sample size of n =338 per group has been calculated to provide 90% power to detect a difference in the mean area under the curve (AUC) of 0.25 standard deviations derived from EQ-5D score measurements, with a two-sided significance level of 5%. Allowing for non-response, 400 participants will be randomised per group. Randomisation will utilise a minimisation algorithm that incorporates centre, grade of haemorrhoidal disease, baseline EQ-5D score and gender. Blinding of participants and outcome assessors is not attempted. This is one of the largest trials of its kind. In the United Kingdom alone, 29,000 operations for haemorrhoidal disease are done annually. The trial is therefore designed to give robust evidence on which clinicians and health service managers can base management decisions and, more importantly, patients can make informed choices. Current Controlled Trials ISRCTN80061723 (assigned 8 March 2010).


Short communication: Herd-level variables associated with overmilking in Michigan dairy herds.
The objective of this study was to determine the herd-level variables that are associated with overmilking in 64 Michigan dairy herds with a mean herd size of 451 cows (range: 59-2,771 cows). Participating producers completed surveys to indicate their mastitis management practices and attitudes. Additionally, milking protocols were observed and milk flow dynamics for 3,824 cows were estimated using digital vacuum recorders. The median duration of overmilking was 47 s (95% confidence interval, CI: 38.6 to 55.9 s), with a mean of 55% (95% CI: 49.5 to 61.1%) of cows within each herd overmilked by at least 30 s. Median milking time for all herds was 324 s (95% CI: 302 to 346 s) and was found to be positively correlated with median duration of overmilking (r = 0.670). Backward multivariate analysis was used to determine which of 45 herd-level milking and management variables were associated with median duration of overmilking. Median duration of overmilking was negatively associated with the duration of time needed to complete 1 milking for the entire herd (adjusted R2 = 0.13). Herds that operate milking facilities below maximum daily capacity may be prone to overmilking. Given the low coefficient of determination, variables unaccounted for in this study, such as equipment function or manual detachment by milking operators, are likely the most important risk factors for overmilking.


Phylogeny and expression of carbonic anhydrase-related proteins.
Carbonic anhydrases (CAs) are found in many organisms, in which they contribute to several important biological processes. The vertebrate alpha-CA family consists of 16 subfamilies, three of which (VIII, X and XI) consist of acatalytic proteins. These are named carbonic anhydrase related proteins (CARPs), and their inactivity is due to absence of one or more Zn-binding histidine residues. In this study, we analyzed and evaluated the distribution of genes encoding CARPs in different organisms using bioinformatic methods, and studied their expression in mouse tissues using immunohistochemistry and real-time quantitative PCR. We collected 84 sequences, of which 22 came from novel or improved gene models which we created from genome data. The distribution of CARP VIII covers vertebrates and deuterostomes, and CARP X appears to be universal in the animal kingdom. CA10-like genes have had a separate history of duplications in the tetrapod and fish lineages. Our phylogenetic analysis showed that duplication of CA10 into CA11 has occurred only in tetrapods (found in mammals, frogs, and lizards), whereas an independent duplication of CA10 was found in fishes. We suggest the name CA10b for the second fish isoform. Immunohistochemical analysis showed a high expression level of CARP VIII in the mouse cerebellum, cerebrum, and also moderate expression in the lung, liver, salivary gland, and stomach. These results also demonstrated low expression in the colon, kidney, and Langerhans islets. CARP X was moderately expressed in the cerebral capillaries and the lung and very weakly in the stomach and heart. Positive signals for CARP XI were observed in the cerebellum, cerebrum, liver, stomach, small intestine, colon, kidney, and testis. In addition, the results of real-time quantitative PCR confirmed a wide distribution for the Car8 and Car11 mRNAs, whereas the expression of the Car10 mRNA was restricted to the frontal cortex, parietal cortex, cerebellum, midbrain, and eye. CARP sequences have been strongly conserved between different species, and all three CARPs show high expression in the mouse brain and CARP VIII is also expressed in several other tissues. These findings suggest an important functional role for these proteins in mammals.


Early Blood Transfusions in Sepsis: Unchanged Survival and Increased Costs.
Early red blood cell transfusions are a common treatment for adults hospitalized for sepsis without shock. However, their utility and association with mortality and costs have not been well studied. To examine early transfusion rates for patients with sepsis treated outside intensive care units, and to find a correlation between transfusion rates and survival rates and costs. Data were obtained from hospital members of the Premier Healthcare Alliance that admitted at least 50 adults with sepsis between January 1, 2006, and December 31, 2010. Early transfusion rates at each hospital were calculated as the observed incidence of allogeneic red blood cells administered by hospital day 2. A multivariable logistic regression model was constructed to estimate the expected or risk-adjusted transfusion rates, mortality rates, and costs. A total of 256 396 adults were hospitalized with sepsis without major bleeding or surgery at 364 US hospitals. Approximately 84% of all patients admitted with sepsis, without vasopressor therapy, were treated outside the intensive care unit (by day 2). The mean institutional early transfusion rate was 6.9%. After risk standardization, the median (interquartile range) transfusion rate was 6.7% (5.8%-7.6%), mortality rate was 15.5% (13.1%-18.1%), and costs were $13 333 ($11 939-$14 986). Early transfusion rates were not correlated with mortality but were modestly positively correlated with costs (Spearman ρ = 0.157; P = .003). Early transfusion rates during hospitalization for sepsis without shock varied widely across the hospitals. Transfusion rates were associated with increased costs but not with mortality rates.