Women smokers' experiences of an age-appearance anti-smoking intervention: a qualitative study.
This study was designed to investigate women's experiences of engaging in an age-appearance anti-smoking intervention. Ten 18- to 34-year-old women gave accounts of their experiences after engaging in an age-appearance facial morphing anti-smoking intervention in interviews (n= 7) and a focus group (n= 3), and 37 women gave their accounts while they were engaged in the intervention. Transcripts were analysed using a thematic analysis broadly informed by the procedures of Grounded Theory. Women were very concerned about the impact of ageing on their faces in general, and in particular the additional impact of smoking on their skin. Women were concerned about other people's reactions to them as older smokers with wrinkled skin, and many experienced a physical shock reaction (including reports of nausea) to seeing how they would age if they continued to smoke. They reported that seeing their own face aged on the computer screen increased their perceived risk of skin wrinkling. Women reported being highly motivated to quit smoking as a result of the intervention, and many reported that they would take active steps to quit having seen how they would look if they continued to smoke. This was linked with increased perceived personal responsibility for quitting. Results are discussed in relation to suggestions for anti-smoking interventions aimed at women in the 18- to 34-year-old age group. It is concluded that interventions incorporating age-appearance morphing techniques are likely to be effective in helping women to take active steps to quit smoking.


No association between hemoglobin A1c and in-hospital mortality in patients with diabetes and acute myocardial infarction.
Patients with diabetes have increased in-hospital mortality following acute myocardial infarction (AMI), with studies suggesting higher risk with both hypoglycemia and hyperglycemia. We assessed whether a J-shaped relation exists between hemoglobin A1c (A1C) in patients with diabetes and AMI. We assessed the associations between A1C and in-hospital mortality using data from a nationwide sample of AMI patients who had both prior diabetes and measurement of A1C (N = 15,337). When evaluated continuously, we observed no evidence of a J-shaped relation between A1C and in-hospital mortality in multivariable analysis (test for linearity P = .89). Patients with lowest (<5.5%) and highest A1C (≥9.5%) had a crude mortality rate of 4.6% and 2.8%, respectively, compared with 3.8% among those in the referent A1C category (6.5% to <7%). In multivariable regression, we observed no association between low A1C (<5.5%, odds ratio 0.81, 95% CI 0.47-1.39) or high A1C (A1C ≥9.5, odds ratio 1.31, 95% CI 0.94-1.83) and mortality as compared with the referent group. These findings can only be generalized to the subset of patients with diabetes who had A1C assessed during their hospitalization; these patients tended to be healthier than those in whom A1C was not assessed. In this large contemporary cohort of patients with diabetes presenting with AMI, we did not observe a J-shaped association between A1C and mortality.


Relationship of coping style to mood and anxiety disorders in dementia carers.
Mood and anxiety disorders are common in family carers of people with dementia, and are associated with poor carer and care recipient outcomes. Caring for someone with dementia is stressful, but how carers cope may be an important determinant of carer psychological morbidity. This review summarizes and integrates the literature on the effect of coping style and its role in effective carer interventions. Two systematic reviews evaluated relationships between coping style and mood/anxiety disorders in observational studies and randomized controlled trials (RCTs), respectively. Several subsequent cross-sectional studies have also investigated these relationships, including in the context of coping as a mediator or moderator. Studies also investigated suicidality and cultural factors underlying coping. More dysfunctional, and less emotional support and acceptance-based coping styles are robustly associated with carer depression and anxiety symptoms and caseness in cross-sectional studies. There is also preliminary evidence from longitudinal studies that these coping styles may predict subsequent carer psychological morbidity. Psychological interventions successful in reducing depression counterintuitively increased dysfunctional coping in RCTs, but no RCTs have directly tested whether emotional support and acceptance-based coping mediates the effects of psychological intervention.


Higher and increased concentration of hs-CRP within normal range can predict the incidence of metabolic syndrome in healthy men.
To determine whether hs-CRP level and interval change of hs-CRP could predict the development of metabolic abnormalities in healthy subjects in a longitudinal study. A cohort of 3748 male who had normal hs-CRP level without evidence of any component of metabolic syndrome were studied. At each visit, hs-CRP level and metabolic abnormalities were measured. Interval change of hs-CRP for each subject was calculated. COX proportional hazard model and logistic regression analysis were used. Cumulative incidence and incidence density of metabolic syndrome were 3.96% and 7.17 per 1000 person-year, respectively. Cumulative incidence of metabolic syndrome was significantly increased according to hs-CRP tertile level. This significance remained after adjusting age, smoking, drinking, and exercise. Although the hazard ratio of metabolic syndrome for incidence density was increased significantly as hs-CRP increased, such trend disappeared after adjusting for confounding variables. The risk of metabolic syndrome was significantly higher (1.48 times) in the hs-CRP increased group than that in the decreased or unchanged group. This significance remained after controlling for covariates. Relatively higher hs-CRP level within normal range may predict the increase of metabolic syndrome compared to lower hs-CRP. Increased hs-CRP level may increase the incidence of metabolic syndrome.


Effect of rosmarinic acid and carnosic acid on AGEs formation in vitro.
This work aimed to investigate the effect of the two main components of rosemary extracts, namely rosmarinic acid (RA) and carnosic acid (CA), on the formation of advanced glycation end-products (AGEs) in vitro. In the bovine serum albumin (BSA)/glucose model, addition of RA and CA at 400μg/mL inhibited fluorescent AGEs by more than 90%, and carboxymethyl lysine (CML) and carboxyethyl lysine (CEL) by 82.7% and 75.2%, and 71.4% and 64.2%, respectively. Moreover, the addition of RA and CA at 400μg/mL inhibited fluorescent AGEs by more than 90% both in the BSA/glyoxal (GO) and BSA/methylglyoxal (MGO) models, the formation of CML by 64.9% and 53.9% in BSA/GO model, and CEL by 28.9% and 24.3% in BSA/MGO model, respectively. RA and CA also significantly decreased the concentration of MGO and protein carbonylation.


Job insecurity and risk of diabetes: a meta-analysis of individual participant data.
Job insecurity has been associated with certain health outcomes. We examined the role of job insecurity as a risk factor for incident diabetes. We used individual participant data from 8 cohort studies identified in 2 open-access data archives and 11 cohort studies participating in the Individual-Participant-Data Meta-analysis in Working Populations Consortium. We calculated study-specific estimates of the association between job insecurity reported at baseline and incident diabetes over the follow-up period. We pooled the estimates in a meta-analysis to produce a summary risk estimate. The 19 studies involved 140 825 participants from Australia, Europe and the United States, with a mean follow-up of 9.4 years and 3954 incident cases of diabetes. In the preliminary analysis adjusted for age and sex, high job insecurity was associated with an increased risk of incident diabetes compared with low job insecurity (adjusted odds ratio [OR] 1.19, 95% confidence interval [CI] 1.09-1.30). In the multivariable-adjusted analysis restricted to 15 studies with baseline data for all covariates (age, sex, socioeconomic status, obesity, physical activity, alcohol and smoking), the association was slightly attenuated (adjusted OR 1.12, 95% CI 1.01-1.24). Heterogeneity between the studies was low to moderate (age- and sex-adjusted model: I2 = 24%, p = 0.2; multivariable-adjusted model: I2 = 27%, p = 0.2). In the multivariable-adjusted analysis restricted to high-quality studies, in which the diabetes diagnosis was ascertained from electronic medical records or clinical examination, the association was similar to that in the main analysis (adjusted OR 1.19, 95% CI 1.04-1.35). Our findings suggest that self-reported job insecurity is associated with a modest increased risk of incident diabetes. Health care personnel should be aware of this association among workers reporting job insecurity.


Relative frequency and morphology of cancers in STK11 mutation carriers.
There is limited data on the spectrum and risk for cancer associated with germline serine/threonine protein kinase 11 (STK11) mutations that cause Peutz-Jeghers syndrome (PJS). We analyzed the incidence of cancer in 240 individuals with PJS possessing germline mutations in STK11. Fifty-four cancers were found among carriers. Overall, the risk for developing cancer at ages 20, 30, 40, 50, 60, and 70 years was 1%, 3%, 19%, 32%, 63%, and 81%, respectively. Kaplan-Meier analysis showed similar cancer risks between missense and truncating mutation carriers (log-rank chi(2) = 2.48; P = 0.12). There was some evidence that mutations in exon 3 of STK11 were associated with a higher cancer risk than mutations within other regions of the gene. We found no difference in overall cancer risk between male and female carriers (log-rank chi(2) = 1.31; P = 0.25) or between familial and sporadic cases (log-rank chi(2) = 1.16, with 1 df; P = 0.28). The most common cancers represented were gastrointestinal in origin--gastroesophageal, small bowel, colorectal, and pancreatic--and the risk for these cancers at ages 30, 40, 50, and 60 years was 1%, 10%, 18%, and 42%, respectively. In women, the risk for breast cancer was substantially increased, being 32% by age 60 years. These results quantitatively show the spectrum of cancer risk associated with STK11 germline mutations in the context of PJS and provide a valuable reference for defining surveillance regimens.


Echinacea purpurea-derived alkylamides exhibit potent anti-inflammatory effects and alleviate clinical symptoms of atopic eczema.
Atopic eczema (AE) is a chronic inflammatory and pruritic skin disease. There is still an unmet need for topical anti-inflammatory and anti-pruritic substances exhibiting an excellent safety profile. The endocannabinoid system is known to regulate various aspects of cutaneous barrier and immune functions, thus targeting it may be a valid approach for alleviating the symptoms of AE. To assess the putative efficacy of Echinacea purpurea-derived alkylamides (Ec. extract) activating cannabinoid (CB)-2 receptors in exerting anti-inflammatory effects and alleviating symptoms of AE. In vitro anti-inflammatory efficiency was investigated by monitoring the effects of Ec. extract on poly-(I:C)-induced pro-inflammatory cytokine expression (Q-PCR) and release (ELISA) of HaCaT keratinocytes. Irritancy and sensitization potential (assessed by Human Repeat Insult Patch Test; Clinical trial 1); clinical efficiency in alleviating symptoms of AE (Clinical trial 2) as well as effects on human skin structure and lipid content (Clinical trial 3 followed by transmission electron microscopy and HPTLC) were investigated in randomized double blind clinical trials. Ec. extract significantly reduced mRNA expression as well as release of poly-(I:C)-induced pro-inflammatory cytokines (IL-6 and IL-8) in keratinocytes. Thus, not surprisingly, the well-tolerated (Clinical trial 1) Ec. extract-based cream reduced local SCORAD statistically significantly, not only compared to baseline, but also compared to the comparator (Clinical trial 2). Of great importance, besides the in vitro anti-inflammatory effects, administration of the Ec. extract-based cream also resulted in significantly higher levels of overall epidermal lipids, ceramide EOS (ω-esterified fatty acid+sphingosine sphingoid base), and cholesterol at Day 15 compared to baseline as well as significantly greater numbers of intercellular lipid lamellae in the intercellular space (Clinical trial 3). The investigated Ec. extract shows great potential in alleviating cutaneous symptoms of AE, and by exerting remarkable anti-inflammatory actions and restoring the epidermal lipid barrier, it will be very likely a well-tolerated, powerful novel ingredient for the adjuvant therapy of AE.


High and low roads to odor valence? A choice response-time study.
Valence and edibility are two important features of olfactory perception, but it remains unclear how they are read out from an olfactory input. For a given odor object (e.g., the smell of rose or garlic), does perceptual identification of that object necessarily precede retrieval of information about its valence and edibility, or alternatively, are these processes independent? In the present study, we studied rapid, binary perceptual decisions regarding odor detection, object identity, valence, and edibility for a set of common odors. We found that decisions regarding odor-object identity were faster than decisions regarding odor valence or edibility, but slower than detection. Mediation analysis revealed that odor valence and edibility decision response times were predicted by a model in which odor-object identity served as a mediator along the perceptual pathway from detection to both valence and edibility. According to this model, odor valence is determined through both a "low road" that bypasses odor objects and a "high road" that utilizes odor-object information. Edibility evaluations are constrained to processing via the high road. The results outline a novel causal framework that explains how major perceptual features might be rapidly extracted from odors through engagement of odor objects early in the processing stream.


Understanding the effects of electromagnetic field emissions from Marine Renewable Energy Devices (MREDs) on the commercially important edible crab, Cancer pagurus (L.).
The effects of simulated electromagnetic fields (EMF), emitted from sub-sea power cables, on the commercially important decapod, edible crab (Cancer pagurus), were assessed. Stress related parameters were measured (l-Lactate, d-Glucose, Haemocyanin and respiration rate) along with behavioural and response parameters (antennular flicking, activity level, attraction/avoidance, shelter preference and time spent resting/roaming) during 24-h periods. Exposure to EMF had no effect on Haemocyanin concentrations, respiration rate, activity level or antennular flicking rate. EMF exposure significantly disrupted haemolymph l-Lactate and d-Glucose natural circadian rhythms. Crabs showed a clear attraction to EMF exposed shelter (69%) compared to control shelter (9%) and significantly reduced their time spent roaming by 21%. Consequently, EMF emitted from Marine Renewable Energy Devices (MREDs) will likely affect edible crabs both behaviourally and physiologically, suggesting that the impact of EMF on crustaceans must be considered when planning MREDs.


Spatial and temporal distribution of the adherens-junction-associated adhesion molecule A-CAM during avian embryogenesis.
A-CAM (adherens-junction-specific cell adhesion molecule) is a calcium-dependent adhesion molecule which is associated with intercellular adherens junctions in various tissues (Volk & Geiger, 1986, J. Cell Biol. 103, 1441-1450 and 1451-1464). In the present report, we have investigated the distribution of A-CAM during avian morphogenesis by immunofluorescence microscopy and immunoblotting. A-CAM appeared at the onset of gastrulation on developing mesodermal and endodermal cells and was then expressed on tissues derived from the three primary germ layers. During embryonic life, A-CAM was constitutively expressed in a number of tissues including the central and peripheral nervous system, myocardium, muscles, notochord, skin and lens whereas it was found transiently in many tissues ranging from the nephritic tubules and the endoderm of visceral arches to ectodermal placodes. In the adult, in addition to the nervous system, A-CAM was restricted to the skin, lens, heart and testis, and exhibited an apparent molecular weight higher than the one found in the embryo. The prevalence and cell-surface modulation of A-CAM could frequently be correlated with morphogenetic events such as mesenchyme condensation into epithelia or cell clusters (e.g. formation of the somitic epithelium, kidney tubules and peripheral ganglia), dissociation of epithelia (e.g. dissociation of the somitic epithelium and segregation of neural crest from the neural tube), separation of cell populations (e.g. fibroblasts and myotubes in the heart) and reorganizations of epithelia (e.g. neurulation). In addition, using electron microscopy, the expression of A-CAM on the surface of aggregating and separating cells could be correlated with the formation and disappearance of adherens junctions. This precisely scheduled control of A-CAM correlated with early morphogenetic events during embryogenesis suggests that this CAM could play a crucial role in these processes.


Modality-specific alpha modulations facilitate long-term memory encoding in the presence of distracters.
It has been proposed that long-term memory encoding is not only dependent on engaging task-relevant regions but also on disengaging task-irrelevant regions. In particular, oscillatory alpha activity has been shown to be involved in shaping the functional architecture of the working brain because it reflects the functional disengagement of specific regions in attention and memory tasks. We here ask if such allocation of resources by alpha oscillations generalizes to long-term memory encoding in a cross-modal setting in which we acquired the ongoing brain activity using magnetoencephalography. Participants were asked to encode pictures while ignoring simultaneously presented words and vice versa. We quantified the brain activity during rehearsal reflecting subsequent memory in the different attention conditions. The key finding was that successful long-term memory encoding is reflected by alpha power decreases in the sensory region of the to-be-attended modality and increases in the sensory region of the to-be-ignored modality to suppress distraction during rehearsal period. Our results corroborate related findings from attention studies by demonstrating that alpha activity is also important for the allocation of resources during long-term memory encoding in the presence of distracters.


Effect of long-term interferon therapy for refractory chronic hepatitis c: preventive effect on hepatocarcinogenesis.
Effect of interferon (IFN) therapy for refractory chronic hepatitis C is not sufficient. For patients with persistent hepatitis C virus (HCV) infection, one of the clinical goals is prevention of progression to liver cirrhosis (LC) and hepatocellular carcinoma (HCC). In this study, we evaluated effect of long-term IFN administration for refractory chronic hepatitis C. The patients who were positive for HCV of genotype lb in high viral load and failed in HCV elimination by standard IFN therapy were retrospectively analyzed. The patients were divided into three groups according to administration duration of IFN therapy. The patients in group 1, 2 and 3 received IFN therapy for 6 months, 6-24 months and more than 24 months, respectively. The normalization rate of alanine aminotransferase (ALT) levels less than twice that of the normal limit 6 months after the treatment was highest in group 3 (85%). The platelet counts in group 1 gradually decreased more than 3 x 10(4)/microL from the pretreatment levels at 100 months after the start of treatment. Cumulative hepatocarcinogenesis rate in groups 1, 2 and 3 were 34.7%, 5.9% and 0%, respectively. We found distinct improvement in both ALT levels and histopathological findings in the case that received the longest term of IFN therapy (91 months). Long-term IFN therapy is effective in preventing hepatocarcinogenesis through reduction of chronic necroinflammation and accumulation of fibrosis in the liver and may be a good indication even for refractory chronic hepatitis C.


Trigeminal nerve - artery contact in people without trigeminal neuralgia - MR study.
Neurovascular conflict is regarded as the most common cause of idiopathic trigeminal neuralgia. It is suspected that a blood vessel in contact with the root entry zone of a trigeminal nerve causes its irritation. The aim of the study was to evaluate how signs of neurovascular conflict can be found in people without trigeminal neuralgia The study was conducted retrospectively. The authors analyzed MR examinations of 60 patients (120 nerves), aged 22-77 years, who did not have trigeminal neuralgia or related symptoms. Nerve-artery contact at the root entry zone of a trigeminal nerve, nerve deformation, and atrophy were searched for in angio-3D-TOF images (slice thickness: 1 mm). Contact between a trigeminal nerve and an artery was found in 30 cases (25% of nerves). The blood vessel was parallel to the nerve root in 14 cases (11.7%) and crossed it at a right or acute angle in 15 cases (12.5%). Contact between artery and atrophic nerve was found in one case. Modeling of the trigeminal nerve by an artery was not found. Contact between a trigeminal nerve root and an artery in the prepontine cistern is a frequently seen anatomical variant. Therefore, detection of such a variant is not equivalent to finding the cause of a patient's complaints.


Long-term measurement of arterial blood pressure in conscious mice.
This study describes a technique for the direct daily measurement of arterial blood pressure, sampling of arterial blood, and continuous intravenous infusion in free-moving, conscious, Swiss-Webster mice. Catheters were chronically implanted in the femoral artery and vein, tunneled subcutaneously, exteriorized at the back of the neck in a lightweight tethering spring, and attached to a swivel device at the top of the cage. Time-control experiments (n = 8) demonstrated stable values of mean arterial pressure (MAP, 116 +/- 1 mmHg) and heart rate (HR, 627 +/- 21 beats/min) for up to 35 days after catheter implantation. It was further observed that restraining mice (n = 7) increased MAP by 10 +/- 3 mmHg and HR by 78 +/- 8 beats/min from the values observed under free-moving conditions. To demonstrate the chronic use of the venous catheter, intravenous infusion of NG-nitro-L-arginine methyl ester (L-NAME, 8.6 mg.kg-1.day-1, n = 6) for 5 days significantly increased MAP from 117 +/- 4 to 131 +/- 4 mmHg without altering HR. In a final group of mice (n = 5), oral L-arginine (2% in drinking water) increased plasma arginine concentration from 90 +/- 7 to 131 +/- 17 microM and prevented L-NAME hypertension. These experiments illustrate the feasibility of long-term intravenous infusion, direct arterial blood pressure measurements, and arterial blood sampling in conscious mice.


Correlates of highly active antiretroviral therapy adherence among urban Ethiopian clients.
There has been a massive expansion of highly active antiretroviral therapy (HAART) services in Ethiopia since 2005. To assess clients' self-reported adherence to HAART medication, a descriptive, comparative cross-sectional study was carried out among adults receiving HAART medication at the Zewditu Memorial Hospital ART clinic in Addis Ababa. Of 1 808 clients eligible for the study, 1 722 agreed to participate. The data were collected over six weeks in February and March 2010. Ordered and binary logistic regression models were applied to analyse the data. The majority of participants were over age 30 years, most were females, and 90% had some formal education. More than half reported being 'extremely sure' about their ability to take most or all of their medication. Self-reported adherence to the medication was generally good, as 62% said they had never missed a dose. The most commonly cited reason for missing medication was being busy (57.5%). The odds of ever missing a dose of HAART were lower for males (adjusted odds ratio [AOR]: 1.44; 95% confidence interval [CI]: 1.15-1.79), older persons (AOR: 0.98; 95% CI: 0.97-0.99), and those who did not drink alcohol regularly. Similarly, the odds of being self-confident about taking the medication properly were higher for males and for those who did not drink alcohol regularly (AOR: 0.48; 95% CI: 0.35-0.64). The odds of self-confidence in taking the medication were lower among those in lower income group. Those who reported an expenditure income of Birr 501-999 (AOR: 0.35; 95% CI: 0.24-0.49) or Birr 1 000-1 999 (AOR: 0.41; 95% CI: 0.29-0.60) had less self-confidence in taking their medication properly as compared to those who had an expenditure income of Birr 2 000 or more. There is a need to design and implement targeted adherence interventions that could lead to better treatment outcomes.


Sustained attention performance during sleep deprivation: evidence of state instability.
Nathaniel Kleitman was the first to observe that sleep deprivation in humans did not eliminate the ability to perform neurobehavioral functions, but it did make it difficult to maintain stable performance for more than a few minutes. To investigate variability in performance as a function of sleep deprivation, n = 13 subjects were tested every 2 hours on a 10-minute, sustained-attention, psychomotor vigilance task (PVT) throughout 88 hours of total sleep deprivation (TSD condition), and compared to a control group of n = 15 subjects who were permitted a 2-hour nap every 12 hours (NAP condition) throughout the 88-hour period. PVT reaction time means and standard deviations increased markedly among subjects and within each individual subject in the TSD condition relative to the NAP condition. TSD subjects also had increasingly greater performance variability as a function of time on task after 18 hours of wakefulness. During sleep deprivation, variability in PVT performance reflected a combination of normal timely responses, errors of omission (i.e., lapses), and errors of commission (i.e., responding when no stimulus was present). Errors of omission and errors of commission were highly intercorrelated across deprivation in the TSD condition (r = 0.85, p = 0.0001), suggesting that performance instability is more likely to include compensatory effort than a lack of motivation. The marked increases in PVT performance variability as sleep loss continued supports the "state instability" hypothesis, which posits that performance during sleep deprivation is increasingly variable due to the influence of sleep initiating mechanisms on the endogenous capacity to maintain attention and alertness, thereby creating an unstable state that fluctuates within seconds and that cannot be characterized as either fully awake or asleep.


Effect of relative humidity on crystal growth, device performance and hysteresis in planar heterojunction perovskite solar cells.
Due to the hygroscopic nature of organolead halide perovskites, humidity is one of the most important factors affecting the efficiency and longevity of perovskite solar cells. Although humidity has a long term detrimental effect on device performance, it also plays a key role during the initial growth of perovskite crystals. Here we demonstrate that atmospheric relative humidity (RH) plays a key role during the formation of perovskite thin films via the sequential deposition technique. Our results indicate that the RH has a substantial impact on the crystallization process, and hence on device performance. SEM and pXRD analysis show an increase in crystallite size with increasing humidity. At low RH, the formation of small cubic crystallites with large gaps between them is observed. The presence of these voids adversely affects device performance and leads to substantial hysteresis in the device. At higher RH, the perovskite crystals are larger in size, with better connectivity between the crystallites. This produced efficient planar heterojunction solar cells with low hysteresis. By careful control of the RH during the cell fabrication process, efficiencies of up to 12.2% are reached using P3HT as the hole-transport material.


Conditional Poisson models: a flexible alternative to conditional logistic case cross-over analysis.
The time stratified case cross-over approach is a popular alternative to conventional time series regression for analysing associations between time series of environmental exposures (air pollution, weather) and counts of health outcomes. These are almost always analyzed using conditional logistic regression on data expanded to case-control (case crossover) format, but this has some limitations. In particular adjusting for overdispersion and auto-correlation in the counts is not possible. It has been established that a Poisson model for counts with stratum indicators gives identical estimates to those from conditional logistic regression and does not have these limitations, but it is little used, probably because of the overheads in estimating many stratum parameters. The conditional Poisson model avoids estimating stratum parameters by conditioning on the total event count in each stratum, thus simplifying the computing and increasing the number of strata for which fitting is feasible compared with the standard unconditional Poisson model. Unlike the conditional logistic model, the conditional Poisson model does not require expanding the data, and can adjust for overdispersion and auto-correlation. It is available in Stata, R, and other packages. By applying to some real data and using simulations, we demonstrate that conditional Poisson models were simpler to code and shorter to run than are conditional logistic analyses and can be fitted to larger data sets than possible with standard Poisson models. Allowing for overdispersion or autocorrelation was possible with the conditional Poisson model but when not required this model gave identical estimates to those from conditional logistic regression. Conditional Poisson regression models provide an alternative to case crossover analysis of stratified time series data with some advantages. The conditional Poisson model can also be used in other contexts in which primary control for confounding is by fine stratification.


Triplex intermediates in folding of human telomeric quadruplexes probed by microsecond-scale molecular dynamics simulations.
We have carried out extended set of μs-scale explicit solvent MD simulations of all possible G-triplexes which can participate in folding pathways of the human telomeric quadruplex. Our study accumulates almost 60 μs of simulation data, which is by about three orders of magnitude larger sampling compared to the earlier simulations of human telomeric G-DNA triplexes. Starting structures were obtained from experimental quadruplex structures by deleting either the first or the last strand. The life-times of antiparallel triplexes with lateral and diagonal loops are at least on μs-scale, which should be sufficient to contribute to the folding pathways. However, the triplex states may involve structures with various local deviations from the ideal triplexes, such as strand tilting and various alternative and incomplete triads. The simulations reveal easy rearrangements between lateral and diagonal loop triplex topologies. Propeller loops of antiparallel triplexes may to certain extent interfere with the G-triplexes but these structures are still viable candidates to participate in the folding. In contrast, all-parallel all-anti triplexes are very unstable and are unlikely to contribute to the folding. Although our simulations demonstrate that antiparallel G-triplexes, if folded, would have life-times sufficient to participate in the quadruplex folding, the results do not rule out the possibility that the G-triplexes are out-competed by other structures not included in our study. Among them, numerous possible misfolded structures containing guanine quartets can act as off-path intermediates with longer life-times than the triplexes. Besides analyzing the structural dynamics of a diverse set of G-DNA triplexes, we also provide a brief discussion of the limitations of the simulation methodology, which is necessary for proper understanding of the simulation data.


Incorporating potency into EU classification for carcinogenicity and reproductive toxicity.
Although risk assessment, assessing the potential harm of each particular exposure of a substance, is desirable, it is not feasible in many situations. Risk assessment uses a process of hazard identification, hazard characterisation, and exposure assessment as its components. In the absence of risk assessment, the purpose of classification is to give broad guidance (through the label) on the suitability of a chemical in a range of use situations. Hazard classification in the EU is a process involving identification of the hazards of a substance, followed by comparison of those hazards (including degree of hazard) with defined criteria. Classification should therefore give guidance on degree of hazard as well as hazard identification. Potency is the most important indicator of degree of hazard and should therefore be included in classification. This is done for acute lethality and general toxicity by classifying on dose required to cause the effect. The classification in the EU for carcinogenicity and reproductive toxicity does not discriminate across the wide range of potencies seen (6 orders of magnitude) for carcinogenicity and for developmental toxicity and fertility. Therefore potency should be included in the classification process. The methodology in the EU guidelines for classification for deriving specific concentration limits is a rigorous process for assigning substances which cause tumours or developmental toxicity and infertility in experimental animals to high, medium or low degree of hazard categories by incorporating potency. Methods are suggested on how the degree of hazard so derived could be used in the EU classification process to improve hazard communication and in downstream risk management.


Synchronous intramolecular cycloadditions of the polyene macrolactam polyketide heronamide C.
A growing number of natural products appear to arise from biosynthetic pathways that involve pericyclic reactions. We show here that for the heronamides this can occur via two spontaneous pathways involving alternative thermal or photochemical intramolecular cycloadditions.


Homology Modeling and Docking Study of Shewanella-like Protein Phosphatase Involved in the Development of Ookinetes in Plasmodium.
Parasites of the genus Plasmodium cause a great deal of morbidity and mortality worldwide, largely in regions with limited access and indication to the tools necessary to control mosquito populations and to treat human infections of malaria. Five species of this class of eukaryotic pathogens cause different human diseases, with Plasmodium falciparum alone infecting approximately 500 million people per year and resulting in approximately one million deaths. The two genes encoding the Shewanella-like protein phosphatases of P. falciparum, SHLP1 and SHLP2, are conserved among members of Plasmodiidae family. SHLP is frequently found in asexual blood stages and expressed at all stages of the life cycle of parasite. SHLP deletion results in a reduction in microneme formation, ookinetes (zygote) development, and complete ablation of oocyst formation, thereby blocking transmission of parasite. Structure modeling of SHLP protein can be helpful in understanding the active site and binding site information and hence can be used for drug designing and for therapeutics against malaria. Study of SHLP and its variants was carried out using UniProtKB database. Homology modeling was performed using Schrödinger software, and the modeled structure was verified using Ramachandran plot. Ten antioxidants were searched in PubChem database for docking and comparative analysis. Docking was carried out against SHLP-modeled protein, and the ligand-protein interaction map was analyzed. Effective role of resveratrol was studied against SHLP protein using docking method to identify protein-ligand interaction scheme and bond formation. SHLP protein was modeled and docking was carried out to identify the binding sites and interaction with the SHLP protein. Docking study suggested that resveratrol has a strong interaction with SHLP protein and can be used as a potential ligand for drug designing. SHLP plays a crucial role in ookinetes and microneme development in Plasmodium; hence ligand, which can interact and inhibit SHLP protein, can be a potential drug against malarial parasite development. We studied the binding of antioxidant, such as resveratrol, with this protein-using docking method and it was found that resveratrol as an antioxidant can bind with the target SHLP protein.


Lymph Node Enlargement in Neck Filariasis as a Rare Cause: A Case Report.
Lymphatic filariasis is endemic to tropical countries and is the most common cause of acquired lymphedema in the world. Wuchereria bancrofti is the main etiological agent responsible. While the presentation of filariasis in limbs is common, isolated presentation as a single enlarged lymph node in the neck is very rare. We describe a 48-year-old Indian woman, who presented with a hard lymph node in the neck. There was no other significant lymph node enlargement. The overlying skin was erythematous, and no other findings were present on examination. Ultrasonography of the neck revealed a single enlarged lymph node in the left level 2 region, and fine-needle aspiration cytology showed microfilariae with surrounding inflammatory infiltrate. The patient was started on oral diethylcarbamazine and after 2 weeks of therapy, the lymph node enlargement in the neck subsided and the erythema was relieved.


What do consumers want to know about antibiotics? Analysis of a medicines call centre database.
Australia is one of the highest users of antibiotics in the developed world. This study aimed to identify consumer antibiotic information needs to improve targeting of medicines information. We conducted a retrospective, mixed-method study of consumers' antibiotic-related calls to Australia's National Prescribing Service (NPS) Medicines Line from September 2002 to June 2010. Demographic and question data were analysed, and the most common enquiry type in each age group was explored for key narrative themes. Relative antibiotic call frequencies were determined by comparing number of calls to antibiotic utilization in Australian Statistics on Medicines (ASM) data. Between 2002 and 2010, consumers made 8696 antibiotic calls to Medicines Line. The most common reason was questions about the role of their medicine (22.4%). Patient age groups differed in enquiry pattern, with more questions about lactation in the 0- to 4-year age group (33.6%), administration (5-14 years: 32.4%), interactions (15-24 years: 33.4% and 25-54 years: 23.3%) and role of the medicine (55 years and over: 26.6%). Key themes were identified for each age group. Relative to use in the community, antibiotics most likely to attract consumer calls were ciprofloxacin (18.0 calls/100,000 ASM prescriptions) and metronidazole (12.9 calls/100,000 ASM prescriptions), with higher call rates than the most commonly prescribed antibiotic amoxicillin (3.9 calls/100,000 ASM prescriptions). Consumers' knowledge gaps and concerns about antibiotics vary with age, and certain antibiotics generate greater concern relative to their usage. Clinicians should target medicines information to proactively address consumer concerns.


Nurse-administered procedural sedation and analgesia in the cardiac catheter laboratory: an integrative review.
To identify and appraise the literature concerning nurse-administered procedural sedation and analgesia in the cardiac catheter laboratory. An integrative review method was chosen for this study. MEDLINE and CINAHL databases as well as The Cochrane Database of Systematic Reviews and the Joanna Briggs Institute were searched. Nineteen research articles and three clinical guidelines were identified. The authors of each study reported nurse-administered sedation in the CCL is safe due to the low incidence of complications. However, a higher percentage of deeply sedated patients were reported to experience complications than moderately sedated patients. To confound this issue, one clinical guideline permits deep sedation without an anaesthetist present, while others recommend against it. All clinical guidelines recommend nurses are educated about sedation concepts. Other findings focus on pain and discomfort and the cost-savings of nurse-administered sedation, which are associated with forgoing anaesthetic services. Practice is varied due to limitations in the evidence and inconsistent clinical practice guidelines. Therefore, recommendations for research and practice have been made. Research topics include determining how and in which circumstances capnography can be used in the CCL, discerning the economic impact of sedation-related complications and developing a set of objectives for nursing education about sedation. For practice, if deep sedation is administered without an anaesthetist present, it is essential nurses are adequately trained and have access to vital equipment such as capnography to monitor ventilation because deeply sedated patients are more likely to experience complications related to sedation. These initiatives will go some way to ensuring patients receiving nurse-administered procedural sedation and analgesia for a procedure in the cardiac catheter laboratory are cared for using consistent, safe and evidence-based practices.


Promiscuity and selectivity of bitter molecules and their receptors.
Bitter taste is essential for survival, as it protects against consuming poisonous compounds, which are often bitter. Bitter taste perception is mediated by bitter taste receptors (TAS2Rs), a subfamily of G-protein coupled receptors (GPCRs). The number of TAS2R subtypes is species-dependent, and varies from 3 in chicken to 50 in frog. TAS2Rs present an intriguing case for studying promiscuity: some of the receptors are still orphan, or have few known agonists, while others can be activated by numerous, structurally dissimilar compounds. The ligands also vary in the repertoire of TAS2Rs that they activate: some bitter compounds are selective toward a single TAS2R, while others activate multiple TAS2Rs. Selectivity/promiscuity profile of bitter taste receptors and their compounds was explored by a chemoinformatic approach. TAS2R-promiscuous and TAS2R-selective bitter molecules were found to differ in chemical features, such as AlogP, E-state, total charge, number of rings, globularity, and heavy atom count. This allowed the prediction of bitter ligand selectivity toward TAS2Rs. Interestingly, while promiscuous TAS2Rs are activated by both TAS2R-promiscuous and TAS2R-selective compounds, almost all selective TAS2Rs in human are activated by promiscuous compounds, which are recognized by other TAS2Rs anyway. Thus, unique ligands, that may have been the evolutionary driving force for development of selective TAS2Rs, still need to be unraveled.


Solubility of Polar and Nonpolar Aromatic Molecules in Subcritical Water: The Role of the Dielectric Constant.
Liquid water at temperatures above the boiling point and high pressures, also known as pressurized hot water, or subcritical water (SBCW), is an effective solvent for both polar and nonpolar organic solutes. This is often associated with the decrease of water's dielectric constant at high temperatures, apparently allowing water to behave like an organic solvent. The decrease of the solubility at high pressures, in turn, is explained by a mild increase of the dielectric constant of water. Nevertheless, the relationship among the dielectric constant of water, hydration, and the solubilities of polar and nonpolar molecules in SBCW remains poorly understood. Here, we study, through molecular dynamics, the hydration thermodynamic parameters and the solubility of nonpolar and polar aromatic model systems, for which a solubility increase in SBCW is observed. We show that the temperature dependence of the hydration free energy of the model nonpolar aromatic solutes is nonmonotonic, exhibiting a solute size independent maximum at ∼475 K, above which hydration becomes entropically favorable and enthalpically unfavorable. The monotonic increase of the solubility, separated here in hydration and vaporization or sublimation components of the pure liquid or solid solute, respectively, is, in turn, related to the temperature increase of the latter, and only to a minor extent with the decrease of the hydration free energy above ∼475 K, via the hydration entropy. A solubility increase or decrease is also found at high pressures for different solutes, explained by the relative magnitude of the hydration and the vaporization or sublimation components of the solubility. For the model solid polar system studied, the hydration free energy increases monotonically with the temperature, instead, and the solubility increase is caused by the decrease of the sublimation component of the solubility. Thus, despite the observed increase of the hydration free energy with pressure, related to the entropic component decrease, our results indicate that the dielectric constant plays no significant role on the solubility increase of nonpolar and polar solutes in SBCW, opposite to the dielectric constant picture. The structure of water next to the solutes is also investigated, and a structural enhancement at room temperature is observed, resulting in significantly stronger pair interactions between a water molecule and its third and fourth nearest water neighbors. This structural and energetic enhancement nearly vanishes, however, at high temperatures, contributing to a positive hydration entropy.


Chronically Implanted Intracranial Electrodes: Tissue Reaction and Electrical Changes.
The brain-electrode interface is arguably one of the most important areas of study in neuroscience today. A stronger foundation in this topic will allow us to probe the architecture of the brain in unprecedented functional detail and augment our ability to intervene in disease states. Over many years, significant progress has been made in this field, but some obstacles have remained elusive-notably preventing glial encapsulation and electrode degradation. In this review, we discuss the tissue response to electrode implantation on acute and chronic timescales, the electrical changes that occur in electrode systems over time, and strategies that are being investigated in order to minimize the tissue response to implantation and maximize functional electrode longevity. We also highlight the current and future clinical applications and relevance of electrode technology.


STAKEHOLDER INVOLVEMENT THROUGHOUT HEALTH TECHNOLOGY ASSESSMENT: AN EXAMPLE FROM PALLIATIVE CARE.
Internationally, funders require stakeholder involvement throughout health technology assessment (HTA). We report successes, challenges, and lessons learned from extensive stakeholder involvement throughout a palliative care case study that demonstrates new concepts and methods for HTA. A 5-step "INTEGRATE-HTA Model" developed within the INTEGRATE-HTA project guided the case study. Using convenience or purposive sampling or directly / indirectly identifying and approaching individuals / groups, stakeholders participated in qualitative research or consultation meetings. During scoping, 132 stakeholders, aged ≥ 18 years in seven countries (England, Italy, Germany, The Netherlands, Norway, Lithuania, and Poland), highlighted key issues in palliative care that assisted identification of the intervention and comparator. Subsequently stakeholders in four countries participated in face-face, telephone and / or video Skype meetings to inform evidence collection and / or review assessment results. An applicability assessment to identify contextual and implementation barriers and enablers for the case study findings involved twelve professionals in the three countries. Finally, thirteen stakeholders participated in a mock decision-making meeting in England. Views about the best methods of stakeholder involvement vary internationally. Stakeholders make valuable contributions in all stages of HTA; assisting decision making about interventions, comparators, research questions; providing evidence and insights into findings, gap analyses and applicability assessments. Key challenges exist regarding inclusivity, time, and resource use. Stakeholder involvement is feasible and worthwhile throughout HTA, sometimes providing unique insights. Various methods can be used to include stakeholders, although challenges exist. Recognition of stakeholder expertise and further guidance about stakeholder consultation methods is needed.


De novo cancer in patients on dialysis and after renal transplantation: north-western Italy, 1997-2012.
Kidney transplant recipients (KTR) are known to have a higher risk of cancer than the general population, especially of malignancies related to oncogenic viral infections. This study assessed the incidence of de novo malignancies (DNMs) in patients receiving kidney transplantation and in dialysis patients (DP) on the waiting list for transplantation at the same centre. The aim was to quantify the contribution of post-transplant immunosuppression to the underlying risk of developing a DNM in dialysis patients on the waiting list for kidney transplant. Cancer incidence rates were computed using the Kaplan-Meier product-limit method. The number of DNMs observed in both groups was compared to the expected incidence in the general Italian population through calculation of the standardized incidence ratios (SIR) and their 95% confidence intervals (CI). To identify risk factors, incidence rate ratios (IRR) and 95% CIs were computed using Poisson regression analysis. The comparison of incidence rates between the two cohorts was also performed using age standardized incidence rates (ASR) and their ratio (age standardized rate ratio, ASRR). In 4858 person-years (PYs) of observation, 75 out of 735 KTR were diagnosed with DNM: 57 solid neoplasms, 13 post-transplant lymphoproliferative disorders (PTLD), and 12 Kaposi sarcomas (KS). The overall incidence was 17.5 cases/103 PYs, resulting in a 2.1-fold increased risk. Twenty-four out of 912 DP, over a follow-up of 2400 PYs, were diagnosed with a solid neoplasm, but none had PTLD or KS. The use of induction therapy after transplant was associated with a significant increased risk of KS (IRR: 3.52; 95% CI 1.04-11.98, p < 0.05). ASRR for all cancers and for solid cancers only was 1.84- and 1.19-fold higher in KTR, respectively, than in the general population. The overall incidence in DP was 10.0 cases/103 PYs, with a 1.6 significantly increased cancer risk compared to the general population. Our study confirms the increased risk of cancer after transplantation and during dialysis, but showed that virus-related cancers only occur after post-transplant immunosuppression.


Gene duplication and gene conversion in the Caenorhabditis elegans genome.
A comprehensive analysis of duplication and gene conversion for 7394 Caenorhabditis elegans genes (about half the expected total for the genome) is presented. Of the genes examined, 40% are involved in duplicated gene pairs. Intrachromosomal or cis gene duplications occur approximately two times more often than expected. In general the closer the members of duplicated gene pairs are, the more likely it is that gene orientation is conserved. Gene conversion events are detectable between only 2% of the duplicated pairs. Even given the excesses of cis duplications, there is an excess of gene conversion events between cis duplicated pairs on every chromosome except the X chromosome. The relative rates of cis and trans gene conversion and the negative correlation between conversion frequency and DNA sequence divergence for unconverted regions of converted pairs are consistent with previous experimental studies in yeast. Three recent, regional duplications, each spanning three genes are described. All three have already undergone substantial deletions spanning hundreds of base pairs. The relative rates of duplication and deletion may contribute to the compactness of the C. elegans genome.


Simple and robust image-based autofocusing for digital microscopy.
A simple image-based autofocusing scheme for digital microscopy is demonstrated that uses as few as two intermediate images to bring the sample into focus. The algorithm is adapted to a commercial inverted microscope and used to automate brightfield and fluorescence imaging of histopathology tissue sections.


Synapse organization and modulation via C1q family proteins and their receptors in the central nervous system.
Several C1q family members, related to the C1q complement component are extensively expressed in the central nervous system. Cbln1, which belongs to the Cbln subfamily of C1q proteins and released from cerebellar granule cells, plays an indispensable role in the synapse formation and function at parallel fiber-Purkinje cell synapses. This is achieved by formation of a trans-synaptic tripartite complex which is composed of one unit of the Cbln1 hexamer, monomeric neurexin (NRX) containing a splice site 4 insertion at presynaptic terminals and the postsynaptic GluD2 dimers. Recently an increasing number of soluble or transmembrane proteins have been identified to bind directly to the amino-terminal domains of iGluR and regulate the recruitment and function of iGluRs at synapses. Especially at mossy fiber (MF)-CA3 synapses in the hippocampus, postsynaptic kainate-type glutamate receptors (KARs) are involved in synaptic network activity through their characteristic channel kinetics. C1ql2 and C1ql3, which belong to the C1q-like subfamily of C1q proteins, are produced by MFs and serve as extracellular organizers to recruit functional postsynaptic KAR complexes at MF-CA3 synapses via binding to the amino-terminal domains of GluK2 and GluK4 KAR subunits. In addition, C1ql2 and C1ql3 directly bind to NRX3 containing sequences encoded by exon 25b insertion at splice site 5. In the present review, we highlighted the generality of the strategy by tripartite complex formation of the specific type of NRX and iGluR via C1q family members.


Melatonin, a natural programmed cell death inducer in cancer.
Melatonin, an indolamine derived from the amino-acid tryptophan, participates in diverse physiological functions and has great functional versatility related to the regulation of circadian rhythms and seasonal behaviour, sexual development, retinal physiology, tumour inhibition, as an antioxidant, immunomodulatory and anti-aging properties. In relation to its oncostatic properties, there is evidence that tumor initiation, promotion or progression may be restrained by the night-time physiological surge of melatonin in the blood or extracellular fluid. In addition, depressed nocturnal melatonin concentrations or nocturnal excretion of the main melatonin metabolite, 6-sulfatoxymelatonin, were found in individuals with various tumor types. In the majority of studies, melatonin was shown to inhibit development and/or growth of various experimental animal tumors and some human cell lines in vitro. Many tumors do not respond to drug treatment due to their resistance to undergo apoptosis thereby contributing to the development of cancer. Thus, given the importance of the apoptotic program in cancer treatment, the role of melatonin in influencing apoptosis in tumor cells attracted attention because it seems that it actually promotes apoptosis in most tumor cells, in contrast to the obvious inhibition of apoptotic processes in normal cells. Thus, this paper is also intended to provide to the reader an up-date of all the researches that have been carried out to date, which investigate the proapoptotic effects of melatonin in experimental preclinical models of cancer (in vitro and in vivo) and the underlying proposed action mechanism of this effects. If melatonin uniformly induces apoptosis in cancer cells, the findings could have important clinical implications to improve the quality of live while preventing the appearance of cancer.


A latent class based imputation method under Bayesian quantile regression framework using asymmetric Laplace distribution for longitudinal medication usage data with intermittent missing values.
Evaluating the association between diseases and the longitudinal pattern of pharmacological therapy has become increasingly important. However, in many longitudinal studies, self-reported medication usage data collected at patients' follow-up visits could be missing for various reasons. These pieces of missing or inaccurate/untenable information complicate determining the trajectory of medication use and its complete effects for patients. Although longitudinal models can deal with specific types of missing data, inappropriate handling of this issue can lead to a biased estimation of regression parameters especially when missing data mechanisms are complex and depend upon multiple sources of variation. We propose a latent class-based multiple imputation (MI) approach using a Bayesian quantile regression (BQR) that incorporates cluster of unobserved heterogeneity for medication usage data with intermittent missing values. Findings from our simulation study indicate that the proposed method performs better than traditional MI methods under certain scenarios of data distribution. We also demonstrate applications of the proposed method to data from the Prospective Study of Outcomes in Ankylosing Spondylitis (AS) cohort when assessing an association between longitudinal nonsteroidal anti-inflammatory drugs (NSAIDs) usage and radiographic damage in AS, while the longitudinal NSAID index data are intermittently missing.


A Multi-Cue Bioreactor to Evaluate the Inflammatory and Regenerative Capacity of Biomaterials under Flow and Stretch.
The use of resorbable biomaterials to induce regeneration directly in the body is an attractive strategy from a translational perspective. Such materials induce an inflammatory response upon implantation, which is the driver of subsequent resorption of the material and the regeneration of new tissue. This strategy, also known as in situ tissue engineering, is pursued to obtain cardiovascular replacements such as tissue-engineered vascular grafts. Both the inflammatory and the regenerative processes are determined by the local biomechanical cues on the scaffold (i.e., stretch and shear stress). Here, we describe in detail the use of a custom-developed bioreactor that uniquely enables the decoupling of stretch and shear stress on a tubular scaffold. This allows for the systematic and standardized evaluation of the inflammatory and regenerative capacity of tubular scaffolds under the influence of well-controlled mechanical loads, which we demonstrate on the basis of a dynamic co-culture experiment using human macrophages and myofibroblasts. The key practical steps in this approach-the construction and setting up of the bioreactor, preparation of the scaffolds and cell seeding, application and maintenance of stretch and shear flow, and sample harvesting for analysis-are discussed in detail.


Does the clock drawing test predict dementia? Results of the Leipzig longitudinal study of the aged (LEILA 75+).
Conversion rates to dementia are known to be high for patients with mild cognitive impairment (MCI), but the diagnosis of MCI is very time-consuming. Since the Clock Drawing Test (CDT) is quick to administer, it will be of interest to compare the predictive validity of the CDT and of an MCI diagnosis for the diagnosis of dementia. In a sample of 384 individuals, CDT scores and the presence of MCI were assessed at baseline and then compared between individuals with an incident dementia diagnosis at follow-up and those without. Multivariate analyses, receiver operating characteristic analyses and values of sensitivity and specificity of the CDT were performed. Individuals with incident dementia had significantly higher CDT scores at baseline than those without dementia. CDT was a significant predictor of incident dementia after adjusting for other factors. CDT reached a sensitivity of 68% and specificity of 65%. The area under the receiver operating characteristic curve of CDT was 0.70 and therefore slightly lower than for MCI diagnosis (0.78). Because of the only slightly lower predictive value of the CDT, its quick application and scoring compared to the MCI concept applied, it will be worthwhile to improve the CDT scoring system in order to increase the predictive validity in dementia.


Welfare of pigs during killing for purposes other than slaughter.
Pigs at different stages of the production cycle may have to be killed on-farm for purposes other than slaughter (where slaughter is defined as killing for human consumption) either individually (e.g. severely injured pigs) or on a large scale (e.g. unproductive animals or for disease control reasons). This opinion assessed the risks associated with the on-farm killing of pigs and included two phases: 1) handling and moving of pigs and 2) killing methods (including restraint). The killing methods were subdivided into four categories: electrical methods, mechanical methods, gas mixture methods and lethal injection. Four welfare consequences to which pigs can be exposed to during on-farm killing were identified: pain, fear, impeded movement and respiratory distress. Welfare consequences and relevant animal-based measures were described. In total, 28 hazards were associated with the welfare consequences; majority of the hazards (24) are related to Phase 2 (killing). The main hazards are associated with lack of staff skills and training, and poor-designed and constructed facilities. Staff was identified as an origin of all hazards, either due to lack of skills needed to perform appropriate killing or due to fatigue. Corrective measures were identified for 25 hazards. Outcome tables linking hazards, welfare consequences, animal-based measures, hazard origins, preventive and corrective measures were developed and mitigation measures proposed.


Use of supramaximal stimulation to predict facial nerve outcomes following vestibular schwannoma microsurgery: results from a decade of experience.
The goal of vestibular schwannoma surgery is tumor removal and preservation of neural function. Intraoperative facial nerve (FN) monitoring has emerged as the standard of care, but its role in predicting long-term facial function remains a matter of debate. The present report seeks to describe and critically assess the value of applying current at supramaximal levels in an effort to identify patients destined for permanent facial paralysis. Over more than a decade, the protocol for stimulating and assessing the FN during vestibular schwannoma surgery at the authors' institution has consisted of applying pulsed constant-current stimulation at supramaximal levels proximally and distally following tumor resection to generate an amplitude ratio, which subtracted from 100% yields the degree to which the functional integrity of the FN "dropped off" intraoperatively. These data were prospectively collected and additional variables that might impact postoperative FN function were retrospectively reviewed from the medical record. Only patients with anatomically intact FNs and > 12 months of follow-up data were analyzed. There were 267 patients available for review. The average posterior fossa tumor diameter was 24 mm and the rate of long-term good (House-Brackmann Grade I-II) FN function was 84%. Univariate logistic regression analysis revealed that prior treatment, neurofibromatosis Type 2 status, tumor size, cerebellopontine angle extension, subjectively thinned FN at the time of operation, minimal stimulation threshold, percent dropoff by supramaximal stimulation (SMS), and postoperative FN function all correlated statistically (p < 0.05) with long-term FN function. When evaluating patients with significant FN weakness at the time of hospital discharge, only the percent dropoff by SMS remained a significant predictor of long-term FN function. However, the positive predictive value of SMS for long-term weakness is low, at 46%. In a large cohort of patients, the authors found that interrogating intraoperative FN function with SMS is safe and technically simple. It is useful for predicting which patients will ultimately have good facial function, but is very limited in identifying patients destined for long-term facial weakness. This test may prove helpful in the future in tailoring less than gross-total tumor removal to limit postoperative facial weakness.


Association of Intracranial Atherosclerotic Disease With Brain β-Amyloid Deposition: Secondary Analysis of the ARIC Study.
Intracranial atherosclerotic disease (ICAD) is an important cause of stroke and has also been recently identified as an important risk factor for all-cause dementia, but the mechanism of its association with cognitive performance is not fully understood. To test the hypothesis that ICAD is associated with cerebral β-amyloid deposition as a marker of Alzheimer disease. This cross-sectional analysis of data collected from August 2011 through November 2014 was a community-based cohort study conducted in 3 US communities. Of 346 adults without dementia aged 70 to 90 years who were sequentially recruited from 3 of 4 sites of the larger Atherosclerosis Risk in Communities study into a study of brain florbetapir positron emission tomography (ARIC-PET), 300 met inclusion criteria. A total of 589 were approached about recruitment, of whom 346 (58.7%) consented (the remainder either met exclusion criteria for ARIC-PET or refused to participate). Data were analyzed from July 2017 through October 2019. Intracranial atherosclerotic disease presence, frequency, and extent of stenosis, by high-resolution vessel wall magnetic resonance imaging. Global cortical standardized uptake value ratio (SUVR) of greater than 1.2 as measured by florbetapir PET. Models were conducted using logistic regression methods. In secondary analyses, we tested effect modifications by apolipoprotein E ε4 genotype with interaction terms and in stratified models and evaluated regional patterns of associations. In 300 participants (mean [SD] age, 76 [5] years; 132 African American individuals [44%], 167 women [56%], and 94 carriers of at least 1 apolipoprotein E ε4 allele [31%]), ICAD was found in 105 participants (35%) and mean (SD) SUVR was higher in individuals with vs without intracranial plaques (1.34 [0.29] vs 1.27 [0.23]; P = .03). In adjusted models, ICAD presence (plaque presence [adjusted odds ratio (aOR), 1.20; 95% CI, 0.69-2.07] and frequency [aOR, 1.10; 95% CI, 0.96-1.26]) was not associated significantly with elevated SUVR in the total sample. Furthermore, modest stenosis of the intracranial vessels (defined as >50% stenosis) was not associated with elevated SUVR (aOR, 2.33; 95% CI, 0.82-6.60). In this community-based cohort of adults without dementia, intracranial atherosclerotic plaque or stenosis was not associated with brain β-amyloid deposition.


Challenges in developing paediatric surgery in Sub-Saharan Africa.
The aim of the present study is to report on the difficulties in developing paediatric surgery in a teaching hospital in Tanzania. The methods are as follows: (1) information on the demography and health services of Tanzania were obtained from the Ministry of Health and the 1998 consensus report; (2) hospital data was obtained from the Medical Director's Office and analysed; (3) the current delivery of surgical services for children at the Tumaini University Hospital and Kilimanjaro Christian Medical Centre (KCMC) is reported; (4) the local, national and international support for the development of surgical services for children is noted; (5) the teaching, training and research programmes are proposed. The results showed that (1) Tanzania has a population of 31 million with a total health expenditure of 10%. There is 1 doctor for 23,000 inhabitants and 1 hospital bed per 940. The infant mortality is 173 per thousand life births. (2) The bed capacity at KCMC is 500 with a staff of 40 consultants, 294 nurses, 246 health attendants and 38 clinical officers. (3) Forty-two percent of admissions to the surgical ward and 50% of surgical outpatients are children. Surgical newborns are cared for in the special care baby unit and there are no neonatal or paediatric ventilators. (4) Support to develop surgical services for children has been pledged for locally, nationally and internationally; however, delivery of the service has not had the equivalent momentum. (5) The proposed teaching and training programme has been approved by the University. There is a need for surgical services for children with encouraging support for this venture; however, the challenges remain in delivering the service.


Genetically Encoded Inverse Bolaamphiphiles.
Developing new protein-based materials with a programmable assembly is of great scientific interest and technological importance. Inspired by nature's use of post-translational modifications (PTMs) to control the function and location of proteins, we have leveraged lipidation-the PTM of proteins with lipids-to synthesize genetically encoded lipidated proteins with controllable hierarchical assembly. Specifically, we envisioned the combination of two orthogonal lipidation pathways with different regioselectivity and substrate preferences inside Escherichia coli to produce recombinant nanomaterials with distinct lipidation domains at each terminus of proteins. In this study, we demonstrate the orthogonality of N-myristoylation and C-cholesterylation pathways for recombinant production of lipidated proteins with a unique triblock architecture, which is a hydrophilic protein block flanked by two lipid tails, i.e., inverse bolaamphiphiles. Our study indicates that the architecture of lipidated protein and the sequence of the polypeptide can be used to control the hierarchical self-assembly of these materials. We envision this bio-enabled approach yielding unexplored recombinant hybrid biomaterials with tunable nanoscale structure and morphology with applications in nanobiotechnology.


A versatile methodology for the controlled synthesis of photoluminescent high-boron-content dendrimers.
Fluorescent star-shaped molecules and dendrimers with a 1,3,5-triphenylbenzene moiety as the core and 3 or 9 carborane derivatives at the periphery, have been prepared in very good yields by following different approaches. One procedure relies on the nucleophilic substitution of Br groups in 1,3,5-tris(4-(3-bromopropoxy)phenyl)benzene with the monolithium salts of methyl and phenyl-o-carborane. The second method is the hydrosilylation reactions on the peripheral allyl ether functions of 1,3,5-tris(4-allyloxy-phenyl)benzene and 1,3,5-tris(4-(3,4,5-trisallyloxybenzyloxy)phenyl)benzene with suitable carboranyl-silanes to produce different generations of dendrimers decorated with carboranyl fragments. This approach is very versatile and allows one to introduce long spacers between the fluorescent cores and the boron clusters, as well as to obtain a high loading of boron clusters. The removal of one boron atom from each cluster leads to high-boron-content water-soluble macromolecules. Thermogravimetric analyses show a higher thermal stability for the three-functionalized compounds than for those containing 9 clusters. All compounds exhibit photoluminescent properties at room temperature under ultraviolet irradiation with high quantum yields; these depend on the nature of the cluster and the substituent on the C(cluster). Cyclic voltammetry indicates that there is no electronic communication between the core and the peripheral carboranyl fragments. Due to the high boron content of these molecules, we currently focus our research on their biocompatibility, biodistribution in cells cultures, and potential applications for boron neutron capture therapy (BNCT).


Next generation Advanced Laser Fluorometry (ALF) for characterization of natural aquatic environments: new instruments.
The new optical design allows single- or multi-wavelength excitation of laser-stimulated emission (LSE), provides optimized LSE optical collection for spectral and temporal analyses, and incorporates swappable modules for flow-through and small-volume sample measurements. The basic instrument configuration uses 510 nm laser excitation for assessments of chlorophyll-a, phycobiliprotein pigments, variable fluorescence (F(v)/F(m)) and chromophoric dissolved organic matter (CDOM) in CDOM-rich waters. The three-laser instrument configuration (375, 405, and 510 nm excitation) provides additional Fv/Fm measurements with 405 nm excitation, CDOM assessments in a broad concentration range, and potential for spectral discrimination between oil and CDOM fluorescence. The new measurement protocols, analytical algorithms and examples of laboratory and field measurements are discussed.


The management of invasive bladder cancer: a review of selected Australasian studies in radiotherapy, chemotherapy and cystectomy.
Since 1968, studies have been conducted in Australia and New Zealand to improve the results of treatment for invasive bladder cancer. The prognostic value of clinical stage and of ureteric obstruction has been confirmed. Radiotherapy has been established as appropriate treatment for the majority of patients with bladder cancer, most of whom are elderly and suffer from other health problems. There has been no additional benefit from radiosensitizers in increasing the effectiveness of radiotherapy such as hyperbaric oxygen or misonidazole, or from irradiation of the para-aortic nodes, all of which can result in additional toxicity without increasing tumour control. Pre-emptive chemotherapy with cisplatin has had no obvious impact on improving survival. Elective total cystectomy has shown the prognostic value of tumour downstaging by pre-operative pelvic radiotherapy. Although younger patients are usually selected for total cystectomy and urinary diversion, there is no evidence that, stage for stage, elective total cystectomy confers a survival advantage compared with radical pelvic radiotherapy.


Hippo Enters the Competition.
In this issue of Developmental Cell, Hashimoto and Sasaki (2019) explore the role of the Hippo pathway in the establishment of naive pluripotency and cell competition in the epiblast. Their work gives insight into how the mouse embryo selects cells with the highest future potential, through the activity of YAP/TEAD.


Functional analysis of Wolbachia Cid effectors unravels cooperative interactions to target host chromatin during replication.
Wolbachia are common bacteria among terrestrial arthropods. These endosymbionts transmitted through the female germline manipulate their host reproduction through several mechanisms whose most prevalent form called Cytoplasmic Incompatibility -CI- is a conditional sterility syndrome eventually favoring the infected progeny. Upon fertilization, the sperm derived from an infected male is only compatible with an egg harboring a compatible Wolbachia strain, this sperm leading otherwise to embryonic death. The Wolbachia Cif factors CidA and CidB responsible for CI and its neutralization function as a Toxin-Antitoxin system in the mosquito host Culex pipiens. However, the mechanism of CidB toxicity and its neutralization by the CidA antitoxin remain unexplored. Using transfected insect cell lines to perform a structure-function analysis of these effectors, we show that both CidA and CidB are chromatin interactors and CidA anchors CidB to the chromatin in a cell-cycle dependent-manner. In absence of CidA, the CidB toxin localizes to its own chromatin microenvironment and acts by preventing S-phase completion, independently of its deubiquitylase -DUB- domain. Experiments with transgenic Drosophila show that CidB DUB domain is required together with CidA during spermatogenesis to stabilize the CidA-CidB complex. Our study defines CidB functional regions and paves the way to elucidate the mechanism of its toxicity.


Association of the autoimmune disease scleroderma with an immunologic response to cancer.
Autoimmune diseases are thought to be initiated by exposures to foreign antigens that cross-react with endogenous molecules. Scleroderma is an autoimmune connective tissue disease in which patients make antibodies to a limited group of autoantigens, including RPC1, encoded by the POLR3A gene. As patients with scleroderma and antibodies against RPC1 are at increased risk for cancer, we hypothesized that the "foreign" antigens in this autoimmune disease are encoded by somatically mutated genes in the patients' incipient cancers. Studying cancers from scleroderma patients, we found genetic alterations of the POLR3A locus in six of eight patients with antibodies to RPC1 but not in eight patients without antibodies to RPC1. Analyses of peripheral blood lymphocytes and serum suggested that POLR3A mutations triggered cellular immunity and cross-reactive humoral immune responses. These results offer insight into the pathogenesis of scleroderma and provide support for the idea that acquired immunity helps to control naturally occurring cancers.


Not just a circle: flux modes in the plant TCA cycle.
The tricarboxylic acid (TCA) cycle is one of the iconic pathways in metabolism. The cycle is commonly thought of in terms of energy metabolism, being responsible for the oxidation of respiratory substrates to drive ATP synthesis. However, the reactions of carboxylic acid metabolism are embedded in a larger metabolic network and the conventional TCA cycle is only one way in which the component reactions can be organised. Recent evidence from labelling studies and metabolic network models suggest that the organisation of carboxylic acid metabolism in plants is highly dependent on the metabolic and physiological demands of the cell. Thus, alternative, non-cyclic flux modes occur in leaves in the light, in some developing oilseeds, and under specific physiological circumstances such as anoxia.


Ultrastructural differentiation of sperm tail region in Diplometopon zarudnyi (an amphisbaenian reptile).
Diplometopon zarudnyi, a worm lizard belongs to amphisbaenia under trogonophidae family. This species exists in limited areas of the Arabian Peninsula and is an oscillating digger found in sub-surface soils. The present study aimed to investigate the sperm tail differentiation in D. zarudnyi. Ten male adults of D. zarudnyi were collected from Riyadh during April-May 2011. To study the sperm tail at the ultrastructural level the testes were fixed in 3% glutaraldehyde, than post fixed in 1% osmium tetaroxide followed by dehydration in ethanol grades; samples were cleared in propylene oxide and embedded in resin. Tail formation begins by the moving of centrioles and mitochondria towards the posterior pole of sperm head. Simultaneously many microtubules of the midpiece axoneme were enclosed by a thick layer of granular material. Mitochondria of midpiece lie alongside the proximal centriole which forms a very short neck region and possess tubular cristae internally and concentric layers of cristae superficially. During this course a fibrous sheath surrounds the axoneme of mid and principal piece. At the end dissolution of longitudinal manchette takes place. The mitochondria then rearrange themselves around the proximal and distal centrioles to form a neck region. Later, the fibrous sheath surrounds the proximal portion of the flagella. This part along with sperm head of D. zarudnyi provides a classical model that could be used in future for evolutionary and phylogenetic purposes of class reptilia.


Field-testing of guidance on the appropriate labelling of processed complementary foods for infants and young children in South Africa.
There is a lack of formal guidance from international normative bodies on the appropriate marketing of processed complementary foods. Such guidance is necessary to protect and promote optimal infant and young child feeding practices. The aim of this study was to field-test, in South Africa, the interim guidance provided by the Maternal, Infant and Young Child Nutrition Working Group's Draft Guide for Marketing Complementary Foods as a potential tool for use by manufacturers and national governments for guiding the appropriate labelling (as a subset of appropriate marketing practices) of complementary foods. This guidance was used to develop a checklist of questions and criteria for each possible answer, which was tested using a comprehensive database of labels from products purchased in South Africa from June to August 2011. One hundred and sixty product labels of 35 manufacturers were analysed, none of which complied with all checklist criteria. Fifty-six (35%) labels did not provide an appropriate age of introduction while 37 (23%) used images of infants appearing younger than 6 months. Nineteen (12%) labels suggested a daily ration too large for a breastfed child, and 32 (20%) potentially promote the manufacturer's infant formula. Only 58 (36%) labels were easy to read. The majority (69% and 92%) of labels provided instructions for safe and appropriate preparation/use and storage, respectively. The Draft Guide for Marketing Complementary Foods was found to be a useful tool for guiding the appropriate labelling of complementary foods, although some changes and additions are recommended to improve understanding, ease of use and to minimise the subjective interpretation of the guidance.


Multimodal imaging and photothermal therapy were simultaneously achieved in the core-shell UCNR structure by using single near-infrared light.
Core-shell nanostructures consisting of plasmonic materials and lanthanide-doped upconversion nanoparticles (UCNPs) show promising applications in theranostics including bio-imaging, diagnosis and therapy. However, some challenges still remain in the synthetic control because of the non-coordination between energy transfer and photothermal therapy (PTT). Herein, we developed a novel type of thermal-fluorescent core-shell hybrid nanocomposite incorporating rare-earth Yb3+ and Er3+ ion doped GdOF as the shell and gold nanorods (GNRs) as the core, creating upconversion nanorods (UCNRs) of GNRs@GdOF:Yb3+,Er3+. In order to facilitate the absorption or excretion of UCNRs in vivo, we designed gold nanorods with lower aspect ratios by reducing the amount of CTAB in the growth solution. More importantly, under 980 nm near-infrared (NIR) light irradiation, the green and red emissions of GdOF:Yb3+,Er3+ generally overlap with the visible absorbance of GNRs; by altering the contents of Yb3+ and Er3+ ions appropriately, the localized surface plasmon resonance (LSPR) absorption of low aspect ratio GNRs under 980 nm NIR laser excitation can be enhanced for improving the PTT efficiency. Furthermore, in vitro and in vivo assays reveal that the composite has excellent bio-compatibility and cancer therapy efficiency. This multi-functional nanocomposite, which possesses upconversion luminescence and photothermal and biocompatibility properties, shows strong potential for application in bio-imaging and photothermal anti-cancer therapy.


Dectin-2 Is a C-Type Lectin Receptor that Recognizes Pneumocystis and Participates in Innate Immune Responses.
Pneumocystis is an important fungal pathogen that causes life-threatening pneumonia in patients with AIDS and malignancy. Lung fungal pathogens are recognized by C-type lectin receptors (CLRs), which bind specific ligands and stimulate innate immune responses. The CLR Dectin-1 was previously shown to mediate immune responses to Pneumocystis spp. For this reason, we investigated a potential role for Dectin-2. Rats with Pneumocystis pneumonia (PCP) exhibited elevated Dectin-2 mRNA levels. Soluble Dectin-2 carbohydrate-recognition domain fusion protein showed binding to intact Pneumocystis carinii (Pc) and to native Pneumocystis major surface glycoprotein/glycoprotein A (Msg/gpA). RAW macrophage cells expressing V5-tagged Dectin-2 displayed enhanced binding to Pc and increased protein tyrosine phosphorylation. Furthermore, the binding of Pc to Dectin-2 resulted in Fc receptor-γ-mediated intracellular signaling. Alveolar macrophages from Dectin-2-deficient mice (Dectin-2-/-) showed significant decreases in phospho-Syk activation after challenge with Pc cell wall components. Stimulation of Dectin-2-/- alveolar macrophages with Pc components showed significant decreases in the proinflammatory cytokines IL-6 and TNF-α. Finally, during infection with Pneumocystis murina, Dectin-2-/- mice displayed downregulated mRNA expression profiles of other CLRs implicated in fungal immunity. Although Dectin-2-/- alveolar macrophages had reduced proinflammatory cytokine release in vitro, Dectin-2-/- deficiency did not reduce the overall resistance of these mice in the PCP model, and organism burdens were statistically similar in the long-term immunocompromised and short-term immunocompetent PCP models. These results suggest that Dectin-2 participates in the initial innate immune signaling response to Pneumocystis, but its deficiency does not impair resistance to the organism.


LRIG1 is a triple threat: ERBB negative regulator, intestinal stem cell marker and tumour suppressor.
In baseball parlance, a triple threat is a person who can run, hit and throw with aplomb. Leucine-rich repeats and immunoglobulin-like domains 1 (LRIG1) is a cell surface protein that antagonises ERBB receptor signalling by downregulating receptor levels. Over 10 years ago, Hedman et al postulated that LRIG1 might be a tumour suppressor. Recently, Powell et al provided in vivo evidence substantiating that claim by demonstrating that Lrig1 loss in mice leads to spontaneously arising, highly penetrant intestinal adenomas. Interestingly, Lrig1 also marks stem cells in the gut, suggesting a potential role for Lrig1 in maintaining intestinal epithelial homeostasis. In this review, we will discuss the ability of LRIG1 to act as a triple threat: pan-ERBB negative regulator, intestinal stem cell marker and tumour suppressor. We will summarise studies of LRIG1 expression in human cancers and discuss possible related roles for LRIG2 and LRIG3.


Antiviral vaccines license T cell responses by suppressing granzyme B levels in human plasmacytoid dendritic cells.
Human plasmacytoid dendritic cells (pDC) are important modulators of adaptive T cell responses during viral infections. Recently, we found that human pDC produce the serine protease granzyme B (GrB), thereby regulating T cell proliferation in a GrB-dependent manner. In this study, we demonstrate that intrinsic GrB production by pDC is significantly inhibited in vitro and in vivo by clinically used vaccines against viral infections such as tick-borne encephalitis. We show that pDC GrB levels inversely correlate with the proliferative response of coincubated T cells and that GrB suppression by a specific Ab or a GrB substrate inhibitor results in enhanced T cell proliferation, suggesting a predominant role of GrB in pDC-dependent T cell licensing. Functionally, we demonstrate that GrB(high) but not GrB(low) pDC transfer GrB to T cells and may degrade the ζ-chain of the TCR in a GrB-dependent fashion, thereby providing a possible explanation for the observed T cell suppression by GrB-expressing pDC. Modulation of pDC-derived GrB activity represents a previously unknown mechanism by which both antiviral and vaccine-induced T cell responses may be regulated in vivo. Our results provide novel insights into pDC biology during vaccinations and may contribute to an improvement of prophylactic and therapeutic vaccines.


Effect of dietary fat on endocannabinoids and related mediators: consequences on energy homeostasis, inflammation and mood.
Among the several known fatty acid-derived chemical signals, the endogenous ligands of cannabinoid receptors type-1 and -2, two G-protein-coupled receptors involved in several aspects of mammalian physiology and pathology, are perhaps those the levels of which have proven to be most sensitive to the fatty acid composition of the diet. The two most studied such ligands, known as endocannabinoids, are N-arachidonoyl-ethanolamine and 2-archidonoylglycerol, and are found in tissues together with other N-acyl-ethanolamines and 2-acylglycerols, not all of which activate the cannabinoid receptors, although several of them do exhibit important pharmacological effects. In this review article, we describe literature data indicating that the tissue concentrations of the endocannabinoids and related signalling molecules, and hence the activity of the respective receptors, can be modulated by modifying the fatty acid composition of the diet, and particularly its content in long chain PUFAs or in long chain PUFA precursors. We also discuss the potential impact of these diet-induced changes of endocannabinoid tone on three of the major pathological conditions in which cannabinoid receptors have been involved, that is metabolic dysfunctions, inflammation and affective disorders.


Clinical and hematologic manifestations in patients with Diamond Blackfan anemia in Korea.
Diamond Blackfan anemia (DBA), characterized by impaired red cell production, is a rare condition that is usually symptomatic in early infancy. The purpose of this study was to assess nationwide experiences of DBA encountered over a period of 20 years. The medical records of 56 patients diagnosed with DBA were retrospectively reviewed from November 1984 to July 2010. Fifteen institutions, including 13 university hospitals, participated in this study. The male-to-female ratio of patients with DBA was 1.67:1. The median age of diagnosis was 4 months, and 74.1% were diagnosed before 1 year of age. From 2000 to 2009, annual incidence was 6.6 cases per million. Excluding growth retardation, 38.2% showed congenital defects: thumb deformities, ptosis, coarctation of aorta, ventricular septal defect, strabismus, etc. The mean hemoglobin concentration was 5.1±1.9 g/dL, mean corpuscular volume was 93.4±11.6 fL, and mean number of reticulocytes was 19,700/mm(3). The mean cellularity of bone marrow was 75%, with myeloid:erythroid ratio of 20.4:1. After remission, 48.9% of patients did not need further steroids. Five patients with DBA who received hematopoietic transplantation have survived. Cancer developed in 2 cases (3.6%). The incidence of DBA is similar to data already published, but our study had a male predilection. Although all patients responded to initial treatment with steroids, about half needed further steroids after remission. It is necessary to collect further data, including information regarding management pathways, from nationwide DBA registries, along with data on molecular analyses.


Functional polymorphisms associated with disease-free survival in resected carcinoma of the esophagus.
The aim of this study was to determine whether clinical outcome after surgical resection of esophageal adenocarcinoma (EAC) or esophageal squamous cell carcinoma (ESCC) could be predicted by functional polymorphisms in different proto-oncogenes and tumor suppressor genes. Six single nucleotide polymorphisms (SNPs) in the AURKA (rs2273535), ERBB2 (rs1136201), MDM2 (rs2279744), CDH1 (rs5030625), CDKN2A (rs11515), and TP73 (rs2273953) genes were genotyped in a consecutive cohort of 346 esophageal cancer patients, who had underwent surgical resection with curative intent. Associations with disease-free survival (DFS) were analyzed with Kaplan-Meier curves and Cox regression, adjusting for potential confounders. Univariate analysis showed no significant associations between the tested polymorphisms and DFS in patients with EAC or ESCC. However, in a multivariate analysis, patients with EAC carrying the heterozygous MDM2 (rs2279744) T/G genotype had significantly improved DFS compared with patients carrying the wild-type genotype (adjusted hazard ratio (AHR), 0.63; 95% confidence interval (CI) [0.45-0.88]). Patients with EAC harboring the homozygous CDH1 (rs5030625) GA/GA genotype had a significantly reduced survival as compared with patients carrying the wild-type genotype AHR 4.0, 95% CI [1.4-11]. In a large cohort of esophageal cancer patients, the MDM2 T/G and CDH1 GA/GA genotype confer risk of death in patients with EAC. These data suggest that inter-individual differences in germ-line DNA have an impact on DFS in patients with EAC.


Structural and motional contributions of the Bacillus subtilis ClpC N-domain to adaptor protein interactions.
The AAA(+) (ATPases associated with a variety of cellular activities) superfamily protein ClpC is a key regulator of cell development in Bacillus subtilis. As part of a large oligomeric complex, ClpC controls an array of cellular processes by recognizing, unfolding, and providing misfolded and aggregated proteins as substrates for the ClpP peptidase. ClpC is unique compared to other HSP100/Clp proteins, as it requires an adaptor protein for all fundamental activities. The NMR solution structure of the N-terminal repeat domain of ClpC (N-ClpCR) comprises two structural repeats of a four-helix motif. NMR experiments used to map the MecA adaptor protein interaction surface of N-ClpCR reveal that regions involved in the interaction possess conformational flexibility and conformational exchange on the microsecond-to-millisecond timescale. The electrostatic surface of N-ClpCR differs substantially from the N-domain of Escherichia coli ClpA and ClpB, suggesting that the electrostatic surface characteristics of HSP100/Clp N-domains may play a role in adaptor protein and substrate interaction specificity, and perhaps contribute to the unique adaptor protein requirement of ClpC.


Trisomic pregnancy and earlier age at menopause.
We tested the hypothesis that the connection between advanced maternal age and autosomal trisomy reflects the diminution of the oocyte pool with age. Because menopause occurs when the number of oocytes falls below some threshold, our hypothesis is that menopause occurs at an earlier age among women with trisomic pregnancies than it does among women with chromosomally normal pregnancies. To determine their menstrual status, we interviewed women from our previous study of karyotyped spontaneous abortions who, in 1993, were age >/=44 years. Premenopausal women completed interviews every 4-5 mo, until menopause or until the study ended in 1997. The primary analyses compare 111 women whose index pregnancy was a trisomic spontaneous abortion with two groups: women whose index pregnancy was a chromosomally normal loss (n=157) and women whose index pregnancy was a chromosomally normal birth (n=226). We used a parametric logistic survival analysis to compare median ages at menopause. The estimated median age at menopause was 0.96 years earlier (95% confidence interval -0.18 to 2.10) among women with trisomic losses than it was among women with chromosomally normal losses and chromosomally normal births combined. Results were unaltered by adjustment for education, ethnicity, and cigarette smoking. Our results support the hypothesis that trisomy risk is increased with decreased numbers of oocytes. Decreased numbers may indicate accelerated oocyte atresia or fewer oocytes formed during fetal development.


Alterations in ubiquitin ligase Siah-2 and its corepressor N-CoR after P-MAPA immunotherapy and anti-androgen therapy: new therapeutic opportunities for non-muscle invasive bladder cancer.
The present study describes the role of the ubiquitin ligase Siah-2 and corepressor N-CoR in controlling androgen receptor (AR) and estrogen receptors (ERα and ERβ) signaling in an appropriate animal model (Fischer 344 female rats) of non-muscle invasive bladder cancer (NMIBC), especially under conditions of anti-androgen therapy with flutamide. Furthermore, this study describes the mechanisms of a promising therapeutic alternative for NMIBC based on Protein aggregate magnesium-ammonium phospholinoleate-palmitoleate anhydride (P-MAPA) intravesical immunotherapy combined with flutamide, involving the interaction among steroid hormone receptors, their regulators and Toll-like receptors (TLRs). Our results demonstrated that increased Siah-2 and AR protein levels and decreased N-CoR, cytochrome P450 (CYP450) and estrogen receptors levels played a critical role in the urothelial carcinogenesis, probably leading to escape of urothelial cancer cells from immune system attack. P-MAPA immunotherapy led to distinct activation of innate immune system TLRs 2 and 4-mediated, resulting in increase of interferon signaling pathway, which was more effective in recovering the immunosuppressive tumor immune microenvironment and in recovering the bladder histology features than BCG (Bacillus Calmette-Guerin) treatments. The AR blockade therapy was important in the modulating of downstream molecules of TLR2 and TLR4 signaling pathway, decreasing the inflammatory cytokines signaling and enhancing the interferon signaling pathway when associated with P-MAPA. Taken together, the data obtained suggest that interferon signaling pathway activation and targeting AR and Siah-2 signals by P-MAPA intravesical immunotherapy alone and/ or in combination with AR blockade may provide novel therapeutic approaches for NMIBC.