Proteasome activity is required for the stage-specific transformation of a protozoan parasite.
A prominent feature of the life cycle of intracellular parasites is the profound morphological changes they undergo during development in the vertebrate and invertebrate hosts. In eukaryotic cells, most cytoplasmic proteins are degraded in proteasomes. Here, we show that the transformation in axenic medium of trypomastigotes of Trypanosoma cruzi into amastigote-like organisms, and the intracellular development of the parasite from amastigotes into trypomastigotes, are prevented by lactacystin, or by a peptide aldehyde that inhibits proteasome function. Clasto-lactacystin, an inactive analogue of lactacystin, and cell-permeant peptide aldehyde inhibitors of T. cruzi cysteine proteinases have no effect. We have also identified the 20S proteasomes from T. cruzi as a target of lactacystin in vivo. Our results document the essential role of proteasomes in the stage-specific transformation of a protozoan.


A Potential Role for α-Amylase in Amyloid-β-Induced Astrocytic Glycogenolysis and Activation.
Astrocytes produce and store the energy reserve glycogen. However, abnormal large glycogen units accumulate if the production or degradation of glycogen is disturbed, a finding often seen in patients with Alzheimer's disease (AD). We have shown increased activity of glycogen degrading α-amylase in AD patients and α-amylase positive glial cells adjacent to AD characteristic amyloid-β (Aβ) plaques. Investigate the role of α-amylase in astrocytic glycogenolysis in presence of Aβ. Presence of α-amylase and large glycogen units in postmortem entorhinal cortex from AD patients and non-demented controls were analyzed by immunohistological stainings. Impact of different Aβ42 aggregation forms on enzymatic activity (α-amylase, pyruvate kinase, and lactate dehydrogenase), lactate secretion, and accumulation of large glycogen units in cultured astrocytes were analyzed by activity assays, ELISA, and immunocytochemistry, respectively. AD patients showed increased number of α-amylase positive glial cells. The glial cells co-expressed the astrocytic marker glial fibrillary acidic protein, displayed hypertrophic features, and increased amount of large glycogen units. We further found increased load of large glycogen units, α-amylase immunoreactivity and α-amylase activity in cultured astrocytes stimulated with fibril Aβ42, with increased pyruvate kinase activity, but unaltered lactate release as downstream events. The fibril Aβ42-induced α-amylase activity was attenuated by β-adrenergic receptor antagonist propranolol. We hypothesize that astrocytes respond to fibril Aβ42 in Aβ plaques by increasing their α-amylase production to either liberate energy or regulate functions needed in reactive processes. These findings indicate α-amylase as an important actor involved in AD associated neuroinflammation.


Advancing the HIV cure agenda: the next 5 years.
To explore how ethical considerations, improved diagnostics and data from clinical trials might see the lowering of some of the barriers blocking a cure for HIV infection over the next 5 years. Despite the recent well publicized but eventually disappointing case reports, there remains only one successful HIV cure, the 'Berlin patient'. We will review the data suggesting that more potent agents might achieve in-vivo viral activation and explore the tantalizing phenomenon of 'posttreatment control' following treatment in primary HIV infection. We will also explore how new assays and novel interventions might move the field forward. There is a need for new agents that can be safely tested to impact the viral reservoir, a more meaningful understanding of how to assay patient samples, and research into mechanisms behind how the reservoir is established and impacted by therapy. With HIV+ve individuals responding so well to antiretroviral therapy, new trials must be tested hand-in-hand with guidance from patient representatives, especially with respect to determining the acceptable risk. The road to a cure is going to be difficult, but it is vital that inevitable disappointments do not detract from the final goal, which remains worth striving for.


The effect of deprivation on variations in general practitioners' referral rates: a cross sectional study of computerised data on new medical and surgical outpatient referrals in Nottinghamshire.
To determine the effect of deprivation on variations in general practitioners' referral rates using the Jarman underprivileged area (UPA(8)) score as a proxy measure. Cross sectional survey of new medical and surgical referrals from general practices to hospitals (determined from hospital activity data). All of the 183 general practices in Nottinghamshire and all of the 19 hospitals in Trent region. The relation between the referral rates per 1000 registered patients and the practice population's UPA(8) score (calculated on the basis of electoral ward), with adjustment for the number of partners, percentage of patients aged over 65 years, and fundholding status of each practice. There was a significant independent association between deprivation, as measured by the UPA(8) score, and high total referral rates and high medical referral rates (P < 0.0001). The UPA(8) score alone explained 23% of the total variation in total referral rates and 32% of the variation in medical referral rates. On multivariate analysis, where partnership size, fundholding status, and percentage of men and women aged over 65 years were included, the UPA(8) score explained 29% and 35% of the variation in total and medical referral rates respectively. Of the variables studied, the UPA(8) score was the strongest predictor of variations in referral rates. This association is most likely to be through a link with morbidity, although it could reflect differences in patients' perceptions, doctors' behaviour, or the use and provision of services.


Treatment with linezolid or vancomycin in combination with rifampin is effective in an animal model of methicillin-resistant Staphylococcus aureus foreign body osteomyelitis.
Rifampin monotherapy was compared to the combination of linezolid or vancomycin with rifampin in an experimental rat model of methicillin-resistant Staphylococcus aureus (MRSA) chronic foreign body osteomyelitis. MRSA was inoculated into the proximal tibia, and a titanium wire was implanted. Four weeks after infection, rats were treated intraperitoneally for 21 days with rifampin alone (n = 16), linezolid plus rifampin (n = 14), or vancomycin plus rifampin (n = 13). Thirteen animals received no treatment. At completion of treatment, qualitative cultures of the wire and quantitative cultures of the bone (reported as median values) were performed. Quantitative cultures from the control, rifampin monotherapy, linezolid-plus-rifampin, and vancomycin-plus-rifampin groups revealed 4.54, 0.71, 0.10, and 0.50 log₁₀ CFU/gram of bone, respectively. The bacterial load was significantly reduced in all treatment groups compared to that in the control group. Rifampin resistance was detected in isolates from 10, 2, and 1 animal in the rifampin, linezolid-plus-rifampin, and vancomycin-plus-rifampin groups, respectively. Cultures of the removed wire revealed bacterial growth in 1 and 2 animals in the rifampin and linezolid-plus-rifampin groups, respectively, with no growth in the vancomycin-plus-rifampin group and growth from all wires in the untreated group. In conclusion, we demonstrated that combination treatment with linezolid plus rifampin or vancomycin plus rifampin is effective in an animal model of MRSA foreign body osteomyelitis in the context of retention of the infected foreign body.


Experimental heatwaves negatively impact sperm quality in the zebra finch.
For sexually reproducing species, functionally competent sperm are critical to reproduction. While high atmospheric temperatures are known to influence the timing of breeding, incubation and reproductive success in birds, the effect of temperature on sperm quality remains largely unexplored. Here, we experimentally investigated the impact of ecologically relevant extreme temperatures on cloacal temperature and sperm morphology and motility in zebra finches Taeniopygia guttata We periodically sampled males exposed to 30°C or 40°C temperatures daily for 14 consecutive days. Following a 12-day (23°C) recovery period, birds were again exposed to heat, but under the alternate treatment (e.g. birds initially exposed to 40°C were exposed to 30°C). Elevated temperatures led to an increase in cloacal temperature and a reduction in the proportion of sperm with normal morphology; these effects were most notable under 40°C conditions, and were influenced by the duration of heat exposure and prior exposure to high temperature. Our findings highlight the potential role of temperature in determining male fertility in birds, and perhaps also in constraining the timing of avian breeding. Given the increased frequency of heatwaves in a warming world, our results suggest the need for further work on climatic influences on sperm quality and male fertility.


The effects of lowering LDL cholesterol with simvastatin plus ezetimibe in patients with chronic kidney disease (Study of Heart and Renal Protection): a randomised placebo-controlled trial.
Lowering LDL cholesterol with statin regimens reduces the risk of myocardial infarction, ischaemic stroke, and the need for coronary revascularisation in people without kidney disease, but its effects in people with moderate-to-severe kidney disease are uncertain. The SHARP trial aimed to assess the efficacy and safety of the combination of simvastatin plus ezetimibe in such patients. This randomised double-blind trial included 9270 patients with chronic kidney disease (3023 on dialysis and 6247 not) with no known history of myocardial infarction or coronary revascularisation. Patients were randomly assigned to simvastatin 20 mg plus ezetimibe 10 mg daily versus matching placebo. The key prespecified outcome was first major atherosclerotic event (non-fatal myocardial infarction or coronary death, non-haemorrhagic stroke, or any arterial revascularisation procedure). All analyses were by intention to treat. This trial is registered at ClinicalTrials.gov, NCT00125593, and ISRCTN54137607. 4650 patients were assigned to receive simvastatin plus ezetimibe and 4620 to placebo. Allocation to simvastatin plus ezetimibe yielded an average LDL cholesterol difference of 0·85 mmol/L (SE 0·02; with about two-thirds compliance) during a median follow-up of 4·9 years and produced a 17% proportional reduction in major atherosclerotic events (526 [11·3%] simvastatin plus ezetimibe vs 619 [13·4%] placebo; rate ratio [RR] 0·83, 95% CI 0·74-0·94; log-rank p=0·0021). Non-significantly fewer patients allocated to simvastatin plus ezetimibe had a non-fatal myocardial infarction or died from coronary heart disease (213 [4·6%] vs 230 [5·0%]; RR 0·92, 95% CI 0·76-1·11; p=0·37) and there were significant reductions in non-haemorrhagic stroke (131 [2·8%] vs 174 [3·8%]; RR 0·75, 95% CI 0·60-0·94; p=0·01) and arterial revascularisation procedures (284 [6·1%] vs 352 [7·6%]; RR 0·79, 95% CI 0·68-0·93; p=0·0036). After weighting for subgroup-specific reductions in LDL cholesterol, there was no good evidence that the proportional effects on major atherosclerotic events differed from the summary rate ratio in any subgroup examined, and, in particular, they were similar in patients on dialysis and those who were not. The excess risk of myopathy was only two per 10,000 patients per year of treatment with this combination (9 [0·2%] vs 5 [0·1%]). There was no evidence of excess risks of hepatitis (21 [0·5%] vs 18 [0·4%]), gallstones (106 [2·3%] vs 106 [2·3%]), or cancer (438 [9·4%] vs 439 [9·5%], p=0·89) and there was no significant excess of death from any non-vascular cause (668 [14·4%] vs 612 [13·2%], p=0·13). Reduction of LDL cholesterol with simvastatin 20 mg plus ezetimibe 10 mg daily safely reduced the incidence of major atherosclerotic events in a wide range of patients with advanced chronic kidney disease. Merck/Schering-Plough Pharmaceuticals; Australian National Health and Medical Research Council; British Heart Foundation; UK Medical Research Council.


Variability of coastal suprabenthic assemblages from sandy beaches of the Caribbean coast of Venezuela.
The suprabenthos or hyperbenthos is the macrofaunal assemblage of small-sized organisms that interact for some time in the benthic boundary layer. Information about the taxonomic composition and role of suprabenthic species, especially in littoral zones, is scarce and scattered. This work attempts to contribute alleviate this problem. We analyze the temporal and spatial variations of suprabenthic assemblages in the swash-zone from four beaches of the littoral coast of Venezuela. For each beach, two sites were chosen, and special attention was given to water and sediment characteristics. 12 environmental variables were measured: Dissolved oxygen, oxygen saturation percentage, pH, salinity, surface temperature, total, organic and inorganic suspended solids, total organic carbon, organic matter in sediment, grain size of sediment, and amount of dragged material of sample. All faunal samples were taken on a monthly basis during 2011; these were extracted using a manual suprabenthic sledge towed parallel to the shoreline. Samples were sorted and identified to their lowest possible taxonomic level. A total of 24 141 specimens (mean abundance: 26.16 +/- 55.35 ind./m2) belonging to 21 taxonomic groups were identified. Analysis suggests that seasonality does not explain observed changes either in fauna or environmental variables. It was found that suprabenthic assemblages, total suprabenthos density, richness and environmental variables changed in a dissimilar fashion between months and beaches. The most frequent groups were amphipods and decapods; and at the species/categories level post-larval shrimp (Penaeidae), Grapsidae crab megalopae and Arenaeus cribarius megalopae were common. Dissimilarity between months in each beach was primarily explained by the abundance of amphipods, ctenophores, decapods and mysids. For particular months and selected beaches very high abundances of ctenophores were found. This group dominated the sample even though it is not usually a representative group in suprabenthos. Samples showed low correlations between suprabenthos and environmental variables. A somewhat stronger correlation could be established between water characteristics and dragged material abundance. The studied suprabenthos assemblage was found to have high taxa richness and very dynamic behaviour at spatial and temporal scale. Further analysis suggested that there is no evident pattern of distribution and that causality can not be directly attributed to temporal variation only. Possibly there is an influence of a synergy of environmentals or biological factors, rather than a single variable. The species Americamysis bahia and Americamysis taironana are reported for the first time in Venezuela. This study represents the first ecological research of the suprabenthos in the Caribbean region.


[The influence of tobacco smoking on the lead and cadmium concentration in the urine of pregnant women and the health state of newborn].
The purpose of the study was to measure the concentration of lead and cadmium in urine of women who smoked, were exposed to environmental tobacco smoke (ETS) and were unexposed during pregnancy. The correlations between exposition to tobacco smoke and socioeconomic factors, course of pregnancy and health status of newborns were estimated. The subjects of the study were 231 pregnant women. Information about exposition to tobacco smoke, socioeconomic state, course of pregnancy and health status of newborns was collected from self-created questionnaire. The estimation of tobacco smoke exposition of the women was based on the questionnaire data and their urine cotinine concentration. The urine concentration of lead and cadmium was measured by atomic absorption spectrometry with graphite furnace. The mean urine lead concentration of women who smoked, were exposed to ETS and unexposed to tobacco smoke during pregnancy were respectively: 38.2 +/- 59.1; 36.2 +/- 50.1; 32.7 +/- 53.5 ng/ mg of creatinine, and mean cadmium concentration were respectively: 1.9 +/- 1.6; 1.1 +/- 1.2; 1.3 +/- 1.5 ng/mg of creatinine. The correlation between the urine concentration of cadmium and cotinine was statistically significant. Increased frequency of tobacco smoking was in the group of the women aged less than 25 years, that finished primary or vocational school, had monthly income less tan 500 zł/family member, and have smoking partners. In the group of women who smoked during pregnancy the childbirth was statistically more often by caesarian and supported delivery. The newborns of mothers who smoked and were exposed to ETS had birth weight respectively 348.5 g and 281.1 g smaller than newborns of unexposed mothers. The length of newborns of smoking and ETS exposed mothers were respectively 2.8 cm and 0.7 cm shorter then newborns of unexposed mothers.


Imaging and Impact of Myocardial Fibrosis in Aortic Stenosis.
Aortic stenosis is characterized both by progressive valve narrowing and the left ventricular remodeling response that ensues. The only effective treatment is aortic valve replacement, which is usually recommended in patients with severe stenosis and evidence of left ventricular decompensation. At present, left ventricular decompensation is most frequently identified by the development of typical symptoms or a marked reduction in left ventricular ejection fraction <50%. However, there is growing interest in using the assessment of myocardial fibrosis as an earlier and more objective marker of left ventricular decompensation, particularly in asymptomatic patients, where guidelines currently rely on nonrandomized data and expert consensus. Myocardial fibrosis has major functional consequences, is the key pathological process driving left ventricular decompensation, and can be divided into 2 categories. Replacement fibrosis is irreversible and identified using late gadolinium enhancement on cardiac magnetic resonance, while diffuse fibrosis occurs earlier, is potentially reversible, and can be quantified with cardiac magnetic resonance T1 mapping techniques. There is a substantial body of observational data in this field, but there is now a need for randomized clinical trials of myocardial imaging in aortic stenosis to optimize patient management. This review will discuss the role that myocardial fibrosis plays in aortic stenosis, how it can be imaged, and how these approaches might be used to track myocardial health and improve the timing of aortic valve replacement.


Utilizing High-Density Electroencephalography and Motion Capture Technology to Characterize Sensorimotor Integration While Performing Complex Actions.
Studies of sensorimotor integration often use sensory stimuli that require a simple motor response, such as a reach or a grasp. Recent advances in neural recording techniques, motion capture technologies, and time-synchronization methods enable studying sensorimotor integration using more complex sensory stimuli and performed actions. Here, we demonstrate that prehensile actions that require using complex sensory instructions for manipulating different objects can be characterized using high-density electroencephalography and motion capture systems. In 20 participants, we presented stimuli in different sensory modalities (visual, auditory) containing different contextual information about the object with which to interact. Neural signals recorded near motor cortex and posterior parietal cortex discharged based on both the instruction delivered and object manipulated. Additionally, kinematics of the wrist movements could be discriminated between participants. These findings demonstrate a proof-of-concept behavioral paradigm for studying sensorimotor integration of multidimensional sensory stimuli to perform complex movements. The designed framework will prove vital for studying neural control of movements in clinical populations in which sensorimotor integration is impaired due to information no longer being communicated correctly between brain regions (e.g. stroke). Such a framework is the first step towards developing a neural rehabilitative system for restoring function more effectively.


Engineering strategy to improve peptide analogs: from structure-based computational design to tumor homing.
We present a chemical strategy to engineer analogs of the tumor-homing peptide CREKA (Cys-Arg-Glu-Lys-Ala), which binds to fibrin and fibrin-associated clotted plasma proteins in tumor vessels (Simberg et al. in Proc Natl Acad Sci USA 104:932-936, 2007) with improved ability to inhibit tumor growth. Computer modeling using a combination of simulated annealing and molecular dynamics were carried out to design targeted replacements aimed at enhancing the stability of the bioactive conformation of CREKA. Because this conformation presents a pocket-like shape with the charged groups of Arg, Glu and Lys pointing outward, non-proteinogenic amino acids α-methyl and N-methyl derivatives of Arg, Glu and Lys were selected, rationally designed and incorporated into CREKA analogs. The stabilization of the bioactive conformation predicted by the modeling for the different CREKA analogs matched the tumor fluorescence results, with tumor accumulation increasing with stabilization. Here we report the modeling, synthetic procedures, and new biological assays used to test the efficacy and utility of the analogs. Combined, our results show how studies based on multi-disciplinary collaboration can converge and lead to useful biomedical advances.


Isolation and identification of bacteria from activated sludge purifying petroleum wastewaters.
Several strains growing well in minimal media with 500 and 1000 mg/l of oil or phenol as a sole carbon source were isolated from activated sludge purifying petroleum waste waters and identified. Five of the best growing strains classified as Arthrobacter, Pseudomonas, Xanthomonas and Enterobacter were selected and their capacity to remove petroleum components and phenol (in the oil fraction of petrochemical waste waters) was studied. The enzymatic activity of the strains, including respiration intensity and dehydrogenase activity was also determined. All the examined strains were found to use oils as the sole source of carbon (percent age of the oils reduction during cultivation of the individual strains ranged from 58 to 78). Phenol was completely reduced by only one strain. The rest of the strain removed only from 7 to 24% of this compound. The activity of dehydrogenases and the respiration intensity in the presence of the studied substrates -- oil and phenol was low for all the examined strains.


New insight into the structural evolution of PbTiO3: an unbiased structure search.
Understanding the structural evolution of materials is a challenging problem of condensed matter physics. Solving this problem would open new ways for understanding the behaviors of materials. In this context, we here report unbiased structure searches for a prototypical perovskite oxide, PbTiO3, based on the CALYPSO (Crystal structure AnaLYsis by Particle Swarm Optimization) method in conjunction with first-principles calculations. For the first time, we decipher the structure evolution of PbTiO3 from a zero dimensional (0D) cluster to a two dimensional (2D) layered structure and in the end to a three dimensional (3D) bulk solid. Our unbiased structure search is successful in reproducing the cubic Pm3[combining macron]m and tetragonal P4mm phases of PbTiO3 at ambient pressure. We also predict a new quasi-planar kite shape structure of the PbTiO3 cluster, with Cs symmetry and a surprisingly large HOMO-LUMO gap. Furthermore, by using this method, we predict that the 2D planar PbTiO3 monolayer is unstable in the perpendicular direction and the 2D PbTiO3 double layer is dynamically stable, with a hope that it can provide guidance to future synthesis of low dimensional perovskite oxides.


Measles virus selectively blind to signaling lymphocytic activation molecule (SLAM; CD150) is attenuated and induces strong adaptive immune responses in rhesus monkeys.
The signaling lymphocytic activation molecule (SLAM; CD150) is the immune cell receptor for measles virus (MV). To assess the importance of the SLAM-MV interactions for virus spread and pathogenesis, we generated a wild-type IC-B MV selectively unable to recognize human SLAM (SLAM-blind). This virus differs from the fully virulent wild-type IC-B strain by a single arginine-to-alanine substitution at amino acid 533 of the attachment protein hemagglutinin and infects cells through SLAM about 40 times less efficiently than the isogenic wild-type strain. Ex vivo, this virus infects primary lymphocytes at low levels regardless of SLAM expression. When a group of six rhesus monkeys (Macaca mulatta) was inoculated intranasally with the SLAM-blind virus, no clinical symptoms were documented. Only one monkey had low-level viremia early after infection, whereas all the hosts in the control group had high viremia levels. Despite minimal, if any, viremia, all six hosts generated neutralizing antibody titers close to those of the control monkeys while MV-directed cellular immunity reached levels at least as high as in wild-type-infected monkeys. These findings prove formally that efficient SLAM recognition is necessary for MV virulence and pathogenesis. They also suggest that the selectively SLAM-blind wild-type MV can be developed into a vaccine vector.


Nursing home 5-star rating system exacerbates disparities in quality, by payer source.
Market-based reforms in health care, such as public reporting of quality, may inadvertently exacerbate disparities. We examined how the Centers for Medicare and Medicare Services' five-star rating system for nursing homes has affected residents who are dually enrolled in Medicare and Medicaid ("dual eligibles"), a particularly vulnerable and disadvantaged population. Specifically, we assessed the extent to which dual eligibles and non-dual eligibles avoided the lowest-rated nursing homes and chose the highest-rated homes once the five-star rating system began, in late 2008. We found that both populations resided in better-quality homes over time but that by 2010 the increased likelihood of choosing the highest-rated homes was substantially smaller for dual eligibles than for non-dual eligibles. Thus, the gap in quality, as measured by a nursing home's star rating, grew over time. Furthermore, we found that the benefit of the five-star system to dual eligibles was largely due to providers' improving their ratings, not to consumers' choosing different providers. We present evidence suggesting that supply constraints play a role in limiting dual eligibles' responses to quality ratings, since high-quality providers tend to be located close to relatively affluent areas. Increases in Medicaid payment rates for nursing home services may be the only long-term solution.


The impact of alcohol on HIV prevention and treatment for South Africans in primary healthcare.
Antiretroviral treatment (ART) has substantially reduced morbidity and mortality for HIV patients. In South Africa, with the largest ART programme globally, attention is needed not only on the further expansion of ART coverage, but also on factors which undermine its effectiveness, such as alcohol use. Given the decentralised approach of nurse-initiated and -sustained ART in the South African primary health sector, it is important to document key aspects of alcohol use to be conveyed to HIV-positive individuals and those at risk for HIV. This study comprised a narrative review of relevant literature. Alcohol acts through both behavioural and physiological pathways to impact on the acquisition, further transmission and then progression of HIV disease. Besides links to risky sex, alcohol undermines the immune system, raising susceptibility to contracting and then countering HIV and other infections. There are important drug interactions between alcohol and ART, or therapies for opportunistic infections and other co-morbidities. Moreover, alcohol undermines adherence to the medication which is essential for effective ART. Primary healthcare clinic attendees need evidence-based information on the detrimental effects of alcohol consumption on HIV infection, which ensue throughout the clinical course of HIV. This spans the role of alcohol consumption as a risk factor for HIV infection, HIV replication in infected individuals, a person's response to HIV infection and HIV treatment. Primary healthcare workers, especially nurses and HIV counsellors, require training in order to screen for and provide appropriate interventions for HIV-positive patients, those on treatment and treatment-naïve patients, who will benefit from reduced alcohol consumption or the cessation thereof.


Daily interpersonal events in pain patients: applying action theory to chronic illness.
Action theory proposes that individuals actively shape and then respond to their environments, highlighting the role of stable person characteristics in the development and maintenance of life's interpersonal difficulties. In this study, the authors adopt the action perspective in their examination of predictors of daily interpersonal events among chronic pain patients with rheumatoid arthritis. They probe the extent to which stable symptoms of illness explained between-person variation, and fluctuating symptoms explain day-to-day variation in both positive and negative events. Their evaluation of patients' daily diary reports indicate that between-person differences accounted for more variance in the occurrence of positive events relative to negative events (48% vs. 31%, respectively). Likewise, between-person factors accounted for more variance in appraisals of positive compared to negative events across relationship domains. Both intractable illness symptoms and disability, and daily fluctuations in pain and fatigue, were only weakly related to patients' reports of their interpersonal experiences. Consistent with action theory, these results suggest that stable person characteristics are strongly related to daily stressors and particularly daily positive events in pain patients, but still account for less than 50% of the variance in events and their appraisals. In contrast, elevations in illness-related features, both between individuals and within individuals from day-to-day, are not robust predictors of positive or negative social exchanges. These findings point to the value of capturing the experiences of individuals intensively over time, an approach that can help to elaborate the contributions of both stable factors and circumstance in shaping social contexts in chronic illness.


The skeleton of postmetamorphic echinoderms in a changing world.
Available evidence on the impact of acidification and its interaction with warming on the skeleton of postmetamorphic (juvenile and adult) echinoderms is reviewed. Data are available on sea urchins, starfish, and brittle stars in 33 studies. Skeleton growth of juveniles of all sea urchin species studied so far is affected from pH 7.8 to 7.6 in seawater, values that are expected to be reached during the 21st century. Growth in adult sea urchins (six species studied) is apparently only marginally affected at seawater pH relevant to this century. The interacting effect of temperature differed according to studies. Juvenile starfish as well as adults seem to be either not impacted or even boosted by acidification. Brittle stars show moderate effects at pH below or equal to 7.4. Dissolution of the body wall skeleton is unlikely to be a major threat to sea urchins. Spines, however, due to their exposed position, are more prone to this threat, but their regeneration abilities can probably ensure their maintenance, although this could have an energetic cost and induce changes in resource allocation. No information is available on skeleton dissolution in starfish, and the situation in brittle stars needs further assessment. Very preliminary evidence indicates that mechanical properties in sea urchins could be affected. So, although the impact of ocean acidification on the skeleton of echinoderms has been considered as a major threat from the first studies, we need a better understanding of the induced changes, in particular the functional consequences of growth modifications and dissolution related to mechanical properties. It is suggested to focus studies on these aspects.


Evaluation of microsatellite instability and immunohistochemistry for the prediction of germ-line MSH2 and MLH1 mutations in hereditary nonpolyposis colon cancer families.
Forty-eight hereditary nonpolyposis colorectal carcinoma (HNPCC) families for which a tumor sample was available were evaluated for the presence of germ-line mutations in MSH2 and MLH1, tumor microsatellite instability (MSI), and where possible, expression of MSH2 and MLH1 in tumors by immunohistochemistry. Fourteen of 48 of the families had a germ-line mutation in either MSH2 or MLH1 that could be detected by genomic DNA sequencing, and 28 of 48 of the families had MSI-H tumors. Four additional families showed loss of expression of MSH2, and one additional family showed loss of expression of MLH1 but did not have germ-line mutations in MSH2 or MLH1 that could be detected by DNA sequencing. MSI-H, as defined using the National Cancer Institute recommended five-microsatellite panel, had a 100% sensitivity for identifying samples having MSH2 or MLH1 mutations or loss of expression. In contrast, loss of MSH2 and MLH1 expression did not identify all samples having germ-line mutations in MSH2 or MLH1, because in five cases, a mutant protein product was expressed that could be detected by IHC. A combination of the Bethesda criteria for HNPCC and an MSI-H phenotype defined the smallest number of cases having all of the germ-line MSH2 and MLH1 mutations that could be detected by DNA sequencing.


Robust laser speckle recognition system for authenticity identification.
This paper proposes a laser speckle recognition system for authenticity verification. Because of the unique imperfection surfaces of objects, laser speckle provides identifiable features for authentication. A Gabor filter, SIFT (Scale-Invariant Feature Transform), and projection were used to extract the features of laser speckle images. To accelerate the matching process, the extracted Gabor features were organized into an indexing structure using the K-means algorithm. Plastic cards were used as the target objects in the proposed system and the hardware of the speckle capturing system was built. The experimental results showed that the retrieval performance of the proposed method is accurate when the database contains 516 laser speckle images. The proposed system is robust and feasible for authenticity verification.


Thymoquinone: an emerging natural drug with a wide range of medical applications.
Nigella sativa has attracted healers in ancient civilizations and researchers in recent times. Traditionally, it has been used in different forms to treat many diseases including asthma, hypertension, diabetes, inflammation, cough, bronchitis, headache, eczema, fever, dizziness and influenza. Experimentally, it has been demonstrated that N. sativa extracts and the main constituent of their volatile oil, thymoquinone, possess antioxidant, anti-inflammatory and hepatoprotective properties. In this review we aimed at summarizing the most recent investigations related to a few and most important effects of thymoquinone. It is concluded that thymoquinone has evidently proved its activity as hepatoprotective, anti-inflammatory, antioxidant, cytotoxic and anti-cancer chemical, with specific mechanisms of action, which provide support to consider this compound as an emerging drug. Further research is required to make thymoquinone a pharmaceutical preparation ready for clinical trials.


Breastfeeding peer supporters and a community support group: evaluating their effectiveness.
Peer support for breastfeeding mothers has often been promoted as a way of increasing rates in communities with low breastfeeding prevalence, where there are few breastfeeding role models and a bottle-feeding culture predominates. This study evaluated the effectiveness of a peer support initiative, which trained peer supporters who then set up a support group, in an area of social and economic deprivation in South Bristol, UK. The effects of training on 6 local women were explored; the combined effect of peer support and the group on breastfeeding prevalence assessed; and mothers' views on what they had gained from the breastfeeding support group reported. Focus groups were held with the peer supporters, who also completed questionnaires before and after their training. Questionnaires were sent to all mothers (35) who attended the support group over the first 5 months to ascertain their views. Breastfeeding rates were extracted from routinely collected sources for both the local area and the wider Bristol population (excluding the peer support area). The initial training increased the peer supporters' knowledge about breastfeeding and their confidence in talking to and supporting mothers. The number of mothers attending the breastfeeding support group increased from 3 per week in May to 10 per week by September 2001. Mothers who attended the group particularly appreciated being able to talk about breastfeeding and getting consistent breastfeeding advice. Breastfeeding rates in the area for 12-months before and after the start of the peer support initiative showed a significant increase at 8 weeks (7%), which was higher than the overall increase in the wider Bristol area (3%). This evaluation suggests that peer supporters combined with a breastfeeding support group are an effective way of increasing breastfeeding prevalence in areas of low continuation.


Traditional agricultural practices and the sex ratio today.
We study the historical origins of cross-country differences in the male-to-female sex ratio. Our analysis focuses on the use of the plough in traditional agriculture. In societies that did not use the plough, women tended to participate in agriculture as actively as men. By contrast, in societies that used the plough, men specialized in agricultural work, due to the physical strength needed to pull the plough or control the animal that pulls it. We hypothesize that this difference caused plough-using societies to value boys more than girls. Today, this belief is reflected in male-biased sex ratios, which arise due to sex-selective abortion or infanticide, or gender-differences in access to family resources, which results in higher mortality rates for girls. Testing this hypothesis, we show that descendants of societies that traditionally practiced plough agriculture today have higher average male-to-female sex ratios. We find that this effect systematically increases in magnitude and statistical significance as one looks at older cohorts. Estimates using instrumental variables confirm our findings from multivariate OLS analysis.


The deletion of several amino acid stretches of Escherichia coli alpha-hemolysin (HlyA) suggests that the channel-forming domain contains beta-strands.
Escherichia coli α-hemolysin (HlyA) is a pore-forming protein of 110 kDa belonging to the family of RTX toxins. A hydrophobic region between the amino acid residues 238 and 410 in the N-terminal half of HlyA has previously been suggested to form hydrophobic and/or amphipathic α-helices and has been shown to be important for hemolytic activity and pore formation in biological and artificial membranes. The structure of the HlyA transmembrane channel is, however, largely unknown. For further investigation of the channel structure, we deleted in HlyA different stretches of amino acids that could form amphipathic β-strands according to secondary structure predictions (residues 71-110, 158-167, 180-203, and 264-286). These deletions resulted in HlyA mutants with strongly reduced hemolytic activity. Lipid bilayer measurements demonstrated that HlyAΔ71-110 and HlyAΔ264-286 formed channels with much smaller single-channel conductance than wildtype HlyA, whereas their channel-forming activity was virtually as high as that of the wildtype toxin. HlyAΔ158-167 and HlyAΔ180-203 were unable to form defined channels in lipid bilayers. Calculations based on the single-channel data indicated that the channels generated by HlyAΔ71-110 and HlyAΔ264-286 had a smaller size (diameter about 1.4 to 1.8 nm) than wildtype HlyA channels (diameter about 2.0 to 2.6 nm), suggesting that in these mutants part of the channel-forming domain was removed. Osmotic protection experiments with erythrocytes confirmed that HlyA, HlyAΔ71-110, and HlyAΔ264-286 form defined transmembrane pores and suggested channel diameters that largely agreed with those estimated from the single-channel data. Taken together, these results suggest that the channel-forming domain of HlyA might contain β-strands, possibly in addition to α-helical structures.


Psychological support for patients with cancer: evidence review and suggestions for future directions.
Psychological distress and mental health comorbidity are common in cancer. Various therapeutic frameworks have been used for interventions to improve psychological wellbeing and quality of life in cancer patients with mixed results. This article reviews contributions to that literature published since January 2017. The majority of new psychological intervention research in cancer has used cognitive behavioural therapy (CBT) or mindfulness-based interventions. CBT has been considered a gold-standard intervention and recent evidence justifies continuation of this. Recent reviews call into question the validity of evidence for mindfulness-based interventions. A smaller number of trials using acceptance and commitment therapy, meta-cognitive therapy, dignity therapy and coaching have emerged, and whereas findings are promising, additional fully powered trials are required. Weaker evidence exists for counselling, support-based and narrative therapy interventions. Efficacious, timely and acceptable psychological interventions are a necessary component of comprehensive cancer care. There is some way to go before the evidence conclusively points towards which interventions work for which cancer groups and for which specific outcomes. Methodological limitations must be addressed in future trials; at the forefront remains the need for fully powered, head-to-head comparison trials.


Phishing simulation exercise in a large hospital: A case study.
Phishing is a major threat to the data and infrastructure of healthcare organizations and many cyberattacks utilize this socially engineered pathway. Phishing simulation is used to identify weaknesses and risks in the human defences of organizations. There are many factors influencing the difficulty of detecting a phishing email including fatigue and the nature of the deceptive message. A major Italian Hospital with over 6000 healthcare staff performed a phishing simulation as part of its annual training and risk assessment. Three campaigns were launched at approx. 4-month intervals, to compare staff reaction to a general phishing email and a customized one. The results show that customization of phishing emails makes them much more likely to be acted on. In the first campaign, 64% of staff did not open the general phish, significantly more than the 38% that did not open the custom phish. A significant difference was also found for the click rate, with significantly more staff clicking on the custom phish. However, the campaigns could not be run as intended, due to issues raised within the organization. Phishing simulation is useful but not without its limitations. It requires contextual knowledge, skill and experience to ensure that it is effective. The exercise raised many issues within the Hospital. Successful, ethical phishing simulations require coordination across the organization, precise timing and lack of staff awareness. This can be complex to coordinate. Misleading messages containing false threats or promises can cause a backlash from staff and unions. The effectiveness of the message is dependent on the personalization of the message to current, local events. The lessons learned can be useful for other hospitals.


Defending plasma T3 is a biological priority.
Triiodothyronine (T3), the active form of thyroid hormone is produced predominantly outside the thyroid parenchyma secondary to peripheral tissue deiodination of thyroxine (T4), with <20% being secreted directly from the thyroid. In healthy individuals, plasma T3 is regulated by the negative feedback loop of the hypothalamus-pituitary-thyroid axis and by homoeostatic changes in deiodinase expression. Therefore, with the exception of a minimal circadian rhythmicity, serum T3 levels are stable over long periods of time. Studies in rodents indicate that different levels of genetic disruption of the feedback mechanism and deiodinase system are met with increase in serum T4 and thyroid-stimulating hormone (TSH) levels, while serum T3 levels remain stable. These findings have focused attention on serum T3 levels in patients with thyroid disease, with important clinical implications affecting therapeutic goals and choice of therapy for patients with hypothyroidism. Although monotherapy with levothyroxine is the standard of care for hypothyroidism, not all patients normalize serum T3 levels with many advocating for combination therapy with levothyroxine and liothyronine. The latter could be relevant for a significant number of patients that remain symptomatic on monotherapy with levothyroxine, despite normalization of serum TSH levels.


Pericapillary basement membrane thickening in human skeletal muscles.
The basement membrane (BM) surrounding capillaries in skeletal muscles varies physiologically in thickness according to age, physical fitness, and anatomical site in humans. Furthermore, the pericapillary BM thickness (CBMT) increases pathophysiologically during several common disease states, including peripheral arterial disease and diabetes mellitus. This review on CBM thickening in human skeletal muscles is two pronged. First, it addresses the advantages/disadvantages of grid- and tablet-based measuring and morphometric techniques that are implemented to assess the CBMT on transmission electron micrographs. Second, it deals with the biology of CBM thickening in skeletal muscles, particularly its possible causes, molecular mechanisms, and functional impact. CBM thickening is triggered by several physical factors, including diabetes-associated glycation, hydrostatic pressure, and inflammation. Increased biosynthesis of type IV collagen expression or repetitive cycles in pericyte or endothelial cell degeneration/proliferation appear to be most critical for CBM accumulation. A thickened CBM obviously poses a greater barrier for diffusion, lowers the microvascular elasticity, and impedes transcytosis of inflammatory cells. Our own morphometric data reveal the CBM enlargement to be not accompanied by the pericyte coverage. Owing to an overlap or redundancy in the capillary supply, CBM thickening in skeletal muscles might not be such a devastating occurrence as in organs with endarterial circulation (e.g., kidney and retina). CBM growth in skeletal muscles can be reversed by training or administration of antidiabetic drugs. In conclusion, CBM thickening in skeletal muscles is a microvascular remodeling process by which metabolic, hemodynamic, and inflammatory forces are integrated together and which could play a hitherto underestimated role in etiology/progression of human diseases.


Investigating Alfvénic wave propagation in coronal open-field regions.
The physical mechanisms behind accelerating solar and stellar winds are a long-standing astrophysical mystery, although recent breakthroughs have come from models invoking the turbulent dissipation of Alfvén waves. The existence of Alfvén waves far from the Sun has been known since the 1970s, and recently the presence of ubiquitous Alfvénic waves throughout the solar atmosphere has been confirmed. However, the presence of atmospheric Alfvénic waves does not, alone, provide sufficient support for wave-based models; the existence of counter-propagating Alfvénic waves is crucial for the development of turbulence. Here, we demonstrate that counter-propagating Alfvénic waves exist in open coronal magnetic fields and reveal key observational insights into the details of their generation, reflection in the upper atmosphere and outward propagation into the solar wind. The results enhance our knowledge of Alfvénic wave propagation in the solar atmosphere, providing support and constraints for some of the recent Alfvén wave turbulence models.


Oleuropein, the Main Polyphenol of Olea europaea Leaf Extract, Has an Anti-Cancer Effect on Human BRAF Melanoma Cells and Potentiates the Cytotoxicity of Current Chemotherapies.
Oleuropein (Ole), a secoiridoid glucoside present in Olea europaea leaves, gained scientific interest thanks to its several biological properties, including the anticancer one. We verified whether Ole might potentiate the cytotoxicity of conventional drugs used to treat melanoma, disclosing a potentially new therapeutic strategy. We tested the cytotoxic action of Ole alone or in combination with chemotherapeutics on A375 human melanoma cells. We found that Ole was able, at a dose of 500 µM, to stimulate apoptosis, while at a non-toxic dose of 250 µM, it affected cell proliferation and induced the downregulation of the pAKT/pS6 pathway. A dose of 250 µM Ole did not potentiate the effect of Vemurafenib (PLX4032), but it succeeded in increasing the cytotoxic effect of Dacarbazine (DTIC). The major effect was found in the association between Ole and Everolimus (RAD001), also on PLX4032-resistant BRAF melanoma cells, which possibly cooperate in the inhibition of the pAKT/pS6 pathway. Of interest, an olive leaf extract enriched in equimolar Ole was more effective and able to further improve DTIC and RAD001 efficacy on BRAF melanoma cells with respect to Ole alone. Therefore, Ole represents a natural product able to potentiate a wide array of chemotherapeutics against BRAF melanoma cells affecting the pAKT/pS6 pathway.


A new mixture model approach to analyzing allelic-loss data using Bayes factors.
Allelic-loss studies record data on the loss of genetic material in tumor tissue relative to normal tissue at various loci along the genome. As the deletion of a tumor suppressor gene can lead to tumor development, one objective of these studies is to determine which, if any, chromosome arms harbor tumor suppressor genes. We propose a large class of mixture models for describing the data, and we suggest using Bayes factors to select a reasonable model from the class in order to classify the chromosome arms. Bayes factors are especially useful in the case of testing that the number of components in a mixture model is n0 versus n1. In these cases, frequentist test statistics based on the likelihood ratio statistic have unknown distributions and are therefore not applicable. Our simulation study shows that Bayes factors favor the right model most of the time when tumor suppressor genes are present. When no tumor suppressor genes are present and background allelic-loss varies, the Bayes factors are often inconclusive, although this results in a markedly reduced false-positive rate compared to that of standard frequentist approaches. Application of our methods to three data sets of esophageal adenocarcinomas yields interesting differences from those results previously published. Our results indicate that Bayes factors are useful for analyzing allelic-loss data.


Anger Expression Types and Interpersonal Problems in Nurses.
The purpose of this study was to investigate the anger expression types in nurses and to analyze the differences between the anger expression types and interpersonal problems. The data were collected from 149 nurses working in general hospitals with 300 beds or more in Seoul or Gyeonggi province, Korea. For anger expression type, the anger expression scale from the Korean State-Trait Anger Expression Inventory was used. For interpersonal problems, the short form of the Korean Inventory of Interpersonal Problems Circumplex Scales was used. Data were analyzed using descriptive statistics, cluster analysis, multivariate analysis of variance, and Duncan's multiple comparisons test. Three anger expression types in nurses were found: low-anger expression, anger-in, and anger-in/control type. From the results of multivariate analysis of variance, there were significant differences between anger expression types and interpersonal problems (Wilks lambda F = 3.52, p < .001). Additionally, anger-in/control type was found to have the most difficulty with interpersonal problems by Duncan's post hoc test (p < .050). Based on this research, the development of an anger expression intervention program for nurses is recommended to establish the means of expressing the suppressed emotions, which would help the nurses experience less interpersonal problems.


Mapping Research Domain Criteria using a transdiagnostic mini-RDoC assessment in mental disorders: a confirmatory factor analysis.
This study aimed to build on the relationship of well-established self-report and behavioral assessments to the latent constructs positive (PVS) and negative valence systems (NVS), cognitive systems (CS), and social processes (SP) of the Research Domain Criteria (RDoC) framework in a large transnosological population which cuts across DSM/ICD-10 disorder criteria categories. One thousand four hundred and thirty one participants (42.1% suffering from anxiety/fear-related, 18.2% from depressive, 7.9% from schizophrenia spectrum, 7.5% from bipolar, 3.4% from autism spectrum, 2.2% from other disorders, 18.4% healthy controls, and 0.2% with no diagnosis specified) recruited in studies within the German research network for mental disorders for the Phenotypic, Diagnostic and Clinical Domain Assessment Network Germany (PD-CAN) were examined with a Mini-RDoC-Assessment including behavioral and self-report measures. The respective data was analyzed with confirmatory factor analysis (CFA) to delineate the underlying latent RDoC-structure. A revised four-factor model reflecting the core domains positive and negative valence systems as well as cognitive systems and social processes showed a good fit across this sample and showed significantly better fit compared to a one factor solution. The connections between the domains PVS, NVS and SP could be substantiated, indicating a universal latent structure spanning across known nosological entities. This study is the first to give an impression on the latent structure and intercorrelations between four core Research Domain Criteria in a transnosological sample. We emphasize the possibility of using already existing and well validated self-report and behavioral measurements to capture aspects of the latent structure informed by the RDoC matrix.


The psychoactive aminoalkylbenzofuran derivatives, 5-APB and 6-APB, mimic the effects of 3,4-methylenedioxyamphetamine (MDA) on monoamine transmission in male rats.
The nonmedical use of new psychoactive substances (NPS) is a worldwide public health concern. The so-called "benzofury" compounds, 5-(2-aminopropyl)benzofuran (5-APB) and 6-(2-aminopropyl)benzofuran (6-APB), are NPS with stimulant-like properties in human users. These substances are known to interact with monoamine transporters and 5-HT receptors in transfected cells, but less is known about their effects in animal models. Here, we used in vitro monoamine transporter assays in rat brain synaptosomes to characterize the effects of 5-APB and 6-APB, together with their N-methyl derivatives 5-MAPB and 6-MAPB, in comparison with 3,4-methylenedioxyamphetamine (MDA) and 3,4-methylenedioxymethamphetamine (MDMA). In vivo neurochemical and behavioral effects of 5-APB (0.3 and 1.0 mg/kg, i.v.) and 6-APB (0.3 and 1.0 mg/kg, i.v.) were assessed in comparison with MDA (1.0 and 3.0 mg/kg, i.v.) using microdialysis sampling in the nucleus accumbens of conscious male rats. All four benzofuran derivatives were substrate-type releasers at dopamine transporters (DAT), norepinephrine transporters (NET), and serotonin transporters (SERT) with nanomolar potencies, similar to the profile of effects produced by MDA and MDMA. However, the benzofurans were at least threefold more potent than MDA and MDMA at evoking transporter-mediated release. Like MDA, both benzofurans induced dose-related elevations in extracellular dopamine and serotonin in the brain, but benzofurans were more potent than MDA. The benzofuran derivatives also induced profound behavioral activation characterized by forward locomotion which lasted for at least 2 h post-injection. Overall, benzofurans are more potent than MDA in vitro and in vivo, producing sustained stimulant-like effects in rats. These data suggest that benzofuran-type compounds may have abuse liability and could pose risks for adverse effects, especially if used in conjunction with abused drugs or medications which enhance monoamine transmission in the brain.


Variability Among Forecast Models for the Apple Sooty Blotch/Flyspeck Disease Complex.
Several disease forecast models have been developed to guide treatment of the sooty blotch and flyspeck (SBFS) disease complex of apple. Generally, these empirical models are based on the accumulation of hours of leaf wetness (leaf wetness duration [LWD]) from a biofix at or near the phenological growth stage petal fall, when apple flower petals senesce and drop. The models recommend timing of the initial fungicide application targeting SBFS. However, there are significant differences among SBFS forecast models in terms of biofix and the length of LWD thresholds. A comparison of models using a single input data set generated recommendations for the first SBFS fungicide application that differed by up to 5 weeks. In an attempt to improve consistency among models, potential sources for differences were examined. Leaf wetness (LW) is a particularly variable parameter among models, depending on whether on-site or remote weather data were used, the types of sensors and their placement for on-site monitors, and the models used to estimate LW remotely. When SBFS models are applied in the field, recommended treatment thresholds do not always match the method of data acquisition, leading to potential failures. Horticultural factors, such as tree size, canopy density, and cultivar, and orchard site factors such as the distance to potential inoculum sources can impact risk of SBFS and should also be considered in forecast models. The number of fungal species identified as contributors to the SBFS disease complex has expanded tremendously in recent years. A lack of understanding of key epidemiological factors for different fungi in the complex, and which fungi represent the most challenging management problems, are obvious issues in the development of improved SBFS models. If SBFS forecast models are to be adopted, researchers will need to address these issues.


simuPOP: a forward-time population genetics simulation environment.
simuPOP is a forward-time population genetics simulation environment. The core of simuPOP is a scripting language (Python) that provides a large number of objects and functions to manipulate populations, and a mechanism to evolve populations forward in time. Using this R/Splus-like environment, users can create, manipulate and evolve populations interactively, or write a script and run it as a batch file. Owing to its flexible and extensible design, simuPOP can simulate large and complex evolutionary processes with ease. At a more user-friendly level, simuPOP provides an increasing number of built-in scripts that perform simulations ranging from implementation of basic population genetics models to generating datasets under complex evolutionary scenarios. simuPOP is freely available at http://simupop.sourceforge.net, distributed under GPL license.


Outcomes of phacoemulsification in patients with chronic ocular graft-versus-host disease.
The purpose of this study was to evaluate the outcomes of phacoemulsification in patients with ocular graft-versus-host disease (GVHD). The occurrence of cataracts, cataract surgery, and its outcomes were analyzed in the medical records of 229 patients (458 eyes) with ocular GVHD. Outcome measures included pre- and postoperative corrected distance visual acuity (CDVA) and the rate of postoperative complications. Of the 458 eyes evaluated, 58 were pseudophakic; from the 400 phakic eyes, 238 (59 %) presented with cataracts and 62 (26 %) underwent cataract surgery. Analysis of postoperative complications and visual outcomes at 1 month was performed in 51 eyes in which detailed surgical and immediate postoperative records were available. Preoperatively, the mean CDVA was 0.67 ± 0.57 LogMAR (Snellen 20/93), improving postoperatively to 0.17 ± 0.18 (Snellen 20/29) at 1 month (P < 0.0001), and to 0.13 ± 0.14 (Snellen 20/26) by the final follow-up visit (P < 0.0001). Postoperative complications included corneal epithelial defects (8 %), filamentary keratitis (6 %), worsening of corneal epitheliopathy (16 %), posterior capsular opacification (18 %), and cystoid macular edema (4 %). A corrected distance visual acuity of 20/30 or better was achieved in 87 % of the eyes; suboptimal CDVA improvement was attributable to severe ocular surface disease, pre-existing advanced glaucoma, and prior macular surgery. Phacoemulsification in patients with chronic ocular GVHD is a safe and efficacious procedure resulting in significant visual improvement. Overall, postoperative adverse events responded well to timely management.


Rad6-dependent ubiquitination of histone H2B in yeast.
Although ubiquitinated histones are present in substantial levels in vertebrate cells, the roles they play in specific biological processes and the cellular factors that regulate this modification are not well characterized. Ubiquitinated H2B (uH2B) has been identified in the yeast Saccharomyces cerevisiae, and mutation of the conserved ubiquitination site is shown to confer defects in mitotic cell growth and meiosis. uH2B was not detected in rad6 mutants, which are defective for the ubiquitin-conjugating enzyme Ubc2, thus identifying Rad6 as the major cellular activity that ubiquitinates H2B in yeast.


Low-Income Children With Chronic Conditions Face Increased Costs If Shifted From CHIP To Marketplace Plans.
More than eight million children risk having their health insurance coverage disrupted if federal funding for the Children's Health Insurance Program (CHIP) is not extended beyond 2017. In this study we explored two current policy alternatives: extending federal funding for CHIP or enrolling children in the existing health insurance Marketplace plans. We simulated annual out-of-pocket expenses using detailed health plan data from CHIP and federally facilitated Marketplace plans for a nationally representative cohort of children with chronic conditions, conducting comparisons at four different percentage categories of the federal poverty level. If CHIP funding is not renewed and children with chronic conditions shift to coverage under Marketplace plans, their families face increased annual out-of-pocket expenses ranging from $233 at the lowest income levels to $2,472 at the highest income level of 251-400 percent of poverty. Families with children who have epilepsy, diabetes, or mood disorders may face the highest costs. Cost sharing for prescription drugs (25 percent) and hospitalizations (23 percent) account for much of the difference. Absent enhancements to Marketplace cost-sharing protections, and given recent efforts to repeal the Affordable Care Act, renewing funding for CHIP will provide the greatest financial protections to families of income-eligible children with chronic conditions.


Genetic association between alleles of pancreatic phospholipase A2 gene and bipolar affective disorder.
Chromosome 12q is a region of interest for the genetics of bipolar affective disorder because of reports of apparent cosegregation between this disorder and Darier's disease in a small number of families. Findings from a recent linkage study suggest that this chromosomal region may contain a susceptibility gene for bipolar affective disorder. We have found evidence of an allelic association between bipolar disorder and a marker at the pancreatic phospholipase A2 gene (PLA2A) in this region (p < or = 0.01). These results are consistent with the linkage study, and warrant additional investigations.


High-resolution crystal structure of arthropod Eiger TNF suggests a mode of receptor engagement and altered surface charge within endosomes.
The tumour necrosis factor alpha (TNFα) superfamily of proteins are critical in numerous biological processes, such as in development and immunity. Eiger is the sole TNFα member described in arthropods such as in the important model organism Drosophila. To date there are no structural data on any Eiger protein. Here we present the structure of the TNF domain of Eiger from the fall armyworm Spodoptera frugiperda (SfEiger) to 1.7 Å from a serendipitously obtained crystal without prior knowledge of the protein sequence. Our structure confirms that canonical trimerization is conserved from ancestral TNFs and points towards a mode of receptor engagement. Furthermore, we observe numerous surface histidines on SfEiger, potentially acting as pH switches following internalization into endosomes. Our data contributes to the genome annotation of S. frugiperda, a voracious agricultural pest, and can serve as a basis for future structure-function investigations of the TNF system in related arthropods such as Drosophila.


Intravenous opioids for severe acute pain in the emergency department.
To review clinical trials of intravenous opioids for severe acute pain in the emergency department (ED) and to provide an approach for optimization of therapy. Articles were identified through a search of Ovid/MEDLINE (1948-August 2010), PubMed (1950-August 2010), Cochrane Central Register of Controlled Trials (1991-August 2010), and Google Scholar (1900-August 2010). The search terms used were pain, opioid, and emergency department. The search was limited by age group to adults and by publication type to comparative studies. Studies comparing routes of administration other than intravenous or using non-opioid comparators were not included. Bibliographies of all retrieved articles were reviewed to obtain additional articles. The focus of the search was to identify original research that compared intravenous opioids used for treatment of severe acute pain for adults in the ED. At equipotent doses, randomized controlled trials have not shown clinically significant differences in analgesic response or adverse effects between opioids studied. Single opioid doses less than 0.1 mg/kg of intravenous morphine, 0.015 mg/kg of intravenous hydromorphone, or 1 μg/kg of intravenous fentanyl are likely to be inadequate for severe, acute pain and the need for additional doses should be anticipated. In none of the randomized controlled trials did patients develop respiratory depression requiring the use of naloxone. Future trials could investigate the safety and efficacy of higher doses of opioids. Implementation of nurse-initiated and patient-driven pain management protocols for opioids in the ED has shown improvements in timely provision of appropriate analgesics and has resulted in better pain reduction. Currently, intravenous administration of opioids for severe acute pain in the ED appears to be inadequate. Opioid doses in the ED should be high enough to provide adequate analgesia without additional risk to the patient. EDs could implement institution-specific protocols to standardize the management of pain.


The MOS 36-Item Short-Form Health Survey (SF-36): II. Psychometric and clinical tests of validity in measuring physical and mental health constructs.
Cross-sectional data from the Medical Outcomes Study (MOS) were analyzed to test the validity of the MOS 36-Item Short-Form Health Survey (SF-36) scales as measures of physical and mental health constructs. Results from traditional psychometric and clinical tests of validity were compared. Principal components analysis was used to test for hypothesized physical and mental health dimensions. For purposes of clinical tests of validity, clinical criteria defined mutually exclusive adult patient groups differing in severity of medical and psychiatric conditions. Scales shown in the components analysis to primarily measure physical health (physical functioning and role limitations-physical) best distinguished groups differing in severity of chronic medical condition and had the most pure physical health interpretation. Scales shown to primarily measure mental health (mental health and role limitations-emotional) best distinguished groups differing in the presence and severity of psychiatric disorders and had the most pure mental health interpretation. The social functioning, vitality, and general health perceptions scales measured both physical and mental health components and, thus, had the most complex interpretation. These results are useful in establishing guidelines for the interpretation of each scale and in documenting the size of differences between clinical groups that should be considered very large.


Genetic considerations.
Dual antiplatelet therapy with aspirin and a P2Y(12) receptor antagonist improves outcomes in patients with acute coronary syndrome and in those treated with percutaneous coronary intervention (PCI) and a coronary stent. Candidate gene and genome-wide association studies have found that common genetic polymorphisms of the cytochrome P450 (CYP) 2C19 isoenzyme that result in a loss of functional activity are associated with less exposure of clopidogrel active metabolite and a diminished antiplatelet effect. Meta-analyses of registries and genetic substudies of randomized clinical trials demonstrate that carriers of these polymorphisms who are treated with clopidogrel are at an increased risk of cardiovascular events, particularly stent thrombosis, compared with noncarriers. This deleterious effect appears to be attenuated in patients not treated with PCI. The influence of polymorphisms of other genes, such as ABCB1, is inconsistent across clinical studies. The clinical efficacy of the newer P2Y(12) antagonists prasugrel and ticagrelor do not appear to be affected by the CYP2C19 genotype, but these agents increase major bleeding not related to coronary artery bypass surgery. Although data from randomized clinical trials are currently lacking, these observations suggest that a pharmacogenomic-guided approach to antiplatelet therapy in acute coronary syndrome could potentially maximize ischemic benefit while minimizing bleeding risk.


Neural correlates of first-person perspective as one constituent of human self-consciousness.
Taking the first-person perspective (1PP) centered upon one's own body as opposed to the third-person perspective (3PP), which enables us to take the viewpoint of someone else, is constitutive for human self-consciousness. At the underlying representational or cognitive level, these operations are processed in an egocentric reference frame, where locations are represented centered around another person's (3PP) or one's own perspective (1PP). To study 3PP and 1PP, both operating in egocentric frames, a virtual scene with an avatar and red balls in a room was presented from different camera viewpoints to normal volunteers (n = 11) in a functional magnetic resonance imaging experiment. The task for the subjects was to count the objects as seen either from the avatar's perspective (3PP) or one's own perspective (1PP). The scene was presented either from a ground view (GV ) or an aerial view (AV ) to investigate the effect of view on perspective taking. The factors perspective (3PP vs. 1PP) and view (GV vs. AV ) were arranged in a two-factorial way. Reaction times were increased and percent correctness scores were decreased in 3PP as opposed to 1PP. To detect the neural mechanisms associated with perspective taking, functional magnetic resonance imaging was employed. Data were analyzed using SPM'99 in each subject and non-parametric statistics on the group level. Activations common to 3PP and 1PP (relative to baseline) were observed in a network of occipital, parietal, and prefrontal areas. Deactivations common to 3PP and 1PP (relative to baseline) were observed predominantly in mesial (i.e., parasagittal) cortical and lateral superior temporal areas bilaterally. Differential increases of neural activity were found in mesial superior parietal and right premotor cortex during 3PP (relative to 1PP), whereas differential increases during 1PP (relative to 3PP) were found in mesial prefrontal cortex, posterior cingulate cortex, and superior temporal cortex bilaterally. The data suggest that in addition to joint neural mechanisms, for example, due to visuospatial processing and decision making, 3PP and 1PP rely on differential neural processes. Mesial cortical areas are involved in decisional processes when the spatial task is solved from one's own viewpoint, whereas egocentric operations from another person's perspective differentially draw upon cortical areas known to be involved in spatial cognition.


Moving cast shadows induce apparent motion in depth.
Phenomenally strong visual illusions are described in which the motion of an object's cast shadow determines the perceived 3-D trajectory of the object. Simply adjusting the motion of a shadow is sufficient to induce dramatically different apparent trajectories of the object casting the shadow. Psychophysical results obtained with the use of 3-D graphics are reported which show that: (i) the information provided by the motion of an object's shadow overrides other strong sources of information and perceptual biases, such as the assumption of constant object size and a general viewpoint; (ii) the natural constraint of shadow darkness plays a role in the interpretation of a moving image patch as a shadow, but under some conditions even unnatural light shadows can induce apparent motion in depth of an object; (iii) when shadow motion is caused by a moving light source, the visual system incorrectly interprets the shadow motion as consistent with a moving object, rather than a moving light source. The results support the hypothesis that the human visual system incorporates a stationary light-source constraint in the perceptual processing of spatial layout of scenes.


Early maternal separation followed by later stressors leads to dysregulation of the HPA-axis and increases in hippocampal NGF and NT-3 levels in a rat model.
Early adverse life events, followed by subsequent stressors, appear to increase susceptibility for subsequent onset of psychiatric disorders in humans. The molecular mechanisms that underlie this phenomenon remain unclear, but dysregulation of the HPA axis and alterations in neurotrophic factors have been implicated. The present study investigated the effects in rodents of early maternal separation, followed by stress in adolescence and adulthood on later HPA-axis activity and hippocampal neurotrophin levels (brain-derived neurotrophic factor, nerve growth factor, and neurotrophin-3). Animals subjected to repeated stressors showed a significant decrease in basal ACTH (p < 0.05) and CORT (p < 0.05) levels when compared to controls, as well as significantly increased levels of NGF in the dorsal (p < 0.001) and ventral hippocampus (p < 0.01), and of NT-3 in the dorsal hippocampus (p < 0.01). Dysregulation of the HPA axis after multiple stressors is consistent with previous preclinical and clinical work. Given that neurotrophins are important in neuronal survival and plasticity, it is possible to speculate that their elevation reflects a compensatory mechanism.


Host density and human activities mediate increased parasite prevalence and richness in primates threatened by habitat loss and fragmentation.
1. Habitat loss and fragmentation are the principal causes of the loss of biological diversity. In addition, parasitic diseases are an emerging threat to many animals. Nevertheless, relatively few studies have tested how habitat loss and fragmentation influence the prevalence and richness of parasites in animals. 2. Several studies of nonhuman primates have shown that measures of human activity and forest fragmentation correlate with parasitism in primates. However, these studies have not tested for the ecological mechanism(s) by which human activities or forest fragmentation influence the prevalence and richness of parasites. 3. We tested the hypothesis that increased host density due to forest fragmentation and loss mediates increases in the prevalence and richness of gastrointestinal parasites in two forest primates, the Tana River red colobus (Procolobus rufomitratus, Peters 1879) and mangabey (Cercocebus galeritus galeritus, Peters 1879). We focused on population density because epidemiological theory states that host density is a key determinant of the prevalence and richness of directly transmitted parasites in animals. 4. The Tana River red colobus and mangabey are endemic to a highly fragmented forest ecosystem in eastern Kenya where habitat changes are caused by a growing human population increasingly dependent on forest resources and on clearing forest for cultivation. 5. We found that the prevalence of parasites in the two monkeys was very high compared to primates elsewhere. Density of monkeys was positively associated with forest area and disturbance in forests. In turn, the prevalence and richness of parasites was significantly associated with monkey density, and attributes indicative of human disturbance in forests. 6. We also found significant differences in the patterns of parasitism between the colobus and the mangabey possibly attributable to differences in their behavioural ecology. Colobus are arboreal folivores while mangabeys are terrestrial habitat generalists.


Top-down proteomics reveals a unique protein S-thiolation switch in Salmonella Typhimurium in response to infection-like conditions.
Characterization of the mature protein complement in cells is crucial for a better understanding of cellular processes on a systems-wide scale. Toward this end, we used single-dimension ultra-high-pressure liquid chromatography mass spectrometry to investigate the comprehensive "intact" proteome of the Gram-negative bacterial pathogen Salmonella Typhimurium. Top-down proteomics analysis revealed 563 unique proteins including 1,665 proteoforms generated by posttranslational modifications (PTMs), representing the largest microbial top-down dataset reported to date. We confirmed many previously recognized aspects of Salmonella biology and bacterial PTMs, and our analysis also revealed several additional biological insights. Of particular interest was differential utilization of the protein S-thiolation forms S-glutathionylation and S-cysteinylation in response to infection-like conditions versus basal conditions. This finding of a S-glutathionylation-to-S-cysteinylation switch in a condition-specific manner was corroborated by bottom-up proteomics data and further by changes in corresponding biosynthetic pathways under infection-like conditions and during actual infection of host cells. This differential utilization highlights underlying metabolic mechanisms that modulate changes in cellular signaling, and represents a report of S-cysteinylation in Gram-negative bacteria. Additionally, the functional relevance of these PTMs was supported by protein structure and gene deletion analyses. The demonstrated utility of our simple proteome-wide intact protein level measurement strategy for gaining biological insight should promote broader adoption and applications of top-down proteomics approaches.


Nerve growth factor infusion in the primate brain reduces lesion-induced cholinergic neuronal degeneration.
NGF is a protein that promotes survival, differentiation, and process extension of selected neuronal populations during development and, in some cases, in the mature organism. Previous lesion and aging studies in the rat have shown that intracerebroventricular NGF infusions can prevent degenerative changes in basal forebrain cholinergic neurons. We sought to determine whether salutory effects of NGF occur in the primate brain. Cholinergic fibers of the septohippocampal projection in the primate were surgically transected, followed by infusion of either a vehicle or an NGF solution into the ventricular system for a 4-week period. Quantification of cholinergic neurons in the medial septal nucleus at the end of the infusion period demonstrated that only 45 +/- 5% of cholinergic neurons could be identified after fornix lesions in vehicle-infused animals, whereas 80 +/- 6% of neurons were visible in NGF-treated animals. Thus, NGF substantially reduced lesion-induced cholinergic neuronal degeneration in the adult primate brain. This finding may be relevant to the hypothesis that NGF has potential use as a cholinergic "neurotrophic-factor therapy," given that loss of basal forebrain cholinergic neurons is common in Alzheimer's disease.


Larynx decellularization: combining freeze-drying and sonication as an effective method.
Ideal methods for the reconstruction of the laryngeal structure and restoration of the laryngeal function once the larynx has been damaged or removed have not yet been developed. Thus, larynx tissue engineering practices have recently been extensively investigated. A scaffold may be generated using biocompatible or artificial materials. Decellularization methods, which use preexisting tissues as material sources, have also been used to manufacture larynx scaffolds with promising results. In this study, we developed a novel decellularization method that combines freezing, drying, and sonication. Porcine model study. Fresh porcine larynxes were used for decellularization. The process of the decellularization cycle comprised overnight freeze-drying, defreezing in phosphate-buffered saline (PBS) for 30 minutes, and washing in PBS for another 30 minutes. Sonication treatment was further added during the defreezing process. The decellularized tissue was then evaluated through histologic sections under hematoxylin and eosin staining. The results showed that a single use of the freeze-drying modality has little effect with regards to removing cellular components, even with increased decellularization cycles. However, when sonication was added to the defreezing process, the cellular contents were removed significantly (the residual nucleus ratios of freeze-drying:freeze-drying and defreezing one cycle:freeze-drying and defreezing three cycles:freeze-drying and defreezing under sonication three cycles were 91%:70%:47%:16%, respectively). However, the processed scaffold became structurally more fragile through the procedure. Combining freeze-drying and sonication during the defreezing process could be a promising method of decellularizing laryngeal tissues. However, this purely physical method may also induce structural damage to the scaffold.


Interacting effects of phenotypic plasticity and evolution on population persistence in a changing climate.
Climate change affects individual organisms by altering development, physiology, behavior, and fitness, and populations by altering genetic and phenotypic composition, vital rates, and dynamics. We sought to clarify how selection, phenotypic plasticity, and demography are linked in the context of climate change. On the basis of theory and results of recent empirical studies of plants and animals, we believe the ecological and evolutionary issues relevant to population persistence as climate changes are the rate, type, magnitude, and spatial pattern of climate-induced abiotic and biotic change; generation time and life history of the organism; extent and type of phenotypic plasticity; amount and distribution of adaptive genetic variation across space and time; dispersal potential; and size and connectivity of subpopulations. An understanding of limits to plasticity and evolutionary potential across traits, populations, and species and feedbacks between adaptive and demographic responses is lacking. Integrated knowledge of coupled ecological and evolutionary mechanisms will increase understanding of the resilience and probabilities of persistence of populations and species.


Bidirectional promoters generate pervasive transcription in yeast.
Genome-wide pervasive transcription has been reported in many eukaryotic organisms, revealing a highly interleaved transcriptome organization that involves hundreds of previously unknown non-coding RNAs. These recently identified transcripts either exist stably in cells (stable unannotated transcripts, SUTs) or are rapidly degraded by the RNA surveillance pathway (cryptic unstable transcripts, CUTs). One characteristic of pervasive transcription is the extensive overlap of SUTs and CUTs with previously annotated features, which prompts questions regarding how these transcripts are generated, and whether they exert function. Single-gene studies have shown that transcription of SUTs and CUTs can be functional, through mechanisms involving the generated RNAs or their generation itself. So far, a complete transcriptome architecture including SUTs and CUTs has not been described in any organism. Knowledge about the position and genome-wide arrangement of these transcripts will be instrumental in understanding their function. Here we provide a comprehensive analysis of these transcripts in the context of multiple conditions, a mutant of the exosome machinery and different strain backgrounds of Saccharomyces cerevisiae. We show that both SUTs and CUTs display distinct patterns of distribution at specific locations. Most of the newly identified transcripts initiate from nucleosome-free regions (NFRs) associated with the promoters of other transcripts (mostly protein-coding genes), or from NFRs at the 3' ends of protein-coding genes. Likewise, about half of all coding transcripts initiate from NFRs associated with promoters of other transcripts. These data change our view of how a genome is transcribed, indicating that bidirectionality is an inherent feature of promoters. Such an arrangement of divergent and overlapping transcripts may provide a mechanism for local spreading of regulatory signals-that is, coupling the transcriptional regulation of neighbouring genes by means of transcriptional interference or histone modification.


Am I too fat to be a princess? Examining the effects of popular children's media on young girls' body image.
The current study investigated the effects of brief exposure to appearance-related media on young girls' body image. One hundred and twenty-one girls aged 3-6 years old participated. Results indicated that exposure did not affect body dissatisfaction or engagement in appearance-related play behaviours. This is the first empirical study to provide support for previous findings that suggest media exposure does not affect body image in young girls. In contrast to older populations, it is possible that young children may adopt the persona of attractive characters with whom they identify rather than comparing themselves to the characters. Although nearly all girls liked the way they looked, self-report data indicated that nearly one-third of the participants would change something about their physical appearance and nearly half of the girls worried about being fat. Exposure to appearance-related media did not exacerbate concerns.


Semaphorin 3A is a new early diagnostic biomarker of experimental and pediatric acute kidney injury.
Semaphorin 3A is a secreted protein that regulates cell motility and attachment in axon guidance, vascular growth, immune cell regulation and tumor progression. However, nothing is known about its role in kidney pathophysiology. Here, we determined whether semaphorin3A is induced after acute kidney injury (AKI) and whether urinary semaphorin 3A can predict AKI in humans undergoing cardiopulmonary bypass (CPB). In animals, semaphorin 3A is localized in distal tubules of the kidney and excretion increased within 3 hr after reperfusion of the kidney whereas serum creatinine was significantly raised at 24 hr. In humans, using serum creatinine, AKI was detected on average only 48 hours after CPB. In contrast, urine semaphorin increased at 2 hours after CPB, peaked at 6 hours (2596±591 pg/mg creatinine), and was no longer significantly elevated 12 hours after CPB. The predictive power of semaphorin 3A as demonstrated by area under the receiver-operating characteristic curve for diagnosis of AKI at 2, 6, and 12 hours after CPB was 0.88, 0.81, and 0.74, respectively. The 2-hour urine semaphorin measurement strongly correlated with duration and severity of AKI, as well as length of hospital stay. Adjusting for CPB time and gender, the 2-hour semaphorin remained an independent predictor of AKI, with an odds ratio of 2.19. Our results suggest that semaphorin 3A is an early, predictive biomarker in experimental and pediatric AKI, and may allow for the reliable early diagnosis and prognosis of AKI after CPB, much before the rise in serum creatinine.


Oxidative stress: an essential factor in the pathogenesis of gastrointestinal mucosal diseases.
Reactive oxygen species (ROS) are generated as by-products of normal cellular metabolic activities. Superoxide dismutase, glutathione peroxidase, and catalase are the enzymes involved in protecting cells from the damaging effects of ROS. ROS are produced in response to ultraviolet radiation, cigarette smoking, alcohol, nonsteroidal anti-inflammatory drugs, ischemia-reperfusion injury, chronic infections, and inflammatory disorders. Disruption of normal cellular homeostasis by redox signaling may result in cardiovascular, neurodegenerative diseases and cancer. ROS are produced within the gastrointestinal (GI) tract, but their roles in pathophysiology and disease pathogenesis have not been well studied. Despite the protective barrier provided by the mucosa, ingested materials and microbial pathogens can induce oxidative injury and GI inflammatory responses involving the epithelium and immune/inflammatory cells. The pathogenesis of various GI diseases including peptic ulcers, gastrointestinal cancers, and inflammatory bowel disease is in part due to oxidative stress. Unraveling the signaling events initiated at the cellular level by oxidative free radicals as well as the physiological responses to such stress is important to better understand disease pathogenesis and to develop new therapies to manage a variety of conditions for which current therapies are not always sufficient.


An Integrative Approach Uncovers Biomarkers that Associate with Clinically Relevant Disease Outcomes in Vulvar Carcinoma.
Vulvar squamous cell carcinoma (VSCC) is a rare disease that has a high mortality rate (∼40%). However, little is known about its molecular signature. Therefore, an integrated genomics approach, based on comparative genome hybridization (aCGH) and genome-wide expression (GWE) array, was performed to identify driver genes in VSCC. To achieve that, DNA and RNA were extracted from frozen VSCC clinical specimens and examined by aCGH and GWE array, respectively. On the basis of the integration of data using the CONEXIC algorithm, PLXDC2 and GNB3 were validated by RT-qPCR. The expression of these genes was then analyzed by IHC in a large set of formalin-fixed paraffin-embedded specimens. These analyses identified 47 putative drivers, 46 of which were characterized by copy number gains that were concomitant with overexpression and one with a copy number loss and downregulation. Two of these genes, PLXDC2 and GNB3, were selected for further validation: PLXDC2 was downregulated and GNB3 was overexpressed compared with non-neoplastic tissue. By IHC, both proteins were ubiquitously expressed throughout vulvar tissue. High expression of GNB3 and low PLXDC2 immunostaining in the same sample was significantly associated with less lymph node metastasis and greater disease-free survival. On the basis of a robust methodology never used before for VSCC evaluation, two novel prognostic markers in vulvar cancer are identified: one with favorable prognosis (GNB3) and the other with unfavorable prognosis (PLXDC2). This genomics study reveals markers that associate with prognosis and may provide guidance for better treatment in vulvar cancer. Mol Cancer Res; 14(8); 720-9. ©2016 AACR.


What Is the Survivorship of Revision Surgery Performed for the Chronically Dislocated THA?
Instability remains one of the most common indications for revision THA. However, little is known about the efficacy of surgery for and the complications associated with revision THA for patients with a chronically dislocated THA, which we define as a dislocation of more than 4 weeks. For patients with a chronically dislocated THA undergoing revision THA, we asked (1) What is the survivorship free from additional revision for these procedures? (2) What complications are associated with revision THA in this setting? (3) What are the clinical outcomes as measured by the Harris hip score in these procedures? From 1998 to 2014, 1084 patients who underwent revision THA for instability were reviewed and 33 patients (33 hips) were identified who had a hip that had been dislocated for more than 4 weeks. Median time dislocated was 4 months (range, 1-120 months), and the mean distance of the femoral head above hip center at presentation was 45 mm. Mean patient age was 67 ± 17 years, and 79% of patients (26 of 33) were women. During the period in question, we used four approaches: Treatment with acetabular component revision in 18 of 33 patients (55%), head and liner exchange in nine patients (27%), both-component revision in five patients (15%), and isolated femoral component revision in one patient (3%). A constrained liner was used in 17 patients (52%), including six of the patients treated with acetabular component revision, and three of those who had both-component revisions. During the period in question, our general indications were hip pain and/or unacceptable function with the chronically dislocated prosthesis. Our sample size was too small to evaluate the association of the procedure choice on survivorship or complication risk. We used Kaplan-Meier survivorship analysis to estimate survivorship free from complication, reoperation, or revision. Mean followup was 4.4 years (range, 2-10 years). Survivorship free from any revision, complication, or reoperation was 61% at 5 years (95% CI, 43-82). Survivorship free from revision was 83% at 5 years (95% CI, 67-98). Etiology for revision was aseptic loosening in three of 33 hips (9%), recurrent dislocation in two hips (6%), and deep periprosthetic joint infection in two hips (6%). Five complications (15%) did not result in a reoperation, including one dislocation and one incomplete peroneal nerve palsy in a patient after an anterolateral approach. The Harris hip score improved from mean 50 ± 17 preoperatively to mean 80 ± 11 at 5 years. Chronically dislocated THAs can be successfully managed with revision THA. We recommend close evaluation of the components for aseptic loosening, performing revision surgery only on patients with pain and poor function, and thoroughly counseling patients that survivorship is modest and complications are common. Level IV, therapeutic study.


Novel MR imaging method--MAVRIC--for metal artifact suppression after joint replacement in musculoskeletal tumor patients.
Standard imaging modality for the follow-up after prosthetic replacements for musculoskeletal tumor patients has been conventional radiography. This technique is effective in detecting subtle changes in bone adjacent to metal implants, but in many cases, radiographs do not lead to definitive diagnosis of postoperative adverse events such as acute infection, local recurrence of soft tissue tumor or soft tissue local recurrence of osseous sarcoma. Conventional MRI sequences have not been effective due to metal artifacts. In this study, we tried to elucidate the effectiveness of metal artifact suppression using novel sequence, multiacquisition variable-resonance image combination (MAVRIC), after musculoskeletal tumor surgeries. We retrospectively analyzed 5 cases of malignant bone and soft tissue sarcoma patients who were reconstructed with metal prosthesis after wide resection of tumors. Images obtained using MAVRIC and short tau inversion recovery (STIR) were compared side by side. The paired MAVRIC and STIR images were qualitatively compared independently by two specialists for 4 parameters: visualization of bone - implant interface, visualization of surrounding soft tissues, image blurring, and overall image quality. Quantitatively, paired images were reviewed to identify the slice where the metal artifact was maximal, and a region of interest encompassing the implant and surrounding artifact was drawn using Advantage Workstation (GE Healthcare, Japan). There were no local recurrences that were detected. By utilizing MAVRIC, visualization of the bone - implant interface and visualization of the surrounding soft tissue were significantly improved in MAVRIC compared to STIR. Although blurring was worse on the MAVRIC acquisitions, the overall image quality was still better on MAVRIC. Quantitatively, the area of metal artifact measured using MAVRIC was markedly less compared to STIR (61.4 cm(2) vs 135.9 cm(2)). Despite the relatively small number of cases in the present study, our observation strongly suggests that MAVRIC is able to improve the quality of images by decreasing the artifact caused by endoprosthesis, frequently utilized in reconstruction of musculoskeletal tumor patients. Further installments of conventional imaging sequences with the addition of gadolinium - enhancement will enable increased accuracy in diagnosing local recurrences of sarcoma patients.