Shame! Self-stigmatisation as an obstacle to sick doctors returning to work: a qualitative study.
To explore the views of sick doctors on the obstacles preventing them returning to work. Qualitative study. Single participating centre recruiting doctors from all over the UK. Doctors who had been away from work for at least 6 months with physical or mental health problems, drug or alcohol problems, General Medical Council involvement or any combination of these, were eligible. Eligible doctors were recruited in conjunction with the Royal Medical Benevolent Fund, the General Medical Council and the Practitioner Health Programme. These organisations approached 77 doctors; 19 participated. Each doctor completed an in-depth semistructured interview. We used a constant comparison method to identify and agree on the coding of the data and the identification of a number of central themes. The doctors described that being away from work left them isolated and sad. Many experienced negative reactions from their family and some deliberately concealed their problems. Doctors described a lack of support from colleagues and feared a negative response when returning to work. Self-stigmatisation was central to the participants' accounts; several described themselves as failures and appeared to have internalised the negative views of others. Self-stigmatising views, which possibly emerge from the belief that 'doctors are invincible', represent a major obstacle to doctors returning to work. From medical school onwards cultural change is necessary to allow doctors to recognise their vulnerabilities so they can more easily generate strategies to manage if they become unwell.


Haemodynamic responses to static and dynamic handgrip before and after autonomic blockade.
1. Six healthy men performed static and dynamic handgrip to local muscular fatigue in approximately 6 min under control conditions, i.e. without drugs and after combined parasympathetic and beta-adrenergic blockade with atropine and metoprolol. 2. From rest to exercise at fatigue, systolic, diastolic and mean arterial pressures increased by 32 +/- 4 and 39 +/- 3 mmHg, 24 +/- 3 and 26 +/- 4 mmHg, and 26 +/- 3 and 30 +/- 3 mmHg respectively for static and dynamic handgrip. There were no significant differences between the pressor responses for the two modes of contraction. Cardiac output increased significantly only during dynamic exercise. Total peripheral resistance increased by 2.3 +/- 1.0 units for static handgrip (P less than 0.05) and by 0.7 +/- 0.8 unit (P greater than 0.05) for dynamic handgrip. Autonomic blockade abolished the heart rate response to both static and dynamic handgrip. For both modes of contraction the systolic arterial pressure responses were 9-12 mmHg lower (P less than 0.05) after autonomic blockade, but the diastolic and mean pressure responses were not significantly affected. A significant increase in cardiac output persisted during dynamic exercise. The increase in peripheral resistance during static exercise tended to be greater after blockade. Plasma noradrenaline and adrenaline levels showed only minor elevations in response to static and dynamic handgrip and were not changed by autonomic blockade. 3. These data indicate that when performed to a common end-point with identical small muscle groups static and dynamic exercise produce an equally large pressor response, which is only slightly attenuated by autonomic blockade.


Dual cholinergic signals regulate daily migration of hematopoietic stem cells and leukocytes.
Hematopoietic stem and progenitor cells (HSPCs) and leukocytes circulate between the bone marrow (BM) and peripheral blood following circadian oscillations. Autonomic sympathetic noradrenergic signals have been shown to regulate HSPC and leukocyte trafficking, but the role of the cholinergic branch has remained unexplored. We have investigated the role of the cholinergic nervous system in the regulation of day/night traffic of HSPCs and leukocytes in mice. We show here that the autonomic cholinergic nervous system (including parasympathetic and sympathetic) dually regulates daily migration of HSPCs and leukocytes. At night, central parasympathetic cholinergic signals dampen sympathetic noradrenergic tone and decrease BM egress of HSPCs and leukocytes. However, during the daytime, derepressed sympathetic noradrenergic activity causes predominant BM egress of HSPCs and leukocytes via β3-adrenergic receptor. This egress is locally supported by light-triggered sympathetic cholinergic activity, which inhibits BM vascular cell adhesion and homing. In summary, central (parasympathetic) and local (sympathetic) cholinergic signals regulate day/night oscillations of circulating HSPCs and leukocytes. This study shows how both branches of the autonomic nervous system cooperate to orchestrate daily traffic of HSPCs and leukocytes.


Data collected by coupling fix and wearable sensors for addressing urban microclimate variability in an historical Italian city.
This article presents the data collected through an extensive research work conducted in a historic hilly town in central Italy during the period 2016-2017. Data concern two different datasets: long-term hygrothermal histories collected in two specific positions of the town object of the research, and three environmental transects collected following on foot the same designed path at three different time of the same day, i.e. during a heat wave event in summer. The short-term monitoring campaign is carried out by means of an innovative wearable weather station specifically developed by the authors and settled upon a bike helmet. Data provided within the short-term monitoring campaign are analysed by computing the apparent temperature, a direct indicator of human thermal comfort in the outdoors. All provided environmental data are geo-referenced. These data are used in order to examine the intra-urban microclimate variability. Outcomes from both long- and short-term monitoring campaigns allow to confirm the existing correlation between the urban forms and functionalities and the corresponding local microclimate conditions, also generated by anthropogenic actions. In detail, higher fractions of built surfaces are associated to generally higher temperatures as emerges by comparing the two long-term air temperature data series, i.e. temperature collected at point 1 is higher than temperature collated at point 2 for the 75% of the monitored period with an average of +2.8 °C. Furthermore, gathered environmental transects demonstrate the high variability of the main environmental parameters below the Urban Canopy. Diversification of the urban thermal behaviour leads to a computed apparent temperature range in between 33.2 °C and 46.7 °C at 2 p.m. along the monitoring path. Reuse of these data may be helpful for further investigating interesting correlations among urban configuration, anthropogenic actions and microclimate variables affecting outdoor comfort. Additionally, the proposed dataset may be compared to other similar datasets collected in other urban contexts around the world. Finally, it can be compared to other monitoring methodologies such as weather stations and satellite measurements available in the location at the same time.


Miniaturised nucleic acid analysis.
The application of micro total analysis systems has grown exponentially over the past few years, particularly diversifying in disciplines related to bioassays. The primary focus of this review is to detail recent new approaches to sample preparation, nucleic acid amplification and detection within microfluidic devices or at the microscale level. We also introduce some applications that have as yet to be explored in a miniaturised environment, but should benefit from improvements in analytical efficiency and functionality when transferred to planar-chip formats. The studies described in this review were published in commonly available journals as well as in the proceedings of three major conferences relevant to microfluidics (Micro Total Analysis Systems, Transducers and The Nanotechnology Conference and Trade Show). Although an emphasis has been placed on papers published since 2002, pertinent articles preceding this publication year have also been included.


Carbohydrates as additives for the formation of isoporous PS-b-P4VP diblock copolymer membranes.
Highly porous polystyrene-block-poly(4-vinylpyridine) (PS-b-P4VP) diblock copolymer membranes are prepared using carbohydrates as additives. Therefore α-cyclodextrine, α-(D)-glucose, and saccharose (cane sugar) are tested for the membrane formation of three different PS-b-P4VP polymers. The addition of the carbohydrates leads to an increasing viscosity of the membrane solutions due to hydrogen bonding between hydroxyl groups of the carbohydrates and pyridine units of the block copolymer. In all cases, the membranes made from solution with carbohydrates have higher porosity, an improved narrow pore distribution on the surface and a higher water flux as membranes made without carbohydrates with the same polymer, solvent ratio, and polymer concentration.


Evaluation of premature senescence and senescence biomarkers in carcinoma cells and xenograft mice exposed to single or fractionated irradiation.
The purpose of the present study was to elucidate whether premature senescence contributes to the outcome of radiotherapy (RT) and to validate senescence biomarkers in vitro and in vivo. Cultured human cancer cell lines and xenografted mice were exposed to single (SR; 2, 6 or 12 Gy) or fractionated radiation (FR; 3 x 2 Gy or 6 x 2 Gy), and premature senescence was assessed using senescence-associated β-galactosidase (SA-β-Gal) activity, hypophosphorylation of pRb and p21 accumulation. A variety of senescence-associated biomarkers including cathepsin D (CD), the eukaryotic translation elongation factors eEF1A1, eEF1B2, decoy receptor 2 and Dec1 were further validated in vivo or in vitro. We demonstrated the beneficial tumor suppressive role of ionizing radiation (IR)-induced premature senescence in vitro and in vivo. FR inhibited tumor growth via induction of premature senescence as effectively as an equivalent SR dose (≥6 Gy). In addition, CD and eEF1 were valuable biomarkers of cellular senescence in either SR- or RF-exposed carcinoma cells or xenograft mice. Our results suggest that 2 Gy of a conventional RT regime could achieve a better clinical outcome if premature senescence could be increased through an improved understanding of its molecular action mechanism.


Alpha lipoic acid inhibits oxidative stress-induced apoptosis by modulating of Nrf2 signalling pathway after traumatic brain injury.
Alpha lipoic acid (ALA) is a powerful antioxidant which has been widely used in the treatment of different system diseases, such as cardiovascular and cerebrovascular diseases. But, there are few studies that refer to protective effects and potential mechanisms on traumatic brain injury (TBI). This study was carried out to investigate the neuroprotective effect following TBI and illuminate the underlying mechanism. Weight drop-injured model in rats was induced by weight-drop. ALA was administrated via intraperitoneal injection after TBI. Neurologic scores were examined following several tests. Neurological score was performed to measure behavioural outcomes. Nissl staining and TUNEL were performed to evaluate the neuronal apoptosis. Western blotting was engaged to analyse the protein content of the Nuclear factor erythroid 2-related factor 2 (Nrf2) and its downstream protein factors, including hemeoxygenase-1 (HO-1) and quinine oxidoreductase-1 (NQO1). ALA treatment alleviated TBI-induced neuron cell apoptosis and improved neurobehavioural function by up-regulation of Nrf2 expression and its downstream protein factors after TBI. This study presents new perspective of the mechanisms responsible for the neuronal apoptosis of ALA, with possible involvement of Nrf2 pathway.


The public costs of births resulting from unintended pregnancies: national and state-level estimates.
Births resulting from unintended pregnancies are associated with substantial maternity and infant care costs to the federal and state governments; these costs have never been estimated at the national and state levels. The proportions of births paid for by public insurance programs in 2006 were estimated, by pregnancy intention status, using data from the Pregnancy Risk Assessment Monitoring System and similar state surveys, or were predicted by multivariate linear regression. Public costs were calculated using state-level estimates of the number of births, by intention status, and of the cost of a publicly funded birth. In 2006, 64% of births resulting from unintended pregnancies were publicly funded, compared with 48% of all births and 35% of births resulting from intended pregnancies. The proportion of births resulting from unintended pregnancies that were publicly funded varied by state, from 42% to 81%. Of the 2.0 million publicly funded births, 51% resulted from unintended pregnancies, accounting for $11.1 billion in costs-half of the total public expenditures on births. In seven states, the costs for births from unintended pregnancies exceeded a half billion dollars. Public insurance programs are central in assisting American families in affording pregnancy and childbirth; however, they pay for a disproportionately high number of births resulting from unintended pregnancy. The resulting budgetary impact warrants increased public efforts to reduce unintended pregnancy.


The role of the immune system in governing host-microbe interactions in the intestine.
The mammalian intestinal tract harbors a diverse community of trillions of microorganisms, which have co-evolved with the host immune system for millions of years. Many of these microorganisms perform functions critical for host physiology, but the host must remain vigilant to control the microbial community so that the symbiotic nature of the relationship is maintained. To facilitate homeostasis, the immune system ensures that the diverse microbial load is tolerated and anatomically contained, while remaining responsive to microbial breaches and invasion. Although the microbiota is required for intestinal immune development, immune responses also regulate the structure and composition of the intestinal microbiota. Here we discuss recent advances in our understanding of these complex interactions and their implications for human health and disease.


Srrm234, but not canonical SR and hnRNP proteins, drive inclusion of Dscam exon 9 variable exons.
Alternative splicing of pre-mRNA is a major mechanism to diversify protein functionality in metazoans from a limited number of genes. The Drosophila melanogaster Down syndrome cell adhesion molecule (Dscam) gene, which is important for neuronal wiring and phagocytosis of bacteria, can generate up to 38,016 isoforms by mutually exclusive alternative splicing in four clusters of variable exons. However, it is not understood how a specific exon is chosen from the many variables and how variable exons are prevented from being spliced together. A main role in the regulation of Dscam alternative splicing has been attributed to RNA binding proteins (RBPs), but how they impact on exon selection is not well understood. Serine-arginine rich (SR) proteins and hnRNP proteins are the two main types of RBPs with major roles in exon definition and splice site selection. Here, we analyzed the role of SR and hnRNP proteins in Dscam exon 9 alternative splicing in mutant Drosophila melanogaster embryos because of their essential function for development. Strikingly, loss or overexpression of canonical SR and hnRNP proteins even when multiple proteins are depleted together, does not affect Dscam alternative exon selection very dramatically. Conversely, noncanonical SR protein Serine-arginine repetitive matrix 2/3/4 (Srrm234) is a main determinant of exon inclusion in the Dscam exon 9 cluster. Since long-range base-pairings are absent in the exon 9 cluster, our data argue for a small complement of regulatory factors as main determinants of exon inclusion in the Dscam exon 9 cluster.


Activation of Heschl's gyrus during auditory hallucinations.
Apart from being a common feature of mental illness, auditory hallucinations provide an intriguing model for the study of internally generated sensory perceptions that are attributed to external sources. Until now, the knowledge about the cortical network that supports such hallucinations has been restricted by methodological limitations. Here, we describe an experiment with paranoid schizophrenic patients whose on- and offset of auditory hallucinations could be monitored within one functional magnetic resonance imaging (fMRI) session. We demonstrate an increase of the blood oxygen level-dependent (BOLD) signal in Heschl's gyrus during the patients' hallucinations. Our results provide direct evidence of the involvement of primary auditory areas in auditory verbal hallucinations and establish novel constraints for psychopathological models.


The histone demethylase Jarid1b mediates angiotensin II-induced endothelial dysfunction by controlling the 3'UTR of soluble epoxide hydrolase.
The histone demethylase Jarid1b limits gene expression by removing the active methyl mark from histone3 lysine4 at gene promoter regions. A vascular function of Jarid1b is unknown, but a vasoprotective function to inflammatory and hypertrophic stimuli, like angiotensin II (AngII) could be inferred. This hypothesis was tested using Jarid1b knockout mice and the inhibitor PBIT. Mice or aortic segments were treated with AngII to induce endothelial dysfunction. Aortae from WT and Jarid1b knockout were studied in organ chambers and endothelium-dependent dilator responses to acetylcholine and endothelium-independent responses to DetaNONOate were recorded after pre-constriction with phenylephrine in the presence or absence of the NO-synthase inhibitor nitro-L-arginine. Molecular mechanisms were investigated with chromatin immunoprecipitation, RNA-Seq, RNA-3'-adaptor-ligation, actinomycin D and RNA-immunoprecipitation. Knockout or inhibition of Jarid1b prevented the development of endothelial dysfunction in response to AngII. This effect was not a consequence of altered nitrite oxide availability but accompanied by a loss of the inflammatory response to AngII. As Jarid1b mainly inhibits gene expression, an indirect effect should account for this observation. AngII induced the soluble epoxide hydrolase (sEH), which degrades anti-inflammatory lipids, and thus promotes inflammation. Knockout or inhibition of Jarid1b prevented the AngII-mediated sEH induction. Mechanistically, Jarid1b maintained the length of the 3'untranslated region of the sEH mRNA, thereby increasing its stability and thus sEH protein expression. Loss of Jarid1b activity therefore resulted in sEH mRNA destabilization. Jarid1b contributes to the pro-inflammatory effects of AngII by stabilizing sEH expression. Jarid1b inhibition might be an option for future therapeutics against cardiovascular dysfunction.


Functional metagenomic profiling of nine biomes.
Microbial activities shape the biogeochemistry of the planet and macroorganism health. Determining the metabolic processes performed by microbes is important both for understanding and for manipulating ecosystems (for example, disruption of key processes that lead to disease, conservation of environmental services, and so on). Describing microbial function is hampered by the inability to culture most microbes and by high levels of genomic plasticity. Metagenomic approaches analyse microbial communities to determine the metabolic processes that are important for growth and survival in any given environment. Here we conduct a metagenomic comparison of almost 15 million sequences from 45 distinct microbiomes and, for the first time, 42 distinct viromes and show that there are strongly discriminatory metabolic profiles across environments. Most of the functional diversity was maintained in all of the communities, but the relative occurrence of metabolisms varied, and the differences between metagenomes predicted the biogeochemical conditions of each environment. The magnitude of the microbial metabolic capabilities encoded by the viromes was extensive, suggesting that they serve as a repository for storing and sharing genes among their microbial hosts and influence global evolutionary and metabolic processes.


Nodus migrans: the case of the migrating knot.
Flow-directed pulmonary artery catheters provide important information regarding intravascular volume status, cardiac function and vascular resistance. We describe an unusual complication of pulmonary artery catheterization in which a knot formed at the distal end was torn away from the catheter body and migrated from its original position in the right subclavian vein to a distal branch of the right pulmonary artery. Careful attention to insertion and withdrawal techniques could prevent this potentially serious complication.


A reconstruction of regional and global temperature for the past 11,300 years.
Surface temperature reconstructions of the past 1500 years suggest that recent warming is unprecedented in that time. Here we provide a broader perspective by reconstructing regional and global temperature anomalies for the past 11,300 years from 73 globally distributed records. Early Holocene (10,000 to 5000 years ago) warmth is followed by ~0.7°C cooling through the middle to late Holocene (<5000 years ago), culminating in the coolest temperatures of the Holocene during the Little Ice Age, about 200 years ago. This cooling is largely associated with ~2°C change in the North Atlantic. Current global temperatures of the past decade have not yet exceeded peak interglacial values but are warmer than during ~75% of the Holocene temperature history. Intergovernmental Panel on Climate Change model projections for 2100 exceed the full distribution of Holocene temperature under all plausible greenhouse gas emission scenarios.


Hodgkin Lymphoma Version 1.2017, NCCN Clinical Practice Guidelines in Oncology.
This portion of the NCCN Guidelines for Hodgkin lymphoma (HL) focuses on the management of classical HL. Current management of classical HL involves initial treatment with chemotherapy or combined modality therapy followed by restaging with PET/CT to assess treatment response using the Deauville criteria (5-point scale). The introduction of less toxic and more effective regimens has significantly advanced HL cure rates. However, long-term follow-up after completion of treatment is essential to determine potential long-term effects.


Evolutionary history and functional implications of protein domains and their combinations in eukaryotes.
In higher multicellular eukaryotes, complex protein domain combinations contribute to various cellular functions such as regulation of intercellular or intracellular signaling and interactions. To elucidate the characteristics and evolutionary mechanisms that underlie such domain combinations, it is essential to examine the different types of domains and their combinations among different groups of eukaryotes. We observed a large number of group-specific domain combinations in animals, especially in vertebrates. Examples include animal-specific combinations in tyrosine phosphorylation systems and vertebrate-specific combinations in complement and coagulation cascades. These systems apparently underwent extensive evolution in the ancestors of these groups. In extant animals, especially in vertebrates, animal-specific domains have greater connectivity than do other domains on average, and contribute to the varying number of combinations in each animal subgroup. In other groups, the connectivities of older domains were greater on average. To observe the global behavior of domain combinations during evolution, we traced the changes in domain combinations among animals and fungi in a network analysis. Our results indicate that there is a correlation between the differences in domain combinations among different phylogenetic groups and different global behaviors. Rapid emergence of animal-specific domains was observed in animals, contributing to specific domain combinations and functional diversification, but no such trends were observed in other clades of eukaryotes. We therefore suggest that the strategy for achieving complex multicellular systems in animals differs from that of other eukaryotes.


Mapping the Oxford Shoulder Score onto the EQ-5D utility index.
In order to enable cost-utility analysis of shoulder pain conditions and treatments, this study aimed to develop and evaluate mapping algorithms to estimate the EQ-5D health index from the Oxford Shoulder Score (OSS) when health outcomes are only assessed with the OSS. 5437 paired OSS and EQ-5D questionnaire responses from four national multicentre randomised controlled trials investigating different shoulder pathologies and treatments were split into training and testing samples. Separate EQ-5D-3L and EQ-5D-5L analyses were undertaken. Transfer to utility (TTU) regression (univariate linear, polynomial, spline, multivariable linear, two-part logistic-linear, tobit and adjusted limited dependent variable mixture models) and response mapping (ordered logistic regression and seemingly unrelated regression (SUR)) models were developed on the training sample. These were internally validated, and their performance evaluated on the testing sample. Model performance was evaluated over 100-fold repeated training-testing sample splits. For the EQ-5D-3L analysis, the multivariable linear and splines models had the lowest mean square error (MSE) of 0.0415. The SUR model had the lowest mean absolute error (MAE) of 0.136. Model performance was greatest in the mid-range and best health states, and lowest in poor health states. For the EQ-5D-5L analyses, the multivariable linear and splines models had the lowest MSE (0.0241-0.0278) while the SUR models had the lowest MAE (0.105-0.113). The developed models now allow accurate estimation of the EQ-5D health index when only the OSS responses are available as a measure of patient-reported health outcome.


Sustained Na+/H+ exchanger activation promotes gliotransmitter release from reactive hippocampal astrocytes following oxygen-glucose deprivation.
Hypoxia ischemia (HI)-related brain injury is the major cause of long-term morbidity in neonates. One characteristic hallmark of neonatal HI is the development of reactive astrogliosis in the hippocampus. However, the impact of reactive astrogliosis in hippocampal damage after neonatal HI is not fully understood. In the current study, we investigated the role of Na(+)/H(+) exchanger isoform 1 (NHE1) protein in mouse reactive hippocampal astrocyte function in an in vitro ischemia model (oxygen/glucose deprivation and reoxygenation, OGD/REOX). 2 h OGD significantly increased NHE1 protein expression and NHE1-mediated H(+) efflux in hippocampal astrocytes. NHE1 activity remained stimulated during 1-5 h REOX and returned to the basal level at 24 h REOX. NHE1 activation in hippocampal astrocytes resulted in intracellular Na(+) and Ca(2+) overload. The latter was mediated by reversal of Na(+)/Ca(2+) exchange. Hippocampal astrocytes also exhibited a robust release of gliotransmitters (glutamate and pro-inflammatory cytokines IL-6 and TNFα) during 1-24 h REOX. Interestingly, inhibition of NHE1 activity with its potent inhibitor HOE 642 not only reduced Na(+) overload but also gliotransmitter release from hippocampal astrocytes. The noncompetitive excitatory amino acid transporter inhibitor TBOA showed a similar effect on blocking the glutamate release. Taken together, we concluded that NHE1 plays an essential role in maintaining H(+) homeostasis in hippocampal astrocytes. Over-stimulation of NHE1 activity following in vitro ischemia disrupts Na(+) and Ca(2+) homeostasis, which reduces Na(+)-dependent glutamate uptake and promotes release of glutamate and cytokines from reactive astrocytes. Therefore, blocking sustained NHE1 activation in reactive astrocytes may provide neuroprotection following HI.


Essential molecular determinants for thyroid hormone transport and first structural implications for monocarboxylate transporter 8.
Monocarboxylate transporter 8 (MCT8, SLC16A2) is a thyroid hormone (TH) transmembrane transport protein mutated in Allan-Herndon-Dudley syndrome, a severe X-linked psychomotor retardation. The neurological and endocrine phenotypes of patients deficient in MCT8 function underscore the physiological significance of carrier-mediated TH transmembrane transport. MCT8 belongs to the major facilitator superfamily of 12 transmembrane-spanning proteins and mediates energy-independent bidirectional transport of iodothyronines across the plasma membrane. Structural information is lacking for all TH transmembrane transporters. To gain insight into structure-function relations in TH transport, we chose human MCT8 as a paradigm. We systematically performed conventional and liquid chromatography-tandem mass spectrometry-based uptake measurements into MCT8-transfected cells using a large number of compounds structurally related to iodothyronines. We found that human MCT8 is specific for L-iodothyronines and requires at least one iodine atom per aromatic ring. Neither thyronamines, decarboxylated metabolites of iodothyronines, nor triiodothyroacetic acid and tetraiodothyroacetic acid, TH derivatives lacking both chiral center and amino group, are substrates for MCT8. The polyphenolic flavonoids naringenin and F21388, potent competitors for TH binding at transthyretin, did not inhibit T(3) transport, suggesting that MCT8 can discriminate its ligand better than transthyretin. Bioinformatic studies and a first molecular homology model of MCT8 suggested amino acids potentially involved in substrate interaction. Indeed, alanine mutation of either Arg(445) (helix 8) or Asp(498) (helix 10) abrogated T(3) transport activity of MCT8, supporting their predicted role in substrate recognition. The MCT8 model allows us to rationalize potential interactions of amino acids including those mutated in patients with Allan-Herndon-Dudley syndrome.


Brillouin induced self-heterodyne technique for narrow line width measurement.
A novel technique is introduced for jitter-insensitive sub-KHz resolution linewidth characterization technique in ultra-narrow lasers for optical communication applications. The technique is based on self-heterodyne detection induced by Stimulated Brillouin Scattering (SBS). Non linear SBS drives the heterodyne mixing through optical frequency locking of a narrow tunable laser source and the signal under test, which is modulated in the low frequency range. Due to SBS nature, jitter variations in the optical frequency do not affect the correlation spectra measured with resolution figures up to 300 Hz, without the need for optical delay line as in conventional homodyne correlation techniques.


Vascular abnormalities and deregulation of VEGF in Lkb1-deficient mice.
The LKB1 tumor suppressor gene, mutated in Peutz-Jeghers syndrome, encodes a serine/threonine kinase of unknown function. Here we show that mice with a targeted disruption of Lkb1 die at midgestation, with the embryos showing neural tube defects, mesenchymal cell death, and vascular abnormalities. Extraembryonic development was also severely affected; the mutant placentas exhibited defective labyrinth layer development and the fetal vessels failed to invade the placenta. These phenotypes were associated with tissue-specific deregulation of vascular endothelial growth factor (VEGF) expression, including a marked increase in the amount of VEGF messenger RNA. Moreover, VEGF production in cultured Lkb1(-/-) fibroblasts was elevated in both normoxic and hypoxic conditions. These findings place Lkb1 in the VEGF signaling pathway and suggest that the vascular defects accompanying Lkb1 loss are mediated at least in part by VEGF.


Prognostic Value of Hypochloremia in Critically Ill Patients With Decompensated Cirrhosis.
Cirrhosis is frequently complicated by electrolyte disturbances, with prior studies primarily focused on the importance of hyponatremia. Emerging evidence on patients with chronic heart failure and chronic kidney disease has identified hypochloremia as an independent predictor for mortality. This study aimed to investigate the prognostic value of serum chloride and its association with mortality in cirrhotic patients. Retrospective cohort study. The medical ICU at Parkland Memorial Hospital, a tertiary care public health system in Dallas, Texas. Adult patients with confirmed diagnosis of decompensated cirrhosis who were admitted to the ICU between March 2015 and March 2017. None. Kaplan-Meier analysis and multivariable Cox proportional hazard ratio models were performed to determine the impact of hypochloremia on 180-day mortality. Of the 389 enrolled patients, 133 (34.2%) died within 180 days of ICU admission. Patients with hypochloremia had higher 180-day mortality than those with normochloremia (45.2% vs 26.7%; p < 0.0001). Cumulative survival via the Kaplan-Meier method was significantly lower in the hypochloremic group. Serum chloride was independently associated with 180-day mortality with multivariable adjustment (hazard ratio, 0.95; 95% CI, 0.93-0.98; p = 0.001) or after adjusting for Model for End-stage Liver Disease or Sequential Organ Failure Assessment. Contrarily, the inverse association between serum sodium and mortality no longer existed in all multivariable models. Serum chloride is independently and inversely associated with short-term mortality in critically ill cirrhotic patients. Hypochloremia, but not hyponatremia, remained associated with mortality with multivariable analyses, suggesting that hypochloremia may account for the mortality risk previously attributed to hyponatremia. These findings signify the prognostic value of serum chloride and potential inclusion of chloride into future cirrhosis prognostic scores.


Decoupling between water column oxygenation and benthic phosphate dynamics in a shallow eutrophic estuary.
Estuaries are crucial biogeochemical filters at the land-ocean interface that are strongly impacted by anthropogenic nutrient inputs. Here, we investigate benthic nitrogen (N) and phosphorus (P) dynamics in relation to physicochemical surface sediment properties and bottom water mixing in the shallow, eutrophic Peel-Harvey Estuary. Our results show the strong dependence of sedimentary P release on Fe and S redox cycling. The estuary contains surface sediments that are strongly reducing and act as net P source, despite physical sediment mixing under an oxygenated water column. This decoupling between water column oxygenation and benthic P dynamics is of great importance to understand the evolution of nutrient dynamics in marine systems in response to increasing nutrient loadings. In addition, the findings show that the relationship between P burial efficiency and bottom water oxygenation depends on local conditions; sediment properties rather than oxygen availability may control benthic P recycling. Overall, our results illustrate the complex response of an estuary to environmental change because of interacting physical and biogeochemical processes.


A genome-wide association study of IgM antibody against phosphorylcholine: shared genetics and phenotypic relationship to chronic lymphocytic leukemia.
Phosphorylcholine (PC) is an epitope on oxidized low-density lipoprotein (oxLDL), apoptotic cells and several pathogens like Streptococcus pneumoniae. Immunoglobulin M against PC (IgM anti-PC) has the ability to inhibit uptake of oxLDL by macrophages and increase clearance of apoptotic cells. From our genome-wide association studies (GWASs) in four European-ancestry cohorts, six single nucleotide polymorphisms (SNPs) in 11q24.1 were discovered (in 3002 individuals) and replicated (in 646 individuals) to be associated with serum level of IgM anti-PC (the leading SNP rs35923643-G, combined β = 0.19, 95% confidence interval 0.13-0.24, P = 4.3 × 10-11). The haplotype tagged by rs35923643-G (or its proxy SNP rs735665-A) is also known as the top risk allele for chronic lymphocytic leukemia (CLL), and a main increasing allele for general IgM. By using summary GWAS results of IgM anti-PC and CLL in the polygenic risk score (PRS) analysis, PRS on the basis of IgM anti-PC risk alleles positively associated with CLL risk (explained 0.6% of CLL variance, P = 1.2 × 10-15). Functional prediction suggested that rs35923643-G might impede the binding of Runt-related transcription factor 3, a tumor suppressor playing a central role in the immune regulation of cancers. Contrary to the expectations from the shared genetics between IgM anti-PC and CLL, an inverse relationship at the phenotypic level was found in a nested case-control study (30 CLL cases with 90 age- and sex-matched controls), potentially reflecting reverse causation. The suggested function of the top variant as well as the phenotypic association between IgM anti-PC and CLL risk needs replication and motivates further studies.


Lipoprotein[a] as a risk factor for preclinical atherosclerosis.
Elevated mean levels of lipoprotein[a] (Lp[a]) have been associated with symptomatic cardiovascular diseases such as clinically manifest myocardial infarction (MI), coronary artery disease, restenosis of coronary artery vein grafts after bypass, and a family history of MI. Associations of Lp[a] with arterial wall thickening in asymptomatic individuals previously have not been addressed and are evaluated in this report among participants of the Atherosclerosis Risk in Communities (ARIC) Study. Intima-media wall thickening in the extracranial carotid arteries was assessed noninvasively with B-mode ultrasonography; Lp[a] was measured as its total protein component. Individuals with wall thickening > or = 90th percentile of the population maximum far-wall thickness were pair matched to participants < 75th percentile of wall thickness by race, gender, center, 10-year age group, and time of examination. These selection criteria yielded 492 matched pairs, with 395 white pairs and 97 black pairs. The mean Lp[a] protein level for all black participants was 174.6 micrograms/mL compared with 77.8 micrograms/mL for whites. Conditional logistic regression analysis for the association of Lp[a] with case-control status yielded a statistically significant prevalence odds ratio (OR) estimate of 1.49, based on a 1-SD difference in Lp[a] protein, after adjusting for age, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, fibrinogen, hypertension, and cigarette smoking. None of these risk factors significantly altered the OR, in agreement with reports that Lp[a] is unaffected by environmental influences. In addition, no differential effect of Lp[a] protein on case-control status (effect modification) was observed by race, gender, low-density lipoprotein cholesterol, or fibrinogen in this population.(ABSTRACT TRUNCATED AT 250 WORDS)


Crystal structure of the Escherichia coli 23S rRNA:m5C methyltransferase RlmI (YccW) reveals evolutionary links between RNA modification enzymes.
Methylation is the most common RNA modification in the three domains of life. Transfer of the methyl group from S-adenosyl-l-methionine (AdoMet) to specific atoms of RNA nucleotides is catalyzed by methyltransferase (MTase) enzymes. The rRNA MTase RlmI (rRNA large subunit methyltransferase gene I; previously known as YccW) specifically modifies Escherichia coli 23S rRNA at nucleotide C1962 to form 5-methylcytosine. Here, we report the crystal structure of RlmI refined at 2 A to a final R-factor of 0.194 (R(free)=0.242). The RlmI molecule comprises three domains: the N-terminal PUA domain; the central domain, which resembles a domain previously found in RNA:5-methyluridine MTases; and the C-terminal catalytic domain, which contains the AdoMet-binding site. The central and C-terminal domains are linked by a beta-hairpin structure that has previously been observed in several MTases acting on nucleic acids or proteins. Based on bioinformatics analyses, we propose a model for the RlmI-AdoMet-RNA complex. Comparative structural analyses of RlmI and its homologs provide insight into the potential function of several structures that have been solved by structural genomics groups and furthermore indicate that the evolutionary paths of RNA and DNA 5-methyluridine and 5-methylcytosine MTases have been closely intertwined.


Effect of alterations of key active site residues in O6-alkylguanine-DNA Alkyltransferase on its ability to modulate the genotoxicity of 1,2-dibromoethane.
The production of mutations and the reduction in survival of cells treated with alpha,omega-dihaloalkanes is greatly enhanced by the presence of O6-alkylguanine-DNA alkyltransferase (AGT), a DNA repair protein that removes O6-alkylguanine adducts from DNA [Liu, L., Hachey, D. L., Valadez, G., Williams, K. M., Guengerich, F. P., Loktionova, N. A., Kanugula, S., and Pegg, A. E. (2004) J. Biol. Chem. 279, 4250-4259]. The effects of alterations to key residues in the active site of AGT were studied using AGTs with point mutations. It was found that mutants of AGT at positions Tyr114, Arg128, Pro140, Gly156, Gly160, and Tyr158 did not bring about the increase in genotoxicity of 1,2-dibromoethane seen with wild-type AGT, although these mutants, with the exception of those at Tyr114 and Arg128, are known to have sufficient AGT repair function to be able to protect cells from alkylating agents. The R128A mutant was able to react with 1,2-dibromoethane at the Cys145 acceptor site, but the resulting AGT-Cys145S-(CH2)2Br was much less able to produce a covalent adduct with DNA. This result is explained by the need for AGT to induce a structural change in the DNA "flipping" of a guanine nucleotide into the substrate binding pocket where Cys145 is located since the side chain of residue Arg128 plays a critical role in this reaction. Point mutations in AGT at the other sites (Y114A, P140K, and Y158H) reduced the ability of the protein to react with 1,2-dibromoethane as measured by the loss of activity. These results were confirmed by MS analysis of the tryptic peptide that contains the modified Cys145. There was no change in the stability of the AGT-Cys145S-(CH2)2Br intermediate formed in mutants Y158H and P140K. The reaction was studied in detail with mutant P140K using dihaloalkanes of different length; no effect of the mutations was seen with dibromomethane, but an enhanced difference was observed with 1,3-dibromopropane and 1,5-dibromopentane. These results show that even slight alterations in the active site pocket of AGT that do not prevent its ability to protect cells from alkylating agents can block the paradoxical enhancement of the genotoxicity of the larger alpha,omega-dihaloalkanes by reducing the reaction with Cys145.


Serotonin stimulates protein tyrosyl phosphorylation and vascular contraction via tyrosine kinase.
Serotonin (5-HT, 5-hydroxytryptamine) is a mitogen in vascular smooth muscle and vascular reactivity to 5-HT is significantly enhanced in hypertension and atherosclerosis. We have tested the hypothesis that tyrosine kinases, enzymes important for mitogenesis, may play a role in 5-HT-induced vascular smooth muscle contractility. Helical strips of rat carotid artery and aorta denuded of endothelium were mounted in tissue baths for measurement of contractile force. The tyrosine kinase inhibitor genistein (5 x 10(-6) M) decreased the potency of 5-HT approximately 4-fold and reduced maximal contraction to 5-HT in carotid arterial strips denuded of endothelium (58% control). Genistein's inactive congener daidzein (5 x 10(-6) M) did not reduce maximal contraction to 5-HT in carotid arteries but did shift the 5-HT concentration response curve 3-fold to the right. Tyrphostin 23 (5 x 10(-5) M), another tyrosine kinase inhibitor, decreased the potency of 5-HT 4-fold and reduced the maximal contraction to 5-HT in the carotid artery (10% control). Contractions induced by phorbol-12,13-dibutyrate (10(-9) to 10(-5) M) were not reduced or shifted by either tyrosine kinase inhibitor, indicating that phorbolester-sensitive protein kinase C isoforms were not affected. KCl-induced contraction was shifted 2-fold and the maximum significantly inhibited by tyrphostin 23 (38.6% control) but not genistein or daidzein, indicating that tyrphostin 23 but not genistein may inhibit voltage-gated calcium channels to reduce contractility. Western blot analysis using antiphosphotyrosine antibody confirmed that 5-HT produced a time- and concentration-dependent increase in the phosphotyrosine immunoreactivity of a 42-kD protein in cultured aortic smooth muscle cells. Lysate immunoprecipitation with an antimitogen-activated-protein (MAP)-kinase antibody indicated that the 42-kD protein was most likely a MAP kinase. 5-HT (10(-5) M) stimulated contraction and increased antiphosphotyrosine immunoreactivity in whole aorta mounted in tissue baths. Importantly, aortic contraction to 5-HT was shifted (5-fold rightward) and reduced (69% control) by genistein but not daidzein. These findings demonstrate that (1) tyrosine kinase activation may partially mediate contractility to 5-HT in arterial smooth muscle, (2) tyrphostin 23 is somewhat nonselective and (3) 5-HT stimulates tyrosine kinase as documented by increased tyrosyl phosphorylation of proteins in cultured aortic smooth muscle cells and aortic tissue in active contraction of 5-HT. These findings have significant implications not only in understanding a novel pathway of 5-HT signal transduction but also in vascular diseases in which growth and/or contractility to 5-HT is increased (e.g. hypertension, atherosclerosis).


Understanding visualization: a formal approach using category theory and semiotics.
This paper combines the vocabulary of semiotics and category theory to provide a formal analysis of visualization. It shows how familiar processes of visualization fit the semiotic frameworks of both Saussure and Peirce, and extends these structures using the tools of category theory to provide a general framework for understanding visualization in practice, including: Relationships between systems, data collected from those systems, renderings of those data in the form of representations, the reading of those representations to create visualizations, and the use of those visualizations to create knowledge and understanding of the system under inspection. The resulting framework is validated by demonstrating how familiar information visualization concepts (such as literalness, sensitivity, redundancy, ambiguity, generalizability, and chart junk) arise naturally from it and can be defined formally and precisely. This paper generalizes previous work on the formal characterization of visualization by, inter alia, Ziemkiewicz and Kosara and allows us to formally distinguish properties of the visualization process that previous work does not.


Process Optimization of Injection Overmolding Structural Electronics with Regard to Film Distortion.
The integration of structural electronics in injection-molded parts is a challenging step. The films-comprising of laminated stacks with electronics-are exposed to shear stresses and elevated temperatures by the molten thermoplastic. Hence, molding settings have a significant impact on the successful, damage-free manufacturing of such parts. In this paper, test films with polycarbonate (PC) sheets as outer and two different thermoplastic polyurethanes (TPUs) as middle layers incorporating conductive tracks on a flexible printed circuit board (flexPCB) are manufactured and overmolded with PC. Parameter studies investigating the influence of the melt temperature, mold temperature, injection speed and used TPU layer were performed. The molded parts were inspected visually and compared with a numerical simulation using injection molding software. A shear distortion factor for the TPU layer was derived based on the simulations that linked the shear stresses with the injection time and the softening (melting) of the TPU. The distortion of the films was found to reduce with higher melt temperature, lower mold temperature and faster injection speed. Films using the TPU with the higher melting temperature yielded significantly better results. Moreover, distortion on the films reduced with the increasing distance to the gate and a larger cavity thickness was found to be beneficial. All those relations could be correlated with the shear distortion factor.


Assessing the potential for post-copulatory sexual selection in elasmobranchs.
This review highlights the potential role that post-copulatory sexual selection plays in elasmobranch reproductive systems and the utility of this group to further understanding of evolutionary responses to the post-copulatory processes of sperm competition and cryptic female choice. The growing genetic evidence for female multiple mating (polyandry) in elasmobranchs is summarized. While polyandry appears to be common in this group, rates of multiple paternity are highly variable between species suggesting that there is large variance in the strength of post-copulatory sexual selection among elasmobranchs. Possible adaptations of traits important for post-copulatory sexual selection are then considered. Particular emphasis is devoted to explore the potential for sperm competition and cryptic female choice to influence the evolution of testes size, sperm morphology, genital morphology and sperm storage organs. Finally, it is argued that future work should take advantage of the wealth of information on these reproductive traits already available in elasmobranchs to gain a better understanding of how post-copulatory sexual selection operates in this group.


Communication strategies and cultural issues in the delivery of bad news.
Good communication is a fundamental skill for all palliative care clinicians. Patients present with varied desires, beliefs, and cultural practices, and navigating these issues presents clinicians with unique challenges. This article provides an overview of the evidence for communication strategies in delivering bad news and discussing advance care planning. In addition, it reviews the literature regarding cultural aspects of care for terminally ill patients and their families and offers strategies for engaging them. Through good communication practices, clinicians can help to avoid conflict and understand patients' desires for end of life care.


A novel histological grading scheme for placental malaria applied in areas of high and low malaria transmission.
Plasmodium falciparum-infected erythrocytes sequester in the placenta and elicit an inflammatory response that is harmful to both fetus and mother. Histologic measurements during placental malaria might provide surrogate end points for interventional trials, but existing histologic schemes capture limited complexity and are not consistently used among study sites. Using frozen-section histologic evaluation in Tanzania (high-transmission area), we established a novel grading scheme to separately quantify inflammation and pigment deposition during placental malaria (n = 102). To generalize this method, formalin-fixed, paraffin-embedded placental samples from Karen women in Thailand (low-transmission area) were selected from among women with documented antenatal parasitemia who were near term (n = 18). In the Tanzanian cohort, the inflammation and pigment-deposition scores were independently associated with birth weight, and the inflammation score was associated with chemokine levels. In the smaller cohort from Thailand, both inflammation and pigment scores were associated with birth weight, and the pigment score had an inverse trend with the number of antenatal clinic visits. This semiquantitative pathological grading scheme is simple to implement and captures information that is associated with outcomes in Asia and Africa; therefore, it should facilitate the comparison and standardization of results among clinical trials across areas of differing endemicity.


Use of pain-behavioral assessment tools in the nursing home: expert consensus recommendations for practice.
Many tools are available for the assessment of pain in nonverbal older adults; however, guidelines are needed to help clinicians select the proper instrument for use in the nursing home setting. This article describes a project to identify clinically useful pain-behavioral assessment tools that have undergone sufficient psychometric testing. Phase 1 of the project included a comprehensive review and critique of currently available tools. In Phase 2 the National Nursing Home Pain Collaborative developed criteria to evaluate an updated list of tools and then rated 14 tools using these criteria. As a result, two tools were recommended as most representative of current state of the science, most clinically relevant, and practically applicable to integrate into everyday practice and support adherence to regulatory guidelines. Such recommendations for selection of best-available pain assessment tools are a cornerstone for clinicians in regard to managing pain of nursing home residents who, due to dementia, are unable to self-report pain.


Evaluation for association between indwelling urethral catheter placement and risk of recurrent urethral obstruction in cats.
OBJECTIVE To test for an association between indwelling urethral catheter placement in cats with urethral obstruction (UO) and the short-term (30-day) risk of recurrent urethral obstruction (RUO). DESIGN Prospective cohort study. ANIMALS 107 client-owned male cats with UO. PROCEDURES Owners were offered standard care for their cats, including hospitalization, placement of an indwelling urethral catheter, IV fluid therapy, and other supportive treatments (inpatient group). One-time catheterization and outpatient care were offered (outpatient group) if standard care was declined. Data regarding signalment, measures of metabolic compromise and urinalysis findings at enrollment, catheterization-related variables, and supportive treatments of interest were collected. Risk of RUO ≤ 30 days after urethral catheter removal was determined for the outpatient vs inpatient group by OR and 95% confidence interval calculation. Other variables were compared between cats that did and did not develop RUO with Fisher exact and trend tests. RESULTS 91 cats completed the study; 19 (5/46 [11%] inpatients and 14/45 [31%] outpatients) developed RUO. Risk of RUO was significantly greater for cats of the outpatient group (OR, 3.7; 95% confidence interval, 1.2 to 11.4). Among inpatients, increasingly abnormal urine color at the time of catheter removal was significantly associated with RUO. No other significant associations were identified. CONCLUSIONS AND CLINICAL RELEVANCE Hospitalization and indwelling catheterization significantly reduced the risk for RUO ≤ 30 days after treatment for the population studied. Results suggested that removal of an indwelling catheter before urine appears grossly normal may be associated with development of RUO. One-time catheterization with outpatient care was inferior to the standard care protocol but was successful in many cats and may be a reasonable alternative when clients cannot pursue standard care.


Redox-regulation and life-history trade-offs: scavenging mitochondrial ROS improves growth in a wild bird.
It has been proposed that animals usually restrain their growth because fast growth leads to an increased production of mitochondrial reactive oxygen species (mtROS), which can damage mitochondrial DNA and promote mitochondrial dysfunction. Here, we explicitly test whether this occurs in a wild bird by supplementing chicks with a mitochondria-targeted ROS scavenger, mitoubiquinone (mitoQ), and examining growth rates and mtDNA damage. In the yellow-legged gull Larus michahellis, mitoQ supplementation increased the early growth rate of chicks but did not reduce mtDNA damage. The level of mtDNA damage was negatively correlated with chick mass, but this relationship was not affected by the mitoQ treatment. We also found that chick growth was positively correlated with both mtDNA copy number and the mitochondrial enzymatic activity of citrate synthase, suggesting a link between mitochondrial content and growth. Additionally, we found that MitoQ supplementation increased mitochondrial content (in males), altered the relationship between mtDNA copy number and damage, and downregulated some transcriptional pathways related to cell rejuvenation, suggesting that scavenging mtROS during development enhanced growth rates but at the expense of cellular turnover. Our study confirms the central role of mitochondria modulating life-history trade-offs during development by other mechanisms than mtROS-inflicted damage.


Proteomic analysis of human transplanted submandibular gland in patients with epiphora after transplantation.
Submandibular gland autotransplantation is effective for treating severe dry eye syndrome. However, more than 40% of patients show epiphora within 3-6 months after treatment. The mechanism underlying the hypersecretion in epiphora remains to be elucidated for developing novel interventions. Since salivary gland secretion is dependent on a variety of proteins, we analyzed the changes in protein expression in transplanted glands of epiphora patients with 2-D gel electrophoresis and electrospray ionization quadrupole/time-of-flight mass spectrometry and evaluated their possible roles in epiphora. There were 23 proteins that showed altered expression in the glands of epiphora patients, 15 being up-expressed and 8 being down-expressed. The expression of secretory proteins was decreased in these glands, including alpha-amylase, cystatin S, SA, and SN. In contrast, cytoskeletal proteins were all up-regulated, including actin and vimentin. Immunofluorescence revealed that the intensity ratio of F-actin in apical and lateral cytoplasm to total F-actin in acini was decreased in the glands of epiphora patients. Carbachol stimulation induced a similar redistribution of F-actin in the control glands. Phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) was increased in both carbachol-stimulated and epiphora glands. Preincubation of submandibular glands with ERK1/2 inhibitors PD98059 or U0126 inhibited carbachol-induced F-actin redistribution. These results indicated that differentially expressed proteins participated in the hypersecretion of transplanted submandibular glands and the redistribution of F-actin might be involved in this hypersecretion in an ERK1/2-dependent manner.


Nanoparticle-Aided Characterization of Arterial Endothelial Architecture during Atherosclerosis Progression and Metabolic Therapy.
Atherosclerosis is associated with a compromised endothelial barrier, facilitating the accumulation of immune cells and macromolecules in atherosclerotic lesions. In this study, we investigate endothelial barrier integrity and the enhanced permeability and retention (EPR) effect during atherosclerosis progression and therapy in Apoe-/- mice using hyaluronan nanoparticles (HA-NPs). Utilizing ultrastructural and en face plaque imaging, we uncover a significantly decreased junction continuity in the atherosclerotic plaque-covering endothelium compared to the normal vessel wall, indicative of disrupted endothelial barrier. Intriguingly, the plaque advancement had a positive effect on junction stabilization, which correlated with a 3-fold lower accumulation of in vivo administrated HA-NPs in advanced plaques compared to early counterparts. Furthermore, by using super-resolution and correlative light and electron microscopy, we trace nanoparticles in the plaque microenvironment. We find nanoparticle-enriched endothelial junctions, containing 75% of detected HA-NPs, and a high HA-NP accumulation in the endothelium-underlying extracellular matrix, which suggest an endothelial junctional traffic of HA-NPs to the plague. Finally, we probe the EPR effect by HA-NPs in the context of metabolic therapy with a glycolysis inhibitor, 3PO, proposed as a vascular normalizing strategy. The observed trend of attenuated HA-NP uptake in aortas of 3PO-treated mice coincides with the endothelial silencing activity of 3PO, demonstrated in vitro. Interestingly, the therapy also reduced the plaque inflammatory burden, while activating macrophage metabolism. Our findings shed light on natural limitations of nanoparticle accumulation in atherosclerotic plaques and provide mechanistic insight into nanoparticle trafficking across the atherosclerotic endothelium. Furthermore, our data contribute to the rising field of endothelial barrier modulation in atherosclerosis.


Examining the relationship between physical and sexual violence and psychosocial health in young people living with HIV in rural South Africa.
Experiences of violence during youth contravene young people's rights and increase the risk of depression and poor human immunodeficiency virus (HIV) care outcomes among youth living with HIV (YLWH). Intervention targets for mitigating the negative psychosocial effects of violence are needed, particularly in areas like rural South Africa where violence remains pervasive and mental healthcare is limited. This study aims to quantify the associations between physical and sexual violence and depressive symptoms in YLWH in rural South Africa and explore the modification of these associations by key measures of psychosocial well-being. We conducted a cross-sectional survey among 362 YLWH ages 12 to 24 in rural South Africa to ascertain participants' history of physical and sexual violence, current depressive symptoms (Center for Epidemiological Studies-Depression Scale) and levels of social support (Medical Outcomes Social Support Scale), resilience (Conner-Davidson Resilience Scale) and self-esteem (Rosenberg Self-Esteem Scale). Log-binomial regression was used to estimate the association between history of physical or sexual violence and clinically meaningful depressive symptoms (scores ≥ 16). Effect measure modification by high versus low resilience, social support and self-esteem was assessed using likelihood ratio tests (α = 0.20). A total of 334 individuals with a median age of 21 (interquartile range: 16 to 23) were included in this analysis. Most participants were female (71.3%), single (81.4%) and attending school (53.0%). Ninety-four participants (28.1%) reported a history of physical or sexual violence and 92 individuals (27.5%) had clinically meaningful depressive symptoms. Meaningful depressive symptoms were significantly higher among participants with a history of physical or sexual violence as compared to those with no history of violence (adjusted prevalence ratio: 2.01; 95% CI: 1.43, 2.83). However, this association was significantly modified by social support (p = 0.04) and self-esteem (p = 0.02). In this setting, the prevalence of meaningful depressive symptoms was significantly higher among YLWH with a history of physical or sexual violence as compared to those without a history of violence. However, higher levels of self-esteem or social support appeared to mitigate this association. Programmes to improve self-esteem and social support for youth have the potential to minimize depressive symptoms in YLWH who have experienced physical or sexual violence.


The impact of bright artificial white and 'blue-enriched' light on sleep and circadian phase during the polar winter.
Delayed sleep phase (and sometimes free-run) is common in the Antarctic winter (no natural sunlight) and optimizing the artificial light conditions is desirable. This project evaluated sleep when using 17,000 K blue-enriched lamps compared with standard white lamps (5000 K) for personal and communal illumination. Base personnel, 10 males, five females, 32.5±8 years took part in the study. From 24 March to 21 September 2006 light exposure alternated between 4-5-week periods of standard white (5000 K) and blue-enriched lamps (17,000 K), with a 3-week control before and after extra light. Sleep and light exposure were assessed by actigraphy and sleep diaries. General health (RAND 36-item questionnaire) and circadian phase (urinary 6-sulphatoxymelatonin rhythm) were evaluated at the end of each light condition. Direct comparison (rmanova) of blue-enriched light with white light showed that sleep onset was earlier by 19 min (P=0.022), and sleep latency tended to be shorter by 4 min (P=0.065) with blue-enriched light. Analysing all light conditions, control, blue and white, again provided evidence for greater efficiency of blue-enriched light compared with white (P<0.05), but with the best sleep timing, duration, efficiency and quality in control natural light conditions. Circadian phase was earlier on average in midwinter blue compared with midwinter white light by 45 min (P<0.05). Light condition had no influence on general health. We conclude that the use of blue-enriched light had some beneficial effects, notably earlier sleep, compared with standard white light during the polar winter.


Molecular Hydrogen Application in Stroke: Bench to Bedside.
Stroke is a major cause of mortality and morbidity worldwide. Effective treatments are limited. Molecular hydrogen is emerging as a novel medical gas with therapeutic potential for various neurological diseases, including stroke. We reviewed the experimental and clinical findings of the effects of molecular hydrogen therapy in stroke patients and models. The underlying neuroprotective mechanisms against stroke pathology were also discussed.


Risk assessment by monitoring expression levels of partial tandem duplications in the MLL gene in acute myeloid leukemia during therapy.
A partial tandem duplication within the MLL-gene (MLL-PTD) can be found in 8% of all patients with karyotypically normal acute myeloid leukemia (AML), a group in which polymerase chain reaction-(PCR) based minimal residual disease analysis has not, so far, been possible. A sensitive real-time PCR assay to quantify MLL-PTD transcripts was established and expression ratios assessed in diagnostic and follow-up samples. The prognostic significance of MLL-PTD expression levels was evaluated in 145 MLL-PTD positive patients at diagnosis and in 44 patients during and after treatment. Paired samples from 16 patients evaluated at diagnosis and relapse for the presence of the MLL-PTD were analyzed in parallel and all samples were positive at both time points. Overall, 173 samples from 44 patients were analyzed during follow-up (median sample number: 4/patient (range 2-17)). Nineteen patients were evaluable for MRD within the first 2 months, 15 patients within 4 months, and 19 patients within 6 months after the start of therapy. A >or= 2 log reduction of MLL-PTD expression in comparison to < 2 log reduction within 2, 4, and 6 months after start of therapy was found to be significantly associated with longer overall survival (p=0.029, p=0.007, and p=0.022, respectively). A molecular relapse was detected in 2 cases, in each case preceeding clinical manifestation by 35 days. These data suggest that MLL-PTD is a stable marker and can be used as a prognostically important marker of MRD in patients with karyotypically normal AML.


A critique of the standardization of the Miller Assessment for Preschoolers.
The Miller Assessment for Preschoolers (MAP) is a standardized screening instrument for preschoolers who are at risk for moderate to severe academic problems. A review of the procedures used to standardize the MAP found that it is a well-developed screening instrument that exhibits the qualities of good item discrimination, good test structure and content, good data collection on normal subjects, and good inter-rater and test-retest reliability. The MAP is effective in screening normal children who fall in the lowest 20th percentile. Since the intent of the MAP is to screen preschool children with moderate to severe pre-academic problems, further reliability and validity studies described in this paper need to be conducted before the MAP can be used effectively in screening children with delays. Caution should be used in categorizing normal children as "high risk" or suspect until decision validity studies have been completed.


Ventilation and breathlessness on maximal exercise in patients with interstitial lung disease after local anaesthetic aerosol inhalation.
1. The ventilatory response to maximal incremental exercise and the accompanying sensation of breathlessness were studied after the inhalation of 0.9% sodium chloride (saline) and 5% bupivacaine aerosols in six patients with interstitial lung disease. 2. The adequacy of airway anaesthesia induced by bupivacaine aerosol was confirmed by the absence of the cough reflex to 5% citric acid aerosol on completion of exercise. 3. All subjects first performed a trial exercise test to familiarize them with the procedure and to assess the degree of arterial oxygen desaturation on exercise. In subsequent tests, supplementary oxygen was given to maintain the saturation at 95% or above. 4. Airway anaesthesia had no effect on the ability to perform exercise as assessed by maximum workload, CO2 production or heart rate. No significant changes were seen on the pattern of breathing, minute ventilation or end-tidal PCO2 on exercise. There was, however, a small but statistically significant increase in ventilation related to CO2 production (VE/VCO2) at the end of exercise. 5. There was a tendency for breathlessness to be increased by airway anaesthesia but this did not reach statistical significance. 6. These results provide no evidence that vagal afferent activity is responsible for the abnormal ventilatory response to exercise in patients with interstitial lung disease. The perception of breathlessness in these patients was not diminished by anaesthesia of the airway.


Structure and dynamics of the tetrameric mnt repressor and a model for its DNA complex.
Abstract The tetrameric Mnt repressor of bacteriophage P22 consists of two dimeric DNA-binding domains and a tetramerization domain. The NOE and chemical shift data demonstrate that the structures of the domains in the wild-type repressor protein are similar to those of the separate domains, the three-dimensional structures of which have been determined previously. (15)N relaxation measurements show that the linker that connects the anti-parallel four-helix bundle with the two β-sheet DNA-binding dimers is highly flexible. No evidence was found for interactions between the distinct modules. The (15)N relaxation properties of the two domains differ substantially, confirming their structural independence. A model in which one two-stranded coiled coil of the four-helix bundle is attached to one N-terminal dimer is most consistent with the biochemical data and (15)N relaxation data. For the Mnt-DNA complex this geometry fits with a model in which the two β-sheet DNA-binding domains are bound at two successive major grooves of the Mnt operator and the tetramerization domain is packed between these two DNA-bound dimers. In such a model the two-fold symmetry axis of the four-helix bundle coincides with that of the operator sequence and the two bound dimers. Bending of the Mnt operator of approximately 30° upon binding of the tetramer, as measured by gel-shift assays, is in agreement with this model of the Mnt-DNA complex.


Experimental study on copper uptake capacity in the Mediterranean mussel (Mytilus galloprovincialis).
To investigate the effect of different sizes, sex, and exposure time on Cu uptake capacity, mussels Mytilus galloprovincialis of different shell sizes were exposed to different Cu concentrations in different aquariums. In another experiment, mussels were exposed to stable dissolved Cu for 6 days in the laboratory. All mussels tissue concentrations were analyzed using energy dispersive X-ray fluorescence (EDXRF) spectrometry. At the end of uptake, the rate of increase of Cu level in the soft tissues of mussels in different aquariums was 3.84-7.92 times higher than before exposure. While the results of Cu concentrations were negatively correlated with the shell sizes in the control and second groups (r control = -0.862, r second = -0.851 p < 0.05), this relation was not observed in the other groups (p > 0.05). Also, results showed no significant difference between male and female (p > 0.05). On the other hand, Cu concentration values in soft tissue were monitored daily and observed to be increasing up to the third day but afterwards to be descending, thus indicating a significant effect of the exposure time-related Cu uptake by mussels. Therefore, the exposure time to Cu metal of the mussel should be taken into account in the marine pollution investigations. In addition, by using the obtained Cu heavy metal concentration results, the heavy metal intake by the human population was calculated by taking into account daily mussel consumption. The results were examined for potential human health risks and discussed. These results would be helpful to understand factors controlling Cu accumulation in mussels.


Whole genome sequence analysis reveals the broad distribution of the RtxA type 1 secretion system and four novel putative type 1 secretion systems throughout the Legionella genus.
Type 1 secretion systems (T1SSs) are broadly distributed among bacteria and translocate effectors with diverse function across the bacterial cell membrane. Legionella pneumophila, the species most commonly associated with Legionellosis, encodes a T1SS at the lssXYZABD locus which is responsible for the secretion of the virulence factor RtxA. Many investigations have failed to detect lssD, the gene encoding the membrane fusion protein of the RtxA T1SS, in non-pneumophila Legionella, which has led to the assumption that this system is a virulence factor exclusively possessed by L. pneumophila. Here we discovered RtxA and its associated T1SS in a novel Legionella taurinensis strain, leading us to question whether this system may be more widespread than previously thought. Through a bioinformatic analysis of publicly available data, we classified and determined the distribution of four T1SSs including the RtxA T1SS and four novel T1SSs among diverse Legionella spp. The ABC transporter of the novel Legionella T1SS Legionella repeat protein secretion system shares structural similarity to those of diverse T1SS families, including the alkaline protease T1SS in Pseudomonas aeruginosa. The Legionella bacteriocin (1-3) secretion systems T1SSs are novel putative bacteriocin transporting T1SSs as their ABC transporters include C-39 peptidase domains in their N-terminal regions, with LB2SS and LB3SS likely constituting a nitrile hydratase leader peptide transport T1SSs. The LB1SS is more closely related to the colicin V T1SS in Escherichia coli. Of 45 Legionella spp. whole genomes examined, 19 (42%) were determined to possess lssB and lssD homologs. Of these 19, only 7 (37%) are known pathogens. There was no difference in the proportions of disease associated and non-disease associated species that possessed the RtxA T1SS (p = 0.4), contrary to the current consensus regarding the RtxA T1SS. These results draw into question the nature of RtxA and its T1SS as a singular virulence factor. Future studies should investigate mechanistic explanations for the association of RtxA with virulence.


Fumagillin prodrug nanotherapy suppresses macrophage inflammatory response via endothelial nitric oxide.
Antiangiogenesis has been extensively explored for the treatment of a variety of cancers and certain inflammatory processes. Fumagillin, a mycotoxin produced by Aspergillus fumigatus that binds methionine aminopeptidase 2 (MetAP-2), is a potent antiangiogenic agent. Native fumagillin, however, is poorly soluble and extremely unstable. We have developed a lipase-labile fumagillin prodrug (Fum-PD) that eliminated the photoinstability of the compound. Using αvβ3-integrin-targeted perfluorocarbon nanocarriers to deliver Fum-PD specifically to angiogenic vessels, we effectively suppressed clinical disease in an experimental model of rheumatoid arthritis (RA). The exact mechanism by which Fum-PD-loaded targeted nanoparticles suppressed inflammation in experimental RA, however, remained unexplained. We herein present evidence that Fum-PD nanotherapy indirectly suppresses inflammation in experimental RA through the local production of endothelial nitric oxide (NO). Fum-PD-induced NO activates AMP-activated protein kinase (AMPK), which subsequently modulates macrophage inflammatory response. In vivo, NO-induced AMPK activation inhibits mammalian target of rapamycin (mTOR) activity and enhances autophagic flux, as evidenced by p62 depletion and increased autolysosome formation. Autophagy in turn mediates the degradation of IkappaB kinase (IKK), suppressing the NF-κB p65 signaling pathway and inflammatory cytokine release. Inhibition of NO production by N(G)-nitro-L-arginine methyl ester (L-NAME), a nitric oxide synthase inhibitor, reverses the suppression of NF-κB-mediated inflammatory response induced by Fum-PD nanotherapy. These unexpected results uncover an activity of Fum-PD nanotherapy that may be further explored in the treatment of angiogenesis-dependent diseases.


iRest Meditation for Older Adults with Depression Symptoms: A Pilot Study.
Older adults, a rapidly growing population in the United States, have fewer physiological reserves and are more likely to be affected by stress, making them especially susceptible to depression symptoms. Meditation offers promising potential as an effective treatment; however, few studies have evaluated meditation interventions for this demographic. The objectives of this pilot study were to evaluate the feasibility and acceptability of an iRest meditation program in older adults with depression symptoms and to collect preliminary data on its effect on depression and depression-related symptoms compared to a vacation control. The study occurred at the Institute of Noetic Sciences EarthRise Retreat Center and participants' homes. Thirty generally healthy older adults, aged 55-90, with depression symptoms were recruited. Participants were randomly assigned to a 2-day retreat of either iRest meditation training or vacation. After the retreat, participants were asked to complete 20 minutes of home practice per day for 6 weeks; this consisted of either guided meditations (iRest) or music (vacation). Data were collected pre- and post-retreat and then 6 weeks later. Measures included depression-related variables (expectancy, depression symptoms, perceived stress, resilience, pain, sleep quality, and spirituality) and biomarkers (voice stress analysis, heart rate, heart rate variability). We found the iRest intervention for older adults with depression symptoms to be feasible and acceptable. Preliminary results at 6 weeks demonstrated improvements in sleep impairment in older adults compared to the control group and promising trends in improvements in depression symptoms and pain severity.


Analog-digital simulations of full conductance-based networks of spiking neurons with spike timing dependent plasticity.
We introduce and test a system for simulating networks of conductance-based neuron models using analog circuits. At the single-cell level, we use custom-designed analog circuits (ASICs) that simulate two types of spiking neurons based on Hodgkin-Huxley like dynamics: "regular spiking" excitatory neurons with spike-frequency adaptation, and "fast spiking" inhibitory neurons. Synaptic interactions are mediated by conductance-based synaptic currents described by kinetic models. Connectivity and plasticity rules are implemented digitally through a real time interface between a computer and a PCI board containing the ASICs. We show a prototype system of a few neurons interconnected with synapses undergoing spike-timing dependent plasticity (STDP), and compare this system with numerical simulations. We use this system to evaluate the effect of parameter dispersion on the behavior of small circuits of neurons. It is shown that, although the exact spike timings are not precisely emulated by the ASIC neurons, the behavior of small networks with STDP matches that of numerical simulations. Thus, this mixed analog-digital architecture provides a valuable tool for real-time simulations of networks of neurons with STDP. They should be useful for any real-time application, such as hybrid systems interfacing network models with biological neurons.


The Relationship Between Aerobic and Anaerobic Performance in Recreational Runners.
Research has indicated that combined aerobic and anaerobic training (concurrent training) may improve aerobic performance greater than aerobic training alone. The purpose of this investigation was to establish any associations between aerobic and anaerobic performance. Eleven participants (n = 11, age = 34.1 ± 13 years, VO2max = 58.4 ± 7.8) volunteered for this study. Participants were asked for endurance training experience (4.7 ± 3.7 years) and resistance training experience (4.1 ± 4.6 years). To meet training status, participants were to have a VO2max in the 80th percentile as per ACSM guidelines. The Bruce treadmill test was used to measure aerobic performance. In order to measure anaerobic performance, several tests were completed utilizing a force platform. A Pearson Product R Correlation Coefficient was calculated to determine correlations between variables. The results show significant correlation between VO2max and RFD (r = 0.68). Further analyses utilizing Cohen's effect size indicated a strong association between VO2max and peak force, as well as running efficiency and peak power, relative peak power, and power endurance. These results indicate an existing possibility that anaerobic performance measures such as RFD may have a positive relationship with aerobic performance measures such as VO2max. Therefore, it may be beneficial to integrate specific training components which focus on improving RFD as a method of improving running performance.


Lactococcal phage p2 ORF35-Sak3 is an ATPase involved in DNA recombination and AbiK mechanism.
Virulent phages of the Siphoviridae family are responsible for milk fermentation failures worldwide. Here, we report the characterization of the product of the early expressed gene orf35 from Lactococcus lactis phage p2 (936 group). ORF35(p2), also named Sak3, is involved in the sensitivity of phage p2 to the antiviral abortive infection mechanism AbiK. The localization of its gene upstream of a gene coding for a single-strand binding protein as well as its membership to a superfamily of single-strand annealing proteins (SSAPs) suggested a possible role in homologous recombination. Electron microscopy showed that purified ORF35(p2) form a hexameric ring-like structure that is often found in proteins with a conserved RecA nucleotide-binding core. Gel shift assays and surface plasmon resonance data demonstrated that ORF35(p2) interacts preferentially with single-stranded DNA with nanomolar affinity. Atomic force microscopy showed also that it preferentially binds to sticky DNA substrates over blunt ends. In addition, in vitro assays demonstrated that ORF35(p2) is able to anneal complementary strands. Sak3 also stimulates Escherichia coli RecA-mediated homologous recombination. Remarkably, Sak3 was shown to possess an ATPase activity that is required for RecA stimulation. Collectively, our results demonstrate that ORF35(p2) is a novel SSAP stimulating homologous recombination.


Determinants of timely initiation of breastfeeding among mothers in Goba Woreda, South East Ethiopia: a cross sectional study.
Although breastfeeding is universal in Ethiopia, ranges of regional differences in timely initiation of breastfeeding have been documented. Initiation of breastfeeding is highly bound to cultural factors that may either enhance or inhibit the optimal practices. The government of Ethiopia developed National Infant and Young Child Feeding Guideline in 2004 and behavior change communications on breast feeding have been going on since then. However, there is a little information on the practice of timely initiation of breast feeding and factors that predict these practices after the implementation of the national guideline. The objective of this study is to determine the prevalence and determinant factors of timely initiation of breastfeeding among mothers in Bale Goba District, South East Ethiopia. A community based cross sectional study was carried out from February to March 2010 using both quantitative and qualitative methods of data collection. A total of 608 mother infant pairs were selected using simple random sampling method and key informants for the in-depth interview were selected conveniently. Descriptive statistics, bivariate analysis and multivariable logistic regression analyses were employed to identify factors associated with timely initiation of breast feeding. The prevalence of timely initiation of breastfeeding was 52.4%. Bivariate analysis showed that attendance of formal education, being urban resident, institutional delivery and postnatal counseling on breast feeding were significantly associated with timely initiation of breastfeeding (P < 0.05). After adjust sting for other factors on the multivariable logistic model, being in the urban area [AOR: 4.1 (95%C.I: 2.31-7.30)] and getting postnatal counseling [AOR: 2.7(1.86-3.94)] were independent predictors of timely initiation of breastfeeding. The practice of timely initiation of breast feeding is low as nearly half the mothers did not start breastfeeding with one hour after delivery. The results suggest that breast feeding behavior change communication especially during the post natal period is critical in promoting optimal practice in the initiation of breast feeding. Rural mothers need special attention as they are distant from various information sources.


'I don't have any other choice': spouses' experiences of placing a partner in a care home for older people in Sweden.
The main aim of this paper is to consider the experiences of Swedish spouses who have placed a partner in a care home for older people. Data were gathered from semi-structural interviews with 14 spouses (11 wives and 3 husbands) who had been involved in a care home placement within the previous 6 months. The results reported here are from the first component of a larger grounded theory study, the aim of which is to explore, describe and understand the experience of care home placement from a variety of perspectives and to identify the implications for policy and practice in Sweden. The focus here is on the experience of spouses, relating to the decision-making process, the move into care and subsequent contact with the care home. Four themes emerged from the data - making the decision, making the move, adjusting to the move and reorientation. The results show a lack of planning for the elder person's entry to a care home, and professional dominance of this stage of the process. The largely ambivalent emotional responses to the move that spouses experience and the difficulties in initiating and sustaining relationships with staff in the home are discussed in the light of previous research.


Functional disturbances in the striatum by region-specific ablation of NMDA receptors.
To study the role of NMDA receptors in dopamine signaling of the striatum, the brain area that receives glutamatergic inputs from various cortical areas and most dopaminergic inputs, we generated striatum-specific NMDA receptor-deficient mice. The mutant pups showed reduced food intake and retarded growth starting at the second postnatal week and died on approximately postnatal day 20 (P20). The time course of postnatal lethality is similar to that of compound mutant, double knockout of dopamine D1/D2 receptors, or genetically engineered dopamine-deficient mouse. In vivo electrophysiological recordings in the mutant pups showed that frequencies in the range of gamma oscillation were reduced in the striatal circuits. Moreover, the number of functional dopamine receptors in the striatum as measured by D1- and D2-binding experiments was greatly diminished in the mutants as compared with control animals. A consequence of diminished dopamine binding in the striatum manifested in an increase of locomotor activity. The administration of D1/D2 agonists paradoxically reduced the hyperactivity of the mutant mice as compared with an increase in locomotor activity in control mice. These results demonstrate that the NMDA receptor plays an essential role in the integration of dopamine signaling in the striatum and that is required in behavioral function.


Limited genetic and antigenic diversity within parasite isolates used in a live vaccine against Theileria parva.
An infection and treatment protocol is used to vaccinate cattle against Theileria parva infection. Due to incomplete cross-protection between different parasite isolates, a mixture of three isolates, termed the Muguga cocktail, is used for vaccination. While vaccination of cattle in some regions provides high levels of protection, some animals are not protected against challenge with buffalo-derived T. parva. Knowledge of the genetic composition of the Muguga cocktail vaccine is required to understand how vaccination is able to protect against field challenge and to identify the potential limitations of the vaccine. The aim of the current study was to determine the extent of genetic and antigenic diversity within the parasite isolates that constitute the Muguga cocktail. High throughput multi-locus sequencing of antigen-encoding loci was performed in parallel with typing using a panel of micro- and mini-satellite loci. The former focused on genes encoding CD8(+) T cell antigens, believed to be relevant to protective immunity. The results demonstrate that each of the three component stocks of the cocktail contains limited parasite genotypic diversity, with single alleles detected at many gene/satellite loci and, moreover, that two of the components show a very high level of similarity. Thus, the vaccine incorporates very little of the genetic and antigenic diversity observed in field populations of T. parva. The presence of alleles at low frequency (<10%) within vaccine component populations also points to the possibility of variability in the content of vaccine doses and the potential for loss of allelic diversity during tick passage. The results demonstrate that there is scope to modify the content of the vaccine in order to enhance its diversity and thus its potential for providing broad protection. The ability to accurately quantify genetic diversity in vaccine component stocks will facilitate improved quality control procedures designed to ensure the long-term efficacy of the vaccine.


A computer-simulation study on the effects of MRI voxel dimensions on carotid plaque lipid-core and fibrous cap segmentation and stress modeling.
The benefits of a decreased slice thickness and/or in-plane voxel size in carotid MRI for atherosclerotic plaque component quantification accuracy and biomechanical peak cap stress analysis have not yet been investigated in detail because of practical limitations. In order to provide a methodology that allows such an investigation in detail, numerical simulations of a T1-weighted, contrast-enhanced, 2D MRI sequence were employed. Both the slice thickness (2 mm, 1 mm, and 0.5 mm) and the in plane acquired voxel size (0.62x0.62 mm2 and 0.31x0.31 mm2) were varied. This virtual MRI approach was applied to 8 histology-based 3D patient carotid atherosclerotic plaque models. A decreased slice thickness did not result in major improvements in lumen, vessel wall, and lipid-rich necrotic core size measurements. At 0.62x0.62 mm2 in-plane, only a 0.5 mm slice thickness resulted in improved minimum fibrous cap thickness measurements (a 2-3 fold reduction in measurement error) and only marginally improved peak cap stress computations. Acquiring voxels of 0.31x0.31 mm2 in-plane, however, led to either similar or significantly larger improvements in plaque component quantification and computed peak cap stress. This study provides evidence that for currently-used 2D carotid MRI protocols, a decreased slice thickness might not be more beneficial for plaque measurement accuracy than a decreased in-plane voxel size. The MRI simulations performed indicate that not a reduced slice thickness (i.e. more isotropic imaging), but the acquisition of anisotropic voxels with a relatively smaller in-plane voxel size could improve carotid plaque quantification and computed peak cap stress accuracy.


Enhanced trafficking of tetrameric Kv4.3 channels by KChIP1 clamping.
The cytoplamsic auxiliary KChIPs modulate surface expression and gating properties of Kv4 channels. Recent co-crystal structure of Kv4.3 N-terminus and KChIP1 reveals a clamping action of the complex in which a single KChIP1 molecule laterally binds two neighboring Kv4.3 N-termini at different locations, thus forming two contact interfaces involved in the protein-protein interaction. In the second interface, it functions to stabilize the tetrameric assembly, but the role it plays in channel trafficking remains elusive. In this study, we examined the effects of KChIP1 on Kv4 protein trafficking in COS-7 cells expressing EGFP-tagged Kv4.3 channels using confocal microscopy. Mutations either in KChIP1 (KChIP1 L39E-Y57A-K61A) or Kv4.3 (Kv4.3 E70A-F73E) that disrupt the protein-protein interaction within the second interface can reduce surface expression of Kv4 channel proteins. Kv4.3 C110A, the Zn2+ binding site mutation in T1 domain, that disrupts the tetrameric assembly of the channels can be rescued by WT KChIP1, but not the KChIP1 triple mutant. These results were further confirmed by whole cell current recordings in oocytes. Our findings show that key residues of second interface involved in stabilizing tetrameric assembly can regulate the channel trafficking, indicating an intrinsic link between tetrameric assembly and channel trafficking. The results also suggest that formation of octameric Kv4 and KChIP complex by KChIPs clamping takes place before their trafficking to final destination on the cell surface.


Biomarkers and Genetics in Peripheral Artery Disease.
Peripheral artery disease (PAD) is highly prevalent and there is considerable diversity in the initial clinical manifestation and disease progression among individuals. Currently, there is no ideal biomarker to screen for PAD, to risk stratify patients with PAD, or to monitor therapeutic response to revascularization procedures. Advances in human genetics have markedly enhanced the ability to develop novel diagnostic and therapeutic approaches across a host of human diseases, but such developments in the field of PAD are lagging. In this article, we will discuss the epidemiology, traditional risk factors for, and clinical presentations of PAD. We will discuss the possible role of genetic factors and gene-environment interactions in the development and/or progression of PAD. We will further explore future avenues through which genetic advances can be used to better our understanding of the pathophysiology of PAD and potentially find newer therapeutic targets. We will discuss the potential role of biomarkers in identifying patients at risk for PAD and for risk stratifying patients with PAD, and novel approaches to identification of reliable biomarkers in PAD. The exponential growth of genetic tools and newer technologies provides opportunities to investigate and identify newer pathways in the development and progression of PAD, and thereby in the identification of newer biomarkers and therapies.


Contribution of Anal Sex to HIV Prevalence Among Heterosexuals: A Modeling Analysis.
Anal intercourse is reported by many heterosexuals, and evidence suggests that its practice may be increasing. We estimated the proportion of the HIV burden attributable to anal sex in 2015 among heterosexual women and men in the United States. The HIV Optimization and Prevention Economics model was developed using parameter inputs from the literature for the sexually active U.S. population aged 13-64. The model uses differential equations to represent the progression of the population between compartments defined by HIV disease status and continuum-of-care stages from 2007 to 2015. For heterosexual women of all ages (who do not inject drugs), almost 28% of infections were associated with anal sex, whereas for women aged 18-34, nearly 40% of HIV infections were associated with anal sex. For heterosexual men, 20% of HIV infections were associated with insertive anal sex with women. Sensitivity analyses showed that varying any of 63 inputs by ±20% resulted in no more than a 13% change in the projected number of heterosexual infections in 2015, including those attributed to anal sex. Despite uncertainties in model inputs, a substantial portion of the HIV burden among heterosexuals appears to be attributable to anal sex. Providing information about the relative risk of anal sex compared with vaginal sex may help reduce HIV incidence in heterosexuals.


Usefulness of 124I PET/CT imaging to predict absorbed doses in patients affected by metastatic thyroid cancer and treated with 131I.
The aim of this work was the evaluation of the usefulness of 124I PET/CT sequential scans to predict absorbed doses to metastatic thyroid cancer in patients undergoing 131I therapy. From July 2011 until April 2012 8 patients affected by metastatic thyroid cancer were enrolled. Each patient underwent 4 PET/CT scans at 4, 24, 48, 72 h after the administration of about 74 MBq of 124I. Blood samples and whole body exposure measurements were obtained to calculate blood and red marrow doses. Activity concentrations and lesion volumes obtained from PET/CT images were used to evaluate tumour doses with MIRD formalism and spheres model. The average administered 131I therapeutic activity was 6475 MBq (range: 3700-9250 MBq). 124I PET/CT images showed, with a very good resolution, all 131I avid lesions detected by post therapy whole body scans. The average dose rates for blood, red marrow and lesions were respectively: 6.58E-02 ± 1.64E-02 mGy/MBq, 5.73E-02 ± 1.57E-02 mGy/MBq, 2.22E+01 ± 1.62E+01 mGy/MBq. Three out of eight patients did not show any uptake of 124I in all PET/CT scans, despite high level of TSH and CT detectable lesions. Post-therapy 131I whole body scan confirmed the absence of focal iodine uptake. Negative 124I PET/CT images probably could be used as predictive of real absence of iodine avidity, avoiding all toxicity from useless 131I therapy. A higher number of patients is necessary to validate these preliminary results and a project is ongoing to compare MIRD results to voxel dosimetry based on Monte Carlo simulation.