Near-unity broadband absorption designs for semiconducting nanowire arrays via localized radial mode excitation.
We report design methods for achieving near-unity broadband light absorption in sparse nanowire arrays, illustrated by results for visible absorption in GaAs nanowires on Si substrates. Sparse (<5% fill fraction) nanowire arrays achieve near unity absorption at wire resonant wavelengths due to coupling into 'leaky' radial waveguide modes of individual wires and wire-wire scattering processes. From a detailed conceptual development of radial mode resonant absorption, we demonstrate two specific geometric design approaches to achieve near unity broadband light absorption in sparse nanowire arrays: (i) introducing multiple wire radii within a small unit cell array to increase the number of resonant wavelengths, yielding a 15% absorption enhancement relative to a uniform nanowire array and (ii) tapering of nanowires to introduce a continuum of diameters and thus resonant wavelengths excited within a single wire, yielding an 18% absorption enhancement over a uniform nanowire array.


Attention, in and Out: Scalp-Level and Intracranial EEG Correlates of Interoception and Exteroception.
Interoception, the monitoring of visceral signals, is often presumed to engage attentional mechanisms specifically devoted to inner bodily sensing. In fact, most standardized interoceptive tasks require directing attention to internal signals. However, most studies in the field have failed to compare attentional modulations between internally- and externally-driven processes, thus probing blind to the specificity of the former. Here we address this issue through a multidimensional approach combining behavioral measures, analyses of event-related potentials and functional connectivity via high-density electroencephalography, and intracranial recordings. In Study 1, 50 healthy volunteers performed a heartbeat detection task as we recorded modulations of the heartbeat-evoked potential (HEP) in three conditions: exteroception, basal interoception (also termed interoceptive accuracy), and post-feedback interoception (sometimes called interoceptive learning). In Study 2, to evaluate whether key interoceptive areas (posterior insula, inferior frontal gyrus, amygdala, and somatosensory cortex) were differentially modulated by externally- and internally-driven processes, we analyzed human intracranial recordings with depth electrodes in these regions. This unique technique provides a very fine grained spatio-temporal resolution compared to other techniques, such as EEG or fMRI. We found that both interoceptive conditions in Study 1 yielded greater HEP amplitudes than the exteroceptive one. In addition, connectivity analysis showed that post-feedback interoception, relative to basal interoception, involved enhanced long-distance connections linking frontal and posterior regions. Moreover, results from Study 2 showed a differentiation between oscillations during basal interoception (broadband: 35-110 Hz) and exteroception (1-35 Hz) in the insula, the amygdala, the somatosensory cortex, and the inferior frontal gyrus. In sum, this work provides convergent evidence for the specificity and dynamics of attentional mechanisms involved in interoception.


rROP2(186-533): a novel peptide antigen for detection of IgM antibodies against Toxoplasma gondii.
Toxoplasma gondii infections are prevalent in a wide range of mammalian hosts including humans. Infection in pregnant women may cause the transmission of parasite to the fetus that makes serious problems. IgM antibodies against Toxoplasma (Toxo-IgM) have been believed to be significant indicators for both recently acquired and congenital toxoplasmosis. So far, however, there has not been any recognized protein of T. gondii that specifically reacts to IgM antibodies. Here, an antigen exclusively for detection of IgM antibodies screened by two-dimensional electrophoresis and mass spectrometry has been reported. The study identified 13 Toxoplasma proteins probed by IgG antibodies and one (rhpotry protein 2 [ROP2]) by IgM antibodies with human sera of Toxo-IgM(-)-IgG(+) and -IgM(+)-IgG(-), respectively, which had been prescreened by Toxo-IgM and -IgG commercial kits from the suspected cases. Following cloning, expression, and purification of the fragment of ROP2(186-533), an enzyme-linked immunosorbent assay with rROP2(186-533) to measure IgM and IgG antibodies was developed. As a result, 100%(48/48) of sera with Toxo-IgM(+)-IgG(-)showed positive Toxo-IgM but none of them (0%) showed positive Toxo-IgG when rROP2(186-533) was used as antigen. Neither Toxo-IgG nor Toxo-IgM antibodies were found when tested with 59 sera of Toxo-IgM(-)-IgG(+). These results indicate that rROP2(186-533) could be used as an antigen that specifically capture Toxo-IgM antibodies and may have a high potential in the serological diagnosis of both acute acquired and congenital toxoplasmosis.


Cl- fluxes related to fluid secretion by the rat parotid: involvement of Cl(-)-HCO3- exchange.
Muscarinic-induced 36Cl- and 86Rb+ (K+ substitute) fluxes were studied in rat parotid acini. Stimulation resulted in a rapid [half time (t1/2) less than 30 s] decrease in both Cl- and Rb+ content (approximately 50 and 30%, respectively) followed by a slower partial recovery (t1/2 approximately 3-4 min) to approximately 80% of resting levels for both ions. Cl- loss was inhibited by the venom of Leiurus quinquestriatus, which contains the maxi-K+ channel blocker charybdotoxin. Cl- recovery was blunted in the presence of bumetanide, an inhibitor of Na(+)-K(+)-Cl- cotransport, or on HCO3- removal and was completely blocked in the presence of bumetanide and 4,4' diisothiocyanostilbene-2,2' disulfonic acid (DIDS), an inhibitor of Cl(-)-HCO3- exchange. In HCO3(-)-containing medium a rapid (t1/2 less than 1 min), DIDS-inhibitable cytoplasmic alkalinization (approximately 0.4 pH unit) was observed in acini switched to a Cl(-)-free solution. This alkalinization was not seen in HCO3(-)-free medium but persisted in the absence of Na+, consistent with the presence of a potent Na(+)-independent Cl(-)-HCO3- exchanger. Kinetic studies indicated that the half-maximal effect of this exchanger for extracellular Cl- was approximately 18 mM. These results are consistent with the hypothesis that secretagogue-induced KCl loss by salivary acinar cells occurs via electrically coupled K+ and Cl- channels. In addition, they provide strong evidence that Cl- entry into, and thus fluid secretion by, these cells is mediated by both Cl(-)-HCO3- exchange and Na(+)-K(+)-Cl- cotransport.


Getting personalized cancer genome analysis into the clinic: the challenges in bioinformatics.
Progress in genomics has raised expectations in many fields, and particularly in personalized cancer research. The new technologies available make it possible to combine information about potential disease markers, altered function and accessible drug targets, which, coupled with pathological and medical information, will help produce more appropriate clinical decisions. The accessibility of such experimental techniques makes it all the more necessary to improve and adapt computational strategies to the new challenges. This review focuses on the critical issues associated with the standard pipeline, which includes: DNA sequencing analysis; analysis of mutations in coding regions; the study of genome rearrangements; extrapolating information on mutations to the functional and signaling level; and predicting the effects of therapies using mouse tumor models. We describe the possibilities, limitations and future challenges of current bioinformatics strategies for each of these issues. Furthermore, we emphasize the need for the collaboration between the bioinformaticians who implement the software and use the data resources, the computational biologists who develop the analytical methods, and the clinicians, the systems' end users and those ultimately responsible for taking medical decisions. Finally, the different steps in cancer genome analysis are illustrated through examples of applications in cancer genome analysis.


Patterns, determinants and models of woody plant diversity in China.
What determines large-scale patterns of species richness remains one of the most controversial issues in ecology. Using the distribution maps of 11 405 woody species in China, we compared the effects of habitat heterogeneity, human activities and different aspects of climate, particularly environmental energy, water-energy dynamics and winter frost, and explored how biogeographic affinities (tropical versus temperate) influence richness-climate relationships. We found that the species richness of trees, shrubs, lianas and all woody plants strongly correlated with each other, and more strongly correlated with the species richness of tropical affinity than with that of temperate affinity. The mean temperature of the coldest quarter was the strongest predictor of species richness, and its explanatory power for species richness was significantly higher for tropical affinity than for temperate affinity. These results suggest that the patterns of woody species richness mainly result from the increasing intensity of frost filtering for tropical species from the equator/lowlands towards the poles/highlands, and hence support the freezing-tolerance hypothesis. A model based on these results was developed, which explained 76-85% of species richness variation in China, and reasonably predicted the species richness of woody plants in North America and the Northern Hemisphere.


Genome Editing in CAR-T Cells Using CRISPR/Cas9 Technology.
CAR-T cell therapy is revolutionizing the treatment of hematologic malignancies. However, there are still many challenges ahead before CAR-T cells can be used effectively to treat solid tumors and certain hematologic cancers, such as T-cell malignancies. Next-generation CAR-T cells containing further genetic modifications are being developed to overcome some of the current limitations of this therapy. In this regard, genome editing is being explored to knock out or knock in genes with the goal of enhancing CAR-T cell efficacy or increasing access. In this chapter, we describe in detail a protocol to knock out genes on CAR-T cells using CRISPR-Cas9 technology. Among various gene editing protocols, due to its simplicity, versatility, and reduced toxicity, we focused on the electroporation of ribonucleoprotein complexes containing the Cas9 protein together with sgRNA. All together, these protocols allow for the design of the knockout strategy, CAR-T cell expansion and genome editing, and analysis of knockout efficiency.


Germ tube growth of Candida albicans.
The clinical pathogen Candida albicans is a budding yeast that is capable of forming a range of polarized and expanded cell shapes from pseudohyphae to true nonconstricted hyphae. Filamentous forms consist of contiguous uninucleated compartments that are partitioned by septa. It has long been held that the so-called "dimorphic transition" from a budding to a filamentous form may aid the fungus to penetrate epithelia and may therefore be a virulence factor. This review summarized new information regarding the physiology and ecology of hyphal growth in C. albicans. New evidence has demonstrated that hyphae of C. albicans have a sense of touch so that they grow along grooves and through pores (thigmotropism). This may aid infiltration of epithelial surfaces during tissue invasion. Hyphae are also aerotropic and can form helices when contacting solid surfaces. Growing evidence supports the view that hyphal growth is a response to nutrient deprivation, especially low nitrogen and that filamentous growth enables the fungus to forage for nutrients more effectively. Further insights into the growth of C. albicans have come from the analysis of genes and mutations of Saccharomyces which have begun to reveal the molecular mechanisms underlying the mechanisms of bud site selection, cell polarity and signal transduction pathways that lead to pseudohyphal development in this and other organisms. For example, it is now clear that a MAP-kinase cascade, homologous to the mating pathway in Saccharomyces, regulates filamentous growth in both fungi. However, this must be only one of several overlapping or separate signal transduction pathways for hyphal development because filamentous growth still occurs in mutants of Candida and Saccharomyces which are blocked in this pathway. Cell cycle analyses have shown that hyphal phase cell cycle of Candida is distinct from that in budding and pseudohyphal formation and so pseudohyphal growth of Saccharomyces is not a true model of germ tube growth in Candida. Pseudohyphal growth in both Candida and Saccharomyces involves synchronous division of mother cells and their daughters. In contrast, during germ tube growth of Candida, cytoplasm is unequally partitioned at cytokinesis so that apical cells inherit more cytoplasm and sub-apical cells have a single nucleus but are extensively vacuolated. As a result, apical cells grow and divide while sub-apical cells are apparently arrested in the cell cycle until they can regenerate sufficient cytoplasm to re-enter the cell cycle. Although current studies still fall short of verifying the status of yeast-hypha dimorphism as a virulence factor, they suggest that the cell biology of germ tube growth of C. albicans is well suited for the invasive growth of the fungus in vivo.


Biofilm Sampling for Detection of Cryptosporidium Oocysts in a Southeastern Pennsylvania Watershed.
This study investigated the use of biofilms to monitor Cryptosporidium in water. Benthic rock and submersible slide biofilms were sampled upstream and downstream of point sources in a suburban watershed in southeastern Pennsylvania. More oocysts were detected in biofilms scraped from rocks downstream than upstream of a wastewater treatment plant (WWTP) (19 versus 5, respectively; n = 1). Although not statistically significant, Cryptosporidium oocysts were detected more frequently, and in greater numbers, in biofilms grown on slides downstream than upstream of this same WWTP (83.3% positive samples [n = 12] versus 45.5% positive samples [n = 11], respectively; P = 0.0567). Similarly, Cryptosporidium oocysts were detected more frequently, and in greater numbers, in rock biofilms collected downstream than upstream of a stormwater outfall impacted by defective sewer laterals (50% positive samples downstream and 17% positive samples upstream; n = 6; P = 0.2207). While oocyst detection data obtained by slide biofilms versus filters did not necessarily agree on a given day, there was no seasonal difference in the frequency of oocyst detection (P > 0.05) or numbers of oocysts detected (P > 0.05) whether the water was monitored by filtration or slide biofilm sampling. Within any given season, there was no difference in the frequency of oocyst detection (P > 0.05) or the numbers of oocysts detected (P > 0.05) whether the water was monitored by filtration or slide biofilm sampling. These data show that oocyst detection in biofilms is comparable to oocyst detection in filtered water samples. Biofilm sampling offers significant cost savings compared to the filtration-based EPA Method 1623.1 and could be used to identify watershed locations at potential risk for increased oocyst loads.IMPORTANCE Monitoring Cryptosporidium occurrence in watersheds that provide drinking water is necessary to determine where limited resources should most effectively be directed to protect consumers from waterborne exposure to pathogenic oocysts. Biofilms are a useful tool to monitor complex watersheds and identify point sources of Cryptosporidium oocyst contamination that need to be managed to protect public health. Compared to EPA Method 1623.1, the cost benefit of using biofilms to monitor for Cryptosporidium contamination will enable utilities to sample water supplies more frequently, and at more locations, than is currently possible given limited operating budgets. Biofilm sampling could be used to identify high-risk regions within a large, complex watershed and the associated water treatment plants at potential risk for increased oocyst loads in the water supply; this information could then be used to select the locations within the watershed where the more expensive EPA Method 1623.1 is warranted.


Antagonism of IAPs Enhances CAR T-cell Efficacy.
Chimeric antigen receptor (CAR) T-cell therapy has proven successful in the treatment of hematological malignancies, notably acute lymphoblastic leukemia and B-cell lymphoma. However, the efficacy of CAR T cells against solid tumors is poor, likely due to tumor-associated immunosuppression. Here, we demonstrated that antagonizing the "inhibitor of apoptosis proteins" with the clinical smac-mimetic, birinapant, significantly enhanced the antitumor activity of CAR T cells in a tumor necrosis factor (TNF)-dependent manner. Enhanced tumor cell death occurred independently of the perforin-mediated granule exocytosis pathway, underscoring the cytotoxic potential of CAR T-cell-derived TNF. Combining CAR T-cell therapy with birinapant significantly reduced established tumor growth in vivo, where either therapy alone was relatively ineffective. Using patient biopsy-derived tumoroids, we demonstrated the synergistic potential of combining CAR T-cell therapy with smac-mimetics. Taken together, we identified CAR T-cell-derived TNF as a potent antitumor effector, which can be further harnessed by smac-mimetics.


Overweight and obesity and their relation to dietary habits and socio-demographic characteristics among male primary school children in Al-Hassa, Kingdom of Saudi Arabia.
Several studies were carried out to study the prevalence of overweight and obesity among Saudi children, but those assessed the association between eating habits, socio-demographic differentials and obesity in these children are scarce. To assess the magnitude of obesity and overweight among male primary schoolchildren and to find the possible association between obesity/overweight with dietary habits and socio-demographic differentials among them. A cross-sectional descriptive study including 1,139 Saudi male enrolled in the fifth and sixth grades in public primary schools in Al Hassa, KSA, through a multistage random sampling technique, submitted to interview using Youth and Adolescent Food Frequency Questionnaire, gathering data regarding dietary intake, some dietary habits, followed by anthropometric measurements with calculation of body mass index, the interpretation of which was based on using Cole's tables for standard definition of overweight and obesity. Socio-demographics data were collected through parental questionnaire form. Data analysis was carried out using SPSS 12 (SPSS Inc. Chicago, IL, USA), univariate as well as multivariate analyses were conducted. The age ranged from 10 to 14 years. The prevalence of overweight among the included subjects was 14.2% while obesity was 9.7%, more in urban, older age students, mothers of obese and overweight were less educated, more working. Missing and or infrequent intake of breakfast at home, frequent consumption of fast foods, low servings of fruits, vegetables, milk and dairy product per day, with frequent consumption of sweets/candy and carbonated drinks were all predictors of obesity and overweight among the included male schoolchildren. The prevalence of childhood obesity is escalating and approaching figures reported in the developed countries. Less healthy dietary habits and poor food choices may be responsible for this high prevalence.


Molecular tectonics. Porous hydrogen-bonded networks built from derivatives of pentaerythrityl tetraphenyl ether.
The symmetric four-armed geometry of pentaerythrityl tetraphenyl ether (5) makes it a valuable starting point for building complex molecular and supramolecular structures. In particular, it provides a core to which multiple sites of attractive intermolecular interaction can be attached, thereby creating compounds predisposed to form complex networks by association. To facilitate exploitation of the pentaerythrityl tetraphenyl ether core in such ways, we have prepared more than 20 new derivatives by efficient methods. Of special interest are compounds 3 and 4, which incorporate four diaminotriazine groups attached to the meta and para positions of the pentaerythrityl tetraphenyl ether core. Crystallization of compounds 3 and 4 from DMSO/dioxane is directed by hydrogen bonding of the diaminotriazine groups according to well-established motifs, thereby producing three-dimensional networks. In forming these networks, each molecule of compound 3 forms a total of 12 hydrogen bonds with six others, whereas each molecule of compound 4 forms a total of 16 hydrogen bonds with four others. Both networks are highly porous and define significant interconnected channels for the inclusion of guests. In crystals of compounds 3 and 4, the fraction of the volume accessible to guests is 66% and 57%, respectively. In both cases, the pentaerythrityl tetraphenyl ether cores adopt conformations that deviate substantially from tetrahedral geometry. It is noteworthy that the inherent flexibility of the core does not favor the formation of close-packed guest-free structures.


Levofloxacin for treatment of ventilator-associated pneumonia: a subgroup analysis from a randomized trial.
Ventilator-associated pneumonia (VAP) remains a significant challenge in critical care. We conducted a secondary analysis of a multicenter, prospective, randomized trial comparing levofloxacin (750 mg iv q24h) with imipenem-cilastatin (500-1000 mg iv q6-8h) for treatment of nosocomial pneumonia and focused on the subgroup of patients with VAP. The study cohort included 222 patients, with half (111) of the patients assigned to each treatment group. The patients in both groups were similar with respect to age, severity of illness, and duration of mechanical ventilation before the onset of VAP. Among the intention-to-treat population, clinical success was achieved in 58.6% of patients receiving levofloxacin, compared with 63.1% of patients receiving imipenem-cilastatin (P=.49; 95% confidence interval for the difference, -8.77% to 17.79%). Microbiological success and 28-day mortality rates were also comparable. Multivariate analysis demonstrated that assignment to antibiotic treatment (i.e., levofloxacin vs. imipenem-cilastatin) was not predictive of outcomes, thus suggesting that the treatment regimens were equivalent. Both levofloxacin and imipenem-cilastatin regimens were well tolerated and had similar adverse event profiles.


Multiphase CT angiography perfusion maps for predicting target mismatch and ischemic lesion volumes.
The complexity of CT perfusion (CTP) acquisition protocols may limit the availability of target mismatch assessment at resource-limited hospitals. We compared CTP mismatch with a mismatch surrogate generated from a simplified dynamic imaging sequence comprising widely available non-contrast CT (NCCT) and multiphase CT angiography (mCTA). Consecutive patients with anterior circulation acute ischemic stroke who received NCCT, mCTA, and CTP were retrospectively included in this study. An mCTA-perfusion (mCTA-P) dynamic series was formed by co-registering NCCT and mCTA. We simulated an ideal mCTA-P study by down-sampling CTP (dCTP) dynamic images according to mCTA timing. Ischemic core and penumbra volumes were estimated by cerebral blood flow and Tmax thresholding, respectively, on perfusion maps calculated independently for CTP, dCTP, and mCTA-P by deconvolution. Concordance in target mismatch (core < 70 ml, penumbra ≥ 15 ml, mismatch ratio ≥ 1.8) determination by dCTP and mCTA-P versus CTP was assessed. Of sixty-one included patients, forty-six had a CTP target mismatch. Concordance with CTP profiles was 90% and 82% for dCTP and mCTA-P, respectively. Lower mCTA-P concordance was likely from differences in collimation width between NCCT and mCTA, which worsened perfusion map quality due to a CT number shift at mCTA. Moderate diagnostic agreement between CTP and mCTA-P was found and may improve with optimal mCTA scan parameter selection as simulated by dCTP. mCTA-P may be a pragmatic alternative where CTP is unavailable or the risks of additional radiation dose, contrast injections, and treatment delays outweigh the potential benefit of a separate CTP scan.


Methanol production during chemotaxis to amino acids in Bacillus subtilis.
The 20 common amino acids act as attractants during chemotaxis by the Gram-positive organism Bacillus subtilis. In this study, we report that all amino acids induce B. subtilis to produce methanol both upon addition and removal of the chemoeffector. Asparagine-induced methanol production is specific to the McpB receptor and aspartate-induced methanol production correlates with receptor occupancy. These findings suggest that addition and removal of all amino acids cause demethylation of specific receptors which results in methanol production. We also demonstrate that certain attractants cause greater production of methanol after multiple stimulations. CheC and CheD, while affecting the levels of receptor methylation, are not absolutely required for either methylation or demethylation. In contrast, CheY is necessary for methanol formation upon removal of attractant but not upon addition of attractant. We conclude that methanol formation due to negative stimuli indicates the existence of a unique adaptational mechanism in B. subtilis involving the response regulator, CheY.


Location of the site-specific recombination system of R46: a function necessary for plasmid maintenance.
The R46 site-specific recombination system comprises a per (plasmid-encoded recombinase) gene and a site at which the gene product acts, the per site. The two functions have been cloned into pACYC184. They are encoded by sequences within a region of approximately 2 kb on the R46 genome. These R46 sequences are closely related to the site-specific recombination systems of the ampicillin resistance transposons collectively designated TnA. The R46 per function is interchangeable with the tnpR gene product of TnA. Both enzymes can mediate recombination between the related res and per sites in R46::TnA recombinant plasmids to generate site-specific deletions and inversions. Similar DNA rearrangements occur when TnA inserts into pACYC184 derivatives carrying the cloned R46 per functions. Carriage of this site-specific recombination system contributes to the stable maintenance of R46. By converting plasmid dimers to monomers the R46 per functions help to ensure equal partitioning at cell division.


Egocentric and allocentric spatial reference frames in aging: A systematic review.
Aging affects many aspects of everyday living, such as autonomy, security and quality of life. Among all, spatial memory and spatial navigation show a gradual but noticeable decline, as a result of both neurobiological changes and the general slowing down of cognitive functioning. We conducted a systematic review using the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines to identify studies that specifically investigated the role of allocentric and egocentric frames in healthy aging. Concerning spatial navigation, our results showed a preservation of egocentric strategies, along with specific impairments in the use of allocentric and switching abilities. Regarding spatial memory, instead, outcomes were more divergent and not frame-specific. With this perspective, spatial impairments were discussed considering the cognitive profile of mild cognitive impairment (MCI) and Alzheimer's Disease (AD).


Mechanism of production of pulmonary lesions in mice by Newcastle disease virus (NDV).
Infectious NDV particles produce extensive pulmonary consolidation in the mouse in the absence of demonstrable virus multiplication. The lesions are indistinguishable from those of influenza A virus infection. This effect of NDV was blocked by intranasal injection of RDE or immune serum before virus inoculation, but not by immune serum injected 5 minutes or more after NDV. Influenza A virus infection did not diminish fixation of NDV in excised lungs but did interfere with the injurious action of this agent in the living mouse. The analogy between these reactions and those which take place in a progressive virus infection is pointed out, and the mechanism of production of lesions in virus pneumonias discussed.


Dementia syndromes: neurobehavioral and neuropsychiatric features.
Dementia is seen in a wide variety of conditions that, in general, lack specific laboratory features. Etiologic classification depends on a combination of historical/clinical and radiologic/EEG considerations. Neurobehavioral and neuropsychiatric characteristics of the dementia syndromes may be key to accurate diagnosis. The diseases that produce dementia syndromes, including degenerative, vascular, myelinoclastic, traumatic, infective, and metabolic disorders, are reviewed, and the principles that permit differential diagnosis are discussed.


Current and future management options for myelodysplastic syndromes.
The management of the myelodysplastic syndromes (MDS) requires insight into the complex biology of the disease. Despite this challenge, two recent developments have contributed significantly to advancements in the treatment of MDS: (i) improvements in classification systems and prognostic models; and (ii) the emergence of US FDA-approved agents such as lenalidomide, azacitidine and decitabine. Prior to these developments, supportive care measures consisting of blood and platelet transfusions, haematopoietic growth factors and antimicrobials remained standard of care for the treatment of MDS. As a result of these developments, clinicians are able to provide patient-tailored therapy for specific MDS subgroups. Clinical trials addressing combination therapies with multiple investigational agents as well as novel combination regimens are ongoing. This review focuses on supportive care modalities, the approved agents indicated for the treatment of MDS and future directions for the treatment of MDS, including agents under clinical investigation.


Honeybush Extracts (Cyclopia spp.) Rescue Mitochondrial Functions and Bioenergetics against Oxidative Injury.
Mitochondrial dysfunction plays a major role not only in the pathogenesis of many oxidative stress or age-related diseases such as neurodegenerative as well as mental disorders but also in normal aging. There is evidence that oxidative stress and mitochondrial dysfunction are the most upstream and common events in the pathomechanisms of neurodegeneration. Cyclopia species are endemic South African plants and some have a long tradition of use as herbal tea, known as honeybush tea. Extracts of the tea are gaining more scientific attention due to their phenolic composition. In the present study, we tested not only the in vitro mitochondria-enhancing properties of honeybush extracts under physiological conditions but also their ameliorative properties under oxidative stress situations. Hot water and ethanolic extracts of C. subternata, C. genistoides, and C. longifolia were investigated. Pretreatment of human neuroblastoma SH-SY5Y cells with honeybush extracts, at a concentration range of 0.1-1 ng/ml, had a beneficial effect on bioenergetics as it increased ATP production, respiration, and mitochondrial membrane potential (MMP) after 24 hours under physiological conditions. The aqueous extracts of C. subternata and C. genistoides, in particular, showed a protective effect by rescuing the bioenergetic and mitochondrial deficits under oxidative stress conditions (400 μM H2O2 for 3 hours). These findings indicate that honeybush extracts could constitute candidates for the prevention of oxidative stress with an impact on aging processes and age-related neurodegenerative disorders potentially leading to the development of a condition-specific nutraceutical.


Love, Work, and Striving for the Self in Balance: Anaclitic and Introjective Patients' Experiences of Change in Psychoanalysis.
One of the most famous quotations credited to Freud is that, when asked what he thought a psychologically healthy person should be able to do, he said: "to love and to work." A central goal in psychoanalytic treatment is to bring about changes in basic, mostly unconscious, mental structures. The aim of this study was to investigate, applying an inductive thematic analysis, the experiences anaclitic and introjective patients have had of change after psychoanalysis with regard to the domains Love and Relationships and Work and Achievements. Analyzing patient interviews, we identified a third domain of experienced changes, The Self, which refers to increased self-understanding, self-acceptance, and self-care rather than an improved dynamic balance between love and work. All patients experienced several positive changes in their lives during and after psychoanalysis. We also found distinctive patterns that appear to be closely linked to the patients' initial personality orientation with regard to relationships and achievements. Generally, the patients described symmetrical, but opposite, change processes within the two specific domains of Love and Work. For the anaclitic patients, this indicated a movement inward in the domain of Love (from an excessive preoccupation with issues of their relationship with others toward more distinct self-boundaries and increased agency) and outward in the domain of Work (from unenterprising toward becoming more outgoing and daring). For the introjective patients, this pointed to a reverse movement outward in the domain of Love (from an excessive preoccupation with issues of autonomy toward increased responsiveness to others and desire to be establish close, mutual relationships) and inward in the domain of Work (from an excessive orientation on achievements toward increased becoming more grounded in their own feelings, needs, and desires). In conclusion, patients in both groups have experienced a reduced preoccupation with issues related to their initially predominant personality dimension (relatedness or self-definition) and increased receptivity to needs typical for the complementary dimension. These changes seem to be mediated by changes in the domain of The Self. Our study suggests the clinical relevance of focusing the therapeutic work on fostering a better and more dynamic balance between love and work, relatedness, or self-definition.


Classifying Chronic Lower Respiratory Disease Events in Epidemiologic Cohort Studies.
One in 12 adults has chronic obstructive pulmonary disease or asthma. Acute exacerbations of these chronic lower respiratory diseases (CLRDs) are a major cause of morbidity and mortality. Valid approaches to classifying cases and exacerbations in the general population are needed to facilitate prevention research. To assess the feasibility, reproducibility, and performance of a protocol to identify CLRD cases and exacerbations triggering emergency department (ED) visits or hospitalizations in cohorts of patients derived from general populations of adults. A protocol was developed to classify CLRD cases and severe exacerbations on the basis of review of medical records. ED and inpatient medical records were ascertained prospectively in the Hispanic Community Health Study/Study of Latinos, and inpatient records were retrospectively identified by administrative codes in the Multi-Ethnic Study of Atherosclerosis. "Probable" exacerbations were defined as a physician's diagnosis of CLRD with acute respiratory symptoms. "Highly probable" exacerbations additionally required systemic corticosteroid therapy, and "definite" exacerbations required airflow limitation or evidence of CLRD on imaging studies. Adjudicated results were compared with CLRD cases identified by spirometry and self-report, and with an administrative definition of exacerbations. Protocol-based classification was completed independently by two physicians for 216 medical records (56 ED visits and 61 hospitalizations in the Hispanic Community Health Study/Study of Latinos; 99 hospitalizations in the Multi-Ethnic Study of Atherosclerosis). Reviewer disagreement occurred in 2-5% of cases and 4-8% of exacerbations. Eighty-nine percent of records were confirmed as at least probable CLRD cases. Fifty-six percent of confirmed CLRD cases had airflow limitation on the basis of baseline study spirometry. Of records that described CLRD as the primary discharge diagnosis code, an acute exacerbation was confirmed as at least probable for 96% and as highly probable or definite for 77%. Only 50% of records with CLRD as a secondary code were confirmed, although such records accounted for over half of all confirmed exacerbations. CLRD cases and severe exacerbations without preceding documentation of airflow limitation are identified frequently in population-based cohorts of persons. A primary discharge diagnosis of CLRD is specific but insensitive for defining exacerbations. Protocol-based classification of medical records may be appropriate to supplement and to validate identification of CLRD cases and exacerbations in general population studies. Clinical trials registered with www.clinicaltrials.gov (NCT00005487 and NCT02060344).


Interspecific transplantation of polar plasm between Drosophila embryos.
Posterior polar plasm of the Drosophila egg has been shown to function autonomously in germ cell determination after transplantation to either the anterior or mid-ventral region of the early embryo. By means of similar transplantations, we have tested the ability of polar plasm of Drosophila immigrans to induce the formation of pole cells in a Drosophila melanogaster embryo. After the transplantation of polar plasm, "hybrid" pole cells were found in which both pole cell-specific organelles, the polar granules and nuclear body, were structurally similar to those characteristic of the transplanted cytoplasm. In order to determine whether these hybrid cells can function as germ cell precursors, these cells were transplanted to the posterior tip of genetically marked embryos. Approximately 5% of the flies obtained from embryos receiving potential pole cells produce offspring derived from the induced pole cells. This result demonstrates that polar plasm can function in interspecific species combinations and indicates that the molecular mechanisms of germ cell determination are conservative in evolution. Finally, in order to test whether there is any evidence for cytoplasmic inheritance of polar granules, embryos derived from hybrid pole cells were examined for their polar granule morphology. The fine structure of the granules conformed to that of the nucleus. Thus, no evidence was found for the cytoplasmic inheritance of these particular organelles.


Feeder-free culture of human embryonic stem cells in conditioned medium for efficient genetic modification.
Realizing the potential of human embryonic stem cells (hESCs) in research and commercial applications requires generic protocols for culture, expansion and genetic modification that function between multiple lines. Here we describe a feeder-free hESC culture protocol that was tested in 13 independent hESC lines derived in five different laboratories. The procedure is based on Matrigel adaptation in mouse embryonic fibroblast conditioned medium (CM) followed by monolayer culture of hESC. When combined, these techniques provide a robust hESC culture platform, suitable for high-efficiency genetic modification via plasmid transfection (using lipofection or electroporation), siRNA knockdown and viral transduction. In contrast to other available protocols, it does not require optimization for individual lines. hESC transiently expressing ectopic genes are obtained within 9 d and stable transgenic lines within 3 weeks.


Hand-grip strength predicts incident disability in non-disabled older men.
To verify if hand-grip performance in older men is a predictor of disability. Population-based prospective study. A sample from the Italian rural cohorts of the FINE study (Finland, Italy, Netherlands Elderly), representative of the general population of elderly men surveyed in 1991 and 1995. 140 men aged 71-91 years who reported no disability in performing activities of daily living (ADLs), instrumental activity of daily living (IADLs) and mobility activities at baseline examination and provided information on their functional status at follow-up 4 years later. Disability was defined as needing help in performing ADLs, IADLs and mobility. Hand-grip strength was evaluated at baseline by a mechanical dynamometer. After adjusting for potential confounding variables, a lower concentration of high-density lipoprotein cholesterol was the only factor predicting disability in men aged 76 years or younger and only reduced hand-grip strength predicted incident disability in men 77 years or older. Poor hand strength as measured by hand-grip is a predictor of disability in older people. The hand-grip test is an easy and inexpensive screening tool to identify elderly people at risk of disability.


Property modelling of lysozyme-crosslinker-alginate complexes using latent variable methods.
Statistical methods were used to provide insight into a polymer complex system composed of lysozyme and alginate to quantify the effects of such parameters as pH, and ionic composition of the mixing solution on the properties of the complexes including composition, particle diameter, and zeta potential. Various crosslinkers (calcium, barium, iron[III], and bovine serum albumin), were used with lysozyme for complex formation to investigate the effect of crosslinker charge density on protein release kinetics, modelled using ktn . Multivariate statistical analysis showed that the kinetic parameters associated with the release were, not surprisingly highly dependent on the ionic strength of the release media, with higher ionic strength leading to faster release. The release parameter k was also shown to depend on the protein properties (size, ionic strength) while n was slightly, but not statistically dependent on the charge density of the crosslinker demonstrating that the nature of the crosslinker had minimal impact on drug release. The multivariate statistical has the potential to be used for optimization of the complexes and prediction of physical properties and degradation rates.


The cost-effectiveness of screening for colorectal cancer.
Published decision analyses show that screening for colorectal cancer is cost-effective. However, because of the number of tests available, the optimal screening strategy in Canada is unknown. We estimated the incremental cost-effectiveness of 10 strategies for colorectal cancer screening, as well as no screening, incorporating quality of life, noncompliance and data on the costs and benefits of chemotherapy. We used a probabilistic Markov model to estimate the costs and quality-adjusted life expectancy of 50-year-old average-risk Canadians without screening and with screening by each test. We populated the model with data from the published literature. We calculated costs from the perspective of a third-party payer, with inflation to 2007 Canadian dollars. Of the 10 strategies considered, we focused on three tests currently being used for population screening in some Canadian provinces: low-sensitivity guaiac fecal occult blood test, performed annually; fecal immunochemical test, performed annually; and colonoscopy, performed every 10 years. These strategies reduced the incidence of colorectal cancer by 44%, 65% and 81%, and mortality by 55%, 74% and 83%, respectively, compared with no screening. These strategies generated incremental cost-effectiveness ratios of $9159, $611 and $6133 per quality-adjusted life year, respectively. The findings were robust to probabilistic sensitivity analysis. Colonoscopy every 10 years yielded the greatest net health benefit. Screening for colorectal cancer is cost-effective over conventional levels of willingness to pay. Annual high-sensitivity fecal occult blood testing, such as a fecal immunochemical test, or colonoscopy every 10 years offer the best value for the money in Canada.


COVID-19 and the burning issue of drug interaction: never forget the ECG.
The coronavirus disease of 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV2), has been rapidly escalating, becoming a relevant threat to global health. Being a recent virus outbreak, there are still no available therapeutic regimens that have been approved in large randomised trials and so patients are currently being treated with multiple drugs. This raises concerns regarding drug interaction and their implication in arrhythmic burden. In fact, two of the actually used drugs against SARS-CoV2, such as chloroquine and the combination lopinavir/ritonavir, might determine a QT (the time from the start of the Q wave to the end of the T wave) interval prolongation and they show several interactions with antiarrhythmic drugs and antipsychotic medications, making them prone to an increased risk of developing arrhythmias. This brief review focuses the attention on the most relevant drug interactions involving the currently used COVID-19 medications and their possible association with cardiac rhythm disorders, taking into account also pre-existing condition and precipitating factors that might additionally increase this risk. Furthermore, based on the available evidence and based on the knowledge of drug interaction, we propose a quick and simple algorithm that might help both cardiologists and non-cardiologists in the management of the arrhythmic risk before and during the treatment with the specific drugs used against SARS-CoV2.


[Molecular genetic analysis of a diversity of Getah virus strains isolated from mosquitoes in the North-Eastern Asia].
A molecular genetic study was performed of the Getah virus strains isolated in Russia (Eastern Siberia and Far East) and Mongolia. A phylogenetic analysis was made, by examining the nucleotide sequences of genome fragments that had been obtained by RT-PCR and that included the portions of the E1 and E2 surface glycoprotein genes and the 6K gene. A genetic diversity of Getah virus strains in North-Eastern Asia is discussed.


Altered hepatic lipid droplet morphology and lipid metabolism in fasted Plin2-null mice.
Perilipin 2 (Plin2) binds to the surface of hepatic lipid droplets (LDs) with expression levels that correlate with triacylglyceride (TAG) content. We investigated if Plin2 is important for hepatic LD storage in fasted or high-fat diet-induced obese Plin2+/+ and Plin2-/- mice. Plin2-/- mice had comparable body weights, metabolic phenotype, glucose tolerance, and circulating TAG and total cholesterol levels compared with Plin2+/+ mice, regardless of the dietary regime. Both fasted and high-fat fed Plin2-/- mice stored reduced levels of hepatic TAG compared with Plin2+/+ mice. Fasted Plin2-/- mice stored fewer but larger hepatic LDs compared with Plin2+/+ mice. Detailed hepatic lipid analysis showed substantial reductions in accumulated TAG species in fasted Plin2-/- mice compared with Plin2+/+ mice, whereas cholesteryl esters and phosphatidylcholines were increased. RNA-Seq revealed minor differences in hepatic gene expression between fed Plin2+/+ and Plin2-/- mice, in contrast to marked differences in gene expression between fasted Plin2+/+ and Plin2-/- mice. Our findings demonstrate that Plin2 is required to regulate hepatic LD size and storage of neutral lipid species in the fasted state, while its role in obesity-induced steatosis is less clear.


A target tracking method based on adaptive occlusion judgment and model updating strategy.
Target tracking is an important research in the field of computer vision. Despite the rapid development of technology, difficulties still remain in balancing the overall performance for target occlusion, motion blur, etc. To address the above issue, we propose an improved kernel correlation filter tracking algorithm with adaptive occlusion judgement and model updating strategy (called Aojmus) to achieve robust target tracking. Firstly, the algorithm fuses color-naming (CN) and histogram of gradients (HOG) features as a feature extraction scheme and introduces a scale filter to estimate the target scale, which reduces tracking error caused by the variations of target features and scales. Secondly, the Aojmus introduces four evaluation indicators and a double thresholding mechanism to determine whether the target is occluded and the degree of occlusion respectively. The four evaluation results are weighted and fused to a final value. Finally, the updating strategy of the model is adaptively adjusted based on the weighted fusion value and the result of the scale estimation. Experimental evaluations on the OTB-2015 dataset are conducted to compare the performance of the Aojmus algorithm with four other comparable algorithms in terms of tracking precision, success rate, and speed. The experimental results show that the proposed Aojmus algorithm outperforms all the algorithms compared in terms of tracking precision. The Aojmus also exhibits excellent performance on attributes such as target occlusion and motion blur in terms of success rate. In addition, the processing speed reaches 74.85 fps, which also demonstrates good real-time performance.


DNA binding by yeast Mlh1 and Pms1: implications for DNA mismatch repair.
The yeast Mlh1-Pms1 heterodimer required for mismatch repair (MMR) binds to DNA. Here we map DNA binding to N-terminal fragments of Mlh1 and Pms1. We demonstrate that Mlh1 and Pms1 N-terminal domains (NTDs) independently bind to double-stranded and single-stranded DNA, in the absence of dimerization and with different affinities. Full-length Mlh1p alone, which can homodimerize, also binds to DNA. Substituting conserved positively charged amino acids in Mlh1 produces mutator phenotypes in a haploid yeast strain characteristic of reduced MMR. These substitutions strongly reduce DNA binding by the Mlh1 NTD and, to a lesser extent, they also reduce DNA binding by full-length Mlh1 and the Mlh1-Pms1 heterodimer. Replacement of a homologous Pms1 residue has a much smaller effect on mutation rate and does not reduce DNA binding. The results demonstrate that NTDs of yeast Mlh1 and Pms1 contain independent DNA binding sites and they suggest that the C-terminal region of Mlh1p may also contribute to DNA binding. The differential mutator effects and binding properties observed here further suggest that Mlh1 and Pms1 differ in their interactions with DNA. Finally, the results are consistent with the hypothesis that DNA binding by Mlh1 is important for MMR.


Silencing Hsp25/Hsp27 gene expression augments proteasome activity and increases CD8+ T-cell-mediated tumor killing and memory responses.
Relatively high expression of Hsp27 in breast and prostate cancer is a predictor of poor clinical outcome. This study elucidates a hitherto unknown mechanism by which Hsp27 regulates proteasome function and modulates tumor-specific T-cell responses. Here, we showed that short-term silencing of Hsp25 or Hsp27 using siRNA or permanent silencing of Hsp25 using lentivirus RNA interference technology enhanced PA28α mRNA expression, PA28α protein expression, and proteasome activity; abrogated metastatic potential; induced the regression of established breast tumors by tumor-specific CD8(+) T cells; and stimulated long-lasting memory responses. The adoptive transfer of reactive CD8(+) T cells from mice bearing Hsp25-silenced tumors efficiently induced the regression of established tumors in nontreated mice which normally succumb to tumor burden. The overexpression of Hsp25 and Hsp27 resulted in the repression of normal proteasome function, induced poor antigen presentation, and resulted in increased tumor burden. Taken together, this study establishes a paradigm shift in our understanding of the role of Hsp27 in the regulation of proteasome function and tumor-specific T-cell responses and paves the way for the development of molecular targets to enhance proteasome function and concomitantly inhibit Hsp27 expression in tumors for therapeutic gain.


An Explanation for Reports of Increased Prevalence of Olfactory Dysfunction with Omicron: Asymptomatic Infections.
The prevalence of olfactory dysfunction (OD) in people infected with the omicron variant is substantially reduced compared with previous variants. However, four recent studies reported a greatly increased prevalence of OD with omicron. We provide a likely explanation for these outlier studies and reveal a major methodological flaw. When the proportion of asymptomatic infections is large, then studies on the prevalence of OD will examine and report predominantly on non-representative cohorts, those with symptomatic subjects, thereby artificially inflating OD prevalence by up to ten-fold. Estimation of the true OD prevalence requires representative cohorts that include relevant fractions of asymptomatic cases.


Population pharmacokinetics of (R)-albuterol and (S)-albuterol in pediatric patients aged 4-11 years with asthma.
To characterize the population pharmacokinetics (PK) of (R)- and (S)-albuterol in pediatric asthmatics using a model that supports a sparse blood sampling strategy. The data for this analysis were collected from patients enrolled in a randomized, double-blind, multicenter, placebo- and active-controlled study evaluating the safety and efficacy of levalbuterol in asthmatic children aged 4-11 years. Patients received either levalbuterol 0.31 mg, levalbuterol 0.63 mg, racemic albuterol 1.25 mg, or racemic albuterol 2.5 mg via nebulizer. Separate population pharmacokinetic models were developed for (R)- and (S)-albuterol using the NOMNEM((R)) computer program. Covariate models were developed to identify significant predictors of inter-patient variability. A total of 995 samples and 262 patients were used for the (R)-albuterol population PK model while a total of 496 samples and 128 patients were used for the (S)-albuterol population PK model. The apparent clearance of (R)-albuterol was much more rapid than that of (S)-albuterol (approximately four-fold higher), and the apparent volume of distribution was much larger for (R)-albuterol (in part due to pre-systemic metabolism) than for (S)-albuterol (approximately four-fold higher). In this study of pediatric patients, the models were able to demonstrate using two to four samples per patient that the apparent clearance and volume of distribution of (R)-albuterol were several fold higher than that of (S)-albuterol. The pharmacokinetics of (R)-albuterol were similar after administration of levalbuterol or racemic albuterol and were linear over the examined dose range (0.31-0.63 mg nebulized dose). The presence of (S)-albuterol did not significantly alter the pharmacokinetics of (R)-albuterol, suggesting that effects of (S)-albuterol may be due to the intrinsic pharmacology of this isomer.


The protective effect of supplemental calcium on colonic permeability depends on a calcium phosphate-induced increase in luminal buffering capacity.
An increased intestinal permeability is associated with several diseases. Previously, we have shown that dietary Ca decreases colonic permeability in rats. This might be explained by a calcium-phosphate-induced increase in luminal buffering capacity, which protects against an acidic pH due to microbial fermentation. Therefore, we investigated whether dietary phosphate is a co-player in the effect of Ca on permeability. Rats were fed a humanised low-Ca diet, or a similar diet supplemented with Ca and containing either high, medium or low phosphate concentrations. Chromium-EDTA was added as an inert dietary intestinal permeability marker. After dietary adaptation, short-chain fructo-oligosaccharides (scFOS) were added to all diets to stimulate fermentation, acidify the colonic contents and induce an increase in permeability. Dietary Ca prevented the scFOS-induced increase in intestinal permeability in rats fed medium- and high-phosphate diets but not in those fed the low-phosphate diet. This was associated with higher faecal water cytotoxicity and higher caecal lactate levels in the latter group. Moreover, food intake and body weight during scFOS supplementation were adversely affected by the low-phosphate diet. Importantly, luminal buffering capacity was higher in rats fed the medium- and high-phosphate diets compared with those fed the low-phosphate diet. The protective effect of dietary Ca on intestinal permeability is impaired if dietary phosphate is low. This is associated with a calcium phosphate-induced increase in luminal buffering capacity. Dragging phosphate into the colon and thereby increasing the colonic phosphate concentration is at least part of the mechanism behind the protective effect of Ca on intestinal permeability.


[Cerebral hemodynamics in neurocirculatory hypotension].
Data are presented on the state of cerebral haemodynamics in patients with neuro-circulatory hypotension that was studied by measuring the arterial pressure in various zones of blood supply (the central retinal artery, superficial temporal artery and the brachial artery), by studying the ocular fundus and by rheoencephalography. The cerebral haemodynamic disorders were revealed in the form of regional cerebral hypertension in 13.4% and regional cerebral hypotension in 29.4%, against the background of a general fall of the arterial pressure. In regional cerebral hypertension the vessels of the ocular fundus were narrowed, in regional cerebral hypotension--vice versa. The reduction of the pulse pressure during neuro-circulatory hypotension also attracts attention, and it can be considered a sign of impaired blood supply to the brain.


Clinical Evaluation for the Role of High-Sensitivity C-Reactive Protein in Combination with D-Dimer and Wells Score Probability Test to Predict the Incidence of Deep Vein Thrombosis Among Cancer Patients.
Deep vein thrombosis (DVT) is a frequent complication in cancer patients and is the second leading cause of death. The high level of C-reactive protein (CRP) is an acute phase reactant that induces tissue factor (TF) expression in monocytes, smooth muscle cells, and endothelial cells. The CRP level positively correlates with the incidence, extension, and volume of thrombus. TF expression triggers the coagulation system including the formation of thrombin and circulating fibrin such as prothrombin fragment 1+2 (F1 + 2) and D-dimer. To determine the diagnostic value of high-sensitivity (hs)-CRP, D-dimer, and Wells score combination to predict the incidence of DVT on clinically suspected DVT (Wells score ≥2) cancer patients. This study was a cross-sectional study on a diagnostic test to determine the diagnostic value of hs-CRP and D-dimer for early detection of DVT on clinically suspected DVT (Wells score ≥2) cancer patients. It was conducted in Dr. Kariadi Hospital, Semarang Indonesia on 35 subjects. The diagnosis of DVT was confirmed by color duplex sonography. The diagnostic accuracy of combination of hs-CRP, D-dimer, and Wells score was analyzed by logistic regression. DVT was confirmed in 10 subjects (28,6%). The cut-off point of hs-CRP levels for probable DVT was ≥51.05 mg/L and for D-dimer was ≥5030 µg/L. The median levels of both variables were higher in the subjects with DVT compared with the subjects without DVT, but it was not statistically significant. The combination of hs-CRP (≥51.05 mg/L), D-dimer (≥5030 µg/L), and Wells score ≥3 had the high accuracy (94.1%) to predict the incidence of DVT compared with hs-CRP (65.0%), D-dimer (54.7%), and combination of hs-CRP and D-dimer (71.0%). The combination of hs-CRP (≥51.05 mg/L), D-dimer (≥5030 µg/L), and Wells score ≥3 can predict the incidence of DVT in cancer.


Role of hysterectomy in the management of patients with gestational trophoblastic neoplasia: importance of receiving treatment in reference centers.
To evaluate hysterectomy prevalence, indications and impact on clinical outcomes in a reference center in southern Brazil. Institutional Ethical Committee approval was granted for this study. In a cohort study spanning 21 years, all patients who underwent hysterectomy for gestational trophoblastic neoplasia (GTN) were included, and technical differences between hysterectomy performed in the reference center and those performed elsewhere were evaluated as well. Of 1,023 patients with gestational trophoblastic disease, 57 (5.6%) underwent hysterectomy (95% CI, 4.3-7.1). Hysterectomy incidence in 230 GTN patients was 17.7% (95%CI, 15.1-23.3). Indications for 41 hysterectomies in the reference center were as follows: primary treatment in 14 (34.1%) cases and secondary treatment in 27 (65.9%); of these, the main indications were GTN recurrence (7 [25.9%] cases), hemorrhage (6 [22.2%]), resistance to single-agent chemotherapy in patients who refused more aggressive treatment (6 [22.2%]), and tumor mass reduction (5 [18.5%]). Twelve (92.3%) of the 13 hysterectomies with bilateral oophorectomy were performed elsewhere (p < 0.001). Thirty-five (85.4%) patients had no complications, and median hospitalization time was short (3 +/- 4 days). None of the 4 deaths were associated with hysterectomy. In the reference center, when associated with hysterectomy, GTN cure rates reached 93% after 63 +/- 87 months of follow-up. When treatment is in a reference center, hysterectomy frequency and morbidity may be low, and indications due to hemorrhage are significantly lower. Furthermore, at a reference center there is significantly greater ovarian preservation at the time of hysterectomy, and significantly more patients who undergo hysterectomy have low-risk GTN.


Cyclists and triathletes have different body positions on the bicycle.
Our study evaluated differences in body position on the bicycle for recreational cyclists, competitive cyclists and triathletes. Thirty-six recreational cyclists, 17 competitive road cyclists and 18 competitive triathletes were assessed for body position on their bicycles on a cycle trainer. Images were taken of cyclists/triathletes in static poses with the crank at the 3 o'clock and 6 o'clock positions. Trunk, pelvis, hip, knee and ankle angles, anterior-posterior and medio-lateral positions of the knees in relation to the pedal axis and frontal projected area were measured using ImageJ. Comparison of body position between groups (recreational, competitive road cyclists and competitive triathletes) was conducted using effects sizes (ES). The greatest differences between groups in the measured variables were observed between the triathletes and the other two groups. Smaller differences were observed between competitive and recreational cyclists. Competitive triathletes had greater body forward projection (10% greater trunk flexion and 66% knee anterior position, ES = 2.5 and 1.2, respectively) and less frontal projected area (17%, ES = 1.3) than competitive road cyclists for body position on the bicycle. Both recreational and competitive cyclists sat on their bicycles with their trunks in a more vertical position compared to triathletes. Guidelines for bicycle configuration for triathletes and road cyclists need to consider the body positions during events.


Associations between risk of overall mortality, cause-specific mortality and level of inflammatory factors with extremely low and high high-density lipoprotein cholesterol levels among American adults.
The health outcomes associated with extremely low or high plasma concentrations of high-density lipoprotein cholesterol (HDL-C) are not well documented mainly because of the small numbers of participants with such values included in the clinical trials. We prospectively investigated the association between extremely low and high HDL-C with: 1) the risk of overall, coronary heart disease (CHD), cerebrovascular and cancer mortality, and, 2) their link with inflammatory factors. Analysis was based on subjects ≥18 years old from the National Health and Nutrition Examination Surveys (NHANES). We categorized HDL-C levels as follows: [low HDL-C group ≤30 (extremely low), 30-40 (low), and ≥40 (reference)] [high HDL-C group = 40-80 (reference), 80-100 (high) and ≥100 mg/dl (extremely high). Cox proportional hazard regression models and analysis of covariance accounted for survey design, masked variance and sample weights. After adjustment for age, race and sex, we found that the very low HDL-C category (<30 mg/dl) had a greater risk of total mortality (risk ratio [RR]: 3.00, 95%CI: 2.20-4.09). RR for CHD and stroke mortality was 2.00 and 2.53, respectively; there was no link between cancer and level of HDL-C (p = 0.235). The association between total mortality, CHD and stroke with the level of HDL-C attenuated but remained significant even after adjustment for demographics, dietary, cardiovascular risk factors and treatment for dyslipidemia (all p < 0.001). After adjustments, subjects with extremely high HDL-C levels had a higher risk of mortalities (all p < 0.001). Mexican-American ethnicity, subjects in the low level of HDL-C (30-40 mg/dl) category had higher risk of mortalities than those with a very low level (all p < 0.001). Concentration of C-reactive protein, fibrinogen and white blood count significantly decreased as the level of the HDL-C increased; these findings were robust after adjustment for demographics, dietary, cardiovascular risk factors and treatment for dyslipidemia (all p < 0.001); further subjects with extremely high HDL-C levels have a greater levels of inflammatory factors (all p < 0.001). Both extremely low and high HDL-C levels were associated with greater risk of mortalities (total, CHD and stroke) and higher level of inflammatory factors, while there was no link between level of HDL-C and risk of cancer. Moreover, we found evidence of an HDL-C paradox in Mexican-American ethnicity participants.


ATP from synaptic terminals and astrocytes regulates NMDA receptors and synaptic plasticity through PSD-95 multi-protein complex.
Recent studies highlighted the importance of astrocyte-secreted molecules, such as ATP, for the slow modulation of synaptic transmission in central neurones. Biophysical mechanisms underlying the impact of gliotransmitters on the strength of individual synapse remain, however, unclear. Here we show that purinergic P2X receptors can bring significant contribution to the signalling in the individual synaptic boutons. ATP released from astrocytes facilitates a recruitment of P2X receptors into excitatory synapses by Ca(2+)-dependent mechanism. P2X receptors, co-localized with NMDA receptors in the excitatory synapses, can be activated by ATP co-released with glutamate from pre-synaptic terminals and by glia-derived ATP. An activation of P2X receptors in turn leads to down-regulation of postsynaptic NMDA receptors via Ca(2+)-dependent de-phosphorylation and interaction with PSD-95 multi-protein complex. Genetic deletion of the PSD-95 or P2X4 receptors obliterated ATP-mediated down-regulation of NMDA receptors. Impairment of purinergic modulation of NMDA receptors in the PSD-95 mutants dramatically decreased the threshold of LTP induction and increased the net magnitude of LTP. Our findings show that synergistic action of glia- and neurone-derived ATP can pre-modulate efficacy of excitatory synapses and thereby can have an important role in the glia-neuron communications and brain meta-plasticity.


Statistical study and experimental verification of high-resolution methods in phase-shifting interferometry.
The introduction of high-resolution phase-shifting interferometry methods such as annihilation filter, state space, multiple-signal classification, minimum norm, estimation of signal parameter via rotational invariance, and maximum-likelihood estimator have enabled the estimation of phase in an interferogram in the presence of harmonics and noise. These methods are also effective in holographic moiré where incorporating two piezoelectric transducers (PZTs) yields two orthogonal displacement components simultaneously. Typically, when these methods are used, the first step involves estimating the phase steps pixelwise; then the interference phase distribution is computed by designing a Vandermonde system of equations. In this context, we present a statistical study of these methods for the case of single and dual PZTs. The performance of these methods is also compared with other conventional benchmarking algorithms involving the single PZT. The paper also discusses the significant issue of an allowable pair of phase steps in the presence of noise using a robust statistical tool such as the Cramér-Rao bound. Furthermore, experimental validations of these high-resolution methods are presented for the estimation of single phase in holographic interferometry and for the estimation of multiple phases in holographic moiré.


Alternative Molecular Methods for Improved Detection of Meningococcal Carriage and Measurement of Bacterial Density.
Conventional methods for detecting pharyngeal carriage of Neisseria meningitidis are complex. There is a need for simpler methods with improved performance. We have investigated two alternative approaches. Three pharyngeal swabs were collected from 999 pupils aged 10 to 18 years in The Gambia. Carriage of N. meningitidis was investigated by using three different methods: (i) plating on Thayer-Martin selective medium and testing by conventional microbiological methods followed by PCR testing; (ii) seeding in Todd-Hewitt broth (THB) and, after culture overnight, testing by PCR; and (iii) compression of the swab on filter paper and, after DNA concentration, testing by PCR. PCR after culture in THB was more than twice as sensitive as conventional methods in detecting N. meningitidis (13.2% versus 5.7%; P < 0.0001). PCR after DNA extraction from filter paper had a sensitivity similar to that of conventional methods (4.9% versus 5.7%; P = 0.33). Capsular genogroups detected by broth culture were genogroups W (21 isolates), B (12 isolates), Y (8 isolates), E (3 isolates), and X (2 isolates), and 68 meningococci had the capsule-null intergenic region. The distributions of genogroups and of capsule-null organisms were similar with each of the three methods. The carriage density in samples extracted from filter paper ranged from 1 to 25,000 DNA copies. PCR of broth cultures grown overnight doubled the yield of N. meningitidis carriage isolates compared with conventional methods. This approach could improve the efficiency of carriage studies. Collection on filter paper followed by quantitative PCR could be useful for density measurement and for carriage studies in areas with limited resources.


Geothermal Heat Flow in the Gulfs of California and Aden.
Eighteen measurements in and near the gulfs of California and Aden indicate the geothermal flux is several times the world-wide mean of 1.2 x 10(-6) cal/cm(2) sec in both regions. Both gulfs closely coincide with the intersection of oceanic rises with continents and have likely been formed under tensional forces, which suggests an association with mantle convection currents.


Flexible employment and nurses' intention to leave the profession: The role of support at work.
The objectives of this paper are to examine (1) the association between flexible employment and nurses' intention to leave the profession, and (2) whether or not support at work mediates the association between flexible employment and nurses' intention to leave the profession. Flexible employment is analyzed objectively using non-permanent contract, part-time employment status, casual employment status, involuntary hours and on-call work, and subjectively using job insecurity. Support at work refers to organizational, supervisor and peer support. Data come from our survey of 1396 nurses employed in three teaching hospitals in Southern Ontario. Descriptive statistics are provided. Bivariate correlations, hierarchical regression analysis and mediation tests are conducted. Compared to those in full-time employment, nurses in part-time employment do not intend to leave the profession. None of the other objective flexible employment factors are associated with intention to leave the profession. Perceived job insecurity is associated with intention to leave the profession. Low support at work contributes to intention to leave the profession and mediates the association between job insecurity and intention to leave the profession. The study provides evidence to health sector managers and policy makers that part-time employment, perceived job security and support at work are important factors to consider in efforts to retain nurses in the profession.


Failure to acknowledge high suicide risk among veterinarians.
A high suicide risk has been reported among veterinarians in comparison to the general population. Postulated causes have included depression, substance abuse, work-related stress, reluctance to admit psychiatric problems, and access to lethal drugs and/or familiarity with euthanasia. Members of the Student Chapter of the American Veterinary Medical Association (AVMA), all veterinarians licensed in Alabama, and all US veterinary-association executive directors were surveyed regarding their attitudes concerning mental health issues, including veterinarian suicide. Only 10% of veterinary student respondents (N=58) believed that suicide risk is higher among veterinarians than in the general population. Of the 22 state associations' executive directors who participated in the survey, 37% believed that suicide is a significant concern for veterinarians and only 44% indicated that a veterinary wellness program was available in their respective states. Of the 1,455 licensed veterinarians in Alabama, 701 responded to the survey; 11% of respondents believed that suicide among veterinarians was a problem. In addition, 66% of respondents indicated that they had been "clinically depressed," but 32% of those with depression had not sought treatment. More females (27%) than males (20%) admitted that they had "seriously considered suicide" (p<.01). Female veterinarians were more likely than male veterinarians (15% versus 7%) to indicate that they were "not sure they'd made the right career choice" (p<.001), and 4% of all respondents indicated "definitely not being happy with their career." It is of concern that veterinarians not only have a higher risk of suicide but that they also have fewer support structures. The wide discrepancies between the published risk of suicide for veterinarians and their own views of their risk suggests an inadequate awareness of their own mental health vulnerability which could put them at higher risk.


Solution structure of a minor and transiently formed state of a T4 lysozyme mutant.
Proteins are inherently plastic molecules, whose function often critically depends on excursions between different molecular conformations (conformers). However, a rigorous understanding of the relation between a protein's structure, dynamics and function remains elusive. This is because many of the conformers on its energy landscape are only transiently formed and marginally populated (less than a few per cent of the total number of molecules), so that they cannot be individually characterized by most biophysical tools. Here we study a lysozyme mutant from phage T4 that binds hydrophobic molecules and populates an excited state transiently (about 1 ms) to about 3% at 25 °C (ref. 5). We show that such binding occurs only via the ground state, and present the atomic-level model of the 'invisible', excited state obtained using a combined strategy of relaxation-dispersion NMR (ref. 6) and CS-Rosetta model building that rationalizes this observation. The model was tested using structure-based design calculations identifying point mutants predicted to stabilize the excited state relative to the ground state. In this way a pair of mutations were introduced, inverting the relative populations of the ground and excited states and altering function. Our results suggest a mechanism for the evolution of a protein's function by changing the delicate balance between the states on its energy landscape. More generally, they show that our approach can generate and validate models of excited protein states.


Formation of CaUO2(CO3)32- and Ca2UO2(CO3)3(aq) complexes at variable temperatures (10-70 °C).
The ternary complexation of calcium uranyl tricarbonate species, CaUO2(CO3)32- and Ca2UO2(CO3)3(aq), which are the predominant U(vi) complexes in groundwater and seawater, was investigated at variable temperatures from 10 to 70 °C. Time-resolved laser fluorescence spectroscopy (TRLFS), calcium ion-selective electrode potentiometry, and ultraviolet/visible (UV/Vis) absorption spectroscopy were complementarily employed to determine the formation constants (log Kx13, x = 1 and 2 for mono- and dicalcium complexes, respectively). at infinite dilution (zero ionic strength) was determined by correction using specific ion interaction theory (SIT), and an increasing tendency of with temperature was observed. In addition, the molar enthalpy of complexation (ΔrHm) was measured by calorimetry at 25 °C. Based on thermodynamic data obtained in this work, the approximation models were examined for the prediction of the temperature effect on the complexation, and the constant enthalpy approximation with the chemical complexation reaction modified to an isoelectric reaction showed a satisfactory prediction of in the temperature range of 10-70 °C. Finally, the results of U(vi) speciation in groundwater indicated that the dominance of calcium uranyl tricarbonate complexes would be weakened at elevated temperatures by the strongly enhanced hydrolysis of U(vi).


[The transgenerational mechanisms in developmental programming of metabolic diseases].
Human epidemiological and experimental animal studies have shown that suboptimal environments in the womb and during early neonatal life alter growth and may program offspring susceptibility to lifelong health problems. One of the most interesting and significant feature of developmental programming is the evidence that adverse consequences of altered intrauterine environments can be passed from first generation to second generation offspring. To obtain the transgenerational phenotype, a negative environment is required during fetal or early neonatal life, the physiologic phenotype or disease can be transmitted through the germ line and the subsequent generations are not directly exposed to the environmental factor. The hypothesis has become well accepted by compelling animal studies that define the outcome of specific challenges such as: 1) nutrient restriction or overfeeding during pregnancy and lactation; 2) uterine blood flow restriction; 3) fetal exposure to inappropriately high levels of glucocorticoids, and 4) experimental maternal diabetes. Maternal protein restriction in the rat adversely affects glucose metabolism of male and female second generation offspring in a gender and developmental time window-specific manner. Other studies have proved transgenerational passage of effects resulting from treatment of pregnant rats with dexamethasone by either maternal or paternal lines. First generation female diabetic offspring of F0 rats treated with streptozotocin during pregnancy had F2 offspring with altered glucose and carbohydrate metabolism. The studies suggest that the mechanisms involved in developmental programming are likely epigenetic rather than due to DNA sequence mutations. Many individuals all over the world experience undernutrition, stress, hyperglycemia and other negative environmental factors during pregnancy and/or lactation. Insult during this critical period of development may induce malprogramming and adversely alter not only the F1 generation but also future generations. Preventing or treating these conditions will help to minimize the risk of transmission of metabolic diseases to future generations.


Risk assessment of PBDEs and PAHs in house dust in Kocaeli, Turkey: levels and sources.
Indoor dust samples were collected from 40 homes in Kocaeli, Turkey and were analyzed simultaneously for 14 polybrominated diphenyl ethers (PBDEs) and 16 poly aromatic hydrocarbons (PAHs) isomers. The total concentrations of PBDEs (Σ14PBDEs) ranged from 29.32 to 4790 ng g-1, with a median of 316.1 ng g-1, while the total indoor dust concentrations of 16 PAHs (Σ16PAHs) extending over three to four orders of magnitude ranged from 85.91 to 40,359 ng g-1 with a median value of 2489 ng g-1. Although deca-PBDE products (BDE-209) were the principal source of PBDEs contamination in the homes (median, 138.3 ng g-1), the correlation in the homes was indicative of similar sources for both the commercial penta and deca-PBDE formulas. The PAHs diagnostic ratios indicated that the main sources of PAHs measured in the indoor samples could be coal/biomass combustion, smoking, and cooking emissions. For children and adults, the contributions to ∑14PBDEs exposure were approximately 93 and 25 % for the ingestion of indoor dust, and 7 and 75 % for dermal contact. Exposure to ∑16PAHs through dermal contact was the dominant route for both children (90.6 %) and adults (99.7 %). For both groups, exposure by way of inhalation of indoor dust contaminated with PBDEs and PAHs was negligible. The hazard index (HI) values for BDE-47, BDE-99, BDE-153, and BDE-209 were lower than the safe limit of 1, and this result suggested that none of the population groups would be likely to experience potential health risk due to exposure to PBDEs from indoor dust in the study area. Considering only ingestion + dermal contact, the carcinogenic risk levels of both B2 PAHs and BDE-209 for adults were 6.2 × 10-5 in the US EPA safe limit range while those for children were 5.6 × 10-4 and slightly higher than the US EPA safe limit range (1 × 10-6 and 1 × 10-4). Certain precautions should be considered for children.


Gender and age differences in transthoracic bioimpedance.
Thoracic impedance consists of a constant baseline component Z0 and a time-variable component delta Z which represents the impedance change related to the cardiac cycle. The maximum part of delta Z [(dZ/dt)max] represents the peak of the ascending aortic blood flow. Measurements of basal thoracic impedance are affected by structural and anatomical differences in the thorax related to sex and ageing. This component is a variable in the denominator of Sramek's formula which is used for calculating stroke volume. The aim of this study was to elucidate the question as to whether the age- and sex-related variation in basal impedance may affect bioimpedance measurements of stroke volume. The study comprised 111 healthy subjects (55 males and 56 females) of ages between 20 and 69 years, divided according to age decades into five groups each of males and females. Stroke volume index (SI), Z0 and (dZ/dt)max were measured in every subject, using transthoracic bioimpedance cardiography. Z0 and (dZ/dt)max had significantly higher values in females than in males in every age group except the oldest one in the case of Z0 and the oldest two groups in the case of (dZ/dt)max. Stroke index showed no significant sex difference, although the higher Z0 in females may underestimate the values of stroke index. Elevation of (dZ/dt)max in females may therefore reflect a positive relation to Z0 rather than higher flow rates. Since Z0 and (dZ/dt)max are variants in opposite positions in Sramek's formula (denominator and numerator, respectively), this functional relationship may keep the bioimpedance measurements from being affected by the sex- and age-related changes in Z0.


Complete assignment of the 1H and 13C NMR spectra of antimicrobial 4-arylamino- 3-nitrocoumarin derivatives.
Herein, we describe the synthesis and complete assignment of the (1)H and (13)C NMR chemical shifts of a series of antimicrobial 4-arylamino-3-nitrocoumarin derivatives based on a combination of (1)H and (13)C NMR, (1)H-(1)H-COSY, NOESY, HSQC and HMBC experiments. Conformational effects upon the chemical shifts of the coumarin moiety arising from the anisotropy of the aryl side group are briefly discussed. This study provides the first complete and fully assigned NMR data for this important group of antimicrobial compounds and bridges the gap existing in the literature with regard to NMR structural data for 4-arylamino-3-nitrocoumarins.


Construction of Red Fox Chromosomal Fragments from the Short-Read Genome Assembly.
The genome of a red fox (Vulpes vulpes) was recently sequenced and assembled using next-generation sequencing (NGS). The assembly is of high quality, with 94X coverage and a scaffold N50 of 11.8 Mbp, but is split into 676,878 scaffolds, some of which are likely to contain assembly errors. Fragmentation and misassembly hinder accurate gene prediction and downstream analysis such as the identification of loci under selection. Therefore, assembly of the genome into chromosome-scale fragments was an important step towards developing this genomic model. Scaffolds from the assembly were aligned to the dog reference genome and compared to the alignment of an outgroup genome (cat) against the dog to identify syntenic sequences among species. The program Reference-Assisted Chromosome Assembly (RACA) then integrated the comparative alignment with the mapping of the raw sequencing reads generated during assembly against the fox scaffolds. The 128 sequence fragments RACA assembled were compared to the fox meiotic linkage map to guide the construction of 40 chromosomal fragments. This computational approach to assembly was facilitated by prior research in comparative mammalian genomics, and the continued improvement of the red fox genome can in turn offer insight into canid and carnivore chromosome evolution. This assembly is also necessary for advancing genetic research in foxes and other canids.


At what cost does a potential survival advantage of bevacizumab make sense for the primary treatment of ovarian cancer? A cost-effectiveness analysis.
To determine whether the addition of bevacizumab to paclitaxel and carboplatin for the primary treatment of advanced ovarian cancer can be cost effective. A cost-effectiveness analysis compared the three arms of the Gynecologic Oncology Group (GOG) 218 study (paclitaxel plus carboplatin [PC], PC plus bevacizumab [PCB], and PCB plus bevacizumab maintenance [PCB+B]). Actual and estimated costs of treatment plus the potential costs of complications were established for each strategy. Progression-free survival (PFS) and bowel perforation rates were taken from recently reported results of GOG 218. Sensitivity analysis was performed for pertinent uncertainties in the model. Incremental cost-effectiveness ratios (ICERs) per progression-free life-year saved (PF-LYS) were estimated. For the 600 patients entered onto each arm of GOG 218 at the baseline estimates of PFS and bowel perforation, the cost of PC was $2.5 million, compared with $21.4 million for PCB and $78.3 million for PCB+B. These costs led to an ICER of $479,712 per PF-LYS for PCB and $401,088 per PF-LYS for PCB+B. When the cost of bevacizumab was reduced to 25% of baseline, the ICER of PCB+B fell below $100,000 per PF-LYS. ICERs were not substantially reduced when the perforation rate was equal across all arms. The addition of bevacizumab to standard chemotherapy in patients with advanced ovarian cancer is not cost effective. Treatment with maintenance bevacizumab leads to improved PFS but is associated with both direct and indirect costs. The cost effectiveness of bevacizumab in the adjuvant treatment of ovarian cancer is primarily dependent on drug costs.


Efficacy, immunogenicity and safety of heptavalent pneumococcal conjugate vaccine in low birth weight and preterm infants.
To determine the efficacy, immunogenicity and safety of the heptavalent CRM197 pneumococcal conjugate vaccine (PCV) in low birth weight (LBW) and preterm (PT) infants against invasive pneumococcal disease caused by vaccine types. In a randomized double blind trial of 37,868 infants given either PCV or meningococcal type C conjugate vaccine (MCV), 1756 infants <750 g <2500 g (LBW) and 4340 infants from 32 to <38 weeks old (PT) were identified. Risk of invasive pneumococcal disease in LBW and PT infants was compared with risk in normal birth weight (NBW) and full term (FT) infants. Local and systemic events observed within 48 h of recent vaccine were assessed by telephone interviews and similar comparisons made. Premature infant Emergency Department visits and hospitalization were also identified and compared with FT and NBW infants. Initiation of immunization and intervals between doses were similar for all groups. The risk ratio for invasive pneumococcal diseases for LBW infants compared with NBW infants was 2.6 (P = 0.03), and for PT compared with FT infants the risk ratio was 1.6 (P = 0.06). Vaccine efficacy for both groups was 100%. PCV was as immunogenic in LBW and PT as in NBW and FT infants. Fever and local events after PCV vaccination were similar when adjusted for clustering among multiple doses per child. When stratified for individual doses there was more redness and swelling for LBW infants and more swelling for PT infants after Dose 3. Isolated local and systemic reactions were more commonly seen with PCV than with MCV, a pattern similar to that in NBW and FT infants. Hospitalization rates were similar for PCV and MCV recipients. These data support the use of PCV in LBW infants and PT infants.


The use of transient chromatin immunoprecipitation assays to test models for E2F1-specific transcriptional activation.
The E2F family of transcription factors regulates the expression of genes involved in cell cycle progression, DNA synthesis, repair, and recombination, and a variety of other cellular processes. Although E2F proteins are often redundant in function, specificity of binding and activity can occur. For example, E2F1, but not other E2F family members, was shown previously to bind the murine carboxylesterase promoter in chromatin immunoprecipitation studies. This promoter region lacks a consensus E2F binding site, suggesting that E2F1 may be recruited to the DNA in a unique fashion. To further investigate this E2F1-specific binding, we have employed a "transient chromatin immunoprecipitation" approach. Using various deletions and mutations of the promoter region, we localized the E2F1-specific binding site and demonstrated that it was required for E2F1-mediated transcription of the carboxylesterase promoter. The identified site was similar to the 8-bp consensus E2F site but differed from the consensus at a crucial position. To address whether E2F1 directly bound to this non-consensus site, we demonstrated that the DNA binding domain of E2F1 is necessary for E2F1-mediated activation of the carboxylesterase promoter. Interestingly, a "UP" mutation of the site, making it more similar to the consensus element, did not improve the ability of E2F1 to bind the promoter. Rather, E2F1 could no longer bind to the carboxylesterase promoter that contained the consensus E2F site. We propose a model in which E2F1-specific regulation of the carboxylesterase promoter requires both E2F1/DNA interactions and protein-protein interaction between E2F1 and a factor that binds adjacent to the non-consensus site.


Perfusion of surgical cavity wall enhancement in early post-treatment MR imaging may stratify the time-to-progression in glioblastoma.
To determine if perfusion in surgical cavity wall enhancement (SCWE) obtained in early post-treatment MR imaging can stratify time-to-progression (TTP) in glioblastoma. This study enrolled 60 glioblastoma patients with more than 5-mm-thick SCWEs as detected on contrast-enhanced MR imaging after concurrent chemoradiation therapy. Two independent readers categorized the shape and perfusion state of SCWEs as nodular or non-nodular and as having positive or negative perfusion compared with the contralateral grey matter on arterial spin labeling (ASL). The perfusion fraction on ASL within the contrast-enhancing lesion was calculated. The independent predictability of TTP was analyzed using the Kaplan-Meier method and Cox proportional hazards modelling. The perfusion fraction was higher in the non-progression group, significantly for reader 2 (P = 0.03) and borderline significantly for reader 1 (P = 0.08). A positive perfusion state and (P = 0.02) a higher perfusion fraction of the SCWE were found to become an independent predictor of longer TTP (P = 0.001 for reader 1 and P < 0.001 for reader 2). The contrast enhancement pattern did not become a TTP predictor. Assessment of perfusion in early post-treatment MR imaging can stratify TTP in patients with glioblastoma for adjuvant temozolomide therapy. Positive perfusion in SCWEs can become a predictor of a longer TTP.


Isoliquiritigenin Ameliorates Acute Pancreatitis in Mice via Inhibition of Oxidative Stress and Modulation of the Nrf2/HO-1 Pathway.
Oxidative stress plays a crucial role in the pathogenesis of acute pancreatitis (AP). Isoliquiritigenin (ISL) is a flavonoid monomer with confirmed antioxidant activity. However, the specific effects of ISL on AP have not been determined. In this study, we aimed to investigate the protective effect of ISL on AP using two mouse models. In the caerulein-induced mild acute pancreatitis (MAP) model, dynamic changes in oxidative stress injury of the pancreatic tissue were observed after AP onset. We found that ISL administration reduced serum amylase and lipase levels and alleviated the histopathological manifestations of pancreatic tissue in a dose-dependent manner. Meanwhile, ISL decreased the oxidative stress injury and increased the protein expression of the Nrf2/HO-1 pathway. In addition, after administering a Nrf2 inhibitor (ML385) or HO-1 inhibitor (zinc protoporphyrin) to block the Nrf2/HO-1 pathway, we failed to observe the protective effects of ISL on AP in mice. Furthermore, we found that ISL mitigated the severity of pancreatic tissue injury and pancreatitis-associated lung injury in a severe acute pancreatitis model induced by L-arginine. Taken together, our data for the first time confirmed the protective effects of ISL on AP in mice via inhibition of oxidative stress and modulation of the Nrf2/HO-1 pathway.


Evidence of balanced diversity at the chicken interleukin 4 receptor alpha chain locus.
The comparative analysis of genome sequences emerging for several avian species with the fully sequenced chicken genome enables the genome-wide investigation of selective processes in functionally important chicken genes. In particular, because of pathogenic challenges it is expected that genes involved in the chicken immune system are subject to particularly strong adaptive pressure. Signatures of selection detected by inter-species comparison may then be investigated at the population level in global chicken populations to highlight potentially relevant functional polymorphisms. Comparative evolutionary analysis of chicken (Gallus gallus) and zebra finch (Taeniopygia guttata) genes identified interleukin 4 receptor alpha-chain (IL-4Ralpha), a key cytokine receptor as a candidate with a significant excess of substitutions at nonsynonymous sites, suggestive of adaptive evolution. Resequencing and detailed population genetic analysis of this gene in diverse village chickens from Asia and Africa, commercial broilers, and in outgroup species red jungle fowl (JF), grey JF, Ceylon JF, green JF, grey francolin and bamboo partridge, suggested elevated and balanced diversity across all populations at this gene, acting to preserve different high-frequency alleles at two nonsynonymous sites. Haplotype networks indicate that red JF is the primary contributor of diversity at chicken IL-4Ralpha: the signature of variation observed here may be due to the effects of domestication, admixture and introgression, which produce high diversity. However, this gene is a key cytokine-binding receptor in the immune system, so balancing selection related to the host response to pathogens cannot be excluded.


'Miracle baby': an outcome of multidisciplinary approach to neurotrauma in pregnancy.
Traumatic brain injury (TBI) warranting neurosurgical intervention in the pregnant population is a rarity. We describe a case of a 27-year-old woman who at 13 weeks of gestation presented with multiple traumas having been involved in a near fatal road traffic accident. Glasgow Coma Scale was 6/15. CT brain showed extensive haemorrhagic contusions, diffuse brain swelling and multiple skull and facial fractures. Decompressive craniectomy was performed to control her intracranial pressure during her management in the intensive care. A viable intrauterine pregnancy was confirmed and progressed as maternal stabilisation and rehabilitation continued. At 35+3 weeks a 2770 g male child was delivered via emergency caesarean section after spontaneous onset of labour. The child had no detectable abnormalities and is clinically well. Eight months post-TBI the patient continues to make gradual improvements but is left with severe cognitive impairment and currently undergoing rehabilitation. A multidisciplinary approach was adopted in the management of this patient.


Regional timing of myocardial shortening is related to prestretch from atrial contraction: assessment by high temporal resolution MRI tagging in humans.
Earlier studies have shown substantial nonuniformity in normal left ventricular (LV) myocardial function concerning both the degree of shortening and timing of shortening. We hypothesized that nonuniform LV function may be related to nonuniform prestretch induced by atrial contraction. Eleven healthy human subjects were studied using MRI myocardial tagging and strain analysis. The amount of circumferential prestretch was assessed in 30 LV segments. Prestretch was defined as the difference in strain between end diastole (at ECG R wave) and diastasis. Furthermore, both the degree of shortening (quantified as peak circumferential shortening, peak systolic shortening rate, and amount of postsystolic shortening) and timing of shortening (quantified as the onset time of shortening and time to peak shortening) were assessed. LV prestretch was found to be nonuniform, with the highest values in the lateral wall. The amount of segmental prestretch correlated significantly with peak shortening (r = 0.79), peak shortening rate (r = 0.50), amount of postsystolic shortening (r = 0.67), onset time of shortening (r = -0.57), and time to peak shortening (r = 0.71) (P < 0.001 for each of these relations). These relations may be explained by regional differences in wall stress or by a regional Frank-Starling effect. The correlation between timing of shortening and prestretch demonstrates that mechanical timing is not determined by electrical phenomena alone. In conclusion, regional variation in LV function correlates with the nonuniform prestretch from atrial contraction.