Staging of Prostate Cancer: Role of Multiparametric Magnetic Resonance Imaging in Different Risk Classes.
Using multiparametric magnetic resonance imaging, it is now possible to diagnose prostate cancer and categorize its risk. As it can accurately determine the extracapsu- lar extension of the tumor, invasion of seminal vesicles, involvement of lymph nodes, and the potential presence of bone metastases, multiparametric magnetic resonance imaging plays a crucial role not only in the diagnosis but also in the local staging of prostate cancer. The patients with a history of negative biopsy/increasing prostate- specific antigen and the existence of further data supporting its use in biopsy-naive patients and active surveillance are the most blatant indications for multiparametric magnetic resonance imaging in guidelines. The traditional clinical examination, pros- tate-specific antigen tests, and systematic biopsy are all enhanced by multiparametric magnetic resonance imaging, which will miss certain cancers due to insufficient size or changes in tissue density. The use of multiparametric magnetic resonance imaging is expected to rise, and further advances in the method will be crucial for the secure adoption of targeted therapeutic ideas. Here, we give a succinct overview of multipa- rametric magnetic resonance imaging's application to the identification and risk clas- sification of prostate cancer.


Activating mutation of adenylyl cyclase reverses its inhibition by G proteins.
We have implemented a yeast genetic selection developed previously by our laboratory to identify mutant mammalian type V adenylyl cyclases insensitive to inhibition by G(ialpha.) One mutation isolated was localized to the first cytoplasmic domain at a Phe residue (position 400), which is conserved in all nine isoforms of membrane-bound mammalian adenylyl cyclase. Biochemical characterization of the F400Y mutant revealed a dramatic conversion of the G(ialpha) response from inhibitory to stimulatory. This mutation results in additional activating effects. The mutant exhibits an enhanced sensitivity toward activation by either G(salpha) or forskolin. Synergism between G(salpha) and forskolin is not observed for the F400Y mutant, presumably because the mutant already is in the sensitized state. Additionally, an enhancement of the basal unstimulated activity was observed. This mutation, which is the first demonstration of an activating point in a mammalian adenylyl cyclase, mimics a sensitized conformation of the wild-type enzyme that underlies the synergism between stimulatory inputs, and additionally, removes the inhibitory regulatory input provided by G(ialpha). Because sensitizing adenylyl cyclase toward its stimulators can have profound biological implications, this raises the possibility that naturally occurring mutations resembling those at the Phe400 residue may be associated with human disease states.


Seropositivity in Dutch Crohn's disease patients against primed nude mouse lymph nodes, and the difference with lymphocytotoxic antibodies.
Sera from patients with Crohn's disease have been reported to show positive immunofluorescence with lymph nodes of nude mice primed with a filtrate of intestinal tissue affected with Crohn's disease. An indirect immunofluorescence assay was used to test sera of 63 unrelated patients with Crohn's disease, 21 with ulcerative colitis and 36 control subjects against lymph nodes of athymic nude (nu/nu) mice which had been injected with Crohn's disease and ulcerative colitis intestinal tissue filtrates. Forty nine per cent of Crohn's disease patients, 10% of ulcerative colitis patients and 3% of control sera reacted against lymph nodes of mice injected injected with ulcerative colitis intestinal tissue filtrates, 18% of Crohn's disease sera were with intestinal tissue homogenate from Dutch Crohn's patients. With the lymph nodes of mice injected with ulcerative colitis intestinal tissue filtrates, 18% of Crohn's disease sera were positive, whereas all ulcerative colitis and control sera were negative. Lymph nodes from 18 of the 19 mice injected with Crohn's disease tissue filtrates reacted with Crohn's disease sera, whereas only three of these 19 mice reacted with ulcerative colitis sera. A comparative study, carried out in parallel with Crohn's disease filtrate induced hyperplastic lymph nodes from the Bilthoven colony (W2) and from the New York colony (E671) using sera from 54 Crohn's disease patients from Leiden, showed immunoreactivity with 44 and 57% of the Crohn's disease sera against the two hyperplastic lymph nodes. Thirty six of the 54 Crohn's disease sera (67%) reacted with either or both lymph nodes. Only 11% of the Crohn's disease sera which were examined for immunofluorescence and lymphocytotoxic antibodies had lymphocytotoxic antibodies, whereas 40% and 46% of the same sera showed positive immunofluorescence against E671 and W2, respectively. Absorption studies indicated that lymphocytotoxic antibodies activity and the immunofluorescence against the primed nude mouse lymph node are mediated by different serum antibodies in Crohn's disease. The reproducibility of the nude mouse immunofluorescence test system for a preferential immunoreactivity of Crohn's disease sera against Crohn's disease tissue primed murine lymph nodes has been confirmed by the present study. Further studies are necessary to find out whether crossreactive antigen(s) as recognised by some of the Crohn's disease sera in mice injected with ulcerative colitis tissue filtrate is similar to the antigen(s) detected by Crohn's disease sera in mice injected with Crohn's disease tissue filtrates.


Effects of fiber alignment on stem cells-fibrous scaffold interactions.
Considerable research has been focused on the utilization of electrospun ultrafine fibers to construct biomimetic scaffolds for tissue engineering applications. However, there is still a limited understanding of the effects of fiber alignment on stem cells-fibrous scaffold interactions and their role in deciding the fate of stem cells and scaffolds. Here, we tracked the real-time interactions between mesenchymal stem cells (MSCs) and aligned/non-aligned electrospun PCL-gelatin ultrafine fibers using time-lapse microscopy, and investigated the effects of fiber alignment on the behaviour of cell spreading, migration and differentiation, and scaffold remodeling. Cells were found to spread and migrate with increased velocity in the direction of aligned fibers as compared to non-aligned ones. Moreover, enhanced interactions between cells and aligned fibers were found to facilitate the infiltration of cells into the interior of fibrous constructs, which further led to enhanced micro-integration and scaffold remodeling as well as increased gene expression of late osteogenic markers as compared to non-aligned fibrous constructs. This study advances our understanding of the effects of fiber alignment on stem cells-fibrous scaffold interactions, and will facilitate the design of fibrous scaffolds with enhanced cell-matrix interactions for tissue engineering applications.


Merkel cell carcinoma with an unusual immunohistochemical profile.
The clinical and morphological picture of Merkel cell carcinoma (MCC) may be rather challenging; therefore, the immunohistochemical profile plays a relevant role in confirming the microscopic diagnosis. A panel of antibodies including cytokeratins 20, 7 and epithelial membrane antigen, and neuron-specific enolase is used in confirming the morphological diagnosis of MCC. The majority of MCCs express CK20 and are CK7-negative. Herein, we present a case of primary cutaneous neuroendocrine carcinoma with an atypical immunohistochemical pattern. A 83-years old woman presented with a painless plaque, red to violaceous in colour, located in the leg. The skin tumor was excided, formalin-fixed and paraffinembedded. Tissue sections were immunostained with a panel of antibodies routinely utilized in complex primary skin tumors for evidencing epithelial and neuroendocrine differentiation of tumor cells. The neuroendocrine differentiation of tumor cells was evidenced by their immunoreactivity for synaptophysin, chromograninA and neuron-specific enolase. Tumor cells also showed diffuse cytoplasmic staining for CK7. No immunoreactivity was detected for CK20 and thyroid transcription factor-1. Our data, together with previous rare reports of CK20-/CK7+ MCCs, lay stress on the importance of routinely utilizing a panel of antibodies in the differential diagnosis of complex primary carcinomas of the skin and may have important implications in expanding the differential diagnosis of skin tumors. In particular, caution should be taken in excluding the diagnosis of MCC only on the basis of the absence of reactivity of tumor cells for CK20, favouring the wrong diagnosis of less aggressive skin tumors.


A simple procedure for isolation of eukaryotic mRNA polyadenylation factors.
We have devised a simple chromatographic procedure which isolates five polyadenylation factors that are required for polyadenylation of eukaryotic mRNA. The factors were separated from each other by fractionation of HeLa cell nuclear extract in two consecutive chromatographic steps. RNA cleavage at the L3 polyadenylation site of human adenovirus 2 required at least four factors. Addition of adenosine residues required only two of these factors. The fractionation procedure separates two components that are both likely to be poly(A) polymerases. The candidate poly(A) polymerases were interchangeable and participated during both RNA cleavage and adenosine addition. They were discriminated from each other by chromatographic properties, heat sensitivity and divalent cation requirement. We have compared our data with published information and have been able to correlate the activities that we have isolated to previously identified polyadenylation factors. However, we have not been able to assign one of the candidate poly(A) polymerases to a previously identified poly(A) polymerase. This simple fractionation procedure can be used for generating an in vitro reconstituted system for polyadenylation within a short period of time.


Managing the middle: A shift in conservation priorities based on the global human modification gradient.
An increasing number of international initiatives aim to reconcile development with conservation. Crucial to successful implementation of these initiatives is a comprehensive understanding of the current ecological condition of landscapes and their spatial distributions. Here, we provide a cumulative measure of human modification of terrestrial lands based on modeling the physical extents of 13 anthropogenic stressors and their estimated impacts using spatially explicit global datasets with a median year of 2016. We quantified the degree of land modification and the amount and spatial configuration of low modified lands (i.e., natural areas relatively free from human alteration) across all ecoregions and biomes. We identified that fewer unmodified lands remain than previously reported and that most of the world is in a state of intermediate modification, with 52% of ecoregions classified as moderately modified. Given that these moderately modified ecoregions fall within critical land use thresholds, we propose that they warrant elevated attention and require proactive spatial planning to maintain biodiversity and ecosystem function before important environmental values are lost.


The impact of different concentrations of hyaluronic acid on the pasting and microstructural properties of corn starch.
Starch aging in starchy foods is a major problem affecting their quality. In order to improve the viscosity and textural properties of native starch gelatinization and retrogradation, this study investigated the effect of hyaluronic acid (HA) at different concentrations (2 %, 4 %, 6 % w/w) on the pasting and microstructure of corn starch (CS). The findings revealed that the addition of HA significantly enhanced the peak viscosity, solubility, and water-holding capacity of the CS-HA mixtures. Moreover, it reduced the pasting temperature, swelling force, and leaching of amylose. All the mixtures exhibited shear thinning and thixotropic properties. The CS-HA mixtures created a thicker pseudoplastic system with significantly enhanced shear stability. The structures of the mixtures were characterized using Fourier transform infrared spectroscopy and scanning electron microscopy. It was observed that HA effectively inhibited short-term retrogradation of starch, enhanced the interaction between CS and HA, and formed a dense honeycomb three-dimensional mesh structure. In conclusion, as a novel anionic hydrocolloid, HA holds great potential to improve the retrogradation properties of starch-based products.


Legal requirements for the quality of herbal substances and herbal preparations for the manufacturing of herbal medicinal products in the European union.
In the European Union (EU) herbal medicinal products have become increasingly important. This is, for instance, underlined by the recent introduction of a simplified procedure in the Member States of the EU allowing the registration of herbal medicinal products which fulfill the criteria of a traditional herbal medicinal product, i.e., sufficient evidence of its medicinal use throughout a period of at least 30 years for products in the EU and at least 15 years within the EU and 15 years elsewhere for products outside the EU. With regard to the manufacturing of these products and their quality, applications of traditional herbal medicinal products have to fulfil the same requirements as applications for a marketing authorization. The quality of herbal substances as well as herbal preparations will be determined by the availability of modern science-based public monographs in the European Pharmacopoeia and their equivalents developed by the pharmaceutical industry. The standards put forward in these monographs must allow us not only to define the quality of these products, but also to eliminate dangerous counterfeit, substandard, adulterated and contaminated (traditional) herbal medicinal products. The usefulness of these monographs to implement the criteria on quality and specifications put forward for these products in the different guidelines of the European Medicines Agency (EMEA) is discussed.


Network Enrichment Significance Testing in Brain-Phenotype Association Studies.
Functional networks often guide our interpretation of spatial maps of brain-phenotype associations. However, methods for assessing enrichment of associations within networks of interest have varied in terms of both scientific rigor and underlying assumptions. While some approaches have relied on subjective interpretations, others have made unrealistic assumptions about the spatial structure of imaging data, leading to inflated false positive rates. We seek to address this gap in existing methodology by borrowing insight from a method widely used in genomics research for testing enrichment of associations between a set of genes and a phenotype of interest. We propose Network Enrichment Significance Testing (NEST), a flexible framework for testing the specificity of brain-phenotype associations to functional networks or other sub-regions of the brain. We apply NEST to study phenotype associations with structural and functional brain imaging data from a large-scale neurodevelopmental cohort study.


Sexual dimorphism in anxiety is programmed in-utero by sex-steroids: Proof of concept using a disease-model and stress responses to COVID pandemic.
Sex differences in the psychological impact of the COVID-19 pandemic have been consistently reported, showing disproportionally high rates of anxiety/distress in women relative to men. The mechanisms underlying this sexual dimorphism remain unclear. The present study queries the potential protective role of early hyperandrogenism on brain development. A natural model of sex-steroids abnormality, classic congenital adrenal hyperplasia (CAH), was used to test this question. CAH is characterized by adrenal androgen overproduction in utero (prenatal) through the neonatal critical developmental period. An online survey collected information on subjective responses to the COVID-19 pandemic. Matched on demographic variables, 60 adults carrying a diagnosis of classic CAH and 240 adults from the general population (non-CAH) were compared on levels of anxiety/distress in the first year of the COVID-19 pandemic (May 2020-April 2021). Structural Equation Modeling was used to test the modulation by CAH status of Sex effects on anxiety/distress. Results revealed lower levels of anxiety/distress in the female CAH group compared to the other 3 groups (male CAH, female non-CAH, and male non-CAH). This finding suggests that pre-neonatal hyperandrogenism might impact the development of neural circuits underlying anxiety processes, in a way that enhances resilience to chronic stress.


[Duration of induced seizures during selective pharyngeal brain cooling].
Whole body hypothermia can be used to treat the injured brain (e.g. after hypoxic events). Side effects include hemodynamic instability, coagulopathy and infection. Because of these side effects it appears reasonable to cool the brain selectively (selective brain cooling, SBC) without changing the core temperature. A new animal model was used to demonstrate SBC from the pharynx and to examine effects of SBC on the duration of pharmacologically induced seizure activity. Sprague-Dawley rats (n=18, 12 successful experiments) were sedated and mechanically ventilated. Invasive blood pressure monitoring was instituted and blood gases were drawn to evaluate the arterial blood gas status. Electrical brain activity was recorded using a microneedle in the extracellular compartment of the rat brain cortex. Cooled water was circulated through a small tubing into and out of the pharynx of the animals. The cortical as well as the rectal temperature were recorded. After the injection of a single dose of bicuculline (1 mg/kg i.v.) per animal the duration of the induced seizure activity was measured and compared with the temperature prior to the induction of seizure activity. The cortical blood flow (CBF) was detected using intra tissue Doppler signals in the rat cortex in the same location as the EP-study. The influence of a brain temperature reduction between 36.5 degrees to 31.5 degrees C on the seizure duration was examined. There was a positive correlation between the seizure duration and the cortical temperature (r=0.64). Also the CBF was increased during seizure activity (p=0.02) and the increase correlated weakly with cortical temperature (r=0.18). The core temperature remained in the normothermic range (36.9+/-0.7 degrees C) Conclusion: The duration of induced seizures correlates with local brain temperature. In the future further studies should examine the efficiency of induced (selective) brain cooling to treat prolonged seizure activity.


Racially and ethnically diverse schools and adolescent romantic relationships.
Focusing on romantic relationships, which are often seen as a barometer of social distance, this analysis investigates how adolescents from different racial-ethnic and gender groups respond when they attend diverse schools with many opportunities for inter-racial-ethnic dating. Which groups respond by forming inter-racial-ethnic relationships, and which groups appear to "work around" opportunities for inter-racial-ethnic dating by forming more same-race-ethnicity relationships outside of school boundaries? Most prior studies have analyzed only relationships within schools and, therefore, cannot capture a potentially important way that adolescents express preferences for same-race-ethnicity relationships or work around constraints from other groups' preferences. Using the National Longitudinal Study of Adolescent Health, I find that, when adolescents are in schools with many opportunities for inter-racial-ethnic dating, black females and white males are most likely to form same-race-ethnicity relationships outside of the school; whereas Hispanic males and females are most likely to date across racial-ethnic boundaries within the school.


Circadian genes are expressed during early development in Xenopus laevis.
Circadian oscillators are endogenous time-keeping mechanisms that drive twenty four hour rhythmic changes in gene expression, metabolism, hormone levels, and physical activity. We have examined the developmental expression of genes known to regulate circadian rhythms in order to better understand the ontogeny of the circadian clock in a vertebrate. In this study, genes known to function together in part of the core circadian oscillator mechanism (xPeriod1, xPeriod2, and xBmal1) as well as a rhythmic, clock-controlled gene (xNocturnin) were analyzed using in situ hybridization in embryos from neurula to late tailbud stages. Each transcript was present in the developing nervous system in the brain, eye, olfactory pit, otic vesicle and at lower levels in the spinal cord. These genes were also expressed in the developing somites and heart, but at different developmental times in peripheral tissues (pronephros, cement gland, and posterior mesoderm). No difference was observed in transcript levels or localization when similarly staged embryos maintained in cyclic light were compared at two times of day (dawn and dusk) by in situ hybridization. Quantitation of xBmal1 expression in embryonic eyes was also performed using qRT-PCR. Eyes were isolated at dawn, midday, dusk, and midnight (cylic light). No difference in expression level between time-points was found in stage 31 eyes (p = 0.176) but stage 40 eyes showed significantly increased levels of xBmal1 expression at midnight (RQ = 1.98+/-0.094) when compared to dawn (RQ = 1+/-0.133; p = 0.0004). We hypothesize that when circadian genes are not co-expressed in the same tissue during development that it may indicate pleiotropic functions of these genes that are separate from the timing of circadian rhythm. Our results show that all circadian genes analyzed thus far are present during early brain and eye development, but rhythmic gene expression in the eye is not observed until after stage 31 of development.


An appropriate plot area for analyzing canopy cover and tree species richness in Zagros forests.
In order to make the sampling procedure more efficient and more accurate to study the tree species richness and canopy cover, the appropriate plot size was calculated in the this study. The sampling was carried out using 48 four-hectare plots, each with 13 sub-plots of different plot sizes and 7 one-hectare plots, each with 7 sub-plots. The result of this study showed that 300 ARE plot size was determined as the best area for 1-5% density class, 125 ARE plots for 5-10% class, 150 ARE for 10-25% class, 100 ARE for 25-50% class and 75 ARE plot size to sample >50% density class, in 95% confidence level. Consequently, using 100 ARE sampling plots is suggested for all density classes in central Zagros forests.


Chemical Analysis of the Interface between Hybrid Organic-Inorganic Perovskite and Atomic Layer Deposited Al2O3.
Ultrathin metal oxides prepared by atomic layer deposition (ALD) have gained utmost attention as moisture and thermal stress barrier layers in perovskite solar cells (PSCs). We have recently shown that 10 cycles of ALD Al2O3 deposited directly on top of the CH3NH3PbI3- xCl x perovskite material, are effective in delivering a superior PSC performance with 18% efficiency (compared to 15% of the Al2O3-free cell) with a long-term humidity-stability of more than 60 days. Motivated by these results, the present contribution focuses on the chemical modification which the CH3NH3PbI3- xCl x perovskite undergoes upon growth of ALD Al2O3. Specifically, we combine in situ Infrared (IR) spectroscopy studies during film growth, together with X-ray photoelectron spectroscopy (XPS) analysis of the ALD Al2O3/perovskite interface. The IR-active signature of the NH3+ stretching mode of the perovskite undergoes minimal changes upon exposure to ALD cycles, suggesting no diffusion of ALD precursor and co-reactant (Al(CH3)3 and H2O) into the bulk of the perovskite. However, by analyzing the difference between the IR spectra associated with the Al2O3 coated perovskite and the pristine perovskite, respectively, changes occurring at the surface of perovskite are monitored. The abstraction of either NH3 or CH3NH2 from the perovskite surface is observed as deduced by the development of negative N-H bands associated with its stretching and bending modes. The IR investigations are corroborated by XPS study, confirming the abstraction of CH3NH2 from the perovskite surface, whereas no oxidation of its inorganic framework is observed within the ALD window process investigated in this work. In parallel, the growth of ALD Al2O3 on perovskite is witnessed by the appearance of characteristic IR-active Al-O-Al phonon and (OH)-Al═O stretching modes. Based on the IR and XPS investigations, a plausible growth mechanism of ALD Al2O3 on top of perovskite is presented.


The relationship between obesity and exposure to light at night: cross-sectional analyses of over 100,000 women in the Breakthrough Generations Study.
There has been a worldwide epidemic of obesity in recent decades. In animal studies, there is convincing evidence that light exposure causes weight gain, even when calorie intake and physical activity are held constant. Disruption of sleep and circadian rhythms by exposure to light at night (LAN) might be one mechanism contributing to the rise in obesity, but it has not been well-investigated in humans. Using multinomial logistic regression, we examined the association between exposure to LAN and obesity in questionnaire data from over 100,000 women in the Breakthrough Generations Study, a cohort study of women aged 16 years or older who were living in the United Kingdom and recruited during 2003-2012. The odds of obesity, measured using body mass index, waist:hip ratio, waist:height ratio, and waist circumference, increased with increasing levels of LAN exposure (P < 0.001), even after adjustment for potential confounders such as sleep duration, alcohol intake, physical activity, and current smoking. We found a significant association between LAN exposure and obesity which was not explained by potential confounders we could measure. While the possibility of residual confounding cannot be excluded, the pattern is intriguing, accords with the results of animal experiments, and warrants further investigation.


Global spectral and location effects in auditory perceptual grouping.
An important problem in cognitive and systems neuroscience concerns the extent to which perceptual organization can be explained by "local," peripheral physiological mechanisms, or rather by more "global," central, and higher-level processes. Though central in vision research, this issue has received little attention in the field of audition. One claim is that auditory-perceptual grouping mechanisms, possibly related to visual figure-from-ground segregation or "pop-out," are low level, resulting from local processing in the frequency domain. However, no experiments have been performed specifically to test this question. We examined the effects of perceptual grouping on detection for reversal of two repeated target tones, one constant in frequency (1030 Hz), the other free to vary between trials (1045-8580 Hz). Detection was examined in the presence of a 1000-Hz background tone that repeated between target presentations. By varying the frequency of the high-target tone, this task was designed to modulate grouping between the background and low-target tones, thereby affecting reversal detection. We predicted that at large target frequency differences (deltaf), the high-target tone would segregate from the background and low-target tones, and so render the background and low-target tones less distinct. We found that reversal detection declined from optimal levels with increasing deltaf, and that performance was improved by spatially separating the location of the target and background sounds by at least 32 degrees. These results demonstrate that global frequency integration over at least three octaves occurs through grouping, and that grouping is affected by source location. This implies that auditory-perceptual grouping involves global neural processing, i.e., the participation of neurons with very broad frequency input that are also sensitive to spatial location.


Irradiation of Escherichia coli in the visible spectrum with a tunable organic-dye laser energy source.
Pulsed laser energy was shown to be effective in inhibiting the growth of Escherichia coli. The irradiation source was derived from a tunable organic-dye laser utilizing rhodamine 6G (590 plus or minus 5 nm) solutions as lasing media. The organisms, suspended in nutrient broth, were irradiated both with and without an exogenous photosensitizer. One photosensitizer (toluidine blue) did not appreciably alter the inhibitory effect observed. In the presence of acridine orange, however, some additional growth occurred.


Effect of four chitinase genes on the female fecundity in Sogatella furcifera (Horváth).
The White-Backed Planthopper (WPH), Sogatella furcifera (Horváth), is a destructive rice pest with strong reproductive capacity. To gain insights into the roles of chitinases in the reproductive process of this insect species, this study represents the first-ever endeavor to conduct an in-depth exploration into the reproductive functions of four chitinase genes. In this study, it was observed that four chitinase genes were expressed in female adults, with a relatively high expression level in the ovaries. SfCht2 and SfIDGF1 were highly expressed during later ovarian development. while SfENGase increased and then decreased with ovarian development. SfCht2, SfCht6-2 and SfENGase were highly expressed in fat body on the first and second days after eclosion, whereas SfIDGF1 highest on day 7. Compared with control group, Silencing four chitinase genes inhibited ovarian development and significantly shortened the oviposition period of S. furcifera, reducing egg-laying capacity but not affecting egg hatching. The detection demonstrated that the expression levels of SfVg, SfVgR and 70%-90% JH signaling pathway-related reproductive genes was significantly down-regulated. Moreover, SfCht6-2 and SfENGase significantly affected the expression levels of TOR signaling pathway genes. SfENGase had the ability to impact nutrient signaling pathways and fatty acid metabolism, repressing vitellogenin synthesis and ultimately influencing ovarian development of S. furcifera. Overall, this study provides insight into the function of chitinases in insect fecundity and is of great significance for enriching the cognition of insect chitinase function. They will become the suitable target genes for controling controlling the most destructive rice planthoppers. This article is protected by copyright. All rights reserved.


Early Ongoing Speciation of Ogataea uvarum Sp. Nov. Within the Grape Ecosystem Revealed by the Internal Variability Among the rDNA Operon Repeats.
A yeast strain was isolated during a study on vineyard-associated yeast strains from Apulia in Southern Italy. ITS and LSU D1/D2 rDNA sequences showed this strain not to belong to any known species and was described as the type strain of Ogataea uvarum sp. nov., a close relative of O. philodendri. Several secondary peaks appeared in the sequences, suggesting internal heterogeneity among the copies of the rDNA. This hypothesis was tested by sequencing single clones of the marker region. The analyses showed different levels of variability throughout the operon with differences between the rRNA encoding genes and the internally transcribed regions. O. uvarum and O. philodendri share high frequency variants, i.e., variants frequently found in many clones, whereas there is a large variability of the low frequency polymorphisms, suggesting that the mechanism of homogenization is more active with the former than with the latter type of variation. These findings indicate that low frequency variants are detected in Sanger sequencing as secondary peaks whereas in Next Generation Sequencing (NGS) of metagenomics DNA would lead to an overestimate of the alpha diversity. For the first time in our knowledge, this investigation shed light on the variation of the copy number of the rDNA cistron during the yeast speciation process. These polymorphisms can be used to investigate on the processes occurring in these taxonomic markers during the separation of fungal species, it being a genetic process highly frequent in the complex microbial ecosystem existing in grape, must and wine.


Arachidonic Acid Kills Staphylococcus aureus through a Lipid Peroxidation Mechanism.
Staphylococcus aureus infects every niche of the human host. In response to microbial infection, vertebrates have an arsenal of antimicrobial compounds that inhibit bacterial growth or kill bacterial cells. One class of antimicrobial compounds consists of polyunsaturated fatty acids, which are highly abundant in eukaryotes and encountered by S. aureus at the host-pathogen interface. Arachidonic acid (AA) is one of the most abundant polyunsaturated fatty acids in vertebrates and is released in large amounts during the oxidative burst. Most of the released AA is converted to bioactive signaling molecules, but, independently of its role in inflammatory signaling, AA is toxic to S. aureus Here, we report that AA kills S. aureus through a lipid peroxidation mechanism whereby AA is oxidized to reactive electrophiles that modify S. aureus macromolecules, eliciting toxicity. This process is rescued by cotreatment with antioxidants as well as in a S. aureus strain genetically inactivated for lcpA (USA300 ΔlcpA mutant) that produces lower levels of reactive oxygen species. However, resistance to AA stress in the USA300 ΔlcpA mutant comes at a cost, making the mutant more susceptible to β-lactam antibiotics and attenuated for pathogenesis in a murine infection model compared to the parental methicillin-resistant S. aureus (MRSA) strain, indicating that resistance to AA toxicity increases susceptibility to other stressors encountered during infection. This report defines the mechanism by which AA is toxic to S. aureus and identifies lipid peroxidation as a pathway that can be modulated for the development of future therapeutics to treat S. aureus infections.IMPORTANCE Despite the ability of the human immune system to generate a plethora of molecules to control Staphylococcus aureus infections, S. aureus is among the pathogens with the greatest impact on human health. One class of host molecules toxic to S. aureus consists of polyunsaturated fatty acids. Here, we investigated the antibacterial properties of arachidonic acid, one of the most abundant polyunsaturated fatty acids in humans, and discovered that the mechanism of toxicity against S. aureus proceeds through lipid peroxidation. A better understanding of the molecular mechanisms by which the immune system kills S. aureus, and by which S. aureus avoids host killing, will enable the optimal design of therapeutics that complement the ability of the vertebrate immune response to eliminate S. aureus infections.


Women's knowledge and attitudes surrounding abortion in Zambia: a cross-sectional survey across three provinces.
In Zambia, despite a relatively liberal legal framework, there remains a substantial burden of unsafe abortion. Many women do not use skilled providers in a well-equipped setting, even where these are available. The aim of this study was to describe women's knowledge of the law relating to abortion and attitudes towards abortion in Zambia. Community-based survey in Central, Copperbelt and Lusaka provinces. 1484 women of reproductive age (15-44 years). Correct knowledge of the legal grounds for abortion, attitudes towards abortion services and the previous abortions of friends, family or other confidants. Descriptive statistics and multivariable logistic regression were used to analyse how knowledge and attitudes varied according to sociodemographic characteristics. Overall, just 16% (95% CI 11% to 21%) of women of reproductive age correctly identified the grounds for which abortion is legal. Only 40% (95% CI 32% to 45% of women of reproductive age knew that abortion was legally permitted in the extreme situation where the pregnancy threatens the life of the mother. Even in urban areas of Lusaka province, only 55% (95% CI 41% to 67%) of women knew that an abortion could legally take place to save the mother's life. Attitudes remain conservative. Women with correct knowledge of abortion law in Zambia tended to have more liberal attitudes towards abortion and access to safe abortion services. Neither correct knowledge of the law nor attitudes towards abortion were associated with knowing someone who previously had an induced abortion. Poor knowledge and conservative attitudes are important obstacles to accessing safe abortion services. Changing knowledge and attitudes can be challenging for policymakers and public health practitioners alike. Zambia could draw on its previous experience in dealing with its large HIV epidemic to learn cross-cutting lessons in effective mass communication on what is a difficult and sensitive issue.


Delays in Diagnosis and Treatment of Bacterial Meningitis in NYC: Retrospective Cohort Analysis.
Community-acquired bacterial meningitis (CABM) morbidity and mortality remains high in those infected. Rapid diagnosis and treatment is paramount to reducing mortality and improving outcome. This retrospective cohort study aims to assess the time from presentation to diagnosis and treatment of vaccine preventable CABM as well as identify possible factors associated with delays in diagnosis and antibiotic administration. A retrospective chart review was conducted of individuals who presented to Columbia University Irving Medical Center (CUIMC), Children's Hospital of New York (CHONY), Mount Sinai Medical Center, and Weill Cornell Medical Center with BM due to Haemophilus influenzae type B, Streptococcus pneumoniae, and Neisseria meningitidis between January 1, 2012 and December 31, 2017. Diagnosis was delayed by more than 8 hours in 13 patients (36.1%) and 5 individuals (13.9%) had a delay of 4 hours or more from presentation to the administration of antibiotics with appropriate CNS coverage. All of these patients were also initially misdiagnosed at an outpatient clinic, outside hospital, or emergency department. This retrospective study identified febrile and/or viral infections not otherwise specified and otitis media as the most common misdiagnoses underlying delays from presentation to diagnosis and to antibiotic treatment in those with BM.


The interaction of triethyltin with components of animal tissues.
1. The distribution of triethyl[(113)Sn]tin chloride in the rat, guinea pig and hamster is not uniform, the highest concentrations being in rat blood and the liver of all three species. 2. Subcellular fractionation of rat liver, brain and kidney shows that triethyltin binds to all fractions to different extents. In the liver of the rat and guinea pig the supernatant fraction contains the largest amount and the highest specific concentration; this triethyltin is bound to a non-diffusible component. 3. Rat haemoglobin is responsible for the binding of triethyltin in rat blood (2 moles of triethyltin/mole of haemoglobin). Haemoglobins from other species have much less affinity for triethyltin. 4. A variety of other proteins do not bind triethyltin.


Rheology and pressurised gyration of starch and starch-loaded poly(ethylene oxide).
This work investigates the rheology and spinning of starch and starch-loaded poly(ethylene oxide) (PEO) by pressurised gyration in order to prepare nanofibres. The spinning dope's rheological properties played a crucial role in fibre formation. Newtonian behaviour is observed in 1-20 wt% starch suspensions and non-Newtonian behaviour is found in all the PEO-starch mixtures. Pressurised gyration of the starch suspensions produced beads only but PEO-starch mixtures generated fibres. The fibre diameter of the PEO-starch samples is shown to be a function of polymer concentration and rotating speed of the gyration system. Fibre formation can only be facilitated below a certain working pressure. The concentration of starch in the PEO-starch mixtures is crucial in defining whether beaded or continuous fibres were generated and this is related to the composition of the spinning dope. FT-IR, XRD and microscopy studies indicated very good miscibility of starch and PEO in the nanofibres. The storage modulus of the PEO-starch were also studied as a function of temperature (30-150°C) and showed interesting results but it was not possible to deduce general trends valid for the entire temperature range.


Does Silver in Different Forms Affect Bacterial Susceptibility and Resistance? A Mechanistic Perspective.
The exceptional increase in antibiotic resistance in past decades motivated the scientific community to use silver as a potential antibacterial agent. However, due to its unknown antibacterial mechanism and the pattern of bacterial resistance to silver species, it has not been revolutionized in the health sector. This study deciphers mechanistic aspects of silver species, i.e., ions and lysozyme-coated silver nanoparticles (L-Ag NPs), against E. coli K12 through RNA sequencing analysis. The obtained results support the reservoir nature of nanoparticles for the controlled release of silver ions into bacteria. This study differentiates between the antibacterial mechanism of silver species by discussing the pathway of their entry in bacteria, sequence of events inside cells, and response of bacteria to overcome silver stress. Controlled release of ions from L-Ag NPs not only reduces bacterial growth but also reduces the likelihood of resistance development. Conversely, direct exposure of silver ions, leads to rapid activation of the bacterial defense system leading to development of resistance against silver ions, like the well-known antibiotic resistance problem. These findings provide valuable insight on the mechanism of silver resistance and antibacterial strategies deployed by E. coli K12, which could be a potential target for the generation of aim-based and effective nanoantibiotics.


Flaxseed improves lipid profile without altering biomarkers of bone metabolism in postmenopausal women.
The risk of cardiovascular disease and osteoporosis drastically increases at the onset of menopause. Phytoestrogens have been suggested to inhibit bone loss and protect the cardiovascular system, in part by improving lipid profiles. The purpose of the present study was to examine the effects of flaxseed, a rich source of the phytoestrogens called lignans, on lipid metabolism and biomarkers of bone turnover in postmenopausal women. Postmenopausal women who were not on hormone replacement therapy were assigned to one of two treatment groups in a double-blind randomized study. Women were asked to consume 40 g of either ground flaxseed or wheat-based comparative control regimen daily for 3 months. In addition, all subjects received 1,000 mg calcium and 400 IU vitamin D daily. Flaxseed supplementation lowered (P < 0.05) both serum total cholesterol and non-high-density lipoprotein cholesterol by 6%, whereas the comparative control regimen had no such effect. Flaxseed regimen reduced serum levels of both low-density- and high-density-lipoprotein cholesterol by 4.7% and triglyceride by 12.8%, albeit not statistically significant. Serum apolipoprotein A-1 and apolipoprotein B concentrations were significantly (P < 0.005) reduced by 6 and 7.5%, respectively, by the flaxseed regimen. Markers of bone formation and resorption were not affected by either of the treatments. The findings of this study indicate that flaxseed supplementation improves lipid profiles but has no effect on biomarkers of bone metabolism in postmenopausal women.


UV-Curable Biobased Polyacrylates Based on a Multifunctional Monomer Derived from Furfural.
The controlled polymerization of a new biobased monomer, 4-oxocyclopent-2-en-1-yl acrylate (4CPA), was established via reversible addition-fragmentation chain transfer (RAFT) (co)polymerization to yield polymers bearing pendent cyclopentenone units. 4CPA contains two reactive functionalities, namely, a vinyl group and an internal double bond, and is an unsymmetrical monomer. Therefore, competition between the internal double bond and the vinyl group eventually leads to gel formation. With RAFT polymerization, when aiming for a degree of polymerization (DP) of 100, maximum 4CPA conversions of the vinyl group between 19.0 and 45.2% were obtained without gel formation or extensive broadening of the dispersity. When the same conditions were applied in the copolymerization of 4CPA with lauryl acrylate (LA), methyl acrylate (MA), and isobornyl acrylate, 4CPA conversions of the vinyl group between 63 and 95% were reached. The additional functionality of 4CPA in copolymers was demonstrated by model studies with 4-oxocyclopent-2-en-1-yl acetate (1), which readily dimerized under UV light via [2 + 2] photocyclodimerization. First-principles quantum mechanical simulations supported the experimental observations made in NMR. Based on the calculated energetics and chemical shifts, a mixture of head-to-head and head-to-tail dimers of (1) were identified. Using the dimerization mechanism, solvent-cast LA and MA copolymers containing 30 mol % 4CPA were cross-linked under UV light to obtain thin films. The cross-linked films were characterized by dynamic scanning calorimetry, dynamic mechanical analysis, IR, and swelling experiments. This is the first case where 4CPA is described as a monomer for functional biobased polymers that can undergo additional UV curing via photodimerization.


Structure-based calculation of the equilibrium folding pathway of proteins. Correlation with hydrogen exchange protection factors.
A new statistical thermodynamic formalism has been developed in order to describe the equilibrium folding pathway of proteins. The resulting formalism allows calculation of the probabilities that individual amino acid residues will be in a native or native-like conformation for any given degree of folding of the protein molecule. The residue probabilities are defined by the probability distribution of conformational states and can be used to calculate experimental quantities like native-state, hydrogen exchange protection factors. A combinatorial algorithm aimed at generating a large ensemble of conformational states (10(4) to 10(6)) using the native structure as a template has been developed. The Gibbs energy and corresponding probability of each conformational state is estimated by using a previously developed structural parametrization of the energetics. The approach has been applied to five different proteins: hen egg-white lysozyme, equine lysozyme, bovine pancreatic trypsin inhibitor, staphylococcal nuclease and turkey ovomucoid third domain. The validity of the approach has been tested by comparing predicted and experimental hydrogen exchange protection factors. It is shown that for the above proteins 76%, 73%, 74%, 78% and 81% of all observed protection factors are predicted correctly. Furthermore, on average, the magnitude of the predicted protection factors, expressed as apparent free energies per residue deviate less than 1 kcal/mol from those obtained experimentally. These results represent the first attempt at predicting both the location and magnitude of hydrogen exchange protection factors from the high-resolution structure of a protein. The good agreement between experimental and predicted values has permitted a close examination of the nature of the equilibrium folding intermediates existing under conditions of maximal stability of the native state.


Optical regenerative NRZ to RZ format conversion based on cascaded lithium niobate modulators.
Optical regenerative nonreturn-to-zero (NRZ) to return-to-zero (RZ) format conversion using a lithium niobate phase modulator and a lithium niobate intensity modulator is proposed and demonstrated. The key advantage of the proposed format converter is that the converted RZ signal has a very small pulse width, which can be multiplexed to a higher bit rate using optical time division multiplexing technology. The operation can greatly reduce the timing jitter of the degraded NRZ signal due to the regenerative property of the proposed scheme. Besides, the format converter can also support multi-channel operation. An experiment is performed with the feasibility of the scheme demonstrated.


Development of a thiol-ene based screening platform for enzyme immobilization demonstrated using horseradish peroxidase.
Efficient immobilization of enzymes on support surfaces requires an exact match between the surface chemistry and the specific enzyme. A successful match would normally be identified through time consuming screening of conventional resins in multiple experiments testing individual immobilization strategies. In this study we present a versatile strategy that largely expands the number of possible surface functionalities for enzyme immobilization in a single, generic platform. The combination of many individual surface chemistries and thus immobilization methods in one modular system permits faster and more efficient screening, which we believe will result in a higher chance of discovery of optimal surface/enzyme interactions. The proposed system consists of a thiol-functional microplate prepared through fast photochemical curing of an off-stoichiometric thiol-ene (OSTE) mixture. Surface functionalization by thiol-ene chemistry (TEC) resulted in the formation of a functional monolayer in each well, whereas, polymer surface grafts were introduced through surface chain transfer free radical polymerization (SCT-FRP). Enzyme immobilization on the modified surfaces was evaluated by using a rhodamine labeled horseradish peroxidase (Rho-HRP) as a model enzyme, and the amount of immobilized enzyme was qualitatively assessed by fluorescence intensity (FI) measurements. Subsequently, Rho-HRP activity was measured directly on the surface. The broad range of utilized surface chemistries permits direct correlation of enzymatic activity to the surface functionality and improves the determination of promising enzyme-surface candidates. The results underline the high potential of this system as a screening platform for synergistic immobilization of enzymes onto thiol-ene polymer surfaces. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1267-1277, 2017.


Supplying emergency contraception via community pharmacies in the UK: reflections on the experiences of users and providers.
This paper discusses findings from an evaluation of a scheme to provide free emergency hormonal contraception (EHC) via community pharmacies in the North-West of England. Drawing on interview data with pharmacists taking part in the scheme and focus groups with users, we tentatively suggest that the scheme was largely well received. The benefits of the service, cited by both pharmacists and users, included enhanced access to EHC, at times when it was needed, and at no cost to the user. In particular, users noted a welcome absence of judgmental attitudes when accessing the service. Pharmacists too were positive about the service, not least because they believed that it conferred enhanced professional status. However, both users and pharmacists had a number of major concerns about the schemes, centring on the potential for misuse, changes in contraceptive behaviour and the impact on sexually transmitted infections. We conclude that more research is needed to explore these issues.


Short- and long-term residents of nursing homes.
This article analyzes reported data on length of stay of discharged patients from the 1977 National Nursing Home Survey. Assuming that patients admitted to nursing homes are one of two types, short-stayers and long-stayers, the statistically best-fitting proportions and expected lengths of stay for the two types are derived. The results are applied to statistics on characteristics of resident and discharged patients to find admission characteristics that differentiate the two groups. Because long-stayers and short-stayers have quite different characteristics, nursing home statistics will be very different if one bases them on discharges or a cross-section of residents. For example, we estimate that short-stay patients represent 61 per cent of discharges but only 9 per cent of residents. A further application shows that, including deaths in hospitals, almost half of discharged patients die, rather than the usually cited figure of 25 per cent.


[Some aspects of professional activities and a risk to the health status of aircraft designers].
Morbidity associated with temporary disability and risks for major syndromes have been studied in aircraft designers who work under the conditions regarded as hazardous (the third degree of Class 2). The risks for major syndromes have been analyzed in relation to sociopsychological characteristics. The manifestations of the emotional exhaustion syndrome have been examined in the workers exposed to industrial stresses.


Cigarette Smoking Attenuates Kidney Protection by Angiotensin-Converting Enzyme Inhibition in Nondiabetic Chronic Kidney Disease.
Cigarette smoking exacerbates the estimated glomerular filtration rate (eGFR) decline in nondiabetic chronic kidney disease (CKD) despite the kidney protection that is achieved by angiotensin converting enzyme inhibition (ACEI). Whether smoking cessation restores ACEI-related kidney protection is not known. This 5-year, prospective, prevention trial recruited 108 smokers and 108 nonsmokers with stage-2 nondiabetic CKD with primary hypertension and urine albumin-to-creatinine ratio (Ualb) >200 mg/g. All smokers underwent smoking cessation intervention programs. Blood pressure was reduced in all participants toward achieving a goal of <130 mm Hg with regimens including ACEI. The primary outcome was eGFR change, and secondary outcomes included Ualb and urine levels of angiotensinogen (UATG), a surrogate for kidney angiotensin II (AII) levels, and isoprostane 8-isoprostaglandin F2α (U8-iso), an indicator of oxidative stress. One-year Ualb was lower than baseline in nonsmokers but not in either smoking group, supporting greater ACEI-related kidney protection in nonsmokers than smokers. Higher Ualb at 1 year in continued smokers was associated with higher UATG and higher U8-iso, consistent with smoking-induced AII and increased oxidative stress contributing to less ACEI-related kidney protection in smokers. Baseline eGFR was not different among groups (p = 0.92), but 5-year eGFR was higher in quitters than in continued smokers (62.0 ± 5.4 vs. 52.9 ± 5.6 mL/min/1.73 m2, p < 0.001); this value was lower in quitters than in nonsmokers (64.7 ± 5.6 mL/min/1.73 m2, p = 0.02). Smoking cessation compared with continued smoking ameliorates eGFR decline in nondiabetic CKD treated with ACEI, possibly by restoring kidney-protective effects of ACEI through reductions in kidney AII and oxidative stress.


Tubal flushing for subfertility.
A possible therapeutic effect of diagnostic tubal patency testing has been debated in the literature for half a century. Further debate surrounds whether oil-soluble or water-soluble contrast media might have the bigger fertility-enhancing effect. To evaluate the effect of flushing a woman's fallopian tubes with oil- or water-soluble contrast media on subsequent fertility outcomes in couples with infertility. We searched the Cochrane Menstrual Disorders and Subfertility Group's specialised register of trials (searched 31 January 2007), MEDLINE, EMBASE, Biological Abstract and reference lists of articles. All randomised trials comparing tubal flushing with oil-soluble contrast media or tubal flushing with water-soluble media or with no treatment in women with subfertility. Four authors independently assessed trial quality and extracted data. We contacted study authors for additional information. We collected adverse effects information from the trials. Twelve trials involving 2079 participants were included. Tubal flushing with oil-soluble media versus no intervention was associated with a significant increase in the odds of live birth (Peto OR 2.98, 95% CI 1.40 to 6.37) and of pregnancy (Peto OR 3.30, 95% CI 2.00 to 5.43). For the comparison of tubal flushing with oil-soluble media versus tubal flushing with water-soluble media, the increase in the odds of live birth for tubal flushing with oil-soluble versus water-soluble media (Peto OR 1.49, 95% CI 1.05 to 2.11) was based on two trials where statistical heterogeneity was present and the higher quality trial showed no significant difference; there was no evidence of a significant difference in the odds of pregnancy (Peto OR 1.21, 95% CI 0.95 to 1.54). The addition of oil-soluble media to flushing with water-soluble media showed no evidence of a significant difference in the odds of pregnancy (Peto OR 1.28, 95% CI 0.92 to 1.79) or live birth (Peto OR 1.06, 95% CI 0.64 to 1.77). There were no serious adverse event reported. There is evidence of effectiveness of tubal flushing with oil-soluble contrast media in increasing the odds of pregnancy and live birth versus no intervention. Future robust randomised trials, comparing oil-soluble versus water-soluble media, water-soluble media versus no intervention and tubal flushing versus established treatments for infertility would be a useful further guide to clinical practice.


Renal and vascular effects of chronic nitric oxide synthase inhibition: involvement of endothelin 1 and angiotensin II.
Endothelin 1 (ET-1) is a potent vasoconstrictor implicated in the control of blood pressure and renal function. Its effects can be modulated by nitric oxide (NO), which inhibits ET-1 production and action. Recently, we reported that ET-1 production can also be modulated by angiotensin II (AngII) in vivo. To investigate the interactions between NO, ET-1, and AngII in hypertension and renal dysfunction, we assessed immunoreactive ET-1 (ir-ET-1) concentration in plasma and urine as well as in vascular and renal tissues of rats with chronic inhibition of NO synthesis, in the presence and the absence of the AngII type 1 receptor antagonist losartan. Normal (protocols A and B) and uninephrectomized rats (protocol C) received the L-arginine analog N(G)-nitro-L-arginine methyl ester (L-NAME), an inhibitor of NO synthesis, 0.05% (protocol A) or 0.1% (protocols B and C), with or without losartan (20 mg x kg(-1) x day(-1)). After 6 weeks, systolic blood pressure was significantly increased in L-NAME rats compared with the controls (p < 0.01), while serum creatinine and urea, creatinine clearance, and proteinuria were similar to control values. However, ir-ET-1 concentration in plasma and in the thoracic aorta was augmented in animals receiving 0.1% L-NAME (1 < 0.01), while it was unchanged in the mesenteric arterial bed, preglomerular arteries, and glomeruli. In contrast, ir-ET-1 concentration was decreased in the renal papilla (p < 0.05) as well as in the urine of L-NAME rats (p < 0.01). Treatment with losartan significantly attenuated the rise in systolic blood pressure induced by L-NAME (p < 0.01). Losartan also normalized the increased ir-ET-1 concentration in plasma and in the thoracic aorta, but had no effect on tissues with normal or reduced ir-ET-1 levels. These results indicate that chronic inhibition of NO synthase with L-NAME induces hypertension without renal dysfunction. Increased ET-1 production in some blood vessels and elevated circulating ET-1 concentration may contribute to the maintenance of high blood pressure. The reduction of systolic blood pressure by losartan supports a role for AngII in the pathogenesis of this form of hypertension, which may be due, at least in part, to the modulation of ET-1 production.


Chemically Robust Antifog Nanocoating through Multilayer Deposition of Silica Composite Nanofilms.
A coating must remain intact to perform its inherent functions on a surface, and often functional organic coatings fail due to deterioration because of their intrinsic vulnerabilities. In this work, we present a biomimetic material based on a glass sponge to provide a robust silica composite nanocoating with an antifog effect. The silica composite nanocoating was constructed with a binary film structure consisting of (1) a Fe(III)-tannic acid (TA) nanofilm for adhesion to coat the substrates and (2) a SiO2 layer to enhance the durability of the coating. Due to the universal coating property of Fe(III)-TA nanofilms, we demonstrated that the silica composite nanocoating was effective regardless of the substrate. By layer-by-layer assembly of the silica composite, it is possible to precisely control the nanocoating thickness. The superhydrophilic nature of the SiO2 layer showed an exceptional antifog effect that remained intact against multiple deteriorative conditions, including acid treatment, peroxide degradation, sudden temperature change, severe heat conduction, and oil contamination. In addition, the silica composite nanocoating is scalable for surfaces of different shapes and sizes with the aid of a spray-assisted deposition technique. The bioinspired, multicomposite nanocoating strategy herein contributes to the improvement of organic coatings for uses in applications to tackle current technological problems.


Effects of crude protein content on intake and digestion of coastal bermudagrass hay by horses.
This study was conducted to determine the effects of forage crude protein (CP) level on intake and digestion of Coastal bermudagrass hay by horses. Four cecally fistulated geldings were used in a 4 × 4 Latin square design with four treatments and four periods. Horses were fed one of four Coastal bermudagrass hays consisting of 7%, 10%, 13%, or 16% CP during each of the four 15-d periods. Intake and apparent digestibility were determined for each horse at the end of each period by total fecal collection. In addition, cecal fluid and blood samples were collected on the last day of each period for the determination of cecal ammonia, cecal pH, plasma urea nitrogen (PUN), and plasma glucose concentrations. Data were analyzed using PROC MIXED of SAS. CP concentration of Coastal bermudagrass hay influenced equine intake and digestion. Increasing CP concentration linearly increased digestible organic matter intake (DOMI) from 3.79 to 5.98 kg/d for 7% and 16% CP hay, respectively (P = 0.04). Furthermore, as the forage CP level increased, CP intake increased linearly (P < 0.01). The forage CP level had no effect on forage dry matter intake. Quadratic effects (P ≤ 0.05) were observed for forage OM, neutral detergent fiber, acid detergent fiber, and digestible energy. Overall digestibility was lowest for the 7% CP hay and highest for the 10% CP hay. Cecal pH remained above 6.62 irrespective of treatment and time, indicating that cecal pH was suitable for microbial growth. As the forage CP level increased, cecal ammonia concentration increased linearly from 0.03 mM for the 7% to 1.74 mM for the 16% CP hay (P < 0.01). Concentration of plasma glucose also linearly increased (P = 0.04) from 68.77 to 73.68 mg/dL as CP concentration increased from 7% to 16% CP. PUN exhibited a quadratic effect as concentration increased (P < 0.01) from 4.34 to 5.61 mM for the 7% and 16% CP hays, respectively. Overall, the 10% CP hay had the highest digestibility due to its higher OM digestion. As forage OMI increased, digestible OM increased until physiological capacity for digestion is exceeded. At that point, digestion will decline with the decrease in OMI, explaining the lower digestion for other forages fed.


Is everyone hot in the city? Spatial pattern of land surface temperatures, land cover and neighborhood socioeconomic characteristics in Baltimore, MD.
Urban heat island effect refers to the phenomenon that ambient air and surface temperatures in urban areas are several degrees higher than surrounding rural areas. Higher temperatures not only impact the comfort of urban dwellers, but also increase energy use, ozone production, and the risk of death for humans in a heat wave. Our research focuses on the variation in land surface temperature in the Gywnns Fall Watershed, Maryland. We found that land surface temperature is highly variable spatially, resulting in "hotspots" within the heat island. We further explore how this temperature variation relates to social factors on the scale of the census-based block group. We show that land surface temperature is statistically higher in block groups that are characterized by low income, high poverty, less education, more ethnic minorities, more elderly people and greater risk of crime. These variables were mapped to evaluate the spatial relationship of land surface temperatures to social factors. This spatially explicit approach facilitates identification of specific areas to prioritize for heat prevention and intervention efforts. We demonstrate, through an exercise, how incorporating data on land surface temperature and social factors into heat intervention strategies could contribute to efficient allocation of limited resources and services. The exercise also indicates where heat prevention efforts, such as tree-planting programs, are most needed to help reduce heat exposure and moderate the urban heat island effect.


Macrophage innate training induced by IL-4 and IL-13 activation enhances OXPHOS driven anti-mycobacterial responses.
Macrophages are a highly adaptive population of innate immune cells. Polarization with IFNγ and LPS into the 'classically activated' M1 macrophage enhances pro-inflammatory and microbicidal responses, important for eradicating bacteria such as Mycobacterium tuberculosis. By contrast, 'alternatively activated' M2 macrophages, polarized with IL-4, oppose bactericidal mechanisms and allow mycobacterial growth. These activation states are accompanied by distinct metabolic profiles, where M1 macrophages favor near exclusive use of glycolysis, whereas M2 macrophages up-regulate oxidative phosphorylation (OXPHOS). Here, we demonstrate that activation with IL-4 and IL-13 counterintuitively induces protective innate memory against mycobacterial challenge. In human and murine models, prior activation with IL-4/13 enhances pro-inflammatory cytokine secretion in response to a secondary stimulation with mycobacterial ligands. In our murine model, enhanced killing capacity is also demonstrated. Despite this switch in phenotype, IL-4/13 trained murine macrophages do not demonstrate M1-typical metabolism, instead retaining heightened use of OXPHOS. Moreover, inhibition of OXPHOS with oligomycin, 2-deoxy glucose or BPTES all impeded heightened pro-inflammatory cytokine responses from IL-4/13 trained macrophages. Lastly, this work identifies that IL-10 attenuates protective IL-4/13 training, impeding pro-inflammatory and bactericidal mechanisms. In summary, this work provides new and unexpected insight into alternative macrophage activation states in the context of mycobacterial infection.


How the humanities shape medical culture: Knowing Wegener and other Nazi eponyms.
This article was migrated. The article was marked as recommended. While the medical humanities have experienced a renaissance, they are still largely a peripheral component of medical education. This is troublesome because the humanities include a number of disciplines that are foundational in understanding medicine and how it should be practiced. Nonetheless, current medical culture makes it difficult to fully incorporate the humanities into curriculum. We therefore propose an incremental approach to shaping the medical culture that can easily be incorporated into daily teaching as opposed to designing additional classes and resources that must be added to existing educational structures. An example of this approach is reviewed here through teaching historical and ethical lessons surrounding Nazi eponyms. The use of names like Wegener provide brief opportunities for sidebars during clinical lectures to remind learners that empirical data do not provide ethical direction and that our medical history has included atrocities that remind us to practice conscientiously. We provide other examples that can be included in daily learning. This approach eschews the burdens associated with large curricular changes, such as student resistance/apathy and logistical barriers, and can be easily implemented. It also enables change to be gradual and through structures that have already been established, allowing learners to see the benefits of insights from the humanities in small, digestible segments. Through this approach, medical culture can be shaped towards a greater appreciation toward the medical humanities.


Heart rate variability as a biomarker for autonomic nervous system response differences between children with chronic pain and healthy control children.
Studies in adults have demonstrated a relationship between lowered heart rate variability (HRV) and poor health. However, less is known about the role of autonomic arousal in children's well-being. The aim of the current study was to examine resting HRV in children with chronic pain compared to healthy control children and, further, to examine children's HRV following a series of acute experimental pain tasks in both groups. Participants included 104 healthy control children and 48 children with chronic pain aged 8-17 years. The laboratory session involved a 5-minute baseline electrocardiogram followed by four pain induction tasks: evoked pressure, cold pressor, focal pressure, and a conditioned pain modulation task. After the tasks were complete, a 5-minute post-task electrocardiogram recording was taken. Spectral analysis was used to capture high-frequency normalized power and the ratio of low-to-high frequency band power, signifying cardiac vagal tone and sympathetic balance, respectively. Results revealed that children with chronic pain had significantly lower resting HRV (signified by low high-frequency normalized power and high ratio of low-to-high frequency band power) compared to healthy children; moreover, a significant interaction between groups and time revealed that children with chronic pain displayed a static HRV response to the pain session compared to healthy children, whose HRV was reduced concomitant with the pain session. These findings suggest that children with chronic pain may have a sustained stress response with minimal variability in response to new acute pain stressors.


Understanding the electronic structures of graphene quantum dot physisorption and chemisorption onto the TiO2 (110) surface: a first-principles calculation.
We investigated the interfacial electronic structure and charge transfer properties of graphene quantum dot (GQD) physisorption and chemisorption on the TiO(2) (110) surface from density functional theory calculations. The simulations show that a slight charge transfer occurs in physisorption case while a significant charge transfer takes place in chemisorption configuration. We present a detailed comparison of the similarities and differences between the electronic structures. The similarities originate from the positive work function difference in both the physisorption and chemisorption configurations, which is able to drive electron transfer from GQD into TiO(2), leading to charge separation across the GQD-TiO(2) interface. The differences stem from the interaction between the GQD and TiO(2) substrate. For example, GQD bounds to TiO(2) surface through van der Waals interactions in the case of physisorption. In the chemisorption configuration, however, there exists strong covalent bonding between them. This leads to much more efficient charge separation for chemisorption than for physisorption. Furthermore, the GQD-TiO(2) composites show large band-gap narrowing that could extend the optical absorption edge into the visible-light region. This should imply that chemisorbed GQDs produce a composite with better photocatalytic and photovoltaic performance than composites formed through physisorption.


Nocturia compounds nocturnal wakefulness in older individuals with insomnia.
To determine the impact of nocturia on objective measures of sleep in older individuals with insomnia. The sleep and toileting patterns of a group of community-dwelling older men (n = 55, aged 64.3 ± 7.52 years) and women (n = 92, aged 62.5 ± 6.73 years) with insomnia were studied for two weeks using sleep logs and one week using actigraphy. The relationships between nocturia and various sleep parameters were analyzed with ANOVA and linear regression. More than half (54.2% ± 39.9%) of all log-reported nocturnal awakenings were associated with nocturia. A greater number of trips to the toilet was associated with worse log-reported restedness (p < 0.01) and sleep efficiency (p < 0.001), as well as increases in actigraph-derived measures of the number and length of nocturnal wake bouts (p < 0.001) and wake after sleep onset (p < 0.001). Actigraph-determined wake bouts were 11.5% ± 23.5% longer on nights on which there was a trip to the toilet and wake after sleep onset was 20.8% ± 33.0% longer during these nights. Nocturia is a common occurrence in older individuals with insomnia and is significantly associated with increased nocturnal wakefulness and decreased subjective restedness after sleep.


Usefulness and Validity of Continuous Performance Tests in the Diagnosis of Attention-Deficit Hyperactivity Disorder Children.
Despite the popularity of continuous performance tests (CPT) in supporting the diagnostic procedure of attention-deficit hyperactivity disorder (ADHD), these measures are still controversial mainly due to limited sensitivity, specificity, and ecological validity. Thus, there continues to be a need for further validation of these objective attention measures. The purpose of this study was to evaluate the usefulness of a CPT that includes environmental distracting stimuli, in supporting the diagnosis of ADHD in children. Participants were 798 children aged 7-12 years (493 boys and 305 girls). The ADHD group included 339 children, whereas the control group included 459 children without ADHD. The study employed the MOXO-CPT, which incorporates visual and auditory stimuli serving as environmental distractors. Compared to their unaffected peers, children with ADHD received significantly lower scores in all 4 CPT indices: attention, timing, hyperactivity, and impulsivity. Specifically, ADHD children were less attended to the stimuli and performed fewer reactions on accurate timing. Furthermore, children with ADHD performed significantly more impulsive and hyperactive responses than controls. Receiver operating characteristic analysis revealed fair to excellent diagnostic ability of all CPT indices except impulsivity, which showed poor ability to distinguish ADHD children from controls. The test's total score yielded excellent diagnostic performance. MOXO-CPT consistently distinguished between children with ADHD and their unaffected peers, so that children with ADHD performed worse than controls in all study indices. Integration of CPT indices improves the diagnostic capacity of ADHD and may better reflect the complexity and heterogeneity of ADHD.


The goat (Capra hircus) mammary gland secretory tissue proteome as influenced by weight loss: A study using label free proteomics.
Seasonal weight loss (SWL) is a significant limitation to animal production. Breeds that have evolved in harsh climates have acquired tolerance to SWL through selection. Herein, label free proteomics was used to characterize the effects of SWL in two goat breeds with different levels of adaptation to nutritional stress: Majorera (tolerant) and Palmera (susceptible). Nine Majorera and 10 Palmera dairy goats were used, divided in 4 sets, 2 for each breed: underfed and a control group: Majorera Control (MC), Palmera Control (PC), Majorera Restricted (ME) and Palmera Restricted (PE). At day 22, mammary gland biopsy samples were obtained. Label free proteomic analysis enabled the identification of over 1000 proteins, of which 96 showed differential expression between two of the groups within studied comparisons. Majorera breed showed higher expression of immune system related proteins. In contrast, Palmera breed showed higher expression of proteins related to apoptosis. Results indicate that the two goat breeds have a distinct metabolism reaction to SWL, and that proteins related to the immune system and apoptosis such as cadherin-13, collagen alpha-1, nidogen-2, clusterin and protein s100-A8 could be considered putative candidates as markers of tolerance to SWL. Seasonal weight loss (SWL) is one of the major constraints to animal production in the tropics. We compared the proteomics profiles of two dairy goat breeds with different levels of tolerance to SWL under control and feed restriction conditions using label free proteomics. We have identified over 1000 proteins in the goat mammary gland, of which 96 showed differential expression. Despite the fact that both breeds showed a decrease in the number of proteins related to protein, carbohydrates and fat biosynthesis, the tolerant breed showed higher expression of immune system related proteins compared to the susceptible breed. On the contrary, the susceptible breed had higher expression of apoptosis related proteins indicating that both breeds reacted differently to weight loss and that proteins related to the immune system and apoptosis such as cadherin-13, collagen alpha 1, nidogen-2 or clusterin may be suggested as markers of tolerance to SWL.


Differential roles of NMDA Receptor Subtypes NR2A and NR2B in dendritic branch development and requirement of RasGRF1.
N-methyl-D-aspartate receptors (NMDARs) are known to regulate axonal refinement and dendritic branching. However, because NMDARs are abundantly present as tri-heteromers (e.g., NR1/NR2A/NR2B) during development, the precise role of the individual subunits NR2A and NR2B in these processes has not been elucidated. Ventral spinal cord neurons (VSCNs) provide a unique opportunity to address this problem, because the expression of both NR2A and NR2B (but not NR1) is downregulated in culture. Exogenous NR2A or NR2B were introduced into these naturally NR2-null neurons at 4 DIV, and electrophysiological recordings at 11 DIV confirmed that synaptic NR1NR2A receptors and NR1NR2B receptors were formed, respectively. Analysis of the dendritic architecture showed that introduction of NR2B, but not NR2A, dramatically increased the number of secondary and tertiary dendritic branches of VSCNs. Whole cell patch-clamp recordings further indicated that the newly formed branches in NR2B-expressing neurons were able to establish functional synapses because the frequency of miniature AMPA-receptor synaptic currents was increased. Using previously described mutants, we also found that disruption of the interaction between NR2B and RasGRF1 dramatically impaired dendritic branch formation in VSCNs. The differential role of the NR2A and NR2B subunits and the requirement for RasGRF1 in regulating branch formation was corroborated in hippocampal cultures. We conclude that the association between NR1NR2B-receptors and RasGRF1 is needed for dendritic branch formation in VSCNs and hippocampal neurons in vitro. The dominated NR2A expression and the limited interactions of this subunit with the signaling protein RasGRF1 may contribute to the restricted dendritic arbor development in the adult CNS.


Opioid growth factor regulates the cell cycle of human neoplasias.
The native opioid growth factor (OGF), [Met5]-enkephalin, is a tonic inhibitory peptide that modulates cell proliferation and migration, as well as tissue organization, during development, cancer, homeostatic cellular renewal, wound healing, and angiogenesis. OGF action is mediated by the OGF receptor (OGFr). To investigate the target of OGF as to cell proliferation, the effects of excess OGF, and a deprivation of OGF-OGFr interaction by an opioid antagonist, naltrexone (NTX), were examined in 3 human cancer cell lines: pancreatic (BxPC-3), colon (HT-29), and head and neck (CAL-27). OGF exposure decreased growth, DNA synthesis, and mitosis, and increased the doubling time from control levels. FACS analysis revealed a marked increase in cells in the G0/G1 phase and compensatory reduction in cells in S and G2/M phases. Consistent with this observation, the percentage of labeled mitosis (PLM) analysis showed a notable increase in the time of the G0/G1 phase. Receptor blockade with NTX increased the rate of growth, length of DNA synthesis and mitotic phases, and decreased doubling time from control values. FACS analysis indicated an increase in the proportion of cells in S and G2/M phases, and a decrease in the number of cells in the G0/G1 phase. PLM evaluation demonstrated a shortening of the length of the S and G2 phases in the 3 cell lines, and decreases in the M and G0/G1 phases in some cancers. These results indicate that OGF action is directed at the G0/G1 phase, but interruption of OGF-OGFr interfacing has widespread repercussions on the cell cycle. The data on blockade of OGF-OGFr during log phase growth suggest a requisite escorting of the growth peptide and its receptor through the cell cycle.


Heat-induced thermal tolerance and radiation resistance to apoptosis in human lymphocytes.
We have investigated heat- and radiation-induced apoptosis in human lymphocytes in vitro. We have previously shown that apoptosis was induced by radiation at doses as low as 0.05 Gy. Here we report that heat induced apoptosis in human lymphocytes in a temperature- and time-dependent manner. Temperatures at or below 42 degrees C, for up to 90 min, did not cause lymphocytes to undergo apoptosis, whereas temperatures at or above 43 degrees C, for 30 min and longer, did induce apoptosis. Lymphocytes were protected against apoptosis induced by 44 degrees C heat by a prior heat shock of 42 degrees C for 30 min. Heat-induced thermal tolerance developed immediately following the inducing heat shock, was greater after 4 h, and persisted for at least 24 h. While heat also induced radiation resistance, this change was minor and not apparent until about 24 h after the heat shock. Prior to the development of radiation resistance, heat shock sensitized lymphocytes to radiation-induced apoptosis. We have previously shown that radiation-induced apoptosis in lymphocytes varies between donors and therefore may be useful in assessing individual radiosensitivity. We report here that heat also induced variable levels of apoptosis in lymphocytes from different donors, although the range of responses was not as large as those observed with radiation-induced apoptosis. In summary, heat shock induces tolerance to heat-induced apoptosis and results first in sensitization and then protection of lymphocytes against radiation-induced apoptosis.


Posterior tibial nerve sensory blockade duration prolonged by adding dexmedetomidine to ropivacaine.
Dexmedetomidine, an α(2)-receptor agonist, prolongs analgesia when used in neuraxial and IV blocks. We evaluated the effect of dexmedetomidine added to ropivacaine for tibial nerve block on the duration of the sensory blockade. For this prospective, randomized, controlled, double-blind, crossover trial, 14 healthy volunteers were allocated to 2 groups. All volunteers received an ultrasound-guided tibial nerve block 4 to 5 cm proximally to the medial malleolus. In group R, 10 mL of 0.5% ropivacaine was injected for the block; in group RD, 10 mL of a solution containing 0.5% ropivacaine with 1 μg/kg of dexmedetomidine was administered. After the injection, monitoring of vital signs, evaluation of onset and resolution of sensory block, and level of sedation (Observer's Assessment of Alertness/Sedation scale) were performed. Three weeks later, the same procedure was repeated, but the study subjects were allocated to the other group in a crossover fashion. The primary end point was the duration of sensory blockade. The time and carryover effects were also evaluated. Secondary outcomes were the onset time and the presence of adverse effects such as hypotension, bradycardia, hypoxia, and sedation. Sensory blocks lasted longer in group RD than in group R (21.5 vs 16.2 hours; mean pairwise difference 5.3 hours [95% confidence interval: 3.9-6.7 hours]; P < 0.0001). Onset times were similar between groups. The mean systolic and diastolic blood pressure levels were stable throughout the study period in group R. In group RD, a noticeable decrease in systolic and diastolic blood pressure was observed between 60 and 480 minutes (P < 0.05); 2 volunteers experienced a 30% decrease in systolic blood pressure when compared with the baseline value as compared with none in group R. Heart rate was similar between groups except at 60 minutes (P < 0.01). Dexmedetomidine added to ropivacaine for tibial nerve block prolongs the duration of sensory blockade with similar onset time. However, patients should be monitored for potential adverse effects such as hypotension, bradycardia, and sedation.


Meeting the challenge of adolescent health care in a group practice HMO.
The Teen and Young Adult Health Center of Kaiser Permanente was designed to meet the challenge of adolescent health care by providing a constellation of services, including routine medical care, as well as services related to sexuality and reproduction, violent behavior, and emotional and mental health. We have made health care available, accessible, and acceptable for teens and young adults. We are evaluating the results of these efforts regarding health, morbidity, and mortality, and would urge other HMOs to explore offering these services to their adolescent and young adult members.


Using oxidized low-density lipoprotein autoantibodies to predict restenosis after balloon angioplasty in patients with acute myocardial infarction.
Oxidized low-density lipoproteins (oxLDL) and oxidized low-density lipoprotein autoantibodies (OLAB) have been detected in human plasma and atherosclerotic lesions. OLAB appear to play a role in the clearance of oxLDL from circulation. Higher levels of OLAB appear to be associated with a reduced risk of a wide range of cardiovascular diseases. We investigated the prognostic value of plasma oxLDL and OLAB in patients undergoing primary coronary balloon angioplasty for acute ST-elevation myocardial infarction (STEMI). Plasma oxLDL and OLAB concentrations were measured in 56 patients with acute STEMI before primary angioplasty, and then 3 days, 7 days and 1 month after the acute event. Follow-up angiography was repeated 6 months later to detect the presence of restensosis (defined as >50% luminal diameter stenosis). The thrombolysis in myocardial infarction (TIMI) risk score was calculated to determine the relationship between OLAB/oxLDL ratio and TIMI risk scores. Of the 56 patients, 18 (31%) had angiographic evidence of restenosis. Plasma OLAB concentrations were significantly lower in the restenosis group before angioplasty (181±114 vs. 335±257 U/L, p = 0.003), and at day 3 (155±92 vs. 277±185 U/L, p<0.001) and day 7 (177±110 vs. 352±279 U/L, p<0.001) after the acute event. There was no difference in oxLDL concentration between the two groups. The ratio of OLAB/oxLDL positively correlated with TIMI risk scores before angioplasty (p for trend analysis, p = 0.004), at day 3 (p = 0.008) and day 7 (p<0.001) after STEMI. A relative deficit of OLAB, and hence likely impaired clearance of oxLDL, is associated with the risk of arterial restenosis after primary angioplasty for acute STEMI.


Loss of granule myeloperoxidase during in vitro culture of human monocytes correlates with decay in antiprotozoa activity.
Human monocytes maintained in culture lose microbicidal activity against intracellular protozoa which has been correlated with attenuation of the respiratory burst. The granule enzyme myeloperoxidase, which can markedly amplify hydrogen peroxide-dependent antimicrobial activity, is also lost in vitro. Adherent monocytes were examined immediately, 3 and 10-14 days following explantation, for the magnitude of the stimulated respiratory burst and for cellular myeloperoxidase. Fresh cells generated 254 +/- 38 nmol O2-/mg protein as compared to a peak of 782 +/- 45 nmol O2-/mg at 3 days and less than 100 nmol O2-/mg after 10-14 days. The myeloperoxidase content of the cells also decreased; over 85% was lost after 3 days. Fresh monocytes killed over 90% of ingested Toxoplasma gondii or Leishmania major. In contrast, 10-14 day explanted monocytes killed only 12% of ingested Toxoplasma and 33% of Leishmania, and surviving organisms replicated readily. The 3-day monocytes were significantly less able to kill protozoa than were fresh cells despite their nearly 3-fold greater generation of O2-. If peroxidase was reintroduced into 3-day monocytes by coating organisms with eosinophil peroxidase prior to phagocytosis, their antiprotozoa activity was nearly restored to that of freshly explanted cells.


Hepatic gene expression in rainbow trout (Oncorhynchus mykiss) exposed to different hydrocarbon mixtures.
Traditional biomarkers for hydrocarbon exposure are not induced by all petroleum substances. The objective of this study was to determine if exposure to a crude oil and different refined oils would generate a common hydrocarbon-specific response in gene expression profiles that could be used as generic biomarkers of hydrocarbon exposure. Juvenile rainbow trout (Oncorhynchus mykiss) were exposed to the water accommodated fraction (WAF) of either kerosene, gas oil, heavy fuel oil, or crude oil for 96 h. Tissue was collected for RNA extraction and microarray analysis. Exposure to each WAF resulted in a different list of differentially regulated genes, with few genes in common across treatments. Exposure to crude oil WAF changed the expression of genes including cytochrome P4501A (CYP1A) and glutathione-S-transferase (GST) with known roles in detoxification pathways. These gene expression profiles were compared to others from previous experiments that used a diverse suite of toxicants. Clustering algorithms successfully identified gene expression profiles resulting from hydrocarbon exposure. These preliminary analyses highlight the difficulties of using single genes as diagnostic of petroleum hydrocarbon exposures. Further work is needed to determine if multivariate transcriptomic-based biomarkers may be a more effective tool than single gene studies for exposure monitoring of different oils.


What is the phylogenetic position of Cryptosporidium?
The phylogenetic position of Cryptosporidium is elusive. Although previous studies based solely upon small-subunit (SSU) rRNA sequences suggested that the genus was an early emerging lineage among the Apicomplexa, bootstrap support for this placement was low. Here, the phylogenetic position of Cryptosporidium has been re-evaluated for SSU rRNA, fused SSU/large-subunit (LSU) rRNA and six protein sequences using traditional distance-based neighbour-joining, maximum-parsimony and maximum-likelihood methods of phylogenetic reconstruction as well as the new Slow-Fast analysis, which focuses upon the slowly evolving positions within sequences and is especially useful if a long-branch attraction (LBA) artefact is suspected. All the methods of reconstruction indicated a trend for the early emergence of Cryptosporidium at the base of the Apicomplexa and showed that an LBA artefact plays no role in this placement. Although the inclusion of additional numbers of neither species nor genes has significantly enhanced the bootstrap support for this phylogenetic position, recent biochemical, molecular and ultrastructural data are congruent with it. Therefore, we favour a working hypothesis that this genus constitutes an early emerging branch of the Apicomplexa.


Optical coherence tomography reflects brain atrophy in multiple sclerosis: A four-year study.
The aim of this work was to determine whether atrophy of specific retinal layers and brain substructures are associated over time, in order to further validate the utility of optical coherence tomography (OCT) as an indicator of neuronal tissue damage in patients with multiple sclerosis (MS). Cirrus high-definition OCT (including automated macular segmentation) was performed in 107 MS patients biannually (median follow-up: 46 months). Three-Tesla magnetic resonance imaging brain scans (including brain-substructure volumetrics) were performed annually. Individual-specific rates of change in retinal and brain measures (estimated with linear regression) were correlated, adjusting for age, sex, disease duration, and optic neuritis (ON) history. Rates of ganglion cell + inner plexiform layer (GCIP) and whole-brain (r = 0.45; p < 0.001), gray matter (GM; r = 0.37; p < 0.001), white matter (WM; r = 0.28; p = 0.007), and thalamic (r = 0.38; p < 0.001) atrophy were associated. GCIP and whole-brain (as well as GM and WM) atrophy rates were more strongly associated in progressive MS (r = 0.67; p < 0.001) than relapsing-remitting MS (RRMS; r = 0.33; p = 0.007). However, correlation between rates of GCIP and whole-brain (and additionally GM and WM) atrophy in RRMS increased incrementally with step-wise refinement to exclude ON effects; excluding eyes and then patients (to account for a phenotype effect), the correlation increased to 0.45 and 0.60, respectively, consistent with effect modification. In RRMS, lesion accumulation rate was associated with GCIP (r = -0.30; p = 0.02) and inner nuclear layer (r = -0.25; p = 0.04) atrophy rates. Over time GCIP atrophy appears to mirror whole-brain, and particularly GM, atrophy, especially in progressive MS, thereby reflecting underlying disease progression. Our findings support OCT for clinical monitoring and as an outcome in investigative trials.


Cloning and molecular characterization of plasmid-encoded antigens of Borrelia burgdorferi.
Thirteen independent clones that encode Borrelia burgdorferi antigens utilizing antiserum from infection-immune rabbits were identified. The serum was adsorbed against noninfectious B. burgdorferi B31 to enrich for antibodies directed against either infection-associated antigens of B. burgdorferi B31 or proteins preferentially expressed during mammalian infection. The adsorption efficiency of the immune rabbit serum (IRS) was assessed by Western immunoblot analysis with protein lysates derived from infectious and noninfectious B. burgdorferi B31. The adsorbed IRS was used to screen a B. burgdorferi expression library to identify immunoreactive phage clones. Clones were then expressed in Escherichia coli and subsequently analyzed by Western blotting to determine the molecular mass of the recombinant B. burgdorferi antigens. Southern blot analysis of the 13 clones indicated that 10 contained sequences unique to infectious B. burgdorferi. Nucleotide sequence analysis indicated that the 13 clones were composed of 9 distinct genetic loci and that all of the genes identified were plasmid encoded. Five of the clones carried B. burgdorferi genes previously identified, including those encoding decorin binding proteins A and B (dbpAB), a rev homologue present on the 9-kb circular plasmid (cp9), a rev homologue from the 32-kb circular plasmid (cp32-6), erpM, and erpX. Additionally, four previously uncharacterized loci with no known homologues were identified. One of these unique clones encoded a 451-amino-acid lipoprotein with 21 consecutive, invariant 9-amino-acid repeats near the amino terminus that we have designated VraA (for "virulent strain-associated repetitive antigen A"). Since all the antigens identified are recognized by serum from infection immune rabbits, these antigens represent potential vaccine candidates and, based on the identification of dbpAB in this screen, may also be involved in pathogenic processes operative in Lyme borreliosis.


Activation and expression of μ-calpain in dorsal root contributes to RTX-induced mechanical allodynia.
Background Calpain is a calcium-dependent cysteine protease, and inhibition of calpain by pre-treatment with MDL28170 attenuated the rat mechanical allodynia in a variety of pain models. Postherpetic neuralgia (Shingles) is a neuropathic pain conditioned with the presence of profound mechanical allodynia. Systemic injection of resiniferatoxin can reproduce the clinical symptoms of postherpetic neuralgia. In this study, we determined to study whether activation of calpain contributes to cleave the myelin basic protein of dorsal root and is involved in resiniferatoxin-induced mechanical allodynia of postherpetic neuralgia animal model. Results Resiniferatoxin up-regulated the expression and activation of µ-calpain in dorsal root. The expression of µ-calpain was located in Schwann cell of dorsal root, and resiniferatoxin increased the expression of µ-calpain in Schwann cell in L4-L6 dorsal root at six weeks after injection. Resiniferatoxin also induced myelin basic protein degradation in L4-L6 dorsal root at six weeks after injection. Moreover, intraperitoneal injection of calpain inhibitor MDL28170 prevented the degradation of myelin basic protein and then reduced the sprouting of myelinated afferent fibers into spinal lamina II, thus relieving resiniferatoxin-induced mechanical allodynia. Conclusions Up-regulation and activation of µ-calpain located in Schwann cell may be the mechanism underlying resiniferatoxin-mediated proteolysis of myelin basic protein in dorsal root. Calpain inhibitor MDL28170 prevents resiniferatoxin-induced sprouting of myelinated afferent fibers and mechanical allodynia through inhibition of degradation of the myelin basic protein in dorsal root. Our results indicate that inhibition of pathological µ-calpain activation may present an interesting novel drug target in the treatment of postherpetic neuralgia.