Stable aerobic and anaerobic coexistence in anoxic marine zones.
Mechanistic description of the transition from aerobic to anaerobic metabolism is necessary for diagnostic and predictive modeling of fixed nitrogen loss in anoxic marine zones (AMZs). In a metabolic model where diverse oxygen- and nitrogen-cycling microbial metabolisms are described by underlying redox chemical reactions, we predict a transition from strictly aerobic to predominantly anaerobic regimes as the outcome of ecological interactions along an oxygen gradient, obviating the need for prescribed critical oxygen concentrations. Competing aerobic and anaerobic metabolisms can coexist in anoxic conditions whether these metabolisms represent obligate or facultative populations. In the coexistence regime, relative rates of aerobic and anaerobic activity are determined by the ratio of oxygen to electron donor supply. The model simulates key characteristics of AMZs, such as the accumulation of nitrite and the sustainability of anammox at higher oxygen concentrations than denitrification, and articulates how microbial biomass concentrations relate to associated water column transformation rates as a function of redox stoichiometry and energetics. Incorporating the metabolic model into an idealized two-dimensional ocean circulation results in a simulated AMZ, in which a secondary chlorophyll maximum emerges from oxygen-limited grazing, and where vertical mixing and dispersal in the oxycline also contribute to metabolic co-occurrence. The modeling approach is mechanistic yet computationally economical and suitable for global change applications.


Interactions between endogenous and exogenous attention during vigilance.
The ability to remain vigilant over long periods of time is critical for many everyday tasks, but controlled studies of visual sustained attention show that performance declines over time when observers are required to respond to rare stimulus events (targets) occurring in a sequence of standard stimulus events (nontargets). When target discrimination is perceptually difficult, this vigilance decrement manifests as a decline in perceptual sensitivity. We examined whether sudden-onset stimuli could act as exogenous attentional cues to improve sensitivity during a traditional sustained attention task. Sudden-onset cues presented immediately before each stimulus attenuated the sensitivity decrement, but only when stimulus timing (the interstimulus interval [ISI]) was constant. When stimulus timing was variable, exogenous cues increased overall sensitivity but did not prevent performance decline. Finally, independent of the effects of sudden onsets, a constant ISI improved vigilance performance. Our results demonstrate that exogenous attention enhances perceptual sensitivity during vigilance performance, but that this effect is dependent on observers' being able to predict the timing of stimulus events. Such a result indicates a strong interaction between endogenous and exogenous attention during vigilance. We relate our findings to a resource model of vigilance, as well as to theories of endogenous and exogenous attention over short time periods.


Unilateral lower limb hypoplasia in arterial fibromuscular dysplasia.
Hypoplasia or hyperplasia of limbs can be unilateral or bilateral and can involve the upper or lower extremities in any combination. The etiology is varied, and the presentation, clinical features, and investigative pathway differ to some extent among patients. Reported here is a child, referred for limb lengthening, with claudicating unilateral leg pain and unilateral hypoplasia of the same limb. Investigations revealed multiple arterial stenoses consistent with fibromuscular dysplasia. In such cases, early diagnosis and intervention with angioplasty may enhance subsequent growth of the limb, thus avoiding the need for limb lengthening procedures.


Raclopride-induced motor consolidation impairment in primates: role of the dopamine type-2 receptor in movement chunking into integrated sequences.
Results obtained in patients with schizophrenia have shown that antipsychotic drugs may induce motor learning deficits correlated with the striatal type-2 dopamine receptors (D(2)R) occupancy. Other findings suggest that the role of the striatum in motor learning could be related to a process of "chunking" discrete movements into motor sequences. We therefore hypothesized that a D(2)R blocking substance, such as raclopride, would affect motor learning by specifically disrupting the grouping of movements into sequences. Two monkeys were first trained to perform a baseline-overlearned sequence (Seq. A) drug free. Then, a new sequence was learned (Seq. B) and the overlearned sequence was recalled OFF-drug (Seq. A recall OFF-drug). The effect of raclopride was then assessed on the learning of a third sequence (Seq. C), and on the recall of the overlearned sequence (Seq. A recall ON-drug). Results showed that performance related to the overlearned sequence remained the same in the three experimental conditions (Seq. A, Seq. A recall OFF-drug, Seq. A recall ON-drug), whether the primates received raclopride or not. On the other hand, new sequence learning was significantly affected during raclopride treatment (Seq. C), when compared with new sequence learning without the effect of any drug (Seq. B). Raclopride-induced disturbances consisted in performance fluctuations, which persisted even after many days of trials, and prevented the monkeys from reaching a stable level of performance. Further analyses also showed that these fluctuations appeared to be related to monkeys' inability to group movements into single flowing motor sequences. The results of our study suggest that dopamine is involved in the stabilization or consolidation of motor performances, and that this function would involve a chunking of movements into well-integrated sequences.


Isomer-Resolved Imaging of Prostate Cancer Tissues Reveals Specific Lipid Unsaturation Profiles Associated With Lymphocytes and Abnormal Prostate Epithelia.
Prostate cancer is the fourth most common cancer worldwide with definitive diagnosis reliant on biopsy and human-graded histopathology. As with other pathologies, grading based on classical haematoxylin and eosin (H&E) staining of formalin fixed paraffin-embedded material can be prone to variation between pathologists, prompting investigation of biomolecular markers. Comprising around 50% of cellular mass, and with known metabolic variations in cancer, lipids provide a promising target for molecular pathology. Here we apply isomer-resolved lipidomics in combination with imaging mass spectrometry to interrogate tissue sections from radical prostatectomy specimens. Guided by the histopathological assessment of adjacent tissue sections, regions of interest are investigated for molecular signatures associated with lipid metabolism, especially desaturation and elongation pathways. Monitoring one of the most abundant cellular membrane lipids within these tissues, phosphatidylcholine (PC) 34:1, high positive correlation was observed between the n-9 isomer (site of unsaturation 9-carbons from the methyl terminus) and epithelial cells from potential pre-malignant lesions, while the n-7 isomer abundance was observed to correlate with immune cell infiltration and inflammation. The correlation of lipid isomer signatures with human disease states in tissue suggests a future role for isomer-resolved mass spectrometry imaging in assisting pathologists with prostate cancer diagnoses and patient stratification.


Incidence of blood transfusion requirement and factors associated with transfusion following liver lobectomy in dogs and cats: 72 cases (2007-2015).
OBJECTIVE To determine the incidence of blood transfusion, mortality rate, and factors associated with transfusion in dogs and cats undergoing liver lobectomy. DESIGN Retrospective case series. ANIMALS 63 client-owned dogs and 9-client owned cats that underwent liver lobectomy at a specialty veterinary practice from August 2007 through June 2015. PROCEDURES Medical records were reviewed and data extracted regarding dog and cat signalment, hematologic test results before and after surgery, surgical method, number and identity of lobes removed, concurrent surgical procedures, hemoabdomen detected during surgery, incidence of blood transfusion, and survival to hospital discharge (for calculation of mortality rate). Variables were compared between patients that did and did not require transfusion. RESULTS 11 of 63 (17%) dogs and 4 of 9 cats required a blood transfusion. Mortality rate was 8% for dogs and 22% for cats. Pre- and postoperative PCV and plasma total solids concentration were significantly lower and mortality rate significantly higher in dogs requiring transfusion than in dogs not requiring transfusion. Postoperative PCV was significantly lower in cats requiring transfusion than in cats not requiring transfusion. No significant differences in any other variable were identified between dogs and cats requiring versus not requiring transfusion. CONCLUSIONS AND CLINICAL RELEVANCE Dogs and cats undergoing liver lobectomy had a high requirement for blood transfusion, and a higher requirement for transfusion should be anticipated in dogs with perioperative anemia and cats with postoperative anemia. Veterinarians performing liver lobectomies in dogs and cats should have blood products readily available.


One-plasmid double-expression system for preparation of MS2 virus-like particles packaging SARS-CoV-2 RNA.
COVID-19 is a disease caused by a virus named SARS-CoV-2. SARS-CoV-2 is a single-stranded positive-sense RNA virus. Reverse transcription quantitative PCR (RT-qPCR) assays are the gold standard molecular test for detection of RNA viruses. The aim of this study was to construct an RNA-positive control based on MS2 phage-like particles (MS2 VLPs) to detect SARS-CoV-2 RNA. pCDFDuet-1 was used as a one-plasmid double-expression system to construct MS2 VLPs containing ssRNA of SARS-CoV-2. The sequence encoding one copy of maturase, His-tag and coat protein dimer was cloned and inserted into MCS1 of the plasmid; the fragment encoding protein N and ORF1ab from SARS-CoV-2 was cloned and inserted into MCS2. The prepared plasmid was transformed into Escherichia coli strain BL2 (DE3), and expression of the construct was induced by 1 mM isopropyl-L-thio-D-galactopyranoside (IPTG) at 30°C for 12 hours. MS2 VLPs were purified and collected with Ni-NTA affinity chromatography columns. The size and shape of the MS2 VLPs were verified by transmission electron microscopy, and the stability of MS2 VLP packaged RNA was evaluated by treatment with RNase A. Effects of storage temperature and buffer on MS2 VLP stability were also investigated. The results showed that SARS-CoV-2 MS2 VLPs could be successfully produced by this one-plasmid double-expression system. MS2 VLPs showed high stability and may be used as a positive control in molecular diagnosis of COVID-19.


Cost-effectiveness and programmatic benefits of maternal vaccination against pertussis in England.
Maternal pertussis immunisation was introduced during the pertussis resurgence in England in 2012 as a temporary measure to protect infants too young to be vaccinated. The programme was shown to be safe and highly effective. However, continuation of maternal vaccination as a routine programme requires a cost-effectiveness analysis. The estimated prevented disease burden among mothers and their infants was obtained assuming 89% (95% CI: 19%-99%) vaccine efficacy for mothers and 91% (95% CI: 84%-95%) for infants. Future incidence was projected based on the disease rates in 2010-2012, including the four-year cycle of low and high incidence years. Full probabilistic sensitivity analysis was performed for different scenarios. Assuming a vaccine coverage of 60%, there were 1650 prevented hospitalisations in infants (3.5% discounting, the first 10 years), including 55-60 deaths and ∼20,500 cases among mothers, of which around 1800 would be severe. The annual costs of the programme are £7.3 million assuming a price of £10 per dose and £9.4 million assuming £15 per dose. Using discounting of 3.5%, a 200 year time horizon and a price of £10 per dose (+£7.5 administration costs) only 25% of the iterations were below £30,000 per QALY. Using a 35% higher incidence resulted in 88% of the scenarios below this threshold. Assuming that the incidence remains at the level at the height of 2012, then the programme would be highly cost effective, with an ICER of £16,865 (£12,209-£25,976; price of £10 and 3.5%/3.5% discounting). Maternal vaccination is effective in preventing severe illness and deaths in infants but the cost-effectiveness of the programme is highly dependent on future incidence which is necessarily uncertain. However, the duration and magnitude of protection against transmission afforded by the current acellular vaccines is also uncertain as are the associated effects on future herd immunity. The direct protection offered by the maternal dose provides the only certain way of protecting vulnerable infants from birth.


Cathepsin A is expressed in a cell- and region-specific manner in the testis and epididymis and is not regulated by testicular or pituitary factors.
The epithelial cells of the testis are involved in the production, differentiation, and sustenance of sperm, and those of the epididymis play a major role in sperm maturation, protection, and storage. These tissues express various proteins that respond differently to androgens. Cathepsin A is a multifunctional lysosomal carboxypeptidase that also functions as a protective and an activator protein for neuraminidase and beta-galactosidase. In this study, cathepsin A was immunolocalized by light and electron microscopy using a polyclonal affinity-purified antibody on the testis and epididymis of normal, orchidectomized with or without testosterone supplementation, efferent duct-ligated, and hypophysectomized adult rats. In normal rats, cathepsin A expression was noted in lysosomes of Sertoli and Leydig cells but not in germ cells of the testis, as well as nonciliated cells of the efferent ducts. In the epididymis, a cell- and region-specific distribution of cathepsin A was noted. In experimentally treated animals, no changes were noted in the expression of cathepsin A. Immunolabeling of tissues examined at the electron microscopic level revealed that lysosomes were reactive. These data indicate cell- and region-specific expression of cathepsin A in cells of the testis and epididymis and also indicate that cathepsin A expression is not regulated by testicular or pituitary factors.


Clinical patterns among male transsexual candidates with erotic interest in males.
Male-to-female transsexuals who reported an erotic interest in males showed different patterns of sexual activity. Sexual history was used to categorize a transsexual sample into three groups: 44% abstained from sexual activity (Inactive group), 19% were sexually active but avoided using their penis in sexual activity (Avoidant group), and 37% were sexually active and derived pleasure from their penis (Pleasure group). The groups were compared for differences in gender identification, developmental patterns, and personality. Transsexuals in the Avoidant group showed patterns of traits and experiences that generally conformed to characteristics of the nuclear transsexual. They were dissimilar from the other two groups on measures of feminine functioning, heterosexual history, and fetishism. Transsexuals who interact with males in ways that are viewed as more classically homosexual shared more in common with the transsexual group which abstained from sexual activity with males. Both groups displayed more masculinity in development and more evidence of emotional disturbance. The implications of these findings for diagnosis and treatment are discussed.


Comparative pathogenomic analysis reveals a highly tetanus toxin-producing clade of Clostridium tetani isolates in Japan.
C. tetani is a spore-forming, anaerobic bacterium that produces a toxin causing muscle stiffness and paralysis. Tetanus is preventable with the toxoid vaccine, but it remains a significant public health threat in regions with low vaccine coverage. However, there are relatively few isolates and limited genomic information available worldwide. In Japan, about 100 cases are reported each year, but there have been no nationwide surveys of isolates, and no genomic information from Japanese isolates has been published. In our study, we analyzed the genomes of 151 strains from a limited survey of soil in Kumamoto, Japan. Our findings revealed a high degree of genetic diversity, and we also identified a subset of strains that produced significantly more toxin, which provides new insights into the pathogenesis of tetanus. Our findings lay the foundation for future studies to investigate the distribution and evolution of C. tetani in Japan and neighboring countries.


Development of Inflammatory Bowel Disease in HIV Patients: A Danish Cohort Study (1983-2018) With American Validation (1999-2018).
Human immunodeficiency virus (HIV) infection is associated with several immune-mediated disorders. However, the risk of inflammatory bowel disease (IBD) in people living with HIV (PLWH) remains unclear. We aimed to assess the risk of IBD among PLWH using a nationwide, population-based Danish cohort and to validate findings in a large American insurance-based database. Using Danish registries (1983-2018), we identified 8995 PLWH and age- and sex-matched them to 449,750 HIV-negative individuals. Cox regression analysis was undertaken to calculate hazard ratios (HRs) with 95% confidence intervals (CIs) for IBD diagnosis. Results were stratified by sex, age, and year of HIV diagnosis. Using an American insurance-based cohort, Explorys (1999-2018), we assessed the prevalence odds ratio (OR) and 95% CI of IBD diagnosis in PLWH compared with HIV-negative individuals. IBD diagnosis among PLWH in Denmark was increased (HR: 2.25, 95% CI: 1.78-2.83) compared with matched HIV-negative individuals. This was seen for both Crohn's disease (HR: 2.25, 95% CI: 1.47-3.44) and ulcerative colitis (HR: 2.24, 95% CI: 1.70-2.96) and in male (HR: 2.75, 95% CI: 2.15-3.52) but not female (HR: 0.93, 95% CI: 0.48-1.79) PLWH. Explorys analysis also showed an increased odds of IBD diagnoses among PLWH (OR: 1.41; 95% CI: 1.35-1.49). This study finds an increased risk of IBD diagnosis among PLWH in both a Danish and US cohort, highlighting a need to consider IBD in PLWH with new-onset gastrointestinal symptoms. Further research into the role of antiretroviral therapy in this relationship is required.


Oxygen tolerance of an implantable polymer/enzyme composite glutamate biosensor displaying polycation-enhanced substrate sensitivity.
Biosensors were fabricated at neutral pH by sequentially depositing the polycation polyethyleneimine (PEI), the stereoselective enzyme L-glutamate oxidase (GluOx) and the permselective barrier poly-ortho-phenylenediamine (PPD) onto 125-microm diameter Pt wire electrodes (Pt/PEI/GluOx/PPD). These devices were calibrated amperometrically at 0.7 V versus SCE to determine the Michaelis-Menten parameters for enzyme substrate, l-glutamate (Glu) and co-substrate, dioxygen. The presence of PEI produced a 10-fold enhancement in the detection limit for Glu (approximately 20 nM) compared with the corresponding PEI-free configurations (Pt/GluOx/PPD), without undermining their fast response time (approximately 2 s). Most remarkable was the finding that, although some designs of PEI-containing biosensors showed a 10-fold increase in linear region sensitivity to Glu, their oxygen dependence remained low.


A Proteomics-Based Assessment of Inflammation Signatures in Endotoxemia.
We have previously shown that multimers of plasma pentraxin-3 (PTX3) were predictive of survival in patients with sepsis. To characterize the release kinetics and cellular source of plasma protein changes in sepsis, serial samples were obtained from healthy volunteers (n = 10; three time points) injected with low-dose endotoxin (lipopolysaccharide [LPS]) and analyzed using data-independent acquisition MS. The human plasma proteome response was compared with an LPS-induced endotoxemia model in mice. Proteomic analysis of human plasma revealed a rapid neutrophil degranulation signature, followed by a rise in acute phase proteins. Changes in circulating PTX3 correlated with increases in neutrophil-derived proteins following LPS injection. Time course analysis of the plasma proteome in mice showed a time-dependent increase in multimeric PTX3, alongside increases in neutrophil-derived myeloperoxidase (MPO) upon LPS treatment. The mechanisms of oxidation-induced multimerization of PTX3 were explored in two genetic mouse models: MPO global knock-out (KO) mice and LysM Cre Nox2 KO mice, in which NADPH oxidase 2 (Nox2) is only deficient in myeloid cells. Nox2 is the enzyme responsible for the oxidative burst in neutrophils. Increases in plasma multimeric PTX3 were not significantly different between wildtype and MPO or LysM Cre Nox2 KO mice. Thus, PTX3 may already be stored and released in a multimeric form. Through in vivo neutrophil depletion and multiplexed vascular proteomics, PTX3 multimer deposition within the aorta was confirmed to be neutrophil dependent. Proteomic analysis of aortas from LPS-injected mice returned PTX3 as the most upregulated protein, where multimeric PTX3 was deposited as early as 2 h post-LPS along with other neutrophil-derived proteins. In conclusion, the rise in multimeric PTX3 upon LPS injection correlates with neutrophil-related protein changes in plasma and aortas. MPO and myeloid Nox2 are not required for the multimerization of PTX3; instead, neutrophil extravasation is responsible for the LPS-induced deposition of multimeric PTX3 in the aorta.


Expression of superoxide dismutase, catalase and glutathione peroxidase in the bovine corpus luteum: evidence supporting a role for oxidative stress in luteolysis.
Apoptosis, a type of physiological or active cell death, has been implicated as a mechanism underlying regression of the corpus luteum (CL) in the rat, bovine, rabbit and ovine ovary. Previousin vitro studies of cultured luteal cells have also provided evidence which suggests that reactive oxygen species play an important role in luteolysis in the rodent ovary. To further evaluate the potential role of oxidative stress in luteal cell demise, changes in the expression of several enzymes known to protect cells from oxidative stress were investigated using bovine CL collected from ovaries of non-pregnant (day 21 of the estrous cycle; regressed CL) and pregnant (day 21 of pregnancy; functional CL) animals. Biochemical analysis of genomic DNA extracted from these two pools of CL demonstrated the presence of extensive levels of internucleosomal DNA cleavage characteristic of cell death via apoptosis in regressed, but not in functional, CL. Northern blot analysis of total RNA indicated that functional CL expressed significantly higher levels of mRNA encoding secreted superoxide dismutase (SEC-SOD, 1.9 kb) and manganese-containing or mitochondrial SOD (Mn-SOD, multiple transcripts) as compared to regressed CL. Similarly, levels of mRNA encoding catalase (2.1 kb), an enzyme responsible for detoxification of peroxides to water, were significantly higher in functional versus regressed CL. From these data, we conclude that a decline in expression of specific oxidative response genes occurs during luteolysis, and that maintained expression of these genes in the CL during pregnancy may prevent oxidative damage and delay regression.


Genome maintenance in Saccharomyces cerevisiae: the role of SUMO and SUMO-targeted ubiquitin ligases.
The genome of the cell is often exposed to DNA damaging agents and therefore requires an intricate well-regulated DNA damage response (DDR) to overcome its deleterious effects. The DDR needs proper regulation for its timely activation, repression, as well as appropriate choice of repair pathway. Studies in Saccharomyces cerevisiae have advanced our understanding of the DNA damage response, as well as the mechanisms the cell employs to maintain genome stability and how these mechanisms are regulated. Eukaryotic cells utilize post-translational modifications as a means for fine-tuning protein functions. Ubiquitylation and SUMOylation involve the attachment of small protein molecules onto proteins to modulate function or protein-protein interactions. SUMO in particular, was shown to act as a molecular glue when DNA damage occurs, facilitating the assembly of large protein complexes in repair foci. In other instances, SUMOylation alters a protein's biochemical activities, and interactions. SUMO-targeted ubiquitin ligases (STUbLs) are enzymes that target SUMOylated proteins for ubiquitylation and subsequent degradation, providing a function for the SUMO modification in the regulation and disassembly of repair complexes. Here, we discuss the major contributions of SUMO and STUbLs in the regulation of DNA damage repair pathways as well as in the maintenance of critical regions of the genome, namely rDNA regions, telomeres and the 2 μm circle in budding yeast.


Rapid Characterization of Magnetic Moment of Cells for Magnetic Separation.
NCI-H1650 lung cancer cell lines labeled with magnetic nanoparticles via the Epithelial Cell Adhesion Molecule (EpCAM) antigen were previously shown to be captured at high efficiencies by a microfabricated magnetic sifter. If fine control and optimization of the magnetic separation process is to be achieved, it is vital to be able to characterize the labeled cells' magnetic moment rapidly. We have thus adapted a rapid prototyping method to obtain the saturation magnetic moment of these cells. This method utilizes a cross-correlation algorithm to analyze the cells' motion in a simple fluidic channel to obtain their magnetophoretic velocity, and is effective even when the magnetic moments of cells are small. This rapid characterization is proven useful in optimizing our microfabricated magnetic sifter procedures for magnetic cell capture.


Switching to IgG3, IgG2b, and IgA is division linked and independent, revealing a stochastic framework for describing differentiation.
LPS was used to induce switching of B cells to IgG3 and, in the presence of TGF-beta, to IgG2b and IgA. Switching to all three isotypes increased with division number according to a consistent relationship that was independent of time in culture. The mode of activation altered the relationship with division, as CD40 ligand increased switching to IgA and decreased switching to IgG2b and IgG3 when measured per division. This division-linked switching behavior could be described by Gaussian probability distributions centered around a mean division number. The divisions at which switching to IgG3 and IgG2b occurred overlapped, raising the possibility that the two switching mechanisms were linked. However, when IgG3+ and IgG3- B cells were sorted and placed back in culture, they switched to IgG2b at an equivalent rate, indicating that alternative switching decisions were made independently within a single cell. As a consequence, isotype switching could be predicted at the population level by standard probability laws. Therefore, division number provides a framework for a stochastic description of differentiation that may be widely applicable.


A compact system of small planets around a former red-giant star.
Planets that orbit their parent star at less than about one astronomical unit (1 AU is the Earth-Sun distance) are expected to be engulfed when the star becomes a red giant. Previous observations have revealed the existence of post-red-giant host stars with giant planets orbiting as close as 0.116 AU or with brown dwarf companions in tight orbits, showing that these bodies can survive engulfment. What has remained unclear is whether planets can be dragged deeper into the red-giant envelope without being disrupted and whether the evolution of the parent star itself could be affected. Here we report the presence of two nearly Earth-sized bodies orbiting the post-red-giant, hot B subdwarf star KIC 05807616 at distances of 0.0060 and 0.0076 AU, with orbital periods of 5.7625 and 8.2293 hours, respectively. These bodies probably survived deep immersion in the former red-giant envelope. They may be the dense cores of evaporated giant planets that were transported closer to the star during the engulfment and triggered the mass loss necessary for the formation of the hot B subdwarf, which might also explain how some stars of this type did not form in binary systems.


Changes in phytochemical properties and water use efficiency of peppermint (Mentha piperita L.) using superabsorbent polymer under drought stress.
Water consumption management and the application of advanced techniques in the agricultural sector can significantly contribute to the efficient utilization of limited water resources. This can be achieved by improving soil texture, increasing water retention, reducing erosion, and enhancing seedling germination through the use of superabsorbent polymers. This study aimed to investigate the effect of Aquasource superabsorbent (AS) on the morphological characteristics, phytochemical properties, antioxidant content, and water use efficiency of peppermint. It was conducted under different irrigation management and using different superabsorbent levels. Therefore, a 3 × 4 factorial design was used to determine the effects of irrigation intervals (2-, 4-, and 6-day) and different levels of AS amount (zero [control], 0.5, 1, and 2 wt%). The effects of these factors on various parameters (morphological characteristics, essential oil percentage, nutrient, protein, proline, carotenoid, antioxidant, and chlorophyll content, leaf area index, relative water content, and water use efficiency [WUE]) were evaluated. The results showed that morphological characteristics and essential oil percentage decreased significantly under drought stress (increasing the irrigation intervals). However, the addition of 0.5 (wt%) AS improved plant growth conditions. Increasing the amount of superabsorbent used to 1 and 2 (wt%) decreased the measured traits, which indicates the creation of unsuitable conditions for plant growth. AS application improved the growth of the root more than the leaf yield of peppermint. A 0.5 (wt%) addition of AS resulted in root length increases of 3, 13, and 15%, respectively, at irrigation intervals of 2, 4, and 6 days, respectively. Additionally, at 0.5 (wt%) AS, root weight increased by 8, 15, and 16% in 2-, 4-, and 6-day irrigation intervals, respectively. Also, the height of the plant increased by 3, 5, and 17% at 2-, 4-, and 6-day irrigation intervals when 0.5 (wt%) of AS was used compared to the control. As well, essential oil percentage increased by 2.14, 2.06, and 1.63% at 2-, 4-, and 6-day irrigation intervals. The nutrient and protein contents decreased as irrigation intervals and AS usage increased, indicating a similar trend. However, compared with the control, the addition of 0.5 (wt%) of AS resulted in some improvements in nutrients and protein. The highest WUE (3.075 kg m-3) was attained in the 4-day irrigation interval and 1 wt% AS addition. This was followed closely by the 2-day irrigation interval with 1 wt% AS addition at 3.025 kg m-3, and the 4-day irrigation interval with 0.5 wt% AS addition, which reached 2.941 kg m-3. Overall, the use of AS in appropriate amounts (0.5 wt%) can reduce water consumption and enhance essential oil yield and WUE in peppermint cultivation in water-scarce arid and semi-arid regions.


[Digitalized thoracic radiography in cancerology. Present contribution apropos of 200 examinations].
The authors report a double study comparing the conventional chest radiograph and digitalised images. The first study was to visualise normal mediastinal structures (the contour of the posterior mediastinum, the azygo-oesophageal recess, the bronchial sub-segments, intermediate, carina, trachea) and showed a superiority of the digitalised image. The second study was on the recognition of radiological abnormalities in lung cancer which showed no precise superiority of the digitalised image in the mediastinum (bronchial tumour, adenopathies) and a current deficiency at the pleuro-parenchymal level (pulmonary nodules, atelectasis pleural effusion). This double study presents parallel results to those obtained by the anglosaxon authors on different prototypes in whom the technology and possibilities were different. The principles, the advantages and the limits of these prototypes were described and in conclusion a chosen technique for the medium term is proposed by the authors.


Natural and anthropogenic sources of chromium, nickel and cobalt in soils impacted by agricultural and industrial activity (Vojvodina, Serbia).
This study reports the contents and sources of chromium, nickel and cobalt, as well as Al, Ca, Mg, K, Fe and Mn in Vojvodina regions Srem and Central Banat area soil samples. Different methods were applied to identify the sources of the studied elements and to classify the latter as geogenic and/or anthropogenic: modified Tessier sequential extraction, calculation ratio of E/Al, XRPD, correlations and cluster analysis. The sampling methodology was according to the GEMAS project. The results show that increasing content of chromium, cobalt and nickel detected in studied soils can be explained by a distribution pattern and the presence of ultramafic and mafic parent rocks, as well as by significant anthropogenic pollution, mainly originating from the industry at some localities. The statistically significant difference between the content of Cr and Ni in soils of Srem and C. Banat is observed. The content of the studied elements is higher in soils of Srem. The normalization to Al indicating a presence of the anthropogenic sources which my significantly affect the content of Cr and Ni in the soils from Beočin, Ruma and Stara Pazova. There is no shown influence of eventual agrochemicals application on the Cr, Ni and Co content.


DNA adduct formation by allyl glycidyl ether.
Glycidyl ethers are reactive epoxides used as components of a variety of epoxy materials. These compounds are known to cause allergic reactions, but since they are generally also genotoxic it would be of interest to evaluate the risk for induction of such effects. Reaction products of allyl glycidyl ether with nucleic acid components were therefore studied. Adduct standards of expected major products in DNA were prepared and assigned to N-7-guanine, N-1- and N-3-adenine and N-3-cytosine. The adducts were characterized by UV spectroscopy, and the adduct to N-1-adenine also by mass spectrometry and nuclear magnetic resonance spectroscopy. In analogy with the formation of corresponding reaction products of other simple epoxides the N-1-adenine adduct rearranged in a base catalysed reaction to N6 and the N-3-cytosine adduct deaminated to form the corresponding N-3-uracil adduct. For allyl glycidyl ether these further reactions of the N-1-adenine and N-3-cytosine adducts were, however, slower than has been observed for corresponding products of other epoxides, but faster than for methylated and ethylated products. In double-stranded salmon testis DNA treated in vitro with allyl glycidyl ether, the major product was found at N-7-guanine, followed by those at N-1-adenine, N-3-adenine and N-3-cytosine (including N-3-uracil). A minor amount of an N6-adenine adduct was also detected, but only after 48 h of reaction. In single-stranded DNA the yield of the N-1-adenine adduct was increased to about the level of the N-7-guanine adduct. The level of the N-3-cytosine adduct was also considerably higher in single-stranded DNA and was the third largest adduct. The reactivity of N-3-adenine was decreased in single-stranded DNA and since other adducts increased the relative yield of this adduct was very low. The N-7-guanine and N-3-adenine adducts were lost from DNA as a consequence of depurination with half-lives in double-stranded DNA at 37 degrees C and pH 7.4 of 38 and 20 h, respectively. The rates of losses (due to depurination or rearrangement) of initially formed adducts in DNA increased in the order N-1-adenine < N-7-guanine approximately N-3-cytosine < N-3-adenine and were faster in single- than in double-stranded DNA. Taking only the rate of formation and chemical stability into consideration, the adducts with N-1-adenine and N-7-guanine seem to be the most promising candidates for monitoring allyl glycidyl ether exposures in vivo.


Novel type II fatty acid biosynthesis (FAS II) inhibitors as multistage antimalarial agents.
Malaria is a potentially fatal disease caused by Plasmodium parasites and poses a major medical risk in large parts of the world. The development of new, affordable antimalarial drugs is of vital importance as there are increasing reports of resistance to the currently available therapeutics. In addition, most of the current drugs used for chemoprophylaxis merely act on parasites already replicating in the blood. At this point, a patient might already be suffering from the symptoms associated with the disease and could additionally be infectious to an Anopheles mosquito. These insects act as a vector, subsequently spreading the disease to other humans. In order to cure not only malaria but prevent transmission as well, a drug must target both the blood- and pre-erythrocytic liver stages of the parasite. P. falciparum (Pf) enoyl acyl carrier protein (ACP) reductase (ENR) is a key enzyme of plasmodial type II fatty acid biosynthesis (FAS II). It has been shown to be essential for liver-stage development of Plasmodium berghei and is therefore qualified as a target for true causal chemoprophylaxis. Using virtual screening based on two crystal structures of PfENR, we identified a structurally novel class of FAS inhibitors. Subsequent chemical optimization yielded two compounds that are effective against multiple stages of the malaria parasite. These two most promising derivatives were found to inhibit blood-stage parasite growth with IC(50) values of 1.7 and 3.0 μM and lead to a more prominent developmental attenuation of liver-stage parasites than the gold-standard drug, primaquine.


Cocoa-based protein and carbohydrate drink decreases perceived soreness after exhaustive aerobic exercise: a pragmatic preliminary analysis.
The purpose of this pragmatic preliminary analysis was to examine the effectiveness of a cocoa-based protein and carbohydrate prototype drink on skeletal muscle damage and perceived soreness after exhaustive exercise. A repeated-measures experimental design was used. Common biomarkers indicative of skeletal muscle damage included creatine kinase (CK), urinary isoprostanes and inflammatory markers (IL-6, IL-8, C-Reactive Protein [CRP]). Self-reported perception of postexercise soreness was also evaluated. Seven men participated in an exercise session consisting of a 30-minute run on a declined treadmill (-10% grade). Running speed was adjusted accordingly so that participants consistently maintained 75% maximal heart rate. Drinks were ingested immediately after exercise, 2 hours postexercise, and before bed. Blood draws were sampled 30, 60, 120, and 360 minutes postexercise; urine was collected 24 and 48 hours postexercise. A perceived soreness questionnaire was administered 24 and 48 hours postexercise. The test drink had no effect on IL-6, CK, IL-8, CRP, or urinary isoprostanes (p > 0.05). However, the drink decreased the change in perceived soreness from 24 to 48 hours (p = 0.03). Consuming the drink after exercise resulted in a mean change of 2.6 +/- 6 compared to 13.7 +/- 10 for the control. In summary, the drink was effective in decreasing the level of self-reported perceived soreness after exhaustive exercise.


High-dose chemotherapy for children and young adults with stage IV rhabdomyosarcoma.
Rhabdomyosarcoma is the most common soft tissue sarcoma of childhood. Prognosis for patients with metastatic disease has not improved significantly in the past decades. High-dose chemotherapy (HDC) seems to be an attractive option to treat minimal residual disease in metastatic rhabdomyosarcoma patients. The objective of the review was to assess the effectiveness of HDC with stem cell rescue (SRC) versus standard-dose chemotherapy in improving event-free survival (EFS) and overall survival (OS) of children and young adults with metastatic rhabdomyosarcoma. We searched the databases of MEDLINE (1966 to December 2009), EMBASE (1980 to December 2009) and CENTRAL (The Cochrane Library Issue 1, 2009). In addition, we handsearched the reference lists of selected papers and conference proceedings of the SIOP, ASPHO and ASCO meetings (all 2000 to 2009). Randomised controlled trials (RCT), prospective or historical controlled clinical trials (CCT), in which HDC with SCR was compared to conventional chemotherapy and prospective case series (non-controlled clinical trials) including at least 20 naive metastatic rhabdomyosarcoma patients Two review authors independently performed the study selection, quality assessment and data extraction. No RCTs could be identified. We identified one prospective CCT, one retrospective CCT and one non-controlled clinical trial. Another CCT has been published as an abstract. All studies have severe methodological limitations, in particular selection bias could not be excluded. One CCT reported a significantly worse OS compared to oral maintenance therapy, however in a subgroup of high-risk patients no difference could be found. The retrospective CCT reported a similar survival for HDC compared to conventional chemotherapy. The non-controlled clinical trial and the CCT reported as a conference proceeding reported survival outcomes comparable to previous studies. Data on toxicity showed more grade 3-4 toxicity in the HDC group. However, there was no difference in the number of toxic deaths. Overall, the results of this review do not justify the use of HDC with SCR as a standard therapy for children with metastatic rhabdomyosarcoma. However, all reported studies were possibly subject to significant bias, especially selection bias. This might have underestimated the measured effect of HDC. As a result, a clinically important excess of adverse risk patients in the HDC arms may explain the non-beneficial effect of HDC. Only a large prospective RCT will be able to answer the question of whether HDC with SCR adds to survival or not definitively.


Semantic Segmentation of Cell Painted Organelles using DeepLabv3plus Model.
Cell painting based high content fluorescence imaging technique offers deep insight into the functional and biological changes in subcellular structures. However, advanced instrumentation and the limited availability of suitable fluorescent dyes restricts the tool to comprehensively characterize the cell morphology. Therefore, generating fluorescent specific organelle images using transmitted light microscopy provides an alternative solution for clinical applications. In this work, the utility of semantic segmentation deep network for predicting the Endoplasmic Reticulum (ER), cytoplasm and nuclei from a composite image is investigated. To perform this study, a public dataset consisting of 3456 composite images are considered from Broad Bioimage Benchmark collection. The pixel wise labeling is carried out with the generated binary masks for ER, cytoplasm and nuclei. DeepLabv3plus architecture with Atrous Spatial Pyramid Pooling (ASPP) and depth wise separable convolution is used as a learning model to perform semantic segmentation. The accuracy and loss function at different learning rates are analyzed and the segmentation results are validated using Jaccard index, mean Boundary F (BF) score and dice index. The trained model achieved 97.86% accuracy with a loss of 0.07 at the learning rate of 0.01. Mean BF score, dice index and Jaccard index for nuclei, ER and cytoplasm are (0.98, 0.94, 0.88), (0.97, 0.82, 0.7) and (0.95, 0.88, 0.66) respectively. The obtained results indicate that the adopted methodology could delineate the subcellular structures by accurately detecting sharp object boundaries. Therefore, this study could be useful for predicting the cell painted images from transmitted light microscopy without the requirement of fluorescent labeling.


Distinctive temporal profiles of detergent-soluble and -insoluble tau and Aβ species in human Alzheimer's disease.
Alzheimer's disease (AD) pathology relevant proteins tau and beta-amyloid (Aβ) exist as an array of post-translationally modified and conformationally altered species with varying abundance, solubility and toxicity. Insoluble neurofibrillary tau tangles and Aβ plaques are end-stage AD hallmarks, yet may carry less disease significance compared to soluble species. At present, it is unclear how soluble and insoluble tau and Aβ relate to each other as well as to disease progression. Here, detergent soluble and insoluble fractions generated from post-mortem human temporal lobe samples (Brodmann area 21) were probed for tau and Aβ markers in immuno-dot assays. Measures were quantified according to diagnosis (AD cf. Non-AD), neuropathological severity, and correlated with disease progression (Braak stages). All markers were elevated within AD cases cf. non-AD controls (p < 0.05) independent of solubility. However, when considered according to neuropathological severity, phospho-tau (detected via CP13 and AT8 antibodies) was elevated early within the soluble fraction (p < 0.05 intermediate cf. low severity) and emerged only later within the insoluble fraction (p < 0.05 high cf. low severity). In contrast, PHF1 phospho-tau, TOC1 reactive tau oligomers and amyloid markers rose within the two fractions simultaneously. Independent of solubility, cognitive correlations were observed for tau makers and for fibrillary amyloid (OC), however only soluble total Aβ was significantly correlated with intellectual impairment. Following the exclusion of end-stage cases, only soluble total Aβ remained correlated with cognition. The data indicate differential rates of protein aggregation during AD progression and confirm the disease relevance of early emerging soluble Aβ species.


Enhanced arbovirus surveillance with deep sequencing: Identification of novel rhabdoviruses and bunyaviruses in Australian mosquitoes.
Viral metagenomics characterizes known and identifies unknown viruses based on sequence similarities to any previously sequenced viral genomes. A metagenomics approach was used to identify virus sequences in Australian mosquitoes causing cytopathic effects in inoculated mammalian cell cultures. Sequence comparisons revealed strains of Liao Ning virus (Reovirus, Seadornavirus), previously detected only in China, livestock-infecting Stretch Lagoon virus (Reovirus, Orbivirus), two novel dimarhabdoviruses, named Beaumont and North Creek viruses, and two novel orthobunyaviruses, named Murrumbidgee and Salt Ash viruses. The novel virus proteomes diverged by ≥ 50% relative to their closest previously genetically characterized viral relatives. Deep sequencing also generated genomes of Warrego and Wallal viruses, orbiviruses linked to kangaroo blindness, whose genomes had not been fully characterized. This study highlights viral metagenomics in concert with traditional arbovirus surveillance to characterize known and new arboviruses in field-collected mosquitoes. Follow-up epidemiological studies are required to determine whether the novel viruses infect humans.


Using Prior Knowledge on Systems Through PBPK to Gain Further Insight into Routine Clinical Data on Trough Concentrations: The Case of Tacrolimus in Chronic Kidney Disease.
Routine therapeutic drug monitoring (TDM) relies heavily on measuring trough drug concentrations. Trough concentrations are affected not only by drug bioavailability and clearance, but also by various patient and disease factors and the volume of distribution. This often makes interpreting differences in drug exposure from trough data challenging. This study aimed to combine the advantages of top-down analysis of therapeutic drug monitoring data with bottom-up physiologically-based pharmacokinetic (PBPK) modeling to investigate the effect of declining renal function in chronic kidney disease (CKD) on the nonrenal intrinsic metabolic clearance (CLint) of tacrolimus as a case example. Data on biochemistry, demographics, and kidney function, along with 1167 tacrolimus trough concentrations for 40 renal transplant patients, were collected from the Salford Royal Hospital's database. A reduced PBPK model was developed to estimate CLint for each patient. Personalized unbound fractions, blood-to-plasma ratios, and drug affinities for various tissues were used as priors to estimate the apparent volume of distribution. Kidney function based on the estimated glomerular filtration rate (eGFR) was assessed as a covariate for CLint using the stochastic approximation of expectation and maximization method. At baseline, the median (interquartile range) eGFR was 45 (34.5-55.5) mL/min/1.73 m2. A significant but weak correlation was observed between tacrolimus CLint and eGFR (r = 0.2, P < 0.001). The CLint declined gradually (up to 36%) with CKD progression. Tacrolimus CLint did not differ significantly between stable and failing transplant patients. Kidney function deterioration in CKD can affect nonrenal CLint for drugs that undergo extensive hepatic metabolism, such as tacrolimus, with critical implications in clinical practice. This study demonstrates the advantages of combining prior system information (via PBPK) to investigate covariate effects in sparse real-world datasets.


Characterization of isolates of Haemophilus paragallinarum from Argentina.
The biochemical and serological properties of Haemophilus paragallinarum isolates recovered from 11 recent outbreaks of infectious coryza in layer hens and one case of swollen-head syndrome in broilers in Argentina are described. Twenty-four isolates had the typical biochemical properties of H. paragallinarum. All isolates were serotyped according to the Page scheme. Ten of the isolates were serovar A, 11 were serovar B, one was serovar C, and two isolates could not be serotyped. The isolates were also examined using a panel of monoclonal antibodies (MAbs) for Page serovars A (one MAb available) and C (three MAbs available). The serovar B isolates all failed to react with any MAb. The serovar C isolate reacted with all three serovar C MAbs but not with the serovar A MAb. Only six of the 10 serovar A isolates reacted with the serovar A MAb. These results indicate that H. paragallinarum isolates from Argentina are antigenically distinct from those examined in other countries, and it is suggested that coryza vaccines intended for use in Argentina may be more effective if based on local strains.


Pressure induced lung injury in a novel in vitro model of the alveolar interface: protective effect of dexamethasone.
The lungs of infants born with congenital diaphragmatic hernia suffer from immaturity as well as the short and long term consequences of ventilator-induced lung injury, including chronic lung disease. Antenatal and postnatal steroids are among current strategies promoted to treat premature lungs and limit long term morbidity. Although studied in whole-animal models, insight into ventilator-induced injury at the alveolar-capillary interface as well as the benefits of steroids, remains limited. The present study utilizes a multi-fluidic in vitro model of the alveolar-interface to analyze membrane disruption from compressive aerodynamic forces in dexamethasone-treated cultures. Human alveolar epithelial cell lines, H441 and A549, were cultured in a custom-built chamber under constant aerodynamic shear followed by introduction of pressure stimuli with and without dexamethasone (0.1μM). On-chip bioelectrical measurements were noted to track changes to the cellular surface and live-dead assay to ascertain cellular viability. Pressure-exposed alveolar cultures demonstrated a significant drop in TEER that was less prominent with an underlying extracellular-matrix coating. Addition of dexamethasone resulted in increased alveolar layer integrity demonstrated by higher TEER values. Furthermore, dexamethasone-treated cells exhibited faster recovery, and the effects of pressure appeared to be mitigated in both cell types. Using a novel in vitro model of the alveolus, we demonstrate a dose-response relationship between pressure application and loss of alveolar layer integrity. This effect appears to be alleviated by dexamethasone and matrix sub-coating.


The health and quality of life outcomes among youth and young adults with cerebral palsy.
Young NL, Rochon TG, McCormick A, Law M, Wedge JH, Fehlings D. The health and quality of life outcomes among youth and young adults with cerebral palsy. To describe the health and quality of life (QoL) of youth and young adults who have cerebral palsy (CP), and to assess the impact of 3 key factors (severity, age, and sex) on these outcomes. Cross-sectional survey. Participants were identified from 6 children's treatment centers in Ontario. The sample of participants (N=199) included youth (n=129; age, 13-17y) and adults (n=70; age, 23-33y) with a broad range of severity: 35% mild, 19% moderate, and 47% severe. Not applicable. Health Utilities Index (HUI(3)), Assessment of Quality of Life (AQoL), and Self-Rated Health (SRH). SRH was reported to be excellent or very good by 57% of youth and 46% of adults. Mean HUI(3) scores were .30 for youth and .31 for adults. Mean AQoL scores were .28 for youth and adults. Severity of CP in childhood predicted 55% of the variance in HUI(3) scores and 45% of the variance in AQoL scores. Age and sex were not significant predictors of health or QoL. The observed health and QoL scores were much lower than those previously reported in the literature. This is likely a result of the inclusion of those with severe CP. The scores for youth were similar to those for adults and suggest that health and QoL outcomes were relatively stable across the transition to adulthood. Youth and adults with CP have limited health status and will require health care support throughout their lives to help them optimize their well being. Longitudinal follow-up studies are essential to understand better the patterns of health in this population over time.


Isolation and mapping the chicken zona pellucida genes: an insight into the evolution of orthologous genes in different species.
The avian oocyte is surrounded by a specialized extracellular glycoproteinaceous matrix, the perivitelline membrane, which is equivalent to the zona pellucida (ZP) in mammals and the chorion in teleosts. A number of related ZP genes encode the proteins that make up this matrix. These proteins play an important role in the sperm/egg interaction and may be involved in speciation. The human genome is known to contain ZP1, ZP2, ZP3, and ZPB genes, while a ZPAX gene has also been identified in Xenopus. The rapid evolution of these genes has confused the nomenclature and thus orthologous relationships across species. In order to clarify these homologies, we have identified ZP1, ZP2, ZPC, ZPB, and ZPAX genes in the chicken and mapped them to chromosomes 5, 14, 10, 6, and 3, respectively, establishing conserved synteny with human and mouse. The amino acid sequences of these genes were compared to the orthologous genes in human, mouse, and Xenopus, and have given us an insight into the evolution of these genes in a variety of different species. The presence of the ZPAX gene in the chicken has highlighted a pattern of probable gene loss by deletion in mouse and gene inactivation by deletion, and base substitution in human.


Using Mobile EEG to Investigate Alpha and Beta Asymmetries During Hand and Foot Use.
The Edinburgh Handedness Inventory (EHI) and the Waterloo Footedness Questionnaire (WFQ) are two of the most widely used questionnaires to assess lateralized everyday behavior in human participants. However, it is unclear to what extent the specific behavior assessed in these questionnaires elicit lateralized neural activity when performed in real-life situations. To illuminate this unresolved issue, we assessed EEG alpha and beta asymmetries during real-life performance of the behaviors assessed in the EHI and WFQ using a mobile EEG system. This methodology provides high ecological validity for studying neural correlates of motor behavior under more naturalistic conditions. Our results indicate that behavioral performance of items of both the EHI and WFQ differentiate between left- and right-handers and left- and right-footers on the neural level, especially in the alpha frequency band. These results were unaffected by movement parameters. Furthermore, we could demonstrate that neural activity elicited specifically during left-sided task performance provides predictive power for the EHI or WFQ score of the participants. Overall, our results show that these prominent questionnaires not only distinguish between different motor preferences on the behavioral level, but also on the neurophysiological level. Furthermore, we could show that mobile EEG systems are a powerful tool to investigate motor asymmetries in ecologically valid situations outside of the laboratory setting. Future research should focus on other lateralized behavioral phenotypes in real-life settings to provide more insights into lateralized motor functions.


Metastatic gastric cancer from malignant fibrous histiocytoma: report of a case.
Metastatic gastric cancer originating from malignant fibrous histiocytoma (MFH) is rare. To our knowledge, only nine other cases have been reported. We report the case of a 75-year-old man who underwent a distal gastrectomy for advanced gastric carcinoma, 2 years after resection of an MFH from the left side of his back. We based our preliminary diagnosis of primary advanced gastric carcinoma on the results of a preoperative biopsy specimen, which suggested either poorly differentiated adenocarcinoma or nonepithelial cell-originating malignant disease-like lymphoma. The resected stomach contained a large ulcerative tumor in the antral section, which was positive for Kp-1 and S-100 by immunohistochemical staining, confirming a pathological diagnosis of metastatic MFH of the stomach. He died of recurrence in the mediastinal space 16 months after the gastrectomy. Our analysis of this and previous cases suggests that resection may be inappropriate for patients with gastric metastasis of MFH because of the extremely high malignant potential of this tumor.


Clinically diagnosed childhood asthma and follow-up of symptoms in a Swedish case control study.
Childhood asthma has risen dramatically not only in the western societies and now forms a major and still increasing public health problem. The aims of this study were to follow up at the age of ten the patterns of asthma symptoms and associations among children with a clinically diagnosed asthma in a sizeable urban-rural community and to in compare them with demographic controls using a standardised questionnaire. In a defined region in Sweden with a population of about 150,000 inhabitants, all children (n = 2,104) born in 1990 were recorded. At the age of seven all primary care and hospital records of the 1,752 children still living in the community were examined, and a group of children (n = 191) was defined with a well-documented and medically confirmed asthma diagnosis. At the age of ten, 86 % of these cases (n = 158) and controls (n = 171) completed an ISAAC questionnaire concerning asthma history, symptoms and related conditions. Different types of asthma symptoms were highly and significantly over-represented in the cases. Reported asthma heredity was significantly higher among the cases. No significant difference in reported allergic rhinitis or eczema as a child was found between cases and controls. No significant difference concerning social factors or environmental exposure was found between case and controls. Among the control group 4.7 % of the parents reported that their child actually had asthma. These are likely to be new asthma cases between the age of seven and ten and give an estimated asthma prevalence rate at the age of ten of 15.1 % in the studied cohort. A combination of medical verified asthma diagnosis through medical records and the use of self-reported symptom through the ISAAC questionnaire seem to be valid and reliable measures to follow-up childhood asthma in the local community. The asthma prevalence at the age of ten in the studied birth cohort is considerably higher than previous reports for Sweden. Both the high prevalence figure and allowing the three-year lag phase for further settling of events in the community point at the complementary roles of both hospital and primary care in the comprehensive coverage and control of childhood asthma in the community.


New Delhi metallo-β-lactamase and extended spectrum β-lactamases co-producing isolates are high in community-acquired urinary infections in Assam as detected by a novel multiplex polymerase chain reaction assay.
The ability of microorganisms to evade antibiotic pressure is challenging in healthcare as patients have little or no drug treatment options. Detection of the prevalence of antibacterial resistance pattern helps towards improved antibiotic policy and empirical treatment. We carried out antibiogram profiling and documented the prevalence and co-prevalence of New Delhi metallo-β-lactamase (NDM) and extended spectrum β-lactamases (ESBL) encoding genes in urinary Escherichia coli and Klebsiella pneumonia isolates. Antibiotic susceptibilities were tested for 241 isolates of E. coli and K. pneumoniae from urine samples collected from out- and hospitalised patients. Polymerase chain reaction (PCR) was carried out on isolates tested positive for phenotypic production of metallo-β-lactamase and ESBL. A multiplex PCR assay was designed to detect the genes. Multiplex PCR assay designed had a limit of detection of 10 3 CFU/mL in vitro. NDM detected was significantly higher among K. pneumoniae compared to E. coli (69.2% vs. 18.2%; P = 0.001). Of 17, 14 NDM positive isolates also harboured ESBL genes. The co-production of CTX-M + TEM + NDM (3/9; 33.3% and 5/8; 62.5%) was most common in K. pneumoniae and E. coli, respectively while CTX-M + TEM + SHV + NDM was found in one isolate. Of the 156 phenotypically ESBL producing isolates, CTX-M, TEM and SHV was detected by PCR in 85, 53 and 24 isolates, respectively. NDM and ESBL co-producing isolates were both community (64.7%) and hospital (35.29%) acquired among E. coli. Antibiotic resistance can be effectively evaluated by a cost and time effective molecular method, such as the multiplex PCR used in this study, which complement culture and sensitivity tests.


Suicidal ideation and perceived burdensomeness in patients with chronic pain.
There is a clear relationship between suicide risk and chronic pain conditions. However, the exact nature of this link has been poorly understood, with risk attribution often limited to comorbid depression. Perceived burdensomeness has already been confirmed as a risk factor for suicidal ideation (SI) and suicide attempt in the general population. Self-perceived burden, studied among medically and terminally ill medical populations, has begun to receive a great deal of attention as a suicide risk factor. However, this risk has not been considered in an outpatient chronic pain population, a group likely to experience perceived burdensomeness as a particular problem. Guidelines recommend routine suicide risk screening in medical settings, but many questionnaires are time-consuming and do not allow for the assessment of the presence of newly identified risk constructs, such as perceived burdensomeness. This retrospective study examined the relationship between depression, perceived burdensomeness, and SI in a patient sample seeking behavioral treatment for chronic pain management. A logistic regression model was developed, with preliminary results indicating perceived burdensomeness was the sole predictor of SI, even in the presence of other well-established risk factors such as age, gender, depressive symptoms, and pain severity. Findings highlight the potential utility of a single-item screening question in routine clinical care as an incrementally superior predictor of SI in a chronic pain population.


Benzylimidazolines as h5-HT1B/1D serotonin receptor ligands: a structure-affinity investigation.
Benzylimidazolines may represent a class of 5-HT1D ligands that has yet to be exploited. On the basis of a previous report that the 2-(substituted-benzyl)imidazoline alpha-adrenergic agonist oxymetazoline (8) binds with high affinity at calf brain 5-HT1D receptors, we explored the structure-affinity relationships of a series of related derivatives. Each of the aromatic substituents was removed and then reinstated in a systematic manner to determine the influence of the individual substituents on binding. It was found that all of the aromatic substituents of 8 act in concert to impart high affinity. However, although the 3-hydroxy group could be removed without significantly reducing affinity for h5-HT1D (i.e., human 5-HT1Dalpha) receptors, this modification reduced h5-HT1B (i.e., human 5-HT1Dbeta) receptor affinity by nearly 50-fold. The 2, 6-dimethyl groups also contribute to binding but seem to play a greater role for h5-HT1B binding than h5-HT1D binding. With the appropriate structural modifications, several compounds were identified that display 20- to >100-fold selectivity for h5-HT1D versus h5-HT1B receptors. Preliminary functional data suggest that these compounds behave as agonists. Given that 5-HT1D agonists are currently being explored for their antimigraine action and that activation of h5-HT1B receptors might be associated with cardiovascular side effects, h5-HT1D-selective agents may offer a new lead for the development of therapeutically efficacious agents.


Differences between Well-Differentiated Neuroendocrine Tumors and Ductal Adenocarcinomas of the Pancreas Assessed by Multi-Omics Profiling.
Most pancreatic neuroendocrine tumors (PNETs) are indolent, while pancreatic ductal adenocarcinomas (PDACs) are particularly aggressive. To elucidate the basis for this difference and to establish the biomarkers, by using the deep sequencing, we analyzed somatic variants across coding regions of 409 cancer genes and measured mRNA/miRNA expression in nine PNETs, eight PDACs, and four intestinal neuroendocrine tumors (INETs). There were 153 unique somatic variants considered pathogenic or likely pathogenic, found in 50, 57, and 24 genes in PDACs, PNETs, and INETs, respectively. Ten and 11 genes contained a pathogenic mutation in at least one sample of all tumor types and in PDACs and PNETs, respectively, while 28, 34, and 11 genes were found to be mutated exclusively in PDACs, PNETs, and INETs, respectively. The mRNA and miRNA transcriptomes of PDACs and NETs were distinct: from 54 to 1659 differentially expressed mRNAs and from 117 to 250 differentially expressed miRNAs exhibited high discrimination ability and resulted in models with an area under the receiver operating characteristics curve (AUC-ROC) >0.9 for both miRNA and mRNA. Given the miRNAs high stability, we proposed exploring that class of RNA as new pancreatic tumor biomarkers.


Are health systems redesigning how health care is delivered?
To explore why and how health systems are engaging in care delivery redesign (CDR)-defined as the variety of tools and organizational change processes health systems use to pursue the Triple Aim. A purposive sample of 24 health systems across 4 states as part of the Agency for Healthcare Research and Quality's Comparative Health System Performance Initiative. An exploratory qualitative study design to gain an "on the ground" understanding of health systems' motivations for, and approaches to, CDR, with the goals of identifying key dimensions of CDR, and gauging the depth of change that is possible based on the particular approaches to redesign care being adopted by the health systems. Semi-structured telephone interviews with health system executives and physician organization leaders from 24 health systems (n = 162). We identify and define 13 CDR activities and find that the health systems' efforts are varied in terms of both the combination of activities they are engaging in and the depth of innovation within each activity. Health system executives who report strong internal motivation for their CDR efforts describe more confidence in their approach to CDR than those who report strong external motivation. Health system leaders face uncertainty when implementing CDR due to a limited evidence base and because of the slower than expected pace of payment change. The ability to validly and reliably measure CDR activities-particularly across varying organizational contexts and markets-is currently limited but is key to better understanding CDR's impact on intended outcomes, which is important for guiding both health system decision making and policy making.


Reliability of immunohistochemical demonstration of oestrogen receptors in routine practice: interlaboratory variance in the sensitivity of detection and evaluation of scoring systems.
To investigate interlaboratory variance in the immunohistochemical (IHC) detection of oestrogen receptors so as to determine the rate of false negatives, which could adversely influence the decision to give adjuvant tamoxifen treatment. To ensure that similar results are obtained by different institutions, 200 laboratories from 26 countries have joined the UK national external quality assessment scheme for immunocytochemistry (NEQAS-ICC). Histological sections from breast cancers having low, medium, and high levels of oestrogen receptor expression were sent to each of the laboratories for immunohistochemical staining. The results obtained were evaluated for the sensitivity of detection, first by estimating threshold values of 1% and 10% of stained tumour cells, and second by the Quick score method, by a panel of four assessors judging individual sections independently on a single blind basis. The results were also evaluated using participants' own threshold values. Over 80% of laboratories were able to demonstrate oestrogen receptor positivity on the medium and high expressing tumours, but only 37% of laboratories scored adequately on the low expressing tumour. Approximately one third of laboratories failed to register any positive staining in this tumour, while one third showed only minimal positivity. There is considerable interlaboratory variability, especially in relation to the detection of breast cancers with low oestrogen receptor positivity, with a false negative rate of between 30% and 60%. This variability appears to be caused by minor differences in methodology that may be rectified by fine adjustment of overall technique.


Risk factors associated with extended-spectrum β-lactamase-producing Enterobacteriaceae nosocomial bloodstream infections in a tertiary care hospital: a clinical and molecular analysis.
To describe the risk factors and molecular epidemiology of nosocomial bloodstream infections caused by extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae in a tertiary care hospital. Patients with enterobacteria-positive blood cultures were included. ESBL expression in the isolates was detected using the combination disk method. Antimicrobial susceptibility testing was performed using the disk diffusion method. bla(SHV), bla(TEM), and bla(CTX-M) genes were identified in the isolated strains by PCR and sequencing. Klebsiella pneumoniae isolates were genotyped by PFGE. Of the 90 isolates recovered, half were found to express ESBLs. Twenty-eight (62%) of these isolates were K. pneumoniae, 8 (18%) were Escherichia coli, 6 (13%) were Enterobacter cloacae, and 3 (7%) were Serratia marcescens. Multivariate logistic regression analysis showed that the only independent risk factor associated with infection by ESBL-producing strains was use of broad-spectrum cephalosporins. None of the isolates was resistant to imipenem. The bla(SHV5) gene was detected in 84% of isolates, followed by bla(CTX-M15) (27%), bla(SHV2) (9%), and bla(SHV12) (7%). PFGE identified six clones among the 28 ESBL-producing K. pneumoniae isolates. ESBL-producing K. pneumoniae clones were detected throughout the hospital. Use of broad-spectrum cephalosporins is the most important risk factor associated with the proliferation of ESBL-producing strains.


Targeted p16(Ink4a) epimutation causes tumorigenesis and reduces survival in mice.
Cancer has long been viewed as a genetic disease; however, epigenetic silencing as the result of aberrant promoter DNA methylation is frequently associated with cancer development, suggesting an epigenetic component to the disease. Nonetheless, it has remained unclear whether an epimutation (an aberrant change in epigenetic regulation) can induce tumorigenesis. Here, we exploited a functionally validated cis-acting regulatory element and devised a strategy to induce developmentally regulated genomic targeting of DNA methylation. We used this system to target DNA methylation within the p16(Ink4a) promoter in mice in vivo. Engineered p16(Ink4a) promoter hypermethylation led to transcriptional suppression in somatic tissues during aging and increased the incidence of spontaneous cancers in these mice. Further, mice carrying a germline p16(Ink4a) mutation in one allele and a somatic epimutation in the other had accelerated tumor onset and substantially shortened tumor-free survival. Taken together, these results provide direct functional evidence that p16(Ink4a) epimutation drives tumor formation and malignant progression and validate a targeted methylation approach to epigenetic engineering.


Polyphenol contents and antioxidant activities of five Indigofera species (Fabaceae) from Burkina Faso.
Aqueous acetone extracts prepared from five Indigofera species of Burkina Faso, namely Indigofera colutea (Burm.) Murril., I. macrocalyx Guilld et Perr., I. nigritana Hook f., I. pulchra willd. and I. tinctoria L., were investigated for their phytochemical composition and their antioxidant activities. Standard methods and TLC were used to screen the phytochemical composition. The total phenolic and flavonoid content of extracts were assessed by Folin-Ciocalteu and AlCl3 methods, respectively. These extracts were also evaluated for their antioxidant potentials using ferric reducing antioxidant power (FRAP), 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulphonate) (ABTS) assays. Flavonoids, saponins, quinones, sterols/triterpenes and tannins were present in all these species except for I. pulchra where quinones were not found. Gallic acid, caffeic acid, rutin and myricetin in I. colutea; gallic acid, quercitrin, myricetin in I. tinctoria; galangin and myricetin in I. macrocalyx were identified by thin layer chromatography. Among these, I. colutea, I. tinctoria, I. nigritana and I. macrocalyx, which had the highest phenolic content, were also found to possess the best antioxidant activities. The results indicated a good correlation between antioxidant activities and total phenolic content (p<0.05 for FRAP/DPPH and DPPH/ABTS and p<0.01 for FRAP/ABTS). These plants represent promising sources of natural antioxidants and these findings give scientific bases to their ethnopharmacological uses.


The psychiatric inpatient physical health assessment sheet (PIPHAS): a useful tool to improve the speed, efficiency, and documentation of physical examination in new psychiatric inpatients.
There is increased morbidity and mortality among patients suffering from mental illness. This is believed to be multi-factorial. Poor access to healthcare, the stigma of mental illness, reduced clinic attendance, lifestyle factors, and side effects of medications are cited as possible contributing factors. It is therefore vital to perform a physical examination to identify previously undiagnosed conditions during the admission of a psychiatric inpatient. The Royal College of Psychiatrists recommends that all patients admitted to a psychiatric hospital should receive a full physical examination on admission, or within twenty-four hours of admission. A snapshot audit was carried out at Prospect Park Hospital in Reading, which highlighted that The Royal College of Psychiatrist's recommendation, along with Trust guidelines regarding physical examination were not being met, with only 78 out of 111 patients (70.3%) undergoing an examination during their admission. In addition to this, examinations were often poorly documented and not covering all examination domains. A psychiatric inpatient physical health assessment sheet (PIPHAS) was designed and introduced, providing a quick and standardised approach to the documentation of a physical examination. After the intervention was put into practice, its impact was assessed by performing a retrospective review of the admission clerking notes of the next 100 admissions to Prospect Park Hospital. Following the introduction of the PIPHAS form there was an increase in the number of patients undergoing physical examination on admission to hospital (75 out of 100 patients, 75%). There was also an increase in the thorough documentation of all examination domains (e.g. respiratory examination) for patients that had a completed PIPHAS form scanned within their medical records. This quality improvement project demonstrates that the PIPHAS form is a useful tool to improve the speed, efficiency, and documentation of a thorough physical examination in new psychiatric inpatients.


PPARbeta/delta regulates the human SIRT1 gene transcription via Sp1.
NAD-dependent deacetylase SIRT1 is known to be activated by caloric restriction and is related to longevity. A natural polyphenolic compound resveratrol is also shown to increases SIRT1 activity and extends lifespan. However, the transcriptional regulation of SIRT1 gene has not completely examined in the context of metabolism. Thus, in this study, we characterized the 5' -flanking region of human SIRT1 gene. We first found that representative metabolic hormones and related factors (glucocorticoid, glucagon/cAMP, and insulin) did not show significant effect on SIRT1 gene transcription. PPARalpha and PPARgamma1 without/with their specific ligands did not have significant effect as well. In contrast, expression of PPARbeta/delta (PPARdelta markedly increased the 5' -promoter activity of SIRT1 gene, which was further amplified by the addition of GW501516, a selective PPARdelta agonist. Deletion/mutation mapping analyses failed to identify PPAR binding element but revealed the presence of canonical Sp1 binding site, which was conserved among species. The Sp1 site is functional, because Sp1 overexpresson significantly enhanced SIRT1 promoter activity, and the binding of Sp1 to the element was confirmed by EMSA and ChIP assays. Interestingly, specific Sp1 antagonist mithramycin completely abolished the PPARdelta-mediated induction of SIRT1 gene transcription. Altogether, our data suggest the predominant role of PPARdelta in the transcriptional regulation of SIRT1 gene. Furthermore, the effects of PPARdelta seem to be mediated by Sp1. We assume that, in vivo, starvation increases lipolysis-derived free fatty acid and activates PPARdelta and the resultant increase in SIRT1 expression, in addition to the activation by NAD and AMPK, facilitates the deacetylation of a variety of proteins involved in mitochondrial beta-oxidation pathway and cell survival.


Breast cancer in Mexican women: an epidemiological study with cervical cancer control.
In Mexico, breast cancer (BC) is one of the main causes of cancer deaths in women, with increasing incidence and mortality in recent years. Therefore, the aim of the study is identify possible risk factors related to BC. An epidemiological study of hospital cases of BC and controls with cervical uterine cancer (CUCA) was carried out at eight third level concentration hospitals in Mexico City. The total of 353 incident cases of BC and 630 controls with CUCA were identified among women younger than 75 years who had been residents of the metropolitan area of Mexico City for at least one year. Diagnosis was confirmed histologically in both groups. Variables were analyzed according to biological and statistical plausibility criteria. Univariate, bivariate and multivariate analyses were carried out. Cases and controls were stratified according to the menopausal hormonal status (pre and post menopause). The factors associated with BC were: higher socioeconomic level (OR= 2.77; 95%CI = 1.77 - 4.35); early menarche (OR= 1.32; 95%CI= 0.88 - 2.00); old age at first pregnancy (>31 years: OR= 5.49; 95%CI= 2.16 - 13.98) and a family history of BC (OR= 4.76; 95% CI= 2.10 - 10.79). In contrast, an increase in the duration of the breastfeeding period was a protective factor (>25 months: OR= 0.38; 95%CI= 0.20 - 0.70). This study contributes to the identification of risk factors for BC described in the international literature, in the population of Mexican women. Breastfeeding appears to play an important role in protecting women from BC. Because of changes in women's lifestyles, lactation is decreasing in Mexico, and young women tend not to breastfeed or to shorten the duration of lactation.


Alzheimer's beta-amyloid peptides compete for insulin binding to the insulin receptor.
The amyloid-beta (Abeta) peptide is neurotoxic and associated with the pathology of Alzheimer's disease (AD). We investigated the effect of Abeta peptides on insulin binding to the insulin receptor because it is known that (1) Abeta and insulin are both amyloidogenic peptides sharing a common sequence recognition motif, (2) Abeta and insulin are substrates for the same insulin degrading enzyme, and (3) impaired glucose metabolism is a characteristic event in the pathology of AD. We discovered that Abeta(1-40) and Abeta(1-42,) the main physiological forms, reduced insulin binding and receptor autophosphorylation. The reduction in binding was caused by a decrease in the affinity of insulin binding to the insulin receptor. This reduction was independent of the receptor concentration. The reverse, control peptide Abeta(40-1) did not reduce insulin binding or insulin receptor autophosphorylation. These results demonstrate that Abeta is a direct competitive inhibitor of insulin binding and action. We speculate that the increased levels of Abeta in Alzheimer's disease may be linked to the associated insulin resistance that has been observed previously in this disease.


Structural and kinetic evidence of aging after organophosphate inhibition of human Cathepsin A.
Human Cathepsin A (CatA) is a lysosomal serine carboxypeptidase of the renin-angiotensin system (RAS) and is structurally similar to acetylcholinesterase (AChE). CatA can remove the C-terminal amino acids of endothelin I, angiotensin I, Substance P, oxytocin, and bradykinin, and can deamidate neurokinin A. Proteomic studies identified CatA and its homologue, SCPEP1, as potential targets of organophosphates (OP). CatA could be stably inhibited by low µM to high nM concentrations of racemic sarin (GB), soman (GD), cyclosarin (GF), VX, and VR within minutes to hours at pH 7. Cyclosarin was the most potent with a kinetically measured dissociation constant (KI) of 2 µM followed by VR (KI = 2.8 µM). Bimolecular rate constants for inhibition by cyclosarin and VR were 1.3 × 103 M-1sec-1 and 1.2 × 103 M-1sec-1, respectively, and were approximately 3-orders of magnitude lower than those of human AChE indicating slower reactivity. Notably, both AChE and CatA bound diisopropylfluorophosphate (DFP) comparably and had KIDFP = 13 µM and 11 µM, respectively. At low pH, greater than 85% of the enzyme spontaneously reactivated after OP inhibition, conditions under which OP-adducts of cholinesterases irreversibly age. At pH 6.5 CatA remained stably inhibited by GB and GF and <10% of the enzyme spontaneously reactivated after 200 h. A crystal structure of DFP-inhibited CatA was determined and contained an aged adduct. Similar to AChE, CatA appears to have a "backdoor" for product release. CatA has not been shown previously to age. These results may have implications for: OP-associated inflammation; cardiovascular effects; and the dysregulation of RAS enzymes by OP.


Disaggregating Hispanic American Cancer Mortality Burden by Detailed Ethnicity.
Hispanics are the largest minority population in the United States (18%). They represent a heterogeneous and growing population. Cancer is the leading cause of death among Hispanics, yet few studies have described cancer mortality burden by specific Hispanic group nationwide. Cancer-related deaths from U.S. death certificates for the years 2003-2012 were analyzed for decedents identifying as Mexican, Puerto Rican, Cuban, and Central or South American. We calculated descriptive statistics, including potential years of lives lost (PYLL), age-adjusted rates, standardized mortality ratios, and fitted JoinPoint regression models, to evaluate annual trends by Hispanic group, using non-Hispanic Whites (NHW) as the reference population. We identified 287,218 cancer-related deaths among Hispanics and 4,570,559 among NHWs. Mortality trends were heterogeneous across Hispanic groups. Female NHWs and male Puerto Ricans had the greatest rates of adjusted PYLL per 1,000 (NHWs, 19.6; Puerto Ricans, 16.5). Liver cancer was ranked among the top 5 cancer-related deaths for every Hispanic group, but not for NHWs. Stomach cancer mortality was twice as high for most Hispanic groups when compared with NHWs and especially high for Mexicans [male standardized mortality ratio (SMR), 2.07; 95% confidence interval (CI), 2.01-2.13; female SMR, 2.62; 95% CI, 2.53-2.71]. We observed marked heterogeneity in cancer mortality across Hispanic groups. Several cancers affect Hispanics disproportionately compared with NHWs. Screening programs in Hispanics should be considered for stomach and liver cancer. Disaggregated analysis of Hispanics is needed to fully understand cancer burden among the diverse Hispanic population and is critical for cancer prevention and control efforts.


Spherically Symmetric Scalar Hair for Charged Black Holes.
The no-hair theorem by Mayo and Bekenstein states that there exists no nonextremal static and spherical charged black hole endowed with hair in the form of a charged scalar field with a self-interaction potential. In our recent work [Phys. Lett. B 803, 135324 (2020)PYLBAJ0370-2693], we showed that the effect of a scalar mass term is important at an asymptotic infinity, which was omitted to prove the no-hair theorem. In this Letter, we demonstrate that there actually exists static and spherical charged scalar hair, dubbed as Q hair, around charged black holes, by taking into account the backreaction to the metric and gauge field. We also discuss that Q cloud, which is constructed without the backreaction around a Reissner-Nordström black hole, is a good approximation to Q hair under a certain limit.


Default and non-compliance among adult epileptics in Zaria, Nigeria. The need to restructure continued care.
Forty five epileptics were observed for non-compliance or default over 2 1/2 years. There were 39 episodes of non-compliance among 19 patients. Six patients had 3 or 4 episodes of non-compliance. Thirteen patients gave their reasons as non-availability of drugs or excessive cost of drugs. Drowsiness or confusion over drug dosages was the reason given by 3 others. Three other patients unilaterally suspended medication. Eighteen patients had defaulted. Twelve of them were males. Three had defaulted after only a single clinic attendance. Seizures in 9 patients were in temporary control immediately prior to default. It is suggested that otherwise well epileptics should be managed in their local community. As a matter of urgency governments need to provide drugs at subsidised rates and provide a mechanism for distributing them efficiently.


Influenza surveillance in animals: what is our capacity to detect emerging influenza viruses with zoonotic potential?
A survey of national animal influenza surveillance programmes was conducted to assess the current capacity to detect influenza viruses with zoonotic potential in animals (i.e. those influenza viruses that can be naturally transmitted between animals and humans) at regional and global levels. Information on 587 animal influenza surveillance system components was collected for 99 countries from Chief Veterinary Officers (CVOs) (n = 94) and published literature. Less than 1% (n = 4) of these components were specifically aimed at detecting influenza viruses with pandemic potential in animals (i.e. those influenza viruses that are capable of causing epidemic spread in human populations over large geographical regions or worldwide), which would have zoonotic potential as a prerequisite. Those countries that sought to detect influenza viruses with pandemic potential searched for such viruses exclusively in domestic pigs. This work shows the global need for increasing surveillance that targets potentially zoonotic influenza viruses in relevant animal species.


[Effect of L-carnitine on metabolic disorders in rats with experimental acyl-CoA dehydrogenase deficiency].
Effect of L-carnitine (LC) on the metabolism of organic acids and carnitine homeostasis was studied in rats with riboflavin deficiency producing unusual dicarboxylic acidurea and modeling multiple acyl-CoA dehydrogenase deficiency in humans. Riboflavin deficient (RFD) rats exhibited increased excretion of glutaric, ethylmalonic, and methylsuccinic acids, as well as isovaleryl-, butyryl-, isobutyryl-, 2-methyl-butyryl-, and hexanoylglycine, short-chain and medium-chain saturated, and unsaturated dicarboxylic organic acids (C6-C10). RFD rats also showed a decrease in the concentration of free LC in the blood plasma and in tissues, an increase in the level of isobutyryl- and isovalerylcarnitine in muscle tissue, and reduction in the level of acetyl- and propionylcarnitine in the blood plasma, kidney, and liver (all changes detected relative to animals in the control group). The introduction of LC to RFD rats normalized the LC homeostasis by increasing free LC concentration in the blood plasma and tissues, enhanced the acyl-LC excretion with urine and the level in tissues, and reduced the manifestations of organic acidurea.


Femtosecond pulse laser cleaning of spray paint from heritage stone surfaces.
We explore the use of femtosecond laser pulses to clean a variety of colors of spray paint from the Moruya granite, a stone with high heritage value that is widely used for monuments and sculptures in Sydney and New South Wales (Australia). The efficiency of the cleaning treatment and the effects on the stone substrate are evaluated using optical microscopy, optical profilometry, Raman spectroscopy, energy-dispersive X-ray spectroscopy, and colorimetry. We demonstrate that femtosecond laser cleans granite without damaging it and without discoloration when the laser fluence is set below the damage threshold of the stone.


Design, Synthesis and Evaluation of Pentacyclic Triterpenoids Similar to Glycyrrhetinic Acid Via Combination of Chemical and Microbial Modification as Glycogen Phosphorylases Inhibitor.
A series of pentacyclic triterpenoids similar to glycyrrhetinic acid were designed and synthesized via the combination of chemical modification and microbial catalysis. All products were screened for the glycogen phosphorylases inhibitory activities in vitro. Within this series of derivatives, compound 5 displayed good inhibitory activities with IC50 value of 27.7 μM, which is better than that of the other derivatives and glycyrrhetinic acid. Structure-activity relationship (SAR) analysis of these inhibitors was also discussed.


Thickness of elemental and binary single atomic monolayers.
The thickness of monolayers is a fundamental property of two-dimensional (2D) materials that has not found the necessary attention. It plays a crucial role in their mechanical behavior, the determination of related physical properties such as heat transfer, and especially the properties of multilayer systems. Measurements of the thickness of free-standing monolayers are widely lacking and notoriously too large. Consistent thicknesses have been reported for single layers of graphene, boronitrene, and SiC derived from interlayer spacing measured by X-ray diffraction in multilayer systems, first-principles calculations of the interlayer spacing, and tabulated van der Waals (vdW) diameters. Furthermore, the electron density-based volume model agrees with the geometric slab model for graphene and boronitrene. For other single-atom monolayers DFT calculations and molecular dynamics (MD) simulations deliver interlayer distances that are often much smaller than the vdW diameter, owing to further electrostatic and (weak) covalent interlayer interaction. Monolayers strongly bonded to a surface also show this effect. If only weak vdW forces exist, the vdW diameter delivers a reasonable thickness not only for free-standing monolayers but also for few-layer systems and adsorbed monolayers. Adding the usually known corrugation effect of buckled or puckered monolayers to the vdW diameter delivers an upper limit of the monolayer thickness. The study presents a reference database of thickness values for elemental and binary group-IV and group-V monolayers, as well as binary III-V and IV-VI compounds.


AbobotulinumtoxinA: A 25-Year History.
During the late 1960s and early 1970s, Alan Scott showed that intramuscular injections of botulinum toxin (BoNT) corrected nonaccommodative strabismus without resorting to surgery. The UK doctors who trained with Scott soon realized the significant potential offered by BoNT type A as a therapeutic option for several difficult-to-treat diseases. This led to a collaboration between these pioneering clinicians and the Centre for Applied Microbiology and Research at Porton Down, United Kingdom, and, in turn, to the development and commercialization of abobotulinumtoxinA as Dysport (Dystonia/Porton Down; Ipsen Biopharm Ltd., Wrexham, UK). Dysport was approved in Europe for the treatment of specific dystonias in December 1990 and now has marketing authorizations in 75 countries. Since then, the use of BoNT in therapeutic and aesthetic indications has grown year-on-year, and continues to expand well beyond Scott's initial aim. For example, ongoing trials are assessing potential new indications for BoNT-A, including acne and psoriasis. Furthermore, a growing number of other BoNT products, often termed "biosimilars," together with innovative formulations of well-established BoNT types, are likely to reach the market over the next few years. This review focuses on the history of Dysport to mark the 25th anniversary of its first launch in the United Kingdom.


Immediate postoperative laser resurfacing improves second intention healing on the nose: 5-year experience.
Mohs surgery defects on convex nasal surfaces do not reliably heal well by second intention. Prior to the availability of laser resurfacing we found that immediate postoperative dermabrasion improved the predicted outcome from second intention healing for these defects. To determine the ability of immediate postoperative CO2 and Er:YAG laser resurfacing to improve predicted healing by second intention. Seventy-four patients with Mohs surgical defects on the nose underwent immediate postoperative resurfacing with either a scanned CO2 or long-pulsed Er:YAG laser. Thirty patients had photographs of appropriate quality and follow-up for evaluation by a panel of nine objective physicians. All 74 patients were satisfied with their result and none have requested scar revision. Of the 30 patients evaluated by the panel, all were scored acceptable to excellent. Immediate laser resurfacing can improve the predicted outcome from second intention healing on the nose and should be considered for select patients.


[Nephropathy in a case of Fabry's disease].
The present communication describes a 22 year-old male with clinical, biochemical and histopathological signs of Fabry's disease (FD). This is the second case of FD reported in Mexico. The main clinical manifestations of this patient were during puberty: intermittent weakness, acroparesthesias and hypohidrosis. At age 21 he developed heavy proteinuria (7-12 g/day) systemic hypertension and moderate renal failure. The kidney biopsy showed the typical alterations described in FD. On light microscopy, the renal pathology showed numerous vacuolated epithelial cells in the glomerular wall, giving them a "foamy" appearance; vacuolization was due to the presence of lipid material within these cells and in the interstitium. In electron microscopy, dense, irregularly shaped osmiophilic, laminated bodies with myelinlike configuration, were seen within the glomerular epithelial cells and in the urinary spaces. The low alfa-galactosidase activity in serum (0.18 nmol/h/mL normal values: 12.8) confirmed the diagnosis. Our findings in this case indicate that patients with FD may start with renal symptoms alone, which were full blown nephrotic syndrome, hypertension and moderate renal failure.


The proteasome inhibitor bortezomib sensitizes melanoma cells toward adoptive CTL attack.
Adoptive transfer of tumor-specific cytolytic T lymphocytes (CTL) results in target cell lysis by activating the intrinsic apoptotic cell death program. Not surprisingly, deregulation of the apoptotic machinery is one of the central mechanisms by which tumor cells escape immune destruction despite specific CTL recognition. Here we show that treatment with the proteasome inhibitor bortezomib sensitizes previously resistant tumor cells for cytolytic T-cell attack. Human T cells were redirected toward melanoma cells by engineered expression of an immunoreceptor with binding specificity for high molecular weight-melanoma-associated antigen. Established melanoma cell lines as well as primary melanoma cells from tumor biopsies, which are notoriously resistant toward T-cell lysis, became sensitive upon bortezomib treatment. Detailed analysis of the underlying molecular mechanism revealed that bortezomib treatment induced mitochondrial accumulation of NOXA, which potentiated the release of mitochondrial second mitochondria-derived activator of caspase (SMAC) in response to CTL effector functions, including caspase-8 and granzyme B. Our data indicate that proteasome inhibition increases the sensitivity of tumor cells toward cytolytic T-cell attack by NOXA-mediated enhancement of mitochondrial SMAC release.