Milk mid-infrared spectral data as a tool to predict feed intake in lactating Norwegian Red dairy cows.
Mid-infrared (MIR) spectroscopy of milk was used to predict dry matter intake (DMI) and net energy intake (NEI) in 160 lactating Norwegian Red dairy cows. A total of 857 observations were used in leave-one-out cross-validation and external validation to develop and validate prediction equations using 5 different models. Predictions were performed using (multiple) linear regression, partial least squares (PLS) regression, or best linear unbiased prediction (BLUP) methods. Linear regression was implemented using just milk yield (MY) or fat, protein, and lactose concentration in milk (Mcont) or using MY together with body weight (BW) as predictors of intake. The PLS and BLUP methods were implemented using just the MIR spectral information or using the MIR together with Mcont, MY, BW, or NEI from concentrate (NEIconc). When using BLUP, the MIR spectral wavelengths were always treated as random effects, whereas Mcont, MY, BW, and NEIconc were considered to be fixed effects. Accuracy of prediction (R) was defined as the correlation between the predicted and observed feed intake test-day records. When using the linear regression method, the greatest R of predicting DMI (0.54) and NEI (0.60) in the external validation was achieved when the model included both MY and BW. When using PLS, the greatest R of predicting DMI (0.54) and NEI (0.65) in the external validation data set was achieved when using both BW and MY as predictors in combination with the MIR spectra. When using BLUP, the greatest R of predicting DMI (0.54) in the external validation was when using MY together with the MIR spectra. The greatest R of predicting NEI (0.65) in the external validation using BLUP was achieved when the model included both BW and MY in combination with the MIR spectra or when the model included both NEIconc and MY in combination with MIR spectra. However, although the linear regression coefficients of actual on predicted values for DMI and NEI were not different from unity when using PLS, they were less than unity for some of the models developed using BLUP. This study shows that MIR spectral data can be used to predict NEI as a measure of feed intake in Norwegian Red dairy cattle and that the accuracy is augmented if additional, often available data are also included in the prediction model.


Arsenite induces a cell stress-response gene, RTP801, through reactive oxygen species and transcription factors Elk-1 and CCAAT/enhancer-binding protein.
RTP801 is a newly discovered stress-response gene that is induced by hypoxia and other cell stress signals. Arsenic is a heavy metal that is linked to carcinogenesis in humans. Here, we investigated the mechanism by which arsenic induces RTP801 transcription. In HaCaT human keratinocytes, arsenite was able to induce a rapid rise in the RTP801 mRNA level. Correspondingly, arsenite treatment was capable of stimulating a 2.5 kb human RTP801 promoter. Such a stimulatory effect was inhibited by co-expression of superoxide dismutase or glutathione peroxidase, and was abrogated by N-acetylcysteine, implying that ROS (reactive oxygen species) were involved in transcriptional regulation of the RTP801 gene. A series of deletion studies with the promoter revealed a critical arsenic-responsive region between -1057 and -981 bp of the promoter. Point mutations of the putative Elk-1 site and the C/EBP (CCAAT/enhancer-binding protein) site within this region were able to reduce the stimulatory effect of arsenite, indicating that Elk-1 and C/EBP are involved in transcriptional regulation of the RTP801 gene by arsenite. Furthermore, a gel mobility-shift assay demonstrated that arsenite was able to mount the rapid formation of a protein complex that bound the arsenic-responsive region as well as the C/EBP-containing sequence. The arsenite stimulation on RTP801 transcription was partly mediated by the ERK (extracellular-signal-regulated kinase) pathway, since the effect of RTP801 was inhibited by a selective ERK inhibitor. In addition, overexpression of Elk-1 and C/EBPbeta was able to elevate the promoter activity. Therefore these studies indicate that RTP801 is a transcriptional target of arsenic in human keratinocytes, and that arsenic and ROS production are linked to Elk-1 and C/EBP in the transcriptional control.


Refinement of Multiconformer Ensemble Models from Multi-temperature X-ray Diffraction Data.
Conformational ensembles underlie all protein functions. Thus, acquiring atomic-level ensemble models that accurately represent conformational heterogeneity is vital to deepen our understanding of how proteins work. Modeling ensemble information from X-ray diffraction data has been challenging, as traditional cryo-crystallography restricts conformational variability while minimizing radiation damage. Recent advances have enabled the collection of high quality diffraction data at ambient temperatures, revealing innate conformational heterogeneity and temperature-driven changes. Here, we used diffraction datasets for Proteinase K collected at temperatures ranging from 313 to 363K to provide a tutorial for the refinement of multiconformer ensemble models. Integrating automated sampling and refinement tools with manual adjustments, we obtained multiconformer models that describe alternative backbone and sidechain conformations, their relative occupancies, and interconnections between conformers. Our models revealed extensive and diverse conformational changes across temperature, including increased bound peptide ligand occupancies, different Ca2+ binding site configurations and altered rotameric distributions. These insights emphasize the value and need for multiconformer model refinement to extract ensemble information from diffraction data and to understand ensemble-function relationships.


Results of reoperation for hip dislocation: the big picture.
Hip instability remains a costly complication of primary (3%) and revision (10%) procedures. In those with well-oriented components, instability may be anticipated in about 70% from advancement of the trochanter. Articular reorientation readily is affected by the use of modular cups with elevated rims. This has proven to be an effective strategy both to lessen the likelihood of an initial dislocation after both primary and revision procedures. It has also proven to be an effective strategy to treat the unstable implant, especially if cup orientation has been defined as the primary problem. Larger head sizes in the range of low-friction arthroplasty, such as 32 mm, are not any more stable than 22-mm diameter implants. Anatomic-sized heads as used in bipolar devices are effective in treating established instability in up to 90% of instances. The most popular current option is that of the constrained head/cup articulation. Good short-term results have revealed success in more than 90%. However, the effectiveness is design dependent, and the long-term effectiveness understandably is questioned as reports of mechanical failure begin to emerge.


A DNP-supported solid-state NMR study of carbon species in fluid catalytic cracking catalysts.
A combination of solid-state NMR techniques supported by EPR and SEM-EDX experiments was used to localize different carbon species (coke) in commercial fluid catalytic cracking catalysts. Aliphatic coke species formed during the catalytic process and aromatic coke species deposited directly from the feedstock respond differently to dynamic nuclear polarization signal enhancement in integral and crushed FCC particles, indicating that aromatic species are mostly concentrated on the outside of the catalyst particles, whereas aliphatic species are also located on the inside of the FCC particles. The comparison of solid-state NMR data with and without the DNP radical at low and ambient temperature suggests the proximity between aromatic carbon deposits and metals (mostly iron) on the catalyst surface. These findings potentially indicate that coke and iron deposit together, or that iron has a role in the formation of aromatic coke.


Maximizing Drift Is Not Optimal for Solving OneMax.
It seems very intuitive that for the maximization of the OneMax problem Om(x):=∑i=1nxi the best that an elitist unary unbiased search algorithm can do is to store a best so far solution, and to modify it with the operator that yields the best possible expected progress in function value. This assumption has been implicitly used in several empirical works. In Doerr et al. (2020), it was formally proven that this approach is indeed almost optimal. In this work, we prove that drift maximization is not optimal. More precisely, we show that for most fitness levels between n/2 and 2n/3 the optimal mutation strengths are larger than the drift-maximizing ones. This implies that the optimal RLS is more risk-affine than the variant maximizing the stepwise expected progress. We show similar results for the mutation rates of the classic (1+1) Evolutionary Algorithm (EA) and its resampling variant, the (1+1) EA>0. As a result of independent interest we show that the optimal mutation strengths, unlike the drift-maximizing ones, can be even.


Bacteroides uniformis CECT 7771 ameliorates metabolic and immunological dysfunction in mice with high-fat-diet induced obesity.
Associations have been made between obesity and reduced intestinal numbers of members of the phylum Bacteroidetes, but there is no direct evidence of the role these bacteria play in obesity. Herein, the effects of Bacteroides uniformis CECT 7771 on obesity-related metabolic and immune alterations have been evaluated. Adult (6-8 week) male wild-type C57BL-6 mice were fed a standard diet or a high-fat-diet HFD to induce obesity, supplemented or not with B. uniformis CECT 7771 for seven weeks. Animal weight was monitored and histologic, biochemical, immunocompetent cell functions, and features of the faecal microbiota were analysed after intervention. The oral administration of B. uniformis CECT 7771 reduced body weight gain, liver steatosis and liver cholesterol and triglyceride concentrations and increased small adipocyte numbers in HFD-fed mice. The strain also reduced serum cholesterol, triglyceride, glucose, insulin and leptin levels, and improved oral tolerance to glucose in HFD fed mice. The bacterial strain also reduced dietary fat absorption, as indicated by the reduced number of fat micelles detected in enterocytes. Moreover, B. uniformis CECT 7771 improved immune defence mechanisms, impaired in obesity. HFD-induced obesity led to a decrease in TNF-α production by peritoneal macrophages stimulated with LPS, conversely, the administration of B. uniformis CECT 7771 increased TNF-α production and phagocytosis. Administering this strain also increased TNF-α production by dendritic cells (DCs) in response to LPS stimulation, which was significantly reduced by HFD. B. uniformis CECT 7771 also restored the capacity of DCs to induce a T-cell proliferation response, which was impaired in obese mice. HFD induced marked changes in gut microbiota composition, which were partially restored by the intervention. Altogether, the findings indicate that administration of B. uniformis CECT 7771 ameliorates HFD-induced metabolic and immune dysfunction associated with intestinal dysbiosis in obese mice.


Treating an aged pentachlorophenol- (PCP-) contaminated soil through three sludge handling processes, anaerobic sludge digestion, post-sludge digestion and sludge land application.
The extensive pentachlorophenol (PCP) contamination and its increasing treatment costs motivate the search for a more competitive treatment alternative. In a municipal wastewater treatment plant, anaerobic sludge-handling processes comprises three bio-processes, namely the anaerobic sludge digestion, post-sludge digestion and sludge land application, which reduce sludge organic content and make sludge a good fertilizer for land application. Availability and effectiveness make the anaerobic sludge handling processes potential technologies to treat PCP-contaminated soil. The technical feasibility of using anaerobic sludge bioprocesses was studied by treating PCP soil in two pilot digesters to simulate the primary sludge digestion, in serum bottles to mimic the post-sludge digestion, and in glass pans to represent the on-site sludge application. For primary digestion, the results showed that up to 0.98 and 0.6 mM of chemical and soil PCP, respectively, were treated at nearly 100% and 97.5% efficiencies. The PCP was transformed 95% to 3-MCP, 4.5% to 3,4-DCP, and 0.5% to 3,5-DCP. For post-digestion, 100% pure chemical PCP and greater than 95% soil PCP were removed in less than 6 months with no chlorophenol residues of any kind. Complete removal of PCP by-products makes this process a good soil cleanup method. For on-site treatment, PCP was efficiently treated by multiple sludge application; however, the PCP residue was observed due to the high initial PCP content in soil. Overall, more mass PCP per unit sludge per day was processed using the primary sludge digestion than the on-site soil treatment or post-sludge digestion. And, sludge acclimation resulted in better PCP treatment efficiencies with all three processes.


CD164 regulates the tumorigenesis of ovarian surface epithelial cells through the SDF-1α/CXCR4 axis.
CD164 (endolyn), a sialomucin, has been reported to play a role in the proliferation, adhesion, and differentiation of hematopoietic stem cells. The potential association of CD164 with tumorigenicity remains unclear. The clinicopathological correlation of ovarian cancer with CD164 was assessed in a 97-patient tumor tissue microarray. Overexpression or silence CD164 was to analyze the effect of CD164 on the proliferation, colony formation and apoptosis via a mouse xenograft and western blotting analysis. The subcellular localization of CD164 was collected in the immunohistochemical and confocal analysis. Our data demonstrated that higher expression levels of CD164 were identified in malignant ovarian cancer cell lines, such as SKOV3 and HeyA8. The clinicopathological correlation analysis showed that the upregulation of CD164 protein was significantly associated with tumor grade and metastasis. The overexpression of CD164 in human ovarian epithelial surface cells promoted cellular proliferation and colony formation and suppressed apoptosis. These tumorigenicity effects of CD164 were reconfirmed in a mouse xenograft model. We also found that the overexpression of CD164 proteins increased the amounts of CXCR4 and SDF-1α and activated the SDF-1α/CXCR4 axis, inducing colony and sphere formation. Finally, we identified the subcellular localization of CD164 in the nucleus and cytosol and found that nuclear CD164 might be involved in the regulation of the activity of the CXCR4 promoter. Our findings suggest that the increased expression of CD164 is involved in ovarian cancer progression via the SDF-1α/CXCR4 axis, which promotes tumorigenicity. Thus, targeting CD164 may serve as a potential ovarian cancer biomarker, and targeting CD164 may serve as a therapeutic modality in the management of high-grade ovarian tumors.


Strain- and age-related alteration of proteins in the brain of SAMP8 and SAMR1 mice.
In order to discover and identify the key protein biomarkers in the aging process, we performed a differential proteomic analysis of hippocampus and cortex in 5- and 15-month old senescence-accelerated mouse prone 8 (SAMP8) as well as in control strain SAM/resistant 1 (SAMR1). Using 2-DE combined with MALDI TOF/TOF mass spectrometry, about 1700 protein spots were isolated, and three groups of differentially expressed proteins were identified. The first group contained the strain-specific and non-age-related differential proteins that were differentially expressed in SAMP8 compared with SAMR1 mice. The changes might be implicated in the genetic difference between SAMP8 and SAMR1 mice; specifically, the proteins ubiquitin carboxyl-terminal esterase L3, mitofilin, adenylate kinase 4, and an unnamed protein product (gi|12847201). The proteins in the second group were age-specific, which were differentially expressed between 5- and 15-month old SAM mice. Those proteins are particularly interesting since the changes were aging-related and some of them were previously reported to be expressed in Alzheimer's disease patients. These proteins included N-myc downstream regulated gene 2, enolase 2, Cu/Zn superoxide dismutase, myosin, and two unnamed protein products (gi|74214304 and gi|74178239). The protein in the third group was SAMP8 specific-age-related protein, which was identified as heme binding protein 1. The present study provides new information about SAMP8 specific and aging-related protein changes in brain. Further investigations will be performed to reveal the significance of these proteins in brain aging process and the potential roles as biomarkers for effective diagnosis and therapy.


Honeycomb blocks composed of carbonate apatite, β-tricalcium phosphate, and hydroxyapatite for bone regeneration: effects of composition on biological responses.
Synthetic scaffolds exhibiting bone repair ability equal to that of autogenous bone are required in the fields of orthopedics and dentistry. A suitable synthetic bone graft substitute should induce osteogenic differentiation of mesenchymal stem cells, osteogenesis, and angiogenesis. In this study, three types of honeycomb blocks (HCBs), composed of hydroxyapatite (HAp), β-tricalcium phosphate (TCP), and carbonate apatite (CO3Ap), were fabricated, and the effects of HCB composition on bone formation and maturation were investigated. The HC structure was selected to promote cell penetration and tissue ingrowth. HAp and β-TCP HCBs were fabricated by extrusion molding followed by sintering. The CO3Ap HCBs were fabricated by extrusion molding followed by sintering and dissolution-precipitation reactions. These HCBs had similar macroporous structures: all harbored uniformly distributed macropores (∼160 μm) that were regularly arrayed and penetrated the blocks unidirectionally. Moreover, the volumes of macropores were nearly equal (∼0.15 cm3/g). The compressive strengths of CO3Ap, HAp, and β-TCP HCBs were 22.8 ± 3.5, 34.2 ± 3.3, and 24.4 ± 2.4 MPa, respectively. Owing to the honeycomb-type macroporous structure, the compressive strengths of these HCBs were higher than those of commercial scaffolds with intricate three-dimensional or unidirectional macroporous structure. Notably, bone maturation was markedly faster in CO3Ap HCB grafting than in β-TCP and HAp HCB grafting, and the mature bone area percentages for CO3Ap HCBs at postsurgery weeks 4 and 12 were 14.3- and 4.3-fold higher and 7.5- and 1.4-fold higher than those for HAp and β-TCP HCBs, respectively. The differences in bone maturation and formation were probably caused by the disparity in concentrations of calcium ions surrounding the HCBs, which were dictated by the inherent material resorption behavior and mechanism; generally, CO3Ap is resorbed only by osteoclastic resorption, HAp is not resorbed, and β-TCP is rapidly dissolved even in the absence of osteoclasts. Besides the composition, the microporous structure of HC struts, inevitably generated during the formation of HCBs of various compositions, may contribute to the differences in bone maturation and formation.


A toxicological and dermatological assessment of alkyl cyclic ketones when used as fragrance ingredients. RIFM Expert Panel.
The alkyl cyclic ketone (ACK) fragrance ingredients are a diverse group of structures with similar metabolic and toxicity profiles. ACK fragrance materials demonstrate low acute toxicity. Upon repeat dose testing, some adverse effects in biochemical and hematological parameters, and slightly increased liver and kidney weights were reported, primarily at high doses, resulting from adaptive effects. Developmental effects occurred only in the presence of maternal toxicity. Assays in bacteria and mammalian cell systems and the mouse micronucleus assay did not demonstrate genotoxicity. ACK fragrance ingredients are considered non-irritating to the skin of humans; results showed few reactions, most of which were equivocal or involved doses greater than those in consumer products. Mild to moderate eye irritation in animal tests was observed with most compounds; however, full recovery was usually observed. Human sensitization studies indicate that ACK fragrance ingredients have a low sensitization potential. Diagnostic patch-tests indicated low sensitizing potential in humans; except for fragrance materials which caused reactions at 1% or 5%. Phototoxicity and photosensitization were not demonstrated in humans, and, with the possible exception of acetyl cedrene, would not be expected. It is concluded that ACK materials do not present a safety concern at current levels of use as fragrance ingredients.


Predictive validity of preschool screening tools for language and behavioural difficulties: A PRISMA systematic review.
Preschool screening for developmental difficulties is increasingly becoming part of routine health service provision and yet the scope and validity of tools used within these screening assessments is variable. The aim of this review is to report on the predictive validity of preschool screening tools for language and behaviour difficulties used in a community setting. Studies reporting the predictive validity of language or behaviour screening tools in the preschool years were identified through literature searches of Ovid Medline, Embase, EBSCO CINAHL, PsycInfo and ERIC. We selected peer-reviewed journal articles reporting the use of a screening tool for language or behaviour in a population-based sample of children aged 2-6 years of age, including a validated comparison diagnostic assessment and follow-up assessment for calculation of predictive validity. A total of eleven eligible studies was identified. Six studies reported language screening tools, two reported behaviour screening tools and three reported combined language & behaviour screening tools. The Language Development Survey (LDS) administered at age 2 years achieved the best predictive validity performance of the language screening tools (sens 67%, spec 94%, NPV 88% and PPV 80%). The Strengths and Difficulties Questionnaire (SDQ) administered at age 4 years achieved the best predictive validity compared to other behaviour screening tools (Sens 31%, spec 93%, NPV 84% and PPV 52%). The SDQ and Sure Start Language Measure (SSLM) administered at 2.5 years achieved the best predictive validity of the combined language & behaviour assessments (sens 87%, spec 64%, NPV 97% and PPV 31). Predictive validity data and diagnostic odds ratios identified language screening tools as more effective and achieving higher sensitivity and positive predictive value than either behaviour or combined screening tools. Screening tools with combined behaviour and language assessments were more specific and achieved higher negative predictive value than individual language or behaviour screening tools. Parent-report screening tools for language achieved higher sensitivity, specificity and negative predictive value than direct child assessment. Universal screening tools for language and behaviour concerns in preschool aged children used in a community setting can demonstrate excellent predictive validity, particularly when they utilise a parent-report assessment. Incorporating these tools into routine child health surveillance could improve the rate of early identification of language and behavioural difficulties, enabling more informed referrals to specialist services and facilitating access to early intervention.


The immunology of multiple sclerosis.
Our incomplete understanding of the causes and pathways involved in the onset and progression of multiple sclerosis (MS) limits our ability to effectively treat this complex neurological disease. Recent studies explore the role of immune cells at different stages of MS and how they interact with cells of the central nervous system (CNS). The findings presented here begin to question the exclusivity of an antigen-specific cause and highlight how seemingly distinct immune cell types can share common functions that drive disease. Innovative techniques further expose new disease-associated immune cell populations and reinforce how environmental context is critical to their phenotype and subsequent role in disease. Importantly, the differentiation of immune cells into a pathogenic state is potentially reversible through therapeutic manipulation. As such, understanding the mechanisms that provide plasticity to causal cell types is likely key to uncoupling these disease processes and may identify novel therapeutic targets that replace the need for cell ablation.


First Report of Smut of Saccharum arundinaceum Caused by Sporisorium sorghi in Yunnan, China.
Saccharum arundinaceum (Retz.) is a wild species of S. officinarum with good resistance to pests, diseases, drought, cold, and infertile soils, and has favorable tillering potential (4). Hybridization of S. arundinaceum with sugarcane may enable favorable characteristics of S. arundinaceum to be exploited, broadening the genetic base of sugarcane breeding. In May 2009, symptoms of a disease were observed on plants of S. arundinaceum at the National Nursery of Sugarcane Germplasm Resources in Yunnan Province, China, on approximately 10% of 120 plants within three germplasm collections of S. arundinaceum. The initial symptom was dwarfing, resulting in the spike heading 1 to 2 months earlier than on healthy plants. As infection progressed, the spikelet of each diseased plant became full of the black, powdery mass of fungal spores enclosed in an off-white membrane. Finally, the entire spike became severely infected. A Sporisorium sp. was isolated from diseased spike tissues that were surface-sterilized with 70% alcohol, then with 0.1% mercury chloride, dipped in sterilized water three times for 3 min each time, dried on sterilized absorbent paper, and placed onto potato dextrose agar (PDA) at 27 ± 2°C in the dark. Nine fungal isolates were identified as Sporisorium sorghi Ehrenberg ex. Link based on sorus morphology as well as spore morphology and color (1). Sori were cylindrical or oval and 2.5 to 12.0 mm in diameter. Spores were spherical, light olive-brown or black, and 5.0 to 9.0 × 4.0 to 8.5 mm. Preliminary morphological identification of the fungus was confirmed by PCR assay using genomic DNA extracted from the mycelia of pure cultures of each of nine isolates, which generated a 750-bp amplified region of the internal transcribed spacer (ITS) region of rDNA using ITS1/ITS4 universal primers (3). The ITS region was then sequenced (GenBank Accession No. JX183795), and displayed 98% similarity with the ITS sequence of an isolate of S. sorghi from Sorghum bicolor from each of France (AF038828.1) and Germany (AY740021.1). A pathogenicity test was completed with the S. arundinaceum isolate of S. sorghi by spraying 20 ml of a spore suspension (104 conidia/ml) onto each of 10 stems (2) of S. arundinaceum in a field in February 2010. The spore suspension was prepared from 30-day-old cultures growing on PDA. Stems were inoculated approximately 4 cm above the ground, where each stem was cut with sterilized shears. For the control treatment, each of 10 cut stems was inoculated with 20 ml of sterilized water. Three replications of 10 stems were used for each treatment. Approximately 3.5 months after inoculation, symptoms began to appear on 18 of the 30 stems (three replicates of 10 stems) inoculated with the S. sorghi spore suspension, when heading occurred on the stems. Five months later, each entire spike exhibited symptoms. No symptoms were observed on control spikes of stems inoculated with water. S. sorghi was reisolated onto PDA from the tissues of diseased spikes on inoculated stems, and the reisolates were identified by PCR assay using ITS1/ITS4 universal primers, as described above. S. sorghi was not isolated from the spike tissues of the control plants. To our knowledge, this is the first report of smut on S. arundinaceus caused by S. sorghi in Yunnan Province, China. References: (1) L. Guo. Flora Fungorum Sinicorum.12:51, 2000. (2) W. F. Li et al. Chinese Plant Prot. 34:127, 2008. (3) W. J. Lin et al. Chinese Agri. Sci. Bull. 23:293, 2007. (4) X. Lu et al. Southwest China J. Agric. Sci. 20:103, 2007.


Capture and release of proteins on the nanoscale by stimuli-responsive elastin-like polypeptide "switches".
This article describes the fabrication and characterization of stimulus-responsive elastin-like polypeptide (ELP) nanostructures grafted onto omega-substituted thiolates that were patterned onto gold surfaces by dip-pen nanolithography (DPN). In response to external stimuli such as changes in temperature or ionic strength, ELPs undergo a switchable and reversible, hydrophilic-hydrophobic phase transition at a lower critical solution temperature (LCST). We exploited this phase transition behavior to reversibly immobilize a thioredoxin-ELP (Trx-ELP) fusion protein onto the ELP nanopattern above the LCST. Subsequent binding of an anti-thioredoxin monoclonal antibody (anti-Trx) to the surface-captured thioredoxin showed the presentation of the immobilized protein in a sterically accessible orientation in the nanoarray. We also showed that the resulting Trx-ELP/anti-Trx complex formed above the LCST could be reversibly dissociated below the LCST. These results demonstrate the intriguing potential of ELP nanostructures as generic, reversible, biomolecular switches for on-chip capture and release of a small number (order 100-200) of protein molecules in integrated, nanoscale bioanalytical devices. We also investigated the molecular mechanism underlying this switch by measuring the height changes that accompany the binding and desorption steps and by adhesion force spectroscopy using atomic force microscopy.


Early reactions to cannabis predict later dependence.
While there is a growing literature on the linkages between early subjective responses to nicotine and alcohol and later risks of nicotine or alcohol dependence, to date there has been no study of this issue in relation to cannabis. To examine the extent to which subjective responses to early (prior to the age of 16 years) cannabis use were associated with subsequent cannabis dependence in a birth cohort studied to the age of 21 years. Data on early (prior to the age of 16 years) subjective reactions to cannabis use and subsequent cannabis dependence were gathered over the course of the Christchurch Health and Development Study, a 21-year longitudinal study of a birth cohort of children born in Christchurch, New Zealand. General community sample. Members of a population-based birth cohort (86.5% white, 11.3% New Zealand Maori, and 2.2% Pacific Island). Main Outcome Measure Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition cannabis dependence (for those aged 16-21 years). Of the cohort, 198 (20%) had used cannabis prior to the age of 16 years. Among this high-risk group, rates of dependence were high with 21.7% meeting DSM-IV criteria for cannabis dependence by the age of 21 years. There were clear tendencies for rates of cannabis dependence to increase with increasing reports of positive responses to early cannabis use: those reporting 5 positive responses had odds of cannabis dependence that were 28.5 (95% confidence interval, 6.3-133.8) times higher than those not reporting positive reactions to cannabis. The association held (odds ratio, 23.4; 95% confidence interval, 4.0-135.9) after control for potentially confounding factors including the extent of use of cannabis prior to age 16 years. The extent of early negative reactions to cannabis was unrelated to later cannabis dependence. Early subjective responses to cannabis are prognostic of later cannabis dependence. These findings may suggest the presence of genetically mediated individual differences in early responsiveness to cannabis. Clinicians should be aware that young people who report positive reactions to early use of cannabis are at increased risks of later cannabis dependence.


A novel approach to propagation pattern analysis in intracardiac atrial fibrillation signals.
The purpose of this study is to investigate propagation patterns in intracardiac signals recorded during atrial fibrillation (AF) using an approach based on partial directed coherence (PDC), which evaluates directional coupling between multiple signals in the frequency domain. The PDC is evaluated at the dominant frequency of AF signals and tested for significance using a surrogate data procedure specifically designed to assess causality. For significantly coupled sites, the approach allows also to estimate the delay in propagation. The methods potential is illustrated with two simulation scenarios based on a detailed ionic model of the human atrial myocyte as well as with real data recordings, selected to present typical propagation mechanisms and recording situations in atrial tachyarrhythmias. In both simulation scenarios the significant PDCs correctly reflect the direction of coupling and thus the propagation between all recording sites. In the real data recordings, clear propagation patterns are identified which agree with previous clinical observations. Thus, the results illustrate the ability of the novel approach to identify propagation patterns from intracardiac signals during AF, which can provide important information about the underlying AF mechanisms, potentially improving the planning and outcome of arrhythmia ablation.


The relationship of micronucleus frequency and nuclear division index with coronary artery disease SYNTAX and Gensini scores.
We aimed to evaluate the relationship of micronucleus (MN) frequency and nuclear division index (NDI) with SYNTAX and Gensini scores and thrombolysis in myocardial infarction (TIMI) frame counts of coronary arteries in patients undergoing coronary angiography. In a single-center prospective observational study, a total of 63 individuals, 48 consecutive patients with coronary artery disease (CAD) and 15 healthy people were included. Before coronary angiography (exposure to X-ray), blood samples were collected for lymphocyte cultures, MN and NDI measurements. According to the SYNTAX and Gensini scores, patients were allocated into two groups. Group 1 and 2 included the patients with SYNTAX scores <22 and ≥22 points, respectively. Similarly, groups according to Gensini scores included the ones <23 and ≥23 points. MN test was used for in vitro studies in human peripheral lymphocytes. Binucleated lymphocytes were calculated for each patient. MN frequency was significantly higher in group 2 than group 1 and in group 1 than control group (p<0.001). NDI was significantly higher in control group than group 1 and in group 1 than group 2 (p=0.003). MN frequency had positive but moderate correlation with SYNTAX and Gensini scores and TFCs of left anterior descending (LAD), circumflex and right coronary arteries (r=0.394, p=0.003; r=0.458, p<0.001; r=0.425, p<0.001; r=0.469, p<0.001; and r=0.475, p<0.001, respectively). We can conclude that as the degree of atherosclerosis increases and coronary flow worsens, MN frequency increases and NDI decreases. Our results may help to elucidate the relationship of DNA damage in pathophysiology of atherosclerosis and endothelial dysfunction in patients with stable CAD.


Oral clofarabine in the treatment of patients with higher-risk myelodysplastic syndrome.
Efficacy and toxicity profile of orally administered clofarabine were evaluated in patients with higher-risk myelodysplastic syndrome (MDS). Thirty-two patients were treated, of whom 22 had intermediate-2 or high-risk disease (International Prognostic Scoring System). Median age was 70 years (range, 53 to 86), nine patients had secondary MDS, and 20 patients experienced prior therapy failure with hypomethylating agents. Three doses of clofarabine were evaluated: 40 mg/m(2), 30 mg/m(2), and 20 mg/m(2) daily for 5 days. Courses were repeated every 4 to 8 weeks. Eight patients (25%) achieved complete remission (CR), three had (9%) hematologic improvement (HI), and three had (9%) clinical benefit (CB; overall response rate, 43%). Responses in patients who experience treatment failure with hypomethylating agents included CR in two (10%), HI in two (10%), and CB in two patients (10%). No patients died within 6 weeks of induction. Renal failure occurred in four patients in the context of myelosuppression-associated infectious complications. Common adverse events were gastrointestinal and hepatic. Myelosuppression was common, but prolonged myelosuppression (> 42 days) was rare. The toxicity profile was better with lower doses of clofarabine, whereas response rates did not differ significantly. Oral clofarabine has achieved a response rate of 43% in patients with higher-risk MDS. The optimal dose and schedule and the appropriate patient population for such therapy remain to be further defined.


A mutagenesis-derived broad-spectrum disease resistance locus in wheat.
Wheat leaf rust, stem rust, stripe rust, and powdery mildew caused by the fungal pathogens Puccinia triticina, P. graminis f. sp. tritici, P. striiformis f. sp. tritici, and Blumeria graminis f. sp. tritici, respectively, are destructive diseases of wheat worldwide. Breeding durable disease resistance cultivars rely largely on continually introgressing new resistance genes, especially the genes with different defense mechanisms, into adapted varieties. Here, we describe a new resistance gene obtained by mutagenesis. The mutant, MNR220 (mutagenesis-derived new resistance), enhances resistance to three rusts and powdery mildew, with the characteristics of delayed disease development at the seedling stage and completed resistance at the adult plant stage. Genetic analysis demonstrated that the resistance in MNR220 is conferred by a single semidominant gene mapped on the short arm of chromosome 2B. Gene expression profiling of several pathogenesis-related genes indicated that MNR220 has an elevated and rapid pathogen-induced response. In addition to its potential use in breeding for resistance to multiple diseases, high-resolution mapping and cloning of the disease resistance locus in MNR220 may lead to a better understanding of the regulation of defense responses in wheat.


Genome organization and reverse genetic analysis of a type I feline coronavirus.
In this study we report the complete sequence and genome organization of the serotype I feline coronavirus (FCoV) strain Black. Furthermore, a reverse genetic system was established for this FCoV strain by cloning a full-length cDNA copy into vaccinia virus. This clone served as basis for the generation of recombinant FCoV (recFCoV) that was shown to bear the same features in vitro as the parental FCoV. Using this system, accessory 3abc genes in the FCoV genome were replaced by green fluorescent protein (recFCoV-GFP) and Renilla luciferase genes (recFCoV-RL). In addition, we showed that feline CD14(+) blood monocytes and dendritic cells can be easily detected after infection with recFCoV-GFP. Thus, our established reverse genetic system provides a suitable tool to study the molecular biology of serotype I FCoV.


Influence of immediate loading on healing of implants installed with different insertion torques--an experimental study in dogs.
To evaluate the influence of different insertion torques on healing of implants loaded immediately or left unloaded. In six Labrador dogs, all mandibular premolars and molars were extracted. After 4 months of healing, flaps were elevated, and two implant sites were prepared at each side of the mandible. The distal sites were prepared conventionally while the mesial sites were underprepared by 0.3 mm. As a consequence, different final insertion torques of about 30 Ncm at the distal and >70 Ncm at the mesial sites were recorded. Healing abutments were applied to the left and transmucosal abutments to the right side. Flaps were sutured, crown preparation of the upper right second and third premolars was performed, and impressions were taken. Within 24 h, crowns were cemented both to implants and teeth in the right side of the mouth. After 4 months, the animals were sacrificed and ground sections obtained for histological evaluation. A higher buccal bony crestal resorption and a more apical position of the coronal level of osseointegration were found at the loaded compared with the unloaded sites. MBIC% and percentages of peri-implant mineralized tissue (MB%) were higher at the loaded compared with the unloaded sites. Moreover, a higher MBIC% was found at the lower compared with the higher final insertion torque. Immediate loading does not seem to have a negative effect on osseointegration. High torque values for the immediate loading procedures were not necessary. Probably, low torque values, were sufficient to obtain primary stability and hence may provide better osseointegration than high torque value.


Influence of Calcium and Vitamin D Intakes on Body Composition in Children and Adolescents.
Identifying nutritional strategies to maintain a healthy body weight and reduce the comorbidities associated with obesity is extremely important. We aimed to investigate whether calcium and vitamin D intakes are associated with body composition measurements in a population of children and adolescents. A cross-sectional study was conducted involving 1,060 children and adolescents (65.8% females; 34.2% males) aged 9 to 19 years. Fat mass, percentage of fat mass, and fat-free mass were measured using a body composition analyzer (TANITA BC-418MA®). The mean dietary calcium and vitamin intakes were 829.66 ± 328.34 mg/day and 200.78 ± 400.91 IU/day. Linear regression analysis revealed a lack of significant association between daily calcium and vitamin D intakes and body composition measurements, after adjusting the model for age, sex, maturation status, and energy intake. Dietary calcium and vitamin D intakes do not appear to be associated with higher adiposity measurements in children and adolescents.


Long-Chain Fatty Acyl Coenzyme A Ligase FadD2 Mediates Intrinsic Pyrazinamide Resistance in Mycobacterium tuberculosis.
Pyrazinamide (PZA) is a first-line tuberculosis (TB) drug that has been in clinical use for 60 years yet still has an unresolved mechanism of action. Based upon the observation that the minimum concentration of PZA required to inhibit the growth of Mycobacterium tuberculosis is approximately 1,000-fold higher than that of other first-line drugs, we hypothesized that M. tuberculosis expresses factors that mediate intrinsic resistance to PZA. To identify genes associated with intrinsic PZA resistance, a library of transposon-mutagenized Mycobacterium bovis BCG strains was screened for strains showing hypersusceptibility to the active form of PZA, pyrazinoic acid (POA). Disruption of the long-chain fatty acyl coenzyme A (CoA) ligase FadD2 enhanced POA susceptibility by 16-fold on agar medium, and the wild-type level of susceptibility was restored upon expression of fadD2 from an integrating mycobacterial vector. Consistent with the recent observation that POA perturbs mycobacterial CoA metabolism, the fadD2 mutant strain was more vulnerable to POA-mediated CoA depletion than the wild-type strain. Ectopic expression of the M. tuberculosis pyrazinamidase PncA, necessary for conversion of PZA to POA, in the fadD2 transposon insertion mutant conferred at least a 16-fold increase in PZA susceptibility under active growth conditions in liquid culture at neutral pH. Importantly, deletion of fadD2 in M. tuberculosis strain H37Rv also resulted in enhanced susceptibility to POA. These results indicate that FadD2 is associated with intrinsic PZA and POA resistance and provide a proof of concept for the target-based potentiation of PZA activity in M. tuberculosis.


Biosynthesis of Squalene from Farnesyl Diphosphate in Bacteria: Three Steps Catalyzed by Three Enzymes.
Squalene (SQ) is an intermediate in the biosynthesis of sterols in eukaryotes and a few bacteria and of hopanoids in bacteria where they promote membrane stability and the formation of lipid rafts in their hosts. The genes for hopanoid biosynthesis are typically located on clusters that consist of four highly conserved genes-hpnC, hpnD, hpnE, and hpnF-for conversion of farnesyl diphosphate (FPP) to hopene or related pentacyclic metabolites. While hpnF is known to encode a squalene cyclase, the functions for hpnC, hpnD, and hpnE are not rigorously established. The hpnC, hpnD, and hpnE genes from Zymomonas mobilis and Rhodopseudomonas palustris were cloned into Escherichia coli, a bacterium that does not contain genes homologous to hpnC, hpnD, and hpnE, and their functions were established in vitro and in vivo. HpnD catalyzes formation of presqualene diphosphate (PSPP) from two molecules of FPP; HpnC converts PSPP to hydroxysqualene (HSQ); and HpnE, a member of the amine oxidoreductase family, reduces HSQ to SQ. Collectively the reactions catalyzed by these three enzymes constitute a new pathway for biosynthesis of SQ in bacteria.


What do cancer patients mean when they complain of concentration and memory problems?
Cognitive function items are increasingly included in quality of life measures, and complaints of concentration and memory difficulties are often reported by cancer patients. The aim of this study was to examine the factors influencing patients' level of complaint by comparing subjective reports with objective test performance of a sample of adult lymphoma patients, disease-free and > or = 6 months after treatment. There was no significant difference between complainers and non-complainers in sociodemographic or clinical characteristics or in their performance on standard neuropsychometric tests of concentration and memory. Those reporting concentration and memory difficulties had significantly higher scores on measures of anxiety, depression and fatigue. This calls into question the validity of including cognitive function items in self-report quality of life measures. Patients who report concentration and memory difficulties should be screened for clinically significant and potentially remediable mood disorder. Objective testing remains the method of choice for assessing higher mental function.


Mechanism of the synergistic end-product regulation of Bacillus subtilis glutamine phosphoribosylpyrophosphate amidotransferase by nucleotides.
De novo purine nucleotide synthesis is regulated, at least in part, by end-product inhibition of glutamine PRPP amidotransferase. An important feature of this inhibition is the fact that certain synergistic nucleotide pairs give more than additive inhibition. The physiological importance of synergism is in amplifying regulation by the adenine and guanine nucleotide end products of de novo synthesis. Using a new method to quantitate synergism, ADP plus GMP were confirmed [Meyer, E., and Switzer, R. L. (1978) J. Biol. Chem. 254, 5397-5402] to give strong synergistic inhibition of Bacillus subtilis glutamine PRPP amidotransferase. An X-ray structure of the ternary enzyme.ADP.GMP complex established that ADP binds to the allosteric A site and GMP to the catalytic C site. GMP increased the binding affinity of ADP for the A site by approximately 20-fold. Synergism results from a specific nucleotide-nucleotide interaction that is dependent upon a nucleoside diphosphate in the A site and a nucleoside monophosphate in the C site. Furthermore, synergism is enhanced by the competition between nucleotide inhibitor and PRPP substrate for the C site. Purine base specificity results from a backbone carbonyl interaction of Lys305' with the 6-NH2 group of adenine in the A site and a Ser347 Ogamma interaction with the 2-NH2 group of guanine in the C site. Steric considerations favor binding of the nucleoside diphosphate to the A site. Site-directed replacements of key residues increased the nucleotide concentrations needed for 50% inhibition and in some cases perturbed synergism. Mutations in either of the nucleotide sites perturbed function at both sites, supporting the important role of synergism.


Bushmeat hunting and extinction risk to the world's mammals.
Terrestrial mammals are experiencing a massive collapse in their population sizes and geographical ranges around the world, but many of the drivers, patterns and consequences of this decline remain poorly understood. Here we provide an analysis showing that bushmeat hunting for mostly food and medicinal products is driving a global crisis whereby 301 terrestrial mammal species are threatened with extinction. Nearly all of these threatened species occur in developing countries where major coexisting threats include deforestation, agricultural expansion, human encroachment and competition with livestock. The unrelenting decline of mammals suggests many vital ecological and socio-economic services that these species provide will be lost, potentially changing ecosystems irrevocably. We discuss options and current obstacles to achieving effective conservation, alongside consequences of failure to stem such anthropogenic mammalian extirpation. We propose a multi-pronged conservation strategy to help save threatened mammals from immediate extinction and avoid a collapse of food security for hundreds of millions of people.


Influence of blood contamination on bond strength of a self-etching system.
To detect the influence of blood contamination (BC) on the bond strength (BS) of a self-etching bonding system (SES) to enamel and dentine. 25 human molars were longitudinally sectioned on the mesio-distal axis in order to obtain 50 specimens, which were embedded in acrylic resin. At first, the specimens were ground to expose a flat surface of enamel, and a bond strength test was performed. Afterwards, the samples were ground again in order to obtain a flat surface of dentine. Ten groups (total: n=100) were assigned according to substrate (enamel and dentine), step in the bonding sequence when contamination occurred (before the acidic primer and after the bonding resin), and contamination treatment (dry or rinse and dry procedure). Fresh human blood was introduced either before or after SES application (Clearfil SE Bond) and treated with air drying, or by rinsing and drying following application. Composite resin (Filtek Z-250,3M ESPE) was applied as inverted, truncated cured cones that were debonded in tension. The mean tensile BS values (MPa) for enamel/dentine were 19.4/23.0 and 17.1/10.0 for rinse-and-dry treatment (contamination before and after SES, respectively); while the measurements for the dry treatment, 16.2/23.3 and 0.0/0.0 contamination before and after SES, respectively. It was determined that blood contamination impaired adhesion to enamel and dentine when it occurred after bond light curing. Among the tested contamination treatments, the rinse-and-dry treatment produced the highest bond strength with BC after SES application, but it was not sufficient to recover the BS in the contamination-free group.


Left to right atrial shunting in tricuspid atresia.
In tricuspid atresia, an obligatory right to left shunt occurs at the atrial level. We have observed several patients with left to right interatrial shunts. Data from cardiac catheterisation in 40 consecutive patients were reviewed to determine the frequency and mechanism of left to right shunting in tricuspid atresia. An increase of 6% or more in oxygen saturation between the superior vena cava and the right atrium in two or more sets of saturations, representing a left to right shunt, was present in 29 out of 50 (58%) catheterisations in which the data were adequate. In most, the shunt was also seen cineangiographically in the laevophase. In only two catheterisations was an anatomical cause (ostium primum atrial septal defect in one and anomalous pulmonary venous return in the other) found. In the remaining 27 catheterisations, no anatomical cause was found. Age, Qp:Qs, and mean atrial pressure difference were similar between the shunt and non-shunt groups. In the shunt group right atrial "a" waves were equal to or higher than left atrial "a" waves and left atrial "v" waves were equal to or higher than right atrial "v" waves. Simultaneous pressure recordings (in one patient with left to right atrial shunt) from the left atrium and right atrium with isosensitised miniature pressure transducers mounted 5 cm apart showed (1) a higher pressure in the right atrium than in the left atrium during atrial systole and (2) a higher pressure in the left atrium than in the right atrium during atrial disatole. It is concluded that (a) left to right shunt across the atrial septum occurs frequently in tricuspid atresia and (b) the left to right shunt is the result of instantaneous pressure differences between the atria.


Cellular scent of influenza virus infection.
Volatile organic compounds (VOCs) emanating from humans have the potential to revolutionize non-invasive diagnostics. Yet, little is known about how these compounds are generated by complex biological systems, and even less is known about how these compounds are reflective of a particular physiological state. In this proof-of-concept study, we examined VOCs produced directly at the cellular level from B lymphoblastoid cells upon infection with three live influenza virus subtypes: H9N2 (avian), H6N2 (avian), and H1N1 (human). Using a single cell line helped to alleviate some of the complexity and variability when studying VOC production by an entire organism, and it allowed us to discern marked differences in VOC production upon infection of the cells. The patterns of VOCs produced in response to infection were unique for each virus subtype, while several other non-specific VOCs were produced after infections with all three strains. Also, there was a specific time course of VOC release post infection. Among emitted VOCs, production of esters and other oxygenated compounds was particularly notable, and these may be attributed to increased oxidative stress resulting from infection. Elucidating VOC signatures that result from the host cells response to infection may yield an avenue for non-invasive diagnostics and therapy of influenza and other viral infections.


Pulmonary thrombosis in the mouse following intravenous administration of quantum dot-labeled mesenchymal cells.
Quantum dots (QDs) are emerging as novel diagnostic agents. Yet, only a few studies have examined the possible deleterious effects of QD-labeled stem cells. We assessed the potential toxic effects of QD-labeled human embryonic palatal mesenchymal (QD-HEPM) cells in male NOD/SCID mice for six months, following the administration of a single intravenous injection. Control animals were administered with non-labeled HEPM cells. No treatment-related clinical signs, hematological, or biochemical parameters were found in the QD-HEPM animals in comparison to control animals. Histologically, multifocal organizing thrombi were noted in the pulmonary arteries of all QD-HEPM animals from the one-week study group and in one animal from the one-month group. Additionally, increased severity of perivascular inflammation was noted at the injection sites of QD-HEPM animals from the one-week group. This is the first study reporting histopathological evidence for pro-thrombotic adverse effects mediated by QD labeling.


Expression of Endometrial Receptivity Genes Increase After Myomectomy of Intramural Leiomyomas not Distorting the Endometrial Cavity.
This study was designed to investigate whether endometrial receptivity genes are altered in infertile patients with intramural leiomyomas (IM) not distorting the endometrial cavity undergoing myomectomy. We measured endometrial HOXA-10, HOXA-11, LIF, ITGB3, and ITGAV messenger RNA (mRNA) expressions levels before and after myomectomy/metroplasty during mid-luteal phase in participants with IM, submucosal leiomyomas (SM), and septate uterus and fertile participants without fibroids. Initial endometrial sampling was obtained at the time of surgery, and second sampling was obtained 3 months after myomectomy/metroplasty. Expressions of each gene were evaluated using real-time reverse transcriptase polymerase chain reaction (RT-PCR). A trend toward decreased endometrial HOXA-10, HOXA-11, and ITGAV mRNA expression was detected in both SM and IM groups before myomectomy when compared to both fertile group and septate uterus. However, the differences failed to show statistical significance. After myomectomy of IM, we have detected 12.8-fold increase in endometrial HOXA-10 mRNA expression and 9.0-fold increase in endometrial HOXA-11 mRNA expression. This increase in endometrial HOXA-10 and 11 mRNA expression was significant. Accordingly, 2 patients having intramural fibroids greater than 5 cm were able to remain pregnant after myomectomy. Conversely, submucosal myomectomy did not cause any significant effect on endometrial receptivity markers. Likewise, all markers of endometrial receptivity remained unchanged after metroplasty. Myomectomy of IM have favorable effect on endometrial HOXA-10 and 11 mRNA expression.


Coagulation and fibrinolytic profile of paediatric patients undergoing cardiopulmonary bypass.
The haemostatic system and the use of heparin during cardiopulmonary bypass (CPB) have been studied extensively in adults but not in children. Results from adult trials cannot be extrapolated to children because of age-dependent physiologic differences in haemostasis. We studied 22 consecutive paediatric patients who underwent CPB at The Hospital for Sick Children, Toronto. Fibrinogen, factors II, V, VII, VIII, IX, XII, prekallikrein, protein C, protein S, antithrombin (AT), heparin cofactor II, alpha 2-macroglobulin, plasminogen, alpha 2-antiplasmin, tissue plasminogen activator (tPA), plasminogen activator inhibitor, thrombin-AT complexes (TAT), D-dimer, heparin (by both anti-factor Xa assay and protamine titration) and activated clotting time (ACT) were assayed perioperatively. The timing of the sampling was: pre heparin, post heparin, after initiation of CPB, during hypothermia, post hypothermia, post protamine reversal and 24 h post CPB. Plasma concentrations of all haemostatic proteins decreased by an average of 56% immediately following the initiation of CPB due to haemodilution. During CPB, the majority of procoagulants, inhibitors and some components of the fibrinolytic system (plasminogen, alpha 2 AP) remained stable. However, plasma concentrations of TAT and D-dimers increased during CPB showing that significant activation of the coagulation and fibrinolytic systems occurred. Mechanisms responsible for the activation of haemostasis are likely complex. However, low plasma concentrations of heparin (< 2.0 units/ml in 45% of patients) during CPB were likely a major contributing etiology. ACT values showed a poor correlation (r = 0.38) with heparin concentrations likely due to concurrent haemodilution of haemostatic factors, activation of haemostatic system, hypothermia and activation of platelets. In conclusion, CPB in paediatric patients causes global decreases of components of the coagulation and fibrinolytic systems, primarily by haemodilution and secondarily by consumption.


The dose of continuous renal replacement therapy for acute renal failure: a systematic review and meta-analysis.
To conduct a systematic review of the literature to summarize the best available evidence regarding the mortality and morbidity associated with differing dosing regimens of continuous renal replacement therapy (CRRT) for patients with acute renal failure (ARF) in an intensive care unit setting. We searched for randomized controlled trials in electronic databases from January 1990 through November 2009. Eligible trials compared two or more dosing regimens of CRRT in patients with ARF. Two reviewers working independently determined trial eligibility and extracted descriptive, methodological, and outcome data. Random-effects meta-analysis was used to assess relative risks (RR) and weighted mean difference. The I(2)-statistic was used to assess heterogeneity of treatment effect across trials. Seven trials were eligible for meta-analysis. We found no reduction in mortality in patients who received higher doses of CRRT (RR 0.88, 95% CI 0.75-1.03, I(2) = 74%). There was no difference in the requirement of renal replacement therapy at the conclusion of the study period (RR 1.12, 95% CI 0.86-1.46, I(2) = 3%). The overall quality of evidence was downgraded because of imprecision and heterogeneity. Increased dosing of CRRT is not associated with a decrease in mortality of patients with ARF in an intensive care unit setting.


Experimental hypertension in young and adult animals.
The susceptibility of immature and adult animals to various environmental factors often differs because the response of the young organism can only involve those regulatory mechanisms that are available at the particular stage of development. Increased sensitivity to certain (e.g., hypertensive) stimuli may be limited to a relatively short age period that is usually characterized by the maturation of some important physiological functions. High salt intake seems to influence the animals especially during the weaning period and prepuberty, in the course of which profound developmental changes of circulation, electrolyte metabolism, and neurohumoral regulation have been demonstrated. Indeed, salt-dependent forms of experimental hypertension are more severe when they are induced in immature animals. Moreover, substantial differences in hemodynamics, distribution of body fluids, and involvement of pressor and natriuretic agents indicate that the mechanisms of salt hypertension need not be the same in immature and adult animals. For this reason, increased attention should be paid to developmental factors in the study of induced forms of experimental hypertension.


CO Oxidation Promoted by Gold Atoms Loosely Attached in AuFeO3(-) Cluster Anions.
Time-of-flight mass spectrometry experiment shows that upon the interactions with carbon monoxide, the mass-selected AuFeO3(-) oxide cluster anions can evaporate neutral gold atoms in a hexapole collision cell and oxidize CO into CO2 in an ion trap reactor. The computational studies identify that the gold atom is loosely attached in the AuFeO3(-) cluster, and the different reaction channels can be attributed to different cluster velocities. The structure of the AuFeO3(-) cluster is very flexible, and the approach of CO induces significant geometrical and electronic structure changes of AuFeO3(-), which facilitates the exposure of the positively charged gold atom to trap and oxidize CO. The CO oxidation by the AuFeO3(-) cluster follows the Au-assisted Mars-van Krevelen mechanism, in which the direct participation of the surface lattice oxygen (O(2-)) is proposed.


Characterization of early pulmonary hyperproliferation and tumor progression and their inhibition by black tea in a 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone-induced lung tumorigenesis model with A/J mice.
The pathogenesis of pulmonary tumors induced by a tobacco carcinogen, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), and its inhibition by black tea have been characterized. Female A/J mice (6 weeks old) were treated with a single dose of NNK (103 mg/kg of body weight, i.p.) on day 0, and the cell proliferation index was measured by the incorporation of bromodeoxyuridine (BrdUrd) immunohistochemically. The number of BrdUrd-labeled cells increased in the bronchiolar epithelium from day 2 to day 14, with the highest proliferation rate observed on day 5. By day 35, the BrdUrd-labeling index returned to the level of the control group. Further examination of the day 35 samples revealed the presence of foci of hyperproliferative cells in the bronchiolar epithelium, particularly in the bronchiolalveolar regions. These proliferating bronchiolar epithelial cells (Clara cells) may be the initiated sites for pulmonary tumorigenesis. In this short-term model, administration of black tea polyphenols (0.3%) through the drinking water starting 24 h after NNK treatment significantly inhibited NNK-induced early bronchiolar cell proliferation on day 5. In long-term studies, adenomas were observed in 100% (15 of 15) of the mice at week 16, with 7.8 +/- 0.8 tumors per mouse. At week 52, a malignant tumor incidence of 80% (41 of 51 mice) and a malignant tumor multiplicity of 2.39 +/- 0.19 were observed. The growth patterns of the malignant tumors, which included solid, papillary, and mixed types, may be associated with the cellular origin of the tumor. The cell proliferation indices, as measured by proliferating cell nuclear antigen immunohistochemistry, were significantly higher in dysplasia within adenoma than in adenoma, and significantly higher in adenoma at week 52 than in adenoma at week 16. Administration of black tea, starting 16 weeks after a single dose of NNK, inhibited the progression of adenoma to adenocarcinoma as determined by both malignant tumor incidence and multiplicity. The cell proliferation rate in adenomas was also suppressed by black tea treatment. The present work demonstrates the antiproliferative activities of black tea and its polyphenols. Such activities, at the early and late stages of lung tumorigenesis, may be important for the cancer-chemopreventive activities of black tea.


Anticancer activity of an essential oil from Cymbopogon flexuosus.
The essential oil from a lemon grass variety of Cymbopogon flexuosus was studied for its in vitro cytotoxicity against twelve human cancer cell lines. The in vivo anticancer activity of the oil was also studied using both solid and ascitic Ehrlich and Sarcoma-180 tumor models in mice. In addition, the morphological changes in tumor cells were studied to ascertain the mechanism of cell death. The in vitro cytotoxicity studies showed dose-dependent effects against various human cancer cell lines. The IC(50) values of oil ranged from 4.2 to 79 microg/ml depending upon the cell line. In 502713 (colon) and IMR-32 (neuroblastoma) cell lines, the oil showed highest cytotoxicity with IC(50) value of 4.2 and 4.7 microg/ml, respectively. Intra-peritoneal administration of the oil significantly inhibited both ascitic and solid forms of Ehrlich and Sarcoma-180 tumors in a dose-dependent manner. The tumor growth inhibition at 200 mg/kg (i.p.) of the oil observed with both ascitic and solid tumor forms of Ehrlich Ascites carcinoma was 97.34 and 57.83 respectively. In case of Sarcoma-180, the growth inhibition at similar dose of oil was 94.07 and 36.97% in ascitic and solid forms respectively. Morphological studies of the oil treated HL-60 cells revealed loss of surface projections, chromatin condensation and apoptosis. The mitochondria showed apparent loss of cristae in the cells undergoing apoptosis. The morphological studies of Sarcoma-180 solid tumor cells from animals treated with the oil revealed condensation and fragmentation of nuclei typical of apoptosis. Morphological studies of ascites cells from animals treated with the oil too revealed the changes typical of apoptosis. Our results indicate that the oil has a promising anticancer activity and causes loss in tumor cell viability by activating the apoptotic process as identified by electron microscopy.


Catalytic properties of lipoamide dehydrogenase from Mycobacterium smegmatis.
Lipoamide dehydrogenase from Mycobacterium smegmatis was purified to homogeneity over 60-fold. Of 20 amino acid residues identified at the amino terminus of the enzyme, 18 and 17 were identical to the sequences of Mycobacterium leprae and Pseudomonas fluorescens lipoamide dehydrogenases, respectively. The visible spectrum of the isolated enzyme was characteristic of a flavin in apolar environment. Reduction of the enzyme with dithionite results in the appearance of an absorbance shoulder at 530-550 nm, suggesting that reducing equivalents of the two-electron reduced enzyme reside predominantly on the redox-active disulfidedithiol. The kinetic mechanism of the forward (NAD+ reducing) and reverse (NADH oxidizing) reactions proved difficult to study due to severe substrate inhibition by NAD+ and NADH. The rate of lipoamide reduction was found to depend upon the NAD+/NADH ratio, with the reaction being activated at low ratios and inhibited at high ratios. The use of 3-acetylpyridine adenine dinucleotide allowed initial velocity kinetics to be performed and revealed that the kinetic mechanism is ping pong. In addition to catalyzing the reversible oxidation of dihydrolipoamide, the enzyme displayed high oxidase activity (30% of the lipoamide reduction rate), hydrogen and t-butyl peroxide reductase activity (10% of the lipoamide reduction rate), and both naphthoquinone and benzoquinone reduction (approximately 200% of the lipoamide reduction rate). The enzyme failed to catalyze the redox cycling of nitrocompounds, but could anaerobically reduce nitrofurazone. The lipoamide-reducing reaction was reversibly inactivated by sodium arsenite, but no decrease in diaphorase activity was observed under these conditions.


Formation and Infectivity of Oospores of Pseudoperonospora cubensis, the Causal Agent of Downy Mildew in Cucurbits.
The oomycete Pseudoperonospora cubensis attacks members of the Cucurbitaceae, causing severe foliage damage especially to cucumber and melon. Recently, new pathotypes of this oomycete appeared in Israel (2) and Italy (1) and highly aggressive isolates appeared in the United States (3). Since oospores of P. cubensis were rarely seen and sexual propagation by oospores was never reported (4), it is assumed that it propagates clonally by sporangia. Here we report on sexual reproduction of P. cubensis under controlled conditions in the laboratory. We found that field isolates belonging to the old pathotype 3 or to the new pathotype 6 (2) inoculated singly onto detached leaves of cucurbits in growth chambers at 15 or 20°C produced no oospores, even after prolonged incubation periods. However, when sporangia of some paired field isolates were mixed together at a 1:1 ratio, similarly inoculated onto detached leaves, and incubated at 15 or 20°C, numerous oospores (up to ~300/cm2) were formed in the mesophyll within 6 to 11 days, depending on the isolates pair, the host inoculated, and temperature. Oospores were also formed at 12.5°C but not at 25°C. Oospores developed in intact plants when kept at 15 or 20°C under a humidity-saturated atmosphere during disease development. Oospores were round, light brown to brown with an average diameter of ~40 μm. Oospores were produced in Cucumis sativum (cvs. Nadiojni and Dalila) and Cucumis melo (cvs. Ananas-Yokneam and Ein-Dor) but not in Cucurbita pepo (cv. Arlika, Beiruti), C. moschata (cv. Dalorit), or C. maxima (cv. Tripoli). To verify that oospores are infective, cucumber or melon leaves containing oospores were homogenized in water. The homogenate was twice brought to dryness at 25 to 30°C in petri dishes to differentially kill the vegetative structures of the pathogen (sporangia, cystospores, zoospores, and mycelia), resuspended in water, and inoculated onto detached leaves of various cucurbits in growth chambers at 15 or 20°C. Downy mildew lesions carrying sporangia appeared within 7 to 20 days in leaves of Cucumis sativum, Cucumis melo, and C. moschata but not in C. pepo or C. maxima. The recombinant origin of the F1 offspring isolates was confirmed by mefenoxam sensitivity tests, random amplified polymorphic DNA, and simple sequence repeat analyses. F1 progeny isolates of some crosses lost pathogenicity to C. moschata or C. maxima, toward which one of their parents was pathogenic, while others gained pathogenicity to Luffa cylindrica or Citrullus lanatus toward which neither parent was pathogenic. Data confirmed that isolates of P. cubensis can mate to produce oospores, especially under constant humidity conditions; such oospores are infective to cucurbits and F1 progeny isolates show altered sensitivity to fungicides or altered host range relative to their parents. To our knowledge, this is the first report of oospore formation by P. cubensis in the laboratory and on their pathogenicity to cucurbits. Reasons for the parallel appearance of new pathotypes of P. cubensis in Israel in 2002 (2) and Italy in 2003 (1) and the reemergence of highly aggressive isolates of the pathogen in the United States in 2004 (3) are not known. They may be related to oospore production and sexual recombination in P. cubensis. References: (1) C. Cappelli et al. Plant Dis. 87:449, 2003. (2) Y. Cohen et al. Phytoparasitica 31:458, 2003. (3) G. J. Holmes et al. Am. Veg. Grower. February, 14-15, 2006. (4) A. Lebeda and Y. Cohen. Eur. J. Plant Pathol.129:157, 2011.


Dietary fiber, source foods and colorectal cancer risk: the Fukuoka Colorectal Cancer Study.
Despite much evidence from laboratory work, epidemiological evidence remains elusive regarding the role of dietary fiber in colorectal carcinogenesis. We investigated associations of dietary fiber and source foods with colorectal cancer risk in the Fukuoka Colorectal Cancer Study, a community-based case-control study. The study subjects were 816 incident cases of colorectal cancer and 815 community controls. Nutrient and food intakes were estimated on the basis of a computer-assisted interview regarding 148 dietary items. Odds ratios of colorectal cancer according to quintile categories of energy-adjusted intakes of dietary fiber and food groups were obtained with adjustment for non-dietary factors and dietary intakes of calcium and n-3 fatty acids. Total, soluble and insoluble dietary fibers were not measurably associated with overall risk or subsite-specific risk of colorectal cancer. By contrast, rice consumption was associated with a decreased risk of colorectal cancer (trend p = 0.03), particularly of distal colon and rectal cancer (trend p = 0.02), and high intake of non-rice cereals tended to be related to an increased risk of colon cancer (trend p = 0.07). There was no association between vegetable consumption and colorectal cancer, whereas individuals with the lowest intake of fruits tended to have an increased risk of colorectal cancer. The present study did not corroborate a protective association between dietary fiber and colorectal cancer, but suggested a decreased risk of distal colorectal cancer associated with rice consumption.


Activated pancreatic stellate cells can impair pancreatic islet function in mice.
Pancreatic or islet fibrosis is often associated with activated pancreatic stellate cells (PSCs). PSCs are considered not only to promote fibrosis, but also to be associated with glucose intolerance in some diseases. We therefore evaluated morphological and functional relationships between islets and PSCs in the normal mouse pancreas and transplanted islets. Immunohistochemistry was used to map the presence of PSCs in the normal mouse pancreas and islets implanted under the renal capsule. We isolated and cultured mouse PSCs and characterized them morphologically by immunofluorescence staining. Furthermore, we measured their cytokine production and determined their effects on insulin release from simultaneously cultured islets. PSCs were scattered throughout the pancreas, with occasional cells within the islets, particularly in the islet capsule. In islet transplants they were found mainly in the graft periphery. Cultured PSCs became functionally activated and produced several cytokines. Throughout the culture period they linearly increased their production of interleukin-6 and mammalian keratinocyte-derived chemokine. PSC cytokine production was not affected by acute hyperglycemia. Syngeneic islets co-cultured with PSCs for 24-48 h increased their insulin release and lowered their insulin content. However, short-term insulin release in batch-type incubations was unaffected after 48 h of co-culture. Increased islet cell caspase-3 activation and a decreased islet cell replication were consistently observed after co-culture for 2 or 7 days. Activated PSCs may contribute to impaired islet endocrine function seen in exocrine pancreatitis and in islet fibrosis associated with some cases of type 2 diabetes.


The role of ultrasound and lymphoscintigraphy in the assessment of axillary lymph nodes in patients with breast cancer.
Breast cancer is the most common malignancy and the leading cause of death due to cancer in European women. Mammography screening programs aimed to increase the detection of early cancer stages were implemented in numerous European countries. Recent data show a decrease in mortality due to breast cancer in many countries, particularly among young women. At the same time, the number of sentinel node biopsy procedures and breast-conserving surgeries has increased. Intraoperative sentinel lymph node biopsy preceded by lymphoscintigraphy is used in breast cancer patients with no clinical signs of lymph node metastasis. Due to the limited sensitivity and specificity of physical examination in detecting metastatic lesions, developing an appropriate diagnostic algorithm for the preoperative assessment of axillary lymph nodes seems to be a challenge. The importance of ultrasound in patient qualification for sentinel lymph-node biopsy has been discussed in a number of works. Furthermore, different lymphoscintigraphy protocols have been compared in the literature. The usefulness of novel radiopharmaceuticals as well as the methods of image acquisition in sentinel lymph node diagnostics have also been assessed. The aim of this article is to present, basing on current guidelines, literature data as well as our own experience, the diagnostic possibilities of axillary lymph node ultrasound in patient qualification for an appropriate treatment as well as the role of lymphoscintigraphy in sentinel lymph node biopsy.


Dietary intervention restored menses in female athletes with exercise-associated menstrual dysfunction with limited impact on bone and muscle health.
Exercise-related menstrual dysfunction (ExMD) is associated with low energy availability (EA), decreased bone mineral density (BMD), and increased risk of musculoskeletal injury. We investigated whether a 6-month carbohydrate-protein (CHO-PRO) supplement (360 kcal/day, 54 g CHO/day, 20 g PRO/day) intervention would improve energy status and musculoskeletal health and restore menses in female athletes (n = 8) with ExMD. At pre/post-intervention, reproductive and thyroid hormones, bone health (BMD, bone mineral content, bone markers), muscle strength/power and protein metabolism markers, profile of mood state (POMS), and energy intake (EI)/energy expenditure (7 day food/activity records) were measured. Eumenorrheic athlete controls with normal menses (Eumen); n = 10) were measured at baseline. Multiple linear regressions were used to evaluate differences between groups and pre/post-intervention blocking on participants. Improvements in EI (+382 kcal/day; p = 0.12), EA (+417 kcal/day; p = 0.17) and energy balance (EB; +466 kcal/day; p = 0.14) were observed with the intervention but were not statistically significant. ExMD resumed menses (2.6 ± 2.2-months to first menses; 3.5 ± 1.9 cycles); one remaining anovulatory with menses. Female athletes with ExMD for >8 months took longer to resume menses/ovulation and had lower BMD (low spine (ExMD = 3; Eumen = 1); low hip (ExMD = 2)) than those with ExMD for <8 months; for 2 ExMD the intervention improved spinal BMD. POMS fatigue scores were 15% lower in ExMD vs. Eumen (p = 0.17); POMS depression scores improved by 8% in ExMD (p = 0.12). EI, EA, and EB were similar between groups, but the intervention (+360 kcal/day) improved energy status enough to reverse ExMD despite no statistically significant changes in EI. Similar baseline EA and EB between groups suggests that some ExMD athletes are more sensitive to EA and EB fluctuations.


Self-care at school: perceptions of 6-year-old children.
Occupational therapists who work with young children routinely evaluate self-care and consider it an important domain of practice. Little is known about what children perceive is important self-care or what they experience as self-care within a school context. Without knowledge about children's perspectives of self-care, occupational therapists cannot know whether or not they are targeting areas that are central to children's needs. The purpose of this naturalistic study was to explore 6-year-old children's perceptions of self-care in their school day. Participant observation and group interviewing were used to elicit descriptive information from 24 Grade One children, attending an elementary school located in Sydney, Australia. A fishing game, drawing activity, and excerpts from a videotape of their day at school were used as stimuli to explore how the children described and attributed meaning to their self-care occupations. Findings showed that children described self-care at school two ways. First, they named specific self-care tasks that mirrored adult views of self-care and represented culturally shared views of the concept of self-care across ages. Second, children described highly individual views about self-care that were derived from their own experience of doing self-care at school. These views seemed to be based on their personal perceptions of salient factors in operation at the time of self-care performance such as social and physical contexts, perceived skill, and expectations of others. The findings suggest that occupational therapy assessment and intervention for self-care include sensitivity to experiential differences between adult views of self-care and those of children. This sensitivity should include an attempt to understand children's experiences of self-care in specific contexts such as school.


Infantile spasms and encephalopathy without preceding neonatal seizures caused by KCNQ2 R198Q, a gain-of-function variant.
Variants in KCNQ2 encoding for Kv 7.2 neuronal K+ channel subunits lead to a spectrum of neonatal-onset epilepsies, ranging from self-limiting forms to severe epileptic encephalopathy. Most KCNQ2 pathogenic variants cause loss-of-function, whereas few increase channel activity (gain-of-function). We herein provide evidence for a new phenotypic and functional profile in KCNQ2-related epilepsy: infantile spasms without prior neonatal seizures associated with a gain-of-function gene variant. With use of an international registry, we identified four unrelated patients with the same de novo heterozygous KCNQ2 c.593G>A, p.Arg198Gln (R198Q) variant. All were born at term and discharged home without seizures or concern of encephalopathy, but developed infantile spasms with hypsarrhythmia (or modified hypsarrhythmia) between the ages of 4 and 6 months. At last follow-up (ages 3-11 years), all patients were seizure-free and had severe developmental delay. In vitro experiments showed that Kv7.2 R198Q subunits shifted current activation gating to hyperpolarized potentials, indicative of gain-of-function; in neurons, Kv 7.2 and Kv 7.2 R198Q subunits similarly populated the axon initial segment, suggesting that gating changes rather than altered subcellular distribution contribute to disease molecular pathogenesis. We conclude that KCNQ2 R198Q is a model for a new subclass of KCNQ2 variants causing infantile spasms and encephalopathy, without preceding neonatal seizures. A PowerPoint slide summarizing this article is available for download in the Supporting Information section here.


Major variations in Aspergillus fumigatus arising within aspergillomas in chronic pulmonary aspergillosis.
Aspergillomas develop from progressive layers of mycelial growth on the walls of pulmonary cavities over months. Aspergillomas are characteristic of chronic pulmonary aspergillosis and are a risk factor for azole resistance. We investigated genotypic and phenotypic alterations in Aspergillus fumigatus recovered from aspergillomas. Aspergillomas were removed from three patients (two at surgery, one at autopsy) and dissected. Overall 92 colonies of A. fumigatus were isolated. Microsatellite typing was conducted to determine genetic type. Itraconazole, voriconazole and posaconazole susceptibilities were performed. The cyp51A gene was sequenced in 22 isolates. Isolates from Patient 1 (n = 25) were azole susceptible and resistant, although all cyp51A sequences were wild type, the isolates split into two distinct clades. In Patient 2, isolates were less variable (n = 10), all were azole susceptible. In Patient 3 only azole-resistant strains (n = 57) were isolated, with M220K or M220T Cyp51A alterations, and microevolution was indicated. Marked diversity was observed in isolates from these patients; revealing differences in azole susceptibility, mechanism of resistance and genetic type. Importantly, routine sampling from respiratory specimens proved suboptimal in all cases; azole resistance was missed (Patient 1), cultures were negative (Patient 2) and high-level posaconazole resistance was not detected (Patient 3).


Population structure and acquisition of the vanB resistance determinant in German clinical isolates of Enterococcus faecium ST192.
In the context of the global action plan to reduce the dissemination of antibiotic resistances it is of utmost importance to understand the population structure of resistant endemic bacterial lineages and to elucidate how bacteria acquire certain resistance determinants. Vancomycin resistant enterococci represent one such example of a prominent nosocomial pathogen on which nation-wide population analyses on prevalent lineages are scarce and data on how the bacteria acquire resistance, especially of the vanB genotype, are still under debate. With respect to Germany, an increased prevalence of VRE was noted in recent years. Here, invasive infections caused by sequence type ST192 VRE are often associated with the vanB-type resistance determinant. Hence, we analyzed 49 vanB-positive and vanB-negative E. faecium isolates by means of whole genome sequencing. Our studies revealed a distinct population structure and that spread of the Tn1549-vanB-type resistance involves exchange of large chromosomal fragments between vanB-positive and vanB-negative enterococci rather than independent acquisition events. In vitro filter-mating experiments support the hypothesis and suggest the presence of certain target sequences as a limiting factor for dissemination of the vanB element. Thus, the present study provides a better understanding of how enterococci emerge into successful multidrug-resistant nosocomial pathogens.


Phenacyl onium salt photoinitiators: synthesis, photolysis, and applications.
Onium salts, namely sulfonium, phosphonium, ammonium, and pyridinium salts containing phenacyl group are photoinitiators appropriate for the polymerization of monomers such as oxiranes and vinyl ethers, which are not polymerizable by a free-radical mechanism. The initiation is accomplished by direct or indirect (sensitized) photolysis of the salts. Depending on the type of the salt, the direct photoinitiation of cationic polymerization involves reversible or irreversible processes. The photolysis of phenacylsulfonium compounds proceeds by a reversible process, while the other types undergo irreversible photolysis leading to complete fragmentation of the photoinitiator. An additionally useful tool, namely photosensitized generation of initiating species enlarges the versatility of these salts as photoinitiators. Photoinitiated free-radical and zwitterionic polymerizations by using phenacyl-type salts are also addressed. Keto-enol tautomerization of phenacyl pyridinium salts is discussed. Moreover, an interesting application concerning in situ synthesis of clay-poly(methyl methacrylate) nanocomposites with the aid of the phenacyl anilinium salt-based photopolymerization technique is noted.


Local expectation violations result in global activity gain in primary visual cortex.
During natural perception, we often form expectations about upcoming input. These expectations are usually multifaceted - we expect a particular object at a particular location. However, expectations about spatial location and stimulus features have mostly been studied in isolation, and it is unclear whether feature-based expectation can be spatially specific. Interestingly, feature-based attention automatically spreads to unattended locations. It is still an open question whether the neural mechanisms underlying feature-based expectation differ from those underlying feature-based attention. Therefore, establishing whether the effects of feature-based expectation are spatially specific may inform this debate. Here, we investigated this by inducing expectations of a specific stimulus feature at a specific location, and probing the effects on sensory processing across the visual field using fMRI. We found an enhanced sensory response for unexpected stimuli, which was elicited only when there was a violation of expectation at the specific location where participants formed a stimulus expectation. The neural consequences of this expectation violation, however, spread to cortical locations processing the stimulus in the opposite hemifield. This suggests that an expectation violation at one location in the visual world can lead to a spatially non-specific gain increase across the visual field.


Origins of high sequence selectivity: a stopped-flow kinetics study of DNA/RNA hybridization by duplex- and triplex-forming oligonucleotides.
Stopped-flow UV kinetics and thermal denaturation experiments are used to examine the origins of high sequence selectivity and binding affinity of circular triplex-forming oligonucleotides with single-stranded DNA/RNA targets. These 34-nt probes are hybridized to a series of 12-nt target sequences which are fully complementary or which contain a single mismatch. Also studied for comparison are standard 12-nt Watson-Crick DNA or RNA complements. Several novel findings are described: (1) Circular triplex-forming oligomers bind targets with very high thermodynamic selectivity (up to 8-10 kcal/mol against a single-nucleotide mismatch), while linear strands show only 2-3 kcal/mol selectivity. (2) Rates for triplex formation by circular ligands are much greater than other reported triplex formation modes and are nearly the same as for Watson-Crick duplex formation. (3) DNA-DNA and RNA-RNA hybridization rates are similar for both duplex and triplex formation. (4) For both modes of binding, hybridization rates do not vary when a mismatch is introduced into the target, and, therefore, binding selectivity is reflected in large variations in dissociation, rather than association rates. Finally, (5) binding selectivity of circular ligands becomes significantly greater as pH is lowered; results indicate that the high sequence selectivity of the circular DNA ligand is due in large part to the special stability of the protonated C+G-C triad relative to unprotonated mismatched triads. The results are useful in the understanding of properties of nucleic acid complexes in general and give insight into optimum design for synthetic DNA-binding ligands.


Emergence of multi-drug-resistant Acinetobacter anitratus species in neonatal and paediatric intensive care units in a developing country: concern about antimicrobial policies.
Antimicrobial abuse is a serious risk factor for the emergence of multi-drug-resistant (MDR) pathogens. We report on the emergence of multi-drug-resistant Acinetobacter anitratus species over two 8-week periods in 1999 in the neonatal and paediatric intensive care units (NICU and PICU) of King Edward VII Hospital, Durban, South Africa. The source, transmission dynamics, microbiological evaluations, antibiotic utilization patterns and outcome were evaluated. MDR Acinetobacter anitratus was isolated from different body sites in 23 infants less than 2 months of age, 18 in the PICU and five in the NICU. The mortality was 56.5% and two survivors required re-admission. Ten of the 23 cases had entered the ICU with MDR Acinetobacter anitratus. Eleven of the study infants had received broad-spectrum antimicrobial agents before entering the ICU, while all infants with nosocomially acquired MDR Acinetobacter anitratus had received broad-spectrum antimicrobial agents. All the surgical cases with MDR Acinetobacter anitratus died. Microbiological data from both ICUs for 1999 indicated that gram-negative bacteria accounted for two-thirds of isolates, Acinetobacter anitratus and Klebsiella pneumoniae accounting for 33% and 27% of the isolates, respectively. Only 53% and 57% of all Acinetobacter spp isolates were susceptible to piperacillin/tazobactam and carbepenems, respectively. MDR Acinetobacter anitratus is an emerging problem to which antimicrobial abuse contributes.


Rapid and Objective Assessment of Neural Function in Autism Spectrum Disorder Using Transient Visual Evoked Potentials.
There is a critical need to identify biomarkers and objective outcome measures that can be used to understand underlying neural mechanisms in autism spectrum disorder (ASD). Visual evoked potentials (VEPs) offer a noninvasive technique to evaluate the functional integrity of neural mechanisms, specifically visual pathways, while probing for disease pathophysiology. Transient VEPs (tVEPs) were obtained from 96 unmedicated children, including 37 children with ASD, 36 typically developing (TD) children, and 23 unaffected siblings (SIBS). A conventional contrast-reversing checkerboard condition was compared to a novel short-duration condition, which was developed to enable objective data collection from severely affected populations who are often excluded from electroencephalographic (EEG) studies. Children with ASD showed significantly smaller amplitudes compared to TD children at two of the earliest critical VEP components, P60-N75 and N75-P100. SIBS showed intermediate responses relative to ASD and TD groups. There were no group differences in response latency. Frequency band analyses indicated significantly weaker responses for the ASD group in bands encompassing gamma-wave activity. Ninety-two percent of children with ASD were able to complete the short-duration condition compared to 68% for the standard condition. The current study establishes the utility of a short-duration tVEP test for use in children at varying levels of functioning and describes neural abnormalities in children with idiopathic ASD. Implications for excitatory/inhibitory balance as well as the potential application of VEP for use in clinical trials are discussed.


Rapid spread of COVID-19 in New York and the response of the community.
The first COVID-19 patient in New York (NY) was reported on March 1, 2020. Since then NY has become one of the largest epicenters in the world where the disease has been overwhelming the healthcare system. Here I report how rapidly COVID-19 spread, and how the community responded during the first 30 days in NY. Gathering reliable information quickly was important in the evolving situation. Shortage of beds, personal protective equipment, ventilators, and staffing was observed. Reducing the number of infections and increasing the efficiency of medical resource allocation have been two major strategies taken in NY. It is important for Japan to accurately analyze the current situation, refer to answers in other parts of the world, and quickly establish strategy for clear goals that will lead to a "New Normal".


Questionnaire severity measures for depression: a threat to the doctor-patient relationship?
Since 2006 the Quality Outcomes Framework (QOF) has rewarded GPs for carrying out standardised assessments of the severity of symptoms of depression in newly diagnosed patients. To gain understanding of GPs' opinions and perceived impact on practice of the routine introduction of standardised questionnaire measures of severity of depression through the UK general practice contract QOF. Semi-structured qualitative interview study, with purposive sampling and constant comparative analysis. Thirty-four GPs from among 38 study general practices in three sites in England, UK: Southampton, Liverpool, and Norfolk. GPs were interviewed at a time convenient to them by trained interviewers. Interviews were audiorecorded and transcribed verbatim in preparation for thematic analysis, to identify key views. Analysis of the interviews suggested that the use of severity questionnaires posed an intrusion into the consultation. GPs discursively polarised two technologies: formal assessment versus personal enquiry, emphasising the need to ensure the scores are used sensitively and as an aid to clinical judgement rather than as a substitute. Importantly, these challenges implicitly served a function of preserving GPs' identities as professionals with expertise, constructed as integral to the process of diagnosis. GP accounts indicated concern about threats to patient care. Contention between using severity questionnaires and delivering individualised patient care is significantly motivated by GP concerns to preserve professional expertise and identity. It is important to learn from GP concerns to help establish how best to optimise the use of severity questionnaires in depression.


Hepatitis-B vaccination status among dental surgeons in benin city, Nigeria.
The development of success-oriented hepatitis-B vaccine uptake approach among dental surgeons is dependent on the availability of comprehensive baseline data. To determine the hepatitis-B vaccination status among dental surgeons in Benin City. This questionnaire-based cross-sectional study of dental surgeons in Benin City was conducted in May 2011. The questionnaire elicited information on demography, occupational risk rating of contracting hepatitis-B infection, hepatitis-B vaccination status, barriers to uptake of hepatitis vaccine, and suggestions on how to improve hepatitis-B vaccination rates among dental surgeons. Participation rate in the study was 93.3%. More than half (51.4%) of the respondents were 20-30 years old and 52 (74.3%) were males. The occupational risk of contracting hepatitis-B infection among dental surgeons was rated as either high or very high by 51 (72.9%) of the respondents. Amongst the respondents, 14 (20.0%) had received three doses of the hepatitis-B vaccine, 34 (48.6%) either two doses or a single dose, and 22 (31.4%) were not vaccinated. The major barriers reported among the respondents who were not vaccinated were lack of opportunity and the fear of side effects of the vaccines. The suggested ways to increase the vaccination rate among the respondents in descending order include: Making the vaccine available at no cost (51.4%), educating dentists on the merits of vaccination (17.1%), and using the evidence of vaccination as a requirement for annual practicing license renewal (14.3%) and for the employment of dental surgeons (11.4%) and others (2.9%). This study revealed low prevalence of complete hepatitis-B vaccination among the respondents. Improvement in uptake following the respondents' recommendations will serve as a template in developing success-oriented strategies among stakeholders.