Foliar application of biofilm formation-inhibiting compounds enhances control of citrus canker caused by Xanthomonas citri subsp. citri.
Citrus canker caused by the bacterium Xanthomonas citri subsp. citri is an economically important disease of citrus worldwide. Biofilm formation plays an important role in early infection of X. citri subsp. citri on host leaves. In this study, we assessed the hypothesis that small molecules inhibiting biofilm formation reduce X. citri subsp. citri infection and enhance the control of citrus canker disease. D-leucine and 3-indolylacetonitrile (IAN) were found to prevent biofilm formation by X. citri subsp. citri on different abiotic surfaces and host leaves at a concentration lower than the minimum inhibitory concentration (MIC). Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis indicated that IAN repressed expression of chemotaxis/motility-related genes in X. citri subsp. citri. In laboratory experiments, planktonic and biofilm cells of X. citri subsp. citri treated with D-leucine and IAN, either alone or in combination, were more susceptible to copper (CuSO4) than those untreated. In greenhouse assays, D-leucine and IAN applied alone or combined with copper reduced both the number of canker lesions and bacterial populations of X. citri subsp. citri on citrus host leaves. This study provides the basis for the use of foliar-applied biofilm inhibitors for the control of citrus canker alone or combined with copper-based bactericides.


Urticaria.
Urticaria/angioedema is very common and usually not very serious. The main diagnostic task is the history, asking about pharmaceutical agents, foods, focuses of infection, physical agents, and psychogenic factors as well as inhalants, insect bites, internal diseases, immune complex diseases, contactants, and genetic factors. The main therapeutic tool is to eliminate the offending agent. If this cannot be done, therapy should begin with an H1 antihistamine pushed to tolerance or clearing. Life-threatening laryngeal edema and/or anaphylactic shock are extremely rare. Laryngeal edema is usually a component of hereditary angioedema. In such cases, subcutaneous epinephrine is the drug of choice. Laboratory investigation in chronic urticaria should include CBC, erythrocyte sedimentation rate, and a serum multiphasic analysis. A myriad of laboratory tests can be done in chronic urticaria, but some cost-yield effective ones are a test for antinuclear antibodies and x-rays of the sinuses and dentition.


An oral vaccine against NMDAR1 with efficacy in experimental stroke and epilepsy.
The brain is generally considered immunoprivileged, although increasing examples of immunological responses to brain antigens, neuronal expression of major histocompatibility class I genes, and neurological autoimmunity have been recognized. An adeno-associated virus (AAV) vaccine generated autoantibodies that targeted a specific brain protein, the NR1 subunit of the N-methyl-D-aspartate (NMDA) receptor. After peroral administration of the AAV vaccine, transgene expression persisted for at least 5 months and was associated with a robust humoral response in the absence of a significant cell-mediated response. This single-dose vaccine was associated with strong anti-epileptic and neuroprotective activity in rats for both a kainate-induced seizure model and also a middle cerebral artery occlusion stroke model at 1 to 5 months following vaccination. Thus, a vaccination strategy targeting brain proteins is feasible and may have therapeutic potential for neurological disorders.


Acute increase of GABAergic neurotransmission exerts a stimulatory effect on GnRH gene expression in the preoptic/anterior hypothalamic area of ovariectomized, estrogen- and progesterone-treated adult female rats.
Although gamma-aminobutyric acid (GABA) is known to play an important role in the regulation of GnRH release from the hypothalamus, GABAergic action on hypothalamic GnRH gene expression is poorly understood. The present study aims to evaluate the effects of several GABAergic compounds on GnRH mRNA and serum LH levels at the times of LH surge induced by estrogen plus progesterone treatment in long-term ovariectomized adult rats. Animals received either aminooxyacetic acid (AOAA, an inhibitor of GABA catabolism, i.p.), muscimol (GABA-A type agonist, i.c.v.) or baclofen (GABA-B type agonist, i.c.v.) 2 h prior to sacrifice. GnRH mRNA in the preoptic/anterior hypothalamic area and serum LH levels were determined by Northern blot analysis and LH radioimmunoassay, respectively. All of three GABA mimetics blocked the LH surge induced by estrogen plus progesterone in a dose-dependent manner. However, inhibition of GABA catabolism with AOAA in a dose range of 10-100 mg/kg b.w. increased GnRH mRNA level by 30%. Activation of GABA-A receptor with muscimol at a low dose (5 nmol) but not at high doses (10 and 30 nmol) elevated GnRH mRNA levels by 60% over the control value. Activation of GABA-B receptor with baclofen augmented GnRH mRNA levels in a dose-dependent manner. These observations indicate that acute increase of GABAergic neurotransmission may differentially regulate the release and GnRH gene expression depending on its receptor subtypes.


Permeability enhancing lipid-based co-solvent and SEDDS formulations of SQ641, an antimycobacterial agent.
Tuberculosis (TB) is a common and often deadly infectious disease caused by strains of Mycobacteria. Development of new anti-tubercular drugs is essential to control the emergence and severity of multidrug-resistant TB. The objective of this study was to develop an oral preclinical liquid formulation of SQ641 and to determine the permeability across rat intestinal tissue by Ussing chamber. Thermal and chemical characterization of SQ641 was performed by differential scanning calorimetric analysis, thermogravimetric analysis and high performance liquid chromatography. A high throughput solubility screening technique was utilized to determine the solubility of SQ641 in different solvents and co-solvents. Several co-solvent and self-emulsifying drug delivery system (SEDDS) formulations were selected for Ussing chamber permeability studies. Calculated average apparent permeability coefficients of SEDDS formulations of SQ641 (ranging from 0.03 × 10(-6) to 0.33 × 10(-6)) were found to be higher than the permeability coefficients of co-solvent formulations (ranging from 0.00 × 10(-6) to 0.09 × 10(-6)) and those of the neat drug SQ641 in buffer (0.00 × 10(-6)). SEDDS formulations with superior permeability characteristics may provide a useful dosage form for oral intake of anti-tubercular drug SQ641, possibly due to the increase in solubility and immediate dispersion of drug.


Cobalt selenium oxohalides: catalysts for water oxidation.
Two new oxohalides Co4Se3O9Cl2 and Co3Se4O10Cl2 have been synthesized by solid state reactions. They crystallize in the orthorhombic space group Pnma and the monoclinic space group C2/m respectively. The crystal structure of the two compounds are made up of similar building blocks; Co4Se3O9Cl2 is made up of [CoO4Cl2], [CoO5Cl] and [SeO3] polyhedra and Co3Se4O10Cl2 is made up of [CoO4Cl2] and [SeO3] polyhedra. As several Co-containing compounds have proved to be good catalysts for water oxidation, the activities of the two new compounds were compared with the previously found oxohalide Co5Se4O12Cl2 in reference to CoO and CoCl2. The one electron oxidant Ru(bpy)3(3+) was used as oxidizing species in a phosphate buffer and it was found that the activities of the oxohalide species were in between CoO and CoCl2. The roles of Cl(-) and PO4(3-) ions are discussed.


Pig meat quality from entire males.
This paper constitutes an updated review of the production and meat quality aspects of rearing entire male pigs. Since a major obstacle in rearing entire males is the incidence of boar taint, possible methods for detection are also summarised. Safe and fast methods for detection of boar taint would be valuable in avoiding complaints from consumers. Pig meat quality is determined by many aspects, among which odour and taste are the most important attributes. Odour may be negatively affected by the presence of a pheromonal steroid, androstenone, and a fermentation product of l-tryptophan, skatole. Male pigs are surgically castrated in many countries to minimise the risk of accumulation of high levels of androstenone and skatole. Raising entire male pigs is more profitable because they have superior production characteristics and improved meat quality due to leaner carcasses and higher protein content, as compared to castrated pigs. Furthermore, surgical castration is negative from an animal welfare point of view. In most studies, no differences in sensory quality have been found between lean meat from entire male pigs with low levels of androstenone and skatole and pork from castrates and females. The question that remains is: which substances are responsible for boar taint besides androstenone and skatole and whether they need to be considered? The threshold values used for androstenone and skatole might also be too high for highly sensitive persons. Recent research shows that a human odorant receptor, ORD7D4, is involved in sensitivity to androstenone. If the ORD7D4 genotypes of consumer and expert panels are known, this might facilitate consumer studies in the future. There is still a great need for rapid on/at-line detection methods in abattoirs for identifying carcasses with unacceptable levels of boar taint compounds. Several emerging rapid technologies with a potential for boar taint detection have been investigated. They represent various measurement principles such as chemical sensor arrays (electronic noses), mass-spectrometry fingerprinting, ultra-fast gas chromatography, gas-phase spectrometry and biosensors. An industrial detection method should allow 100% correct classification of both acceptable and not-acceptable samples with regard to boar taint sorting criteria. There are, however, still too high a percentage of false negatives ranging from 5% to 20%. In addition, these methods do not yet seem to fulfil the industrial specifications with regard to cost efficiency, simplicity and analysis time. There is still no dedicated measurement technology available for on/at-line detection of boar-tainted carcasses that measures both androstenone and skatole.


Recompleting the Caenorhabditis elegans genome.
Caenorhabditis elegans was the first multicellular eukaryotic genome sequenced to apparent completion. Although this assembly employed a standard C. elegans strain (N2), it used sequence data from several laboratories, with DNA propagated in bacteria and yeast. Thus, the N2 assembly has many differences from any C. elegans available today. To provide a more accurate C. elegans genome, we performed long-read assembly of VC2010, a modern strain derived from N2. Our VC2010 assembly has 99.98% identity to N2 but with an additional 1.8 Mb including tandem repeat expansions and genome duplications. For 116 structural discrepancies between N2 and VC2010, 97 structures matching VC2010 (84%) were also found in two outgroup strains, implying deficiencies in N2. Over 98% of N2 genes encoded unchanged products in VC2010; moreover, we predicted ≥53 new genes in VC2010. The recompleted genome of C. elegans should be a valuable resource for genetics, genomics, and systems biology.


Unraveling the Base Excision Repair Mechanism of Human DNA Glycosylase.
Human DNA glycosylase, hOGG1, is known to perform DNA repair by cleaving oxidized guanine (8OG) from the DNA. Despite numerous experimental and theoretical investigations, the underlying selective molecular mechanism has remained a mystery. Here we present a mechanism that explains how hOGG1's catalytic pocket is able to host an undamaged guanine base, but is not able to cleave it from the DNA. Using linear-scaling quantum mechanics/molecular mechanics (QM/MM) techniques with more than 500 atoms in the QM-region, we have investigated previously proposed mechanisms that all rely on protonating the 8OG nucleobase. We have found that the repair mechanisms propagated in the literature to this date are not capable of differentiating between the G and 8OG nucleobase. Besides this nonselectivity, they also involve reaction barriers that are too high, hence rendering the corresponding reaction intermediates inaccessible. Instead, we present a completely different reaction mechanism, where hOGG1 initially targets the ribose moiety of the substrate and cleaves the glycosidic bond at the very last stage. Our ribose-protonated repair mechanism is not only energetically more preferable, but also explains the selectivity utilized by hOGG1 to block processing a guanine base.


In vitro activity of vancomycin against the spirochete Borrelia burgdorferi.
Borrelia burgdorferi, a spirochete and the causative agent of Lyme disease, has been reported to be susceptible to a variety of antimicrobial agents. In this investigation, the action of vancomycin, a glycopeptide antibiotic not previously known to have activity against spirochetes, against borrelias was examined. The in vitro activity of vancomycin against a variety of strains of B. burgdorferi and one strain of Borrelia hermsii was determined by use of a microdilution MIC method (L.L. Dever, J.H. Jorgensen, and A.G. Barbour, J. Clin. Microbiol. 30:2692-2697, 1992). MICs ranged from 0.5 to 2 micrograms/ml. MICs of the glycopeptides ristocetin and teicoplanin and the lipopeptide daptomycin against strain B31 of B. burgdorferi were all > or = 8 micrograms/ml. Subsurface plating, time-kill studies, synergy studies, and electron microscopy were used to investigate further the activity of vancomycin against B31. The MBC of vancomycin was 2 micrograms/ml. Time-kill curves demonstrated > or = 3-log10-unit (99.9%) killing of the final inoculum after 72 h by vancomycin concentrations twice the MIC. Synergy between vancomycin and penicillin was demonstrated at concentrations one-fourth the MIC of each drug. In electron microscopy, B31 cells exposed to vancomycin showed a disruption of cellular integrity and were indistinguishable from those exposed to penicillin. These studies demonstrate another class of microorganisms susceptible in vitro to vancomycin.


Platelet hyperaggregability in patients with atrial fibrillation. Evidence of a background proinflammatory milieu.
Atrial fibrillation (AF) is a condition where platelet hyperaggregability is commonly present. We examined potential physiological bases for platelet hyperaggregability in a cohort of patients with acute and chronic AF. In particular, we sought to identify the impact of inflammation [myeloperoxidase (MPO) and C-reactive protein (CRP)] and impaired nitric oxide (NO) signaling. Clinical and biochemical determinants of adenosine diphosphate (ADP)-induced platelet aggregation were sought in patients (n = 106) hospitalized with AF via univariate and multivariate analysis. Hyper-responsiveness of platelets to ADP was directly (r = 0.254, p < 0.01) correlated with plasma concentrations of thrombospondin-1 (TSP-1), a matricellular protein that impairs NO responses and contributes to development of oxidative stress. In turn, plasma TSP-1 concentrations were directly correlated with MPO concentrations (r = 0.221, p < 0.05), while MPO concentrations correlated with those of asymmetric dimethylarginine (ADMA, r = 0.220, p < 0.05), and its structural isomer symmetric dimethylarginine (SDMA, r = 0.192, p = 0.05). Multivariate analysis identified TSP-1 (β = 0.276, p < 0.05) concentrations, as well as female sex (β = 0.199, p < 0.05), as direct correlates of platelet aggregability, and SDMA concentrations (β = - 0.292, p < 0.05) as an inverse correlate. We conclude that platelet hyperaggregability, where present in the context of AF, may be engendered by impaired availability of NO, as well as via MPO-related inflammatory activation.


Glutamate receptor genes: susceptibility factors in schizophrenia and depressive disorders?
Schizophrenia, depression, and bipolar disorder are three major neuropsychiatric disorders that are among the leading causes of disability and have enormous economic impacts on our society. Although several neurotransmitter systems have been suggested to play a role in their etiology, we still have not identified any gene or molecular mechanism that might lead to genetic susceptibility for or protection against these neuropsychiatric disorders. The glutamatergic receptor system, and in particular the N-methyl-D-aspartate (NMDA) receptor complex, has long been implicated in their etiology. I review the current molecular evidence that supports a critical role for the glutamatergic receptor system in schizophrenia and the potential involvement of this receptor system in depression and bipolar disorder. It is likely that mutations in glutamate receptor genes might alter the risk of developing one of these disorders. Potential future research directions designed to identify these mutations and to elucidate their effect on mental health will be discussed.


Particleboard made from waste paper treated with maleic anhydride.
This study evaluates the usage of waste newspaper as a supplement raw material for laboratory-made single-layer particle-boards. The boards were made of various ratios (75/25, 50/50, 25/75 and 0/100) of waste paper flakes to wood particles mixtures (wt/wt). The effects of press temperature and maleic anhydride on the properties of the boards were determined. Evaluated properties were modulus of rupture, modulus of elasticity, internal bonding strength, and thickness swelling. Panels made from waste newspaper satisfied the modulus of rupture, modulus of elasticity and internal bonding strength requirements of European Norms standards. Performing a statistical analysis of the results, press temperature and maleic anhydride content were found to be positively effective on the mechanical properties at a 95% confidence level. The results from this study indicated that technically waste paper could be a potential substitute material for particleboard manufacture.


Gaussian-optical approach to stable periodic orbit resonances of partially chaotic dielectric micro-cavities.
The quasi-bound modes localized on stable periodic ray orbits of dielectric micro-cavities are constructed in the short-wavelength limit using the parabolic equation method. These modes are shown to coexist with irregularly spaced ???chaotic??? modes for the generic case. The wavevector quantization rule for the quasi-bound modes is derived and given a simple physical interpretation in terms of Fresnel reflection; quasi-bound modes are explictly constructed and compared to numerical results. The effect of discrete symmetries of the resonator is analyzed and shown to give rise to quasi-degenerate multiplets; the average splitting of these multiplets is calculated by methods from quantum chaos theory.


Effects of unpaired bases on the conformation and stability of three-arm DNA junctions.
Three-arm DNA junctions, in which three double helices intersect at a branch, have unique structure and reactivity of bases at and near the branch. Their solution conformation is asymmetric in the presence of Mg2+, while bases at the branch are sensitive to single-strand-specific agents. Following the surprising report that unpaired bases at the branch stabilize three-arm junctions, we have investigated the geometry and thermodynamics of three-arm junctions containing pendant T and A bases. The results are consistent with additional structure formation in junctions containing up to four pendant bases at the branch: relative to the tight junction, the thermal stability of junctions with two T's or A's at the branch increases; bases near the branch become less reactive to single-strand-reactive probes; and the enthalpy of formation is more negative. The interaction of ethidium observed at the branch in three-arm junctions is enhanced in junctions with unpaired bases at the branch. The geometry of three-arm junctions is perturbed by the presence of pendant bases, as seen by measuring the electrophoretic mobility of junctions to which long duplex arms are appended pairwise.


The Prevalence, Incidence, and Progression of Hand Osteoarthritis in Relation to Body Mass Index, Smoking, and Alcohol Consumption.
To estimate the extent that overweight/obesity, smoking, and alcohol are associated with prevalence and longitudinal changes of radiographic hand osteoarthritis (OA). Participants from the Osteoarthritis Initiative (n = 1232) were included, of whom 994 had 4-year followup data. In analyses on incident hand OA, only persons without hand OA at baseline were included (n = 406). Our exposure variables were overweight/obesity [body mass index (BMI), waist circumference], smoking (current/former, smoking pack-yrs), and alcohol consumption (drinks/week). Using linear and logistic regression analyses, we analyzed possible associations between baseline exposure variables and radiographic hand OA severity, erosive hand OA, incidence of hand OA, and radiographic changes. Analyses were adjusted for age, sex, and education. Neither overweight nor obesity were associated with hand OA. Current smoking was associated with less hand OA in cross-sectional analyses, whereas longitudinal analyses suggested higher odds of incident hand OA in current smokers (OR 2.20, 95% CI 1.02-4.77). Moderate alcohol consumption was associated with higher Kellgren-Lawrence sum score at baseline (1-3 drinks: 1.55, 95% CI 0.43-2.67) and increasing sum score during 4-year followup (4-7 drinks: 0.33, 95% CI 0.01-0.64). Moderate alcohol consumption (1-7 drinks/week) was associated with 2-fold higher odds of erosive hand OA, which was statistically significant. Additional adjustment for BMI gave similar strengths of associations. Overweight/obesity were not associated with hand OA. Contrasting results were observed for smoking and hand OA, suggesting lack of association. Moderate alcohol consumption was associated with hand OA severity, radiographic changes, and erosive hand OA, warranting further investigation.


A Pseudo-Value Approach to Analyze the Semantic Similarity of the Speech of Children With and Without Autism Spectrum Disorder.
Conversational impairments are well known among people with autism spectrum disorder (ASD), but their measurement requires time-consuming manual annotation of language samples. Natural language processing (NLP) has shown promise in identifying semantic difficulties when compared to clinician-annotated reference transcripts. Our goal was to develop a novel measure of lexico-semantic similarity - based on recent work in natural language processing (NLP) and recent applications of pseudo-value analysis - which could be applied to transcripts of children's conversational language, without recourse to some ground-truth reference document. We hypothesized that: (a) semantic coherence, as measured by this method, would discriminate between children with and without ASD and (b) more variability would be found in the group with ASD. We used data from 70 4- to 8-year-old males with ASD (N = 38) or typically developing (TD; N = 32) enrolled in a language study. Participants were administered a battery of standardized diagnostic tests, including the Autism Diagnostic Observation Schedule (ADOS). ADOS was recorded and transcribed, and we analyzed children's language output during the conversation/interview ADOS tasks. Transcripts were converted to vectors via a word2vec model trained on the Google News Corpus. Pairwise similarity across all subjects and a sample grand mean were calculated. Using a leave-one-out algorithm, a pseudo-value, detailed below, representing each subject's contribution to the grand mean was generated. Means of pseudo-values were compared between the two groups. Analyses were co-varied for nonverbal IQ, mean length of utterance, and number of distinct word roots (NDR). Statistically significant differences were observed in means of pseudo-values between TD and ASD groups (p = 0.007). TD subjects had higher pseudo-value scores suggesting that similarity scores of TD subjects were more similar to the overall group mean. Variance of pseudo-values was greater in the ASD group. Nonverbal IQ, mean length of utterance, or NDR did not account for between group differences. The findings suggest that our pseudo-value-based method can be effectively used to identify specific semantic difficulties that characterize children with ASD without requiring a reference transcript.


Cholesterol uptake disruption, in association with chemotherapy, is a promising combined metabolic therapy for pancreatic adenocarcinoma.
The malignant progression of pancreatic ductal adenocarcinoma (PDAC) is accompanied by a profound desmoplasia, which forces proliferating tumor cells to metabolically adapt to this new microenvironment. We established the PDAC metabolic signature to highlight the main activated tumor metabolic pathways. Comparative transcriptomic analysis identified lipid-related metabolic pathways as being the most highly enriched in PDAC, compared with a normal pancreas. Our study revealed that lipoprotein metabolic processes, in particular cholesterol uptake, are drastically activated in the tumor. This process results in an increase in the amount of cholesterol and an overexpression of the low-density lipoprotein receptor (LDLR) in pancreatic tumor cells. These findings identify LDLR as a novel metabolic target to limit PDAC progression. Here, we demonstrate that shRNA silencing of LDLR, in pancreatic tumor cells, profoundly reduces uptake of cholesterol and alters its distribution, decreases tumor cell proliferation, and limits activation of ERK1/2 survival pathway. Moreover, blocking cholesterol uptake sensitizes cells to chemotherapeutic drugs and potentiates the effect of chemotherapy on PDAC regression. Clinically, high PDAC Ldlr expression is not restricted to a specific tumor stage but is correlated to a higher risk of disease recurrence. This study provides a precise overview of lipid metabolic pathways that are disturbed in PDAC. We also highlight the high dependence of pancreatic cancer cells upon cholesterol uptake, and identify LDLR as a promising metabolic target for combined therapy, to limit PDAC progression and disease patient relapse.


Paraneoplastic encephalitis, psychiatric symptoms, and hypoventilation in ovarian teratoma.
We report four young women who developed acute psychiatric symptoms, seizures, memory deficits, decreased level of consciousness, and central hypoventilation associated with ovarian teratoma (OT) and cerebrospinal fluid (CSF) inflammatory abnormalities. Three patients recovered with treatment of the tumor or immunosuppression and one died of the disorder. Five other OT patients with a similar syndrome and response to treatment have been reported. Our patients' serum or CSF showed immunolabeling of antigens that were expressed at the cytoplasmic membrane of hippocampal neurons and processes and readily accessed by antibodies in live neurons. Immunoprobing of a hippocampal-expression library resulted in the isolation of EFA6A, a protein that interacts with a member of the two-pore-domain potassium channel family and is involved in the regulation of the dendritic development of hippocampal neurons. EFA6A-purified antibodies reproduced the hippocampal immunolabeling of all patients' antibodies and colocalized with them at the plasma membrane. These findings indicate that in a young woman with acute psychiatric symptoms, seizures, and central hypoventilation, a paraneoplastic immune-mediated syndrome should be considered. Recognition of this disorder is important because despite the severity of the symptoms, patients usually recover. The location and function of the isolated antigen suggest that the disorder is directly mediated by antibodies.


Engineering antenna radiation patterns via quasi-conformal mappings.
We use a combination of conformal and quasi-conformal mappings to engineer isotropic electromagnetic devices that modify the omnidirectional radiation pattern of a point source. For TE waves, the designed devices are also non-magnetic. The flexibility offered by the proposed method is much higher than that achieved with conformal mappings. As a result, it is shown that complex radiation patterns can be achieved, which can combine high directivity in a desired number of arbitrary directions and isotropic radiation in other specified angular ranges. In addition, this technique enables us to control the power radiated in each direction to a certain extent. The obtained results are valid for any part of the spectrum. The potential of this method is illustrated with some examples. Finally, we study the frequency dependence of the considered devices and propose a practical dielectric implementation.


Vaccine-Induced Protection from Homologous Tier 2 SHIV Challenge in Nonhuman Primates Depends on Serum-Neutralizing Antibody Titers.
Passive administration of HIV neutralizing antibodies (nAbs) can protect macaques from hard-to-neutralize (tier 2) chimeric simian-human immunodeficiency virus (SHIV) challenge. However, conditions for nAb-mediated protection after vaccination have not been established. Here, we selected groups of 6 rhesus macaques with either high or low serum nAb titers from a total of 78 animals immunized with recombinant native-like (SOSIP) Env trimers. Repeat intrarectal challenge with homologous tier 2 SHIVBG505 led to rapid infection in unimmunized and low-titer animals. High-titer animals, however, demonstrated protection that was gradually lost as nAb titers waned over time. An autologous serum ID50 nAb titer of ∼1:500 afforded more than 90% protection from medium-dose SHIV infection. In contrast, antibody-dependent cellular cytotoxicity and T cell activity did not correlate with protection. Therefore, Env protein-based vaccination strategies can protect against hard-to-neutralize SHIV challenge in rhesus macaques by inducing tier 2 nAbs, provided appropriate neutralizing titers can be reached and maintained.


High-altitude ancestry and hypoxia acclimation have distinct effects on exercise capacity and muscle phenotype in deer mice.
The hypoxic and cold environment at high altitudes requires that small mammals sustain high rates of O2 transport for exercise and thermogenesis while facing a diminished O2 availability. We used laboratory-born and -raised deer mice (Peromyscus maniculatus) from highland and lowland populations to determine the interactive effects of ancestry and hypoxia acclimation on exercise performance. Maximal O₂consumption (V̇o(2max)) during exercise in hypoxia increased after hypoxia acclimation (equivalent to the hypoxia at ∼4,300 m elevation for 6-8 wk) and was consistently greater in highlanders than in lowlanders. V̇o(2max) during exercise in normoxia was not affected by ancestry or acclimation. Highlanders also had consistently greater capillarity, oxidative fiber density, and maximal activities of oxidative enzymes (cytochrome c oxidase and citrate synthase) in the gastrocnemius muscle, lower lactate dehydrogenase activity in the gastrocnemius, and greater cytochrome c oxidase activity in the diaphragm. Hypoxia acclimation did not affect any of these muscle traits. The unique gastrocnemius phenotype of highlanders was associated with higher mRNA and protein abundances of peroxisome proliferator-activated receptor γ (PPARγ). Vascular endothelial growth factor (VEGFA) transcript abundance was lower in highlanders, and hypoxia acclimation reduced the expression of numerous genes that regulate angiogenesis and energy metabolism, in contrast to the observed population differences in muscle phenotype. Lowlanders exhibited greater increases in blood hemoglobin content, hematocrit, and wet lung mass (but not dry lung mass) than highlanders after hypoxia acclimation. Genotypic adaptation to high altitude, therefore, improves exercise performance in hypoxia by mechanisms that are at least partially distinct from those underlying hypoxia acclimation.


Effective treatment of Streptococcus uberis clinical mastitis to minimize the use of antibiotics.
Antibiotic regimens (intramammary antibiotic, penicillin-based parenteral treatment) and intramuscular oxytocin were tested for effectiveness against experimental infection by Streptococcus uberis with the following results from 54 animals: a) no treatment led to deterioration of infected quarters, requiring intervention within 48 h for cow health; b) aggressive intramammary antibiotic at every milking achieved 70% clinical cure in 3 d and 100% cure within 6 d; overall bacteriological cure was 80%; c) parenteral treatment alone used about 14 times as much antibiotic with 18% clinical cure in 3 d and 91% within 6 d; overall bacteriological cure was 80%; d) combination of aggressive intramammary and parenteral treatments achieved 61% clinical cure in 3 d and 100% within 6 d; overall bacteriological cure was 72%; e) intramammary antibiotic at labeled rates (1x for 3 d) achieved 27% clinical cure in 3 d but 91% within 6 d of treatment; overall bacteriological cure was 64%; f) use of oxytocin alone for 3 d failed to achieve clinical improvement with an increase in the severity of mastitis; g) combining oxytocin with labeled use of intramammary antibiotic (1x for 3 d) was unsuccessful: 0% clinical cures in 3 d, 10% in 6 d; significantly poorer than intramammary antibiotic alone. Extended treatment periods with parenteral or intramammary antibiotics resulted in positive inhibitory tests for milk from individual quarters up to 8 d after treatment. Aggressive intramammary antibiotic was the most effective treatment for fastest cure clinically and bacteriologically using least antibiotic.


Ethical Arguments Concerning the Use of Alzheimer's Disease Biomarkers in Individuals with No or Mild Cognitive Impairment: A Systematic Review and Framework for Discussion.
The shift to defining Alzheimer's disease (AD) as a biological continuum, which is characterized by the presence of biomarkers instead of clinical symptoms, has sparked a widespread debate. Insight into the given arguments and their underlying moral values is crucial to ensure well-considered and appropriate AD biomarker testing in the future. To critically review the arguments in favor of or against AD biomarker testing in people with no or mild cognitive impairment and to explicate their underlying moral values. Seven databases were systematically searched for publications mentioning arguments of interest. Arguments are identified using qualitative data-analysis and evaluated within an ethical framework. Our search yielded 3,657 articles of which 34 met the inclusion criteria. We discuss the clusters of arguments separate from their evaluation and the assessment of the debate as a whole. The right to know, which derives from the moral value of respect for autonomy, is a central argument in favor of biomarker testing. On the other hand, fear of the disease and lack of a disease-modifying treatment may result in a negative balance of good over inflicted harms, which argues against its use. Critical evaluation and weighing of the given arguments in a specific context, within an ethical framework, demonstrates the necessity to differentiate between what we hope or expect from research and where we currently stand. While AD biomarkers may have an indispensable value for research, the current advantage for clinical practice appears limited.


Orthogonal reactivity of metal and multimetal nanostructures for selective, stepwise, and spatially-controlled solid-state modification.
Chemists rely on a toolbox of robust chemical transformations for selectively modifying molecules with spatial and functional precision to make them more complex in a controllable and predictable manner. This manuscript describes proof-of-principle experiments for a conceptually analogous strategy involving the selective, stepwise, and spatially controlled modification of inorganic nanostructures. The key concept is orthogonal reactivity: one component of a multicomponent system reacts with a particular reagent under a specific set of conditions while the others do not, even though they are all present together in the same reaction vessel. Using the chemical conversion of metal nanoparticles into intermetallic, sulfide, and phosphide nanoparticles as representative examples, the concept of orthogonal reactivity is defined and demonstrated for a variety of two- and three-component nanoscale systems. First, solution-phase reactivity data are presented and collectively analyzed for the reaction of metal nanoparticles (Ni, Cu, Rh, Pd, Ag, Pt, Au, Sn) with several metal salt and elemental reagents (Bi, Pb, Sb, Sn, S). From these data, several two- and three-component orthogonal systems are identified. Finally, these results are applied to the spatially selective chemical modification of lithographically patterned surfaces and striped template-grown metal nanowires.


Nonocclusive mesenteric ischemia in a patient on maintenance hemodialysis.
Nonocclusive mesenteric ischemia (NOMI) is known to occupy about 25% to 60% of intestinal infarction. NOMI has been reported to be responsible for 9% of the deaths in the dialysis population and the postulated causes of NOMI include intradialytic hypotension, atherosclerosis and medications, such as diuretics, digitalis and vasopressors. Clinical manifestations, such as fever, diarrhea and leukocytosis, are nonspecific, which makes early diagnosis of NOMI very difficult. A 66-year-old woman on maintenance hemodialysis for 5 years was admitted with syncope, abdominal pain and chilly sensation. Since 7 days prior to admission, blood pressure on the supine position during hemodialysis had frequently fallen to 80/50 mmHg. Four days later, she complained of progressive abdominal pain. Rebound tenderness and leukocytosis (WBC 13900/mm3) with left shift were noted. Stool examination was positive for occult blood. Abdominal CT scan showed a distended gall bladder with sludge. Under the impression of acalculous cholecystitis, she was operated on. Surgical and pathologic findings of colon colon were compatible with NOMI. Because of recurrent intradialytic hypotension, we started midodrine 2.5 mg just before hemodialysis and increased the dose up to 7.5 mg. After midodrine therapy, blood pressure during dialysis became stable and the symptoms associated with hypotension did not recur. As NOMI may occur within several hours or days after an intradialytic hypotensive episode, abdominal pain should be carefully observed and NOMI should be considered as a differential diagnosis. In addition, we suggest that midodrine be considered to prevent intradialytic hypotensive episodes.


EH-myomesin splice isoform is a novel marker for dilated cardiomyopathy.
The M-band is the prominent cytoskeletal structure that cross-links the myosin and titin filaments in the middle of the sarcomere. To investigate M-band alterations in heart disease, we analyzed the expression of its main components, proteins of the myomesin family, in mouse and human cardiomyopathy. Cardiac function was assessed by echocardiography and compared to the expression pattern of myomesins evaluated with RT-PCR, Western blot, and immunofluorescent analysis. Disease progression in transgenic mouse models for dilated cardiomyopathy (DCM) was accompanied by specific M-band alterations. The dominant splice isoform in the embryonic heart, EH-myomesin, was strongly up-regulated in the failing heart and correlated with a decrease in cardiac function (R = -0.86). In addition, we have analyzed the expressions of myomesins in human myocardial biopsies (N = 40) obtained from DCM patients, DCM patients supported by a left ventricular assist device (LVAD), hypertrophic cardiomyopathy (HCM) patients and controls. Quantitative RT-PCR revealed that the EH-myomesin isoform was up-regulated 41-fold (P < 0.001) in the DCM patients compared to control patients. In DCM hearts supported by a LVAD and HCM hearts, the EH-myomesin expression was comparable to controls. Immunofluorescent analyses indicate that EH-myomesin was enhanced in a cell-specific manner, leading to a higher heterogeneity of the myocytes' cytoskeleton through the myocardial wall. We suggest that the up-regulation of EH-myomesin denotes an adaptive remodeling of the sarcomere cytoskeleton in the dilated heart and might serve as a marker for DCM in mouse and human myocardium.


A randomised, controlled trial of the effects of a mobile telehealth intervention on clinical and patient-reported outcomes in people with poorly controlled diabetes.
Objective The objective of this research is to determine the effects of mobile telehealth (MTH) on glycosylated haemoglobin (HbA1c) and other clinical and patient-reported outcomes in insulin-requiring people with diabetes. Methods A nine-month randomised, controlled trial compared standard care to standard care supplemented with MTH (self-monitoring, mobile-phone data transmissions, graphical and nurse-initiated feedback, and educational calls). Clinical (HbA1c, blood pressure, daily insulin dose, diabetes outpatient appointments (DOAs)) and questionnaire data (health-related quality of life, depression, anxiety) were collected. Mean group changes over time were compared using hierarchical linear models and Mann-Whitney tests. Results Eighty-one participants with a baseline HbA1c of 8.98% ± 1.82 were randomised to the intervention ( n = 45) and standard care ( n = 36). The Group by Time effect revealed MTH did not significantly influence HbA1c ( p = 0.228), but p values were borderline significant for blood pressure ( p = 0.054) and mental-health related quality of life ( p = 0.057). Examination of effect sizes and 95% confidence intervals for mean group differences at nine months supported the existence of a protective effect of MTH on mental health-related quality of life as well as depression. None of the other measured outcomes were found to be affected by the MTH intervention. Conclusions Findings from this study must be interpreted with caution given the small sample size, but they do not support the widespread adoption of MTH to achieve clinically significant changes in HbA1c. MTH may, however, have positive effects on blood pressure and protective effects on some aspects of mental health.


Collagen formation observed from healing calvarial defects with principal component analysis of Raman scattering.
Bone healing is a complex process involving molecular changes. Bone matrix consists of collagen proteins that serve as the framework and minerals, calcium and phosphate, are deposited into the matrix accordingly. Raman spectroscopy is a promising technique to study bone mineral and matrix environments simultaneously. We studied the bone composition using 785 nm excitation during healing of subcritical calvarial defects without disrupting the fracture. Calvarial defects (in vivo) were created using a 1 mm burr drill on the parietal bones of Sprague-Dawley rats (n = 12). After 7 days, subjects were sacrificed and an additional defect (control) was created. Principal component analysis was utilized for the analysis of Raman spectra and helped in classifying normal and healing bone. Principal component 1 (PC1) shows that the major variation between in vivo and control defects and normal bone surface is at 958 cm-1 (ν1 phosphate band). PC2 shows a major variation at 1448 cm-1 (CH2 deformation). PC2 score distinguishes in vivo defects from normal surface and control defects. The decrease in crystallinity and mineral to matrix ratio at the healing site as revealed by Raman confirms the new bone formation. Scanning electron and optical microscopy show the formation of newly generated matrix by means of bony bridges of collagens. The surface roughness increases by 23% from control to in vivo defects, as revealed by optical profiler. Histology shows the decreased depth of in vivo defects and new blood vessels formation. Overall, the new collagen formation shows the scaffolding of the bone is growing during healing.


Comparative connectomics of the primate social brain.
Social interaction is thought to provide a selection pressure for human intelligence, yet little is known about its neurobiological basis and evolution throughout the primate lineage. Recent advances in neuroimaging have enabled whole brain investigation of brain structure, function, and connectivity in humans and non-human primates (NHPs), leading to a nascent field of comparative connectomics. However, linking social behavior to brain organization across the primates remains challenging. Here, we review the current understanding of the macroscale neural mechanisms of social behaviors from the viewpoint of system neuroscience. We first demonstrate an association between the number of cortical neurons and the size of social groups across primates, suggesting a link between neural information-processing capacity and social capabilities. Moreover, by capitalizing on recent advances in species-harmonized functional MRI, we demonstrate that portions of the mirror neuron system and default-mode networks, which are thought to be important for representation of the other's actions and sense of self, respectively, exhibit similarities in functional organization in macaque monkeys and humans, suggesting possible homologies. With respect to these two networks, we describe recent developments in the neurobiology of social perception, joint attention, personality and social complexity. Together, the Human Connectome Project (HCP)-style comparative neuroimaging, hyperscanning, behavioral, and other multi-modal investigations are expected to yield important insights into the evolutionary foundations of human social behavior.


The identification of clinical and professional leadership activities of advanced practitioners: findings from the Specialist Clinical and Advanced Practitioner Evaluation study in Ireland.
To report a case study that identifies how leadership is enacted by advanced practitioners in nursing and midwifery and differentiates between clinical and professional leadership in advanced practice. A national evaluation of advanced practitioners in nursing and midwifery was commissioned to inform policy makers' decision-making about future developments in Ireland. Leadership is a key aspect of the advanced practitioner role. However, leadership in the advanced practice context is under researched and lacks empirical evidence on how advanced practitioners enact their clinical and professional leadership function in practice. A multiple case study methodology was used to identify the leadership activities of advanced practitioners. Purposive sampling of advanced practitioners in nursing and midwifery across various specialities and health regions in Ireland was employed. Data sources included non-participant observation of advanced practitioners, interviews with key stakeholders, and documentary evidence including audits, policies, and guidelines. Collectively these data were used to identify the leadership activities of advanced practitioners in practice. The study was conducted between December 2008-December 2010. Seven key activities that indicated clinical leadership and three key activities for professional leadership at advanced practice level were identified. The leadership activities identified provide greater clarity in distinguishing between professional and clinical leadership and in specifying the nature and scope of leadership activities.


Trypanosoma cruzi infection in dogs and cats and household seroreactivity to T. cruzi in a rural community in northeast Brazil.
The prevalence of Trypanosoma cruzi parasitemia as determined by xenodiagnosis on domestic dogs and cats was correlated with household rates of seroreactivity to T. cruzi and household Panstrongylus megistus infestation in a rural area in northeast Brazil where P. megistus was the only domiciliary triatomine vector. T. cruzi infection was present in about 18% of domestic dogs and cats. Two-thirds of seroreactive children below age 10 resided in houses with T. cruzi-infected animals. In houses with a T. cruzi-infected dog or cat, as well as at least one infected P. megistus, the household rate of seroreactivity to T. cruzi was five times greater than in houses with non-infected domestic animals and no infected triatomine vectors. Domestic dogs and cats are important reservoirs of T. cruzi in an endemic area where P. megistus is the only domiciliary triatomine vector.


Effect of cognitive remediation on gait in sedentary seniors.
Attention and executive functions show strong associations with slow gait and falls in seniors and have been shown to be amenable to cognitive remediation. However, cognitive remediation as a strategy to improve mobility has not been investigated. Using a randomized single-blind control design, 24 sedentary older adults (exercise less than or equal to once weekly and gait velocity <1 m/s) were randomly assigned to an 8-week computerized cognitive remediation program or wait-list. Primary outcome was change in gait velocity during normal pace and "walking while talking" conditions. We also compared the proportion of improvers (velocity change ≥4 cm/s) in each group. The 10 participants who completed cognitive remediation improved gait velocity from baseline during normal walking (68.2 ± 20.0 vs 76.5 ± 17.9 cm/s, p = .05) and walking while talking (36.7 ± 13.5 vs 56.7 ± 20.4 cm/s, p = .002). The 10 intervention participants improved gait velocity over the 8-week intervention both during normal walking (change: 8.2 ± 11.4-1.3 ± 6.8 cm/s, p = .10) and walking while talking (change: 19.9 ± 14.9-2.5 ± 20.1 cm/s, p = .05) compared with the 10 control participants. Six intervention participants were improvers on normal pace walking compared with three controls (odds ratio = 3.0, 95% confidence interval = 0.5-19.6). All 10 intervention participants improved on walking while talking compared with 3 controls (odds ratio = 3.5, 95% confidence interval = 1.5-8.0). The findings of this pilot trial are promising and suggest that cognitive remediation may improve mobility in sedentary seniors. This approach should be validated in larger scale trials.


Personal view: adequate relief as a primary endpoint in irritable bowel syndrome.
The success of a clinical trial is judged by achieving statistical and clinical significance on the primary endpoint. This is particularly relevant in those trials that are initiated to achieve regulatory approval of a new therapeutic agent. Selection of an endpoint for which statistical significance will be too difficult to achieve can result in clinical trial data that fails to meet regulatory requirements for product approval. Over the past decade, significant progress has been made in development and refinement of the study design for evaluating new therapeutic agents targeted for the treatment of irritable bowel syndrome. One aspect of trial design that has been advanced is recognition of the endpoint of 'Adequate Relief.' Adequate relief has been used as a primary endpoint in treatment trials with alosetron, cilansetron and dextofisopam. With each of these agents, statistically significant benefit was seen when compared to placebo. As an endpoint, adequate relief is responsive, reproducible and moves in the same direction as other meaningful measures, and, therefore, displays properties of a validated endpoint. Adequate relief should be considered an acceptable primary endpoint by regulatory agencies for use in clinical trials of novel therapeutic agents for irritable bowel syndrome.


Changing Trends in Brain Imaging Technique for Pediatric Patients with Ventriculoperitoneal Shunts.
Children with ventriculoperitoneal shunts (VPS) undergoing brain computed tomography (CT) for shunt malfunction evaluation are at risk for later malignancy due to radiation exposure. We aimed to determine if and how hospitals have adopted radiation-avoiding magnetic resonance imaging (MRI) techniques. We performed a secondary analysis of the Pediatric Health Information System (PHIS) database. Children with VPS presenting to acute wards at 31 PHIS hospitals between January 1, 2007 and January 2, 2015 and receiving noncontrast neuroimaging on day of service 0/1 were included. Outcome measures were (1) incidence of MRI over time and (2) comparison of demographic characteristics between hospitals with MRI representing higher versus lower proportions (>15% or <15%) of total brain imaging. MRIs increased by 18.1% from 2007 to 2015. Hospitals were assigned to high-use (n = 12) or minimal-use (n = 19) MRI groups based on year 2014/2015 MRI percentages. The only identified difference was an older mean age in the high-use group (8.1 vs. 7.5 years; p = 0.03). MRI is increasingly used to evaluate patients with VPS. Hospitals with more MRI use had older patients and no increase in cost or length of stay. Initiating local quality improvement projects may help identify barriers to MRI uptake and increase use.


Genome-wide identification, characterization, and expression analysis of lineage-specific genes within zebrafish.
The genomic basis of teleost phenotypic complexity remains obscure, despite increasing availability of genome and transcriptome sequence data. Fish-specific genome duplication cannot provide sufficient explanation for the morphological complexity of teleosts, considering the relatively large number of extinct basal ray-finned fishes. In this study, we performed comparative genomic analysis to discover the Conserved Teleost-Specific Genes (CTSGs) and orphan genes within zebrafish and found that these two sets of lineage-specific genes may have played important roles during zebrafish embryogenesis. Lineage-specific genes within zebrafish share many of the characteristics of their counterparts in other species: shorter length, fewer exon numbers, higher GC content, and fewer of them have transcript support. Chromosomal location analysis indicated that neither the CTSGs nor the orphan genes were distributed evenly in the chromosomes of zebrafish. The significant enrichment of immunity proteins in CTSGs annotated by gene ontology (GO) or predicted ab initio may imply that defense against pathogens may be an important reason for the diversification of teleosts. The evolutionary origin of the lineage-specific genes was determined and a very high percentage of lineage-specific genes were generated via gene duplications. The temporal and spatial expression profile of lineage-specific genes obtained by expressed sequence tags (EST) and RNA-seq data revealed two novel properties: in addition to being highly tissue-preferred expression, lineage-specific genes are also highly temporally restricted, namely they are expressed in narrower time windows than evolutionarily conserved genes and are specifically enriched in later-stage embryos and early larval stages. Our study provides the first systematic identification of two different sets of lineage-specific genes within zebrafish and provides valuable information leading towards a better understanding of the molecular mechanisms of the genomic basis of teleost phenotypic complexity for future studies.


Causes and circumstances of neonatal deaths in 108 consecutive cases over a 10-year period at the Children's Hospital of Lucerne, Switzerland.
Neonatal deaths still represent the largest percentage of overall childhood mortality. Many deaths of neonates are preceded by end-of-life decisions; however, decision-making practices have been reported to vary widely from country to country. To analyze principal causes and circumstances of all consecutive neonatal deaths at our institution over a 10-year period. All neonates who had died either in the delivery room (DR) or the neonatal intensive care unit (NICU) between January 1, 1997 and December 31, 2006 were identified. Demographic information, principal causes and circumstances of death were abstracted from the individual medical records. There were approximately 72,000 live births in the catchment area of our center with 15,150 deliveries occurring at the Women's Hospital of Lucerne. Of the 108 deaths identified, 29 occurred in the DR (DR mortality rate 0.2%) and 79 in the NICU (NICU mortality rate 2.3%). The majority of DR deaths occurred in the setting of primary nonintervention and were related to extreme prematurity (n = 20), lethal congenital malformations (n = 6) and trisomy 13 (n = 2). One patient with severe perinatal asphyxia died despite full resuscitative efforts. In the NICU, overall mortality rate was inversely related to gestational age (GA). Cardiovascular and respiratory system failures were the predominant causes of death in premature infants with a GA <32 weeks, whereas CNS catastrophes accounted for the majority of deaths in the more mature NICU population. End-of-life decisions were common with less than 10% of deaths occurring despite maximal intensive care. In our perinatal center, primary nonintervention and redirection of care are the most common circumstances of death in neonates. This reflects our belief that, apart from futility, quality-of-life considerations are an important part of decision making in neonatology.


Analysis of thrombosis on postoperative day 5 or later after microvascular reconstruction for head and neck cancers.
Because of the low incidence of late thrombosis in free flaps used for head and neck reconstruction, the risk factors, prognosis, and the optimal method of treatment are unclear. The timing of thrombosis, types of flaps, occluded vessels, causative factors, previous irradiation, and salvage rates were reviewed and compared between 79 patients who had thrombosis on postoperative day 4 or earlier (early-thrombosis group) and 24 patients who had thrombosis on postoperative day 5 or later (late-thrombosis group). The main causative factor for thrombosis in the late-thrombosis group was wound infection (54%), whereas wound infection was present in only 1% of cases of thrombosis in the early-thrombosis group. None of the flaps could be salvaged in the late-thrombosis group. Poor salvage rate in the late-thrombosis group is the most serious problem. Prevention, early detection, and appropriate management of wound infection are essential for avoiding late thrombosis.


Oncogenes induce genotoxic stress by mitotic processing of unusual replication intermediates.
Oncogene-induced DNA replication stress activates the DNA damage response (DDR), a crucial anticancer barrier. DDR inactivation in these conditions promotes genome instability and tumor progression, but the underlying molecular mechanisms are elusive. We found that overexpression of both Cyclin E and Cdc25A rapidly slowed down replication forks and induced fork reversal, suggestive of increased topological stress. Surprisingly, these phenotypes, per se, are neither associated with chromosomal breakage nor with significant DDR activation. Oncogene-induced DNA breakage and DDR activation instead occurred upon persistent G2/M arrest or, in a checkpoint-defective context, upon premature CDK1 activation. Depletion of MUS81, a cell cycle-regulated nuclease, markedly limited chromosomal breakage and led to further accumulation of reversed forks. We propose that nucleolytic processing of unusual replication intermediates mediates oncogene-induced genotoxicity and that limiting such processing to mitosis is a central anti-tumorigenic function of the DNA damage checkpoints.


Alternative lengthening of telomeres phenotype in malignant vascular tumors is highly associated with loss of ATRX expression and is frequently observed in hepatic angiosarcomas.
Alternative lengthening of telomeres (ALT) is a mechanism using homologous recombination to maintain telomere length and sustain limitless replicability of cancer cells. Recently, ALT has been found to be associated with inactivation of either α-thalassemia/mental retardation syndrome X-linked (ATRX) or death domain-associated (DAXX) protein. In this study, 119 tumors (88 angiosarcomas, 11 epithelioid hemangioendotheliomas, and 20 Kaposi sarcomas) were analyzed to determine the ALT status, its relationship to loss of ATRX/DAXX expression, and the clinicopathological features. In addition, the mutation status in the telomerase reverse transcriptase gene (TERT) promoter was also studied. Loss of ATRX expression was observed in 21% (16/77) of the primary angiosarcomas and 9% (1/11) of epithelioid hemangioendotheliomas. DAXX expression was intact in all but 2 ATRX-deficient angiosarcomas. Telomere-specific fluorescence in situ hybridization assay showed 28% (17/61) of the primary angiosarcomas were ALT positive. Remarkably, ALT was highly associated with loss of ATRX expression: all but 2 ALT-positive angiosarcomas were ATRX deficient. Notably, hepatic angiosarcomas were frequently ATRX deficient (8/13) and/or ALT positive (8/12). None of the secondary angiosarcomas were ATRX/DAXX deficient or ALT positive. The only ATRX-deficient epithelioid hemangioendothelioma was positive for ALT. Forty-seven angiosarcomas were tested for TERT promoter mutation. Despite the fact that angiosarcoma occurs most commonly in sun-damaged skin, mutation was detected in only 1 radiation-associated angiosarcoma (2%). We conclude that ALT is an important telomere maintenance mechanism in primary angiosarcomas. This feature is highly associated with loss of ATRX expression and is frequently observed in hepatic angiosarcomas.


TRBP recruits the Dicer complex to Ago2 for microRNA processing and gene silencing.
MicroRNAs (miRNAs) are generated by a two-step processing pathway to yield RNA molecules of approximately 22 nucleotides that negatively regulate target gene expression at the post-transcriptional level. Primary miRNAs are processed to precursor miRNAs (pre-miRNAs) by the Microprocessor complex. These pre-miRNAs are cleaved by the RNase III Dicer to generate mature miRNAs that direct the RNA-induced silencing complex (RISC) to messenger RNAs with complementary sequence. Here we show that TRBP (the human immunodeficiency virus transactivating response RNA-binding protein), which contains three double-stranded, RNA-binding domains, is an integral component of a Dicer-containing complex. Biochemical analysis of TRBP-containing complexes revealed the association of Dicer-TRBP with Argonaute 2 (Ago2), the catalytic engine of RISC. The physical association of Dicer-TRBP and Ago2 was confirmed after the isolation of the ternary complex using Flag-tagged Ago2 cell lines. In vitro reconstitution assays demonstrated that TRBP is required for the recruitment of Ago2 to the small interfering RNA (siRNA) bound by Dicer. Knockdown of TRBP results in destabilization of Dicer and a consequent loss of miRNA biogenesis. Finally, depletion of the Dicer-TRBP complex via exogenously introduced siRNAs diminished RISC-mediated reporter gene silencing. These results support a role of the Dicer-TRBP complex not only in miRNA processing but also as a platform for RISC assembly.


Assessments of wind-energy potential in selected sites from three geopolitical zones in Nigeria: implications for renewable/sustainable rural electrification.
Electricity generation in rural communities is an acute problem militating against socioeconomic well-being of the populace in these communities in developing countries, including Nigeria. In this paper, assessments of wind-energy potential in selected sites from three major geopolitical zones of Nigeria were investigated. For this, daily wind-speed data from Katsina in northern, Warri in southwestern and Calabar in southeastern Nigeria were analysed using the Gumbel and the Weibull probability distributions for assessing wind-energy potential as a renewable/sustainable solution for the country's rural-electrification problems. Results showed that the wind-speed models identified Katsina with higher wind-speed class than both Warri and Calabar that were otherwise identified as low wind-speed sites. However, econometrics of electricity power simulation at different hub heights of low wind-speed turbine systems showed that the cost of electric-power generation in the three study sites was converging to affordable cost per kWh of electric energy from the wind resource at each site. These power simulations identified cost/kWh of electricity generation at Kaduna as €0.0507, at Warri as €0.0774, and at Calabar as €0.0819. These bare positive implications on renewable/sustainable rural electrification in the study sites even as requisite options for promoting utilization of this viable wind-resource energy in the remote communities in the environs of the study sites were suggested.


Membrane localization of membrane type 5 matrix metalloproteinase by AMPA receptor binding protein and cleavage of cadherins.
Matrix metalloproteinases (MMPs) have been proposed to remodel the extracellular environment of neurons. Here, we report that the metalloproteinase membrane-type 5 MMP (MT5-MMP) binds to AMPA receptor binding protein (ABP) and GRIP (glutamate receptor interaction protein), two related postsynaptic density (PSD) PDZ (postsynaptic density-95/Discs large/zona occludens-1) domain proteins that target AMPA receptors to synapses. The MT5-MMP C terminus binds ABP PDZ5 and the two proteins coimmunoprecipitated and colocalized in heterologous cells and neurons. MT5-MMP localized in filopodia at the tips of growth cones in young [2-5 d in vitro (DIV)] cultured embryonic hippocampal neurons, and at synapses in mature (21 DIV) neurons. Its enrichment in synaptosomes also indicated a synaptic localization in the mature brain. Deletion of the PDZ binding site impaired membrane trafficking of MT5-MMP, whereas exogenous ABP splice forms that are associated either with the plasma membrane or with the cytosol, respectively, colocalized with MT5-MMP in synaptic spines or recruited MT5-MMP to intracellular compartments. We show that endogenous MT5-MMP is found in cultured neurons and brain lysates in a proenzyme form that is activated by furin and degraded by auto-proteolysis. We also identify cadherins as MT5-MMP substrates. These results suggest that ABP directs MT5-MMP proteolytic activity to growth cones and synaptic sites in neurons, where it may regulate axon pathfinding or synapse remodeling through proteolysis of cadherins or other ECM or cell adhesion molecules.


Irbit mediates synergy between ca(2+) and cAMP signaling pathways during epithelial transport in mice.
The cyclic adenosine monophosphate (cAMP) and Ca(2+) signaling pathways synergize to regulate many physiological functions. However, little is known about the mechanisms by which these pathways interact. We investigated the synergy between these signaling pathways in mouse pancreatic and salivary gland ducts. We created mice with disruptions in genes encoding the solute carrier family 26, member 6 (Slc26a6(-/-) mice) and inositol 1,4,5-triphosphate (InsP3) receptor-binding protein released with InsP3 (Irbit(-/-)) mice. We investigated fluid secretion by sealed pancreatic ducts and the function of Slc26a6 and the cystic fibrosis transmembrane conductance regulator (CFTR) in HeLa cells and in ducts isolated from mouse pancreatic and salivary glands. Slc26a6 activity was assayed by measuring intracellular pH, and CFTR activity was assayed by measuring Cl(-) current. Protein interactions were determined by immunoprecipitation analyses. Irbit mediated the synergistic activation of CFTR and Slc26a6 by Ca(2+) and cAMP. In resting cells, Irbit was sequestered by InsP3 receptors (IP3Rs) in the endoplasmic reticulum. Stimulation of Gs-coupled receptors led to phosphorylation of IP3Rs, which increased their affinity for InsP3 and reduced their affinity for Irbit. Subsequent weak stimulation of Gq-coupled receptors, which led to production of low levels of IP3, caused dissociation of Irbit from IP3Rs and allowed translocation of Irbit to CFTR and Slc26a6 in the plasma membrane. These processes stimulated epithelial secretion of electrolytes and fluid. These pathways were not observed in pancreatic and salivary glands from Irbit(-/-) or Slc26a6(-/-) mice, or in salivary gland ducts expressing mutant forms of IP3Rs that could not undergo protein kinase A-mediated phosphorylation. Irbit promotes synergy between the Ca(2+) and cAMP signaling pathways in cultured cells and in pancreatic and salivary ducts from mice. Defects in this pathway could be involved in cystic fibrosis, pancreatitis, or Sjögren syndrome.


Aquaporin 0 enhances gap junction coupling via its cell adhesion function and interaction with connexin 50.
Both connexin 50 (Cx50) and aquaporin 0 (AQP0) have important roles in lens development and homeostasis, and their mutations are associated with human congenital cataracts. We have previously shown that Cx50 directly interacts with AQP0. Here, we demonstrate the importance of the Cx50 intracellular loop (IL) domain in mediating the interaction with AQP0 in the lens in vivo. AQP0 significantly increased (~20-30%) the intercellular coupling and conductance of Cx50 gap junctions. However, this increase was not observed when the IL domain was replaced with those from other lens connexins. The Cx50-AQP0 interaction had no effect on Cx50 hemichannel function. A fusion protein containing three extracellular loop domains of AQP0 efficiently blocked the cell-to-cell adhesion of AQP0 and attenuated the stimulatory effect of AQP0 on Cx50 gap junction conductance. These data suggest that the specific interaction between Cx50 and AQP0 enhances the coupling of Cx50 gap junctions, but not hemichannels, through the cell adhesion function of AQP0. This result establishes a physiological role of AQP0 in the functional regulation of gap junction channels.


Multimodal Hippocampal Subfield Grading For Alzheimer's Disease Classification.
Numerous studies have proposed biomarkers based on magnetic resonance imaging (MRI) to detect and predict the risk of evolution toward Alzheimer's disease (AD). Most of these methods have focused on the hippocampus, which is known to be one of the earliest structures impacted by the disease. To date, patch-based grading approaches provide among the best biomarkers based on the hippocampus. However, this structure is complex and is divided into different subfields, not equally impacted by AD. Former in-vivo imaging studies mainly investigated structural alterations of these subfields using volumetric measurements and microstructural modifications with mean diffusivity measurements. The aim of our work is to improve the current classification performances based on the hippocampus with a new multimodal patch-based framework combining structural and diffusivity MRI. The combination of these two MRI modalities enables the capture of subtle structural and microstructural alterations. Moreover, we propose to study the efficiency of this new framework applied to the hippocampal subfields. To this end, we compare the classification accuracy provided by the different hippocampal subfields using volume, mean diffusivity, and our novel multimodal patch-based grading framework combining structural and diffusion MRI. The experiments conducted in this work show that our new multimodal patch-based method applied to the whole hippocampus provides the most discriminating biomarker for advanced AD detection while our new framework applied into subiculum obtains the best results for AD prediction, improving by two percentage points the accuracy compared to the whole hippocampus.


Targeting of αv integrin identifies a core molecular pathway that regulates fibrosis in several organs.
Myofibroblasts are the major source of extracellular matrix components that accumulate during tissue fibrosis, and hepatic stellate cells (HSCs) are believed to be the major source of myofibroblasts in the liver. To date, robust systems to genetically manipulate these cells have not been developed. We report that Cre under control of the promoter of Pdgfrb (Pdgfrb-Cre) inactivates loxP-flanked genes in mouse HSCs with high efficiency. We used this system to delete the gene encoding α(v) integrin subunit because various α(v)-containing integrins have been suggested as central mediators of fibrosis in multiple organs. Such depletion protected mice from carbon tetrachloride-induced hepatic fibrosis, whereas global loss of β₃, β₅ or β₆ integrins or conditional loss of β₈ integrins in HSCs did not. We also found that Pdgfrb-Cre effectively targeted myofibroblasts in multiple organs, and depletion of the α(v) integrin subunit using this system was protective in other models of organ fibrosis, including pulmonary and renal fibrosis. Pharmacological blockade of α(v)-containing integrins by a small molecule (CWHM 12) attenuated both liver and lung fibrosis, including in a therapeutic manner. These data identify a core pathway that regulates fibrosis and suggest that pharmacological targeting of all α(v) integrins may have clinical utility in the treatment of patients with a broad range of fibrotic diseases.


Separating touching objects in remote sensing imagery: the restricted growing concept and implementations.
This paper defines the restricted growing concept (RGC) fur object separation and provides an algorithmic analysis of its implementations. Our concept decomposes the problem of object separation into two stages. First, separation is achieved by shrinking the objects to their cores while keeping track of their originals as masks. Then the core is grown within the masks obeying the guidelines of a restricted growing algorithm. In this paper, we apply RGC to the remote sensing domain, particularly the synthetic aperture radar (SAR) sea ice images.


Feeding mechanisms in carp: crossflow filtration, palatal protrusions and flow reversals.
It has been hypothesized that, when engulfing food mixed with inorganic particles during benthic feeding, cyprinid fish use protrusions of tissue from the palatal organ to retain the food particles while the inorganic particles are expelled from the opercular slits. In crossflow filtration, the particle suspension is pumped parallel to the filter surface as filtrate exits through the filter pores, causing the suspension to become more concentrated as it travels downstream along the filter. We used high-speed video endoscopy to determine whether carp Cyprinus carpio use crossflow filtration and/or palatal protrusions during benthic feeding. We found that carp use crossflow filtration to concentrate small food particles in the pharyngeal cavity while expelling small dense inorganic particles through the opercular slits and via spits. Our results suggest that, during feeding on small food particles, palatal protrusions serve a localized chemosensory function rather than a mechanical particle-sorting function. However, palatal protrusions did retain large food particles while large inorganic particles were spit anteriorly from the mouth. We also investigated whether flow is continuous and unidirectional during suspension feeding in carp. As reported previously for ventilation in hedgehog skates and for certain industrial crossflow filtration applications, we observed that flow is pulsatile and bidirectional during feeding. These results have implications for hydrodynamic models of crossflow filtration in suspension-feeding fishes.


Feeling Normal? Long-Term Follow-up of Patients with a Cleft Lip-Palate after Rhinoplasty with the Derriford Appearance Scale (DAS-59).
The stigma of nasal deformity due to a congenital cleft lip-palate has an undeniable influence on the affected patient's life. It is therefore of interest to investigate if efforts to reduce esthetic and functional impairments by rhinoplasty (single or multiple) can result in an increased satisfaction with appearance and a self-perception similar to the noncleft population. Retrospective scoring before and after rhinoplasty using the validated Derriford Appearance Scale (DAS-59) and subsequent statistical evaluation and comparison to datasets available in the literature for further classification was used. Of the 61 patients who underwent at least one rhinoplasty, 26 responded to all questions. The mean age of responders was approximately 30 years of age and the male:female ratio was 1:1.2. The scale showed a significant overall improvement after surgery. The full scale and all subscale scores of the DAS-59 were significantly reduced after surgery demonstrating an improvement in the respective categories. Most importantly, if postoperative results were compared with a population concerned and unconcerned about appearance, no difference "facial self-consciousness" of appearance was apparent. Also postoperative subscores for "general self-consciousness" (GSC) and "social self-consciousness" of appearance (SSC) showed no difference from those obtained from the population concerned about appearance. The postoperative subscore for "sexual and bodily self-consciousness" of appearance (SBSC) indicated improvement beyond the level found in the concerned control population. Due to only a low improvement in the difference compared with the subscore representing a "negative self-concept," a statistically significant difference to the concerned population remained, possibly indicating that therapy beyond surgery is needed for improvement. After rhinoplasty, the investigated group of cleft lip-palate patients with nasal deformities showed an improvement in their self-conceived appearance as measured by the DAS-59. Their assessment of self-appearance was comparable to that of a group of noncleft persons with concern about their appearance. Taken together, rhinoplasties, primary and revision, add to the psychosocial well-being and an improved self-perception enhancing quality of life and enabling a more normal life. Further research is needed to clarify how the low reduction found in the "negative self-concept" may be addressed successfully.


Are children as relatives our responsibility? - How nurses perceive their role in caring for children as relatives of seriously ill patients.
The purpose of this study was to elucidate how nurses perceive their role in caring for children as relatives to a parent with a serious physical illness. A qualitative explorative design with focus group interviews was used. In total, 22 nurses working at one neurological, one haematological and two oncological wards were interviewed. The transcripts from the interviews were analysed in steps in accordance with inductive qualitative content analysis. This study revealed six variations in how nurses perceived their role in the encounter with child relatives, ranging from being convinced that it is not their responsibility to being aware of the children's situation and working systematically to support them. Nurses should consider whether their patients have children who might be affected by their parent's illness. The nurses' self-confidence when meeting these children must be increased by education in order to strengthen their professional role. Furthermore, guidelines on how to encounter child relatives are required.


Frequency of infections associated with implanted systems vs cuffed, tunneled Silastic venous catheters in patients with acute leukemia.
A total of 75 central venous catheters were used for prolonged chemotherapy in 39 children with acute lymphocytic leukemia and 21 patients with acute myelocytic leukemia. Infection rates were 2.2 per 100 catheter days with the use of cuffed, tunneled, single-lumen Silastic catheters, 2.0 per 1000 catheter days with cuffed, tunneled, double-lumen Silastic catheters, and 0.5 per 1000 catheter days with the use of implanted venous access systems. Eighty-one percent of catheter sepsis episodes were successfully treated without removal of the catheter. All tunnel infections required withdrawal of the catheter for cure. The microorganisms were gram-positive bacteria in 15%, gram-negative bacteria in 7%, and fungi in 4%. Coagulase-negative staphylococci and Pseudomonas aeruginosa were the most commonly isolated organisms. Three of six fatal sepsis episodes were caused by disseminated fungal infections. We conclude that the use of intracorporeal venous catheter systems in patients with acute lymphocytic leukemia is associated with a lower infection rate than that with cuffed, tunneled Silastic single- or double-lumen catheters and that most septicemias can be cured with antimicrobial therapy without removal of the catheter.


Towards zero training for brain-computer interfacing.
Electroencephalogram (EEG) signals are highly subject-specific and vary considerably even between recording sessions of the same user within the same experimental paradigm. This challenges a stable operation of Brain-Computer Interface (BCI) systems. The classical approach is to train users by neurofeedback to produce fixed stereotypical patterns of brain activity. In the machine learning approach, a widely adapted method for dealing with those variances is to record a so called calibration measurement on the beginning of each session in order to optimize spatial filters and classifiers specifically for each subject and each day. This adaptation of the system to the individual brain signature of each user relieves from the need of extensive user training. In this paper we suggest a new method that overcomes the requirement of these time-consuming calibration recordings for long-term BCI users. The method takes advantage of knowledge collected in previous sessions: By a novel technique, prototypical spatial filters are determined which have better generalization properties compared to single-session filters. In particular, they can be used in follow-up sessions without the need to recalibrate the system. This way the calibration periods can be dramatically shortened or even completely omitted for these 'experienced' BCI users. The feasibility of our novel approach is demonstrated with a series of online BCI experiments. Although performed without any calibration measurement at all, no loss of classification performance was observed.


Evidence for co-evolution of West Nile Virus and house sparrows in North America.
West Nile virus (WNV) has been maintained in North America in enzootic cycles between mosquitoes and birds since it was first described in North America in 1999. House sparrows (HOSPs; Passer domesticus) are a highly competent host for WNV that have contributed to the rapid spread of WNV across the U.S.; however, their competence has been evaluated primarily using an early WNV strain (NY99) that is no longer circulating. Herein, we report that the competence of wild HOSPs for the NY99 strain has decreased significantly over time, suggesting that HOSPs may have developed resistance to this early WNV strain. Moreover, recently isolated WNV strains generate higher peak viremias and mortality in contemporary HOSPs compared to NY99. These data indicate that opposing selective pressures in both the virus and avian host have resulted in a net increase in the level of host competence of North American HOSPs for currently circulating WNV strains.


Risk factors for ductal, lobular, and mixed ductal-lobular breast cancer in a screening population.
Biological distinctions between histologic subtypes of breast cancer suggest etiologic differences, although few studies have been powered to examine such differences. We compared associations between several factors and risk of ductal, lobular, and mixed ductal-lobular breast cancers. We used risk factor data from the Breast Cancer Surveillance Consortium for 3,331,744 mammograms on 1,211,238 women, including 19,119 women diagnosed with invasive breast cancer following mammography (n = 14,818 ductal, 1,602 lobular, and 1,601 mixed ductal-lobular). Histologic subtype-specific risk factor associations were evaluated using Cox regression. Significant positive associations with family history and breast density were similar across subtypes. Hormone therapy use was associated with increased risk of all subtypes, but was most strongly associated with lobular cancer [hazard ratio (HR) = 1.46; 95% confidence interval (CI), 1.25-1.70]. Relative to nulliparous women, parous women had lower risk of ductal and mixed but not lobular cancers (HR = 0.80; 95% CI, 0.76-0.84; HR = 0.79; 95% CI, 0.68-0.93; HR = 0.96; 95% CI, 0.81-1.15, respectively). Late age at first birth was associated with increased risk of all subtypes. Similarities in risk factor associations with ductal, lobular, and mixed breast cancer subtypes were more pronounced than differences. Distinctions between subtype-specific associations were limited to analyses of hormone therapy use and reproductive history. The results of this study indicate that the strongest risk factors for breast cancer overall (that is, family history and breast density) are not histologic subtype specific. Additional studies are needed to better characterize subtype-specific associations with genetic, hormonal, and nonhormonal factors.


The epidemiology of severe Streptococcus pyogenes associated disease in Europe.
Several European countries reported outbreaks of severe disease caused by Streptococcus pyogenes in the late 1980s. This marked a departure from the previous decades, where very few such outbreaks were noted. These changes in disease occurrence formed part of a global phenomenon, the reasons for which have yet to be explained. Results of surveillance activities for invasive S. pyogenes infection within Europe over the past fifteen years identified further increases in many countries. However, variations in surveillance methods between countries preclude robust comparisons being made, illustrating the need for a unified surveillance strategy across Europe. This was finally embodied in the Strep-EURO programme, introduced in 2002.


NTP-CERHR expert panel report on the developmental toxicity of soy infant formula.
Soy infant formula contains soy protein isolates and is fed to infants as a supplement to or replacement for human milk or cow milk. Soy protein isolates contains estrogenic isoflavones (phytoestrogens) that occur naturally in some legumes, especially soybeans. Phytoestrogens are nonsteroidal, estrogenic compounds. In plants, nearly all phytoestrogens are bound to sugar molecules and these phytoestrogen-sugar complexes are not generally considered hormonally active. Phytoestrogens are found in many food products in addition to soy infant formula, especially soy-based foods such as tofu, soy milk, and in some over-the-counter dietary supplements. Soy infant formula was selected for National Toxicology Program (NTP) evaluation because of (1) the availability of large number of developmental toxicity studies in laboratory animals exposed to the isoflavones found in soy infant formula (namely, genistein) or other soy products, as well as few studies on human infants fed soy infant formula, (2) the availability of information on exposures in infants fed soy infant formula, and (3) public concern for effects on infant or child development. On October 2, 2008 (73 FR 57360), the NTP Center for the Evaluation of Risks to Human Reproduction (CERHR) announced its intention to conduct an updated review of soy infant formula to complete a previous evaluation that was initiated in 2005. Both the current and previous evaluations relied on expert panels to assist the NTP in developing its conclusions on the potential developmental effects associated with the use of soy infant formula, presented in the NTP Brief on Soy Infant Formula. The initial expert panel met on March 15 to 17, 2006, to reach conclusions on the potential developmental and reproductive toxicities of soy infant formula and its predominant isoflavone constituent genistein. The expert panel reports were released for public comment on May 5, 2006 (71 FR 28368). On November 8, 2006 (71 FR 65537), CERHR staff released draft NTP Briefs on Genistein and Soy Formula that provided the NTP's interpretation of the potential for genistein and soy infant formula to cause adverse reproductive and/or developmental effects in exposed humans. However, CERHR did not complete these evaluations, finalize the briefs, or issue NTP Monographs on these substances based on this initial evaluation. Between 2006 and 2009, a substantial number of new publications related to human exposure or reproductive and/or developmental toxicity were published for these substances. Thus, CERHR determined that updated evaluations of genistein and soy infant formula were needed. However, the current evaluation focuses only on soy infant formula and the potential developmental toxicity of its major isoflavone components, e.g. genistein, daidzein (and estrogenic metabolite, equol), and glycitein. This updated evaluation does not include an assessment on the potential reproductive toxicity of genistein following exposures during adulthood as was carried out in the 2006 evaluation. CERHR narrowed the scope of the evaluation because the assessment of reproductive effects of genistein following exposure to adults was not considered relevant to the consideration of soy infant formula use in infants during the 2006 evaluation. To obtain updated information about soy infant formula for the CERHR evaluation, the PubMed (Medline) database was searched from February 2006 to August 2009 with genistein/genistin, daidzein/daidzin, glycitein/glycitin, equol, soy, and other relevant keywords. References were also identified from the bibliographies of published literature. The updated expert panel report represents the efforts of a 14-member panel of government and nongovernment scientists, and was prepared with assistance from NTP staff. The finalized report, released on January 15, 2010 (75 FR 2545), reflects consideration of public comments received on a draft report that was released on October 19, 2009, for public comment and discussions that occurred at a public meeting of the expert panel held December 16 to 18, 2009 (74 FR 53509). The finalized report presents conclusions on (1) the strength of scientific evidence that soy infant formula or its isoflavone constituents are developmental toxicants based on data from in vitro, animal, or human studies; (2) the extent of exposures in infants fed soy infant formula; (3) the assessment of the scientific evidence that adverse developmental health effects may be associated with such exposures; and (4) knowledge gaps that will help establish research and testing priorities to reduce uncertainties and increase confidence in future evaluations. The Expert Panel expressed minimal concern for adverse developmental effects in infants fed soy infant formula. This level of concern represents a "2" on the five-level scale of concern used by the NTP that ranges from negligible concern ("1") to serious concern ("5"). The Expert Panel Report on Soy Infant Formula was considered extensively by NTP staff in preparing the 2010 NTP Brief on Soy Infant Formula, which represents the NTP's opinion on the potential for exposure to soy infant formula to cause adverse developmental effects in humans. The NTP concurred with the expert panel that there is minimal concern for adverse effects on development in infants who consume soy infant formula. This conclusion was based on information about soy infant formula provided in the expert panel report, public comments received during the course of the expert panel evaluation, additional scientific information made available since the expert panel meeting, and peer reviewer critiques of the draft NTP Brief by the NTP Board of Scientific Counselors (BSC) on May 10, 2010 (Meeting materials are available at http://ntp.niehs.nih.gov/go/9741.). The BSC voted in favor of the minimal concern conclusion with 7 yes votes, 3 no votes, and 0 abstentions. One member thought that the conclusion should be negligible concern and two members thought that the level of concern should be higher than minimal concern. The NTP's response to the May 10, 2010 review ("peer-review report") is available on the NTP website at http://ntp.niehs.nih.gov/go/9741. The monograph includes the NTP Brief on Soy Infant Formula as well as the entire final Expert Panel Report on Soy Infant Formula. Public comments received as part of the NTP's evaluation of soy infant formula and other background materials are available at http://cerhr.niehs.nih.gov/evals/index.html.


Intestinal threonine utilization for protein and mucin synthesis is decreased in formula-fed preterm pigs.
Threonine is an essential amino acid necessary for synthesis of intestinal (glyco)proteins such as mucin MUC2 to maintain adequate gut barrier function. In premature infants, reduced barrier function may contribute to the development of necrotizing enterocolitis (NEC). Human milk protects against NEC compared with infant formula. Therefore, we hypothesized that formula feeding decreases the MUC2 synthesis rate concomitant with a decrease in intestinal first-pass threonine utilization, predisposing the preterm neonate to NEC. Preterm pigs were delivered by caesarian section and received enteral feeding with formula (FORM; n = 13) or bovine colostrum (COL; n = 6) for 2 d following 48 h of total parenteral nutrition. Pigs received a dual stable isotope tracer infusion of threonine to determine intestinal threonine kinetics. NEC developed in 38% of the FORM pigs, whereas none of the COL pigs were affected (P = 0.13). Intestinal fractional first-pass threonine utilization was lower in FORM pigs (49 ± 2%) than in COL pigs (60 ± 4%) (P = 0.02). In FORM pigs compared with COL pigs, protein synthesis (369 ± 31 mg·kg(-1)·d(-1) vs. 615 ± 54 mg·kg(-1)·d(-1); P = 0.003) and MUC2 synthesis (121 ± 17%/d vs. 184 ± 15%/d; P = 0.02) were lower in the distal small intestine (SI). Our results suggest that formula feeding compared with colostrum feeding in preterm piglets reduces mucosal growth with a concomitant decrease in first-pass splanchnic threonine utilization, protein synthesis, and MUC2 synthesis in the distal SI. Hence, decreased intestinal threonine metabolism and subsequently impaired gut barrier function may predispose the formula-fed infant to developing NEC.


Assessment of Retinal Structural and Functional Characteristics in Eyes with Autoimmune Retinopathy.
To evaluate the thicknesses of individual retinal layers, and the correlation between structural changes and functional loss using spectral domain optical coherence tomography (SD-OCT) scans and electroretinograms (ERG), in eyes with autoimmune retinopathy (AIR). SD-OCT raster scans of 12 eyes from 6 patients serologically diagnosed with AIR were evaluated. Retinal layers were segmented along a 5 mm horizontal scan passing through the fovea. Retinal layers analyzed include full retinal thickness (FRT), retinal pigment epithelium and Bruch's membrane complex (RPE+BM complex), photoreceptor layer (PRL), inner nuclear layer (INL), combined ganglion cell and inner plexiform layers (GCL+), nerve fiber layer (NFL), and combined GCL+ and NFL layers (GCL+/NFL). Changes in the thicknesses of the layers were assessed in 0.5 mm increments along the B-scan in the central, nasal, and temporal regions. These recorded values were compared to corresponding values of 51 eyes from 51 subjects with no known ocular pathology. Full-field ERGs were obtained at corresponding visits and were interpreted by a grader masked to the diagnoses and OCT findings. The mean age of the patients was 59.5 years (range, 33-83), with 4 males (66.6%). Within the control population of 51 subjects, mean age was 51.5 years (range, 40-75), with 25 males (49%). Eyes with AIR showed a loss of retinal tissue compared to eyes with no known ocular pathology at the fovea. Specifically, the FRT, RPE+BM complex, and PRL exhibited thinning of statistically significance. ERG findings demonstrated a functional deficit which showed a good correlation with structural loss. Fifty (50) percent of eyes experienced central photoreceptor (rod and cone) dysfunction and 75% of eyes displayed peripheral photoreceptor (rod and cone) dysfunction. Eyes with AIR show a loss of retinal tissue compared to eyes with no known ocular pathology. The greatest loss appears to occur in the RPE and PRL. ERG findings correlate strongly with the loss of tissue seen in these layers. Thus, therapeutic options may be targeted to preserve these regions of the retina.


Insights into the UDP-sugar selectivities of human UDP-glycosyltransferases (UGT): a molecular modeling perspective.
Enzymes of the human uridine diphosphate (UDP)-glycosyltransferase (UGT) superfamily typically catalyze the covalent addition of a sugar from UDP-sugar cofactors to relatively small lipophilic compounds. The sugar conjugates are often biologically less active with improved water-solubility, facilitating more effective elimination from the body. Experimental data indicate that UGT proteins exhibit differing selectivities toward various UDP-sugars. Although, three-dimensional (3D) structures of UGT proteins are required to provide insights into the UDP-sugar selectivities observed for the various UGT proteins, there are currently, no experimental structures available for human UGTs bound to UDP-sugar(s). Thus, the absence of 3D structures poses a major challenge for analyzing UDP-sugar selectivity at an atomic level. In this commentary, we highlight the application of comparative homology modeling for understanding the UDP-sugar selectivities of UGT proteins. Homology models of the C-terminal (CT) domain indicate a highly conserved structural fold across the UGT family with backbone root mean-squared deviations (rmsds) between 0.066 and 0.079 Å with respect to the UGT2B7-CT X-ray crystal structure. The models show that four residues in the terminal portion of the CT signature sequence play an important role in UDP-sugar selectivity. The N-terminal domain is less likely to be associated with UDP-sugar selectivity, although, a conserved residue, Arg-259 (UGT2B7 numbering) in the UGT 1 and 2 families may influence UDP-sugar selectivity. Overall, the models demonstrate excellent agreement with experimental observations in predicting the key residues that influence the selectivity of UDP-sugar binding.