Alagille syndrome and Wilson disease in siblings: a diagnostic conundrum.
The authors describe two siblings, each with a different, rare genetic condition that affects liver function. The index case, the 18-year-old asymptomatic brother of a young man recently diagnosed with Wilson disease, presented for Wilson disease screening and was also found to have abnormal liver function suggestive of cholestasis. However, ceruloplasmin level, 24 h urine copper concentration and liver synthetic function were normal. Further hepatic investigations and genetic mutation analysis were performed, ultimately leading to a diagnosis of Alagille syndrome. He was treated with ursodiol, which resulted in normalization of his liver function tests. Subsequently, he was found to be a carrier for a mutation in the Wilson disease gene, ATP7B. In the present report, the potential implications of being a heterozygote for Wilson disease in the context of Alagille syndrome are discussed. Also stressed is that care must be exercised by the clinician when diagnosing family members who may present with two different disorders closely mimicking one another.


DNAAF1 links heart laterality with the AAA+ ATPase RUVBL1 and ciliary intraflagellar transport.
DNAAF1 (LRRC50) is a cytoplasmic protein required for dynein heavy chain assembly and cilia motility, and DNAAF1 mutations cause primary ciliary dyskinesia (PCD; MIM 613193). We describe four families with DNAAF1 mutations and complex congenital heart disease (CHD). In three families, all affected individuals have typical PCD phenotypes. However, an additional family demonstrates isolated CHD (heterotaxy) in two affected siblings, but no clinical evidence of PCD. We identified a homozygous DNAAF1 missense mutation, p.Leu191Phe, as causative for heterotaxy in this family. Genetic complementation in dnaaf1-null zebrafish embryos demonstrated the rescue of normal heart looping with wild-type human DNAAF1, but not the p.Leu191Phe variant, supporting the conserved pathogenicity of this DNAAF1 missense mutation. This observation points to a phenotypic continuum between CHD and PCD, providing new insights into the pathogenesis of isolated CHD. In further investigations of the function of DNAAF1 in dynein arm assembly, we identified interactions with members of a putative dynein arm assembly complex. These include the ciliary intraflagellar transport protein IFT88 and the AAA+ (ATPases Associated with various cellular Activities) family proteins RUVBL1 (Pontin) and RUVBL2 (Reptin). Co-localization studies support these findings, with the loss of RUVBL1 perturbing the co-localization of DNAAF1 with IFT88. We show that RUVBL1 orthologues have an asymmetric left-sided distribution at both the mouse embryonic node and the Kupffer's vesicle in zebrafish embryos, with the latter asymmetry dependent on DNAAF1. These results suggest that DNAAF1-RUVBL1 biochemical and genetic interactions have a novel functional role in symmetry breaking and cardiac development.


Continuation tapping to triggered melodies: motor resonance effects of melodic motion.
Common Coding theory predicts that perceived action should resonate in produced action to which it bears some resemblance. Here we show that the qualities of motion commonly attributed to melodies are instantiated in motor plans that control timed movements. Participants attempted to tap a steady beat. Each tap triggered a sounded tone, and successive tones were systematically varied in pitch to form short melodies. Tapping behavior was monitored with motion capture. Although instructed to ignore them, triggered tones systematically affected timing and finger movement. When slower melodic motion was implied by a contour change or a smaller pitch displacement, the interval-tap interval (ITI) was longer. When faster melodic motion was implied by a preserved pitch contour or a larger pitch displacement, ITI was shorter. Kinematic recordings suggested that ITI Error arose from an initial failure to disambiguate perception (i.e., velocity implied by melodic motion) from action (i.e., finger velocity [FV]). Early in the tap trajectory, slower FV was associated with longer ITI and faster FV was associated with shorter ITI. These associations were reversed near mid-trajectory, suggesting a transition from execution of motor planning to online control (Glover et al. in Exp Brain Res 154:103-108, 2004).


Electrocardiogram blood pressure and systolic time intervals long term monitoring in ambulant patients.
Ambulatory monitoring of the intra arterial blood pressure (IBP), through the Selyg-Oxford System, has been used so far primarily for studies on hypertension. Aim of our study is to explore the possibility of obtaining from IBP indications useful to extend its field of application. The study will investigate: the usefulness of the continuous monitoring of the systolic time intervals (STI), measured from IBP as ventricular performance indicators, the increasing of specificity of the ECG allowed by the simultaneous beat by beat estimate of the STI's, the possibility of achieving the same information from non invasive peripheral pressure transducers. In this paper we are presenting the results obtained from the combined analysis of the STI's, ECG and IBP in a first series of 13 patients, selected out of 50 on the basis of the highest number of episodes of IBP variations, to allow the evaluation of the algorithms performances in severe conditions.


Signal processing using artificial neural network for BOTDA sensor system.
We experimentally demonstrate the use of artificial neural network (ANN) to process sensing signals obtained from Brillouin optical time domain analyzer (BOTDA). The distributed temperature information is extracted directly from the local Brillouin gain spectra (BGSs) along the fiber under test without the process of determination of Brillouin frequency shift (BFS) and hence conversion from BFS to temperature. Unlike our previous work for short sensing distance where ANN is trained by measured BGSs, here we employ ideal BGSs with different linewidths to train the ANN in order to take the linewidth variation due to different conditions from the training and testing phases into account, making it feasible for long distance sensing. Moreover, the performance of ANN is compared with other two techniques, Lorentzian curve fitting and cross-correlation method, and our results show that ANN has higher accuracy and larger tolerance to measurement error, especially at large frequency scanning step. We also show that the temperature extraction from BOTDA measurements employing ANN is significantly faster than the other two approaches. Hence ANN can be an excellent alternative tool to process BGSs measured by BOTDA and obtain temperature distribution along the fiber, especially when large frequency scanning step is adopted to significantly reduce the measurement time but without sacrifice of sensing accuracy.


Amyloid-β 11C-PiB-PET imaging results from 2 randomized bapineuzumab phase 3 AD trials.
To evaluate the effects of bapineuzumab on brain β-amyloid (Aβ) burden using (11)C-Pittsburgh compound B ((11)C-PiB)-PET. Two phase 3 clinical trials, 1 each in apolipoprotein APOE ε4 carriers and noncarriers, were conducted in patients with mild to moderate Alzheimer disease dementia. Bapineuzumab, an anti-Aβ monoclonal antibody, or placebo, was administered by IV infusion every 13 weeks for 78 weeks. PET substudies assessed change in brain fibrillar Aβ over 71 weeks using an (11)C-PiB-PET standardized uptake value ratio (SUVr) global cortical average (GCA) comprising the average SUVr from 5 cortical regions of interest with cerebellar gray matter as the reference region. A total of 115 carriers and 39 noncarriers were analyzed. The difference (δ) in mean baseline to 71 week change in (11)C-PiB-PET GCA between bapineuzumab and placebo was significant in carriers (0.5 mg/kg vs placebo δ = -0.101; p = 0.004) and in pooled analyses of both carriers and noncarriers (0.5 mg/kg vs placebo δ = -0.068; p = 0.027; 1.0 mg/kg vs placebo δ = -0.133; p = 0.028) but not in the noncarrier trial separately. Analyses by individual region of interest and in mild disease yielded findings similar to the main trial results. The (11)C-PiB-PET imaging results demonstrated reduction of fibrillar Aβ accumulation in patients with Alzheimer disease treated with bapineuzumab; however, as no clinical benefit was observed, the findings are consistent with the hypotheses that bapineuzumab may not have been initiated early enough in the disease course, the doses were insufficient, or the most critical Aβ species were inadequately targeted.


Incidence of childhood obesity in the United States.
Although the increased prevalence of childhood obesity in the United States has been documented, little is known about its incidence. We report here on the national incidence of obesity among elementary-school children. We evaluated data from the Early Childhood Longitudinal Study, Kindergarten Class of 1998-1999, a representative prospective cohort of 7738 participants who were in kindergarten in 1998 in the United States. Weight and height were measured seven times between 1998 and 2007. Of the 7738 participants, 6807 were not obese at baseline; these participants were followed for 50,396 person-years. We used standard thresholds from the Centers for Disease Control and Prevention to define "overweight" and "obese" categories. We estimated the annual incidence of obesity, the cumulative incidence over 9 years, and the incidence density (cases per person-years) overall and according to sex, socioeconomic status, race or ethnic group, birth weight, and kindergarten weight. When the children entered kindergarten (mean age, 5.6 years), 12.4% were obese and another 14.9% were overweight; in eighth grade (mean age, 14.1 years), 20.8% were obese and 17.0% were overweight. The annual incidence of obesity decreased from 5.4% during kindergarten to 1.7% between fifth and eighth grade. Overweight 5-year-olds were four times as likely as normal-weight children to become obese (9-year cumulative incidence, 31.8% vs. 7.9%), with rates of 91.5 versus 17.2 per 1000 person-years. Among children who became obese between the ages of 5 and 14 years, nearly half had been overweight and 75% had been above the 70th percentile for body-mass index at baseline. Incident obesity between the ages of 5 and 14 years was more likely to have occurred at younger ages, primarily among children who had entered kindergarten overweight. (Funded by the Eunice Kennedy Shriver National Institute of Child Health and Human Development.).


Mixed protein-polysaccharide interfacial layers: a self consistent field calculation study.
Mixed interfacial films of protein and polysaccharide have been investigated using self consistent field (SCF) calculations. The colloidal interactions mediated by such composite layers between two approaching surfaces have been considered. Two types of systems have been studied: (a) covalently-bonded polysaccharide and protein, and (b) the excess presence of polysaccharide at the interface, occurring through electrostatic interaction with an already existing, oppositely charged, protein layer. Our calculations show that for covalently-bonded complexes, depending on the location of the protein-polysaccharide bond, the attachment of short uncharged chains to the protein can be detrimental to provision of repulsive colloidal forces by such complexes. We have attributed this to an increased tendency of the hybrid polymer to adopt bridging configurations in the gap between two nearby surfaces. For larger grafted chains this bridging effect is eliminated, and the expected enhanced steric stabilization of the protein-polysaccharide conjugate is achieved. For adsorbed films formed through electrostatic interactions between these two biopolymers, stronger repulsive forces between the surfaces are produced, at an intermediate level of charging for the polysaccharides. This has been related to a maximum level of adsorption of polysaccharide, as the number of charged segments of the chain is varied. The peak occurs at higher levels of charging as the salt concentration in the bulk solution is increased. We have also observed the experimentally-reported phenomenon of charge overcompensation, arising from adsorption of the polysaccharide chains onto the primary protein layer. The importance of the non-uniform charge distribution of the polysaccharide molecule, in providing an explanation for this effect, has been demonstrated.


Carbachol-induced decrease in thyroid cell adenylyl cyclase activity is independent of calcium and phosphodiesterase activation.
The mechanism of adenylyl cyclase desensitization by carbachol, an agent that stimulates polyphosphoinositide hydrolysis, was studied in thyroid cells. Incubation of cultured dog thyroid cells with 10 microM carbachol for 2-4 hr reduced the subsequent thyrotropic hormone (TSH) stimulation of adenylyl cyclase activity of membrane preparations by approximately 40%. This inhibition was reversed by atropine, occurred even in a Ca(2+)-free medium containing ethylene glycol bis(beta-aminoethylether)-N,N,N',N'-tetraacetic acid, and was not reproduced by the Ca2+ ionophore A23187. The carbachol effect was not prevented by simultaneous incubation of cells with either isobutylmethylxanthine, an inhibitor of phosphodiesterase, or H-7, an inhibitor of protein kinase. Pretreatment of cells with pertussis toxin to inactivate the Gi inhibitory protein also failed to affect the carbachol inhibition. Although carbachol did not reduce the basal or the TSH-stimulated cyclase activities when added to membranes directly during the assay, exposure of cells to carbachol for 2-4 hr resulted in long lasting inhibition of TSH-stimulated cyclase activity (for at least 24 hr); recovery was seen by 48 hr after its removal. Carbachol pretreatment had no effect on 125I-TSH binding to membranes but reduced the cyclase stimulation by not only TSH but also cholera toxin, guanosine 5'-O-(3-thio)triphosphate, and forskolin; it also significantly reduced the cholera toxin-mediated AD[32P]-ribosylation of Gs in membranes. These data indicate that carbachol-induced inhibition of adenylyl cyclase occurs beyond the level of TSH receptor binding and that Gs is a possible site of its action. Thus, in dog thyroid cells, carbachol, via muscarinic receptors, can reduce the adenylyl cyclase activity by a process that does not involve Ca2+ or activation of phosphodiesterase.


Inherent homogenous media dispersion compensation in frequency domain optical coherence tomography by accurate k-sampling.
Depth dependent broadening of the axial point spread function due to dispersion in the imaged media, and algorithms for postprocess correction, have been previously described for both time domain and frequency domain optical coherence tomography. We show that homogeneous media dispersion artifacts disappear when frequency domain samples are acquired with uniform spacing in circular wavenumber, as opposed to uniform sampling in optical frequency. We further explicate the source of this point spread broadening and simulate its magnitude in aqueous media. We experimentally demonstrate media dispersion compensation in high dispersion glass by choosing sample frequencies at equal intervals of media index of refraction divided by vacuum wavelength, and we recover unbroadened reflections without an additional postprocessing step.


Metabolic disturbances associated with systemic lupus erythematosus.
The metabolic disturbances that underlie systemic lupus erythematosus are currently unknown. A metabolomic study was executed, comparing the sera of 20 SLE patients against that of healthy controls, using LC/MS and GC/MS platforms. Validation of key differences was performed using an independent cohort of 38 SLE patients and orthogonal assays. SLE sera showed evidence of profoundly dampened glycolysis, Krebs cycle, fatty acid β oxidation and amino acid metabolism, alluding to reduced energy biogenesis from all sources. Whereas long-chain fatty acids, including the n3 and n6 essential fatty acids, were significantly reduced, medium chain fatty acids and serum free fatty acids were elevated. The SLE metabolome exhibited profound lipid peroxidation, reflective of oxidative damage. Deficiencies were noted in the cellular anti-oxidant, glutathione, and all methyl group donors, including cysteine, methionine, and choline, as well as phosphocholines. The best discriminators of SLE included elevated lipid peroxidation products, MDA, gamma-glutamyl peptides, GGT, leukotriene B4 and 5-HETE. Importantly, similar elevations were not observed in another chronic inflammatory autoimmune disease, rheumatoid arthritis. To sum, comprehensive profiling of the SLE metabolome reveals evidence of heightened oxidative stress, inflammation, reduced energy generation, altered lipid profiles and a pro-thrombotic state. Resetting the SLE metabolome, either by targeting selected molecules or by supplementing the diet with essential fatty acids, vitamins and methyl group donors offers novel opportunities for disease modulation in this disabling systemic autoimmune ailment.


Solutions to the cost-of-selection dilemma.
There are various biologically realistic ways around the cost-of-selection restriction on rapid multiple-gene substitution. Some of these ways depend upon particular forms of interaction or linkage in the genes undergoing substitution; other ways depend on particular conditions of population size and structure. The special genotypic and populational conditions required for rapid evolutionary change in genetically complex characters are not unusual in higher organisms.


Carboxy Mb at pH 3. Time-resolved resonance Raman study at cryogenic temperatures.
Cryogenic samples of MbCO at pH3 are studied using nanosecond and picosecond time-resolved resonance Raman spectroscopy. It is observed that under excitation conditions sufficient to completely photodissociate MbCO at pH7, the pH3 sample at 10 ns remains substantially unphotolyzed even at 15 K. The similarity in the optical and resonance Raman spectra of MbCO at pH3 with that of pH7 indicates that at pH3 the iron remains six-coordinate and low-spin. The Fe-CO stretch frequency is consistent with a more upright CO orientation. The absence of the v(Fe-His) band in the 30 ps photoproduct Raman spectrum suggests that the Fe-His(F8) bond is broken within 30 ps of photodissociation. Other Raman bands, though, are not consistent with a normal four-coordinate heme for the photoproduct, Mb*. Suggested possible interpretations include a four-coordinate heme highly perturbed by the close lying protonated proximal histidine or a five-coordinate heme with the Fe-His bond significantly weakened. The partial photolysis monitored at 30 ps and 100 K indicates either a significant amount of geminate recombination within 30 ps or low quantum yield or photolysis. The time course for CO recombination is monitored via the Raman spectra from 30 ps to 3 ns at 100 K and 160 K. Of the fraction of protein-ligand pairs that remain photodissociated at 30 ps, 50% recombine by approximately 250 ps at 100 K and 160 K, supporting the flash photolysis rebinding data of Cowen et al. (Cowen, B. R. 1990. Ph. D. thesis. University of Illinois at Urbana-Champaign; Cowen, B. R., D. Braunstein, H. Frauenfelder, P. J. Steinbach, and R. D. Young. 1989. Biophys. J. 55:55a. [Abstr.].) The conclusions from these resonance Raman studies are extended to solution phase studies at ambient temperatures.


Urinary oncofetal ED-A fibronectin correlates with poor prognosis in patients with bladder cancer.
The extracellular matrix protein fibronectin (FN) contributes to the structural integrity of tissues as well as the adhesive and migratory functions of cells. While FN is abundantly expressed in adult tissues, the expression of several alternatively spliced FN isoforms is restricted to embryonic development, tissue remodeling and cancer. These FN isoforms, designated ED-A and ED-B, are frequently expressed by cancer cells, tumor-associated fibroblasts and newly forming blood vessels. Using a highly sensitive collagen-based indirect ELISA, we evaluated the correlation of urinary ED-A and ED-B at time of cystectomy with overall survival in patients with high-grade bladder cancer (BCa). Detectable levels of total FN as well as ED-A and ED-B were found in urine from 85, 73 and 51 % of BCa patients, respectively. The presence of urinary ED-A was a significant independent predictor of 2-year overall survival (OS) after adjusting for age, tumor stage, lymph node stage, and urinary creatinine by multivariable Logistic Regression (p = 0.029, OR = 4.26, 95 % CI 1.16-15.71) and improved accuracy by 3.6 %. Furthermore, detection of ED-A in the urine was a significant discriminator of survival specifically in BCa patients with negative lymph node status (Log-Rank, p = 0.006; HR = 5.78, 95 % CI 1.39-24.13). Lastly, multivariable Cox proportional hazards analysis revealed that urinary ED-A was an independent prognostic indicator of 5-year OS rate for patients with BCa (p = 0.04, HR = 2.20, 95 % CI 1.04-4.69). Together, these data suggest that cancer-derived, alternatively spliced FN isoforms can act as prognostic indicators and that additional studies are warranted to assess the clinical utility of ED-A in BCa.


A data-driven study of Chinese participants' social judgments of Chinese faces.
Social judgments of faces made by Western participants are thought to be underpinned by two dimensions: valence and dominance. Because some research suggests that Western and Eastern participants process faces differently, the two-dimensional model of face evaluation may not necessarily apply to judgments of faces by Eastern participants. Here we used a data-driven approach to investigate the components underlying social judgments of Chinese faces by Chinese participants. Analyses showed that social judgments of Chinese faces by Chinese participants are partly underpinned by a general approachability dimension similar to the valence dimension previously found to underpin Western participants' evaluations of White faces. However, we found that a general capability dimension, rather than a dominance dimension, contributed to Chinese participants' evaluations of Chinese faces. Thus, our findings present evidence for both cultural similarities and cultural differences in social evaluations of faces. Importantly, the dimension that explained most of the variance in Chinese participants' social judgments of faces was strikingly similar to the valence dimension previously reported for Western participants.


Intrinsic conflicts of interest in clinical research: a need for disclosure.
Protection of human subjects from investigators' conflicts of interest is critical to the integrity of clinical investigation. Personal financial conflicts of interest are addressed by university policies, professional society guidelines, public standards, and government regulation, but "intrinsic conflict of interest"--conflicts of interest inherent in all clinical research--have received relatively less attention. Such conflicts arise in all clinical research endeavors as a result of the tension among professionals' responsibilities to their research and to their patients and both academic and financial incentives. These conflicts should be disclosed to research subjects and managed as assiduously as are financial conflicts of interest.


Defective synaptic transmission causes disease signs in a mouse model of juvenile neuronal ceroid lipofuscinosis.
Juvenile neuronal ceroid lipofuscinosis (JNCL or Batten disease) caused by mutations in the CLN3 gene is the most prevalent inherited neurodegenerative disease in childhood resulting in widespread central nervous system dysfunction and premature death. The consequences of CLN3 mutation on the progression of the disease, on neuronal transmission, and on central nervous network dysfunction are poorly understood. We used Cln3 knockout (Cln3Δex1-6) mice and found increased anxiety-related behavior and impaired aversive learning as well as markedly affected motor function including disordered coordination. Patch-clamp and loose-patch recordings revealed severely affected inhibitory and excitatory synaptic transmission in the amygdala, hippocampus, and cerebellar networks. Changes in presynaptic release properties may result from dysfunction of CLN3 protein. Furthermore, loss of calbindin, neuropeptide Y, parvalbumin, and GAD65-positive interneurons in central networks collectively support the hypothesis that degeneration of GABAergic interneurons may be the cause of supraspinal GABAergic disinhibition.


Prevalence of FOXC1 Variants in Individuals With a Suspected Diagnosis of Primary Congenital Glaucoma.
Both primary and secondary forms of childhood glaucoma have many distinct causative mechanisms, and in many cases a cause is not immediately clear. The broad phenotypic spectrum of secondary glaucoma, particularly in individuals with variants in FOXC1 or PITX2 genes associated with Axenfeld-Rieger syndrome, makes it more difficult to diagnose patients with milder phenotypes. These cases are occasionally classified and managed as primary congenital glaucoma. To investigate the prevalence of FOXC1 variants in participants with a suspected diagnosis of primary congenital glaucoma. Australian and Italian cohorts were recruited from January 1, 2007, through March 1, 2016. Australian individuals were recruited through the Australian and New Zealand Registry of Advanced Glaucoma and Italian individuals through the Genetic and Ophthalmology Unit of l'Azienda Socio-Sanitaria Territoriale Grande Ospedale Metropolitano Niguarda in Milan, Italy. We performed exome sequencing, in combination with Sanger sequencing and multiplex ligation-dependent probe amplification, to detect variants of FOXC1 in individuals with a suspected diagnosis of primary congenital glaucoma established by their treating specialist. Data analysis was completed from June 2015 to November 2017. Identification of single-nucleotide and copy number variants in FOXC1, along with phenotypic characterization of the individuals who carried them. A total of 131 individuals with a suspected diagnosis of primary congenital glaucoma were included. The mean (SD) age at recruitment in the Australian cohort was 24.3 (18.1) years; 37 of 84 Australian participants (44.0%) were female, and 71 of 84 (84.5%) were of European ancestry. The mean (SD) age at recruitment was 22.5 (18.4) years in the Italian cohort; 21 of 47 Italian participants (44.7%) were female, and 45 of 47 (95.7%) were of European ancestry. We observed rare, predicted deleterious FOXC1 variants in 8 of 131 participants (6.1%), or 8 of 166 participants (4.8%) when including those explained by variants in CYP1B1. On reexamination or reinvestigation, all of these individuals had at least 1 detectable ocular and/or systemic feature associated with Axenfeld-Rieger syndrome. These data highlight the genetic and phenotypic heterogeneity of childhood glaucoma and support the use of gene panels incorporating FOXC1 as a diagnostic aid, especially because clinical features of Axenfeld-Rieger syndrome can be subtle. Further replication of these results will be needed to support the future use of such panels.


Comparative functional genomics of the fission yeasts.
The fission yeast clade--comprising Schizosaccharomyces pombe, S. octosporus, S. cryophilus, and S. japonicus--occupies the basal branch of Ascomycete fungi and is an important model of eukaryote biology. A comparative annotation of these genomes identified a near extinction of transposons and the associated innovation of transposon-free centromeres. Expression analysis established that meiotic genes are subject to antisense transcription during vegetative growth, which suggests a mechanism for their tight regulation. In addition, trans-acting regulators control new genes within the context of expanded functional modules for meiosis and stress response. Differences in gene content and regulation also explain why, unlike the budding yeast of Saccharomycotina, fission yeasts cannot use ethanol as a primary carbon source. These analyses elucidate the genome structure and gene regulation of fission yeast and provide tools for investigation across the Schizosaccharomyces clade.


Surfaceome profiling enables isolation of cancer-specific exosomal cargo in liquid biopsies from pancreatic cancer patients.
Detection of circulating tumor DNA can be limited due to their relative scarcity in circulation, particularly while patients are actively undergoing therapy. Exosomes provide a vehicle through which cancer-specific material can be enriched from the compendium of circulating non-neoplastic tissue-derived nucleic acids. We carried out a comprehensive profiling of the pancreatic ductal adenocarcinoma (PDAC) exosomal 'surfaceome' in order to identify surface proteins that will render liquid biopsies amenable to cancer-derived exosome enrichment for downstream molecular profiling. Surface exosomal proteins were profiled in 13 human PDAC and 2 non-neoplastic cell lines by liquid chromatography-mass spectrometry. A total of 173 prospectively collected blood samples from 103 PDAC patients underwent exosome isolation. Droplet digital PCR was used on 74 patients (136 total exosome samples) to determine baseline KRAS mutation call rates while patients were on therapy. PDAC-specific exosome capture was then carried out on additional 29 patients (37 samples) using an antibody cocktail directed against selected proteins, followed by droplet digital PCR analysis. Exosomal DNA in a PDAC patient resistant to therapy were profiled using a molecular barcoded, targeted sequencing panel to determine the utility of enriched nucleic acid material for comprehensive molecular analysis. Proteomic analysis of the exosome 'surfaceome' revealed multiple PDAC-specific biomarker candidates: CLDN4, EPCAM, CD151, LGALS3BP, HIST2H2BE, and HIST2H2BF. KRAS mutations in total exosomes were detected in 44.1% of patients undergoing active therapy compared with 73.0% following exosome capture using the selected biomarkers. Enrichment of exosomal cargo was amenable to molecular profiling, elucidating a putative mechanism of resistance to PARP inhibitor therapy in a patient harboring a BRCA2 mutation. Exosomes provide unique opportunities in the context of liquid biopsies for enrichment of tumor-specific material in circulation. We present a comprehensive surfaceome characterization of PDAC exosomes which allows for capture and molecular profiling of tumor-derived DNA.


Global issues and opportunities for optimized retinoblastoma care.
The RB1 gene is important in all human cancers. Studies of human retinoblastoma point to a rare retinal cell with extreme dependency on RB1 for initiation but not progression to full malignancy. In developed countries, genetic testing within affected families can predict children at high risk of retinoblastoma before birth; chemotherapy with local therapy often saves eyes and vision; and mortality is 4%. In less developed countries where 92% of children with retinoblastoma are born, mortality reaches 90%. Global collaboration is building for the dramatic change in mortality that awareness, simple expertise and therapies could achieve in less developed countries.


An integrated framework for the geographic surveillance of chronic disease.
Geographic public health surveillance is concerned with describing and disseminating geographic information about disease and other measures of health to policy makers and the public. While methodological developments in the geographical analysis of disease are numerous, few have been integrated into a framework that also considers the effects of case ascertainment bias on the effectiveness of chronic disease surveillance. We present a framework for the geographic surveillance of chronic disease that integrates methodological developments in the spatial statistical analysis and case ascertainment. The framework uses an hierarchical approach to organize and model health information derived from an administrative health data system, and importantly, supports the detection and analysis of case ascertainment bias in geographic data. We test the framework on asthmatic data from Alberta, Canada. We observe high prevalence in south-western Alberta, particularly among Aboriginal females. We also observe that persons likely mistaken for asthmatics tend to be distributed in a pattern similar to asthmatics, suggesting that there may be an underlying social vulnerability to a variety of respiratory illnesses, or the presence of a diagnostic practice style effect. Finally, we note that clustering of asthmatics tends to occur at small geographic scales, while clustering of persons mistaken for asthmatics tends to occur at larger geographic scales. Routine and ongoing geographic surveillance of chronic diseases is critical to developing an understanding of underlying epidemiology, and is critical to informing policy makers and the public about the health of the population.


Information for genetic management of mtDNA disease: sampling pathogenic mtDNA mutants in the human germline and in placenta.
Families with a child who died of severe, maternally inherited mitochondrial DNA (mtDNA) disease need information on recurrence risk. Estimating this risk is difficult because of (a) heteroplasmy-the coexistence of mutant and normal mtDNA in the same person-and (b) the so-called mitochondrial bottleneck, whereby the small number of mtDNAs that become the founders for the offspring cause variation in dose of mutant mtDNA. The timing of the bottleneck and of segregation of mtDNA during foetal life determines the management options. Therefore, mtDNA heteroplasmy was studied in oocytes and placenta of women in affected families. One mother of a child dying from Leigh syndrome due to the 9176T-->C mtDNA mutation transmitted various loads of mutant mtDNA to < or =3 of 20 oocytes. This was used to estimate recurrence as < or =5%. She subsequently conceived a healthy son naturally. Analysis of the placenta showed that some segregation also occurred during placental development, with the mutant mtDNA load varying by >10% in a placenta carrying 65% 3243A-->G mutant mtDNA. This is the first report of (a) an oocyte analysis for preconception counselling, specifically, refining recurrence risks of rare mutations and (b) a widely different load of a pathogenic mtDNA mutation in multiple oocytes, apparently confined to the germline, in an asymptomatic carrier of an mtDNA disease. This suggests that a major component of the bottleneck occurs during oogenesis, probably early in the foetal life of the mother. The variable mutant load in placenta implies that estimates based on a single sample in prenatal diagnosis of mtDNA disorders have limited accuracy.


From granuloma to fibrosis in interstitial lung diseases: molecular and cellular interactions.
Granuloma is a feature of many chronic interstitial lung diseases, and may serve as a focus for subsequent fibrosis. Granulomas are composed of structured masses of cells of the macrophage lineage, which adopt an epithelioid aspect, interspersed with lymphocytes. They are formed around local centres of irritation. During their resolution, fibroblasts congregate around the structures and may penetrate the interior. In many cases, granulomas can disappear without leaving lasting traces. However, especially when damage has occurred to the surrounding tissue, permanent scarring and fibrosis may occur. Both types of cell present in the granuloma are capable of secreting a number of factors influencing the accumulation and proliferation of fibroblasts, both positively and negatively. The possible roles played by the different factors and, especially, interactions between them are discussed in the light of fibrosis formation. Possible therapeutic interventions are summarized.


Novel non-occlusive temporary endoluminal neck protection device to assist in the treatment of aneurysms in a canine model.
Current models for the endovascular treatment of aneurysms often require the use of adjuvant devices. Balloon-assisted techniques require temporary occlusion of the parent vessel. Intravascular stents require the use of antiplatelet medication. A novel device that achieves temporary neck protection without parent vessel occlusion or antiplatelet medication is examined in a canine model. 20 sidewall wide-necked aneurysms were created in 10 canines, one in each carotid artery, using a vein graft technique, and allowed to mature. In each canine, one aneurysm was catheterized with a microcatheter while the neck protection device (NPD) was unsheathed across the neck of the aneurysm and the aneurysm was coiled. The second aneurysm in each canine was coiled without an adjunctive device. The NPD was assessed for coil herniation, coil entrapment within the device, deployability and retrievability. Five animals were killed immediately and five were killed at 28 days and the carotid artery was explanted and sent for necropsy to assess for injury to the endothelium. All aneurysms were successfully treated with no adverse events. The NPD was well visualized and unsheathed and resheathed successfully in all cases. There was no coil entrapment within the device, and there were no cases of coil herniation. There was no evidence of endothelial injury or dissection. The device was technically successful in all devices and further development of this device may be of benefit in endovascular treatment of aneurysms.


Transmission of Anaplasma phagocytophila (human granulocytic ehrlichiosis agent) in horses using experimentally infected ticks (Ixodes scapularis).
Most human granulocytic ehrlichiosis (HGE) studies carried out in horses use needle inoculation of infected leucocytes or cell cultures. This route of inoculation does not accurately reflect natural infection of the tick-borne agent. To investigate whether tick transmission influences the course of granulocytic ehrlichiosis in the horse model, experimental transmission through infected laboratory-reared Ixodes scapularis ticks was attempted into two healthy horses. One additional horse served as negative control and was exposed to uninfected ticks. Eleven days after exposure to nymphal or adult ticks infected with Anaplasma phagocytophila (HGE agent) the two horses developed severe clinical and laboratory signs consistent with granulocytic ehrlichiosis. Bacteraemia was determined at various time points in the two horses by observation of morulae within neutrophils and by detection of A. phagocytophila genomic DNA by PCR of peripheral blood leucocytes. Further, both horses seroconverted. In contrast the control horse stayed uninfected. The results demonstrate that A. phagocytophila can be experimentally transmitted by infected nymphal and adult ticks and that the agent is able to produce a severe disease, similar to naturally occurring cases. Therefore, tick transmission is highly reproducible and can be successfully used in the equine animal model in order to study HGE.


Gait abnormalities following resection of talocalcaneal coalition.
Eleven patients (fourteen feet) had resection of a coalition of the middle facet of the talocalcaneal joint because of symptoms that had been present for a mean of four years (range, 0.5 to nine years). The mean age at the time of the resection was seventeen years (range, thirteen to thirty-two years). Nine patients were male and two were female. Five feet had resection of the bone bridge with interposition of fat or tendon, and nine had resection without any interposed material. The mean duration of follow-up was six years (range, two to thirteen years). The clinical result was excellent for five feet, good for four, fair for three, and poor for two. Inversion was 8 +/- 4.5 degrees (mean and standard deviation) on the involved side and 17 +/- 6.2 degrees on the uninvolved side (p = 0.002). Three feet later had evidence of osteoarthrosis of the subtalar joint. Gait analysis demonstrated decreased motion of the hindfoot and the ankle, compared with that in normal subjects and with that in the contralateral (uninvolved) foot, in the sagittal and coronal planes during walking on a level surface. Motion was also reduced, particularly in the sagittal and coronal planes, during walking on a side slope. Ground-reaction forces (F8 and F9), temporal force factors (T2, T3, and T7), and the percentage of the gait cycle that consisted of the stance phase on the side-sloping walkway were different from those in normal subjects or in the uninvolved foot. These data indicate that, although most of our patients who had resection of a talocalcaneal coalition had a successful clinical result, most had a residual functional deficit.


Family carers in stroke care: examining the relationship between problem-solving, depression and general health.
The purpose of this article is to describe the problem-solving abilities of Hong Kong family carers looking after a stroke patients at home and report the relationships between their perceived problem-solving abilities with their depression level, general health status, and the functional recovery of stroke patients. Previous research on supportive interventions for caregiving in stroke care suggests that enhancing carers' problem-solving abilities is useful. Nevertheless, not much is known about the relationship between carers' problem-solving abilities and their physical and psychosocial health and there is notably little work that has been done with the Chinese population. A cross-sectional and correlational design was used. A convenience sample of 70 family carers, who were the main carers of stroke patients at home, during the first three months poststroke was recruited to complete a self-report questionnaire. Significant correlations were found between the family carers' global perceived problem-solving abilities and higher level of depressive symptoms (r = 0.35, P = 0.01) and poorer perceived health (r = 0.50, P = 0.01) as measured using the Center for Epidemiological Studies - Depression Scale and General Health Questionnaire. Among the three subscales of the Problem-Solving Inventory, problem-solving confidence showed the highest correlation with these variables. The functional ability of the stroke patients as measured using the Modified Barthel Index (MBI) was not associated with any variables. Findings of this study suggest that perception of confidence is a key factor in appraisal of problem-solving among Chinese family carers, which raises questions for future research about the impact of cultural influences on designing and measuring interventions. The study has implications for nursing and health care practice and for developing interventions targeted at building self-confidence among Chinese carers.


In situ probing of the crystallization kinetics of rr-P3HT on single layer graphene as a function of temperature.
We studied the molecular packing and crystallization of a highly regio-regular semiconducting polymer poly(3-hexylthiophene) (P3HT) on both single layer graphene and silicon as a function of temperature, during cooling from the melt. The onset of crystallization, crystallites' size, orientation, and kinetics of formation were measured in situ by synchrotron grazing incidence X-ray diffraction (GIXD) during cooling and revealed a very different crystallization process on each surface. A favored crystalline orientation with out of plane π-π stacking formed at a temperature of 200 °C on graphene, whereas the first crystallites formed with an edge-on orientation at 185 °C on silicon. The crystallization of face-on lamellae revealed two surprising effects during cooling: (a) a constant low value of the π-π spacing below 60 °C; and (b) a reduction by half in the coherence length of face-on lamellae from 100 to 30 °C, which corresponded with the weakening of the 2nd or 3rd order of the in-plane (k00) diffraction peak. The final ratio of face-on to edge-on orientations was 40% on graphene, and 2% on silicon, revealing the very different crystallization mechanisms. These results provide a better understanding of how surfaces with different chemistries and intermolecular interactions with the polythiophene polymer chains lead to different crystallization processes and crystallites orientations for specific electronic applications.


Novel role of gp91(phox)-containing NAD(P)H oxidase in vascular endothelial growth factor-induced signaling and angiogenesis.
Vascular endothelial growth factor (VEGF) induces angiogenesis by stimulating endothelial cell proliferation and migration, primarily through the receptor tyrosine kinase VEGF receptor2 (Flk1/KDR). Reactive oxygen species (ROS) derived from NAD(P)H oxidase are critically important in many aspects of vascular cell regulation, and both the small GTPase Rac1 and gp91(phox) are critical components of the endothelial NAD(P)H oxidase complex. A role of NAD(P)H oxidase in VEGF-induced angiogenesis, however, has not been defined. In the present study, electron spin resonance spectroscopy is utilized to demonstrate that VEGF stimulates O2*- production, which is inhibited by the NAD(P)H oxidase inhibitor, diphenylene iodonium, as well as by overexpression of dominant-negative Rac1 (N17Rac1) and transfection of gp91(phox) antisense oligonucleotides in human umbilical vein endothelial cells (ECs). Antioxidants, including N-acetylcysteine (NAC), various NAD(P)H oxidase inhibitors, and N17Rac1 significantly attenuate not only VEGF-induced KDR tyrosine phosphorylation but also proliferation and migration of ECs. Importantly, these effects of VEGF are dramatically inhibited in cells transfected with gp91(phox) antisense oligonucleotides. By contrast, ROS are not involved in mediating these effects of sphingosine 1-phosphate (S1P) on ECs. Sponge implant assays demonstrate that VEGF-, but not S1P-, induced angiogenesis is significantly reduced in wild-type mice treated with NAC and in gp91(phox-/-) mice, suggesting that ROS derived from gp91(phox)-containing NAD(P)H oxidase play an important role in angiogenesis in vivo. These studies indicate that VEGF-induced endothelial cell signaling and angiogenesis is tightly controlled by the reduction/oxidation environment at the level of VEGF receptor and provide novel insights into the NAD(P)H oxidase as a potential therapeutic target for angiogenesis-dependent diseases.


Rebates and spreads: pharmacy benefit management practices and corporate citizenship.
How ought we determine whether businesses in the health care sector profit fairly? One class of companies in the health care sector, pharmacy benefit managers (PBMs), deserves special consideration. We describe two specific revenue-generating practices--rebates and spread pricing--that account significantly for PBMs' profits but have been neglected in the bioethics and health policy literature as important sources of fiscal waste in our current health care system. We offer analyses of two common cases, consider employers' and employees' vulnerabilities, explore normative assumptions about how markets function, and raise questions about transparency in contract agreements between PBMs and employers. We consider ethical dimensions of PBMs' corporate citizenship in the health care sector and suggest how employers can negotiate more effectively with PBMs.


DNA-templated plasmonic Ag/AgCl nanostructures for molecular selective photocatalysis and photocatalytic inactivation of cancer cells.
Silver halide (AgX, X = Cl, Br, I)-based materials represent an emerging class of heterogeneous photocatalysts. Despite progress in the synthesis of carrier-separated AgX-based photocatalysts, a number of issues remain unaddressed, including complicated synthesis, unfavorably large size and therefore poor photocatalytic performance of the resultant structures. Here we show the one-step DNA-programmable synthesis of Ag/AgCl nanostructures that takes only approximately 1 min for photocatalytic application. The optimal DNA-encapsulated structures show DNA sequence-specific sizes down to less than 40 nm with a Ag/AgCl composition ratio of 2 : 1, affording a vastly increased surface area and higher photocatalytic activity than any Ag/AgX nanostructures reported previously by over two orders of magnitude. From a physical standpoint, importantly, the plasmonic nanostructured silver in Ag/AgCl accelerates the photocatalytic reaction in terms of fast electron injection to AgCl, leading to enhanced hole-electron separation and high-performance photocatalysis under visible light. To test the effect of DNA encapsulation on the Ag/AgCl nanostructures, both positively and negatively charged organic compounds serve as the model pollutants to assess their photocatalytic selectivity. Our results show that the photodecomposition of the positively charged compounds obeys a first-order rate law, whereas the negatively charged compound is decomposed with zero-order kinetics. This comparison offers a mechanistic insight into reaction kinetics on the DNA-encapsulated photocatalyst. We further find that the DNA-encapsulated Ag/AgCl photocatalysts are robust and can be recycled. To extend the applicability of the Ag/AgCl nanostructures, their use in the efficient photocatalytic inactivation of cancer cells is also demonstrated for the first time, opening up a new avenue to daylight-based theranostics.


Genome-wide mapping of DNase hypersensitive sites using massively parallel signature sequencing (MPSS).
A major goal in genomics is to understand how genes are regulated in different tissues, stages of development, diseases, and species. Mapping DNase I hypersensitive (HS) sites within nuclear chromatin is a powerful and well-established method of identifying many different types of regulatory elements, but in the past it has been limited to analysis of single loci. We have recently described a protocol to generate a genome-wide library of DNase HS sites. Here, we report high-throughput analysis, using massively parallel signature sequencing (MPSS), of 230,000 tags from a DNase library generated from quiescent human CD4+ T cells. Of the tags that uniquely map to the genome, we identified 14,190 clusters of sequences that group within close proximity to each other. By using a real-time PCR strategy, we determined that the majority of these clusters represent valid DNase HS sites. Approximately 80% of these DNase HS sites uniquely map within one or more annotated regions of the genome believed to contain regulatory elements, including regions 2 kb upstream of genes, CpG islands, and highly conserved sequences. Most DNase HS sites identified in CD4+ T cells are also HS in CD8+ T cells, B cells, hepatocytes, human umbilical vein endothelial cells (HUVECs), and HeLa cells. However, approximately 10% of the DNase HS sites are lymphocyte specific, indicating that this procedure can identify gene regulatory elements that control cell type specificity. This strategy, which can be applied to any cell line or tissue, will enable a better understanding of how chromatin structure dictates cell function and fate.


Can peripheral psychophysiological markers predict response to exposure-based cognitive behavioral therapy in youth with severely impairing irritability? A study protocol.
Irritability, an increased proneness to anger, is a primary reason youth present for psychiatric care. While initial evidence supports the efficacy of exposure-based cognitive behavioral therapy (CBT) for youth with clinically impairing irritability, treatment mechanisms remain unclear. Here, we propose to measure peripheral psychophysiological indicators of arousal-heart rate (HR)/electrodermal activity (EDA)-and regulation-heart rate variability (HRV)-during exposures to anger-inducing stimuli as potential predictors of treatment efficacy. The objective of this study is to evaluate whether in-situ biosensing data provides peripheral physiological indicators of in-session response to exposures. Blood volume pulse (BVP; from which HR and HRV canl be derived) and EDA will be collected ambulatorily using the Empatica EmbracePlus from 40 youth (all genders; ages 8-17) undergoing six in-person exposure treatment sessions, as part of a multiple-baseline trial of exposure-based CBT for clinically impairing irritability. Clinical ratings of irritability will be conducted at baseline, weekly throughout treatment, and at 3-month and 6-month follow-ups via the Clinical Global Impressions Scale (CGI) and the Affective Reactivity Index (ARI; clinician-, parent-, and child-report). Multilevel modeling will be used to assess within- and between-person changes in physiological arousal and regulation throughout exposure-based CBT and to determine whether individual differences are predictive of treatment response. This study protocol leverages a wearable biosensor (Empatica) to continuously record HR/HRV (derived from BVP) and EDA during in-person exposure sessions for youth with clinically impairing irritability. Here, the goal is to identify changes in physiological arousal (EDA, HR) and regulation (HRV) over the course of treatment in tandem with changes in clinical symptoms. The participants in this study come from an overarching clinical trial (trial registration numbers: NCT02531893 first registered on 8/25/2015; last updated on 8/25/2023). The research project and all related materials were submitted and approved by the appropriate Institutional Review Board of the National Institute of Mental Health (NIMH).


Effect of chemical interaction between oleic acid and L-Arginine on oral perception, as a function of polymorphisms of CD36 and OBPIIa and genetic ability to taste 6-n-propylthiouracil.
Oral sensitivity to fats varies in individuals influencing nutritional status and health. Variations in oleic acid perception are associated with CD36 and odorant binding protein (OBPIIa) polymorphisms, and 6-n-propylthiouracil (PROP) sensitivity, which is mediated by TAS2R38 receptor. L-Arginine (L-Arg) supplementation was shown to modify the perception of the five taste qualities. Here we analyzed the effect of three concentrations (5, 10, 15 mmol/L) of L-Arg on oral perception of oleic acid in forty-six subjects classified for PROP taster status and genotyped for TAS2R38, CD36 and OBPIIa polymorphisms. L-Arg supplementation was effective in increasing the perceived intensity of oleic acid in most subjects. The lowest concentration was the most effective, especially in PROP non-tasters or medium tasters, and in subjects with at least an allele A in CD36 and OBPIIa loci. Density Functional Theory (DFT) calculations were exploited to characterize the chemical interaction between L-Arg and oleic acid, showing that a stable 1:1 oleate·ArgH+ adduct can be formed, stabilized by a pair of hydrogen bonds. Results indicate that L-Arg, acting as a 'carrier' of fatty acids in saliva, can selectively modify taste response, and suggest that it may to be used in personalized dietetic strategies to optimize eating behaviors and health.


Cortical Folding of the Primate Brain: An Interdisciplinary Examination of the Genetic Architecture, Modularity, and Evolvability of a Significant Neurological Trait in Pedigreed Baboons (Genus Papio).
Folding of the primate brain cortex allows for improved neural processing power by increasing cortical surface area for the allocation of neurons. The arrangement of folds (sulci) and ridges (gyri) across the cerebral cortex is thought to reflect the underlying neural network. Gyrification, an adaptive trait with a unique evolutionary history, is affected by genetic factors different from those affecting brain volume. Using a large pedigreed population of ∼1000 Papio baboons, we address critical questions about the genetic architecture of primate brain folding, the interplay between genetics, brain anatomy, development, patterns of cortical-cortical connectivity, and gyrification's potential for future evolution. Through Mantel testing and cluster analyses, we find that the baboon cortex is quite evolvable, with high integration between the genotype and phenotype. We further find significantly similar partitioning of variation between cortical development, anatomy, and connectivity, supporting the predictions of tension-based models for sulcal development. We identify a significant, moderate degree of genetic control over variation in sulcal length, with gyrus-shape features being more susceptible to environmental effects. Finally, through QTL mapping, we identify novel chromosomal regions affecting variation in brain folding. The most significant QTL contain compelling candidate genes, including gene clusters associated with Williams and Down syndromes. The QTL distribution suggests a complex genetic architecture for gyrification with both polygeny and pleiotropy. Our results provide a solid preliminary characterization of the genetic basis of primate brain folding, a unique and biomedically relevant phenotype with significant implications in primate brain evolution.


Macroscopic fibers and ribbons of oriented carbon nanotubes.
A simple method was used to assemble single-walled carbon nanotubes into indefinitely long ribbons and fibers. The processing consists of dispersing the nanotubes in surfactant solutions, recondensing the nanotubes in the flow of a polymer solution to form a nanotube mesh, and then collating this mesh to a nanotube fiber. Flow-induced alignment may lead to a preferential orientation of the nanotubes in the mesh that has the form of a ribbon. Unlike classical carbon fibers, the nanotube fibers can be strongly bent without breaking. Their obtained elastic modulus is 10 times higher than the modulus of high-quality bucky paper.


Contamination of fresh and ensiled maize by multiple penicillium mycotoxins.
Toxins produced by Penicillium species are reported in maize silage and have been associated with health problems in cattle. Our objectives were to evaluate the prevalence and dynamics of patulin (PAT), mycophenolic acid (MPA), cyclopiazonic acid (CPA), and roquefortine C (ROC) in fresh and ensiled maize. To achieve these objectives we developed a high-performance liquid chromatography method coupled with mass spectrometry to detect all four toxins simultaneously in silage. In addition we collected weather data, information on agronomic practices, and silage fermentation characteristics for each study site. Silage was collected at harvest and after ensiling in 2001 and 2002 from 30 Pennsylvania dairies. The average concentration of toxins (range in parentheses) was: PAT 0.08 microg/g (0.01 to 1.21), MPA 0.16 microg/g (0.02 to 1.30), CPA 0.12 microg/g (0.02 to 1.43), and ROC 0.38 microg/g (0.01 to 5.71). ROC was the most frequently detected toxin (60%), followed by MPA (42%), CPA (37%), and PAT (23%). Of 120 samples tested, 15% contained no detectible levels of toxin, 25% were contaminated with one toxin, 32% with two, 18% with three, and 10% with all four toxins. All four mycotoxins were found in freshly harvested material, contradicting the belief that Penicillium toxin formation occurs exclusively during storage. We observed that weather conditions during specific growth stages of the crop affected the final concentration of toxins in freshly harvested maize. In ensiled material, PAT levels were affected by concentrations of propionic and isobutyric acids. Based on our data, Penicillium mycotoxins can form while the crop is in the field and after ensiling, suggesting that preventative measures should begin prior to ensiling.


Identification of mycorrhiza-regulated genes with arbuscule development-related expression profile.
Suppressive subtractive hybridisation was applied to the analysis of late stage arbuscular mycorrhizal development in pea. 96 cDNA clones were amplified and 81, which carried fragments more than 200 nt in size, were sequence analysed. Among 67 unique fragments, 10 showed no homology and 10 were similar to sequences with unknown function. RNA accumulation of the corresponding 67 genes was analysed by hybridisation of macro-arrays. The cDNAs used as probes were derived from roots of wild type and late mutant pea genotypes, inoculated or not with the AM fungus Glomus mosseae. After calibration, a more than 2.5-fold mycorrhiza-induced RNA accumulation was detected in two independent experiments in the wild type for 25 genes, 22 of which seemed to be induced specifically during late stage AM development. Differential expression for 7 genes was confirmed by RT-PCR using RNA from mycorrhiza and from controls of a different pea cultivar. In order to confirm arbuscule-related expression, the Medicago truncatula EST data base was screened for homologous sequences with putative mycorrhiza-related expression and among a number of sequences with significant similarities, a family of trypsin inhibitor genes could be identified. Mycorrhiza-induced RNA accumulation was verified for five members by real-time PCR and arbuscule-related activation of the promoter could be shown in transgenic roots for one of the genes, MtTi 1.


Hemodilution increases cerebral blood flow in acute ischemic stroke.
We measured cerebral blood flow in 10 consecutive, but selected, patients with acute ischemic stroke (less than 48 hours after onset) before and after hemodilution. Cerebral blood flow was measured by xenon-133 inhalation and emission tomography, and only patients with focal hypoperfusion in clinically relevant areas were included. Hemodilution was done according to the hematocrit level: for a hematocrit greater than or equal to 42%, 500 ml whole blood was drawn and replaced by the same volume of dextran 40; for a hematocrit between 37% and 42%, only 250 ml whole blood was drawn and replaced by 500 cc of dextran 40. Mean hematocrit was reduced by 16%, from 46 +/- 5% (SD) to 39 +/- 5% (SD) (p less than 0.001). Cerebral blood flow increased in both hemispheres by an average of 20.9% (p less than 0.001). Regional cerebral blood flow increased in the ischemic areas in all cases, on an average of 21.4 +/- 12.0% (SD) (p less than 0.001). In three patients, a significant redistribution of flow in favor of the hypoperfused areas was observed, and in six patients, the fractional cerebral blood flow increase in the hypoperfused areas was of the same magnitude as in the remainder of the brain. In the last patient, cerebral blood flow increased relatively less in the ischemic areas. Our findings show that cerebral blood flow increases in the ischemic areas after hemodilution therapy in stroke patients. The marked regional cerebral blood flow increase seen in some patients could imply an improved oxygen delivery to the ischemic tissue.


Cubosomes for topical delivery of the antimicrobial peptide LL-37.
In this study, the use of cubosomes for topical delivery of the antimicrobial peptide (AMP) LL-37 was investigated. Topical delivery of AMPs is of great interest for treatment of skin infections caused by bacteria, such as Staphylococcus aureus. AMP containing cubosomes were produced by three different preparation protocols and compared: (i) pre-loading, where LL-37 was incorporated into a liquid crystalline gel, which thereafter was dispersed into nanoparticles, (ii) post-loading, where LL-37 was let to adsorb onto pre-formed cubosomes, and (iii) hydrotrope-loading, where LL-37 was incorporated during the spontaneously formed cubosomes in an ethanol/glycerol monooleate mixture. Particle size and size distribution were analyzed using dynamic light scattering (DLS), liquid crystalline structure by small angle x-ray scattering (SAXS) and release of LL-37 by a fluorescamine assay. Proteolytic protection of LL-37 as well as bactericidal effect after enzyme exposure was investigated. The skin irritation potential of cubosomes was examined by an in vitro epidermis model. Finally, the bacterial killing property of the cubosomes was examined by an ex vivo pig skin wound infection model with Staphylococcus aureus. Data showed that a high loading of LL-37 induced formation of vesicles in case of cubosomes prepared by sonication (pre-loading). No release of LL-37 was observed from the cubosomes, indicating strong association of the peptide to the particles. Proteolysis studies showed that LL-37 was fully protected against enzymatic attacks while associated with the cubosomes, also denoting strong association of the peptide to the particles. As a consequence, bactericidal effect after enzyme exposure remained, compared to pure LL-37 which was subjected to proteolysis. No skin irritation potential of the cubosomes was found, thus enabling for topical administration. The ex vivo wound infection model showed that LL-37 in pre-loaded cubosomes killed bacteria most efficient.


Nucleation, fiber growth and melting, and domain formation and structure in sickle cell hemoglobin gels.
Pathogenesis in sickle cell disease depends on polymerization and gelation of deoxyhemoglobin S. Under the double nucleation model, polymerization is initiated by homogeneous nucleation, followed by heterogeneous nucleation on pre-existing fibers. Fibers grow by non-cooperative addition of hemoglobin. The model derives from macroscopic results rather than direct observation of individual events. We observe individual events and structures by differential interference contrast (DIC) microscopy to show consistency with the model, to define structure and development of gel domains and their relation to kinetics, and to demonstrate the mechanism of fiber melting. Kinetics were controlled by producing deoxyhemoglobin by photolysis of CO hemoglobin under DIC observation. The first visible polymers appeared randomly and were usually linear aggregates less than 1 micron long, consistent with homogeneous nucleation and immediate post-nucleation aggregates. Aggregates then branched extensively, consistent with heterogeneous nucleation. This branching of new fibers was also induced at countable rates on isolated single fibers. Branching and fiber growth rapidly produced dense domains. Changes in photolytic intensity altered domain growth rates and domain structure. At low intensity and slow growth, fibers grew radially without branching. Domains lacked cross-links and polymer density was low. High intensity produced faster growth, much heterogeneous nucleation and highly cross-linked, dense, domains. At still higher intensity, homogeneous nucleation was very rapid, producing many small domains. These results show a hierarchy of processes: as deoxyhemoglobin concentration increases, growth occurs without observable nucleations, and then heterogeneous and finally homogeneous nucleation become dominant. This is consistent with the double nucleation model under which the concentration dependence of growth is low, and that of heterogeneous and homogeneous nucleation successively higher. Under decreased photolysis, fiber ends melted continuously without fiber breakage; increased photolysis reversed this, producing growth. Isolated fibers melted and grew at both ends. The results are consistent with a fiber melting mechanism that is the reverse of growth.


The countryside information system: A strategic-level decision support system.
The Institute of Terrestrial Ecology (ITE) has monitored ecological change in Great Britain (GB) since 1978. The task has been undertaken using a stratified sampling scheme working with a 1 km square as the sample unit. In more recent years, scientific researchers at ITE have been working closely with the policy-makers of the United Kingdom Department of the Environment. The presentation of information to policy advisors and planners was a component within a large project investigating the ecological consequences of land-use change. A simple PC-based decision support system was developed during the project and subsequently has been expanded to produce a marketable product. The system, called the Countryside Information System (CIS), presents and links information at national, regional and thematic levels along with qualifying data describing accuracy and appropriateness of use (i.e., metadata). An integral part of the CIS is the ITE Land Classification, which divides GB into 32 environmental land classes; all 250 000 squares have been classified. The classification allows sampled data to be presented and, as the co-ordinate system is widely used in GB, it allows census datasets to be linked and compared. CIS has been described as a Geographical Information System, but the classification, data held within the system, and the use of metadata to assist in interpretation of results make the system much more decision-support oriented. Indeed, government departments have been involved in directing the development and are now starting to use the system to answer parliamentary questions and formulate, assess and monitor environmental policy. The CIS is an open system, running on a standard PC in Microsoft Windows. Tools for loading and editing new datasets (both sample and census) are incorporated in the suite of programs. The Windows environment and users comments during development have produced a system with an intuitive feel, removing some of the overhead of acquiring specialised technical skills before being able to operate a system. This paper describes the CIS and presents examples of its applications.


Association Between Lower Preoperative Cognition With Intraoperative Electroencephalographic Features Consistent With Deep States of Anesthesia in Older Patients: An Observational Cohort Study.
Patients with low cognitive performance are thought to have a higher risk of postoperative neurocognitive disorders. Here we analyzed the relationship between preoperative cognition and anesthesia-induced brain dynamics. We hypothesized that patients with low cognitive performance would be more sensitive to anesthetics and would show differences in electroencephalogram (EEG) activity consistent with a brain anesthesia overdose. This is a retrospective analysis from a previously reported observational study. We evaluated cognitive performance using the Montreal cognitive assessment (MoCA) test. All patients received general anesthesia maintained with sevoflurane or desflurane during elective major abdominal surgery. We analyzed the EEG using spectral, coherence, and phase-amplitude modulation analyses. Patients were separated into a low MoCA group (<26 points, n = 12) and a high MoCA group (n = 23). There were no differences in baseline EEG, nor end-tidal age-corrected minimum alveolar concentration (MACage). However, under anesthesia, the low MoCA group had lower α-β power (high MoCA: 2.9 [interquartile range {IQR}: 0.6-5.8 dB] versus low MoCA: -1.2 [IQR: -2.1 to 0.6 dB], difference 4.1 [1.0-5.7]) and a lower α peak frequency (high MoCA: 9.0 [IQR: 8.3-9.8 Hz] versus low MoCA: 7.5 [IQR: 6.3-9.0 Hz], difference 1.5 [0-2.3]) compared to the high MoCA group. The low MoCA group also had a lower α band coherence and a stronger peak-max phase-amplitude coupling (PAC). Finally, patients in the low MoCA group had longer emergence times (high MoCA 663 ± 345 seconds versus low MoCA: 960 ± 352 seconds, difference 297 [15-578]). Multiple linear regression shows up that both age and MoCA scores are independently associated with intraoperative α-β power. All these EEG features, together with a prolonged emergence time, are consistent with the possibility that older patients with low cognitive performance are receiving a brain anesthesia overdose compare to cognitive normal patients.


[Phase I study of paclitaxel].
Paclitaxel, a novel antimicrotubule agent that enhances tubulin polymerization and microtubule stability, was administered as a 24-hour infusion in a phase I study. Twelve patients received 32 courses at 50, 100, 150, and 200 mg/m2. A premedication regimen of dexamethasone, diphenhydramine, and ranitidine was used to prevent the acute hypersensitivity reactions (HSRs). The dose-limiting factor was leukopenia (granulocytopenia) associated with Grade 4 infection. The maximum tolerated dose was 200 mg/m2. Other non-hematological effects included peripheral neuropathy, myalgia, alopecia, and elevations of transaminase and alkaline phosphatase. Severe HSRs were not observed. The paclitaxel plasma concentration declined with a half-life of 10.0 to 24.9 hours. Excretion into urine within 72 hours was in the range of 7.28 to 11.34% of paclitaxel dosage. Two patients with breast cancer at the 200 mg/m2 dose level had partial responses. The recommended dose of paclitaxel for phase II study, when administered as a 24-hour infusion, is considered to be 150 mg/m2 every 3 weeks.


'I know it's wrong, but...': a qualitative investigation of low-income parents' feelings of guilt about their child-feeding practices.
In the developed world, child overweight and obesity rates are highest among the disadvantaged. This has resulted in calls for more research with low socio-economic families to better understand their experiences with disadvantage and how they might lead to poorer weight outcomes. The present study, conducted in Australia, adopted a qualitative approach to investigate the factors affecting low socio-economic parents' child-feeding practices. Methods used to collect data were introspections, interviews and focus groups. In total, 37 parents of overweight or obese children aged between 5 and 9 years took part in the 6-month study. Guilt emerged as an emotion that parents regularly experienced when allowing their children to consume too much food or foods high in fat, salt and/or sugar. Parents attributed their guilt-inducing child-feeding practices to both external and internal factors. Time scarcity and cost were factors that were primarily characterized by an external locus of control. The factors characterized by an internal locus of control were fear of their children experiencing hunger, the perceived need to secure their children's affection through the provision of treat foods, perceptions of their ability to balance their children's diets across eating situations and perceived laziness. Recommendations are provided for addressing guilt-inducing child-feeding practices.


Comparison of gene expression profile of epiretinal membranes obtained from eyes with proliferative vitreoretinopathy to that of secondary epiretinal membranes.
Proliferative vitreoretinopathy (PVR) is a destructive complication of retinal detachment and vitreoretinal surgery which can lead to severe vision reduction by tractional retinal detachments. The purpose of this study was to determine the gene expression profile of epiretinal membranes (ERMs) associated with a PVR (PVR-ERM) and to compare it to the expression profile of less-aggressive secondary ERMs. A PCR-amplified complementary DNA (cDNA) library was constructed using the RNAs isolated from ERMs obtained during vitrectomy. The sequence from the 5' end was obtained for randomly selected clones and used to generate expressed sequence tags (ESTs). We obtained 1116 nonredundant clusters representing individual genes expressed in PVR-ERMs, and 799 clusters representing the genes expressed in secondary ERMs. The transcriptome of the PVR-ERMs was subdivided by functional subsets of genes related to metabolism, cell adhesion, cytoskeleton, signaling, and other functions, by FatiGo analysis. The genes highly expressed in PVR-ERMs were compared to those expressed in the secondary ERMs, and these were subdivided by cell adhesion, proliferation, and other functions. Querying 10 cell adhesion-related genes against the STRING database yielded 70 possible physical relationships to other genes/proteins, which included an additional 60 genes that were not detected in the PVR-ERM library. Of these, soluble CD44 and soluble vascular cellular adhesion molecule-1 were significantly increased in the vitreous of patients with PVR. Our results support an earlier hypothesis that a PVR-ERM, even from genomic points of view, is an aberrant form of wound healing response. Genes preferentially expressed in PVR-ERMs may play an important role in the progression of PVR and could be served as therapeutic targets.


Long-term predictable nipple projection following reconstruction.
The creation of the nipple-areola complex is often the final step in the surgical treatment of breast cancer patients, and it consequently has important symbolic and aesthetic implications. Patient expectations and the need for symmetry make nipple projection a crucial aesthetic determinant of nipple reconstruction. We hypothesize that long-term nipple projection and shape can be achieved in a predictable fashion using the modified star dermal fat flap technique. Prospectively, 93 nipples were reconstructed by a single surgeon using a modified star dermal fat flap technique in 44 implant and 49 TRAM flap breast reconstructions. Flap dimensions (base diameter and flap length) were designed according to patient desire or to the base diameter and projection of the opposite breast nipple. A standardized, 3-month postoperative care regimen was observed in all patients. Nipple projection was assessed by the same observer at each follow-up examination. The average length of follow-up was 730 days (745 for TRAM reconstructions and 713 for implants). Consistently, an average of 41 percent of the intraoperative projection remained intact in both groups at final evaluation (SD 12 percent). The total flap length was strongly predictive of intraoperative and long-term projection (r = 0.64 and 0.86, p < 0.0001). Flap lengths ranged from 5.5 to 9.0 cm, and in a linear correlation, resulted in intraoperative projection of 1.0 to 2.1 cm, respectively, and long-term projection of 0.4 to 0.83 cm, respectively. Based on the linear relationship, every 1-cm increase in flap length could be expected to result in a 0.16-cm increase in projection. When controlled for flap length and intraoperative projection, there was no difference between TRAM and implant nipple reconstruction in predicting postoperative nipple projection. Intraoperative planning and execution are critical to achieve predictable nipple shape, size, and projection. The dimensions of the star dermal fat flap can be strategically modified to allow the surgeon predictable projection with a consistent 41-percent preservation of intraoperative nipple projection in both TRAM and implant patients at 2 years.


Association of Donors With US Public Health Service Risk Criteria and Outcomes After Adult vs Pediatric Cardiac Transplant.
The United Network for Organ Sharing (UNOS) evaluates donor risk for acute transmission of HIV, hepatitis B, or hepatitis C based on US Public Health Services (PHS)-specific criteria. However, recent data regarding use and outcomes of those donors with PHS risk criteria among pediatric and adult heart transplant recipients are lacking. To compare use and outcomes of graft from donors with PHS risk criteria vs those with a standard-risk donor (SRD) in children vs adults in a contemporary cohort. This cohort was a nationwide analysis of heart transplants in the US that used data from the UNOS database. Participants were children (<18 years old) and adults (≥18 years old) who received a heart transplant from January 1, 2010, to December 31, 2021. UNOS-defined donor risk status. Trend analysis compared changes in PHS risk criteria use among children and adults. Patient survival was analyzed using Kaplan-Meier curves with log rank and Cox proportional hazards to compare PHS risk-criteria outcomes vs SRD-criteria outcomes in children and adult heart transplant recipients. Additional analysis was performed among adults who received a PHS-risk criteria graft that was previously declined for pediatric recipients. Of 5115 pediatric transplant recipients (donor without PHS risk median [IQR] age, 5 [0-13] years and donor with PHS risk median [IQR] age, 8 [0-14] years) and 30 289 adult heart transplant recipients (donor without PHS risk median [IQR] age, 56 [46-63] years and donor with PHS risk median [IQR] age, 57 [47-63] years), PHS risk criteria comprised 8% in children vs 25% in adults. PHS criteria are being increasingly used over the past decade with the proportion of recipients transplanted with PHS risk-criteria donors being approximately 3 times greater among adult recipients than children recipients. Pediatric recipients of a PHS risk-criteria donor had greater pretransplant ventilatory support, whereas adult recipients of a PHS risk-criteria donor had greater pretransplant extracorporeal membrane oxygenation use. Patient survival was similar between pediatric recipients of PHS risk-criteria grafts vs SRD-criteria grafts and slightly higher among adult recipients of PHS risk-criteria grafts vs SRD-criteria grafts. The 1778 adult recipients who received a PHS criteria-risk donor that was previously declined for pediatric recipients had similar patient survival recipients compared with SRD-criteria donors (HR, 0.92; 95% CI, 0.81-1.03; P = .18). In the current era, a 3-fold greater proportion of adult recipients receive a PHS risk-criteria graft compared with children despite similar posttransplant patient survival. The ongoing organ donor shortage underscores the need for consideration of PHS risk criteria where these donors remain underused.


Power changes how the brain responds to others.
Power dynamics are a ubiquitous feature of human social life, yet little is known about how power is implemented in the brain. Motor resonance is the activation of similar brain networks when acting and when watching someone else act, and is thought to be implemented, in part, by the human mirror system. We investigated the effects of power on motor resonance during an action observation task. Separate groups of participants underwent a high-, neutral, or low-power induction priming procedure, prior to observing the actions of another person. During observation, motor resonance was determined with transcranial magnetic stimulation (TMS) via measures of motor cortical output. High-power participants demonstrated lower levels of resonance than low-power participants, suggesting reduced mirroring of other people in those with power. These differences suggest that decreased motor resonance to others' actions might be one of the neural mechanisms underlying power-induced asymmetries in processing our social interaction partners.


Vertebrate innovations.
Vertebrate innovations include neural crest cells and their derivatives, neurogenic placodes, an elaborate segmented brain, endoskeleton, and an increase in the number of genes in the genome. Comparative molecular and developmental data give new insights into the evolutionary origins of these characteristics and the complexity of the vertebrate body.


87Sr solid-state NMR as a structurally sensitive tool for the investigation of materials: antiosteoporotic pharmaceuticals and bioactive glasses.
Strontium is an element of fundamental importance in biomedical science. Indeed, it has been demonstrated that Sr(2+) ions can promote bone growth and inhibit bone resorption. Thus, the oral administration of Sr-containing medications has been used clinically to prevent osteoporosis, and Sr-containing biomaterials have been developed for implant and tissue engineering applications. The bioavailability of strontium metal cations in the body and their kinetics of release from materials will depend on their local environment. It is thus crucial to be able to characterize, in detail, strontium environments in disordered phases such as bioactive glasses, to understand their structure and rationalize their properties. In this paper, we demonstrate that (87)Sr NMR spectroscopy can serve as a valuable tool of investigation. First, the implementation of high-sensitivity (87)Sr solid-state NMR experiments is presented using (87)Sr-labeled strontium malonate (with DFS (double field sweep), QCPMG (quadrupolar Carr-Purcell-Meiboom-Gill), and WURST (wideband, uniform rate, and smooth truncation) excitation). Then, it is shown that GIPAW DFT (gauge including projector augmented wave density functional theory) calculations can accurately compute (87)Sr NMR parameters. Last and most importantly, (87)Sr NMR is used for the study of a (Ca,Sr)-silicate bioactive glass of limited Sr content (only ~9 wt %). The spectrum is interpreted using structural models of the glass, which are generated through molecular dynamics (MD) simulations and relaxed by DFT, before performing GIPAW calculations of (87)Sr NMR parameters. Finally, changes in the (87)Sr NMR spectrum after immersion of the glass in simulated body fluid (SBF) are reported and discussed.


Sulfonated red and far-red rhodamines to visualize SNAP- and Halo-tagged cell surface proteins.
The (in)ability to permeate membranes is a key feature of chemical biology probes that defines their suitability for specific applications. Here we report sulfonated rhodamines that endow xanthene dyes with cellular impermeability for analysis of surface proteins. We fuse charged sulfonates to red and far-red dyes to obtain Sulfo549 and Sulfo646, respectively, and further link these to benzylguanine and choloralkane substrates for SNAP-tag and Halo-tag labelling. Sulfonated rhodamine-conjugated fluorophores maintain desirable photophysical properties, such as brightness and photostability. While transfected cells with a nuclear localized SNAP-tag remain unlabelled, extracellular exposed tags can be cleanly visualized. By multiplexing with a permeable rhodamine, we are able to differentiate extra- and intracellular SNAP- and Halo-tags, including those installed on the glucagon-like peptide-1 receptor, a prototypical class B G protein-coupled receptor. Sulfo549 and Sulfo646 also labelled transfected neurons derived from induced pluripotent stem cells (iPSCs), allowing STED nanoscopy of the axonal membrane. Together, this work provides a new avenue for rendering dyes impermeable for exclusive extracellular visualization via self-labelling protein tags. We anticipate that Sulfo549, Sulfo646 and their congeners will be useful for a number of cell biology applications where labelling of intracellular sites interferes with accurate surface protein analysis.


Multi-component assessment of chronic obstructive pulmonary disease: an evaluation of the ADO and DOSE indices and the global obstructive lung disease categories in international primary care data sets.
Suitable tools for assessing the severity of chronic obstructive pulmonary disease (COPD) include multi-component indices and the global initiative for chronic obstructive lung disease (GOLD) categories. The aim of this study was to evaluate the dyspnoea, obstruction, smoking, exacerbation (DOSE) and the age, dyspnoea, obstruction (ADO) indices and GOLD categories as measures of current health status and future outcomes in COPD patients. This was an observational cohort study comprising 5,114 primary care COPD patients across three databases from UK, Sweden and Holland. The associations of DOSE and ADO indices with (i) health status using the Clinical COPD Questionnaire (CCQ) and St George's Respiratory Questionnaire (SGRQ) and COPD Assessment test (CAT) and with (ii) current and future exacerbations, admissions and mortality were assessed in GOLD categories and DOSE and ADO indices. DOSE and ADO indices were significant predictors of future exacerbations: incident rate ratio was 1.52 (95% confidence intervals 1.46-1.57) for DOSE, 1.16 (1.12-1.20) for ADO index and 1.50 (1.33-1.68) and 1.23 (1.10-1.39), respectively, for hospitalisations. Negative binomial regression showed that the DOSE index was a better predictor of future admissions than were its component items. The hazard ratios for mortality were generally higher for ADO index groups than for DOSE index groups. The GOLD categories produced widely differing assessments for future exacerbation risk or for hospitalisation depending on the methods used to calculate them. None of the assessment systems were excellent at predicting future risk in COPD; the DOSE index appears better than the ADO index for predicting many outcomes, but not mortality. The GOLD categories predict future risk inconsistently. The DOSE index and the GOLD categories using exacerbation frequency may be used to identify those at high risk for exacerbations and admissions.


Emergence of a Novel Salmonella enterica Serotype Reading Clonal Group Is Linked to Its Expansion in Commercial Turkey Production, Resulting in Unanticipated Human Illness in North America.
Two separate human outbreaks of Salmonella enterica serotype Reading occurred between 2017 and 2019 in the United States and Canada, and both outbreaks were linked to the consumption of raw turkey products. In this study, a comprehensive genomic investigation was conducted to reconstruct the evolutionary history of S. Reading from turkeys and to determine the genomic context of outbreaks involving this infrequently isolated Salmonella serotype. A total of 988 isolates of U.S. origin were examined using whole-genome-based approaches, including current and historical isolates from humans, meat, and live food animals. Broadly, isolates clustered into three major clades, with one apparently highly adapted turkey clade. Within the turkey clade, isolates clustered into three subclades, including an "emergent" clade that contained only isolates dated 2016 or later, with many of the isolates from these outbreaks. Genomic differences were identified between emergent and other turkey subclades, suggesting that the apparent success of currently circulating subclades is, in part, attributable to plasmid acquisitions conferring antimicrobial resistance, gain of phage-like sequences with cargo virulence factors, and mutations in systems that may be involved in beta-glucuronidase activity and resistance towards colicins. U.S. and Canadian outbreak isolates were found interspersed throughout the emergent subclade and the other circulating subclade. The emergence of a novel S Reading turkey subclade, coinciding temporally with expansion in commercial turkey production and with U.S. and Canadian human outbreaks, indicates that emergent strains with higher potential for niche success were likely vertically transferred and rapidly disseminated from a common source.IMPORTANCE Increasingly, outbreak investigations involving foodborne pathogens are difficult due to the interconnectedness of food animal production and distribution, and homogeneous nature of industry integration, necessitating high-resolution genomic investigations to determine their basis. Fortunately, surveillance and whole-genome sequencing, combined with the public availability of these data, enable comprehensive queries to determine underlying causes of such outbreaks. Utilizing this pipeline, it was determined that a novel clone of Salmonella Reading has emerged that coincided with increased abundance in raw turkey products and two outbreaks of human illness in North America. The rapid dissemination of this highly adapted and conserved clone indicates that it was likely obtained from a common source and rapidly disseminated across turkey production. Key genomic changes may have contributed to its apparent continued success in commercial turkeys and ability to cause illness in humans.


[Survey of surgical treatment of malignant pleural mesothelioma in Japan].
Surgical results of patients with malignant pleural mesothelioma (MPM) in Japan was surveyed from the results of a questionnaire sent to members of the Japan Lung Cancer Society. and a total of 132 surgical cases of MPM from 1997 to 2002 were analyzed. They consisted of 112 males and 20 females. By histological type, 87 cases had epithelial type, 10 had sarcomatous type, 26 had mixed type and 2 had uncommon type of MPM (the histology of 7 cases was not indicated). As to the surgical mode, extrapleural pneumonectomy (EPP) was performed in 73 cases, and limited surgery such as decortication and tumorectomy was performed in 59 cases. The tumor was potentially completely resected in 83 cases. Postoperative adjuvant therapy was performed in 56 cases. The 1-, 2- and 3-year survival rates of the present cases were 54, 33 and 21%, respectively, and the perioperative mortality rate was 5%. These survival and mortality rates in the present series were almost similar to those of the MPM cases in the previously reported series from 1987 to 1996 by Takagi et al. According to Cox regression analysis, prognostic factors for survival included postoperative adjuvant therapy (p=0.003) and complete resection (p=0.037) significantly, and International Mesothelioma Interest Group (IMIG) stage (p=0.051) and performance status (p=0.086) with a marginal significance, indicating that complete surgical resection of the tumor and perioperative adjuvant therapy could be effective treatment for MPM in Japan. Thus, the development of multimodality therapy including surgical treatment for this disease may be required to improve surgical results of MPM patients.


Navigating complexity to support justice-involved youth with FASD and other neurodevelopmental disabilities: needs and challenges of a regional workforce.
Young people with Fetal Alcohol Spectrum Disorder (FASD) can face significant challenges in their lives, including overrepresentation in the justice system from a young age. Police questioning and court proceedings can be difficult for these young people to navigate. Practice and policy responses are necessary to identify these individuals, provide appropriate support/rehabilitation, and upskill the justice workforce. The aim of this research was to determine the unmet workforce development needs of a regional workforce providing care and support to youth involved with the justice system. Interviews were conducted with 29 participants from 14 organisations to understand the support provided to youth, existence and uptake of referral pathways, and unmet needs. Results revealed a workforce that wants to see improvements to outcomes for young people with FASD and other neurodevelopmental disabilities who enter the youth justice system. However more support is required through training, ongoing funding, and assistance to develop FASD informed work practices. The workforce supporting youth to navigate the justice system requires practical interventions to achieve best practice so that young people with FASD and other neurodevelopmental disabilities receive the support that they need. Following the interviews a model of care tool was developed and piloted in the sector. The tool includes current pathways through the justice system and provides resources to assist staff in achieving best practice care for young people with FASD and other neurodevelopmental disabilities.


Conditions that may affect the results of susceptibility testing of Mycobacterium tuberculosis to pyrazinamide.
Pyrazinamide (PZA) is an important front-line anti-tuberculosis drug that is active only at acid pH. However, acid pH causes significant difficulty for PZA susceptibility testing. A common problem in PZA testing is false resistance caused by large bacterial inocula. This study investigated the relationship of false resistance to numbers of bacilli, pH and other factors that potentially affect susceptibility to PZA. Large inocula (10(7-8) bacilli/ml) of M. tuberculosis H37Ra caused significant increase in medium pH from 5.5 towards neutrality, and thus produced false resistance results. The increase in medium pH was determined to be a function of live bacilli; heat-killed bacilli had little or no effect. Susceptibility to PZA and its active derivative pyrazinoic acid (POA) was comparable on 7H11 agar medium, but POA was less active than PZA in liquid medium containing bovine serum albumin (BSA), suggesting that susceptibility to PZA or POA was reduced in the presence of BSA, because of its neutralising effect on medium pH and significant POA binding. A 3-month-old H37Ra culture was shown to be more susceptible to PZA exposure than a 4-day log-phase culture, suggesting that PZA is more active for non-growing bacilli. Finally, reserpine, an inhibitor of POA efflux pump, increased susceptibility to PZA even near neutral pH 6.8, with an MIC of 400 mg/L compared with 1,000 mg/L without reserpine. These findings should have implications for understanding the mode of action of PZA and for PZA susceptibility testing.


Type I interferons produced by dendritic cells promote their phenotypic and functional activation.
Resting dendritic cells (DCs) are resident in most tissues and can be activated by environmental stimuli to mature into potent antigen-presenting cells. One important stimulus for DC activation is infection; DCs can be triggered through receptors that recognize microbial components directly or by contact with infection-induced cytokines. We show here that murine DCs undergo phenotypic maturation upon exposure to type I interferons (type I IFNs) in vivo or in vitro. Moreover, DCs either derived from bone marrow cells in vitro or isolated from the spleens of normal animals express IFN-alpha and IFN-beta, suggesting that type I IFNs can act in an autocrine manner to activate DCs. Consistent with this idea, the ability to respond to type I IFN was required for the generation of fully activated DCs from bone marrow precursors, as DCs derived from the bone marrow of mice lacking a functional receptor for type I IFN had reduced expression of costimulatory and adhesion molecules and a diminished ability to stimulate naive T-cell proliferation compared with DCs derived from control bone marrow. Furthermore, the addition of neutralizing anti-IFN-alpha/beta antibody to purified splenic DCs in vitro partially blocked the "spontaneous" activation of these cells, inhibiting the up-regulation of costimulatory molecules, secretion of IFN-gamma, and T-cell stimulatory activity. These results show that DCs both secrete and respond to type I IFN, identifying type I interferons as autocrine DC activators.