Object-based image analysis using multiscale connectivity.
This paper introduces a novel approach for image analysis based on the notion of multiscale connectivity. We use the proposed approach to design several novel tools for object-based image representation and analysis which exploit the connectivity structure of images in a multiscale fashion. More specifically, we propose a nonlinear pyramidal image representation scheme, which decomposes an image at different scales by means of multiscale grain filters. These filters gradually remove connected components from an image that fail to satisfy a given criterion. We also use the concept of multiscale connectivity to design a hierarchical data partitioning tool. We employ this tool to construct another image representation scheme, based on the concept of component trees, which organizes partitions of an image in a hierarchical multiscale fashion. In addition, we propose a geometrically-oriented hierarchical clustering algorithm which generalizes the classical single-linkage algorithm. Finally, we propose two object-based multiscale image summaries, reminiscent of the well-known (morphological) pattern spectrum, which can be useful in image analysis and image understanding applications.


Carvacrol reduces Clostridium difficile sporulation and spore outgrowth in vitro.
Clostridium difficile is an anaerobic spore-forming pathogen that causes a serious toxin-mediated enteric disease in humans. Therapeutic agents that are capable of reducing C. difficile spore production could significantly minimize the transmission and relapse of C. difficile infections. This study investigated the efficacy of a food-grade, plant-derived compound, carvacrol (CR), in reducing C. difficile spore production, germination and spore outgrowth. Two hyper-virulent C. difficile isolates (ATCC BAA 1870 and 1805) were grown with or without a sub-inhibitory concentration (SIC) of CR. Total viable counts and heat-resistant spore counts were determined at different time intervals. Moreover, spores and vegetative cells were visualized using phase-contrast microscopy. To determine the effect of CR on C. difficile germination and spore outgrowth, C. difficile spores were seeded in germination medium with or without the SIC and MIC of CR, and spore germination and spore outgrowth were measured by recording optical density at 600 nm. The effect of CR on C. difficile sporulation genes was also investigated using real-time qPCR. Carvacrol significantly reduced sporulation in C. difficile and down-regulated critical genes involved in spore production (P<0.05). The SIC or MIC of CR did not inhibit C. difficile spore germination; however, the MIC of CR completely inhibited spore outgrowth. The results suggest that CR could potentially be used to control C. difficile by reducing spore production and outgrowth.


Determination of the structure of two novel echistatin variants and comparison of the ability of echistatin variants to inhibit aggregation of platelets from different species.
Two new variants of short disintegrins were purified from the venom of Echis carinatus leakeyi and named echistatin beta and gamma. These proteins were found to be about 85% similar in amino acid sequence to echistatin alpha which has been well studied. The disulphide pattern of echistatin gamma appeared to be identical with that of echistatin alpha. They all contain the adhesive recognition sequence Arg-Gly-Asp (RGD) but inhibit the aggregation of platelets from human and other mammals with different potencies. Echistatin beta and alpha are far more effective on platelets from humans and guinea pigs than those from rabbits and rats whereas echistatin gamma is less discriminating of the platelets of the species tested. This species-dependent platelet sensitivity to echistatin beta and gamma could be attributed to the variations in residues 15, 21, 22 and 27, which are close to or within the RGD loop, rather than to the C-terminal variations after residue 46. Taking advantage of the presence of methionine residues flanking both sides of the ARGDDM motif in echistatin gamma, we deleted this hexapeptide by CNBr cleavage to produce des-(23-28)-echistatin gamma. The modified protein showed c.d. and fluorescent spectra grossly similar to the intact echistatin but its antiplatelet potency decreased more than 200-fold. We thus propose that a favourable conformation of the RGD region is responsible mainly for the high-affinity binding of echistatin to the platelet glycoprotein IIb-IIIa as shown previously for the binding of medium-size disintegrin.


A probabilistic sampling method (PSM) for estimating geographic distance to health services when only the region of residence is known.
The need to estimate the distance from an individual to a service provider is common in public health research. However, estimated distances are often imprecise and, we suspect, biased due to a lack of specific residential location data. In many cases, to protect subject confidentiality, data sets contain only a ZIP Code or a county. This paper describes an algorithm, known as "the probabilistic sampling method" (PSM), which was used to create a distribution of estimated distances to a health facility for a person whose region of residence was known, but for which demographic details and centroids were known for smaller areas within the region. From this distribution, the median distance is the most likely distance to the facility. The algorithm, using Monte Carlo sampling methods, drew a probabilistic sample of all the smaller areas (Census blocks) within each participant's reported region (ZIP Code), weighting these areas by the number of residents in the same age group as the participant. To test the PSM, we used data from a large cross-sectional study that screened women at a clinic for intimate partner violence (IPV). We had data on each woman's age and ZIP Code, but no precise residential address. We used the PSM to select a sample of census blocks, then calculated network distances from each census block's centroid to the closest IPV facility, resulting in a distribution of distances from these locations to the geocoded locations of known IPV services. We selected the median distance as the most likely distance traveled and computed confidence intervals that describe the shortest and longest distance within which any given percent of the distance estimates lie. We compared our results to those obtained using two other geocoding approaches. We show that one method overestimated the most likely distance and the other underestimated it. Neither of the alternative methods produced confidence intervals for the distance estimates. The algorithm was implemented in R code. The PSM has a number of benefits over traditional geocoding approaches. This methodology improves the precision of estimates of geographic access to services when complete residential address information is unavailable and, by computing the expected distribution of possible distances for any respondent and associated distance confidence limits, sensitivity analyses on distance access measures are possible. Faulty or imprecise distance measures may compromise decisions about service location and misdirect scarce resources.


Parenting stress trajectories in mothers of very preterm infants to 2 years.
To examine levels of parenting stress in mothers of preterm and term infants when the children were 2 years old; to determine the trajectory of stress over three time periods and to examine the association of maternal and neonatal factors and developmental outcomes with parenting stress. It is a prospective longitudinal study to determine parenting stress in mothers of preterm and term infants with outcomes having been previously obtained at 4 and 12 months. At 2 years, 79 preterm mothers (96 babies) and 64 term mothers (77 babies) participated. The mothers completed the Parenting Stress Index-Short Form (PSI-SF), the Depression, Anxiety, Stress Scale (DASS) and the Child Behaviour Checklist (CBCL). The infants had a neurological examination and the Bayley-III scales were administered. The mean total PSI-SF at 2 years was significantly higher for the preterm group compared with the term group of mothers (p=0.007). There was a significant increase in the mean total PSI over time for the preterm mothers (p<0.001). For mothers at 2 years, there was an association with high levels of parenting stress and abnormal scores on the DASS (p<0.001) and high total T-scores on the CBCL (internalising p<0.001; externalising p=0.006). There was no association between parenting stress and maternal demographics, neonatal factors or Bayley-III results. Parenting stress in mothers of preterm infants continues to be high at 2 years having increased over time. Maternal mental health problems and infant behavioural issues contribute to the stress.


Squash smear cytology of Langerhans cell histiocytosis.
Squash smear cytology of Langerhans cell histiocytosis (LCH) has rarely been reported. We described squash cytological findings of cranial LCH. Additionally, based on recent data that suggests an association of LCH with either viral infection or genetic alteration, we investigated the presence of several viruses or mutation of TP53 and BRAF in LCH tissue samples. Intraoperative squash smears of a small tissue fragment excised from the lesion demonstrated a mixed population of eosinophils, neutrophils, small lymphocytes and a high content of histiocytes. The histiocytes possessed abundant dense cytoplasm with round cell shape and eccentrically located nuclei with fine chromatin, delicate nuclear membranes and prominent nuclear grooves, indentations and pseudoinclusions. The cytologic features were consistent with Langerhans cells (LCs). Subsequent histopathologic examination confirmed the diagnosis of LCH. Immunohistochemically, the LCs were positive for S-100, CD1a and langerin, but negative for adenovirus, CMV, EBV, HHV-8, HPV, HSV, SV 40 and p53. BRAF V600E mutation was absent. Our findings did not support the role of viruses and genetic abnormalities in the pathogenesis of LCH. In summary, the presence of a mixed population of inflammatory cells and a high content of histiocytes with characteristic cytomorphology, along with radiologic evidence and appropriate clinical findings, is highly suggestive of LCH on the intraoperative squash smears. Awareness of characteristic cytological features of LCH is necessary for rapid and accurate diagnosis. Squash smear cytology is a potentially useful tool in the intraoperative diagnosis of LCH.


Interventions for nausea and vomiting in early pregnancy.
Nausea, retching and vomiting are very commonly experienced by women in early pregnancy. There are considerable physical and psychological effects on women who experience these symptoms. This is an update of a review of interventions for nausea and vomiting in early pregnancy previously published in 2003. To assess the effectiveness and safety of all interventions for nausea, vomiting and retching in early pregnancy, up to 20 weeks' gestation. We searched the Cochrane Pregnancy and Childbirth Group's Trials Register (28 May 2010). All randomised controlled trials of any intervention for nausea, vomiting and retching in early pregnancy. We excluded trials of interventions for hyperemesis gravidarum which are covered by another review. We also excluded quasi-randomised trials and trials using a crossover design. Four review authors, in pairs, reviewed the eligibility of trials and independently evaluated the risk of bias and extracted the data for included trials. Twenty-seven trials, with 4041 women, met the inclusion criteria. These trials covered many interventions, including acupressure, acustimulation, acupuncture, ginger, vitamin B6 and several antiemetic drugs. We identified no studies of dietary or other lifestyle interventions. Evidence regarding the effectiveness of P6 acupressure, auricular (ear) acupressure and acustimulation of the P6 point was limited. Acupuncture (P6 or traditional) showed no significant benefit to women in pregnancy. The use of ginger products may be helpful to women, but the evidence of effectiveness was limited and not consistent. There was only limited evidence from trials to support the use of pharmacological agents including vitamin B6, and anti-emetic drugs to relieve mild or moderate nausea and vomiting. There was little information on maternal and fetal adverse outcomes and on psychological, social or economic outcomes. We were unable to pool findings from studies for most outcomes due to heterogeneity in study participants, interventions, comparison groups, and outcomes measured or reported. The methodological quality of the included studies was mixed. Given the high prevalence of nausea and vomiting in early pregnancy, health professionals need to provide clear guidance to women, based on systematically reviewed evidence. There is a lack of high-quality evidence to support that advice. The difficulties in interpreting the results of the studies included in this review highlight the need for specific, consistent and clearly justified outcomes and approaches to measurement in research studies.


Altered temporal characteristics of the rectoanal inhibitory reflex in patients with abdominal distension.
The rectoanal inhibitory reflex (RAIR) is important in gas and stool evacuation. We examined RAIR features in patients with chronic constipation who exhibited bloating with and without abdominal distension, to determine whether alterations in RAIR may be a factor in the pathogenesis of abdominal distension. Seventy-five female patients with chronic constipation with or without abdominal distension were included in the study. The presence or absence of abdominal distension was assessed according to the Rome II questionnaire. All patients underwent both RAIR and rectal sensitivity testing, and specific RAIR parameters were analyzed. Patients were divided into two groups: abdominal bloating with distension (D, n = 55) and abdominal bloating without distension (ND, n = 20). D had a longer time to the onset of anal sphincter inhibition (latency of inhibition) (P = 0.03) compared with ND. In logistic regression analysis, a combination of age, latency of inhibition and the time measured from onset of inhibition to the point of maximum inhibition predicted abdominal distension (P = 0.002). There were no differences between groups for the time from point of maximum inhibition to recovery and for the percentage of internal anal sphincter relaxation. This is the first study to examine the role of RAIR in patients with abdominal distension. Female patients with constipation and abdominal distension exhibited differences in the temporal characteristics of, but not in the degree of, anal sphincter relaxation compared with patients without distension. Since this study was uncontrolled, further studies are necessary to determine the contribution of altered anorectal reflexes to abdominal distension.


Molecular engineering of polymeric supra-amphiphiles.
Polymeric supra-amphiphiles are amphiphiles that are fabricated by linking polymeric segments, or small molecules and polymeric segments, by noncovalent interactions or dynamic covalent bonds. Compared with conventional amphiphilic polymers, polymeric supra-amphiphiles are advantageous in that they possess dynamic features and their preparation may be to some extent more facile. Moreover, polymeric supra-amphiphiles are endowed with richer structure and higher stability compared with small-molecule supra-amphiphiles. Owing to these properties, polymeric supra-amphiphiles have so far shown great promise as surfactants, nanocarriers and in therapies. In this tutorial review, recent work on polymeric supra-amphiphiles, from molecular architectures to functional assemblies, is presented and summarized. Different polymeric supra-amphiphile topologies and related applications are highlighted. By combining polymer chemistry with supramolecular chemistry and colloid science, we anticipate that the study of polymeric supra-amphiphiles will promote the continued development of the molecular engineering of functional supramolecular systems, and lead to practical applications, especially in drug delivery.


Wnt signaling, Ca2+, and cyclic GMP: visualizing Frizzled functions.
Wnts control the specification of cell fate, cell adhesion, migration, polarity, and proliferation. Their roles in development have been probed in fruit flies, nematodes, zebrafish, frogs, and mice. Some Wnts inhibit the degradation of beta-catenin, which can regulate transcription of specific genes. Other Wnts exert their influences in other ways, such as increasing intracellular concentrations of Ca2+ and decreasing intracellular concentrations of cyclic guanosine monophosphate (cGMP). Heterotrimeric guanine nucleotide-binding proteins (G proteins) and RGS proteins have been implicated in Wnt signaling. Wnt regulation of intracellular Ca2+ and cGMP levels requires the G protein transducin and a cGMP-specific phosphodiesterase, which are major elements in signaling of the visual pathway.


Bone morphogenetic protein 3 controls insulin gene expression and is down-regulated in INS-1 cells inducibly expressing a hepatocyte nuclear factor 1A-maturity-onset diabetes of the young mutation.
Inactivating mutations in the transcription factor hepatocyte nuclear factor (HNF) 1A cause HNF1A-maturity-onset diabetes of the young (HNF1A-MODY), the most common monogenic form of diabetes. To examine HNF1A-MODY-induced defects in gene expression, we performed a microarray analysis of the transcriptome of rat INS-1 cells inducibly expressing the common hot spot HNF1A frameshift mutation, Pro291fsinsC-HNF1A. Real-time quantitative PCR (qPCR), Western blotting, immunohistochemistry, reporter assays, and chromatin immunoprecipitation (ChIP) were used to validate alterations in gene expression and to explore biological activities of target genes. Twenty-four hours after induction of the mutant HNF1A protein, we identified a prominent down-regulation of the bone morphogenetic protein 3 gene (Bmp-3) mRNA expression. Reporter assays, qPCR, and Western blot analysis validated these results. In contrast, inducible expression of wild-type HNF1A led to a time-dependent increase in Bmp-3 mRNA and protein levels. Moreover, reduced protein levels of BMP-3 and insulin were detected in islets of transgenic HNF1A-MODY mice. Interestingly, treatment of naïve INS-1 cells or murine organotypic islet cultures with recombinant human BMP-3 potently increased their insulin levels and restored the decrease in SMAD2 phosphorylation and insulin gene expression induced by the HNF1A frameshift mutation. Our study suggests a critical link between HNF1A-MODY-induced alterations in Bmp-3 expression and insulin gene levels in INS-1 cells and indicates that the reduced expression of growth factors involved in tissue differentiation may play an important role in the pathophysiology of HNF1A-MODY.


Cortical Control of Striatal Dopamine Transmission via Striatal Cholinergic Interneurons.
Corticostriatal regulation of striatal dopamine (DA) transmission has long been postulated, but ionotropic glutamate receptors have not been localized directly to DA axons. Striatal cholinergic interneurons (ChIs) are emerging as major players in striatal function, and can govern DA transmission by activating nicotinic receptors (nAChRs) on DA axons. Cortical inputs to ChIs have historically been perceived as sparse, but recent evidence indicates that they strongly activate ChIs. We explored whether activation of M1/M2 corticostriatal inputs can consequently gate DA transmission, via ChIs. We reveal that optogenetic activation of channelrhodopsin-expressing corticostriatal axons can drive striatal DA release detected with fast-scan cyclic voltammetry and requires activation of nAChRs on DA axons and AMPA receptors on ChIs that promote short-latency action potentials. By contrast, DA release driven by optogenetic activation of intralaminar thalamostriatal inputs involves additional activation of NMDA receptors on ChIs and action potential generation over longer timescales. Therefore, cortical and thalamic glutamate inputs can modulate DA transmission by regulating ChIs as gatekeepers, through ionotropic glutamate receptors. The different use of AMPA and NMDA receptors by cortical versus thalamic inputs might lead to distinct input integration strategies by ChIs and distinct modulation of the function of DA and striatum.


Factors affecting exclusive breastfeeding of healthy babies aged zero to four months: a community-based study of Turkish women.
The purpose of this study was to determine the factors that affect exclusive breastfeeding of healthy babies aged 0-4 months. Both the World Health Organization (WHO) and UNICEF recommend feeding babies of 0-6 months exclusively with breast milk and starting complementary foods after the sixth month. In Turkey, however, a vast majority of babies 1-5 months of age (89.4%) are given complementary foods. This cross-sectional study was conducted in central Ankara province, with a sampling of 514 individuals who were selected using the convenience sampling method. Of the 514 mothers who participated in my research, 260 (50.6%) were found to be feeding their babies exclusively with breast milk; 77 (15.0%), with breast milk + water; 87 (16.9%), with breast milk + baby formula; 70 (13.6%), with breast milk + baby formula + other foods; and 20 (3.9%), baby formula + other foods. Based on multivariate logistic regression analysis results, the mother's employment [odds ratio (OR) = 0.488; 95% confidence interval (CI) = 0.288-0.827) considerably reduced the incidence of complementary foods, while frequent crying of the baby (OR = 1.687; 95% CI = 1.125-2.530) significantly increased the use of supplementary foods in infant nutrition. This study concluded that frequent crying of the baby increases the likelihood of giving the baby complementary foods. Midwives and nurses can encourage exclusive breastfeeding behaviour by providing individual education and counselling to women whose babies cry frequently. Exclusive breastfeeding of babies aged 0-6 months is crucial for the development and growth of the baby and instrumental in reducing infant morbidities and mortalities. One factor that increases the likelihood of provision of complementary foods is frequent crying of the baby. Midwives and nurses can encourage exclusive breastfeeding behaviour by providing individual education and counselling to women whose babies cry frequently.


Dry needling of the trapezius muscle in office workers with neck pain: a randomized clinical trial.
Neck pain is a frequent complaint in office workers. This pain can be caused by myofascial trigger points (MTrPs) in the trapezius muscle. This study aimed to determine the effectiveness of deep dry needling (DDN) of active MTrPs in the trapezius muscle. A randomized, single blinded clinical trial was carried out at the Physical Therapy Department at Physiotherapy in Women's Health Research Group at Physical Therapy Department of University of Alcalá, in Alcalá de Henares, Madrid, Spain. Forty-four office workers with neck pain and active MTrPs in the trapezius muscle were randomly allocated to either the DDN or the control group (CG). The participants in the DDN group were treated with DDN of all MTrPs found in the trapezius muscle. They also received passive stretch of the trapezius muscle. The CG received the same passive stretch of the trapezius muscle only. The primary outcome measure was subjective pain intensity, measured using a visual analogue scale (VAS). Secondary outcomes were pressure pain threshold (PPT), cervical range of motion (CROM) and muscle strength. Data were collected at baseline, after interventions and 15 days after the last treatment. Differences were found between the DDN group and the CG for the VAS (P < 0.001), PPT (P < 0.001), range of motion (AROM) (P < 0.05) and strength (P < 0.05) after intervention and at the 15-day follow-up. Deep dry needling and passive stretch seems to be more effective than passive stretch only. The effects are maintained in the short term. The results support the use of DDN in the management of trapezius muscle myofascial pain syndrome in neck pain.


Scurvy: a disease almost forgotten.
Although much decreased in prevalence, scurvy still exists in industrialized societies. Few recent large studies have examined its pathogenesis, signs, and symptoms. After we diagnosed scurvy in a 77-year-old female patient in 2003, we conducted a retrospective records review to identify patients with scurvy treated between 1976 and 2002 at Mayo Clinic (Scottsdale, Arizona; Rochester, Minnesota; or Jacksonville, Florida). We also searched the English-language medical literature for published reports on scurvy. In addition to our patient, seven of 11 patients whose records in the institutional database mentioned vitamin C deficiency were women. The age ranged from a neonate to 77 years (mean, 48 years). The most common associated causes were concomitant gastrointestinal disease, poor dentition, food faddism, and alcoholism. Vitamin or mineral deficiencies other than vitamin C deficiency were also found in our patients who had scurvy. The most common symptoms were bruising, arthralgias, or joint swelling. The most common signs were pedal edema, bruising, or mucosal changes. Four patients had vague symptoms of myalgias and fatigue without classic findings, and five had concomitant nutritional deficiencies. Follow-up available for six of 12 patients treated by vitamin C supplementation showed complete resolution of symptoms in five. Patients with scurvy may present with classic symptoms and signs or with nonspecific clinical symptoms and an absence of diagnostically suggestive physical findings. Concomitant deficiency states occur not uncommonly. Taking a thorough dietary history and measuring serum ascorbic acid levels should be considered for patients with classic signs and symptoms, nonspecific musculoskeletal complaints, or other vitamin or mineral deficiencies.


Coupled fenton-denitrification process for the removal of organic matter and total nitrogen from coke plant wastewater.
This work assesses the feasibility of applying a Coupled Fenton-Denitrification (CFD) process for the treatment of wastewater from a coking plant. This highly toxic effluent is characterized by comparable carbon and nitrogen contents and it is usually released into the treatment system at well above room temperature. Recalcitrant organic matter can be easily removed in a first step using Fenton treatment. Working at 50 °C, pH0: 3, and a wastewater obtained from a coking plant, the stoichiometric amount of H2O2 relative to COD and a H2O2/Fe2+ weight ratio of 50, around 60% of carbon load was mineralized whereas H2O2 was completely depleted. However, no changes were observed in the total nitrogen content. A subsequent denitrification stage led to an additional 80% TOC (overall above 90%) and 75% Total Nitrogen removal. This was done in a batch bioreactor at room temperature over 72 h, using a 40-day pre-acclimated denitrifying biomass. These results point to the possibility of designing a combined chemical oxidation and biological treatment to deal with complex effluents containing refractory organic matter including high concentrations of nitrogen species.


"To observe well ... and thence to make himself rules": John Locke's principles and practice of child healthcare.
It is often forgotten that the philosopher John Locke (1632-1704) was a highly regarded physician with a lifelong interest in medicine and was frequently consulted on medical matters, including the health of children. This child health aspect in Locke's history has been largely ignored, with even modern commentaries on Locke and medicine giving it only a cursory mention. However, it is clear that, in child health, Locke's influence is far more substantial than GF Still's and George Jackson's opinions, which limited Locke solely to Thoughts concerning education (1692/3). That a fundamental reappraisal of Locke's role in child healthcare is necessary and that his place as a pioneer of modern child healthcare needs to be proclaimed are emphasised here. As modern day child healthcare has evolved to embrace advocacy and learning disability, Locke's importance through his influence on paediatrics, child healthcare and human rights becomes more evident. Locke's influence in child healthcare comes not only through his other celebrated philosophical writings, but also through extensive personal correspondence and case records. As well as throwing light onto the 17th century aspects of child healthcare, Locke, through his enquiry and self-evident humility in his correspondence on medical matters, inspires and educates us with his pragmatic approach to the practice of medicine.


The Arabidopsis ATK1 gene is required for spindle morphogenesis in male meiosis.
The spindle plays a central role in chromosome segregation during mitosis and meiosis. In particular, various kinesins are thought to play crucial roles in spindle structure and function in both mitosis and meiosis of fungi and animals. A group of putative kinesins has been previously identified in Arabidopsis, called ATK1-ATK4 (previously known as KATA-KATD), but their in vivo functions have not been tested with genetic studies. We report here the isolation and characterization of a mutant, atk1-1, which has a defective ATK1 gene. The atk1-1 mutant was identified in a collection of Ds transposon insertion lines by its reduced fertility. Reciprocal crosses between the atk1-1 mutant and wild type showed that only male fertility was reduced, not female fertility. Molecular analyses, including revertant studies, indicated that the Ds insertion in the ATK1 gene was responsible for the fertility defect. Light microscopy revealed that, in the atk1-1 mutant, male meiosis was defective, producing an abnormal number of microspores of variable sizes. Further cytological studies indicated that meiotic chromosome segregation and spindle organization were both abnormal in the mutant. Specifically, the atk1-1 mutant male meiotic cells had spindles that were broad, unfocused and multi-axial at the poles at metaphase I, unlike the typical fusiform bipolar spindle found in the wild-type metaphase I cells. Therefore, the ATK1 gene plays a crucial role in spindle morphogenesis in male Arabidopsis meiosis.


Multiple protein-binding sites in an intracisternal A particle long terminal repeat.
The long terminal repeats (LTRs) of cloned intracisternal A particles (IAPs) can function as effective promoters in heterologous and homologous cell types (K. K. Lueders, J. W. Fewell, E. L. Kuff, and T. Koch, Mol. Cell. Biol. 4:2128-2135, 1984) and respond to transcriptional factors induced by various nuclear oncogene products (S. Luria and M. Horowitz, J. Virol. 57:998-1003, 1986). Using the first 139 base pairs of the U3 region of a cloned mouse IAP LTR as probe, we demonstrated multiple exonuclease III stop sites which appeared specifically in the presence of nuclear extract protein. Various extracts gave similar footprints, but the amount of nuclear protein required varied up to 10-fold. Cell lines transformed with known nuclear oncogenes, such as adenovirus E1a and E1b (293 cells), simian virus 40 large T antigen (COS7 cells), and c-myc (MOPC-315 cells) had more and/or higher-affinity factors for the IAP LTR than extracts from HeLa, CV1, and NIH 3T3 cells did. DNase I footprinting revealed at least five distinct protein-binding domains within the 139-base-pair region. These domains correspond to segments of highly conserved nucleotide sequence among a number of IAP LTRs. Gel retardation studies with oligonucleotides encompassing the DNase I footprint sites showed that the nuclear factors are present in different proportions and different absolute levels in extracts from different cell types. Moreover, the oligonucleotide probes indicate that individual motifs can be occupied independently of one another. Three of the DNase I footprints include a sequence with homology to the simian virus 40 core enhancer and sequence motifs that closely resemble the binding sites for transcription factors SP1 and AP-1. The other two binding sites are not obviously related to previously recognized motifs. The multiple protein-binding sites in close proximity indicate the complex regulatory mechanism for IAP transcription.


Costs of irritable bowel syndrome in the UK and US.
Irritable bowel syndrome (IBS) is one of the most common functional gastrointestinal disorders, with an estimated prevalence rate in the general population of 10-15% in industrialised countries. Although IBS is not a life-threatening disease, it contributes significantly to a large segment of healthcare resource consumption. This review provides an overview of studies addressing the direct and indirect costs of IBS in the US and the UK. A systematic literature search was conducted in MEDLINE and the Cochrane library; additionally, all reference lists covering the years from 1960 to May 2004 were scanned. Twenty-four publications for the US and the UK, published between 1991 and 2003, were identified: 6 were excluded, 18 were included. Data for the UK, US and UK + US were reported in 5, 11 and 2 publications, respectively. Total direct cost estimates per patient per year ranged from US 348 dollars to US 8750 dollars (calculated for year 2002). The average number of days off work per year because of IBS was between 8.5 and 21.6; indirect costs ranged from US 355 dollars to US 3344 dollars. The total costs and cost components of IBS are influenced by several factors: features of the investigated patient group (age, limitation to healthcare seekers or all IBS patients, comorbidity, severity of symptoms), database used, method of data collection (retrospective or prospective, varying cost components, time-point of data collection in relation to index-date of IBS diagnosis, method of cost calculation [incidence or prevalence based]) and different healthcare systems in the US and the UK. These factors led to the incomparability of published data, thus no comprehensive picture can be drawn of the total costs related to IBS in the UK and US. Data underline the magnitude of the economic impact of IBS in the UK and US, which is increased by a factor of 1.1-6.0, compared with matched non-IBS control groups. IBS contributes both direct and indirect costs to the total healthcare bill. Further studies should take influencial factors into account and report related data carefully in order to provide useful and comparable published cost data. Additionally, further research on the cost effectiveness of diagnostic procedures and therapies in IBS is required to define strategies to help IBS patients improve their quality of life and reduce related costs.


Holistic services for people with advanced disease and chronic breathlessness: a systematic review and meta-analysis.
Breathlessness is a common, distressing symptom in people with advanced disease and a marker of deterioration. Holistic services that draw on integrated palliative care have been developed for this group. This systematic review aimed to examine the outcomes, experiences and therapeutic components of these services. Systematic review searching nine databases to June 2017 for experimental, qualitative and observational studies. Eligibility and quality were independently assessed by two authors. Data on service models, health and cost outcomes were synthesised, using meta-analyses as indicated. Data on recipient experiences were synthesised thematically and integrated at the level of interpretation and reporting. From 3239 records identified, 37 articles were included representing 18 different services. Most services enrolled people with thoracic cancer, involved palliative care staff and comprised 4-6 contacts over 4-6 weeks. Commonly used interventions included breathing techniques, psychological support and relaxation techniques. Meta-analyses demonstrated reductions in Numeric Rating Scale distress due to breathlessness (n=324; mean difference (MD) -2.30, 95% CI -4.43 to -0.16, p=0.03) and Hospital Anxiety and Depression Scale (HADS) depression scores (n=408, MD -1.67, 95% CI -2.52 to -0.81, p<0.001) favouring the intervention. Statistically non-significant effects were observed for Chronic Respiratory Questionnaire (CRQ) mastery (n=259, MD 0.23, 95% CI -0.10 to 0.55, p=0.17) and HADS anxiety scores (n=552, MD -1.59, 95% CI -3.22 to 0.05, p=0.06). Patients and carers valued tailored education, self-management interventions and expert staff providing person-centred, dignified care. However, there was no observable effect on health status or quality of life, and mixed evidence around physical function. Holistic services for chronic breathlessness can reduce distress in patients with advanced disease and may improve psychological outcomes of anxiety and depression. Therapeutic components of these services should be shared and integrated into clinical practice. CRD42017057508.


Identification of a potent and selective pharmacophore for Cdc25 dual specificity phosphatase inhibitors.
Small molecules provide powerful tools to interrogate biological pathways but many important pathway participants remain refractory to inhibitors. For example, Cdc25 dual-specificity phosphatases regulate mammalian cell cycle progression and are implicated in oncogenesis, but potent and selective inhibitors are lacking for this enzyme class. Thus, we evaluated 10,070 compounds in a publicly available chemical repository of the National Cancer Institute for in vitro inhibitory activity against oncogenic, full-length, recombinant human Cdc25B. Twenty-one compounds had mean inhibitory concentrations of <1 microM; >75% were quinones and >40% were of the para-naphthoquinone structural type. Most notable was NSC 95397 (2,3-bis-[2-hydroxyethylsulfanyl]-[1,4]naphthoquinone), which displayed mixed inhibition kinetics with in vitro K(i) values for Cdc25A, -B, and -C of 32, 96, and 40 nM, respectively. NSC 95397 was more potent than any inhibitor of dual specificity phosphatases described previously and 125- to 180-fold more selective for Cdc25A than VH1-related dual-specificity phosphatase or protein tyrosine phosphatase 1b, respectively. Modification of the bis-thioethanol moiety markedly decreased enzyme inhibitory activity, indicating its importance for bioactivity. NSC 95397 showed significant growth inhibition against human and murine carcinoma cells and blocked G(2)/M phase transition. A potential Cdc25 site of interaction was postulated based on molecular modeling with these quinones. We propose that inhibitors based on this chemical structure could serve as useful tools to probe the biological function of Cdc25.


Characterization of the sdw1 semi-dwarf gene in barley.
The dwarfing gene sdw1 has been widely used throughout the world to develop commercial barley varieties. There are at least four different alleles at the sdw1 locus. Mutations in the gibberellin 20-oxidase gene (HvGA20ox2) resulted in multiple alleles at the sdw1 locus. The sdw1.d allele from Diamant is due to a 7-bp deletion in exon 1, while the sdw1.c allele from Abed Denso has 1-bp deletion and a 4-bp insertion in the 5' untranslated region. The sdw1.a allele from Jotun resulted from a total deletion of the HvGA20ox2 gene. The structural changes result in lower gene expression in sdw1.d and lack of expression in sdw1.a. There are three HvGA20ox genes in the barley genome. The partial or total loss of function of the HvGA20ox2 gene could be compensated by enhanced expression of its homolog HvGA20ox1and HvGA20ox3. A diagnostic molecular marker was developed to differentiate between the wild-type, sdw1.d and sdw1.a alleles and another molecular marker for differentiation of sdw1.c and sdw1.a. The markers were further tested in 197 barley varieties, out of which 28 had the sdw1.d allele and two varieties the sdw1.a allele. To date, the sdw1.d and sdw1.a alleles have only been detected in the modern barley varieties and lines. The results provided further proof that the gibberellin 20-oxidase gene (HvGA20ox2) is the functional gene of the barley sdw1 mutants. Different deletions resulted in different functional alleles for different breeding purposes. Truncated protein could maintain partial function. Partial or total loss of function of the HvGA20ox2 gene could be compensated by enhanced expression of its homolog HvGA20ox1 and HvGA20ox3.


Surveillance for Hepatocellular Carcinoma Reduces Mortality: an Inverse Probability of Treatment Weighted Analysis.
Evidence supporting benefit of hepatocellular carcinoma (HCC) surveillance in reducing mortality is not well-established. The effect of HCC surveillance in reducing mortality was assessed by an inverse probability of treatment weighting (IPTW)-based analysis controlled for inherent bias and confounders in observational studies. This retrospective cohort study was conducted on 446 patients diagnosed with HCC between 2007 and 2013 at a major referral center. Surveillance was defined as having at least 1 ultrasound test within a year before HCC diagnosis. Primary outcome was survival estimated using the Kaplan-Meier method with lead-time bias adjustment and compared using the log-rank test. Hazard ratio (HR) and 95% confidence interval (CI) were computed using conventional Cox and weighted Cox proportional hazards analysis with IPTW adjustment. Of the 446 patients, 103 (23.1%) were diagnosed with HCC through surveillance. The surveillance group had more patients with the Barcelona-Clinic Liver Cancer stage A (80.6% vs. 33.8%, P < 0.0001), more patients eligible for potentially curative treatment (73.8% vs. 44.9%, P < 0.0001), and longer median survival (49.6 vs. 15.9 months, P < 0.0001). By conventional multivariate Cox analysis, HR (95% CI) of surveillance was 0.63 (0.45-0.87), P = 0.005. The estimated effect of surveillance remained similar in the IPTW-adjusted Cox analysis (HR: 0.57; 95% CI: 0.43-0.76, P < 0.001). HCC surveillance by ultrasound is associated with a 37% reduction in mortality. Even though surveillance is recommended in all guidelines, but in practice, it is underutilized. Interventions are needed to increase surveillance rate for improving HCC outcome.


Sex-specific phenotypical and functional differences in peripheral human Vgamma9/Vdelta2 T cells.
Vgamma9/Vdelta2 T cells constitute a minor proportion of human peripheral blood T cells that can expand rapidly upon infection with microbial pathogens. Vgamma9/Vdelta2 T cell numbers change characteristically with age, rising from birth to puberty and gradually decreasing again beyond 30 years of age. In adults, female blood donors have significantly higher levels than males, implying that circulating Vgamma9/Vdelta2 T cells in women remain elevated for a longer period in life and drop less strikingly than in men. This loss in men is accompanied by a substantial depletion of CD27- CD45RA- and CD27- CD45RA+ effector T cells and a parallel increase in CD27+ CD45RA- central memory T cells while in women, the distribution of Vgamma9/Vdelta2 T cell subsets remains virtually unchanged. The phenotypical conversion in men older than 30 years is mirrored by an increased proliferative response of Vgamma9/Vdelta2 T cells and a reduced interferon-gamma secretion upon stimulation with isopentenyl pyrophosphate in vitro.


Dietary n-3 HUFA affects mitochondrial fatty acid beta-oxidation capacity and susceptibility to oxidative stress in Atlantic salmon.
Atlantic salmon (Salmo salar) (90 g) were fed four different diets for 21 weeks (final weight 344 g). The levels of n-3 highly unsaturated fatty acids (HUFA) ranged from 11% of the total fatty acids (FA) in the low n-3 diet to 21% in the intermediate n-3 diet, to 55 and 58% in the high n-3 diets. The high n-3 diets were enriched with either docosahexaenoic acid (DHA) or eicosapentaenoic acid (EPA). Increasing dietary levels of n-3 HUFA led to increasing percentages (from 31 to 52%) of these FA in liver lipids. The group fed the highest level of DHA had higher expressions of peroxisome proliferator-activated receptor (PPAR) beta and the FA beta-oxidation genes acyl-CoA oxidase (ACO) and carnitine palmitoyltransferase (CPT)-II, compared to the low n-3 groups. The high n-3 groups had reduced activity of mitochondrial cytochrome c oxidase and beta-oxidation capacity, together with increased activities of superoxide dismutase (SOD) and caspase-3 activities. In the group fed the highest level of n-3 HUFA, decreased percentages of major phospholipids (PL) in the mitochondrial and microsomal membranes of the liver were also apparent. The percentage of mitochondrial cardiolipin (Ptd(2)Gro) was 3.1 in the highest n-3 group compared to 6.6 in the intermediate group. These data clearly show an increased incidence of oxidative stress in the liver of fish fed the high n-3 diets.


Disease burden of adult lung cancer and ischaemic heart disease from passive tobacco smoking in China.
To address the health hazards tobacco smoking imposes upon non-smokers in China, this paper estimates the burden of diseases in adults from passive tobacco smoking for two major diseases--lung cancer and ischaemic heart disease (IHD). The disease burden was estimated in terms of both premature mortality and disability adjusted life years (DALYs), a measure that accounts for both the age at death and the severity of the morbidity. Passive smoking caused more than 22,000 lung cancer deaths in 2002 according to these estimates. When the toll of disability is added to that of mortality, passive smoking was responsible for the loss of nearly 230,000 years of healthy life from lung cancer. Using the evidence from other countries that links IHD to passive smoking, we estimated that approximately 33,800 IHD deaths could be attributable to passive smoking in China in 2002. Passive smoking is also responsible for the loss of more than one quarter of a million years of healthy life from IHD. Although most of the disease burden caused by active smoking occurs among men, women bear nearly 80% of the total burden from passive smoking. The number of deaths among women caused by passive smoking is about two-thirds of that caused by smoking for the two diseases we examined. Even without considering the passive smoking risks for other diseases and among children that have been documented in other countries, passive smoking poses serious health hazards for non-smokers, especially for adult female non-smokers in China, adding more urgency to the need for measures to be taken immediately to protect the health of non-smokers and curb the nation's tobacco epidemic.


Human Follicle-Stimulating Hormone ß Subunit Expression Depends on FOXL2 and SMAD4.
Follicle-stimulating hormone (FSH), an essential regulator of mammalian fertility, is synthesized by pituitary gonadotrope cells in response to activins. In mice, activins signal via SMAD3, SMAD4, and FOXL2 to regulate transcription of the FSHβ subunit (Fshb) gene. Gonadotrope-specific deletion of Foxl2, alone or in combination with Smad4, renders mice FSH-deficient. Whether human FSHB expression is similarly regulated is not known. Here, we used a combination of transgenic and conditional knockout mouse strains to assess the roles of activins, FOXL2, and SMAD4 in regulation of the human FSHB gene. First, we cultured pituitaries from mice harboring a human FSHB transgene (hFSHB mice) and measured both murine Fshb and human FSHB messenger ribonucleic acid (mRNA) expression in response to exogenous activins or two antagonists of endogenous activin-like signaling (follistatin-288 and SB431542). Both murine Fshb and human FSHB expression were stimulated by activins and reduced by the inhibitors. Next, we analyzed human FSHB expression in hFSHB mice carrying floxed Foxl2 and Smad4 alleles. Cre-mediated ablation of FOXL2 and SMAD4 strongly reduced basal and activin-stimulated murine Fshb and human FSHB expression in cultured pituitaries. Finally, the hFSHB transgene was previously shown to rescue FSH production and fertility in Fshb knockout mice. However, gonadotrope-specific Foxl2/Smad4 knockout females carrying the hFSHB transgene have significantly reduced murine Fshb and human FSHB pituitary mRNA levels and are hypogonadal. Collectively, these data suggest that similar to Fshb regulation in mice, FOXL2 and SMAD4 play essential roles in human FSHB expression.


Expert panel vs decision-analysis recommendations for postdischarge coronary angiography after myocardial infarction.
Expert panels and decision-analytic techniques are increasingly used to determine the appropriateness of medical interventions, but these 2 approaches use different methods to process evidence. To compare expert panel appropriateness ratings of coronary angiography after myocardial infarction (from the time of hospital discharge to 12 weeks after infarction) with the health gains and cost-effectiveness predicted by a decision-analytic model. Comparison of the degree of importance of the clinical variables considered in expert panel appropriateness ratings vs a previously published decision-analytic model. Identification of 36 clinical scenarios from the expert panel that could be simulated by the decision-analytic model. Appropriateness score and appropriateness classification (expert panel) vs gain in quality-adjusted life-years (QALYs) and incremental cost-effectiveness ratio (decision-analytic model). The most important clinical variables were similar in the 2 approaches, with the exercise tolerance test result exerting the greatest leverage on strength of recommendation for angiography. Among the expert panel clinical scenarios considered to be appropriate for coronary angiography that could be simulated in the decision-analysis model, the median (interquartile range) health gain and incremental cost-effectiveness ratio were 0.59 (0.41-0.76) QALYs and $27000 ($23000-$35000) per QALY gained, respectively. Among the clinical scenarios that expert panels considered inappropriate, the corresponding medians (interquartile ranges) were 0.24 (0.19-0.34) QALYs and $54000 ($36000-$58000) per QALY gained. The Spearman rank correlation between appropriateness score and QALY gain was 0.58 (P<.001) and between appropriateness score and estimated incremental cost-effectiveness ratios was -0.66 (P<.001). For the 36 expert panel scenarios that could be simulated by the decision-analytic model, there was moderate to good agreement between the appropriateness score and both the health gain and the incremental cost-effectiveness ratio of coronary angiography compared with no angiography in the convalescent phase of acute myocardial infarction, but several scenarios judged as inappropriate by the expert panel approach had cost-effectiveness ratios comparable with many generally recommended medical interventions. Formal synthesis of expert judgment and decision modeling is warranted in future efforts at guideline development.


Implementation Science and Employer Disability Practices: Embedding Implementation Factors in Research Designs.
Purpose For work disability research to have an impact on employer policies and practices it is important for such research to acknowledge and incorporate relevant aspects of the workplace. The goal of this article is to summarize recent theoretical and methodological advances in the field of Implementation Science, relate these to research of employer disability management practices, and recommend future research priorities. Methods The authors participated in a year-long collaboration culminating in an invited 3-day conference, "Improving Research of Employer Practices to Prevent Disability", held October 14-16, 2015, in Hopkinton, MA, USA. The collaboration included a topical review of the literature, group conference calls to identify key areas and challenges, drafting of initial documents, review of industry publications, and a conference presentation that included feedback from peer researchers and a question/answer session with a special panel of knowledge experts with direct employer experience. Results A 4-phase implementation model including both outer and inner contexts was adopted as the most appropriate conceptual framework, and aligned well with the set of process evaluation factors described in both the work disability prevention literature and the grey literature. Innovative interventions involving disability risk screening and psychologically-based interventions have been slow to gain traction among employers and insurers. Research recommendations to address this are : (1) to assess organizational culture and readiness for change in addition to individual factors; (2) to conduct process evaluations alongside controlled trials; (3) to analyze decision-making factors among stakeholders; and (4 ) to solicit input from employers and insurers during early phases of study design. Conclusions Future research interventions involving workplace support and involvement to prevent disability may be more feasible for implementation if organizational decision-making factors are imbedded in research designs and interventions are developed to take account of these influences.


Identification of Possible Risk Variants of Familial Strabismus Using Exome Sequencing Analysis.
To investigate candidate genes associated with familial strabismus and propose a theory of their interaction in familial strabismus associated with early neurodevelopment. Eighteen families, including 53 patients diagnosed with strabismus and 34 unaffected family members, were analyzed. All patients with strabismus and available unaffected family members were evaluated using whole exome sequencing. The primary outcome was to identify rare occurring variants among affected individuals and investigate the evidence of their genetic heterogeneity. These results were compared with exome sequencing analysis to build a comprehensive genetic profile of the study families. We observed 60 variants from 58 genes in 53 patients diagnosed with strabismus. We prioritized the most credible risk variants, which showed clear segregation in family members affected by strabismus. As a result, we found risk variants in four genes (FAT3, KCNH2, CELSR1, and TTYH1) in five families, suggesting their role in development of familial strabismus. In other families, there were several rare genetic variants in affected cases, but we did not find clear segregation pattern across family members. Genomic sequencing holds great promise in elucidating the genetic causes of strabismus; further research with larger cohorts or other related approaches are warranted.


Indocyanine green as a prospective sensitizer for photodynamic therapy of melanomas.
Spectroscopic, photochemical and biological properties of indocyanine green (ICG) are presented. Light over 800 nm is effectively absorbed by ICG. This property as well as photochemical behaviour of ICG make it a very suitable dye for photodynamic treatment of melanoma cells. Cytotoxicity of ICG itself and the effect of photodynamic therapy (PDT) were evaluated by following the growth of human (SKMEL 188) and mouse (S91) melanoma cells. The surviving fraction of the cells irradiated (lambda(ex) = 830 nm) vs non-irradiated, treated with the same dose of ICG, is significantly decreased (5- to 10-fold). These results show that ICG is a very promising dye for photodynamic therapy of melanomas.


A reconstruction method for cone-beam differential x-ray phase-contrast computed tomography.
Most existing differential phase-contrast computed tomography (DPC-CT) approaches are based on three kinds of scanning geometries, described by parallel-beam, fan-beam and cone-beam. Due to the potential of compact imaging systems with magnified spatial resolution, cone-beam DPC-CT has attracted significant interest. In this paper, we report a reconstruction method based on a back-projection filtration (BPF) algorithm for cone-beam DPC-CT. Due to the differential nature of phase contrast projections, the algorithm restrains from differentiation of the projection data prior to back-projection, unlike BPF algorithms commonly used for absorption-based CT data. This work comprises a numerical study of the algorithm and its experimental verification using a dataset measured with a three-grating interferometer and a micro-focus x-ray tube source. Moreover, the numerical simulation and experimental results demonstrate that the proposed method can deal with several classes of truncated cone-beam datasets. We believe that this feature is of particular interest for future medical cone-beam phase-contrast CT imaging applications.


Topological precision in the thalamic projection to neonatal mouse barrel cortex.
Somatosensory thalamus and cortex in rodents contain topological representations of the facial whisker pad. The thalamic representation of a single whisker ("barreloid") is presumed to project exclusively to the cortical representation ("barrel") of the same whisker; however, it was not known when this correspondence is established during early development, nor how precise the thalamocortical projection is at birth, before formation of barrels and barreloids. To answer these questions, we retrogradely labeled thalamocortical projection neurons in fixed brain slices from 0-8 d old (P0-P8) mice, by placing paired deposits of two fluorescent dyes in adjacent barrels or (before barrel formation) in adjacent loci in upper cortical layers. At all ages studied, a negligible fraction of the retrogradely labeled cells was double labeled, implying that branches of single thalamocortical axons never extended within layer IV over an area wider than a single barrel. In P0 preparations, 70% of paired dye deposits placed 75-200 microns apart resulted in statistically significant segregation of labeled cell clusters in the thalamus. Quantitative analysis indicated that on P0 about 70% of thalamocortical axons were within 1.3 presumptive barrel diameters from their topologically precise target. In P4-P8 preparations, the great majority of thalamic cells retrogradely labeled from a single barrel were found in a single barreloid, implying a 1:1 projection of barreloids to barrels. The postnatal increase in topological precision was reproduced by a computer simulation, which assumed that many aberrant axons corrected their initial targeting error by extending terminal arborizations asymmetrically, towards the center of their appropriate barrel.


Probable alcohol abuse or dependence: a risk factor for intensive-care readmission in patients undergoing elective vascular and thoracic surgical procedures. Mayo Perioperative Outcomes Group.
To determine whether alcohol abuse or dependence is a risk factor for perioperative complications, increased duration of hospital stay, and increased utilization of nursing resources in patients undergoing thoracic and vascular surgical procedures. We conducted a prospective study of all adult patients who underwent an elective vascular or thoracic surgical procedure and who received postoperative care in an intensive-care setting. Patients were screened for alcohol abuse or dependence, and actual versus expected durations of stay were evaluated. The patients' medical records were reviewed for preoperative comorbidities and perioperative complications. Fisher's exact test and the rank sum test were used in the analyses. Of 321 study subjects, 290 were classified as nonalcoholic and 31 as probable alcoholic patients. Patients in the probable alcohol abuse group had a significantly increased rate of alcohol withdrawal (12.9% versus 1.7%; P = 0.006) in comparison with patients in the nonalcoholic group. Patients in the probable alcohol abuse group were readmitted to an intensive-care unit more frequently (19.4% versus 7.9%; P = 0.047) and required sedation more often (32.3% versus 13.5%; P = 0.014) than those in the nonalcoholic group. No significant differences were found between the two study groups in intensive-care unit and hospital durations of stay or in utilization of nursing resources. A dismissal diagnosis of alcoholism was recorded for only one of four patients who had a documented withdrawal episode among those categorized in the probable alcoholic group and for three of five patients with alcohol withdrawal symptoms categorized in the nonalcoholic group. Except for the occurrence of alcohol withdrawal syndrome, study patients classified in the probable alcohol abuse group did not have more medical or surgical perioperative complications than patients in the nonalcoholic group. They did have significantly more intensive-care setting readmissions. Patients with documented alcohol withdrawal episodes frequently were dismissed without a diagnosis of substance abuse or dependence.


Increased myocardial susceptibility to repetitive ischemia with high-fat diet-induced obesity.
Obesity and diabetes are frequently associated with cardiovascular disease. When a normal heart is subjected to brief/sublethal repetitive ischemia and reperfusion (I/R), adaptive responses are activated to preserve cardiac structure and function. These responses include but are not limited to alterations in cardiac metabolism, reduced calcium responsiveness, and induction of antioxidant enzymes. In a model of ischemic cardiomyopathy inducible by brief repetitive I/R, we hypothesized that dysregulation of these adaptive responses in diet-induced obese (DIO) mice would contribute to enhanced myocardial injury. DIO C57BL/6J mice were subjected to 15 min of daily repetitive I/R while under short-acting anesthesia, a protocol that results in the development of fibrotic cardiomyopathy. Cardiac lipids and candidate gene expression were analyzed at 3 days, and histology at 5 days of repetitive I/R. Total free fatty acids (FFAs) in the cardiac extracts of DIO mice were significantly elevated, reflecting primarily the dietary fatty acid (FA) composition. Compared with lean controls, cardiac FA oxidation (FAO) capacity of DIO mice was significantly higher, concurrent with increased expression of FA metabolism gene transcripts. Following 15 min of daily repetitive I/R for 3 or 5 days, DIO mice exhibited increased susceptibility to I/R and, in contrast to lean mice, developed microinfarction, which was associated with an exaggerated inflammatory response. Repetitive I/R in DIO mice was associated with more profound significant downregulation of FA metabolism gene transcripts and elevated FFAs and triglycerides. Maladaptive metabolic changes of FA metabolism contribute to enhanced myocardial injury in diet-induced obesity.


Production trials involving use of the FAMACHA system for haemonchosis in sheep: preliminary results.
In three trials conducted on two separate farms the production of sheep treated for naturally acquired haemonchosis using the FAMACHA system of targeted selective treatment (TST) (i.e. to treat only those animals unable to manage unaided in the face of heavy Haemonchus challenge) was compared to that of suppressively drenched sheep in the same flock. As expected by the research team who developed and evaluated the FAMACHA system, TST did result in some loss in production. However, despite high levels of worm challenge in two of the trials and the fact that the comparison was with suppressive drenching which is not sustainable, the total effect was relatively small in relation to the important advantage of using the TST as regards reduced selection for anthelmintic resistance (AR). Concerning the sustainability of worm control, it is concluded that the development of drug resistance to anthelmintics leaves sheep and goat farmers in South Africa no choice but to use methods of TST such as FAMACHA. The FAMACHA system can also be a useful clinical aid for early on-farm detection of AR by farmers; the degree of improvement in the colour of the ocular mucous membrane from pale to red in individually drenched anaemic animals over a period of 7-14 days can give a good indication of the efficacy of the compound(s) used.


Association of the phenylpropanoid pathway with dormancy and adaptive trait variation in apricot (Prunus armeniaca).
Trees use many mechanisms to adapt and respond to stressful conditions. The phenylpropanoid pathway in particular is known to be associated with a diverse suite of plant stress responses. In this study, we explored the relationship between the phenylpropanoid pathway metabolite production, gene expression and adaptive trait variation associated with floral bud reactivation during and following dormancy in Prunus armeniaca L. (apricot). Concentrations of eight phenylpropanoid metabolites were measured during chill accumulation and at developmental stages corresponding to the emergence of sepals and petals in floral buds of varieties that differ phenotypically in bloom date (BD). A significant interaction effect of chill hours and BD phenotype on the concentration of each of the compounds was observed (mixed analysis of variance, P < 0.05), with the concentration of most phenylpropanoid metabolites dropping precipitously when sepals and petals emerged. While phenylpropanoid biosynthetic gene expression patterns were more variable in general, expression changed over time and was impacted, although to a lesser degree, by BD phenotype. Furthermore, separation of BD phenotypic groups was most pronounced when early and late BD varieties were at different developmental stages, i.e., 800 chill hours. Taken together, these results suggest that the phenylpropanoid pathway is associated with floral bud reactivation in apricot. Furthermore, we show that the phenylpropanoid pathway is also impacted by phenological trait variation associated with dormancy. A better understanding of how apricot and other perennial tree species respond and adapt to environmental perturbations will be critical for improvement programs aimed at identifying and breeding trees more suitable for rapidly changing environments.


Comparison of dose dependences for bioeffects of continuous-wave and high-peak power microwave emissions using gel-suspended cell cultures.
The study compared bioeffects of continuous wave (CW) microwaves and short, extremely high power pulses (EHPP) at the same carrier frequency (9.3 GHz) and average power (1.25 W). The peak transmitted power for EHPP was 250 kW (0.5-micro s pulse width, 10 p.p.s.), producing the E field of 1.57 MV/m in the waveguide. A biological endpoint was the density of yeast cells, achieved after a 6 h growth period in a solid nutrient medium (agarose gel) during EHPP or CW exposure. Owing to power losses in the medium, the specific absorption rate (SAR) ranged from 3.2 kW/kg at the exposed surface of the sample to 0.6 mW/kg at 24 mm depth. Absorption and penetration of EHPP was identical to CW, producing peak SAR values 200 000 times higher than the average SAR, as high as 650 MW/kg at the surface. CW and EHPP exposures produced highly nonuniform but identical heating patterns in exposed samples. Following the exposure, the samples were sliced in a plane perpendicular to the wave propagation, in order to separate cell masses exposed at different SAR levels. Cell density in the slices was determined by nephelometry and compared to unexposed parallel control samples. Cell density was strongly affected by irradiation, and the changes correlated well with the local temperature rise. However, the data revealed no statistically significant difference between CW and EHPP samples across the entire studied range of SAR levels (over six orders of magnitude). A trend (P<0.1) for such a difference was observed in slices that were exposed at a time average SAR of 100 W/kg and higher, which corresponded to peak SAR above 20 MW/kg for the EHPP condition. These numbers could be indicative of a threshold for a specific (not merely thermal) exposure effect if the trend is confirmed by future studies.


Sonic hedgehog is essential to foregut development.
Congenital malformation of the foregut is common in humans, with an estimated incidence of 1 in 3000 live births, although its aetiology remains largely unknown. Mice with a targeted deletion of Sonic hedgehog (Shh) have foregut defects that are apparent as early as embryonic day 9.5, when the tracheal diverticulum begins to outgrow. Homozygous Shh-null mutant mice show oesophageal atresia/stenosis, tracheo-oesophageal fistula and tracheal and lung anomalies, features similar to those observed in humans with foregut defects. The lung mesenchyme shows enhanced cell death, decreased cell proliferation and downregulation of Shh target genes. These results indicate that Shh is required for the growth and differentiation of the oesophagus, trachea and lung, and suggest that mutations in SHH and its signalling components may be involved in foregut defects in humans.


Technical feasibility and histopathologic studies of ethylene vinyl copolymer (EVAL) using a swine endovascular embolization model.
To study the safety, technical efficacy, and histopathology of the ethylene vinyl alcohol copolymer (EVAL) embolic mixture in an animal model. Microcatheterization of the rete was performed in 29 swine. The clinical, angiographic, and histopathologic consequences of superselective injection of the two principal embolic mixture components (EVAL and dimethyl sulfoxide [DMSO]) were evaluated. Necropsy and standard histologic preparation were used for pathologic analysis. Significant technical difficulties and adverse outcomes occurred. The EVAL mixture was difficult to see under fluoroscopy and prematurely polymerized during one embolization, resulting in catheter occlusion. However, polymerized EVAL did not adhere to the catheter. DMSO damaged the plastic hubs of the microcatheters. Infusions of DMSO always caused immediate, moderate to severe vasospasm and frequently caused either subarachnoid hemorrhage or stroke. Histopathologic findings of both DMSO and DMSO plus EVAL were similar, producing variable endothelial denuding, thrombosis, and disruption of internal elastic lamina in the acute stage. An intense, mixed inflammatory response, organized thrombus, and transmural necrosis with extravasation were seen in the subacute and chronic stages. Despite having some desirable features as an embolic agent, significant problems were encountered with EVAL; the most important of which is that one of the principal components of the embolic mixture, DMSO, seems to be very angiotoxic.


Improving Detection and Response to Respiratory Events - Kenya, April 2016-April 2020.
Respiratory pathogens, such as novel influenza A viruses, Middle East respiratory syndrome coronavirus (MERS-CoV), and now, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), are of particular concern because of their high transmissibility and history of global spread (1). Clusters of severe respiratory disease are challenging to investigate, especially in resource-limited settings, and disease etiology often is not well understood. In 2014, endorsed by the Group of Seven (G7),* the Global Health Security Agenda (GHSA) was established to help build country capacity to prevent, detect, and respond to infectious disease threats.† GHSA is a multinational, multisectoral collaboration to support countries towards full implementation of the World Health Organization's International Health Regulations (IHR).§ Initially, 11 technical areas for collaborator participation were identified to meet GHSA goals. CDC developed the Detection and Response to Respiratory Events (DaRRE) strategy in 2014 to enhance country capacity to identify and control respiratory disease outbreaks. DaRRE initiatives support the four of 11 GHSA technical areas that CDC focuses on: surveillance, laboratory capacity, emergency operations, and workforce development.¶ In 2016, Kenya was selected to pilot DaRRE because of its existing respiratory disease surveillance and laboratory platforms and well-developed Field Epidemiology and Laboratory Training Program (FELTP) (2). During 2016-2020, Kenya's DaRRE partners (CDC, the Kenya Ministry of Health [MoH], and Kenya's county public health officials) conceptualized, planned, and implemented key components of DaRRE. Activities were selected based on existing capacity and determined by the Kenya MoH and included 1) expansion of severe acute respiratory illness (SARI) surveillance sites; 2) piloting of community event-based surveillance; 3) expansion of laboratory diagnostic capacity; 4) training of public health practitioners in detection, investigation, and response to respiratory threats; and 5) improvement of response capacity by the national emergency operations center (EOC). Progress on DaRRE activity implementation was assessed throughout the process. This pilot in Kenya demonstrated that DaRRE can support IHR requirements and can capitalize on a country's existing resources by tailoring tools to improve public health preparedness based on countries' needs.


Comparison of monounsaturated fat with carbohydrates as a replacement for saturated fat in subjects with a high metabolic risk profile: studies in the fasting and postprandial states.
In subjects with a high prevalence of metabolic risk abnormalities, the preferred replacement for saturated fat is unresolved. The objective was to study whether carbohydrate or monounsaturated fat is a preferred replacement for saturated fat. Fifty-two men and 33 women, selected to have any combination of HDL cholesterol < or = 30th percentile, triacylglycerol > or = 70th percentile, or insulin > or = 70th percentile, were enrolled in a 3-period, 7-wk randomized crossover study. The subjects consumed an average American diet (AAD; 36% of energy from fat) and 2 additional diets in which 7% of energy from saturated fat was replaced with either carbohydrate (CHO diet) or monounsaturated fatty acids (MUFA diet). Relative to the AAD, LDL cholesterol was lower with both the CHO (-7.0%) and MUFA (-6.3%) diets, whereas the difference in HDL cholesterol was smaller during the MUFA diet (-4.3%) than during the CHO diet (-7.2%). Plasma triacylglycerols tended to be lower with the MUFA diet, but were significantly higher with the CHO diet. Although dietary lipid responses varied on the basis of baseline lipid profiles, the response to diet did not differ between subjects with or without the metabolic syndrome or with or without insulin resistance. Postprandial triacylglycerol concentrations did not differ significantly between the diets. Lipoprotein(a) concentrations increased with both the CHO (20%) and MUFA (11%) diets relative to the AAD. In the study population, who were at increased risk of coronary artery disease, MUFA provided a greater reduction in risk as a replacement for saturated fat than did carbohydrate.


On-line fluorescent monitoring of the degradation of polymeric scaffolds for tissue engineering.
Tissue engineering involves culturing, growing and assembling cells and newly generated matrix in polymeric scaffolds. To achieve a functional tissue in vitro, the cell-scaffold constructs are subjected to various stimulations during an incubation phase, which mimics the in vivo environment. In order to monitor the progression of tissue formation, there is a need for on-line and non-destructive methods of monitoring at the cellular and biomolecular level, for example, the assessment of scaffold degradation alongside the measure of matrix production. This study presents a proof of concept for monitoring scaffold degradation on-line within a culture environment. Using a mesoporous silica based approach, a pH sensitive fluorescent probe, fluorescein isothiocyanate (FITC), was incorporated into degradable polymeric scaffolds made from poly(L-lactic acid) which has a slow degradation rate, and poly(lactide-co-glycolide) which has a rapid degradation rate. The fluorescent probe was incorporated into thin films and three dimensional porous scaffolds demonstrating the capabilities of monitoring on-line. Following incubation, the intensity of fluorescence in the rapidly degrading scaffolds reduced with culture time in comparison to slow degrading polymeric scaffolds when observed qualitatively using fluorescent microscopy. The relationship between pH and fluorescent intensity was assessed, and the use of this technique for monitoring by-products via the solid scaffold by microscopy or through culture medium by a luminescence spectrometer is discussed. This study demonstrates that endowing scaffolds with a sensing element could provide an on-line and non-destructive monitoring method for tissue engineering.


Meta-analysis of Campylobacter spp. survival data within a temperature range of 0 to 42°C.
In Europe, Campylobacter is the leading reported cause of bacterial foodborne infectious disease. Quantifying its ability to survive at chilled and ambient temperatures and identifying the factors involved in variation in its survival may contribute to the development of efficient risk management strategies. A data set of 307 inactivation curves collected from the literature and the ComBase database, combined with 388 experimental curves, was analyzed with a log-linear model to obtain 695 D-values (time for 1 log inactivation). An additional 146 D-values collected from the literature or ComBase were added to the data set, for a total of 841 D-values. Because data were collected from different studies, the experimental conditions were somewhat heterogeneous (e.g., type of media or strain used). The full data set was then split into 19 different study types on which a meta-analysis was performed to determine the effect of temperature (range 0 to 42°C), Campylobacter species (C. coli and C. jejuni), and media (liquid media or meat matrix) on the survival ability of Campylobacter. A mixed-effects model, in which the study type and bacterial species were considered as random effects and the media and temperature as fixed effects, was run using a Bayesian approach. Overall, the model gave satisfactory results, with a residual standard deviation of 0.345 (the model response was the log D-value, expressed in days). In addition, the survival of Campylobacter was greater at 0 than at 42°C, with a log-linear pattern; the z-value (temperature to have a 10-fold decrease of D-value) was estimated to be 26.4°C (95 % interval: 23.9 to 29.4°C). Despite a significant media-species interaction term, it was established that both species were more resistant on the meat matrix than in liquid media. These results may be used to understand how Campylobacter can survive along the food chain, particularly in chilled environments, and consequently be transferred to other foodstuffs.


Factors Related to Not Following Up with Recommended Testing in the Diagnosis of Newborn Hearing Loss.
Children's hearing is a public health concern, and universal newborn hearing screenings are the first step in detecting and treating congenital hearing loss. Despite the high rate of participation in such programs, loss to follow-up (LTF) with additional recommended diagnosis and treatment has been a persistent problem. The current research seeks to expand the knowledge base at the point of diagnosis, where there is a large drop-off in parents following through with recommended care. This research was organized around the following question: What biopsychosocial factors are associated with LTF between screenings and diagnostic evaluations? A prospective quantitative longitudinal study tracked 203 families whose newborns were referred for additional testing at discharge from the hospital after birth. Binary logistic regression was used to determine what constellation of factors best predicted LTF. Psychosocial factors related to being lost to follow-up at diagnosis included race and ethnicity and access to health care professionals, with African American babies being most at risk for LTF; however, the impact of race and ethnicity declined when parents believed they had more health care professionals with whom to consult.


Practice parameter: the neurodiagnostic evaluation of the child with a first simple febrile seizure. American Academy of Pediatrics. Provisional Committee on Quality Improvement, Subcommittee on Febrile Seizures.
The American Academy of Pediatrics and its Provisional Committee on Quality Improvement, in collaboration with experts from the Section on Neurology, general pediatricians, consultants in the fields of neurology and epilepsy, and research methodologists, developed this practice parameter. This parameter provides recommendations for the neurodiagnostic evaluation of a child with a first simple febrile seizure. These recommendations derive from both a thorough review of the literature and expert consensus. Interventions of direct interest include lumbar puncture, electroencephalography, blood studies, and neuroimaging. The methods and results of the literature review and data analyses can be found in the technical report that is available from the Publications Department of the American Academy of Pediatrics. This parameter is designed to assist pediatricians by providing an analytic framework for the evaluation and treatment of this condition. It is not intended to replace clinical judgement or establish a protocol for all patients with this condition. It rarely will be the only appropriate approach to the problem.


Dyscoordination of non-rapid eye movement sleep oscillations in autism spectrum disorder.
Converging evidence from neuroimaging, sleep, and genetic studies suggest that dysregulation of thalamocortical interactions mediated by the thalamic reticular nucleus (TRN) contribute to autism spectrum disorder (ASD). Sleep spindles assay TRN function, and their coordination with cortical slow oscillations (SOs) indexes thalamocortical communication. These oscillations mediate memory consolidation during sleep. In the present study, we comprehensively characterized spindles and their coordination with SOs in relation to memory and age in children with ASD. Nineteen children and adolescents with ASD, without intellectual disability, and 18 typically developing (TD) peers, aged 9-17, completed a home polysomnography study with testing on a spatial memory task before and after sleep. Spindles, SOs, and their coordination were characterized during stages 2 (N2) and 3 (N3) non-rapid eye movement sleep. ASD participants showed disrupted SO-spindle coordination during N2 sleep. Spindles peaked later in SO upstates and their timing was less consistent. They also showed a spindle density (#/min) deficit during N3 sleep. Both groups showed significant sleep-dependent memory consolidation, but their relations with spindle density differed. While TD participants showed the expected positive correlations, ASD participants showed the opposite. The disrupted SO-spindle coordination and spindle deficit provide further evidence of abnormal thalamocortical interactions and TRN dysfunction in ASD. The inverse relations of spindle density with memory suggest a different function for spindles in ASD than TD. We propose that abnormal sleep oscillations reflect genetically mediated disruptions of TRN-dependent thalamocortical circuit development that contribute to the manifestations of ASD and are potentially treatable.


Adsorption behavior of 4-methoxypyridine on gold nanoparticles.
We demonstrate a phase transfer method to create stable colloidal solutions of Au nanoparticles with 4-methoxypyridine ligands. We then investigate the adsorption behavior of 4-methoxypyridine onto gold surfaces by Raman spectroscopy, DFT calculations, and (1)H NMR. In contrast to unsubstituted pyridine and the frequently used (N,N-dimethylamino)pyridine (DMAP), a flat adsorption of 4-methoxypyridine on gold was found.


Physical and sexual abuse among homeless and unstably housed adults living with HIV: prevalence and associated risks.
We examined the prevalence and risks associated with interpersonal (physical and sexual) abuse among HIV-seropositive homeless or unstably housed adults. Data were obtained from the Housing and Health Study of participants living in Baltimore, Chicago, and Los Angeles (n = 644). We used logistic regression to identify risks associated with abuse. About 77% of men and 86% of women reported ever experiencing abuse. Women were at greater risk than men for intimate partner physical abuse, childhood sexual abuse (CSA), and adulthood sexual abuse. Men and women experiencing intimate partner physical abuse reported increased risk of unprotected sex. Other risks associated with abuse include sex exchange; lifetime alcohol abuse; and depressive symptoms. Abuse prevalence among sample exceeds those found in other samples of general USA, HIV-seropositive, and homeless populations. Identifying persons at risk of abuse is needed to reduce risk among homeless or unstably housed persons living with HIV.


Reconstruction of the human visual system based on DTI fiber tracking.
To apply and to evaluate the newly developed advanced fast marching algorithm (aFM) in vivo by reconstructing the human visual pathway, which is characterized by areas of extensive fiber crossing and branching, i.e., the optic chiasm and the lateral geniculate nucleus (LGN). Diffusion tensor images were acquired in 10 healthy volunteers. Due to the proximity to bony structures and air-filled spaces of the optic chiasm, a high sensitivity encoding (SENSE) reduction factor was applied to reduce image distortions in this area. To reconstruct the visual system, three different seed areas were chosen separately. The results obtained by the aFM tracking algorithm were compared and validated with known anatomy. The visual system could be reconstructed reproducibly in all subjects and the reconstructed fiber pathways are in good agreement with known anatomy. The present work shows that the advanced aFM, which is especially designed for overcoming tracking limitations within areas of extensive fiber crossing, handles the fiber crossing and branching within the optic chiasm and the LGN correctly, thus allowing the reconstruction of the entire human visual fiber pathway, from the intraorbital segment of the optic nerves to the visual cortex.


The spectrum of acute bacterial meningitis in elderly patients.
We conducted a prospective, observational study in Barcelona to determine the epidemiology, clinical features, and outcome of elderly patients with acute bacterial meningitis (ABM) compared with younger adults. During 1982-2010, all patients with ABM were prospectively evaluated. There were two groups: I (15-64 years) and II (≥ 65 years). All patients underwent clinical examination on admission and at discharge following a predefined protocol. We evaluated 635 episodes of ABM. The incidence was 4.03/100,000 (Group I) and 7.40 /100,000 inhabitants/year (Group II) (RR = 1.84; 95%CI: 1.56-2.17, P < 0.0001). Elderly patients had co-morbid conditions more frequently (P < 0.0001) and more frequently lacked fever (P = 0.0625), neck stiffness (P < 0.0001) and skin rash (P < 0.0001), but had an altered level of consciousness more often (P < 0.0001). The interval admission-start of antibiotic therapy was longer for elderly patients (P < 0.0001). Meningococcal meningitis was less frequent in elderly patients (P < 0.0001), whereas listerial (P = 0.0196), gram-negative bacillary (P = 0.0065), and meningitis of unknown origin (P = 0.0076) were more frequent. Elderly patients had a higher number of neurologic (P = 0.0009) and extra-neurologic complications (P < 0.0001). The overall mortality ratio was higher in elderly patients (P < 0.0001). Elderly people are at higher risk of having ABM than younger adults. ABM in the elderly presents with co-morbid conditions, is clinically subtler, has a longer interval admission-antibiotic therapy, and has non-meningococcal etiology. It is associated with an earlier and higher mortality rate than in younger patients.


Plant development: pulled up by the roots.
Recent advances in the study of root development in Arabidopsis have begun to yield mechanistic insights into the processes that underpin morphogenesis, pattern formation and cell differentiation in plants. The most exciting feature of these advances is that, as a consequence of the simple and largely invariant cellular architecture of the root, the processes can be studied at a cellular level. Nevertheless, it is clear that although the cell lineages of the root are relatively invariant, we have no evidence that lineage per se is an important regulator of development. Instead, all the evidence indicates that the positional regulation of inductive cues is of primary importance. The availability of new root mutants is alerting us to the complexity of the contribution of cell size and cell expansion to plant development.


Sedation in the intensive care unit.
To describe the goals of sedative use in the intensive care unit and review the pharmacology of commonly used sedative drugs as well as to review pertinent publications in the literature concerning the comparative pharmacology of these drugs, with emphasis on outcomes related to sedation and comparative pharmacoeconomics. Publications in the scientific literature. Computer search of the literature with selection of representative articles. Proper choice and use of sedative drugs is based on knowledge of the pharmacology of commonly used agents and is an essential component of caring for patients in the intensive care unit. The large variability in pharmacokinetics and pharmacodynamics in the critically ill make it difficult to directly compare agents. Midazolam provides rapid and reliable amnesia, even when administered for low levels of sedation. Propofol may be useful when deeper levels of sedation and more rapid awakening are required. Lorazepam can be used for long-term sedation in more stable patients if rapidity of effect is not required. Further investigation in assessment of depth of sedation in the critically ill is needed. Continued study of costs, side effects, and appropriate dosing strategies of all sedative agents is needed to answer questions not sufficiently addressed in the current literature. An individualized approach to sedation based on knowledge of drug pharmacology is needed because of confounding variables including concurrent patient illness, depth of sedation, and concomitant use of analgesic agents. (Crit Care Med 2000; 28:854-866)


Mechanism of the tissue distribution and biliary excretion of the cyclic peptide octreotide.
The hepatobiliary transport and tissue distribution of the cationic cyclooctapeptide octreotide were studies at steady state after its infusion, at various rates, in rats. After an increase in steady-state plasma concentration, marked decrease in the tissue to plasma concentration ratio was observed only in pancreas, the target organ of octreotide. A marked decrease in the biliary excretion clearance, defined with respect to the concentration in the liver, was also observed, suggesting that a transport carrier was involved in the biliary excretion. The plasma elimination and biliary excretion profiles of octreotide were determined in Eisai hyperbilirubinemic rats (EHBR), which have an hereditary defect of the active transport carrier for organic anions in bile canalicular membranes. Although biliary excretion of octreotide was significantly reduced in EHBR, compared with normal Sprague-Dawley rats, no difference was observed in biliary excretion clearance, defined with respect to the concentration in the liver, between Sprague-Dawley rats and EHBR. On the other hand, the liver to plasma concentration ratio in EHBR fell to half that in Sprague-Dawley rats. These results suggest that the decreased biliary excretion of octreotide in EHBR is due not to reduced biliary excretion ability but to reduced hepatic uptake of octreotide. We studied in vitro transport using bile canalicular membrane vesicles. A significant increase in the transport of octreotide by bile canalicular membrane vesicles was observed in the presence of ATP, and the estimated kinetic parameters K(m) and Vmax were 6.5 microM and 370 pmol/min/mg of protein, respectively. Similar ATP-dependent uptake was observed in bile canalicular membrane vesicles prepared from EHBR. We concluded that the biliary excretion of octreotide is by ATP-dependent primary active transport and that the carrier system for octreotide differs from the so-called "canalicular multispecific organic anion transporter," which is absent in EHBR.


Serial expression analysis of breast tumors during neoadjuvant chemotherapy reveals changes in cell cycle and immune pathways associated with recurrence and response.
The molecular biology involving neoadjuvant chemotherapy (NAC) response is poorly understood. To elucidate the impact of NAC on the breast cancer transcriptome and its association with clinical outcome, we analyzed gene expression data derived from serial tumor samples of patients with breast cancer who received NAC in the I-SPY 1 TRIAL. Expression data were collected before treatment (T1), 24-96 hours after initiation of chemotherapy (T2) and at surgery (TS). Expression levels between T1 and T2 (T1 vs. T2; n = 36) and between T1 and TS (T1 vs. TS; n = 39) were compared. Subtype was assigned using the PAM50 gene signature. Differences in early gene expression changes (T2 - T1) between responders and nonresponders, as defined by residual cancer burden, were evaluated. Cox proportional hazards modeling was used to identify genes in residual tumors associated with recurrence-free survival (RFS). Pathway analysis was performed with Ingenuity software. When we compared expression profiles at T1 vs. T2 and at T1 vs. TS, we detected significantly altered expression of 150 and 59 transcripts, respectively. We observed notable downregulation of proliferation and immune-related genes at T2. Lower concordance in subtype assignment was observed between T1 and TS (62 %) than between T1 and T2 (75 %). Analysis of early gene expression changes (T2 - T1) revealed that decreased expression of cell cycle inhibitors was associated with poor response. Increased interferon signaling (TS - T1) and high expression of cell proliferation genes in residual tumors (TS) were associated with reduced RFS. Serial gene expression analysis revealed candidate immune and proliferation pathways associated with response and recurrence. Larger studies incorporating the approach described here are warranted to identify predictive and prognostic biomarkers in the NAC setting for specific targeted therapies. ClinicalTrials.gov identifier: NCT00033397 . Registered 9 Apr 2002.


Chronic Pain in Chronic Heart Failure: A Review Article.
Heart failure (HF) is one of the main causes of death and disability in the world. The prevalence of HF in developed countries is between 1% and 2% of the adult population and approximately between 6% and 10% in the elderly, giving rise to high costs of care and treatment. Indeed, in the United States, the direct and indirect costs exceeded 23 billion dollars in 2002. HF is typically characterized by periods of acute symptoms followed by returns to nearly asymptomatic periods. As dyspnea and fatigue are considered the signature symptoms of HF, other symptoms such as pain go unnoticed. Awareness of the burden of pain, however, is growing in patients with chronic HF. The past 2 decades have witnessed remarkable technical headway in cardiology and many patients have survived despite the progressive impairment of their cardiovascular function. It is, therefore, of great value to investigate the prevalence and management of pain in patients with HF. To that end, we undertook a comprehensive search using the MEDLINE database for studies and guidelines on the subject of pain and HF and the complications and considerations and finally selected 65 studies for review.


Risk factors for developing osteomyelitis in patients with diabetic foot wounds.
Osteomyelitis worsens the prognosis in the diabetic foot, but predisposing factors remain largely undefined. In a prospectively followed cohort we assessed risk factors for developing osteomyelitis. We enrolled consecutive persons with diabetes who presented to a managed-care diabetes disease management program. The patients underwent standardized assessments. We monitored for all foot complications, defined infections by criteria consistent with International Working Group guidelines, and defined osteomyelitis as a positive culture from a bone specimen. 1666 persons were enrolled, 50% male, mean age 69 years. Over a mean of 27.2 months of follow-up, 151 patients developed foot infections, 30 (19.9%) of which involved bone. Independent risk factors for osteomyelitis were: wounds that extended to bone or joint (relative risk [RR]=23.1), previous history of a wound prior to enrollment (RR=2.2), and recurrent or multiple wounds during the study period (RR=1.9). In this study population, managed in a specialized diabetic foot care center, the results suggest that independent risk factors for developing osteomyelitis are deep, recurrent and multiple wounds. These results may help clinicians target their efforts at diagnosing foot osteomyelitis to the highest risk patients.


Hyperspectral Analysis of Leaf Pigments and Nutritional Elements in Tallgrass Prairie Vegetation.
Understanding the spatial distribution of forage quality is important to address critical research questions in grassland science. Due to its efficiency and accuracy, there has been a widespread interest in mapping the canopy vegetation characteristics using remote sensing methods. In this study, foliar chlorophylls, carotenoids, and nutritional elements across multiple tallgrass prairie functional groups were quantified at the leaf level using hyperspectral analysis in the region of 470-800 nm, which was expected to be a precursor to further remote sensing of canopy vegetation quality. A method of spectral standardization was developed using a form of the normalized difference, which proved feasible to reduce the interference from background effects in the leaf reflectance measurements. Chlorophylls and carotenoids were retrieved through inverting the physical model PROSPECT 5. The foliar nutritional elements were modeled empirically. Partial least squares regression was used to build the linkages between the high-dimensional spectral predictor variables and the foliar biochemical contents. Results showed that the retrieval of leaf biochemistry through hyperspectral analysis can be accurate and robust across different tallgrass prairie functional groups. In addition, correlations were found between the leaf pigments and nutritional elements. Results provided insight into the use of pigment-related vegetation indices as the proxy of plant nutrition quality.