Expression of the cystic fibrosis gene in adult human lung.
Critical to an understanding of the pulmonary disease in cystic fibrosis (CF) and the development of effective gene therapies is a definition of the distribution and regulation of CF gene expression in adult human lung. Previous studies have detected the product of the CF gene, the CF transmembrane conductance regulator (CFTR), in submucosal glands of human bronchi. In this report, we have characterized the distribution of CFTR RNA and protein in the distal airway and alveoli of human lungs. Samples from eight human lungs were analyzed for CFTR expression by in situ hybridization and immunocytochemistry. CFTR was detected in a subpopulation of epithelial cells at every level of the distal lung, including proximal, terminal, and respiratory bronchioles, and the alveoli. However, there was substantial variation in the level of CFTR expression between samples. In bronchioles, CFTR protein localized to the apical plasma membrane and was found primarily in a subpopulation of nonciliated cells. CFTR was expressed in the same distribution as the Clara cell marker CC10 in proximal bronchioles, however, expression was discordant in the more distal bronchioles and alveoli where CC10 was not detected. These studies suggest that epithelial cells of the distal lung may play a primary role in the pathogenesis of CF as well as expand the spectrum of target cells that should be considered in the development of gene therapies.


High-resolution whole-body magnetic resonance image tumor staging with the use of parallel imaging versus dual-modality positron emission tomography-computed tomography: experience on a 32-channel system.
The objective of this study was to compare the accuracy in staging of various malignant tumors with whole-body magnetic resonance imaging (WB-MRI) using parallel imaging (PAT) and positron emission tomography-computed tomography (PET-CT). In a prospective study, 41 patients withoncologic diseases underwent [F]-fluoro-2-deoxy-D-glucose PET-CT for tumor staging and WB-MRI on a 32-channel-scanner with the use of PAT. Coronal T1w and STIR sequences at 5 body levels, axial HASTE imaging of the lung, and contrast-enhanced T1w sequences of the liver, brain, and abdomen were performed. TNM stage was assessed for both modalities in a separate consensus reading using histologic results and radiologic follow up within 6 months as the standard of reference. Three primary and 4 recurrent tumors were detected; one recurrent tumor was missed with WB-MRI. Sixty benign and 60 malignant lymph nodes were detected with a sensitivity of 98% and specificity of 83% for PET-CT and 80%/75% for WB-MRI, respectively. One hundred ninety-one malignant and 77 benign distant lesions were detected with a sensitivity/specificity of 82% for PET-CT and 96%/82% for WB-MRI. Accuracy for correct TNM staging was 96% for PET-CT and 91% for WB-MRI. WB-MRI and PET-CT are reliable imaging modalities for tumor staging. WB-MRI is highly sensitive in detecting distant metastases; PET-CT is superior in lymph node staging. PAT makes high-resolution WB-MRI feasible within less than 1 hour.


Lifetime Prediction of Polymers: To Bet, or Not to Bet-Is This the Question?
Polymers are a great and very important category of organic compounds that have changed our lifestyle. In the last eighty years, we have used them for the most varied applications, and from the first structural ones we began to investigate their durability, which can be fatal in the successful completion of the application for which the material was designed. Over the last thirty years, the environmental problems related to the disposal of polymers that have completed their lifecycle have begun to arise, and the need to foresee their end of life has become increasingly urgent. In this manuscript, the reliability of the lifetime predictions of polymeric materials is faced with comparing measurements obtained at low temperature with those carried out at high temperatures, in the molten state. The obtained data were treated by a well-established kinetics model and discrepancies were observed in the two different conditions (high and low temperatures), which led to a mismatching between expected and real data. A correction of the data extrapolated from measurements obtained at high temperatures, by using a novel equation which takes into account the induction period (IP) of the degradation process, is proposed. Considerations about the useful parameters, namely initial decomposition temperature (Ti), activation energy of degradation (Ea), and glass-transition temperature (Tg), to be used for making predictions, are also carried out.


DNA sequences from multiple amplifications reveal artifacts induced by cytosine deamination in ancient DNA.
We show that DNA molecules amplified by PCR from DNA extracted from animal bones and teeth that vary in age between 25 000 and over 50 000 years carry C-->T and G-->A substitutions. These substitutions can reach high proportions among the molecules amplified and are due to the occurrence of modified deoxycytidine residues in the template DNA. If the template DNA is treated with uracil N-glycosylase, these substitutions are dramatically reduced. They are thus likely to result from deamination of deoxycytidine residues. In addition, 'jumping PCR', i.e. the occurrence of template switching during PCR, may contribute to these substitutions. When DNA sequences are amplified from ancient DNA extracts where few template molecules initiate the PCR, precautions such as DNA sequence determination of multiple clones derived from more than one independent amplification are necessary in order to reduce the risk of determination of incorrect DNA sequences. When such precautionary measures are taken, errors induced by damage to the DNA template are unlikely to be more frequent than approximately 0.1% even under the unlikely scenario where each amplification starts from a single template molecule.


Retinoic acid drives aryl hydrocarbon receptor expression and is instrumental to dioxin-induced toxicity during palate development.
Palate development depends on complex events and is very sensitive to disruption. Accordingly, clefts are the most common congenital malformations worldwide, and a connection is proposed with fetal exposure to toxic factors or environmental contaminants, such as dioxins. There is increasing evidence that dioxin interferes with all-trans-retinoic acid (atRA), a hormone-like signal derived from vitamin A, which plays an essential role during embryonic development. Although similarities have been described between dioxin-induced toxicity and the outcome of altered atRA signaling during palate development, their relationship needs to be clarified. We used a genetic approach to understand the interaction between atRA and dioxin and to identify the cell type targeted by dioxin toxicity during secondary palate formation in mice. We analyzed the phenotype of mouse embryos harboring an atRA-sensitive reporter transgene or bearing null mutations for atRA-synthesizing enzymes (RALDH) or atRA receptors (RAR) and maternally exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) at gestation day 10.5. We found that an intact atRA signal was required to enable TCDD to induce cleft palate. This mandatory atRA signal was generated through the activity of RALDH3 in the nasal epithelium and was transduced by RARγ (RARG) in the nasal mesenchyme, where it notably controlled aryl hydrocarbon receptor (Ahr) transcript levels. TCDD also did not alter the developmental pattern of atRA signaling during palate formation. TCDD-induced alteration of secondary palate development in the mouse appears to depend on atRA signaling, which controls AHR expression. This mechanism is likely conserved throughout vertebrate evolution and may therefore be relevant in humans.


Evaluation of the Response of HNSCC Cell Lines to γ-Rays and 12C Ions: Can Radioresistant Tumors Be Identified and Selected for 12C Ion Radiotherapy?
Head and neck squamous cell carcinoma (HNSCC) is the sixth most common malignancy worldwide. Thirty percent of patients will experience locoregional recurrence for which median survival is less than 1 year. Factors contributing to treatment failure include inherent resistance to X-rays and chemotherapy, hypoxia, epithelial to mesenchymal transition, and immune suppression. The unique properties of 12C radiotherapy including enhanced cell killing, a decreased oxygen enhancement ratio, generation of complex DNA damage, and the potential to overcome immune suppression make its application well suited to the treatment of HNSCC. We examined the 12C radioresponse of five HNSCC cell lines, whose surviving fraction at 3.5 Gy ranged from average to resistant when compared with a larger panel of 38 cell lines to determine if 12C irradiation can overcome X-ray radioresistance and to identify biomarkers predictive of 12C radioresponse. Cells were irradiated with 12C using a SOBP with an average LET of 80 keV/μm (CNAO: Pavia, Italy). RBE values varied depending upon endpoint used. A 37 gene signature was able to place cells in their respective radiosensitivity cohort with an accuracy of 86%. Radioresistant cells were characterized by an enrichment of genes associated with radioresistance and survival mechanisms including but not limited to G2/M Checkpoint MTORC1, HIF1α, and PI3K/AKT/MTOR signaling. These data were used in conjunction with an in silico-based modeling approach to evaluate tumor control probability after 12C irradiation that compared clinically used treatment schedules with fixed RBE values vs. the RBEs determined for each cell line. Based on the above analysis, we present the framework of a strategy to utilize biological markers to predict which HNSCC patients would benefit the most from 12C radiotherapy.


Eribulin in Male Patients With Breast Cancer: The First Report of Clinical Outcomes.
Evidence on the management and treatment of male breast cancer is scant. We report the analysis of a multicenter Italian series of patients with male breast cancer treated with eribulin. To our knowledge, this is the first report on the use or eribulin in this setting. Patients were retrospectively identified in 19 reference centers. All patients received eribulin treatment, according to the standard practice of each center. Data on the identified patients were collected using a standardized form and were then centrally reviewed by two experienced oncologists. A total of 23 patients (median age, 64 years; range, 42-80) were considered. The median age at the time of diagnosis of breast cancer was 57 years (range, 42-74). HER2 status was negative in 14 patients (61%), and 2 patients (9%) had triple-negative disease. The most common metastatic sites were the lung (n = 14; 61%) and bone (n = 13; 56%). Eribulin was administered for a median of 6 cycles (range, 3-15). All patients reported at least stable disease; two complete responses (9%) were documented. Eribulin was well-tolerated, with only four patients (17%) reporting grade 3 adverse events and two (9%) with treatment interruptions because of toxicity. Eight subjects (35%) did not report any adverse event during treatment. For patients with a reported fatal event, the median overall survival from the diagnosis of metastatic disease was 65 months (range, 22-228). Although hampered by all the limitations of any retrospective case series, the results of the present study suggest, for the first time, the use of eribulin as therapy for male breast cancer. Evidence on the management and treatment of male breast cancer is eagerly awaited. Although hampered by all the limitations of any retrospective case series, the results of the present study suggest, for the first time, the use of eribulin as therapy for male breast cancer.


Activation of PKR causes amyloid ß-peptide accumulation via de-repression of BACE1 expression.
BACE1 is a key enzyme involved in the production of amyloid ß-peptide (Aß) in Alzheimer's disease (AD) brains. Normally, its expression is constitutively inhibited due to the presence of the 5'untranslated region (5'UTR) in the BACE1 promoter. BACE1 expression is activated by phosphorylation of the eukaryotic initiation factor (eIF)2-alpha, which reverses the inhibitory effect exerted by BACE1 5'UTR. There are four kinases associated with different types of stress that could phosphorylate eIF2-alpha. Here we focus on the double-stranded (ds) RNA-activated protein kinase (PKR). PKR is activated during viral infection, including that of herpes simplex virus type 1 (HSV1), a virus suggested to be implicated in the development of AD, acting when present in brains of carriers of the type 4 allele of the apolipoprotein E gene. HSV1 is a dsDNA virus but it has genes on both strands of the genome, and from these genes complementary RNA molecules are transcribed. These could activate BACE1 expression by the PKR pathway. Here we demonstrate in HSV1-infected neuroblastoma cells, and in peripheral nervous tissue from HSV1-infected mice, that HSV1 activates PKR. Cloning BACE1 5'UTR upstream of a luciferase (luc) gene confirmed its inhibitory effect, which can be prevented by salubrinal, an inhibitor of the eIF2-alpha phosphatase PP1c. Treatment with the dsRNA analog poly (I∶C) mimicked the stimulatory effect exerted by salubrinal over BACE1 translation in the 5'UTR-luc construct and increased Aß production in HEK-APPsw cells. Summarizing, our data suggest that PKR activated in brain by HSV1 could play an important role in the development of AD.


Elimination of the Perinatal Transmission of HIV and Syphilis in Puerto Rico and Sustained Success since 2007: Convergence of Science, Women-Centered Care, and Policy.
There have been significant successes in the fight against HIV/AIDS due to the access to rapid HIV testing, interventions to reduce the mother-to-child transmission (MTCT) risk, potent and effective antiviral medications, and other biomedical prevention strategies. The purpose of this work is to demonstrate that Puerto Rico eliminated Mother-to-Child Transmission of HIV (MTCT) following the 2017 World Health Organization (WHO) criteria for validating the elimination of MTCT and Syphilis. Existing epidemiological data from Puerto Rico was used to document the elimination of MTCT and Syphilis. Data to calculate the indicators was obtained from the various divisions of the Puerto Rico Department of Health, including vital statistics, surveillance data, and programmatic outcomes. Puerto Rico eliminated MTCT and syphilis, according to the WHO indicators, earlier than other countries. We can trace the outcomes to 1994 using the incidence rate of perinatally-acquired HIV of <50/100,000; to 2007 using HIV perinatal transmission rates for non-breastfeeding countries (<2%), to 2008 using 90% of women receiving ART at delivery, and to 2005 using the incidence rate of congenital syphilis of <50/100,000. Not only have we eliminated the MTCT of HIV and syphilis, but the efforts have been sustained since 2000. The elimination of transmission of infectious diseases requires the intersection of scientific feasibility, coordinated interventions, and political will, successfully attained in Puerto Rico.


The long-term mental health risk associated with non-heterosexual orientation.
Findings that describe the mental health risk associated with non-heterosexual orientation in young and middle-aged adults are from cross-sectional designs or fail to discriminate homosexual and bisexual orientations. This study examines the mental health risk of homosexual and bisexual orientation over an 8-year period. Participants were from the age-cohort study, the Personality and Total Health Through Life Project, were observed twice every 4 years, and aged 20-24 (n = 2353) and 40-44 (n = 2499) at baseline. Homosexual orientation was unrelated to long-term depression risk. Risk for anxiety and depression associated with homosexual and bisexual orientations, respectively, were attenuated in fully-adjusted models. Bisexual orientation risk associated with anxiety was partially attenuated in fully-adjusted models. Non-heterosexual orientation was not a major risk factor for long-term mental health outcomes. Instead, those with a non-heterosexual orientation were more likely to experience other mental health risk factors, which explain most of the risk observed amongst those with a non-heterosexual orientation.


AMPK: A bridge between diabetes mellitus and Alzheimer's disease.
The pathogenesis and etiology of diabetes mellitus (DM) and Alzheimer's disease (AD) share many common cellular and molecular themes. Recently, a growing body of research has shown that AMP-activated protein kinase (AMPK), a biomolecule that regulates energy balance and glucose and lipid metabolism, plays key roles in DM and AD. In this review, we summarize the relevant research on the roles of AMPK in DM and AD, including its functions in gluconeogenesis and insulin resistance (IR) and its relationships with amyloid β-protein (Aβ), Tau and AMPK activators. In DM, AMPK is involved in the regulation of glucose metabolism and IR. AMPK is closely related to gluconeogenesis, which can not only be activated by the upstream kinases liver kinase B1 (LKB1), transforming growth factor β-activated kinase 1 (TAK1), and Ca2+/calmodulin-dependent protein kinase kinase β (CaMKKβ) but also regulate the downstream kinases glucose-6-phosphatase (G-6-Pase) and phosphoenolpyruvate carboxy kinase (PEPCK), thereby affecting gluconeogenesis and ameliorating DM. Moreover, AMPK can regulate glucose transporter 4 (GLUT4) and free fatty acids to improve IR. In AD, AMPK can ameliorate abnormal brain energy metabolism, not only by reduces Aβ deposition through β-secretase but also reduces tau hyperphosphorylation through sirtuin 1 (SIRT1) and protein phosphatase 2A (PP2A). Therefore, AMPK is a bridge between DM and AD.


Anti-host isohemagglutinin production is associated with a higher risk of acute GVHD in ABO-incompatible transplantation.
We retrospectively analyzed the association between anti-host isohemagglutinin (IH) production and the development of acute GVHD. Of 189 patients who received minor or major/minor ABO-incompatible hematopoietic SCT (HSCT) at our hospital, 36 patients (19%) showed IH production. IH was detected before the onset of acute GVHD in 10, around the same time in 8, and after the onset of acute GVHD in 17 patients. The cumulative incidence of grade II-IV acute GVHD was significantly higher in the IH+ group compared with the IH- group (P<0.0001). The higher risk of acute GVHD that was associated with IH production occurred irrespective of human leukocyte Ag compatibility and donor type. Furthermore, the incidence of acute GVHD in the IH- group was comparable to that seen in major ABO-incompatible or -compatible HSCT. Our findings not only showed a strong association between IH production and acute GVHD development, but also suggested that IH production might be a useful predictor of subsequent acute GVHD after ABO-incompatible HSCT.


Caveats for the spatial arrangement method: Comment on Hout, Goldinger, and Ferguson (2013).
The gold standard among proximity data collection methods for multidimensional scaling is the (dis)similarity rating of pairwise presented stimuli. A drawback of the pairwise method is its lengthy duration, which may cause participants to change their strategy over time, become fatigued, or disengage altogether. Hout, Goldinger, and Ferguson (2013) recently made a case for the Spatial Arrangement Method (SpAM) as an alternative to the pairwise method, arguing that it is faster and more engaging. SpAM invites participants to directly arrange stimuli on a computer screen such that the interstimuli distances are proportional to psychological proximity. Based on a reanalysis of the Hout et al. (2013), data we identify three caveats for SpAM. An investigation of the distributional characteristics of the SpAM proximity data reveals that the spatial nature of SpAM imposes structure on the data, invoking a bias against featural representations. Individual-differences scaling of the SpAM proximity data reveals that the two-dimensional nature of SpAM allows individuals to only communicate two dimensions of variation among stimuli properly, invoking a bias against high-dimensional scaling representations. Monte Carlo simulations indicate that in order to obtain reliable estimates of the group average, SpAM requires more individuals to be tested. We conclude with an overview of considerations that can inform the choice between SpAM and the pairwise method and offer suggestions on how to overcome their respective limitations.


Specific interaction of type I receptors of the TGF-beta family with the immunophilin FKBP-12.
Transforming growth factor-beta (TGF-beta) family members bind to receptors that consist of heteromeric serine-threonine kinase subunits (type I and type II). In a yeast genetic screen, the immunophilin FKBP-12, a target of the macrolides FK506 and rapamycin, interacted with the type I receptor for TGF-beta and with other type I receptors. Deletion, point mutation, and co-immunoprecipitation studies further demonstrated the specificity of the interaction. Excess FK506 competed with type I receptors for binding to FKBP-12, which suggests that these receptors share or overlap the macrolide binding site on FKBP-12, and therefore they may represent its natural ligand. The specific interaction between the type I receptors and FKBP-12 suggests that FKBP-12 may play a role in type I receptor-mediated signaling.


Fibromyalgia 2016 criteria and assessments: comprehensive validation in a Norwegian population.
Background and aims The ACR1990 criteria of fibromyalgia (FM) have been criticized due to poor reliability of tender points counting (TPC), inconsistent definitions of the widespread pain, and by not considering other symptoms than pain in the FM phenotype. Therefore, several newer self-report measures for FM criteria have emerged. The aim of this study was to translate the fibromyalgia survey questionnaire (FSQ) to Norwegian and validate both the 2011 and the 2016 fibromyalgia survey diagnostic criteria (FSDC) against the ACR1990 criteria. Methods One hundred and twenty chronic pain patients formerly diagnosed with fibromyalgia according to the ACR1990 criteria, and 62 controls not diagnosed or where fibromyalgia was not suspected, were enrolled in this study. All responded to a Norwegian version of the FSQ. Also, they had a clinical examination according to ACR1990 fibromyalgia criteria including a counting of significant tender points with an algometer (TPC). The FSQ with the Widespread Pain Index (WPI) and Symptom Severity scale (SSS) subscales, Fibromyalgia Severity (FS) sum score, was examined for correlations with the fibromyalgia impact questionnaire (FIQ) and TPCs. Face-validity, internal consistence, test-retest reliability and construct validity with convergent and divergent approaches were examined and a Receiver Operating Characteristics (ROC) analysis was performed. Results The internal consistency of FS measured by Cronbach's alfa was good (=0.904). The test-retest reliability measures using intra class correlation were respectable for the FS, including WPI and SSS subscales (0.86, 0.84 and 0.87). FS, WPI and SSS correlated significantly with FIQ (0.74, 0.59 and 0.85) and TPC indicating an adequate construct, convergent validity. The medians of FS, WPI and SSS in the fibromyalgia-group were significantly different from the non-fibromyalgia-group indicating good construct, divergent validity. Using the 2011 and 2016 FSDC vs. ACR 1990 as a reference, sensitivity, specificity, positive likelihood ratio (LR +) and negative likelihood ratio (LR-) were identified. The accuracy rate for both 2011 and 2016 FSDC were respectable (84%). ROC analysis using FS revealed a very good Area Under the Curve (AUC) = 0.860. Conclusion The current study revealed that the Norwegian versions of FSQ is a valid tool for assessment of fibromyalgia according to the 2011 and 2016 (FSDC).


Alterations in calcium metabolism in murine macrophages by the benzene metabolite 1,4-benzoquinone.
Exposure of murine peritoneal macrophages to very low concentrations of 1,4-benzoquinone (BQ) induced immediate increases in intracellular Ca2+ concentrations ([Ca2+]i). Increases in [Ca2+]i were induced by concentrations as low as 5 nM and the response was dose dependent and linear up to 1 microM. The sources of Ca2+ were from both internal inositol triphosphate (IP3)-sensitive and -insensitive sites and from the external medium. BQ did not induce IP3 formation and did not affect binding to its receptors. 1, 4-Hydroquinone had no effect on [Ca2+]i. Catechol did elicit some increases in [Ca2+]i, but did so only at much higher concentrations (5 microM). The action of BQ was almost identical to that of the established Ca2+-ATPase inhibitor thapsigargin except that there were some intracellular stores of Ca2+ released by thapsigargin that were not released by BQ. BQ also was mitogenic for macrophages in conjunction with phorbol myristate acetate. These data suggest that BQ raises [Ca2+]i by inhibition of Ca2+-ATPases, is a comitogen, and does so at concentrations that could be achieved in vivo in the general urban population.


NPC1L1 (Niemann-Pick C1-like 1) mediates sterol-specific unidirectional transport of non-esterified cholesterol in McArdle-RH7777 hepatoma cells.
Recent evidence suggests that NPC1L1 (Niemann-Pick C1-like 1) is critical for intestinal sterol absorption in mice, yet mechanisms by which NPC1L1 regulates cellular sterol transport are lacking. In the study we used a McArdle-RH7777 rat hepatoma cell line stably expressing NPC1L1 to examine the sterol-specificity and directionality of NPC1L1-mediated sterol transport. As previously described, cholesterol-depletion-driven recycling of NPC1L1 to the cell surface facilitates cellular uptake of non-esterified (free) cholesterol. However, it has no impact on the uptake of esterified cholesterol, indicating free sterol specificity. Interestingly, the endocytic recycling of NPC1L1 was also without effect on beta-sitosterol uptake, indicating that NPC1L1 can differentiate between free sterols of animal and plant origin in hepatoma cells. Furthermore, NPC1L1-driven free cholesterol transport was unidirectional, since cellular cholesterol efflux to apolipoprotein A-I, high-density lipoprotein or serum was unaffected by NPC1L1 expression or localization. Additionally, NPC1L1 facilitates mass non-esterified-cholesterol uptake only when it is located on the cell surface and not when it resides intracellularly. Finally, NPC1L1-dependent cholesterol uptake required adequate intracellular K(+), yet did not rely on intracellular Ca(2+), the cytoskeleton or signalling downstream of protein kinase A, protein kinase C or pertussis-toxin-sensitive G-protein-coupled receptors. Collectively, these findings support the notion that NPC1L1 can selectively recognize non-esterified cholesterol and promote its unidirectional transport into hepatoma cells.


A frequency-based approach to locate common structure for 2D-3D intensity-based registration of setup images in prostate radiotherapy.
In many radiotherapy clinics, geometric uncertainties in the delivery of 3D conformal radiation therapy and intensity modulated radiation therapy of the prostate are reduced by aligning the patient's bony anatomy in the planning 3D CT to corresponding bony anatomy in 2D portal images acquired before every treatment fraction. In this paper, we seek to determine if there is a frequency band within the portal images and the digitally reconstructed radiographs (DRRs) of the planning CT in which bony anatomy predominates over non-bony anatomy such that portal images and DRRs can be suitably filtered to achieve high registration accuracy in an automated 2D-3D single portal intensity-based registration framework. Two similarity measures, mutual information and the Pearson correlation coefficient were tested on carefully collected gold-standard data consisting of a kilovoltage cone-beam CT (CBCT) and megavoltage portal images in the anterior-posterior (AP) view of an anthropomorphic phantom acquired under clinical conditions at known poses, and on patient data. It was found that filtering the portal images and DRRs during the registration considerably improved registration performance. Without filtering, the registration did not always converge while with filtering it always converged to an accurate solution. For the pose-determination experiments conducted on the anthropomorphic phantom with the correlation coefficient, the mean (and standard deviation) of the absolute errors in recovering each of the six transformation parameters were Theta(x):0.18(0.19) degrees, Theta(y):0.04(0.04) degrees, Theta(z):0.04(0.02) degrees, t(x):0.14(0.15) mm, t(y):0.09(0.05) mm, and t(z):0.49(0.40) mm. The mutual information-based registration with filtered images also resulted in similarly small errors. For the patient data, visual inspection of the superimposed registered images showed that they were correctly aligned in all instances. The results presented in this paper suggest that robust and accurate registration can be achieved with intensity-based methods by focusing on rigid bony structures in the images while diminishing the influence of artifacts with similar frequencies as soft tissue.


Viewing time and facial trustworthiness perception: Giving it a second thought may not work for older adults.
Older adults tend to rate unfamiliar faces higher on trustworthiness than do their younger counterparts. Although the saying goes "look before you leap", it is still unknown whether such a strategy could also apply to facial trustworthiness perception, and our understanding of the time course in facial trustworthiness perception also remains unclear. Here, we have argued that a cognitive controlled process suggested by "socioemotional selectivity theory" could potentially lead to such biased trustworthiness perception. Two experiments were conducted to test the association between viewing time and trustworthiness perception. The first study used hierarchical linear modeling in a sample of younger (N = 30, Mage = 20.53, SD = 1.61, 50% female) and older (N = 30, Mage = 63.27, SD = 3.14, 43% female) adults, and found that viewing time and trustworthiness evaluation were positively associated. Using the same stimuli, our second study further manipulated viewing time by two levels (500 ms vs. 3000 ms) and compared younger (N = 28, Mage = 23.93, SD = 2.68, 50% female) and older (N = 30, Mage = 64.47, SD = 4.32, 50% female) adults' facial trustworthiness evaluation. As expected, a significant three-way interaction revealed that viewing time only impacted older adults' facial trustworthiness evaluation, and only when given shorter viewing time did older adults show similar facial trustworthiness ratings as younger adults. The present study is the first to directly investigate the relationship between older adults' viewing time on unfamiliar faces and their perception of facial trustworthiness. Findings suggested that a second thought in facial perception may not benefit older adults' trustworthiness evaluation.


Magnetic resonance imaging evaluation of chronic changes in latissimus dorsi cardiomyoplasty.
Long-term clinical and hemodynamic benefits of dynamic cardiomyoplasty (DC) have been reported. However, no information is available about long-term morphological changes in the wrapped latissimus dorsi (LD) muscle in humans. The LD muscle flap was evaluated by magnetic resonance imaging (MRI) in 13 patients submitted to dynamic cardiomyoplasty for treatment of severe dilated cardiomyopathy. Eight patients were studied 15 days after the surgery (group 1) and 5 patients were studied from 24 to 52 months after the surgical procedure at the time of the cardiomyostimulator replacement (group 2). In the interim, LD was stimulated with burst of 6 pulses (duration, 185 milliseconds; burst frequency, 30 Hz) synchronized to every cardiac contraction, with a maximum of 100 LD contractions per minute. Images were acquired on a GE Sigma 1.5-T system (echo time, 25 milliseconds; repetition time, R-R x 2; slice thickness, 8 mm). The thickness of the LD decreased from 19.6 +/- 7.3 mm for group 1 to 7.6 +/- 0.8 mm for group 2 (P < .01). In addition, the signal intensity of the LD was compared with that of thoracic skeletal muscle and was found to be significantly increased in group 2 (2.19 +/- 0.42) compared with group 1 (1.04 +/- 0.07, P < .001). The signal intensity for group 2 on the T1-weighted images was similar to that of subcutaneous fat in those images. Morphological changes in the wrapped LD muscle consistent with fatty degeneration occur after DC and can be detected by MRI. Further studies will be necessary to demonstrate the clinical significance of such LD muscle flap changes.


Prevalence, Reasons, and Perceived Effects of Khat Chewing Among Students of a College in Gondar Town, Northwestern Ethiopia: A Cross-Sectional Study.
The estimate of the number of people chewing Khat globally ranges from 5 to 10 million people. Its use may result in a variety of effects due to the different compounds in it with effects on the gastro-intestinal system and nervous system being the principal ones. To assess the prevalence, factors, and effects of Khat chewing among students of a college in Gondar town, northwestern Ethiopia. An institution-based cross-sectional study was conducted from 15(th) to 20(th) of April 2009 on a total sample of 424 students who were selected using stratified random sampling technique. Data were collected by three of the principal investigators using a structured pretested data collection instrument and analyzed by Epi Info version 3.5.2. The lifetime and current prevalence of Khat chewing among the respondents were 42% (168/400) and 32.5% (130/400), respectively. Sex (P < 0.01), religion (P < 0.001), and income (P < 0.01) showed statistically significant variation in Khat chewing. The commonest frequency of Khat chewing was once a day 33.1% (43/130) while alcohol (40.8% [53/130]) and cigarette (40.0% [52/130]) were the mostly used substances with Khat. More than half of the chewers (53.85% [70/130]) reported spending 1-4 h for one Khat chewing ceremony. Financially majority of the chewers reported spending up to 10 Ethiopian Birr (ETB) (1.13 United States Dollar) on Khat ( 54.6% [71/130]) and other substances (64.6% [84/130]). Nearly two-thirds (62.3% [81/130]) of the chewers mentioned seeking concentration during study as their main reason for chewing. Among chewers, 83.1% (108/130) reported they faced problem associated to sleep disturbance, 82.3% (107/130) loss of appetite, and 80.8% (105/130) constipation. The prevalence of Khat chewing was fairly high among the students and the majority among them used other substances together with Khat. Spending of a significant amount of money and facing health problems were reported to be consequences of the habit. The college should take steps to make students aware of the ills of Khat chewing and associated habits.


MRC-5 cells, a model for Junín virus persistent infection.
Persistent infection of MRC-5 cells was established following inoculation with attenuated Junín virus (JV). In the acute phase of the infection both the pathogenic XJ and the attenuated XJ0 and XJC13 strains showed severe c.p.e. and free viral titres reached 10(5) p.f.u./ml. Recovery and establishment of persistently infected MRC-5 sublines (MRC-5PI) proved a very common event and seemed to be independent of viral strain, m.o.i. employed or virus passage history. These MRC-5PI sublines released virus throughout their life span and infectious centre assays performed at different passage levels with two sublines showed that 5 to 9% of the cells were producing virus. Heterotypic but not heterologous resistance to superinfection developed, as observed in persistent JV-heteroploid cell systems. Analysis of released JV showed that attenuation had not been markedly altered, but alteration in plaque morphology under methyl cellulose, appearance of temperature-sensitive mutants and alterations in mouse pathology imply that some properties of JV have been altered. Results presented here stress once again the ability of JV to establish persistent infections. The source and diploid characteristics of MRC-5 cells make them a satisfactory model for JV persistence studies.


Clinical significance of early vs late hypotensive blood pressure response to treadmill exercise.
It is generally believed that exercise-induced hypotension is the result of severe left-main or triple-vessel disease. Since this is not invariably so, and since most studies were performed in male populations, this study was done to determine the frequency of, and the significance of, exercise-induced hypotension in a more general population. The treadmill exercise tests of 4850 consecutive patients performed at a university medical center over a period of 7.5 years were reviewed. To identify patients for further analysis, a hypotensive blood pressure response was initially defined (1) as a progressive fall in systolic blood pressure, (2) as a failure of the systolic blood pressure to rise more than 5 mm Hg during exercise, or (3) as an initial rise followed by a fall below the resting standing systolic blood pressure. The incidence of exercise-induced hypotension so defined was less than 2%. Exercise-induced hypotension occurred in two patterns. An early hypotension response was defined as a fall in systolic blood pressure of more than 10 mm Hg, associated with symptoms or ST-segment depression, during the first 5 minutes of exercise or as a progressive fall in systolic blood pressure of at least 20 mm Hg. The majority of patients (nine of 10) with an early hypotensive response had severe coronary artery disease. The late hypotension pattern was characterized by an initial rise, followed by a fall in the systolic blood pressure with continued exercise. Only half of the patients with this pattern had significant coronary artery disease, and half of the patients had other causes for exercise-induced hypotension. A late hypotensive response was six times more frequent than an early hypotensive response. This study identified two patterns of exercise-induced hypotension. Early, almost always due to severe coronary artery disease, and late, six times more common than early in which only half were due to coronary artery disease. Causes of a late hypotensive response to exercise that were not due to severe coronary artery disease included valvular heart disease, orthostatic hypotension, cardiomyopathy, idiopathic causes, and drugs. Drugs that contributed to a late exercise-induced hypotension response were diuretics, vasodilators, and negative inotropic agents.


Engagement of β-arrestin by transactivated insulin-like growth factor receptor is needed for V2 vasopressin receptor-stimulated ERK1/2 activation.
G protein-coupled receptors (GPCRs) have been shown to activate the mitogen-activated protein kinases, ERK1/2, through both G protein-dependent and -independent mechanisms. Here, we describe a G protein-independent mechanism that unravels an unanticipated role for β-arrestins. Stimulation of the V2 vasopressin receptor (V2R) in cultured cells or in vivo in rat kidney medullar collecting ducts led to the activation of ERK1/2 through the metalloproteinase-mediated shedding of a factor activating the insulin-like growth factor receptor (IGFR). This process was found to be both Src- and β-arrestin-dependent. Whereas Src was found to act upstream of the metalloproteinase activation and be required for the release of the IGFR-activating factor, β-arrestins were found to act downstream of the IGFR transactivation. Unexpectedly, the engagement of β-arrestins by the IGFR but not by the V2R was needed to promote the vasopressin-stimulated ERK1/2 activation, indicating that a pool of β-arrestins distinct from those β-arrestins recruited to the V2R acts downstream of the receptor tyrosine kinase to activate ERK1/2. Such a dual site of action for β-arrestins helps explain the pleiotropic actions of this scaffolding protein. Given the role that V2R-stimulated ERK1/2 plays in kidney cell proliferation, this transactivation mechanism may have important implications for renal pathophysiology. Still, the role of β-arrestins downstream of a transactivation event is not limited to the V2R, because we observed a similar involvement for an unrelated GPCR (the platelet-activating factor receptor), indicating that it may be a general mechanism shared among GPCRs.


Genotype-phenotype correlation of paroxysmal nonkinesigenic dyskinesia.
Paroxysmal nonkinesigenic dyskinesia (PNKD) is a rare disorder characterized by episodic hyperkinetic movement attacks. We have recently identified mutations in the MR-1 gene causing familial PNKD. We reviewed the clinical features of 14 kindreds with familial dyskinesia that was not clearly induced by movement or during sleep. Of these 14 kindreds, 8 had MR-1 mutations and 6 did not. Patients with PNKD with MR-1 mutations had their attack onset in youth (infancy and early childhood). Typical attacks consisted of a mixture of chorea and dystonia in the limbs, face, and trunk, and typical attack duration lasted from 10 minutes to 1 hour. Caffeine, alcohol, and emotional stress were prominent precipitants. Attacks had a favorable response to benzodiazepines, such as clonazepam and diazepam. Attacks in families without MR-1 mutations were more variable in their age at onset, precipitants, clinical features, and response to medications. Several were induced by persistent exercise. Paroxysmal nonkinesigenic dyskinesia (PNKD) should be strictly defined based on age at onset and ability to precipitate attacks with caffeine and alcohol. Patients with this clinical presentation (which is similar to the phenotype initially reported by Mount and Reback) are likely to harbor myofibrillogenesis regulator 1 (MR-1) gene mutations. Other "PNKD-like" families exist, but atypical features suggests that these subjects are clinically distinct from PNKD and do not have MR-1 mutations. Some may represent paroxysmal exertional dyskinesia.


Learners' decisions for attending Pediatric Grand Rounds: a qualitative and quantitative study.
Although grand rounds plays a major educational role at academic medical centers, there has been little investigation into the factors influencing the learners' decision to attend. Greater awareness of attendees' expectations may allow grand rounds planners to better accommodate the learners' perspective, potentially making continuing education activities more attractive and inviting. We used both qualitative (part A) and quantitative (part B) techniques to investigate the motivators and barriers to grand rounds attendance. Part A investigated contextual factors influencing attendance as expressed through attendee interviews. Transcripts of the interviews were analyzed using grounded theory techniques. We created a concept map linking key factors and their relationships. In part B we quantified the motivators and barriers identified during the initial interviews through a survey of the grand rounds audience. Sixteen persons voluntarily took part in the qualitative study (part A) by participating in one of seven group interview sessions. Of the 14 themes that emerged from these sessions, the most frequent factors motivating attendance involved competent practice and the need to know. All sessions discussed intellectual stimulation, social interaction, time constraints and convenience, licensure, content and format, and absence of cost for attending sessions. The 59 respondents to the survey (part B) identified clinically-useful topics (85%), continuing education credit (46%), cutting-edge research (27%), networking (22%), and refreshments (8%) as motivators and non-relevant topics (44%) and too busy to attend (56%) as barriers. Greater understanding of the consumers' perspective can allow planners to tailor the style, content, and logistics to make grand rounds more attractive and inviting.


Doorway states in the gamma decay-out of the yrast superdeformed band in 59Cu.
The decay-out process of the yrast superdeformed band in 59Cu has been investigated. The firm determination of spin, parity, excitation energy, and configuration of the states involved in this process constitutes a unique situation for a detailed understanding of the decay-out mechanism. A theoretical model is introduced that includes a residual interaction and tunneling matrix element between bands, calculated in the configuration-dependent cranked Nilsson-Strutinsky model. This interaction causes the decay to occur via a small number of observed doorway states.


Occupational stress and coping in a sample of student nurses.
The study investigated the extent and nature of work stress and burnout in a sample of 81 student nurses in a training hospital in the Natal Midlands. The Maslach Burnout Inventory (MBI), Sources of Stress Questionnaire (SSQ), Student Life Events Checklist (SLEC), and two coping scales, the Ways of Coping Checklist (WCC) and the Billings and Moos Coping Scale (BMCS) were used to measure the variable of burnout, stress, life events and ways of coping. Findings point to the fact that there were significantly higher levels of burnout as measured on the MBI sub-scales of Emotional Exhaustion and Depersonalisation in the sample, in comparison to other helping professionals in South Africa and the USA. These results are discussed in relation to the influence of gender, race and age of the sample. A small sub-sample of high burnout nurses was identified that differed from the rest of the sample in respect to coping styles. Sources of stress in the sample indicated issues relevant to the developmental stage of the sample that have implications for training and intervention. Style of coping and source of stress relating to home-work interface were found to be related to the experience of burnout. Implications of the findings are discussed in relation to training, intervention and further research.


Conophylline suppresses pancreatic cancer desmoplasia and cancer-promoting cytokines produced by cancer-associated fibroblasts.
Despite recent advances in cancer treatment, pancreatic cancer is a highly malignant tumor type with a dismal prognosis and it is characterized by dense desmoplasia in the cancer tissue. Cancer-associated fibroblasts (CAF) are responsible for this fibrotic stroma and promote cancer progression. We previously reported that a novel natural compound conophylline (CnP) extracted from the leaves of a tropical plant reduced liver and pancreatic fibrosis by suppression of stellate cells. However, there have been no studies to investigate the effects of CnP on CAF, which is the aim of this work. Here, we showed that CAF stimulated indicators of pancreatic cancer malignancy, such as proliferation, invasiveness, and chemoresistance. We also showed that CnP suppressed CAF activity and proliferation, and inhibited the stimulating effects of CAF on pancreatic cancer cells. Moreover, CnP strongly decreased the various cytokines involved in cancer progression, such as interleukin (IL)-6, IL-8, C-C motif chemokine ligand 2 (CCL2), and C-X-C motif chemokine ligand 12 (CXCL12), secreted by CAF. In vivo, CAF promoted tumor proliferation and desmoplastic formation in a mouse xenograft model, CnP reduced desmoplasia of tumors composed of pancreatic cancer cells + CAF, and combination therapy of CnP with gemcitabine remarkably inhibited tumor proliferation. Our findings suggest that CnP is a promising therapeutic strategy of combination therapy with anticancer drugs to overcome refractory pancreatic cancers.


New complex of post-activated neocarzinostatin chromophore with DNA: bulge DNA binding from the minor groove.
Neocarzinostatin (NCS-chrom), a natural enediyne antitumor antibiotic, undergoes either thiol-dependent or thiol-independent activation, resulting in distinctly different DNA cleavage patterns. Structures of two different post-activated NCS-chrom complexes with DNA have been reported, revealing strikingly different binding modes that can be directly related to the specificity of DNA chain cleavage caused by NCS-chrom. The third structure described herein is based on recent studies demonstrating that glutathione (GSH) activated NCS-chrom efficiently cleaves DNA at specific single-base sites in sequences containing a putative single-base bulge. In this structure, the GSH post-activated NCS-chrom (NCSi-glu) binds to a decamer DNA, d(GCCAGAGAGC), from the minor groove. This binding triggers a conformational switch in DNA from a loose duplex in the free form to a single-strand, tightly folded hairpin containing a bulge adenosine embedded between a three base pair stem. The naphthoate aromatic moiety of NCSi-glu intercalates into a GG step flanked by the bulge site, and its substituent groups, the 2-N-methylfucosamine carbohydrate ring and the tetrahydroindacene, form a complementary minor groove binding surface, mostly interacting with the GCC strand in the duplex stem of DNA. The bulge site is stabilized by the interactions involving NCSi-glu naphthoate and GSH tripeptide. The positioning of NCSi-glu is such that only single-chain cleavage via hydrogen abstraction at the 5'-position of the third base C (which is opposite to the putative bulge base) in GCC is possible, explaining the observed single-base cleavage specificity. The reported structure of the NCSi-glu-bulge DNA complex reveals a third binding mode of the antibiotic and represents a new family of minor groove bulge DNA recognition structures. We predict analogue structures of NCSi-R (R = glu or other substituent groups) may be versatile probes for detecting the existence of various structures of nucleic acids. The NMR structure of this complex, in combination with the previously reported NCSi-gb-bulge DNA complex, offers models for specific recognition of DNA bulges of various sizes through binding to either the minor or the major groove and for single-chain cleavage of bulge DNA sequences.


Using the Star Excursion Balance Test to assess dynamic postural-control deficits and outcomes in lower extremity injury: a literature and systematic review.
A dynamic postural-control task that has gained notoriety in the clinical and research settings is the Star Excursion Balance Test (SEBT). Researchers have suggested that, with appropriate instruction and practice by the individual and normalization of the reaching distances, the SEBT can be used to provide objective measures to differentiate deficits and improvements in dynamic postural-control related to lower extremity injury and induced fatigue, and it has the potential to predict lower extremity injury. However, no one has reviewed this body of literature to determine the usefulness of the SEBT in clinical applications. To provide a narrative review of the SEBT and its implementation and the known contributions to task performance and to systematically review the associated literature to address the SEBT's usefulness as a clinical tool for the quantification of dynamic postural-control deficits from lower extremity impairment. Databases used to locate peer-reviewed articles published from 1980 and 2010 included Derwent Innovations Index, BIOSIS Previews, Journal Citation Reports, and MEDLINE. The criteria for article selection were (1) The study was original research. (2) The study was written in English. (3) The SEBT was used as a measurement tool. Specific data extracted from the articles included the ability of the SEBT to differentiate pathologic conditions of the lower extremity, the effects of external influences and interventions, and outcomes from exercise intervention and to predict lower extremity injury. More than a decade of research findings has established a comprehensive portfolio of validity for the SEBT, and it should be considered a highly representative, noninstrumented dynamic balance test for physically active individuals. The SEBT has been shown to be a reliable measure and has validity as a dynamic test to predict risk of lower extremity injury, to identify dynamic balance deficits in patients with a variety of lower extremity conditions, and to be responsive to training programs in both healthy people and people with injuries to the lower extremity. Clinicians and researchers should be confident in employing the SEBT as a lower extremity functional test.


Reliability and validity of the multi-point method and the 2-point method's variations of estimating the one-repetition maximum for deadlift and back squat exercises.
This study aimed at examining the concurrent validity and reliability of the multi-point method and the two-point method's variations for estimating the one-repetition maximum (1RM) in the deadlift and squat exercises and to determine the accuracy of which optimal two loads can be used for the two-point method protocol. Thirteen resistance-trained men performed six sessions that consisted of two incremental loading tests (multi-point method: 20-40-60-80-90% and two-point method variations: 40-60%, 40-80%, 40-90%,60-80%, 60-90%) followed by 1RM tests. Both the multi-point method and the two-point method load variations showed reliable results for 1RM estimation (CV < 10%) squat and deadlift exercises. Session-session reliability was found to be low in deadlift (ICC: 0.171-0.335) and squat exercises (ICC: 0.235-0.479) of 40-60% and 60-80% in two-point methods. Deadlift (ICC: 0.815-0.996) and squat (ICC: 0.817-0.988) had high session-to-session reliability in all other methods. Regarding the validity of deadlift exercise, the multipoint method (R2 = 0.864) and two variations of the two-point method (R2 = 0.816 for 40-80%, R2 = 0.732 for 60-80%) showed very large correlations, whereas other two variations of the two-point method (R2 = 0.945 for 40-90%, R2 = 0.914 for 60-90%) showed almost perfect correlations with the actual 1RM. Regarding the validity of squat exercise, the multi-point method (R2 = 0.773) and two variations of the two-point method (R2 = 0.0847 for 60-80%, R2 = 0.705 for 40-90%) showed very large correlations, whereas 40-60% variation showed almost perfect correlation (R2 = 0.962) with the actual 1RM. In conclusion, whereas both the multi-point method and the two-point method load variations showed reliable results, the multiple-point method and most of the two-point methods' load variations examined in this research provided an accurate (from large-moderate to perfect) estimate of the 1RM. Therefore, we recommend using the multi-point method and especially the two-point methods variations including higher relative loads to estimate 1RM.


The impact of insulin type on severe hypoglycaemia events requiring inpatient and emergency department care in patients with type 2 diabetes.
To evaluate the risk from different insulin types on severe hypoglycaemia (SHG) events requiring inpatient (IP) or emergency department (ED) care in patients with type 2 diabetes. Type 2 diabetes patients newly started on insulin in a large commercial claims database were evaluated for SHG events. Patients were classified into an insulin group based on their most frequently used insulin type. Multivariable Cox models assessed the association between insulin type and the risk of SHG events. We identified 8626 patients (mean age 53.5 years; 55% female) with type 2 diabetes followed for an average of 4.0 years after insulin initiation. Of these, 161 (1.9%) had a SHG event at an average of 3.1y after insulin initiation. Patients with SHG events were slightly older (56.4 vs. 53.4 years), used a similar number of OADs (1.1 vs. 1.2) but had more co-morbidities compared with those without SHG events. In multivariate Cox models, premixed insulin (HR 2.12; p<0.01), isophane insulin (NPH) (HR 2.02; p<0.01), and rapid acting insulin (HR 2.75; p<0.01) had significantly higher risks of SHG events compared with glargine. No statistically significant difference in SHG events was seen with detemir (HR 1.20; p=0.73). Among patients with type 2 diabetes, the use of newer basal insulin analogues was associated with lower rates of SHG events requiring IP or ED care compared with users of other insulin formulations. Future research should examine the impact of hypoglycaemia events of different severity levels.


Effects of Meteorological Conditions on PM2.5 Concentrations in Nagasaki, Japan.
The fine particulate matter (PM2.5) problem has attracted much scientific and public attention, due to its effects on visibility, human health, and global climate. There are three factors that have important effect on PM2.5 mass concentration: domestic pollutant emission sources, external sources outside of the country, and the meteorological conditions. Nagasaki is a coastal prefecture located at the westernmost part of Japan, which is an ideal location to study pollutants from long range transport and correlation between PM2.5 and meteorological conditions. In this paper, PM2.5 concentration data and meteorological data were obtained during 1 January 2013~31 December 2013. The spatial distribution depicts that the western part of the study area has the most serious PM2.5 pollution. The correlation analysis results between PM2.5 concentration and meteorological data showed that temperature had a negative, and precipitation had a positive, correlation with PM2.5. There was a threshold in the correlations between humidity and wind speed and PM2.5. The correlation was positive or negative depending on the meteorological variable values, if these were lower or higher than the threshold. From the relationship with wind direction, it can be depicted that the west wind might bring the most pollutants to Nagasaki.


Reference Values for Central Airway Dimensions on CT Images of Children and Adolescents.
The purpose of this study was to acquire normative data on central airway dimensions on chest CT scans in the pediatric population. Chest CT findings reported as normal by a radiologist were collected retrospectively at 10 international centers. An experienced and independent thoracic radiologist reevaluated all CT scans for image quality and for normal findings. Semiautomated image analysis was performed to measure dimensions of the trachea and right and left main bronchi at inspiration. Intrathoracic tracheal length was measured from carina to thorax inlet. Cross-sectional area and short and long axes were measured perpendicular to the longitudinal airway axis starting from the carina every centimeter upward for the trachea and every 0.5 cm downward for the main bronchi. The effects on airway diameters of age, sex, intrathoracic tracheal length, and distance from the carina were investigated by use of mixed-effects models analysis. Among 1160 CT scans collected, 388 were evaluated as normal by the independent radiologist with sufficient image quality and adequate inspiratory volume level. Central airways were successfully semiautomatically analyzed in 294 of 388 CT studies. Age, sex, intrathoracic tracheal length, and distance from carina were all significant predictors in the models for tracheal and right and left main bronchial diameters (p < 0.001). The central airway dimensions increased with age up to 20 years, and dimensions were larger in male than in female adolescents. Normative data were determined for the central airways of children and adolescents. Central airway dimensions depended on distance from the carina and on intrathoracic tracheal length.


A1 and A2 erythrocytes can be distinguished by reagents that do not detect structural differences between the two cell types.
The differential reactivity of four mouse monoclonal antibodies (AbCB, AbHT29-36, AbM2, and AbS12) and Dolichos biflorus lectin with A1 and A2 erythrocytes was analyzed. Only AbS12 and D. biflorus lectin were able to preferentially agglutinate A1 erythrocytes. AbS12 is known to react only with short chain, unbranched structures (such as Aa-2 and Ab-2 glycolipids) and not with longer chains or with type 3 and type 4 structures. D. biflorus was shown to have a similar specificity by lectin staining of glycolipids separated by thin-layer chromatography. Analysis of the binding of radiolabeled AbCB and AbS12 to A1 and A2 erythrocytes by Scatchard analysis showed that, whereas the former antibody recognizes high-affinity sites on both A1 and A2 cells, AbS12 reacts with high-affinity sites only on A1 cells. Because A1 and A2 erythrocytes have a similar complement of short chain type 2 glycolipids, although in different amounts, it is suggested that AbS12 and D. biflorus lectin differentiate between the two cell types on the basis of quantitative, nonstructural features. This is in contrast to AbTH1, which reacts with a repetitive A epitope (type 3 A chain) and distinguishes between A1 and A2 cells based on the preferential expression of type 3 A chains in A1 erythrocytes. Thus, two views of A1/A2, i.e., qualitative vs quantitative are correct, depending on the properties of the reagent being used to distinguish between the two cell types.


'Just like the fact that I'm Syrian like you are Scottish': ascribing interviewer identities as a resource in cross-cultural interaction.
Interviews are used as a research tool in numerous studies of identity. They have also been studied as social interactions in themselves. From this perspective, analysts have looked at the ways interviews generate identities, and how both parties work to fulfill the associated roles and expectations (e.g., the interviewer's neutrality). Interviewers thus become a more 'visible' participant, and several studies examine the influence of their multiple category memberships on the interaction. This paper aims to extend these insights to the insider/outsider dilemma in interviews, by looking at how participants make relevant and use interviewer identities as a resource. The data comes from a corpus of interviews with Syrian people on identity conducted by a British interviewer. The analysis shows how the interviewer's identity as a sojourner and stranger was used to warrant positive assessments of national character; her national identity was invoked to normalize national feelings; her identity as Christian used to characterize religious identity as inherent; and her identity as Westerner was used to legitimate complaints. I discuss implications for interviews on identity and identity within interviews, and argue that this approach is a thoroughly empirical way to address the shifting, practical ascription of insider-outsider status to the interviewer.


[Long-term mechanical ventilation survey].
Although home mechanical ventilation is common in western countries, it is still extremely rare in Taiwan. The objectives of this study are (1) to review retrospectively the types of disorders and outcomes of these patients requiring prolonged mechanical ventilation (MV), and (2) to survey the home status of twelve patients on home mechanical ventilation. From January 1990 to April 1992, one hundred fifteen patients receiving prolonged MV ( > or = 30 days & > or = 8-10 hrs/day) were studied in the medical intensive care unit at Chang Gung Memorial Hospital. Eighty-two patients (71.3%) for whom long-term MV were thought to be beneficial were divided into three groups by disorder: (1)primary pulmonary disorder (N = 55, 67.07%) (2) neuromuscular disorder (N = 18, 21.95%), (3) brain stem disorder (N = 9, 10.98%). The outcomes of these eighty-two patients were as follows: expired, 37 (45.12%), completely weaned, 21 (25.61%), unweanable 24, including 12 (14.63%) hospitalized and 12 (14.63%) discharged home. The three outcomes of prolonged MV were significantly associated with the three types of disorder (P = 0.0035). Among these three disorders, duration of MV were longest in the neuromuscular group, whatever the outcome. There was significant difference in the duration of MV for the different outcomes of group I & group III. The mean duration at home for the twelve home MV patients (5 male and 7 female, mean age +/- SD = 46.08 +/- 20.83 years old) was 9.93 +/- 8.11 month, with the longest being 21.5 months. Four expired within two months of discharge to home, two patients had to be readmitted once (one C2-C5 spinal cord injury patient for pneumonia, and the other for brain stem deterioration). When comparing hospital stay, four patients improved in the maximal free time (the mean improvement, 5.75 +/- 3.11 hours/day), seven remained unchanged, while one regressed.(ABSTRACT TRUNCATED AT 250 WORDS)


Cyclooctatetraene-conjugated cyanine mitochondrial probes minimize phototoxicity in fluorescence and nanoscopic imaging.
Modern fluorescence-imaging methods promise to unveil organelle dynamics in live cells. Phototoxicity, however, has become a prevailing issue when boosted illumination applies. Mitochondria are representative organelles whose research heavily relies on optical imaging, yet these membranous hubs of bioenergy are exceptionally vulnerable to photodamage. We report that cyclooctatetraene-conjugated cyanine dyes (PK Mito dyes), are ideal mitochondrial probes with remarkably low photodynamic damage for general use in fluorescence cytometry. In contrast, the nitrobenzene conjugate of Cy3 exhibits enhanced photostability but unaffected phototoxicity compared to parental Cy3. PK Mito Red, in conjunction with Hessian-structural illumination microscopy, enables 2000-frame time-lapse imaging with clearly resolvable crista structures, revealing rich mitochondrial dynamics. In a rigorous stem cell sorting and transplantation assay, PK Mito Red maximally retains the stemness of planarian neoblasts, exhibiting excellent multifaceted biocompatibility. Resonating with the ongoing theme of reducing photodamage using optical approaches, this work advocates the evaluation and minimization of phototoxicity when developing imaging probes.


De-icing: recovery of diffraction intensities in the presence of ice rings.
Macromolecular structures are routinely determined at cryotemperatures using samples flash-cooled in the presence of cryoprotectants. However, sometimes the best diffraction is obtained under conditions where ice formation is not completely ablated, with the result that characteristic ice rings are superimposed on the macromolecular diffraction. In data processing, the reflections that are most affected by the ice rings are usually excluded. Here, an alternative approach of subtracting the ice diffraction is tested. High completeness can be retained with little adverse effect upon the quality of the integrated data. This offers an alternate strategy when high levels of cryoprotectant lead to loss of crystal quality.


The use of flow cytometry in assessing malignancy in bone and soft tissue tumors.
Since 1982, the orthopaedic research laboratories at the authors' hospital has done flow cytometric and more recently cytofluorometric deoxyribonucleic ploidic analyses of samples of bone and soft tissue tumors. The current authors attempt to define the value of such studies in distinguishing benign from malignant tumors, in conforming to stage of the tumors, and in helping to predict metastasis and death. The series consists of 1134 patients in whom the disease was verified and the survival data were available as a result of a questionnaire study. Statistically, the ploidic analyses were of remarkable value in defining malignancy and in correlating with the stage of the lesion. They were of less value in predicting survival, particularly for patients with osteosarcoma and chondrosarcoma, but seemed to predict survival effectively for patients with soft tissue sarcomas.


[En epidemiologic approach for the analysis of the demand and supply of psycho-psychiatric care in general medicine and in specialized services].
In this paper we present data collected in the area of Verona concerning indicators of demand and of supply of care in two settings, the general practice and the South-Verona Community Psychiatric Service. These data, obtained from the epidemiological studies conducted by our research group during the past ten years, emphasize similarities and differences existing in the two levels of care. In general practice the demand for care for psychological disorders is difficult to identify, because of the frequent somatic expression of these disorders; moreover at the level of psychiatric services the demand is frequently aspecific. As far as the supply of care is concerned, in both settings drug prescription is the most frequent treatment, however other interventions are also offered. For example, in general practice counselling and referral to the psychiatrist are often provided; in psychiatric services monitored during our studies a large range of treatments were supplied, often in an integrated way.


Population genomics of transposable elements in Drosophila melanogaster.
Transposable elements (TEs) are the primary contributors to the genome bulk in many organisms and are major players in genome evolution. A clear and thorough understanding of the population dynamics of TEs is therefore essential for full comprehension of the eukaryotic genome evolution and function. Although TEs in Drosophila melanogaster have received much attention, population dynamics of most TE families in this species remains entirely unexplored. It is not clear whether the same population processes can account for the population behaviors of all TEs in Drosophila or whether, as has been suggested previously, different orders behave according to very different rules. In this work, we analyzed population frequencies for a large number of individual TEs (755 TEs) in five North American and one sub-Saharan African D. melanogaster populations (75 strains in total). These TEs have been annotated in the reference D. melanogaster euchromatic genome and have been sampled from all three major orders (non-LTR, LTR, and TIR) and from all families with more than 20 TE copies (55 families in total). We find strong evidence that TEs in Drosophila across all orders and families are subject to purifying selection at the level of ectopic recombination. We showed that strength of this selection varies predictably with recombination rate, length of individual TEs, and copy number and length of other TEs in the same family. Importantly, these rules do not appear to vary across orders. Finally, we built a statistical model that considered only individual TE-level (such as the TE length) and family-level properties (such as the copy number) and were able to explain more than 40% of the variation in TE frequencies in D. melanogaster.


An FGFR3 Autocrine Loop Sustains Acquired Resistance to Trastuzumab in Gastric Cancer Patients.
The majority of gastric cancer patients who achieve an initial response to trastuzumab-based regimens develop resistance within 1 year of treatment. This study was aimed at identifying the molecular mechanisms responsible for resistance. A HER2+-trastuzumab sensitive NCI-N87 gastric cancer orthotopic nude mouse model was treated with trastuzumab until resistance emerged. Differentially expressed transcripts between trastuzumab-resistant and sensitive gastric cancer cell lines were annotated for functional interrelatedness by Ingenuity Pathway Analysis software. Immunohistochemical analyses were performed in pretreatment versus posttreatment biopsies from gastric cancer patients receiving trastuzumab-based treatments. All statistical tests were two-sided. Four NCI-N87 trastuzumab-resistant (N87-TR) cell lines were established. Microarray analysis showed HER2 downregulation, induction of epithelial-to-mesenchymal transition, and indicated fibroblast growth factor receptor 3 (FGFR3) as one of the top upregulated genes in N87-TR cell lines. In vitro, N87-TR cell lines demonstrated a higher sensitivity than did trastuzumab-sensitive parental cells to the FGFR3 inhibitor dovitinib, which reduced expression of pAKT, ZEB1, and cell migration. Oral dovitinib significantly (P = 0.0006) reduced tumor burden and prolonged mice survival duration in N87-TR mouse models. A higher expression of FGFR3, phosphorylated AKT, and ZEB1 were observed in biopsies from patients progressing under trastuzumab-based therapies if compared with matched pretreatment biopsies. This study identified the FGFR3/AKT axis as an escape pathway responsible for trastuzumab resistance in gastric cancer, thus indicating the inhibition of FGFR3 as a potential strategy to modulate this resistance. Clin Cancer Res; 22(24); 6164-75. ©2016 AACR.


Post-translation control of Nramp metal transport in yeast. Role of metal ions and the BSD2 gene.
The Saccharomyces cerevisiae SMF1 gene encodes a member of the well conserved family of Nramp metal transport proteins. Previously, we determined that heavy metal uptake by Smf1p was down-regulated by the product of the S. cerevisiae BSD2 gene. We now demonstrate that this regulation occurs at the level of protein stability. In wild type strains, the bulk of Smf1p is normally directed to the vacuole and is rapidly degraded by vacuolar proteases in a PEP4-dependent manner. In bsd2Delta mutants, Smf1p fails to enter the vacuole, and the Nramp protein is stabilized. Metal ions themselves play an important role in the post-translational regulation of Smf1p. The depletion of heavy metals from the growth medium effects stabilization of Smf1p and additionally results in accumulation of this transporter at the cell surface. Supplementation of manganese alone is sufficient to trigger rapid degradation of Smf1p in a Bsd2p-dependent manner. Together the action of Bsd2p and metal ions provide a rapid and effective means for controlling Nramp metal transport in response to environmental changes.


Procoagulant alveolar microparticles in the lungs of patients with acute respiratory distress syndrome.
Coagulation and fibrinolysis abnormalities are observed in acute lung injury (ALI) in both human disease and animal models and may contribute to ongoing inflammation in the lung. Tissue factor (TF), the main initiator of the coagulation cascade, is upregulated in the lungs of patients with ALI/acute respiratory distress syndrome (ARDS) and likely contributes to fibrin deposition in the air space. The mechanisms that govern TF upregulation and activation in the lung are not well understood. In the vascular space, TF-bearing microparticles (MPs) are central to clot formation and propagation. We hypothesized that TF-bearing MPs in the lungs of patients with ARDS contribute to the procoagulant phenotype in the air space during acute injury and that the alveolar epithelium is one potential source of TF MPs. We studied pulmonary edema fluid collected from patients with ARDS compared with a control group of patients with hydrostatic pulmonary edema. Patients with ARDS have higher concentrations of MPs in the lung compared with patients with hydrostatic edema (25.5 IQR 21.3-46.9 vs. 7.8 IQR 2.3-27.5 micromol/l, P = 0.009 by Mann-Whitney U-test). These MPs are enriched for TF, have procoagulant activity, and likely originate from the alveolar epithelium [as measured by elevated levels of RAGE (receptor for advanced glycation end products) in ARDS MPs compared with hydrostatic MPs]. Furthermore, alveolar epithelial cells in culture release procoagulant TF MPs in response to a proinflammatory stimulus. These findings suggest that alveolar epithelial-derived MPs are one potential source of TF procoagulant activity in the air space in ARDS and that epithelial MP formation and release may represent a unique therapeutic target in ARDS.


A comprehensive integrated drug similarity resource for in-silico drug repositioning and beyond.
Drug similarity studies are driven by the hypothesis that similar drugs should display similar therapeutic actions and thus can potentially treat a similar constellation of diseases. Drug-drug similarity has been derived by variety of direct and indirect sources of evidence and frequently shown high predictive power in discovering validated repositioning candidates as well as other in-silico drug development applications. Yet, existing resources either have limited coverage or rely on an individual source of evidence, overlooking the wealth and diversity of drug-related data sources. Hence, there has been an unmet need for a comprehensive resource integrating diverse drug-related information to derive multi-evidenced drug-drug similarities. We addressed this resource gap by compiling heterogenous information for an exhaustive set of small-molecule drugs (total of 10 367 in the current version) and systematically integrated multiple sources of evidence to derive a multi-modal drug-drug similarity network. The resulting database, 'DrugSimDB' currently includes 238 635 drug pairs with significant aggregated similarity, complemented with an interactive user-friendly web interface (http://vafaeelab.com/drugSimDB.html), which not only enables database ease of access, search, filtration and export, but also provides a variety of complementary information on queried drugs and interactions. The integration approach can flexibly incorporate further drug information into the similarity network, providing an easily extendable platform. The database compilation and construction source-code has been well-documented and semi-automated for any-time upgrade to account for new drugs and up-to-date drug information.


Role of cell surface signaling in proteolysis of an alternative sigma factor in Pseudomonas aeruginosa.
Alternative sigma factor proteins enable transcription of specific sets of genes in bacterial cells. Their activities can be controlled by posttranslational mechanisms including inhibition by antisigma proteins and proteolytic degradation. PvdS is an alternative sigma factor that is required for expression of genes involved in synthesis of a siderophore, pyoverdine, by Pseudomonas aeruginosa. In the absence of pyoverdine, the activity of PvdS is inhibited by a membrane-spanning antisigma factor, FpvR. Inhibition is relieved by a cell surface signaling pathway. In this pathway, a combination of pyoverdine and a cell surface receptor protein, FpvA, suppresses the antisigma activity of FpvR, enabling transcription of PvdS-dependent genes. In this research, we investigated proteolytic degradation of PvdS in response to the signaling pathway. Proteolysis of PvdS was observed in strains of P. aeruginosa in which FpvR had anti-sigma factor activity due to the absence of pyoverdine or the FpvA receptor protein or overproduction of FpvR. Suppression of antisigma activity by addition of pyoverdine or through the absence of FpvR prevented detectable proteolysis of PvdS. The amounts of PvdS were less in bacteria in which proteolysis was observed, and reporter gene assays showed that this reduction was not due to decreased expression of PvdS. In wild-type bacteria, there was an average of 730 molecules of PvdS per cell in late exponential growth phase. Our results show that proteolysis and amounts of PvdS are affected by the antisigma factor FpvR and that this activity of FpvR is controlled by the cell surface signaling pathway.


Bonding efficacy of new self-etching, self-adhesive dual-curing resin cements to dental enamel.
This study evaluated the efficacy of the union between two new self-etching self-adhesive resin cements and enamel using the microtensile bond strength test. Buccal enamel of 80 bovine teeth was submitted to finishing and polishing with metallographic paper to a refinement of #600, in order to obtain a 5-mm2 flat area. Blocks (2 x 4 x 4 mm) of laboratory composite resin were cemented to enamel according to different protocols: (1) untreated enamel + RelyX Unicem cement (RX group); (2) untreated enamel + Bifix SE cement (BF group); (3) enamel acid etching and application of resin adhesive Single Bond + RelyX Unicem (RXA group); (4) enamel acid etching and application of resin adhesive Solobond M + Bifix SE (BFA group). After 7 days of storage in distillated water at 37°C, the blocks were sectioned for obtaining microbar specimens with an adhesive area of 1 mm2 (n = 120). Specimens were submitted to the microtensile bond strength test at a crosshead speed of 0.5 mm/min. The results (in MPa) were analyzed statistically by ANOVA and Tukey's test. Enamel pre-treatment with phosphoric acid and resin adhesive (27.9 and 30.3 for RXA and BFA groups) significantly improved (p ≤ 0.05) the adhesion of both cements to enamel compared to the union achieved with as-polished enamel (9.9 and 6.0 for RX and BF). Enamel pre-treatment with acid etching and the application of resin adhesive significantly improved the bond efficacy of both luting agents compared to the union achieved with as-polished enamel.


Parent and Pediatrician Preferences for Type 1 Diabetes Screening in the U.S.
The purpose of this study was to use a discrete-choice experiment methodology to understand the relative importance of the attributes of screening tests for type 1 diabetes among parents and pediatricians in the U.S. Online surveys presented hypothetical screening test profiles from which respondents chose their preferred test profile. Survey attributes were based on likely screening test options and included the mode of administration, where and when the test was conducted, the type of education and monitoring available to lower the risk of diabetic ketoacidosis (DKA), and whether a treatment was available that would delay onset of insulin dependence. Data were analyzed using random-parameters logit models. Parents placed the highest relative importance on monitoring programs that could reduce the risk of DKA to 1%, followed by treatment to delay onset of insulin dependence by 1 or 2 years, and, finally, avoiding a $50 out-of-pocket cost. Pediatricians placed equal importance on monitoring programs that reduced a patient's risk of DKA to 1% and on avoiding a $50 out-of-pocket cost for the screening test, followed by the option of a treatment to delay the onset of insulin dependence. The mode of administration and location and timing of the screening were much less important to parents and pediatricians. Parents and pediatricians preferred screening tests that were accompanied by education and monitoring plans to reduce the risk of DKA, had available treatment to delay type 1 diabetes, and had lower out-of-pocket costs.


A tandem duplication causes the Kn1-O allele of Knotted, a dominant morphological mutant of maize.
Molecular and genetic techniques are used to define Kn1-O, a mutation which interferes with the normal differentiation of vascular tissue in leaves. Sequences associated with a previously cloned allele, Kn1-2F11, were used as hybridization probes in a Southern analysis of Kn1-O. By this analysis, Kn1-O lacks the Ds2 transposable element that causes Kn1-2F11 but instead is associated with a sequence duplication. Sequence and restriction analysis of genomic clones show that the duplication consists of a tandem array of two 17-kb repeats. Analysis of Kn1-O derivatives indicates that the duplication itself conditions the mutant phenotype; a severely knotted line, Kn1-Ox, has gained a repeat unit to form a triplication, whereas normal derivatives have either lost a repeat unit or sustained insertions that disrupt the tandem duplication. These insertions map near the central junction of the tandem duplication, suggesting that the mutant phenotype results from the novel juxtaposition of sequences. We discuss models that relate the tandem duplication of sequences to altered gene expression.


Confocal microscopy for astrocyte in vivo imaging: Recycle and reuse in microscopy.
In vivo imaging is one of the ultimate and fundamental approaches for the study of the brain. Two-photon laser scanning microscopy (2PLSM) constitutes the state-of-the-art technique in current neuroscience to address questions regarding brain cell structure, development and function, blood flow regulation and metabolism. This technique evolved from laser scanning confocal microscopy (LSCM), which impacted the field with a major improvement in image resolution of live tissues in the 1980s compared to widefield microscopy. While nowadays some of the unparalleled features of 2PLSM make it the tool of choice for brain studies in vivo, such as the possibility to image deep within a tissue, LSCM can still be useful in this matter. Here we discuss the validity and limitations of LSCM and provide a guide to perform high-resolution in vivo imaging of the brain of live rodents with minimal mechanical disruption employing LSCM. We describe the surgical procedure and experimental setup that allowed us to record intracellular calcium variations in astrocytes evoked by sensory stimulation, and to monitor intact neuronal dendritic spines and astrocytic processes as well as blood vessel dynamics. Therefore, in spite of certain limitations that need to be carefully considered, LSCM constitutes a useful, convenient, and affordable tool for brain studies in vivo.


The Effect of Nanosizing on the Oxidation of Partially Oxidized Copper Nanoparticles.
Copper nanoparticles are of great interest in various applications, such as catalysis, cooling fluids, conductive inks or for their antibacterial activity. In this paper, the thermal behavior of copper nanoparticles was studied using thermogravimetry, differential thermal analysis and differential scanning calorimetry. Original Cu samples as well as the products of oxidation were analysed by X-ray diffraction, scanning/transmission electron microscopy and energy dispersive spectroscopy. A step-by-step oxidation mechanism during the oxidation of Cu nano-powders was observed. The Cu-nano oxidation starts slightly above 150 °C when bulk copper does not yet react. The dominant oxidation product in the first step is Cu2O while CuO was identified as the final state of oxidation. Our results confirm an easier oxidation process of Cu-nano than Cu-micro particles, which must be attributed to kinetic not thermodynamic aspects of oxidation reactions.


The development of spatial frequency discrimination.
We compared thresholds for discriminating spatial frequency for children aged 5, 7, and 9 years, and adults at two baseline spatial frequencies (1 and 3 cpd). In Experiment 1, the minimum change from baseline necessary to detect a change in spatial frequency from either baseline decreased with age from 34% in 5-year-olds to 11% in 7-year-olds, 8% in 9-year-olds, and 6% in adults. The data were best fit by an exponential function reflecting the rapid improvement in thresholds between 5 and 7 years of age and more gradual improvement thereafter (r(2) = 0.50, p < 0.0001). In Experiment 2, 5-year-olds' thresholds were higher than those of adults, even when memory demands were eliminated by presenting the two spatial frequencies side by side for an unlimited time. The pattern of development for sensitivity to spatial frequency (this study) resembles those for the development of sensitivity to orientation (T. L. Lewis, S. E. Chong, & D. Maurer, 2009) and contrast (D. Ellemberg, T. L. Lewis, C. H. Lui, & D. Maurer, 1999). The similar patterns are consistent with theories of common underlying mechanisms in primary visual cortex (A. Vincent & D. Regan, 1995; W. Zhu, M. Shelley, & R. Shapley, 2008) and suggest that those mechanisms continue to develop throughout childhood.


Adult Neurogenesis Conserves Hippocampal Memory Capacity.
The hippocampus is crucial for declarative memories in humans and encodes episodic and spatial memories in animals. Memory coding strengthens synaptic efficacy via an LTP-like mechanism. Given that animals store memories of everyday experiences, the hippocampal circuit must have a mechanism that prevents saturation of overall synaptic weight for the preservation of learning capacity. LTD works to balance plasticity and prevent saturation. In addition, adult neurogenesis in the hippocampus is proposed to be involved in the down-scaling of synaptic efficacy. Here, we show that adult neurogenesis in male rats plays a crucial role in the maintenance of hippocampal capacity for memory (learning and/or memory formation). Neurogenesis regulated the maintenance of LTP, with decreases and increases in neurogenesis prolonging or shortening LTP persistence, respectively. Artificial saturation of hippocampal LTP impaired memory capacity in contextual fear conditioning, which completely recovered after 14 d, which was the time required for LTP to decay to the basal level. Memory capacity gradually recovered in parallel with neurogenesis-mediated gradual decay of LTP. Ablation of neurogenesis by x-ray irradiation delayed the recovery of memory capacity, whereas enhancement of neurogenesis using a running wheel sped up recovery. Therefore, one benefit of ongoing adult neurogenesis is the maintenance of hippocampal memory capacity through homeostatic renewing of hippocampal memory circuits. Decreased neurogenesis in aged animals may be responsible for the decline in cognitive function with age.SIGNIFICANCE STATEMENT Learning many events each day increases synaptic efficacy via LTP, which can prevent the storage of new memories in the hippocampal circuit. In this study, we demonstrate that hippocampal capacity for the storage of new memories is maintained by ongoing adult neurogenesis through homoeostatic renewing of hippocampal circuits in rats. A decrease or an increase in neurogenesis, respectively, delayed or sped up the recovery of memory capacity, suggesting that hippocampal adult neurogenesis plays a critical role in reducing LTP saturation and keeps the gate open for new memories by clearing out the old memories from the hippocampal memory circuit.


DOES THE KINETICS OF TRYPSIN DIGESTION DEPEND ON THE FORMATION OF A COMPOUND BETWEEN ENZYME AND SUBSTRATE.
1. The velocity of hydrolysis of gelatin by trypsin increases more slowly than the gelatin concentration and finally becomes nearly independent of the gelatin concentration. The relative velocity of hydrolysis of any two substrate concentrations is independent of the quantity of enzyme used to make the comparison. 2. The rate of hydrolysis is independent of the viscosity of the solution. 3. The percentage retardation of the rate of hydrolysis by inhibiting substances, is independent of the substrate concentration. 4. There is experimental evidence that the enzyme and inhibiting substance are combined to form a widely dissociated compound. 5. If the substrate were also combined with the enzyme, an increase in the substrate concentration should affect the equilibrium between the enzyme and the inhibiting substance. This is not the case. 6. The rate of digestion of a mixture of casein and gelatin is equal to the sum of the rates of hydrolysis of the two substances alone, as it should be if the rate is proportional to the concentration of free enzyme. This contradicts the saturation hypothesis. 7. If the reaction is followed by determining directly the change in the substrate concentration, it is found that this change agrees with the law of mass action; i.e., the rate of digestion is proportional to the substrate concentration.


Isolation, structure elucidation, and antitumor activity of spirohexenolides A and B.
In this report, we describe the discovery of a pair of bioactive spirotetronates, spirohexenolides A (1) and B (2), that arose from the application of mutagenesis, clonal selection techniques, and media optimization to strains of Streptomyces platensis. The structures of spirohexenolides A (1) and B (2) were elucidated through X-ray crystallography and confirmed by 1D and 2D NMR studies. Under all examined culture conditions, spirohexenolide A (1) was the major metabolite with traces of spirohexenolide B (2) arising in cultures containing increased loads of adsorbent resins. Spirohexenolide A (1) inhibited tumor cell growth with GI(50) values spanning from 0.1 to 17 microM across the NCI 60 cell line panel. An increased activity was observed in leukemia (GI(50) value of 254 nM in RPMI-8226 cells), lung cancer (GI(50) value of 191 nM in HOP-92 cells), and colon cancer (GI(50) value of 565 nM in SW-620 cells) tumor cells. Metabolite 1 was fluorescent and could be examined on a confocal fluorescent microscope with conventional laser excitation and filter sets. Time lapse imaging studies indicated that spirohexenolide A (1) was readily taken up by tumor cells, appearing through the cell immediately after dosing and subcellularly localizing in the lysosomes. This activity, combined with a unique selectivity in NCI 60 cancer cell line screening, indicates that 1 warrants further chemotherapeutic evaluation.


Multiple in vivo footprints are specific to the active allele of the X-linked human hypoxanthine phosphoribosyltransferase gene 5' region: implications for X chromosome inactivation.
Dosage compensation of X-linked genes in male and female mammals is accomplished by random inactivation of one X chromosome in each female somatic cell. As a result, a transcriptionally active allele and a transcriptionally inactive allele of most X-linked genes reside within each female nucleus. To examine the mechanism responsible for maintaining this unique system of differential gene expression, we have analyzed the differential binding of regulatory proteins to the 5' region of the human hypoxanthine phosphoribosyltransferase (HPRT) gene on the active and inactive X chromosomes. Studies of DNA-protein interactions associated with the transcriptionally active and inactive HPRT alleles were carried out in intact cultured cells by in vivo footprinting by using ligation-mediated polymerase chain reaction and dimethyl sulfate. Analysis of the active allele demonstrates at least six footprinted regions, whereas no footprints were detected on the inactive allele. Of the footprints on the active allele, at least four occur over canonical GC boxes or Sp1 consensus binding sites, one is associated with a potential AP-2 binding site, and another is associated with a DNA sequence not previously reported to interact with a sequence-specific DNA-binding factor. While no footprints were observed for the HPRT gene on the inactive X chromosome, reactivation of the inactive allele with 5-azacytidine treatment restored the in vivo footprint pattern found on the active allele. Results of these experiments, in conjunction with recent studies on the X-linked human PGK-1 gene, bear implications for models of X chromosome inactivation.


The Highland Health Advocates: a preliminary evaluation of a novel programme addressing the social needs of emergency department patients.
Patients commonly come to the emergency department (ED) with social needs. To address this, we created the Highland Health Advocates (HHA), an ED-based help desk and medical-legal partnership using undergraduate volunteers to help patients navigate public resources and provide onsite legal and social work referrals. We were able to provide these services in English and Spanish. We aimed to determine the social needs of the patients who presented to our ED and the potential impact of the programme in resolving those needs and connecting them to a 'medical home' (defined as a consistent, primary source of medical care such as a primary care doctor or clinic). ED patients at a US safety net hospital were enrolled in a 1:2 ratio in a quasi-experiment comparing those who received intervention from the HHA during a limited access rollout with controls who received usual care on days with no help desk. We collected a baseline social needs evaluation, with follow-up assessments at 1 and 6 months. Primary outcomes were linkages for the primary identified need and to a medical home within 1 month. Other outcomes at 6 months included whether a patient (1) felt helped; 2) had a decreased number of ED visits; (3) had the primary identified need met; (4) had a primary doctor; and (5) had a change in self-reported health status. We enrolled 459 subjects (intervention=154, control=305). Housing (41%), employment (23%) and inability to pay bills (22%) were participants' top identified needs. At baseline, 32% reported the ED as their medical home, with the intervention cohort having higher ED utilisation (>1 ED visit in the prior month: 49% vs 24%). At 1 month, 185 (40%) subjects were reached for follow-up, with more HHA subjects linked to a resource (59% vs 37%) and a medical home (92% vs 76%). At 6 months, 75% of subjects felt HHA was helpful and more subjects in the HHA group had a doctor (93% v 69%). No difference was found in ED utilisation, primary need resolution or self-reported health status. Health-related social needs are common in this US safety net ED. Our help desk is one possible model for addressing social needs.


Fatal anaphylactic reaction to intravenous gadobutrol, a gadolinium-based MRI contrast agent.
We present the rare case of a fatal anaphylactic reaction to gadobutrol, a magnetic resonance imaging contrast agent, in a 42-year-old man. The patient underwent elective magnetic resonance imaging for diagnostic clarification of a suspicious finding in the abdomen. The patient had undergone contrast-enhanced computed tomography previously without the occurrence of any adverse effects. Adverse drug reactions in gadobutrol have a very low prevalence of 0.32%-3.5%, with serious adverse drug reactions in <0.1%. There are only a few cases of fatal anaphylactoid reactions to gadolinium-based contrast agents in general. However, if an anaphylactoid reaction occurs, it can present itself with a fulminant course within minutes.