The anatomic evaluation of the internal mammary artery using multidetector CT angiography.
To determine the normal anatomic features and variations of the internal mammary arteries (IMAs) and to describe the relationship between the diameter and distance to the sternal edge of the IMAs, gender, and location (right-left) of the IMAs in patients who underwent multidetector computed tomography (MDCT) angiography of the thorax for various reasons. A total of 164 patients who underwent MDCT angiography of the thoracic vascular structures for various reasons were analyzed retrospectively. The right and left IMAs were analyzed individually, and normal anatomic features and variations were recorded. The relationships between gender, side and diameter of the IMAs, and distance to the sternal edge of the IMAs were evaluated. There were 328 (164 right, 164 left) IMAs in 164 patients (110 males, 54 females; mean age, 43.96 years). A total number of five arteries (1.5%) had anatomic variation. Whereas 325 IMAs had an origin separate from the subclavian artery, three LIMA of the 328 arteries (0.91%) had a common origin with the thyrocervical trunk or costocervical trunk. Two (0.6%) IMAs (one LIMA and one RIMA) in the same patient were duplicated at the level of the first and second costal cartilage. There were no statistically significant correlations between age and diameter or between gender and diameter of the RIMA and LIMA at the origin and level of tracheal bifurcation (P > 0.05). Mean distance between the lateral margin of the sternum and midpoint of LIMA and RIMA were 12.42 mm and 13.00 mm, respectively. The normal anatomic features and variations of the IMAs have an important role in cardiovascular bypass surgery, breast reconstruction, and percutaneous transthoracic procedures. MDCT angiography allows a precise and detailed evaluation of IMAs.


Vaccination with autologous dendritic cells loaded with autologous tumor lysate or homogenate combined with immunomodulating radiotherapy and/or preleukapheresis IFN-α in patients with metastatic melanoma: a randomised "proof-of-principle" phase II study.
Vaccination with dendritic cells (DC) loaded with tumor antigens elicits tumor-specific immune responses capable of killing cancer cells without inducing meaningful side-effects. Patients with advanced melanoma enrolled onto our phase II clinical studies have been treated with autologous DC loaded with autologous tumor lysate/homogenate matured with a cytokine cocktail, showing a clinical benefit (PR + SD) in 55.5% of evaluable cases to date. The beneficial effects of the vaccine were mainly restricted to patients who developed vaccine-specific immune response after treatment. However, immunological responses were only induced in about two-thirds of patients, and treatments aimed at improving immunological responsiveness to the vaccine are needed. This is a phase II, "proof-of-principle", randomized, open-label trial of vaccination with autologous DC loaded with tumor lysate or homogenate in metastatic melanoma patients combined with immunomodulating RT and/or preleukapheresis IFN-α. All patients will receive four bi-weekly doses of the vaccine during the induction phase and monthly doses thereafter for up to a maximum of 14 vaccinations or until confirmed progression. Patients will be randomized to receive:(1.) three daily doses of 8 Gy up to 12 Gy radiotherapy delivered to one non-index metastatic field between vaccine doses 1 and 2 and, optionally, between doses 7 and 8, using IMRT-IMAT techniques;(2.) daily 3 MU subcutaneous IFN-α for 7 days before leukapheresis;(3.) both 1 and 2;(4.) neither 1 nor 2.At least six patients eligible for treatment will be enrolled per arm. Daily 3 MU IL-2 will be administered subcutaneously for 5 days starting from the second day after each vaccine dose. Serial DTH testing and blood sampling to evaluate treatment-induced immune response will be performed. Objective response will be evaluated according to immune-related response criteria (irRC). Based upon the emerging role of radiotherapy as an immunologic modifier, we designed a randomized phase II trial adding radiotherapy and/or preleukapheresis IFN-α to our DC vaccine in metastatic melanoma patients. Our aim was to find the best combination of complementary interventions to enhance anti-tumor response induced by DC vaccination, which could ultimately lead to better survival and milder toxicity.


DNA-dependent protein kinase-independent activation of p53 in response to DNA damage.
Phosphorylation at serine 15 of the human p53 tumor suppressor protein is induced by DNA damage and correlates with accumulation of p53 and its activation as a transcription factor. The DNA-dependent protein kinase (DNA-PK) can phosphorylate serine 15 of human p53 and the homologous serine 18 of murine p53 in vitro. Contradictory reports exist about the requirement for DNA-PK in vivo for p53 activation and cell cycle arrest in response to ionizing radiation. While primary SCID (severe combined immunodeficiency) cells, that have defective DNA-PK, show normal p53 activation and cell cycle arrest, a transcriptionally inert form of p53 is induced in the SCID cell line SCGR11. In order to unambiguously define the role of the DNA-PK catalytic subunit (DNA-PKcs) in p53 activation, we examined p53 phosphorylation in mouse embryonic fibroblasts (MEFs) from DNA-PKcs-null mice. We found a similar pattern of serine 18 phosphorylation and accumulation of p53 in response to irradiation in both control and DNA-PKcs-null MEFs. The induced p53 was capable of sequence-specific DNA binding even in the absence of DNA-PKcs. Transactivation of the cyclin-dependent-kinase inhibitor p21, a downstream target of p53, and the G1 cell cycle checkpoint were also found to be normal in the DNA-PKcs -/- MEFs. Our results demonstrate that DNA-PKcs, unlike the related ATM protein, is not essential for the activation of p53 and G1 cell cycle arrest in response to ionizing radiation.


Membrane interactions of diphtheria toxin analyzed using in vitro synthesized mutants.
We have developed a system to study the interactions of diphtheria toxin with the cell surface using non-toxic mutant proteins synthesized in vitro. Proteins obtained by N-terminal deletions containing the whole B fragment bound strongly to cells. Deletions extending into the B fragment did not yield an autonomous binding domain. Loss of only the N-terminal 3 kd of the B fragment significantly impaired the ability to recognize the receptor. This, together with previous reports that the C-terminal end of the B fragment is required for binding, suggests that both ends of the B fragment are necessary for receptor recognition. Receptor bound diphtheria toxin undergoes a conformational change at pH less than 5.3 that results in translocation of the A fragment to the cytosol and the appearance of a B fragment-derived 25 kd polypeptide (P25) resistant to externally applied protease. Only the B fragment was required for generation of P25. N-terminal deletions of 130 amino acids or more resulted in proteins that gave rise to P25 at higher pH than full length toxin. Furthermore, a second protease-inaccessible polypeptide of 18 kd (P18) was observed.


Altered Global Brain Signal during Physiologic, Pharmacologic, and Pathologic States of Unconsciousness in Humans and Rats.
Consciousness is supported by integrated brain activity across widespread functionally segregated networks. The functional magnetic resonance imaging-derived global brain signal is a candidate marker for a conscious state, and thus the authors hypothesized that unconsciousness would be accompanied by a loss of global temporal coordination, with specific patterns of decoupling between local regions and global activity differentiating among various unconscious states. Functional magnetic resonance imaging global signals were studied in physiologic, pharmacologic, and pathologic states of unconsciousness in human natural sleep (n = 9), propofol anesthesia (humans, n = 14; male rats, n = 12), and neuropathological patients (n = 21). The global signal amplitude as well as the correlation between global signal and signals of local voxels were quantified. The former reflects the net strength of global temporal coordination, and the latter yields global signal topography. A profound reduction of global signal amplitude was seen consistently across the various unconscious states: wakefulness (median [1st, 3rd quartile], 0.46 [0.21, 0.50]) versus non-rapid eye movement stage 3 of sleep (0.30 [0.24, 0.32]; P = 0.035), wakefulness (0.36 [0.31, 0.42]) versus general anesthesia (0.25 [0.21, 0.28]; P = 0.001), healthy controls (0.30 [0.27, 0.37]) versus unresponsive wakefulness syndrome (0.22 [0.15, 0.24]; P < 0.001), and low dose (0.07 [0.06, 0.08]) versus high dose of propofol (0.04 [0.03, 0.05]; P = 0.028) in rats. Furthermore, non-rapid eye movement stage 3 of sleep was characterized by a decoupling of sensory and attention networks from the global network. General anesthesia and unresponsive wakefulness syndrome were characterized by a dissociation of the majority of functional networks from the global network. This decoupling, however, was dominated by distinct neuroanatomic foci (e.g., precuneus and anterior cingulate cortices). The global temporal coordination of various modules across the brain may distinguish the coarse-grained state of consciousness versus unconsciousness, while the relationship between the global and local signals may define the particular qualities of a particular unconscious state.


VCSEL-powered and polarization-maintaining fiber-optic grating vector rotation sensor.
A compact fiber-optic vector rotation sensor in which a short section of polarization-maintaining (PM) fiber stub containing a straight fiber Bragg grating (FBG) is spliced to another single mode fiber without any lateral offset is proposed and experimentally demonstrated. Due to the intrinsic birefringence of the PM fiber, two well-defined resonances (i.e. orthogonally polarized FBG core modes) with wavelength separation of 0.5 nm have been achieved in reflection, and they exhibit a high sensitivity to fiber rotation. Both the orientation and the angle of rotation can be determined unambiguously via simple power detection of the relative amplitudes of the orthogonal core reflections. Meanwhile, instead of using a broadband source (BBS), the sensor is powered by a commercial vertical cavity surface emitting laser (VCSEL) with the laser wavelength matched to the PM-FBG core modes, which enables the sensor to work at much higher power levels (~15 dB better than BBS). This improves the signal-to-noise ratio considerably (~50 dB), and makes a demodulation filter unnecessary. Vector rotation measurement with a sensitivity of 0.09 dB/deg has been achieved via cost-effective single detector real time power measurement, and the unwanted power fluctuations and temperature perturbations can be effectively referenced out.


Nutritional modulation of age-related macular degeneration.
Age-related macular degeneration (AMD) is the leading cause of blindness in the elderly worldwide. It affects 30-50 million individuals and clinical hallmarks of AMD are observed in at least one third of persons over the age of 75 in industrialized countries (Gehrs et al., 2006). Costs associated with AMD are in excess of $340 billion US (American-Health-Assistance-Foundation, 2012). The majority of AMD patients in the United States are not eligible for clinical treatments (Biarnes et al., 2011; Klein et al., 2011). Preventive interventions through dietary modulation are attractive strategies because many studies suggest a benefit of micro- and macronutrients with respect to AMD, as well as other age-related debilities, and with few, if any, adverse effects (Chiu, 2011). Preservation of vision would enhance quality of life for millions of elderly people, and alleviate the personal and public health financial burden of AMD (Frick et al., 2007; Wood et al., 2011). Observational studies indicate that maintaining adequate levels of omega-3 fatty acids (i.e. with 2 servings/week of fish) or a low glycemic index diet may be particularly beneficial for early AMD and that higher levels of carotenoids may be protective, most probably, against neovascular AMD. Intervention trials are needed to better understand the full effect of these nutrients and/or combinations of nutrients on retinal health. Analyses that describe effects of a nutrient on onset and/or progress of AMD are valuable because they indicate the value of a nutrient to arrest AMD at the early stages. This comprehensive summary provides essential information about the value of nutrients with regard to diminishing risk for onset or progress of AMD and can serve as a guide until data from ongoing intervention trials are available.


An experimental and theoretical NMR study of NH-benzimidazoles in solution and in the solid state: proton transfer and tautomerism.
This paper reports the (1)H, (13)C and (15)N NMR experimental study of five benzimidazoles in solution and in the solid state ((13)C and (15)N CPMAS NMR) as well as the theoretically calculated (GIAO/DFT) chemical shifts. We have assigned unambiguously the "tautomeric positions" (C3a/C7a, C4/C7 and C5/C6) of NH-benzimidazoles that, in some solvents and in the solid state, appear different (blocked tautomerism). In the case of 1H-benzimidazole itself we have measured the prototropic rate in HMPA-d 18.


Age-dependent expression of DNMT1 and DNMT3B in PBMCs from a large European population enrolled in the MARK-AGE study.
Aging is associated with alterations in the content and patterns of DNA methylation virtually throughout the entire human lifespan. Reasons for these variations are not well understood. However, several lines of evidence suggest that the epigenetic instability in aging may be traced back to the alteration of the expression of DNA methyltransferases. Here, the association of the expression of DNA methyltransferases DNMT1 and DNMT3B with age has been analysed in the context of the MARK-AGE study, a large-scale cross-sectional study of the European general population. Using peripheral blood mononuclear cells, we assessed the variation of DNMT1 and DNMT3B gene expression in more than two thousand age-stratified women and men (35-75 years) recruited across eight European countries. Significant age-related changes were detected for both transcripts. The level of DNMT1 gradually dropped with aging but this was only observed up to the age of 64 years. By contrast, the expression of DNMT3B decreased linearly with increasing age and this association was particularly evident in females. We next attempted to trace the age-related changes of both transcripts to the influence of different variables that have an impact on changes of their expression in the population, including demographics, dietary and health habits, and clinical parameters. Our results indicate that age affects the expression of DNMT1 and DNMT3B as an almost independent variable in respect of all other variables evaluated.


Cardiovascular disease competes with breast cancer as the leading cause of death for older females diagnosed with breast cancer: a retrospective cohort study.
Many women who survive breast cancer die of causes unrelated to their cancer diagnosis. This study was undertaken to assess factors that are related to breast cancer mortality versus mortality from other causes and to describe the leading causes of death among older women diagnosed with breast cancer. Women diagnosed with breast cancer at age 66 or older between 1992 and 2000 were identified in the Surveillance, Epidemiology and End Results-Medicare linked database and followed through the end of 2005. A total of 63,566 women diagnosed with breast cancer met the inclusion criteria and were followed for a median of approximately nine years. Almost one-half (48.7%) were alive at the end of follow-up. Ages and comorbidities at the time of diagnosis had the largest effects on mortality from other causes, while tumor stage, tumor grade, estrogen receptor status, age and comorbidities at the time of diagnosis all had effects on breast cancer-specific mortality. Fully adjusted relative hazards of the effects of comorbidities on breast cancer-specific mortality were 1.24 (95% confidence interval (95% CI) 1.13 to 1.26) for cardiovascular disease, 1.13 (95% CI 1.13 to 1.26) for previous cancer, 1.13 (95% CI 1.05 to 1.22) for chronic obstructive pulmonary disease and 1.10 (95% CI 1.03 to 1.16) for diabetes. Among the total study population, cardiovascular disease was the primary cause of death in the study population (15.9% (95% CI 15.6 to 16.2)), followed closely by breast cancer (15.1% (95% CI 14.8 to 15.4)). Comorbid conditions contribute importantly to both total mortality and breast cancer-specific mortality among breast cancer survivors. Attention to reducing the risk of cardiovascular disease should be a priority for the long-term care of women following the diagnosis and treatment of breast cancer.


Evidence-based tailoring of behavior-change campaigns: increasing fluoride-free water consumption in rural Ethiopia with persuasion.
Two hundred million people worldwide are at risk of developing dental and skeletal fluorosis due to excessive fluoride uptake from their water. Since medical treatment of the disease is difficult and mostly ineffective, preventing fluoride uptake is crucial. In the Ethiopian Rift Valley, a fluoride-removal community filter was installed. Despite having access to a fluoride filter, the community used the filter sparingly. During a baseline assessment, 173 face-to-face interviews were conducted to identify psychological factors that influence fluoride-free water consumption. Based on the results, two behavior-change campaigns were implemented: a traditional information intervention targeting perceived vulnerability, and an evidence-based persuasion intervention regarding perceived costs. The interventions were tailored to household characteristics. The campaigns were evaluated with a survey and analyzed in terms of their effectiveness in changing behavior and targeted psychological factors. While the intervention targeting perceived vulnerability showed no desirable effects, cost persuasion decreased the perceived costs and increased the consumption of fluoride-free water. This showed that altering subjective perceptions can change behavior even without changing objective circumstances. Moreover, interventions are more effective if they are based on evidence and tailored to specific households.


Environmental Interactions and Epistasis Are Revealed in the Proteomic Responses to Complex Stimuli.
Ultimately, the genotype of a cell and its interaction with the environment determine the cell's biochemical state. While the cell's response to a single stimulus has been studied extensively, a conceptual framework to model the effect of multiple environmental stimuli applied concurrently is not as well developed. In this study, we developed the concepts of environmental interactions and epistasis to explain the responses of the S. cerevisiae proteome to simultaneous environmental stimuli. We hypothesize that, as an abstraction, environmental stimuli can be treated as analogous to genetic elements. This would allow modeling of the effects of multiple stimuli using the concepts and tools developed for studying gene interactions. Mirroring gene interactions, our results show that environmental interactions play a critical role in determining the state of the proteome. We show that individual and complex environmental stimuli behave similarly to genetic elements in regulating the cellular responses to stimuli, including the phenomena of dominance and suppression. Interestingly, we observed that the effect of a stimulus on a protein is dominant over other stimuli if the response to the stimulus involves the protein. Using publicly available transcriptomic data, we find that environmental interactions and epistasis regulate transcriptomic responses as well.


Sex and race or ethnicity disparities in opioid prescriptions for dental diagnoses among patients receiving Medicaid.
The objective of this study was to identify specific factors (sex, race or ethnicity, and health care provider type) associated with patient receipt of an opioid prescription after a dental diagnosis. The authors used Medicaid claims dated from January 1, 2013, through September 30, 2015, for 13 U.S. states in this study. The authors identified oral health-related conditions by using International Classification of Diseases, Ninth Revision, Clinical Modification diagnosis codes 520.0 through 529.9. During the 2013 through 2015 study period, among the more than 1,008,400 Medicaid patients with a dental diagnosis, 19.8% filled an opioid prescription within 14 days of diagnosis. Female patients were 50% more likely to receive an opioid prescription for pain management of a dental condition than were men (odds ratio [OR], 1.50; 95% confidence interval [CI], 1.49 to 1.52). Non-Hispanic whites and African Americans were approximately twice as likely to receive opioids than were Hispanics (OR, 2.12; 95% CI, 2.05 to 2.19; OR, 1.90; 95% CI, 1.84 to 1.96, respectively). Patients receiving oral health care in an emergency department were more than 7 times more likely to receive an opioid prescription than were patients treated in a dental office (OR, 7.28; 95% CI, 7.13 to 7.43). Patients with a dental condition diagnosed were more than 4 times as likely to receive an opioid from a nurse practitioner as from a dentist (OR, 4.31; 95% CI, 4.19 to 4.44). Opioid use was substantially higher among African American female patients (OR, 2.02; 95% CI, 1.93 to 2.10) and non-Hispanic white female patients (OR, 2.16; 95% CI, 2.07 to 2.24) than among Hispanic female patients. Opioid prescribing patterns differ depending on patient race or ethnicity, sex, and health care provider source in patients with a dental diagnosis in the United States. Dentists are providing substantially fewer opioid prescriptions compared with their medical colleagues for pain treatment after a dental diagnosis in the Medicaid population. When considering pain management for dental and related conditions, dentists should continue with conservative prescribing practices as recommended.


The role of intravascular ultrasound in the management of spontaneous coronary artery dissection.
Primary or spontaneous coronary artery dissection (SCAD) is an unusual but increasingly recognized cause of acute myocardial ischemia and sudden cardiac death. Typically, SCAD presents in younger patients without conventional risk factors for coronary artery disease. It occurs more commonly in women than in men, and frequently during pregnancy or the postpartum period. Its pathophysiology is poorly understood, and there is considerable controversy regarding the optimal management of patients with SCAD-related myocardial ischemia. Therapeutic approaches include conservative medical therapy, coronary artery bypass graft surgery and percutaneous coronary intervention (PCI). We present four cases of SCAD to illustrate specific aspects of the presentation and management of this condition, with particular reference to the importance of intravascular ultrasound (IVUS) to aid diagnosis and guide subsequent PCI.


High power fiber delivery for laser ignition applications.
The present contribution provides a concise review of high power fiber delivery research for laser ignition applications. The fiber delivery requirements are discussed in terms of exit energy, intensity, and beam quality. Past research using hollow core fibers, solid step-index fibers, and photonic crystal and bandgap fibers is summarized. Recent demonstrations of spark delivery using large clad step-index fibers and Kagome photonic bandgap fibers are highlighted.


Persistent low levels of human chorionic gonadotropin: A premalignant gestational trophoblastic disease.
This study was undertaken to evaluate the significance of persistent low-level human chorionic gonadotropin (hCG) titers (usually <50 IU/L) in the absence of clinical evidence of pregnancy or gestational trophoblastic disease. The USA hCG Reference Service consulted on 114 cases with persistent low levels of hCG; 51 had false-positive hCG results. The remaining 63 cases had real hCG results and are presented here. Antecedent gestational events included hydatidiform mole (27), pregnancy (35), and gestational trophoblastic neoplasm (1). Forty of the 63 (64%) cases received therapy, including chemotherapy (38), hysterectomy (2), or both (10). Despite treatment, in all cases, low hCG titers persisted. After 1 to 4.5 years of low titers, four women had a sudden rapid increase in hCG levels, and malignant disease was confirmed or clearly suggested (gestational trophoblastic neoplasm [3] and placental site trophoblastic tumor [1]). Invasive trophoblast antigen (ITA) is a marker of invasive cytotrophoblast cells. ITA was measured in 38 of the cases with persistent low hCG, in all cases ITA accounted for less than 25% of the hCG concentration. It was also determined in the 4 cases indicated with malignant disease, accounting for more than 80% of the hCG. The presence of persistent low-level hCG titers defines a subset of women with preinvasive or quiescent gestational trophoblastic disease. ITA effectively detected the presence or absence of invasive cells in these cases. The recommended management of the quiescent disease is close surveillance without therapy until malignant disease detected.


Association of microsatellite markers near the fibrillin 1 gene on human chromosome 15q with scleroderma in a Native American population.
To localize disease genes for scleroderma, or systemic sclerosis (SSc), in a population of Choctaw Native Americans with a high prevalence of SSc, in which there is evidence of a possible founder effect. A candidate gene approach was used in which microsatellite alleles on human chromosomes 15q and 2q, homologous to the murine tight skin 1 (tsk1) and tsk2 loci, respectively, were analyzed in Choctaw SSc cases and race-matched normal controls for possible disease association. Genotyping first-degree relatives of the cases identified potential disease haplotypes, and haplotype frequencies were obtained by expectation-maximization and maximum-likelihood estimation methods. Simultaneously, the ancestral origins of contemporary Choctaw SSc cases were ascertained using census and historical records. A multilocus 2-cM haplotype was identified on human chromosome 15q homologous to the murine tsk1 region, which showed a significantly increased frequency in SSc cases compared with controls. This haplotype contains 2 intragenic markers for the fibrillin 1 (FBN1) gene. Genealogical studies demonstrated that the SSc cases were distantly related, and their ancestry could be traced back to 5 founding families in the mid-eighteenth century. The probability that the SSc cases share this haplotype due to familial aggregation effects alone was calculated and found to be very low. There was no evidence of any microsatellite allele disturbances on chromosome 2q in the region homologous to the tsk2 locus or the region containing the interleukin-1 family. A 2-cM haplotype on chromosome 15q that contains FBN1 is associated with scleroderma in Choctaw Native Americans from Oklahoma. This haplotype may have been inherited from common founders about 10 generations ago and may contribute to the high prevalence of SSc that is now seen.


Carbon Nanohybrids as Electro-Responsive Drug Delivery Systems.
Electro-responsive nanomaterials are usually made with polyelectrolytes able to undergo shrinkage or swelling by tuning on electrical fields. Nevertheless, the electrical conductivity of many polymeric materials used for the fabrication of release devices is not high enough to achieve an effective modulation of the drug release. The incorporation of conducting materials (e.g. carbon nanostructures) in polymeric networks has been proposed as a valuable strategy to overcome this limitation. In this regard, carbon nanotubes and graphene, by virtue of their unique chemical structures and attractive physiochemical properties, have been receiving exciting attention primarily in biology and medicine. By their incorporation into composite hydrogels, the biocompatibility and biodegradability of polymers can be merged with the favorable properties of carbon nanostructures, such as enhanced cellular uptake, electromagnetic, and magnetic behavior. The applicability of carbon hybrid materials to modulate release of therapeutics in response to an external current voltage, is being extensively investigated in the present review.


The molecular basis for modulation of human Vγ9Vδ2 T cell responses by CD277/butyrophilin-3 (BTN3A)-specific antibodies.
Human Vγ9Vδ2 T cells are well known for their rapid and potent response to infection and tumorigenesis when in the presence of endogenous or exogenous phosphoisoprenoids. However, the molecular mechanisms behind the activation of this γδ T cell population remains unclear. Evidence pointing to a role for the CD277/butyrophilin-3 (BTN3A) molecules in this response led us to investigate the structures of these molecules and their modifications upon binding to an agonist antibody (20.1) that mimics phosphoisoprenoid-mediated Vγ9Vδ2 activation and an antagonist antibody (103.2) that inhibits this reactivity. We find that the three BTN3A isoforms: BTN3A1, BTN3A2, and BTN3A3, have high structural homology to the B7 superfamily of proteins and exist as V-shaped homodimers in solution, associating through the membrane proximal C-type Ig domain. The 20.1 and 103.2 antibodies bind to separate epitopes on the BTN3A Ig-V domain with high affinity but likely with different valencies based on their binding orientation. These structures directly complement functional studies of this system that demonstrate that BTN3A1 is necessary for Vγ9Vδ2 activation and begin to unravel the extracellular events that occur during stimulation through the Vγ9Vδ2 T cell receptor.


The Danish oral health care service for children: its achievements.
Child dental services in Denmark were legally defined in 1971 and have functioned under this law for about a decade. Although no operational goals were defined, it is fair to say that the activities and results of the service have lived up to the expectations held at the inception of the law. The evaluation and planning system has made it possible to monitor developments in health as well as the use of resources. There have been consistent trends towards improved dental health in children and at the same time no increases in the use of personnel. There is general agreement in society that child services should remain a public activity. Pressures upon the services in the years to come must be expected to mount due to increased competition for scarce resources, simultaneous improvement in dental health and a reduction in the number of children. These pressures will stimulate a re-examination of goals within the service so that appropriate short and long term local action can be taken before outside influences force changes with deleterious effects on the structure and function of the service. The success of the service and its ultimate achievements, therefore, will have to be measured against its abilities to adapt to changes in health as well as to an economic situation which will be more demanding than hitherto.


How interdisciplinary is nanotechnology?
Facilitating cross-disciplinary research has attracted much attention in recent years, with special concerns in nanoscience and nanotechnology. Although policy discourse has emphasized that nanotechnology is substantively integrative, some analysts have countered that it is really a loose amalgam of relatively traditional pockets of physics, chemistry, and other disciplines that interrelate only weakly. We are developing empirical measures to gauge and visualize the extent and nature of interdisciplinary interchange. Such results speak to research organization, funding, and mechanisms to bolster knowledge transfer. In this study, we address the nature of cross-disciplinary linkages using "science overlay maps" of articles, and their references, that have been categorized into subject categories. We find signs that the rate of increase in nano research is slowing, and that its composition is changing (for one, increasing chemistry-related activity). Our results suggest that nanotechnology research encompasses multiple disciplines that draw knowledge from disciplinarily diverse knowledge sources. Nano research is highly, and increasingly, integrative-but so is much of science these days. Tabulating and mapping nano research activity show a dominant core in materials sciences, broadly defined. Additional analyses and maps show that nano research draws extensively upon knowledge presented in other areas; it is not constricted within narrow silos.


Ancient aqueous environments at Endeavour crater, Mars.
Opportunity has investigated in detail rocks on the rim of the Noachian age Endeavour crater, where orbital spectral reflectance signatures indicate the presence of Fe(+3)-rich smectites. The signatures are associated with fine-grained, layered rocks containing spherules of diagenetic or impact origin. The layered rocks are overlain by breccias, and both units are cut by calcium sulfate veins precipitated from fluids that circulated after the Endeavour impact. Compositional data for fractures in the layered rocks suggest formation of Al-rich smectites by aqueous leaching. Evidence is thus preserved for water-rock interactions before and after the impact, with aqueous environments of slightly acidic to circum-neutral pH that would have been more favorable for prebiotic chemistry and microorganisms than those recorded by younger sulfate-rich rocks at Meridiani Planum.


Prevalence of hepatitis B and C seromarkers and abnormal liver function tests among hemophiliacs in Guilan (northern province of Iran).
The aim of this study was to determine the seroprevalence of hepatitis B and C viruses and abnormal liver function tests among hemophiliacs in Guilan province, Iran. Patients registered with Guilan Hemophilia Society (GHS) were enrolled in this study, and evaluated for the presence of hepatitis B surface antigen (HBsAg), hepatitis C virus antibody (HCV-Ab) and liver function tests. One hundred and one patients (100 males, 1 female, mean age 19.7 years, range 3-71 years) from 110 registered hemophiliacs were enrolled in this study. 29 patients (28.7%) had elevated Alanine aminotransferase (ALT), 27 (26.7%) and 72 (71.3%) were positive for HBsAg and HCV-Ab, respectively. Seropositivity for HCV-Ab correlated with the duration of treatment with clotting factor until 1997 (P=0.01). There was also an inverse correlation between factor VIII & IX activity and seropositivity for HCV-Ab (P<0.001) and HBsAg (P<0.04). HCV-Ab was more likely to be positive among those received lyophilized factor VIII and cryoprecipitate than lyophilized factor VIII alone (P<0.01). In addition HCV-Ab seropositivity was more common among those received factor IX concentrate and fresh frozen plasma (FFP) than those received only FFP (P<0.01). Based upon our finding, prevalence of HBsAg positive cases in Guilan province was higher than other studies. Although frequency of HCV-Ab was similar to other studies, frequency of increased ALT was less. Upon the results emerged from this study, we recommend that all hemophiliacs should be vaccinated against HBV and should have regular program for checking HCV.


Activation of human muscle pyruvate dehydrogenase with activity and immobilization.
The activity of pyruvate dehydrogenase (PDH) was measured in samples of triceps muscle obtained in 15 males, before and after immobilization for 5 weeks and 5 months of strength training carried out in random order with cross-over of treatment. Although the total PDH activity was unchanged at 300 nmol min-1 g-1, the proportion in the active form increased after strength training (62 +/- 10.1%) and decreased after immobilization (12 +/- 2.3%) compared with control (36 +/- 3.4%). In six subjects measurements were repeated after 10 min maximal exercise (arm ergometer). The proportion of PDH in the active form increased least after immobilization (to 52 +/- 13.6%), compared with control (95 +/- 11.7%) and post-training (98 +/- 6.6%). After exercise muscle glycogen fell to the greatest extent and lactate rose the least in the post-training state, with opposite findings post-immobilization, suggesting that PDH activation contributes to the control of lactate formation in muscle during heavy exercise, and that the effects of training and immobilization are mediated at least in part through changes in the activation of this regulatory enzyme.


Simulating hypoxia and modelling the airway.
Apnoea due to airway obstruction is an ever present concern in anaesthesia and critical care practice and results in rapid development of hypoxaemia that is not always remediable by manual bag-mask ventilation. As it is often difficult or impossible to study experimentally (although some historical animal data exist), it is useful to model the kinetics of hypoxaemia following airway obstruction. Despite being a complex event, the consequences of airway obstruction can be predicted with reasonable fidelity using mathematical and computer modelling. Over the last 15 years, a number of high fidelity mathematical and computer models have been developed, that have thrown light on this important event.


Clinical potential of sodium-glucose cotransporter 2 inhibitors in the management of type 2 diabetes.
The kidney plays an important role in glucose metabolism, and has been considered a target for therapeutic intervention. The sodium-glucose cotransporter type 2 (SGLT2) mediates most of the glucose reabsorption from the proximal renal tubule. Inhibition of SGLT2 leads to glucosuria and provides a unique mechanism to lower elevated blood glucose levels in diabetes. The purpose of this review is to explore the physiology of SGLT2 and discuss several SGLT2 inhibitors which have clinical data in patients with type 2 diabetes. We performed a PubMed search using the terms "SGLT2" and "SGLT2 inhibitor" through April 10, 2012. Published articles, press releases, and abstracts presented at national and international meetings were considered. SGLT2 inhibitors correct a novel pathophysiological defect, have an insulin-independent action, are efficacious with glycosylated hemoglobin reduction ranging from 0.5% to 1.5%, promote weight loss, have a low incidence of hypoglycemia, complement the action of other antidiabetic agents, and can be used at any stage of diabetes. They are generally well tolerated. However, due to side effects, such as repeated urinary tract and genital infections, increased hematocrit, and decreased blood pressure, appropriate patient selection for drug initiation and close monitoring after initiation will be important. Results of ongoing clinical studies of the effect of SGLT2 inhibitors on diabetic complications and cardiovascular safety are crucial to determine the risk-benefit ratio. A recent decision by the Committee for Medicinal Products for Human Use of the European Medicines Agency has recommended approval of dapagliflozin for the treatment of type 2 diabetes as an adjunct to diet and exercise, in combination with other glucose-lowering medicinal products, including insulin, and as a monotherapy for metformin-intolerant patients. Clinical research also remains to be carried out on the long-term effects of glucosuria and other potential effects of SGLT2 inhibitors, especially in view of the observed increase in the incidence of bladder and breast cancer. SGLT2 inhibitors represent a promising approach for the treatment of diabetes, and could potentially be an addition to existing therapies.


NTP Toxicology and Carcinogenesis Studies of Tetrakis(hydroxymethyl)phosphonium sulfate (THPS) (CAS No. 55566-30-8) and Tetrakis(hydroxymethyl)phosphonium chloride (THPC) (CAS No. 124-64-1) in F344/N Rats and B6C3F1 Mice (Gavage Studies).
Toxicology and carcinogenesis studies of tetrakis(hydroxymethyl)phosphonium sulfate (THPS) and tetrakis(hydroxymethyl)phosphonium chloride (THPC) were conducted because of the widespread use of these chemicals as flame retardants in cotton fabrics. THPS was available as a 72% aqueous solution and THPC as a 75% aqueous solution. Short-term gavage studies with a range of doses were conducted first to identify toxic effects and affected sites and to determine doses for the 2-year studies. The doses selected for the 14-day studies ranged from 12.5 to 200 mg/kg THPS for rats and mice, 9.4 to 150 mg/kg THPC for rats, and 18.8 to 300 mg/kg THPC for mice. Mortality and reduction in body weight gain occurred at the two highest doses in the 14-day studies. There was hind limb paralysis in some rats and mice dosed at the highest concentrations of THPS and THPC. In the 13-week studies, doses of THPS ranged from 5 to 60 mg/kg in rats and from 2 to 180 mg/kg in mice; doses of THPC ranged from 3.75 to 60 mg/kg in rats and from 1.5 to 135 mg/kg in mice. Mortality and reduction in body weight gain occurred at the two higher doses for both sexes and species. Vascular degeneration of hepatocytes or hepatocellular necrosis was a common histopathologic finding. Hind limb paralysis was noted in rats and mice receiving the highest dose of THPC, and axonal degeneration, characterized by swollen axon sheaths, missing or fragmented axons, and some proliferation of neurolemma cells, was observed in rats. These lesions were found in the sciatic nerve, dorsal roots of the caudal spinal nerves, and the tracts of the spinal cord, particularly in the dorsal column of the lumbar cord. Two-year studies were conducted in F344/N rats by administering 0, 5, or 10 mg/kg THPS or 0, 3.75, or 7.5 mg/kg THPC in deionized water by gavage to groups of 49 or 50 animals of each sex, 5 days per week for 103 or 104 weeks. Groups of 49 or 50 B6C3F1 mice were administered 0, 5, or 10 mg/kg THPS (each sex), 0, 7.5, or 15 mg/kg THPC (males), or 0, 15, or 30 mg/kg THPC (females). Survival of male rats was reduced for the low dose (after week 102) and the high dose (after week 67) groups given THPS compared with that of the vehicle controls; survival at terminal kill was as follows: vehicle control, 28/50; low dose, 13/50; high dose, 16/50. Survival of the high dose group of female rats given THPC was lower after week 70 than that of the vehicle controls (survival at terminal kill: 37/50; 34/50; 21/50). Mean body weights of rats dosed with THPS or THPC were comparable to those of the vehicle controls. There was no difference in survival or mean body weights between the vehicle controls and mice dosed with either THPS or THPC. No neurotoxicity or any other signs of clinical toxicity were observed. A nonneoplastic effect common to 13-week and 2-year exposure to THPS or THPC was an increase in the incidence of hepatocellular lesions, primarily cytoplasmic vacuolization. The incidences of this lesion in the two-year studies were dose related for all studies except for the mice receiving THPS. Other lesions observed included focal hyperplasia of the adrenal medulla in high dose male mice given THPS and follicular cell hyperplasia of the thyroid gland in high dose female mice given THPC. The increased incidences of hematopoietic system lesions observed in these studies were not considered biologically related to chemical exposure because the increases were marginal, no dose-response relationship was observed, and the incidences of these lesions are highly variable in untreated rats and mice. The incidences of mononuclear cell leukemia in low dose male rats administered THPS or THPC were somewhat greater than those in the vehicle controls (THPS: 30/50; 36/50; 20/50; THPC: 19/50; 25/50; 16/50). Low dose male mice administered THPS had an increased incidence of malignant lymphomas when compared with vehicle controls (2/50; 9/50; 0/50). These marginal increases in the incidences of hematopoietic system tumors were not considered related to chemical exposure, since they were signific were significant only by the life table tests and were not dose related. THPC demonstrated no mutagenic activity in Salmonella typhimurium strains TA98, TA100, TA1535, or TA1537 with or without metabolic activation. Both THPS and THPC induced forward mutations in mouse lymphoma L5178Y cells without metabolic activation; neither was tested in the presence of S9. THPC increased the frequency of sister-chromatid exchanges and chromosomal aberrations in Chinese hamster ovary cells in the presence and absence of exogenous metabolic activation. An audit of the experimental data was conducted for the 2-year studies of THPS and THPC. No discrepancies were found that influenced the final interpretations. Under the conditions of these 2-year gavage studies, there was no evidence of carcinogenicity of THPS in either sex of F344/N rats or B6C3F1 mice given 5 or 10 mg/kg. There was no evidence of carcinogenicity of THPC in either sex of F344/N rats given 3.75 or 7.5 mg/kg, in male B6C3F1 mice given 7.5 or 15 mg/kg, or in female B6C3F1 mice given 15 or 30 mg/kg.


Mediterranean diet and mortality in Switzerland: an alpine paradox?
Reports on the protective effect of a Mediterranean diet on mortality usually refer to populations from Mediterranean countries, leaving uncertain whether really diet is the fundamental cause. Our aim was to examine the effect of a Mediterranean diet on mortality in Switzerland, a country combining cultural influences from Mediterranean and Central European countries within a common national health and statistical registry. In this prospective investigation, we included 17,861 men and women aged ≥16 years who participated 1977-1993 in health studies and were followed up for survival until 2008 by anonymous record linkage with the Swiss National Cohort. A 9-point score Mediterranean Diet Score (MDS) was used to assess adherence to a Mediterranean diet. Mortality hazard ratios (HR) and 95% confidence intervals (CIs) were calculated by using Cox regression models adjusted for age, sex, survey wave, marital status, smoking, body mass index, language region and nationality. In all language regions, MDS was inversely associated with mortality. Consumption of dairy products was also consistently associated with lower mortality. When categorizing dairy food consumption as beneficial instead of harmful, this association between MDS and mortality increased in strength and was partly statistically significant. For all causes of death combined (HR for a one-point increase in MDS 0.96, 95% CI 0.94-0.98), in men (0.94, 0.92-0.97), in women (0.98, 0.95-1.02) for cardiovascular diseases (CVD, 0.96, 0.92-0.99; 0.95, 0.90-1.00; 0.98, 0.92-1.04) and for cancer (0.95, 0.92-0.99; 0.92, 0.88-0.97; 0.98, 0.93-1.04). Stronger adherence to a Mediterranean diet was associated with lower all-cause, CVD and cancer mortality, largely independently of cultural background. These associations were primary due to the effect in men. Our finding of a beneficial rather than a deleterious impact of dairy products consumption prompts at considering culturally adapted Mediterranean diet recommendations. However, results should be interpreted with caution since only a crude 1-day dietary estimate was available to assess individuals' habitual dietary intake.


Identification of the epigenetic reader CBX2 as a potential drug target in advanced prostate cancer.
While localized prostate cancer (PCa) can be effectively cured, metastatic disease inevitably progresses to a lethal state called castration-resistant prostate cancer (CRPC). Emerging evidence suggests that aberrant epigenetic repression by the polycomb group (PcG) complexes fuels PCa progression, providing novel therapeutic opportunities. In the search for potential epigenetic drivers of CRPC, we analyzed the molecular profile of PcG members in patient-derived xenografts and clinical samples. Overall, our results identify the PcG protein and methyl-lysine reader CBX2 as a potential therapeutic target in advanced PCa. We report that CBX2 was recurrently up-regulated in metastatic CRPC and that elevated CBX2 expression was correlated with poor clinical outcome in PCa cohorts. Furthermore, CBX2 depletion abrogated cell viability and induced caspase 3-mediated apoptosis in metastatic PCa cell lines. Mechanistically explaining this phenotype, microarray analysis in CBX2-depleted cells revealed that CBX2 controls the expression of many key regulators of cell proliferation and metastasis. Taken together, this study provides the first evidence that CBX2 inhibition induces cancer cell death, positioning CBX2 as an attractive drug target in lethal CRPC.


Improving the prognosis of health care in the USA.
Although health care expenditure per capita is higher in the USA than in any other country, more than 37 million Americans do not have health insurance, and 41 million more have inadequate access to care. Efforts are ongoing to repeal the Affordable Care Act which would exacerbate health-care inequities. By contrast, a universal system, such as that proposed in the Medicare for All Act, has the potential to transform the availability and efficiency of American health-care services. Taking into account both the costs of coverage expansion and the savings that would be achieved through the Medicare for All Act, we calculate that a single-payer, universal health-care system is likely to lead to a 13% savings in national health-care expenditure, equivalent to more than US$450 billion annually (based on the value of the US$ in 2017). The entire system could be funded with less financial outlay than is incurred by employers and households paying for health-care premiums combined with existing government allocations. This shift to single-payer health care would provide the greatest relief to lower-income households. Furthermore, we estimate that ensuring health-care access for all Americans would save more than 68 000 lives and 1·73 million life-years every year compared with the status quo.


Tocotrienol (unsaturated vitamin E) suppresses degranulation of mast cells and reduces allergic dermatitis in mice.
In this study, we examined whether tocotrienol (T3) reduces allergic dermatitis in mice and suppresses degranulation of mast cells. First, allergic dermatitis was examined in the atopic dermatitis model NC/Nga mouse. Allergic dermatitis was induced using picryl chloride in mice with and without administration of T3 (1 mg/day/mouse). Increases in scratching behavior, dermal thickening, and the serum histamine level were greatly reduced in mice treated with T3, indicating that T3 reduces allergic dermatitis in vivo. Next, the effect of T3 on degranulation of mast cells was examined, since these cells release bioactive substances such as histamine. T3 significantly suppressed degranulation of mast cells and significantly reduced histamine release. The effect of T3 on protein kinase C (PKC) activity was also measured, since suppression of this activity may be associated with the mechanism underlying the antidegranulation effect of T3. T3 significantly suppressed PKC activity. Therefore, we conclude that T3 suppresses degranulation of mast cells and reduces allergic dermatitis in mice through reduction of PKC activity.


Vitamin D and coronavirus disease 2019 (COVID-19): rapid evidence review.
The rapid global spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus that causes coronavirus disease 2019 (COVID-19), has re-ignited interest in the possible role of vitamin D in modulation of host responses to respiratory pathogens. Indeed, vitamin D supplementation has been proposed as a potential preventative or therapeutic strategy. Recommendations for any intervention, particularly in the context of a potentially fatal pandemic infection, should be strictly based on clinically informed appraisal of the evidence base. In this narrative review, we examine current evidence relating to vitamin D and COVID-19 and consider the most appropriate practical recommendations. Although there are a growing number of studies investigating the links between vitamin D and COVID-19, they are mostly small and observational with high risk of bias, residual confounding, and reverse causality. Extrapolation of molecular actions of 1,25(OH)2-vitamin D to an effect of increased 25(OH)-vitamin D as a result of vitamin D supplementation is generally unfounded, as is the automatic conclusion of causal mechanisms from observational studies linking low 25(OH)-vitamin D to incident disease. Efficacy is ideally demonstrated in the context of adequately powered randomised intervention studies, although such approaches may not always be feasible. At present, evidence to support vitamin D supplementation for the prevention or treatment of COVID-19 is inconclusive. In the absence of any further compelling data, adherence to existing national guidance on vitamin D supplementation to prevent vitamin D deficiency, predicated principally on maintaining musculoskeletal health, appears appropriate.


Usefulness of a program of hospital-supervised physical training in patients with cystic fibrosis.
Exercise is an important part of normal childhood, but the ability to exercise may be impaired in chronic lung diseases such as cystic fibrosis (CF). Improving exercise performance by training is very attractive. The aim of the present study was the evaluation of the effects of a physical aerobic training program, performed in the Children's Hospital and Research Institute "Bambino Gesù" (Rome, Italy) in outpatient CF children, supervised by a physician. Twelve patients (mean forced expiratory flow in 1 sec (FEV1), 71%), age range 12-24 years (16.7 +/- 4.4 years), were enrolled. They performed a maximal exercise stress test on the treadmill (modified Bruce protocol) with breath-by-breath determination of oxygen consumption (VO2) to maximum at end-exercise; we measured time of exercise (TE), maximal heart rate (Hrmax) in beats per minute (bpm), and maximal systolic blood pressure (SBPm) in mmHg. The program consisted of 12 weeks of training twice a week. Each training session consisted of walking or running on the treadmill for 30 min at the speed that allowed the child to attain 60% of the maximal heart rate obtained during a baseline stress test for 4 weeks, 70% in the following 4 weeks, and 80% in the last 4 weeks, under strict medical supervision. HR was continously monitored. There was no change in FEV1 and forced vital capacity after the treatment period. Hrmax and SBPm also remained the same (P = 0.37 and P = 0.25, respectively). There was a significant increase in TE (P < 0.002), VO2, VO2/kg, and pulmonary ventilation (VE) (P < 0.0001, P < 0.001, and P < 0.001, respectively). This pilot study showed that a simple training program improves short-term cardiopulmonary fitness in children with CF. Further studies with a larger sample and for a more prolonged time are necessary to assess if sport can have a long-term effect on lung function or survival in CF patients.


ATPase inhibitors of heat-shock protein 90, second season.
In the past four years, the ATP-dependent heat-shock protein 90 has remained the focus of much interest. Phase I and phase II anticancer clinical trials with first-generation inhibitors, although sometimes disappointing, have yet to report a forbidding side-effect inherent to the inhibition of this chaperone, which has a very complex and widespread role in cell biochemistry. Research in the field has started to unravel an elaborate regulation picture leading to the proper folding of many proteins. On the medicinal chemistry side, a second wave of inhibitors has been reported. This review attempts to describe all the ATPase inhibitors of HSP90 reported since our last survey.


Square-wave retarder for polarization computer-generated holography.
An etched calcite square-wave retarder is designed, fabricated, and demonstrated as an illuminator for an interlaced polarization computer-generated hologram (PCGH). The calcite square-wave retarder enables alternating columns of orthogonal linear polarizations to illuminate the interlaced PCGH. Together, these components produce a speckled, tangentially polarized PCGH diffraction pattern with a measured ratio of polarization of 84% and a degree of linear polarization of 0.81. An experimental alignment tolerance analysis is also reported.


Involvement of Type IV Pili in Pathogenicity of Plant Pathogenic Bacteria.
Type IV pili (T4P) are hair-like appendages found on the surface of a wide range of bacteria belonging to the β-, γ-, and δ-Proteobacteria, Cyanobacteria and Firmicutes. They constitute an efficient device for a particular type of bacterial surface motility, named twitching, and are involved in several other bacterial activities and functions, including surface adherence, colonization, biofilm formation, genetic material uptake and virulence. Tens of genes are involved in T4P synthesis and regulation, with the majority of them being generally named pil/fim genes. Despite the multiple functionality of T4P and their well-established role in pathogenicity of animal pathogenic bacteria, relatively little attention has been given to the role of T4P in plant pathogenic bacteria. Only in recent years studies have begun to examine with more attention the relevance of these surface appendages for virulence of plant bacterial pathogens. The aim of this review is to summarize the current knowledge about T4P genetic machinery and its role in the interactions between phytopathogenic bacteria and their plant hosts.


The prevalence of intra-uterine growth retardation in patients with positive contraction stress tests.
The gestational ages of 66 neonates delivered after positive contraction stress tests (CSTs) were estimated by means of the Dubowitz score. Forty-three (65%) of these infants had birth weights below the 10th percentile according to their gestational age. At the beginning of the study 40% of the infants were found to be growth-retarded, but as experience with antenatal fetal heart rate monitoring increased the CSTs were interpreted more accurately. This led to an increase in the incidence of IUGR in infants born after a positive CST to 82%, a predictive value which means that the test compares favourably with other methods of diagnosing the small-for-dates fetus.


Controlling urban air pollution caused by households: uncertainty, prices, and income.
We examine the control of air pollution caused by households burning wood for heating and cooking in the developing world. Since the problem is one of controlling emissions from nonpoint sources, regulations are likely to be directed at household choices of wood consumption and combustion technologies. Moreover, these choices are subtractions from, or contributions to, the pure public good of air quality. Consequently, the efficient policy design is not independent of the distribution of household income. Since it is unrealistic to assume that environmental authorities can make lump sum income transfers part of control policies, efficient control of air pollution caused by wood consumption entails a higher tax on wood consumption and a higher subsidy for more efficient combustion technologies for higher income households. Among other difficulties, implementing a policy to promote the adoption of cleaner combustion technologies must overcome the seemingly paradoxical result that efficient control calls for higher technology subsidies for higher income households.


Tobacco smoking and gingival health in a Saudi Arabian population.
While cigarette smoking is recognized as being detrimental to periodontal health, the effect of water pipe smoking on gingival health is not known. The present study was conducted to determine whether water pipe smoking has an influence on gingival health. A cross-sectional study was carried out in 244 individuals aged 25-70 years. The levels of plaque and gingivitis were recorded on four sites of all present teeth, using the plaque index (PI) and gingival index (GI). Information about oral hygiene practices, dental care and smoking habits was obtained at the time of the clinical examination in accordance with a predetermined questionnaire. The means of plaque index and gingival index values were 1.2 and 0.9, respectively. Similarly, the mean percentages of surfaces with plaque and gingival bleeding sites were 66.7% and 30.4%, respectively. There was an overall significant association between smoking and plaque index and gingival index (F = 22.9 and F = 10.8, respectively, p < 0.001). Oral hygiene was inferior in water pipe smokers, cigarette smokers, and mixed smokers when compared to non-smokers. The correlation between plaque % and gingival bleeding % in cigarette smokers was significantly weaker than in non-smokers. It was also weaker in water pipe smokers, but the difference was not statistically significant. The gingival bleeding response to plaque was significantly suppressed in cigarette smokers. There was a tendency towards suppression also in water pipe smokers.


Estimation of displacement derivatives in digital holographic interferometry using a two-dimensional space-frequency distribution.
The paper introduces a two-dimensional space-frequency distribution based method to directly obtain the unwrapped estimate of the phase derivative which corresponds to strain in digital holographic interferometry. In the proposed method, a two-dimensional pseudo Wigner-Ville distribution of the reconstructed interference field is evaluated and the peak of the distribution provides information about the phase derivative. The presence of a two-dimensional window provides high robustness against noise and enables simultaneous measurement of phase derivatives along both spatial directions. Simulation and experimental results are presented to demonstrate the method's applicability for phase derivative estimation.


Effects of combined flocculant - Lanthanum modified bentonite treatment on aquatic macroinvertebrate fauna.
A low dose flocculant (FeCl3), combined with lanthanum modified bentonite (LMB) as phosphate-binding agent, has been applied for eutrophication management in Lake De Kuil (The Netherlands). After the treatment, the state of the lake shifted from hypertrophic to mesotrophic. Although macroinvertebrate fauna is important for lake ecosystems, the knowledge of its response to this lake restoration method is fragmented and scarce. Because insight in the macroinvertebrate fauna response is important to assess future applications, pre and post application macroinvertebrate assemblages were identified in Lake De Kuil. The research was accompanied by a microcosm experiment in which the effects of LMB, FeCl3 and LMB + FeCl3 were studied on macroinvertebrate communities. Results show the reduction of macroinvertebrate numbers and taxa during the first month following the application. The number of Gastropoda was strikingly reduced one month after the application. One year after the application, the macroinvertebrate numbers and taxa exceeded the pre-application situation and Gastropoda and Oligochaeta prospered. The effects one month after the treatment are most likely due to the combination of physical impacts of the use of bentonite and chemical impacts of the use of FeCl3, while effects after one year are likely attributed to the shift in trophic state of the lake.


Recruitment of N-CoR/SMRT-TBLR1 corepressor complex by unliganded thyroid hormone receptor for gene repression during frog development.
The corepressors N-CoR (nuclear receptor corepressor) and SMRT (silencing mediator for retinoid and thyroid hormone receptors) interact with unliganded nuclear hormone receptors, including thyroid hormone (T(3)) receptor (TR). Several N-CoR/SMRT complexes containing histone deacetylases have been purified. The best studied among them are N-CoR/SMRT complexes containing TBL1 (transducin beta-like protein 1) or TBLR1 (TBL1-related protein). Despite extensive studies of these complexes, there has been no direct in vivo evidence for the interaction of TBL1 or TBLR1 with TR or the possible involvement of such complexes in gene repression by any nuclear receptors in any animals. Here, we used the frog oocyte system to demonstrate that unliganded TR interacts with TBLR1 and recruits TBLR1 to its chromatinized target promoter in vivo, accompanied by histone deacetylation and gene repression. We further provide evidence to show that the recruitment of TBLR1 or related proteins is important for repression by unliganded TR. To investigate the potential role for TBLR1 complexes during vertebrate development, we made use of T(3)-dependent amphibian metamorphosis as a model. We found that TBLR1, SMRT, and N-CoR are recruited to T(3)-inducible promoters in premetamorphic tadpoles and are released upon T(3) treatment, which induces metamorphosis. More importantly, we demonstrate that the dissociation of N-CoR/SMRT-TBLR1 complexes from endogenous TR target promoters is correlated with the activation of these genes during spontaneous metamorphosis. Taken together, our studies provide in vivo evidence for targeted recruitment of N-CoR/SMRT-TBLR1 complexes by unliganded TR in transcriptional repression during vertebrate development.


Abnormalities of the inactive X chromosome are a common feature of BRCA1 mutant and sporadic basal-like breast cancer.
As a clinical entity, breast cancer appears to be a series of subforms, each with a relatively specific molecular phenotype. Among the characteristics that differentiate these subforms are sex hormone receptor expression, HER2 expression, p53 mutation, high-grade histopathology, and particular gene expression array patterns. Sporadic basal-like breast cancer is one such form. It is a relatively common, high-grade, hormone receptor and HER2-expression-negative, p53 mutation-bearing tumor and is particularly lethal. Although wild type for BRCA1, it is a sporadic phenocopy of most cases of BRCA1(/) breast cancer. Not only do the cells of the two tumors resemble one another with respect to the above-noted characteristics, they also share a defect in the maintenance of an intact, inactive X chromosome (Xi). Other high-grade and most low-grade tumors are rarely defective at Xi. This evidence suggests that an Xi defect contributes to the evolution of both sporadic and BRCA1(/) basal-like breast tumors.


Can we assess the model complexity for a bioprocess: theory and example of the anaerobic digestion process.
In this paper we propose a methodology to determine the structure of the pseudo-stoichiometric coefficient matrix K in a mass balance based model, i.e. the maximal number of biomasses that must be taken into account to reproduce an available data set. It consists in estimating the number of reactions that must be taken into account to represent the main mass transfer within the bioreactor. This provides the dimension of K. The method is applied to data from an anaerobic digestion process and shows that even a model including a single biomass is sufficient. Then we apply the same method to the "synthetic data" issued from the complex ADM1 model, showing that the main model features can be obtained with two biomasses.


A Cycloaddition Approach toward the Synthesis of Substituted Indolines and Tetrahydroquinolines.
The intramolecular Diels-Alder reaction of 2-substituted aminofurans (IMDAF) results in the formation of various indolines and tetrahydroquinolines. The isolation of these ring systems from the IMDAF reaction can be rationalized in terms of an initial [4 + 2]-cycloaddition that first produces an oxa-bridged cycloadduct, which was not detected since it readily underwent nitrogen-assisted ring opening. Proton exchange followed by an eventual dehydration provides the aromatic product. In certain cases, the intermediate cyclohexadienol can be isolated and independently converted to the final product in high yield. The starting 2-aminofurans were readily prepared from furanyl acyl azide by a Curtius rearrangement in the presence of an alcohol. Alkylation of the resulting N-alkyl carbamate with an alkenyl bromide allows for the synthesis of a wide variety of cycloaddition precursors. The scope of the IMDAF reaction was evaluated by using mono- and disubstituted alkenes, electron rich and electron deficient olefins, and acetylenic tethers. Cyclic 2-amidofurans were also synthesized using a related intramolecular Diels-Alder reaction of 2-amido-substituted oxazoles which contain a tethered alkyne. This transformation represents a new route to this rare heterocyclic ring system. The sequential cycloaddition method was used for a formal synthesis of the pyrrolophenanthridone alkaloid hippadine.


Concurrent thrombotic thrombocytopenic purpura and immune thrombocytopenic purpura in a patient with metastatic neuroendocrine tumour successfully treated with rituximab-CVP.
We report a case of concurrent thrombotic thrombocytopenic purpura (TTP) and immune thrombocytopenic purpura (ITP) in a 63-year-old woman who had been receiving treatment with bevacizumab for metastatic neuroendocrine tumour (NET). At diagnosis, she had severe anaemia and thrombocytopenia with elevated lactate dehydrogenase and many schistocytes on the peripheral blood smear. She was treated with frequent fresh frozen plasma infusions and plasmapheresis with poor response. Later, she was found to have platelet surface glycoprotein antibodies in the serum and received four cycles of rituximab, vincristine, cyclophosphamide (rituximab-CVP) and steroids in addition to plasma therapy. The haemoglobin and platelet counts improved. To our knowledge, this is the first reported case of concurrent TTP and ITP in a patient with metastatic NET diagnosed while receiving bevacizumab therapy, who was successfully treated with the combination of rituximab, vincristine, cyclophosphamide and steroids.


SU-E-I-62: Investigation of Dominant Factors Affecting Fatigue in Image Reading of Radiologists.
To investigate dominant factors affecting fatigue in image reading of radiologists. Two kinds of fatigue were assessed in this study. One was fatigue in the central nervous system evaluated by the critical fusion frequency (CFF). The other was eye fatigue evaluated by a score determined from a questionnaire based on the oculomotor strain subscale from the Simulator Sickness Questionnaire (SSQ). When fatigue increases, the CFF and the SSQ score indicate low and high values, respectively. The fatigue of seventeen radiologists was assessed before and after their daily image reading. The reading times and the numbers of images were different among the assessments, and ranged about 1.5 - 5.0 hours and 1,000 - 12,000 images, respectively. The assessments of fatigue were repeated four times for each radiologist on different days. Finally, the measurements of the two kinds of fatigue were analyzed in terms of years of experience, age, sleeping time the previous night, ambient light conditions, reading time, and the numbers of interpreted images, series, and cases. The CFF and SSQ score after image reading were significantly lower and higher than those measured before image reading, respectively. Younger and less experienced radiologists indicated a higher level of fatigue than older and more experienced radiologists in both the CFF and the SSQ score. When radiologists interpreted clinical images for longer hours, the SSQ score tended to be higher. On the other hand, there was little incremental difference in the CFF among different lengths of reading time. No obvious differences were observed in the other items. Less experience with reading images, a younger age, and a longer reading time could be dominant factors affecting fatigue in image reading.


A novel Rad24 checkpoint protein complex closely related to replication factor C.
Rad24 functions in the DNA damage checkpoint pathway of Saccharomyces cerevisiae. Here, analysis of Rad24 in whole cell extracts demonstrated that its mass was considerably greater than its predicted molecular weight, suggesting that Rad24 is a component of a protein complex. The Rad24 complex was purified to homogeneity. In addition to Rad24, the complex included polypeptides of 40 kDa and 35 kDa. The 40 kDa species was found by mass spectrometry to contain Rfc2 and Rfc3, subunits of replication factor C (RFC), a five subunit protein that is required for the loading of polymerases onto DNA during replication and repair [3]. We hypothesised that other RFC subunits, all of which share sequence homologles with Rad24, might also be components of the Rad24 complex. Reciprocal co-immunoprecipitation studies were performed using extracts prepared from strains containing epitope-tagged RFC proteins. These experiments showed that the small RFC proteins, Rfc2, Rfc3, Rfc4 and Rfc5, interacted with Rad24, whereas the Rfc1 subunit did not. We suggest that this RFC-like Rad24 complex may function as a structure-specific sensor in the DNA damage checkpoint pathway.


Bone metabolic responses to low energy availability achieved by diet or exercise in active eumenorrheic women.
We aimed to explore the effects of low energy availability (EA)[15 kcal·kg lean body mass (LBM)-1·d-1] achieved by diet or exercise on bone turnover markers in active, eumenorrheic women. By using a crossover design, ten eumenorrheic women (VO2 peak: 48.1 ± 3.3 ml·kg-1·min-1) completed all three, 3-day conditions in a randomised order: controlled EA (CON; 45 kcal·kgLBM-1·d-1), low EA through dietary energy restriction (D-RES; 15 kcal·kgLBM-1·d-1) and low EA through increasing exercise energy expenditure (E-RES; 15 kcal·kgLBM-1·d-1), during the follicular phase of three menstrual cycles. In CON, D-RES and E-RES, participants consumed diets providing 45, 15 and 45 kcal·kgLBM-1·d-1. In E-RES only, participants completed supervised running sessions (129 ± 10 min·d-1) at 70% of their VO2 peak that resulted in an exercise energy expenditure of 30 kcal·kg LBM-1·d-1. Blood samples were collected at baseline (BASE) and at the end of the 3-day period (D6) and analysed for bone turnover markers (β-CTX and P1NP), markers of calcium metabolism (PTH, albumin-adjusted Ca, Mg and PO4) and hormones (IGF-1, T3, insulin, leptin and 17β-oestradiol). In D-RES, P1NP concentrations at D6 decreased by 17% (BASE: 54.8 ± 12.7 μg·L-1, D6: 45.2 ± 9.3 μg·L-1, P < 0.001, d = 0.91) and were lower than D6 concentrations in CON (D6: 52.5 ± 11.9 μg·L-1, P = 0.001). P1NP did not change significantly in E-RES (BASE: 55.3 ± 14.4 μg·L-1, D6: 50.9 ± 15.8 μg·L-1, P = 0.14). β-CTX concentrations did not change following D-RES (BASE: 0.48 ± 0.18 μg·L-1, D6: 0.55 ± 0.17 μg·L-1) or E-RES (BASE: 0.47 ± 0.24 μg·L-1, D6: 0.49 ± 0.18 μg·L-1) (condition × time interaction effect, P = 0.17). There were no significant differences in P1NP (P = 0.25) or β-CTX (P = 0.13) responses between D-RES and E-RES. Both conditions resulted in reductions in IGF-1 (-13% and - 23% from BASE in D-RES and E-RES, both P < 0.01) and leptin (-59% and - 61% from BASE in D-RES and E-RES, both P < 0.001); T3 decreased in D-RES only (-15% from BASE, P = 0.002) and PO4 concentrations decreased in E-RES only (-9%, P = 0.03). Low EA achieved through dietary energy restriction resulted in a significant decrease in bone formation but no change in bone resorption, whereas low EA achieved through exercise energy expenditure did not significantly influence bone metabolism. Both low EA conditions elicited significant and similar changes in hormone concentrations.


Edema is not a reliable diagnostic sign to exclude small brain metastases.
No prior systematic study on the extent of vasogenic edema (VE) in patients with brain metastases (BM) exists. Here, we aim to determine 1) the general volumetric relationship between BM and VE, 2) a threshold diameter above which a BM shows VE, and 3) the influence of the primary tumor and location of the BM in order to improve diagnostic processes and understanding of edema formation. This single center, retrospective study includes 173 untreated patients with histologically proven BM. Semi-manual segmentation of 1416 BM on contrast-enhanced T1-weighted images and of 865 VE on fluid-attenuated inversion recovery/T2-weighted images was conducted. Statistical analyses were performed using a paired-samples t-test, linear regression/generalized mixed-effects model, and receiver-operating characteristic (ROC) curve controlling for the possible effect of non-uniformly distributed metastases among patients. For BM with non-confluent edema (n = 545), there was a statistically significant positive correlation between the volumes of the BM and the VE (P < 0.001). The optimal threshold for edema formation was a diameter of 9.4 mm for all BM. The primary tumors as interaction term in multivariate analysis had a significant influence on VE formation whereas location had not. Hence VE development is dependent on the volume of the underlying BM and the site of the primary neoplasm, but not from the location of the BM.


Differential expression of 2',3'-cyclic-nucleotide 3'-phosphodiesterase and neural lineage markers correlate with glioblastoma xenograft infiltration and patient survival.
Glioblastoma multiforme (GBM) is a poorly treated human brain cancer with few established clinically useful molecular prognostic markers. We characterized glioblastoma stem-like cells (GSC) according to developmental neural lineage markers and correlated their expression with patient survival. Immunoblot array of neural lineage markers classified five independently isolated human GSC lines into three classes exhibiting differential expression of oligodendrocyte progenitor cells (OPC), astrocyte progenitor cells (APC), and neural progenitor cells (NPC) markers. Immunodeficient mice were orthotopically implanted with each cell line to evaluate tumor infiltration and recipient survival. 2',3'-Cyclic-nucleotide 3'-phosphodiesterase (CNP) antigenic expression was used to evaluate a clinically annotated GBM tissue microarray with 115 specimens. We report that molecular classification of patient-derived GSCs using neural lineage markers show association with differential xenograft invasiveness, and also show significant correlation to survival in both the mouse model and human patients. Orthotopic implantation into immunodeficient mice showed Ki-67 proliferative index independent xenograft infiltration: class I GSCs (OPC and NPC positive) established focal lesions, class II GSCs (NPC positive) formed minimally invasive lesions, and class III GSCs (APC positive) established highly infiltrative lesions. The OPC marker, CNP also exhibited high expression in focal xenografts versus low expression in invasive xenografts. Differential CNP expression correlated with mouse model survival, and CNP immunoassay of a large GBM tissue microarray also showed significant differential patient survival. GSC classification with developmental neural lineage markers revealed CNP as a novel and potentially useful clinical prognosis marker, and suggests clinical importance for patient-specific GSC analysis.


Physical activity and postmenopausal breast cancer risk in the NIH-AARP diet and health study.
Although physical activity has been associated with reduced breast cancer risk, whether this association varies across breast cancer subtypes or is modified by reproductive and lifestyle factors is unclear. We examined physical activity in relation to postmenopausal breast cancer risk in 182,862 U.S. women in the NIH-AARP Diet and Health Study. Physical activity was assessed by self-report at baseline (1995-1996), and 6,609 incident breast cancers were identified through December 31, 2003. Cox regression was used to estimate the relative risk (RR) and 95% confidence interval (95% CI) of postmenopausal breast cancer overall and by tumor characteristics. Effect modification by select reproductive and lifestyle factors was also explored. In multivariate models, the most active women experienced a 13% lower breast cancer risk versus inactive women (RR, 0.87; 95% CI, 0.81-0.95). This inverse relation was not modified by tumor stage or histology but was suggestively stronger for estrogen receptor (ER)-negative (RR, 0.75; 95% CI, 0.54-1.04) than ER-positive (RR, 0.97; 95% CI, 0.84-1.12) breast tumors and was suggestively stronger for overweight/obese (RR, 0.86; 95% CI, 0.77-0.96) than lean (RR, 0.95; 95% CI, 0.87-1.05) women. The inverse relation with physical activity was also more pronounced among women who had never used menopausal hormone therapy and those with a positive family history of breast cancer than their respective counterparts. Physical activity was associated with reduced postmenopausal breast cancer risk, particular to ER-negative tumors. These results, along with heterogeneity in the physical activity-breast cancer relation for subgroups of menopausal hormone therapy use and adiposity, indicate that physical activity likely influences breast cancer risk via both estrogenic and estrogen-independent mechanisms.


Lessons learned from recruiting socioeconomically disadvantaged smokers into a pilot randomized controlled trial to explore the role of Exercise Assisted Reduction then Stop (EARS) smoking.
Research is needed on what influences recruitment to smoking reduction trials, and how to increase their reach. The present study aimed to i) assess the feasibility of recruiting a disadvantaged population, ii) examine the effects of recruitment methods on participant characteristics, iii) identify resource requirements for different recruitment methods, and iv) to qualitatively assess the acceptability of recruitment. This was done as part of a pilot two-arm trial of the effectiveness of a novel behavioral support intervention focused on increasing physical activity and reducing smoking, among disadvantaged smokers not wishing to quit. Smokers were recruited through mailed invitations from three primary care practices (62 participants) and one National Health Stop Smoking Service (SSS) database (31 participants). Six other participants were recruited via a variety of other community-based approaches. Data were collected through questionnaires, field notes, work sampling, and databases. Chi-squared and t-tests were used to compare baseline characteristics of participants. We randomized between 5.1 and 11.1% of those invited through primary care and SSS, with associated researcher time to recruit one participant varying from 18 to 157 minutes depending on time and intensity invested.Only six participants were recruited through a wide variety of other community-based approaches, with an associated researcher time of 469 minutes to recruit one participant. Targets for recruiting a disadvantaged population were met, with 91% of the sample in social classes C2 to E (NRS social grades, UK), and 41% indicating mental health problems. Those recruited from SSS were more likely to respond to an initial letter, had used cessation aids before, and had attempted to quit in the past year. Overall, initial responders were more likely to be physically active than those who were recruited via follow-up telephone calls. No other demographics or behaviour characteristics were associated with recruitment approach or intensity of effort. Qualitative feedback indicated that participants had been attracted by the prospect of support that focused on smoking reduction rather than abrupt quitting. Mailed invitations, and follow-up, from health professionals was an effective method of recruiting disadvantaged smokers into a trial of an exercise intervention to aid smoking reduction. Recruitment via community outreach approaches was largely ineffective. ISRCTN identifier: 13837944 , registered on 6 July 2010.


Endodontic Treatment in Switzerland. A National Survey
The purpose of this study was to collect information about current treatment protocols of endodontic procedures among general practitioners (GP), endodontically interested practitioners (EI) and endodontic specialists (ES) in Switzerland and to compare them with available endodontic quality guidelines and former surveys. A questionnaire was distributed to 1,522 attendees of two national conferences (Swiss Dental Association, 2015; Swiss Society for Endodontology, 2016) addressing socio-demographic characteristics and specific questions about root canal treatments (RCT) including techniques and materials used. Five hundred and six surveys were collected comprising data of 81.8% GP, 14.4% EI and 3.8% ES (response rate: 33.2%). The majority of respondents was male (59.9%), 40–59 years old (55.9%) with >20 years of clinical experience (54.3%). 498 participants (98.4%) stated to perform RCT. Rubber dam was always used in 52.2% of GP, 69.9% of EI and 89.5% of ES, while never-user accounted for 14.1%. Overall, 63.9% used loupes and 13.7% microscopes (mostly by ES). Rotating instruments were selected in 43.2%, followed by conventional hand-instruments (37.8%) and reciprocating instruments (19%). GP rarely activated irrigants and applied NaOCl in lower concentrations (>0.5–1%) compared to ES. GP preferred lateral compaction (57%), ES performed vertical compaction in 89.5%. 92% completed RCT after two or more visits. The majority of dentists in Switzerland follow the available quality guidelines and the present findings are coherent with internationally published surveys. Nonetheless, discrepancies are still present between daily practice and endodontic quality guidelines, especially with the routine use of rubber dam and working length determination, predominantly within the endodontic non-specialists.


Combining thermal and visible imagery for estimating canopy temperature and identifying plant stress.
Thermal imaging is a potential tool for estimating plant temperature, which can be used as an indicator of stomatal closure and water deficit stress. In this study, a new method for processing and analysing thermal images was developed. By using remote sensing software, the information from thermal and visible images was combined, the images were classified to identify leaf area and sunlit and shaded parts of the canopy, and the temperature statistics for specific canopy components were calculated. The method was applied to data from a greenhouse water-stress experiment of Vicia faba L. and to field data for Vitis vinifera L. Vaseline-covered and water-sprayed plants were used as dry and wet references, respectively, and two thermal indices, based on temperature differences between the canopy and reference surfaces, were calculated for single Vicia faba plants. The thermal indices were compared with measured stomatal conductance. The temperature distributions of sunlit and shaded leaf area of Vitis vinifera canopies from natural rainfall and irrigation treatments were compared. The present method provides two major improvements compared with earlier methods for calculating thermal indices. First, it allows more accurate estimation of the indices, which are consequently more closely related to stomatal conductance. Second, it gives more accurate estimates of the temperature distribution of the shaded and sunlit parts of canopy, and, unlike the earlier methods, makes it possible to quantify the relationship between temperature variation and stomatal conductance.


N-Atom transfer via thermal or photolytic activation of a Co-azido complex with a PNP pincer ligand.
Thermal or photolytic activation of well-defined mononuclear [Co(N3)(PNP)] (PNP = 2,2'-bis(diisopropylphosphino)-4,4'-ditolylamido) results in the structurally characterized dinuclear species [Co(μ-N;κ3-P,N,N-PNPN)]2 (3), with two N-bridging phosphiniminato bridgeheads. Density Functional Theory (DFT) calculations indicate the intermediacy of a mononuclear cobalt-nitrido complex, followed by N-migratory insertion into a Co-P bond. Reaction of 3 with two equiv. HCl leads to rupture of the dimer with formation of mononuclear [CoCl(PNPNH)] (4) by protonation of the N-bridges.


Identification of novel gene expression targets for the Ras association domain family 1 (RASSF1A) tumor suppressor gene in non-small cell lung cancer and neuroblastoma.
RASSF1A is a recently identified 3p21.3 tumor suppressor gene. The high frequency of epigenetic inactivation of this gene in a wide range of human sporadic cancers including non-small cell lung cancer (NSCLC) and neuroblastoma suggests that RASSF1A inactivation is important for tumor development. Although little is known about the function of RASSF1A, preliminary data suggests that it may have multiple functions. To gain insight into RASSF1A functions in an unbiased manner, we have characterized the expression profile of a lung cancer cell line (A549) transfected with RASSF1A. Initially we demonstrated that transient expression of RASSF1A into the NSCLC cell line A549 induced G(1) cell cycle arrest, as measured by propidium iodide staining. Furthermore, annexin-V staining showed that RASSF1A-expressing cells had an increased sensitivity to staurosporine-induced apoptosis. We then screened a cDNA microarray containing more than 6000 probes to identify genes differentially regulated by RASSF1A. Sixty-six genes showed at least a 2-fold change in expression. Among these were many genes with relevance to tumorigenesis involved in transcription, cytoskeleton, signaling, cell cycle, cell adhesion, and apoptosis. For 22 genes we confirmed the microarray results by real-time RT-PCR and/or Northern blotting. In silico, we were able to confirm the majority of these genes in other NSCLC cell lines using published data on gene expression profiles. Furthermore, we confirmed 10 genes at the RNA level in two neuroblastoma cell lines, indicating that these RASSF1A target genes have relevance in non-lung cell backgrounds. Protein analysis of six genes (ETS2, Cyclin D3, CDH2, DAPK1, TXN, and CTSL) showed that the changes induced by RASSF1A at the RNA level correlated with changes in protein expression in both non-small cell lung cancer and neuroblastoma cell lines. Finally, we have used a transient assay to demonstrate the induction of CDH2 and TGM2 by RASSF1A in NSCLC cell lines. We have identified several novel targets for RASSF1A tumor suppressor gene both at the RNA and the protein levels in two different cellular backgrounds. The identified targets are involved in diverse cellular processes; this should help toward understanding mechanisms that contribute to RASSF1A biological activity.


Soluble Thy-1 reverses lung fibrosis via its integrin-binding motif.
Loss of Thy-1 expression in fibroblasts correlates with lung fibrogenesis; however, the clinical relevance of therapeutic targeting of myofibroblasts via Thy-1-associated pathways remains to be explored. Using single (self-resolving) or repetitive (nonresolving) intratracheal administration of bleomycin in type 1 collagen-GFP reporter mice, we report that Thy-1 surface expression, but not mRNA, is reversibly diminished in activated fibroblasts and myofibroblasts in self-resolving fibrosis. However, Thy-1 mRNA expression is silenced in lung with nonresolving fibrosis following repetitive bleomycin administration, associated with persistent activation of αv integrin. Thy1-null mice showed progressive αv integrin activation and myofibroblast accumulation after a single dose of bleomycin. In vitro, targeting of αv integrin by soluble Thy-1-Fc (sThy-1), but not RLE-mutated Thy-1 or IgG, reversed TGF-β1-induced myofibroblast differentiation in a dose-dependent manner, suggesting that Thy-1's integrin-binding RGD motif is required for the reversibility of myofibroblast differentiation. In vivo, treatment of established fibrosis induced either by single-dose bleomycin in WT mice or by induction of active TGF-β1 by doxycycline in Cc10-rtTA-tTS-Tgfb1 mice with sThy-1 (1000 ng/kg, i.v.) promoted resolution of fibrosis. Collectively, these findings demonstrate that sThy-1 therapeutically inhibits the αv integrin-driven feedback loop that amplifies and sustains fibrosis.


Heat stress and sudden infant death syndrome--stress gene expression after exposure to moderate heat stress.
The aim of the present study was to investigate stress gene expression in cultured primary fibroblasts established from Achilles tendons collected during autopsies from sudden infant death syndrome (SIDS) cases, and age-matched controls (infants dying in a traumatic event). Expression of 4 stress responsive genes, HSPA1B, HSPD1, HMOX1, and SOD2, was studied by quantitative reverse transcriptase PCR analysis of RNA purified from cells cultured under standard or various thermal stress conditions. The expression of all 4 genes was highly influenced by thermal stress in both SIDS and control cells. High interpersonal variance found in the SIDS group indicated that they represented a more heterogeneous group than controls. The SIDS group responded to thermal stress with a higher expression of the HSPA1B and HSPD1 genes compared to the control group, whereas no significant difference was observed in the expression of SOD2 and HMOX1 between the two groups. The differences were related to the heat shock treatment as none of the genes were expressed significantly different in SIDS at base levels at 37 °C. SOD2 and HMOX1 were up regulated in both groups, for SOD2 though the expression was lower in SIDS at all time points measured, and may be less related to heat stress. Being found dead in the prone position (a known risk factor for SIDS) was related to a lower HSPA1B up-regulation in SIDS compared to SIDS found on their side or back. The study demonstrates the potential usefulness of gene expression studies using cultured fibroblasts established from deceased individuals as a tool for molecular and pathological investigations in forensic and biomedical sciences.


Effects of cyclophotocoagulation with a neodymium:yttrium-aluminum-garnet laser on corneal sensitivity, intraocular pressure, aqueous tear production, and corneal nerve morphology in eyes of dogs.
To determine effects of cyclophotocoagulation via administration of 100 J with a neodymium:yttrium aluminum garnet (Nd:YAG) laser on corneal touch threshold (CTT), intraocular pressure (IOP), aqueous tear production, and corneal nerve morphology in eyes of dogs. 15 dogs. Noncontact Nd:YAG laser was transsclerally applied (10 applications; 25 W for 0.1 seconds for each application to each of 4 quadrants) to the ciliary body of the left eye of 15 dogs; the right eye was the control eye. Corneal integrity, CTT, tear production as measured by the Schirmer tear test (STT), and IOP were evaluated for 14 days following laser treatment. On day 14, dogs were euthanatized, eyes harvested, and corneas stained with gold chloride. Major nerve bundles were analyzed by use of a drawing tube attached to a light microscope, and maximum diameters were measured by use of image analysis software. All laser-treated eyes had significantly higher CTT values, compared with control eyes. Six of 15 laser-treated eyes developed ulcerative keratitis. On most days, IOP was significantly lower in laser-treated eyes in both morning and evening. Laser-treated eyes had a significant decrease of approximately 1 nerve bundle/corneal quadrant. Values for STT or nerve bundle diameters did not differ significantly. Administration of 100 J with a Nd:YAG laser effectively reduced IOP while increasing CTT and caused a significant decrease in number, but not diameter, of major corneal nerve bundles. Nerve damage and corneal hypoesthesia are etiologic factors in ulcerative keratitis following Nd:YAG cyclophotocoagulation.


TALEs from a spring--superelasticity of Tal effector protein structures.
Transcription activator-like effectors (TALEs) are DNA-related proteins that recognise and bind specific target sequences to manipulate gene expression. Recently determined crystal structures show that their common architecture reveals a superhelical overall structure that may undergo drastic conformational changes. To establish a link between structure and dynamics in TALE proteins we have employed coarse-grained elastic-network modelling of currently available structural data and implemented a force-probe setup that allowed us to investigate their mechanical behaviour in computer experiments. Based on the measured force-extension curves we conclude that TALEs exhibit superelastic dynamical properties allowing for large-scale global conformational changes along their helical axis, which represents the soft direction in such proteins. For moderate external forcing the TALE models behave like linear springs, obeying Hooke's law, and the investigated structures can be characterised and compared by a corresponding spring constant. We show that conformational flexibility underlying the large-scale motions is not homogeneously distributed over the TALE structure, but instead soft spot residues around which strain is accumulated and which turn out to represent key agents in the transmission of conformational motions are identified. They correspond to the RVD loop residues that have been experimentally determined to play an eminent role in the binding process of target DNA.


Depression and marital status determine the 10-year (2004-2014) prognosis in patients with acute coronary syndrome: the GREECS study.
The aim of the present work was to examine the association of depression and marital status, with the long-term prognosis of acute coronary syndrome (ACS), among a Greek sample of cardiac patients. From October 2003 to September 2004, a sample of 2172 consecutive ACS patients from 6 hospitals was enrolled. In 2013-2014, the 10-year follow-up was performed in 1918 participants. Depressive symptoms were evaluated using the validated CES-D score (range 0-60), while marital status was classified as: single, married/cohabitants, divorced and widowed at the time of entry to the study. Patients in the upper tertile of CES-D (>20 score) had 41% (95%CI 14%, 75%) higher risk of ACS incidence as compared with those in the lowest (<7 score). In contrary, married patients had 29% lower risk (95%CI 6%, 46%) of ACS mortality compared with single, widowed or divorced. Multi-adjusted analysis revealed that among the 'not married' patients, 1-point increase in the CES-D score was associated with 2% (p = .02) and 4% (p = .001) higher risk of having non-fatal and fatal cardiac events, respectively. The present study highlights the important role of depression in the context of marital relationships among ACS patients. Secondary public health care intervention programmes are needed to improve patient outcomes and minimise disease burden in clinical and community setting.